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ISSN 1454-7406

UNIVERSITATEA DE TIINE AGRICOLE


I MEDICIN VETERINAR
ION IONESCU DE LA BRAD IAI

ASOCIAIA GENERAL A MEDICILOR VETERINARI


DIN ROMNIA

LUCRRI
TIINIFICE

VOL. 49 (8)

MEDICIN VETERINAR

EDITURA ION IONESCU DE LA BRAD

IAI - 2006
Volumul a fost editat cu sprijinul financiar al
Ministerului Educaiei i Cercetrii

COLECTIVUL DE COORDONARE:
Redactor responsabil: Prof. dr. Gerard JITREANU
Redactor adjunct: Prof. dr. Vasile VNTU
Membri: Prof. dr. Neculai MUNTEANU
Prof. dr. Constantin LEONTE
Prof. dr. Gic GRDINARIU
Prof. dr. Mircea-Ioan POP
Prof. dr. Mihai CARP-CRARE

COLEGIUL DE REDACIE:
Redactor responsabil: Prof. dr. Mihai CARP-CRARE
Redactor adjunct: Conf. dr. Liviu MIRON
Membri: Prof. dr. H.C. Ioan PAUL
Prof. dr. H.C. Liviu RUNCEANU
Prof. dr. Octavian OPREAN
Prof. dr. Corneliu COTEA
Prof. dr. Tudor PERIANU
Prof. dr. Ioan COMAN
Prof. dr. Elena VELESCU

Refereni tiinifici: Prof. dr. Franois CRESPEAU


Prof. dr. Octavian OPREAN
Prof. dr. Marc ELIOT
Prof. dr. Gheorghe SOLCAN

ISSN 1454-7406

Editura Ion Ionescu de la Brad Iai


Lucrri tiinifice vol. 49 seria Medicin Veterinar

PARTICULARITI MORFOLOGICE ALE SISTEMULUI


TENDO-FASCIAL I LIGAMENTAR AL AUTOPODIULUI
TORACIC LA CERB (CERVUS ELAPHUS)
MORPHOLOGICAL PARTICULARITIES OF THE TENDO-FASCIAL AND LIGAMENTAR
SYSTEM OF THE THORACIC AUTOPODIUM IN CERVUS ELAPHUS

V. COOFAN, Valentina HRICU


E.V. INDILAR
U.S.A.M.V. Iai
A.C. HRICU
D.S.V.S.A. Iai

The research performed on 10 thoracic autopodia of Cervus elaphus


highlighted some interesting morphological particularities of the systems:
fascial, tendinous and ligamentar, not mentioned in the bibliography.
Fascia palmaris in regio metacarpophalangea is split in four layers,
from which originates: two ligamenta transversa (superficialis et
profundum), placed between the basis of the secondary fingers II and V,
two ligamenta abaxialia proximalia, which connect the secondary fingers to
the metacarpus, two ligamenta abaxialia distalia, placed between the
secondary fingers and the distal phalanx of the principal fingers and two
rami digitales palmares which inserts bifid on the falanges distales of the
principal fingers.
Tendines mm. extensores et flexores distributes only to the principal
fingers.
M. flexor digitalis superficialis presents two corpora, continued with
tendons which mentain their independence to the distal insert.
Tendo m. flexoris superficialis IV surpasses the small palmar
carpian sheath, and tendo m. flexoris superficialis III surpasses the great
carpian sheath, surrounded by a cylindric sheath, formed of m. flexor
digitalis profundus.
M. interosseus, tendinous, emits two palmar straps which, together
with the superficial flexor muscles, form long individual rings for the
branches of m. flexor digitalis profundus. Its axial and abaxial straps are
equal and unite simetrically with the tendons of mm. extensores proprii III
et IV in the plan of the proximal phalanges.
Ligamenta interdigitalia distalia are represented by two ligamenta
cruciata (palmaris et dorsalis) and a ligamentum transversum. Ligg.
cruciata are made each of two bundles which cross and reciprocal
perforate in the interdigital space.
In the thickness of the ligamentum capsularis of the regio
metacarpophalangea there are two dorsal sesamoids, with tendency of
ossification, and on the distal insert, in each tendon of m. extensor dig.
communis develops a reduced cartilaginous sesamoid

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Between the proximal phalanges, there is a strong proximal


interdigital ligament.

Key Words: anatomy, autopodium, Cervus elaphus

Studiul anatomic efectuat pe autopodiile toracale de Cervus elaphus a


evideniat aspecte morfologice inedite ale sistemului tendo-fascial i ligamentar,
nemenionate n literatura de specialitate.

MATERIAL I METOD

Cercetrile au fost efectuate pe 10 autopodii toracale de cerb prin metoda


diseciei stratigrafice. Dup piesele disecate au fost efectuate numeroase
fotografii i plane anatomice, din care 4 nsoesc textul redactat.

REZULTATE OBINUTE

Fascia carpian dorsal, mai puternic dect la taurine, particip dup


norma comun la formarea tecilor carpiene prin care trec tendoanele muchilor
extensori.
Fascia metacarpian i acropodial dorsal este degenerat n esut
conjunctiv lax, exceptnd extremitatea distal a metapodiului i regiunea
degetelor, unde formeaz teci tubulare proprii, nguste, prin care trec ramurile
tendonului muchiului extensor digital comun.
Fascia carpian palmar, clivat n dou lame, delimiteaz marea i mica
teac carpian palmar; ambele teci sunt mai nguste dect la taurine.
Fascia metacarpian palmar (e), foarte puternic, formeaz peretele
palmar al tecii n care sunt adpostite tendoanele muchilor flexori digitali i m.
interosos median. Proximal articulaiei metacarpo-falangiene se cliveaz ntr-o
lam superficial i una profund, care se continu n regiunea buletului.
n regiunea articulaiei buletului, n planul degetelor secundare, fascia
palmar se cliveaz, dispunndu-se pe patru straturi, din care se difereniaz
adevrate ligamente, particulare speciei.
Stratul 1, propriu regiunii, este reprezentat de o aglomerare de fibre elastice
ce structureaz ligamentul transvers superficial (d), dispus ntre bazele degetelor
secundare. Totodat, de pe baza fiecrui deget secundar fascia structureaz dou
ligamente abaxiale: proximal i distal.
Ligamentul proximal (g) are traiect ascendent i se inser bifid pe
metacarpul principal i pe m. interosos median; lig. distal (g) are traiect
descendent i se inser cu un fascicul scurt pe brida abaxial a m. interosos
median i cu unul lung, puternic, pe faa abaxial a falangei distale.
Stratul 2 continu lama superficial a fasciei metacarpiene i se mparte n
dou ramuri divergente (e) care coboar pe feele palmare ale degetelor
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principale. n planul falangei proximale, fiecare ramur fuzioneaz cu o ramur


similar provenit din stratul 4 (e) i, dup un scurt traiect, emite dou bride care
se inser pe faa axial i, respectiv, abaxial a falangei distale.
Stratul 3, propriu regiunii, este reprezentat de ligamentul transvers profund
(f), care leag ntre ele degetele secundare.
Stratul 4 continu lama profund a fasciei metacarpiene i se mparte n
dou ramuri care fuzioneaz cu cele provenite din stratul 2.
n plan profund, fascia acropodial palmar structureaz, la fel ca la
taurine, ligamentele inelare care contenioneaz tendoanele flexorilor digitali (a,
a, a).
Tendoanele de pe faa dorsal a meta- i acropodiului aparin muchilor
extensor digital comun i extensori digitali proprii III i IV (5, 6, 6); au
distribuie i inserii asemntoare cu cele descrise la taurine.
M. flexor digital superficial prezint dou buri musculare continuate cu
cte un tendon ce-i menine independena pn la inseria distal; n acest
context, putem vorbi de flexorul superficial al degetului III (2) i, respectiv, al
degetului IV (1).
Tendonul m. flexor superficial al degetului III parcurge marea teac
carpian palmar, unde este nconjurat de teaca cilindric (3) format de
tendonul m. flexor profund, iar tendonul m. flexor superficial al degetului IV
alunec prin mica teac carpian palmar.
n regiunea palmar a metacarpului cele dou tendoane ale flexorilor
superficiali parcurg un traiect paralel, n contact cu tendonul m. flexor profund.
Proximal articulaiei buletului, fiecare tendon primete o brid a m. interosos
median (4), formnd un inel (4) de forma unei teci lungi (cca 6 cm) prin care
trece ramura corespondent a m. flexor profund. Cele dou teci sunt unite ntre ele
printr-o lam conjunctiv. La extremitatea distal este perforat de ramura
corespondent a flexorului profund i se inser bifid pe marele burelet glenoidian.
Tendonul m. flexor digital profund (3), unic, parcurge marea teac carpian
palmar, nivel la care formeaz o teac cilindric n care este inclus tendonul m.
flexor superficial al degetului III. Proximal articulaiei buletului, tendonul m.
flexor profund se mparte n dou ramuri egale i simetrice care alunec separat
prin inelele formate de mm. flexori superficiali i bridele m. interosos median. n
regiunea acropodial traiectul i inseriile tendoanelor muchilor flexori nu
prezint diferene fa de bovine.
M. interosos median, pur tendinos, ocup toat faa palmar a metacarpului
principal (4). n treimea distal a regiunii, emite dou bride palmare care particip
la formarea inelelor pentru flexorul profund (4).
Proximal articulaiei metacarpo-falangiene, interososul median se mparte
la fel ca la taurine. Bridele axial (4IV) i abaxial (4III) pentru extensorul propriu
al degetelor sunt egale, puternice i, spre deosebire de taurine, abordeaz
extensorul simetric, n planul treimii mijlocii a falangei proximale.

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Fasciile, tendoanele i ligamentele metapodiului i acropodiului toracic


la Cervus elaphus
I - Aspectul palmar; II - Aspectul medial;

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Fasciile, tendoanele i ligamentele metapodiului i acropodiului toracic


la Cervus elaphus
III - Aspectul dorsal;

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Fasciile, tendoanele i ligamentele metapodiului i acropodiului toracic


la Cervus elaphus
IV - Aspectul axial

Legenda comun pentru fig. I, II, III i 4:


1 - Tendonul m. flexor superficial al degetului IV; 2 - Tendonul m. flexor superficial al
degetului III; 3-3 - M. flexor digital profund (3 - Tendonul comun; 3 - Teaca pentru
IV
flexorul superficial; 3 - Tendonul degetului III; 3 - Tendonul degetului IV); 4-4 - M.
interosos median (4 - Tendonul comun; 4 - Brida palmar; 4 - Inelul pentru flexorul
profund; 4III, 4IV - Bridele abaxial i axial pentru m. extensor digital propriu); 5-5 - M.
extensor digital comun (5 - Tendonul comun; 5,5 - Inseriile distale cu sesamoizii
cartilaginoi); 6 - Tendonul m. extensor propriu al degetului III; 6 - Tendonul m. extensor
propriu al degetului IV; 7 - Lig. colateral abaxial al articulaiei metacarpo-falangiene; 8,8 -
Lig. colateral abaxial al articulaiei interfalangiene proximale (8 - Fasciculul palmar 8 -
Fasciculul dorsal)

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a-c - Ligamentele interdigitale distale (a - Lig. decusat palmar; b - Lig. decusat dorsal; c -
Lig. transvers); c - Lig. interdigital proximal; c - Suprafaa de inserie a lig. interdigital
proximal; a,a,a - Ligamentele inelare; d - Lig. transvers superficial; e - Fascia
metacarpian palmar; e-e - Ramurile fasciale digitale palmare; f - Lig. transvers profund;
g - Lig. colateral abaxial proximal al degetului II; g - Lig. colateral abaxial distal al
degetului II; h - Falanga proximal a degetului III; hI - Falanga medie a degetului IV; hII -
Metacarpul III; hIII - Metacarpul IV; i - Sesamoidul dorsal; j - Marele sesamoid; l -
Sesamoidul din grosimea tendonului m. extensor digital comun; m - Micul sesamoid

Ligamentele articulaiilor acropodiale au dispoziie asemntoare cu cele


descrise la taurine. Se remarc ns, dezvoltarea deosebit, n grosime i lungime,
a ligamentelor colaterale axiale ale articulaiilor interfalangiene, tendina de
osificare a sesamoizilor dorsali (i) la nivelul articulaiei metacarpo-falangiene i
apariia a cte unui sesamoid cartilaginos, redus, n grosimea inseriei distale a
ramurilor extensorului digital comun (l).
Spre deosebire de taurine, legtura dintre degetele principale se realizeaz
prin trei ligamente interdigitale distale: decusat palmar, decusat dorsal i
transvers.
Lig. decusat palmar (a), cel mai lung, este format din dou fascicule
fibroase ce se ncrucieaz n spaiul interdigital, perforndu-se reciproc. Fiecare
fascicul leag extremitatea posterioar a feei axiale a falangei distale de
tuberculul palmar al falangei medii a degetului opus.
Lig. decusat dorsal (b), cel mai puternic, la fel ca i precedentul, este
acltuit din dou fascicule ce se ncrucieaz i se perforeaz reciproc n plan
axial. Fiecare fascicul se inser distal alturi de cel palmar, iar proximal, pe
marginea axial a falangei medii a degetului opus; la acest nivel se continu cu
ligamentul inelar falangian distal, care contenioneaz tendonul m. flexor profund.
Lig. transvers (c) este reprezentat de o band fibroas dispus ntre
marginile axiale ale micilor sesamoizi.
Lig. interdigital proximal (c) este alctuit din numeroase fibre scurte care
se inser pe treimile superioare ale feelor axiale ale falangelor proximale, unde
ocup o larg suprafa rugoas (c).
Degetele secundare sunt legate ntre ele i de formaiunile adiacente prin
ligamente derivate din fascii (d, f, g, g); articulaiile interfalangiene prezint
ligamente colaterale reduse.

CONCLUZII

1. Sistemul fascial, foarte dezvoltat la Cervus, realizeaz structuri


specializate n vederea contenionrii i glisrii tendoanelor (tecile fibroase
tubulare pentru ramurile tendonului extensor digital comun, ligamentele inelare
pentru tendoanele flexorilor digitali).
n regiunea articulaiei metacarpo-falangiene, fascia palmar se cliveaz
n patru lame, din care se difereniaz formaiuni cu un important rol n apartul

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pasiv i n amortizarea ocurilor: ligamentele transverse, dispuse ntre bazele


degetelor rudimentare, bridele fasciale pentru degetele principale i ligamentele
abaxiale, care leag baza fiecrui deget rudimentar de metacarpul principal i de
falanga distal.
2. Tendoanele muchilor extensori i flexori digitali se repartizeaz
exclusiv degetelor principale III i IV.
Burile muchiului flexor digital superficial se continu cu tendoane ce-i
pstreaz independena pn la inseria distal.
La extremitatea proximal, tendonul m. flexor digital superficial III
perforeaz tendonul flexorului profund, iar la cea distal, tendoanele flexorilor
superficiali III i IV sunt perforate de ramurile flexorului profund. Acest sistem de
perforare reciproc i succesiv a flexorilor le asigur o glisare perfect n acelai
plan, determinnd o funcionare independent a degetelor III i IV.
3. Aspectele morfologice particulare ale ligamentelor sunt corelate cu
alungirea falangelor, solicitarea deosebit a articulaiilor i a spaiului interdigital
la aceast specie.
Ligamentele colaterale axiale ale articulaiilor interfalangiene sunt foarte
lungi i groase. n grosimea ligamentului capsular al articulaiei metacarpo-
falangiene se dezvolt doi sesamoizi dorsali mai mari dect la taurine i cu
tendina de osificare, iar n tendoanele muchiului extensor digital comun, la
nivelul inseriei distale, un sesamoid dorsal redus, cartilaginos.
Unirea degetelor principale se realizeaz prin trei ligamente interdigitale
distale: decusat palmar, decusat dorsal i transvers i un ligament interdigital
proximal. Ligamentele decusate sunt formate din cte dou fascicule dispuse ntre
falanga distal i falanga medie a degetului opus; n plan axial, fasciculele se
ncrucieaz i se perforeaz reciproc, fcnd schimb de fibre.
Ligamentul interdigital proximal este format din numeroase fibre scurte,
inserate pe suprafee rugoase largi, dispuse n treimea superioar a falangelor
proximale. Dezvoltarea deosebit a acestui ligament face ca flexia i extensia
degetelor s se produc concomitent.

BIBLIOGRAFIE

1. Coofan V. i col., 1999 Anatomia animalelor domestice, Vol., I, Editura Orizonturi


Universitare, Timioara.
2. Gheie V., 1971 Anatomia animalelor domestice, Vol., I, Editura Academiei RSR,
Bucureti.
3. Nickel R. i col., 1992 Lerhbuch der Anatomie der Haustiere, Band I, Verlag Paul
Parey, Berlin und Hamburg.

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ASPECTE PRIVIND MORFOLOGIA UNOR ARTICULAII


ALE MEMBRULUI PELVIN LA STRU
(STRUTHIO CAMELLUS)
ASPECTS REGARDING THE MOPRPHOLOGY OF SOME ARTICULATIONS OF
PELVIC LIMB IN OSTRICH (STRUTHIO CAMELLUS)

C. BELU, G. PREDOI, I. DUMITRESCU,


B. GEORGESCU, Carmen BIOIU,
I. TOADER, D. PUN
U.S.A.M.V. Bucureti

As the information from specialized literature are summarily we


realized a morphological study of femurotibiopatellar articulation in
ostrich. Owing to great body weight of bird, this articulation is well-
consolidated.
There are four solid ligaments two collateral and two cruciate.
The principal movements of the femurotibiopatellar articulation as a
whole are flexion and extension.

Key words: morphology, articulations, struthio.

Cu toate c literatura de specialitate cuprinde un numr nsemnat de lucrri


referitoare la biologia, tehnologia de cretere i patologia speciilor din ordinul
Ratite (1, 2, 3, 4), datele referitoare la morfologia acestor specii sunt relativ
sumare sau, de cele mai multe ori tratate subiectiv (5, 6, 7).
Lucrarea de fa i propune descrierea detaliat a unor formaiuni integrate
sistemului articular al membrului pelvin la struul african, n scopul copletrii
datelor din literatur, astfel nct aspectele descrise s poat fi de folos
clinicienilor n cazul abordrii unor conduite trerapeutice adecvate.

MATERIAL I METOD

Materialul de studiu a fost reprezentat de 5 exemplare adulte, sacrificate de


necesitate, provenite de la diferii cresctori particulari. Metoda de lucru a
constituit-o disecia, pn la limita vizibilitii folosind stereomicroscopul SMZ 2-T
Nikon. Descrierea i omologarea formaiunilor s-a realizat n corelaie cu Nomina
Anatomica Avium 1979.

REZULTATE I DISCUII

Articulaia femuro-tibio-patelar este reprezentat de un complex articular


realizat ntre oasele acestei regiuni. O prim articulaie se realizeaz ntre

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trochleea anterioar a epifizei distale a femurului i faa articulart a patelei, care,


pentru a corespunde trochleei femurale este completat cu dou mase
fibrocartilaginoase. O a doua articulaie se realizeaz ntre cei doi condili femurali
(dintre care cel medial apare mai dezvoltat) i tibie iar cea de-a treia, ntre
trochleea femural i extremitatea proximal a fibulei.

Fig. 1. Articulaia femuro-patelo-


tibial la struul african (vedere
cranial)
1. menisc medial,
2. menisc lateral,
3. ligament meniscofemural cranial,
4. ligament transvers al genunchiului,
5. ligament meniscotibial cranial,
6. ligament menisco-colateral,
7. ligament meniscofibular cranial,
8. patela.

Fig. 2. Articulaia femuro-patelo-


tibial la struul african (vedere
caudal)
1. ligament colateral medial,
2. menisc medial,
3. menisc lateral.

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ntre suprafeele articulare enunate se gsesc dou meniscuri fibro-


cartilaginoase.
Meniscul medial are o form semilunar. n partea cranial este consolidat
printr-un grup de fibre de partea medio-rostral a meniscului lateral, grup ce
reprezint ligamentul transvers al genunchiului (lig. transversum genus). Tot din
partea cranial a sa se desprinde un grup de fibre - ligamentul menisco-femural
cranial, ce fixeaz meniscul la partea lateral a amprentei ligamentului ncruciat
cranial (absent la psrile de curte). Caudal acest menisc prezint dou inserii.
Cea mai puternic este reprezentat de ligamentul menisco-tibial (lig.
meniscotibiale caudale), care nconjoar pe partea caudo-medial ligamentul
ncruciat caudal i se inser pe partea posterioar a eminenei intercondiliene a
tibiei. Cel de-al doilea ligament (lig. meniscofemorale) este mult mai redus i se
inser n fosa intercondilian, la baza condilului femural medial.
Meniscul lateral este o formaiune fibrocartilaginoas interpus ntre femur
i condilul lateral al tibiei. Are aspect trapezoidal, cu baza mare orientat cranial.
Inseria cranial a acestui menisc este reprezentat de un ligament (lig.
meniscotibiale craniale) ce se fixeaz n partea cranial a condilului tibial medial.
De asemenea, meniscul lateral este fixat pe partea cranial a extremitii
proximale a fibulei printr-un veritabil ligament meniscofibular cranial. Un al
treilea ligament foarte fin (lig. meniscocollaterale) fixeaz meniscul la ligamentul
colateral lateral al articulaiei femuro-tibiale.

Fig. 3. Articulaia femuro-


patelo-tibial la struul
african (vedere medial)
1. muschiul tibial cranial,
2. patela,
3. menisc medial,
4. ligament colateral medial,
5. menisc lateral,
6. epicondil medial.

Complexul articular este consolidat de urmtoarele ligamente:


- ligamentul ncruciat anterior (lig. cruciatum craniale) care se inser pe
vrful eminenei intercondiliene a tibiei, apoi se orienteaz caudo-dorsal, trecnd
printre meniscuri, apoi prin anul dintre cei doi condili, ajungnd n cele din
urm pe faa caudal a femurului, medial fa de fosa poplitee;

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- ligamentul ncruciat caudal (lig. cruciatum caudale) este mai puternic


dect precedentul. El se prinde caudal n partea posterioar a condilului tibial
medial, apoi trece cranio-dorso-lateral pe sub meniscul medial, pentru a se insera
cranial n anul intercondilar, mpreun cu ligamentul menisco-femural cranial;
- ligamentul colateral lateral (lig. collaterale laterale) se prinde pe
epicondilul femural lateral (slab reprezentat) i pe faa lateral a extremitii
proximale a fibulei;
- ligamentul colateral medial (lig. collaterale mediale) este inserat pe
epicondilul medial al femurului (foarte redus la stru) i pe faa medial a
extremitii proximale a tibiei.
Articulaia tibiopatelar se realizeaz ntre patel i extremitatea proximal
a tibiei. Ea este consolidat de un singur ligament ligamentul patelar (lig.
patellae) care se ntinde de la marginea ventral a patelei la creasta patelar a
tibio-tarsului. Ligamentul patelar formeaz peretele cranial al cavitii articulare a
articulaiei femuro-tibiale.

CONCLUZII

Articulaia femuro-tibial include dou meniscuri diferite ca form, bine


consolidate att la extremitatea proximal a tibiei, ct i la extremitatea distal a
femurului.
Ligamentul ncruciat caudal este mult mai bine reprezentat dect
ligamentul ncruciat cranial.
Ligamentele colaterale sunt extrem de bine reprezentate, fapt care
contribuie la mrirea preciziei micrilor (articulaia realizeaz micri de
extensie-flexie).
BIBLIOGRAFIE
1. Beattie I., 1992 - Applied ostrich physiology and anatomy. Ostrich workshop for
veterinarians; April 11-12, Zimbabwe. Portland, Oregon: Island Ostrich Ranch;
[1992]10 p.
2. Bezuidenhout A., Burger W., 1993 - The Incidence of Tibiotarsal Rotation in the
Ostrich (Struthio-Camelus). Journal of the South African Veterinary Association;
64(4): 159-161.
3. Fedak M., Seeherman H., 1979 - Reappraisal of energetics of locomotion shows
identical cost in bipeds and quadrupeds including ostrich and horse. Nature
1979; 282: 713-716.
4. Fowler M.E., 1992 - Clinical anatomy of ratites. Proceedings of the Annual
Conference of the Association of Avian Veterinarians; September 1-5, 1992; New
Orleans. Florida: Association of Avian Veterinarians; 1992 307-309.
5. Pavaux C., Lignereux Y. - A Myological Dissection of the Shank and Foot in an
Ostrich (Struthio-Camelus). Journal of Veterinary Medicine Series C 1995; 24(2):
127-131.
6. Ionescu V. i col., 2003 Struul cretere i patologie, Ed. CORAL SANIVET,
Bucureti.
7. Ioni Mariana, Mitrea L., 2006 Biologie animal, Ed. Medical veterinar,
Bucureti.

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ASPECTE HISTOLOGICE LA NIVELUL REELELOR


ADMIRABILE INTRACRANIENE LA MAMIFERELE
DOMESTICE
THE HISTOLOGICAL ASPECTS AT THE LEVEL OF RETE MIRABLE
AT DOMESTIC MAMMALS

I. DUMITRESCU, G. PREDOI,
N. CORNIL, C. BELU, B. GEORGESCU,
Florina DUMITRESCU, Petronela ROU
U.S.A.M.V. Bucureti

The study of histological aspects was made by using the rete


mirables from two taurines and four ovines.
The space which is circumscribed by the venous cavernous sinus and
without possibility to distend it, which is offered to the rete mirable induces
an important polymorphism to the blood sanguiferous vessels which
structures the rete mirable.
The small arterys vascular wall from the rete has the median tunic
flanked on both sides by endotelium, the adventitia beeing absent.
The arterial endotelium can structure small intravascular pillows
with meaning to control the sanguin flow capacity at the level of
encephalon.

Key words: histological, rete mirables, mammals

Reelele admirabile epidurale joac, la speciile beneficiare, un rol


fundamental n reglarea debitului sanguin la nivelul encefalului (1, 6, 9). ntruct
n literatura de specialitate consultat nu am ntlnit date cu privire la
particularitile histologice ale arteriolelor ce structureaz reeaua admirabil
epidural, am considerat necesar studierea histologic a acestor structuri (2, 3, 4,
5, 7, 8, 10, 11, 12).

MATERIAL I METOD

Studiul s-a realizat folosind reele admirabile epidurale provenite de la dou


tauriune i patru ovine. n acest scop, imediat dup sacrificarea animalelor am
realizat splarea substratului vascular de la nivelul reelelor admirabile epidurale,
injectnd pe artera carotid comun ser fiziologic, pn cnd pe artera simetric
soluia de ser fiziologic exprimat nu mai prezint urme de snge. Ulterior, am
procedat la recoltarea reelei admirabile epidurale ptrunznd prin regiunea
subsfenoidal. Dup recoltare, am imersat reeaua n soluie de formaldehid
10%, n vederea realizrii ulterioare de preparate histologice. Studiul histologic s-
a fcut n cadrul laboratorului de histologie al Facultii de Medicin Veterinar

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Bucureti. Imaginile obinute au fost studiate i fotografiate cu ajutorul


microscopului optic Nikon, prevzut cu dispozitiv de microfotografiere. Metoda de
coloraie a fost cea cu hematoxilin - eozin, Giemsa i cea cu orcein pentru
evidenierea fibrelor elastice din peretele vascular.

REZULTATE I DISCUII

Spaiul limitat, i fr posibilitate de dilatare de la nivelul sinusului venos


cavernos, impune o mare variabilitate conformaional a vaselor ce compun
reeaua admirabil epidural. Aceast variabilitate este evideniat i histologic
prin polimorfismul vaselor de la nivelul reelei.
Reeaua admirabil epidural este strbtut de filete nervoase cu origine n
principal n ortosimpaticul suprasfenoidal dar i ramificaii din perechile III, IV i
VI de nervi cranieni. Histologic aceste fibre nervoase apar separate prin septumuri
conjunctive de peretele vascular.
n urma studierii preparatelor histologice efectuate s-a observat c vasele
mari precum trunchiul bazilar, colateralele acestuia, artera carotid intern
reconstituit din reeaua admirabil epidural, precum i terminalele acesteia etc.
prezint caracteristicile arterelor de tip muscular.
n structura peretelui predomin tunica medie, compus n principal din
leiocite, dispuse n mai multe straturi de celule concentrice. Printre celulele
musculare se ntlnesc fibre elastice identificabile prin coloraie cu orcein i de
colagen.
Adventicea cuprinde multe fibre elastice i de colagen, fibroblaste, vasa
vasorum, limfocite i nervi vasorum. Arterele de tip muscular primesc o bogat
inervaie vegetativ, ce le permite s desfoare o intens i rapid vasomotricitate
(vasoconstricie i vasodilataie).
Majoritatea arterelor ce intr n alctuirea reelelor admirabile epidurale
sunt ns arteriole cu lumenul mult mai redus dect lumenul arterelor musculare
mici, dar care prezint un perete gros n raport cu diametrul lumenului.
Delimitarea ntre tunici apare mai puin evident iar diametrul este sub 0.5 mm.
De cele mai multe ori deosebirea dintre arterele musculare mici i arteriole apare
dificil de sesizat.
n general, reelele admirabile includ arteriole mari, caracterizate prin
urmtoarele: intima, foarte subire, format din endoteliu, strat subendotelial cu
fibre de colagen i elastice; media, format din 2-6 straturi concentrice de celule
musculare netede, fine fibre elastice i de colagen, n cazul reelei admirabile
epidurale, peretele fiind limitat la exterior chiar de endoteliul sinusului venos ce
nclude reeaua. Adventicea este absent n aceste cazuri.
La nivelul arteriolelor amintite, s-a observat ngrori ale intimei, ce
proemin n lumen i l nchide atunci cnd arteriola sufer un proces de
vasoconstricie. Prezena structurilor vasculare speciale de tipul pernuelor
arteriale este dictat de necesitatea asigurrii unui debit sanguin variabil, corelat

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

cu necesarul sanguin al encefalului la un anumit moment dat. Aceleai structuri se


ntlnesc i n organe parenchimatoase cu activitate intens (rinichi, gland
tiroid).

Foto 1 Aspect de ansamblu al reelei admirabile,


col. Giemsa, ob. 4 x,
1 - ramificaii arteriale; 2- spaii venoase ale sinusului cavernos; 3 - endoteliu arterial;
4- endoteliul venos; 5 - leiocite.

Foto 2 Pernue intravasculare la nivelul unor vase din reeaua admirabil


col. H-E, ob. 10 x,
1 - vas; 2- pernue intravasculare.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Foto 3 Fibre elastice n pereii vaselor reelei,


col. orcein; ob. 10 x,
1 - lumen; 2- perete arterial, 3 - spaiul sinusului cavernos.

CONCLUZII

1. Spaiul limitat i fr posibilitate de dilatare oferit reelelor admirabile


epidurale de ctre sinusul venos cavernos, impune un polimorfism accentuat al
vaselor ce structureaz reeaua admirabil.
2. Reeaua admirabil epidural este strbtut de filete nervoase cu origine
n principal n ortosimpaticul suprasfenoidal dar i ramificaii din perechile III, IV
i VI de nervi cranieni. Histologic aceste fibre nervoase apar separate prin
septumuri conjunctive de peretele vascular.
3. Tunica medie a peretelui vascular este structurat din fibre musculare
netede dispuse n straturi concentrice; la acest nivel sunt abundente i fibrele
elastice evideniate prin coloraia cu orcein.
4. Media arteriolelor ce structureaz reeaua admirabil epidural este
tapetat la interior de endoteliul arterial, iar la exterior de ctre endoteliul
sinusului venos cavernos.
5. La nivelul arteriolelor din reea, se ntlnesc ngrori ale intimei ce
nchid lumenul atunci cnd arteriola sufer un proces de vasoconstricie. Aceste
structuri endoteliale numite pernue vasculare particip la asigurarea unui debit
sanguin variabil la nivelul encefalului.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

BIBLIOGRAFIE

1. Baldwin B.A., 1964 The anatomy of the arterial supply to the cranial regions of the
sheep and ox. Am. J. Anat., 115, 101 118.
2. Barone R., 1996 - Anatomie compare de mamiferes domestiques, Tome V
Angiologie, Ed. Vigot, Paris.
3. Botrel S., Cotea C., Gaboreanu M., 1982 Histologie i embriologie, Ed. Didactic
i Pedagogic, Bucureti.
4. Coman T., Cornil N., 1999 Embriologie veterinar. Ed. Fundaiei Romnia de
mine, Bucureti, 1999.
5. Cornil N., 1993 Curs de biologie celular, histologie, embriologie, vol. 3. Lito.
AMD UAMV, Bucureti.
6. De Lahunta A., Habel E.R., 1986 Applied Veterinary Anatomy, W.B. Saunders
comp..
7. Diculescu I., Doina Onicescu, 1987 Histologie medical, vol. I II. Ed. Medical,
Bucureti.
8. Godinho H.P., 1968 A comparative anatomical study of the cranial nerves in goat,
sheep and bovine; their distribution and releated autonomic components. Ph. D.
Thesis, Iowa State University Amer.
9. Lagutchick M.S., Sturgis J.W., Martin D.G., Bley J.A., 1992 Review of the
carotid artery loop procedure in sheep. Journal of Investigative Surgery 5 (2), 79
89.
10. Patea E., Cornil N., 1973 Morfotopografia lojei hipofizare la oaie. n Lucr. t.
IANB, CXVI, Bucureti, 7 11.
11. Predoi G., Belu C., 1994 Cercetri comparative privind vascularizaia venoas
cefalic la ovine i caprine. Conferina Jubiliar 20 de ani de nvmnt
superior medical veterinar n Republica Moldova, Chiinu.
12. Sisson S., Grossman J.D., Getty R., 1975 - The Anatomy of the Domestic Animals,
vol. I. W.B. Saunders Comp., Philadelphia London Toronto

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Universitatea de tiine Agricole i Medicin Veterinar Iai

STUDII PRIVIND MORFOSTRUCTURA BURSEI


CLOACALE LA PALMIPEDELE DOMESTICE
STUDIES CONCERNING MORPHOLOGY OF CLOACAL BURSA
IN DOMESTIC PALMIPEDES

B. GEORGESCU, G. PREDOI, N. CORNIL,


Emilia CIOBOTARU, C. BELU,
L. ENACHE, I. DUMITRESCU, A.I. TOADER
U.S.A.M.V. Bucureti

Cloacal bursa is a primary specific lymphoid organ of birds. This


organ has a median dorsal position given cloaca. Cloacal bursa is a sack-
shaped organ, with cranially orientated free extremity.
The study was performed on 30 individuals of palmiped domestic
species: domestic duck (Anas domesticus), Muscovy duck (Cairina
moschata) and domestic goose (Anser domesticus). 10 individuals of 6
mouth-old were studied for each species. Cloacal bursa is fusiform organ in
all studied species.
The values of lengh and diameter were between 2.5 - 3.5 cm,
respectively 1.5 - 1.7 cm in ducks. The lengh and diameter of cloacal bursa
in goose were between 2.7 3.5 cm, respectively 1.5 2.0 cm.
Histologically, submucosal infiltration of lymphociytes occured in
dorsal cloacal wall in goose. Tending of lymphonodules formation was
obvious.

Key words: morphology, cloacal, palmipedes

Bursa cloacal este un organ limfoid primar caracteristic psrilor. Aceast


formaiune se afl dispus n plan median dorsal fa de cloac. Bursa cloacal are
aspectul unui sac cu extremitatea liber orientat cranial.

MATERIAL I METOD

Studiul a fost realizat pe 30 de exemplare aparinnd celor 3 specii de


palmipede domestice si anume raa domestic (Anas domesticus), raa leeasc
(Cairina moschata) i gsca domestic (Anser domesticus), pentru fiecare specie
studiindu-se cte 10 exemplare n vrsta de 6 luni.
Dup sacrificarea psrii prin injecie intrabulbar de sol. Xilina 1% i
deschiderea cavitii abdominale se ridic ansele intestinale i se abordeaz
segmentul intestinal terminal. Se identific cloaca i pe peretele dorsal al acesteia
se remarc prezena bursei cloacale. Se secioneaz peretele cloacal anterior i
posterior de pediculul bursal i se recolteaz bursa mpreun cu o redus
poriune de perete cloacal n vederea studiului histologic.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Pentru studiul histologic bursele cloacale rezultate au fost incluse n


parafin si apoi realizate preparate ntre lam si lamel, preparate colorate cu
metode uzuale (hematoxilin, eozin, albastru de metil).
REZULTATE I DISCUII

Forma bursei cloacale nu difer n cadrul speciilor de psri studiate, la


toate speciile de palmipede bursa cloacal aprnd fusiform (fig. 1, 2)

Fig. 1 - Aspectul bursei


cloacale (1) la ra leeasc
n vrst de 6 luni

Fig. 2 - Aspectul bursei


cloacale (1) la gsc n
vrst de 6 luni

Dimensiunile bursei cloacale difer n funcie de specie. La psrile la care


bursa se afl n deplin activitate (aproximativ vrsta de 6 luni) au fost constatate
urmtoarele dimensiuni: la speciile de rae au fost obinute valori medii de 2,5
3,5 cm lungime i 1,5 1,7 cm diametru; iar la gsc lungimea medie a fost de
2,7 3,5 cm i diametrul de 1,5 2,0 cm. Valorile medii ale bursei cloacale sunt
prezentate n tabelul 1.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Tabelul 1
Dimensiunile medii ale bursei cloacale la unele specii de psri domestice

Vrsta Dimensiuni medii (cm)


Specia
(luni) Lungime Diametru
Rae
6 2,5 3,0 1,5 1,7
(Anas domesticus + Cairina moschata)
Gsc
6 2,7 3,5 1,5 2,0
(Anser domesticus)

La toate speciile de psri bursa este un organ cavitar ataat peretelui dorsal
al cloacei printr-un scurt pedicul. Lumenul bursal comunic printr-un canal (canal
bursal) dispus n pedicul cu cavitatea proctodeumului.
Cu toate c literatura consultat menioneaz prezena destul de frecvent a
unor formaiuni bursale accesorii, la psrile studiate aceste formaiuni nu au fost
observate.
n general histostructura bursei cloacale la psrile studiate respect
structura histologic prezentat n lucrrile consultate. La toate speciile de psri
peretele bursei este format din trei tunici suprapuse: seroas, musculoas i
mucoas.
Seroasa este reprezentat de peritoneul local care acoper bursa.
Musculoasa este format din fibre musculare netede dispuse pe dou straturi:
longitudinal extern i circular intern.

Fig. 3 - Burs cloacal la


ra n vrst de 6 luni
(H.E.A., ob. 4).
Foliole foarte lungi ce
prezint la vrf numeroase
celule productoare de
mucus

Mucoasa ocup cea mai mare parte din grosimea peretelui bursei avnd
lamina propria ocupat aproape n totalitate de foliculi limfoizi foliculi bursali
(foliculi Stannius). Mucoasa este puternic cutat, constituindu-se pliuri groase
(foliole bursale) (fig. 3, 4), al cror centru este ocupat de foliculi, cu contur
poliedric, iar suprafaa plurilor este acoperit cu un epiteliu simplu sau
pseudoprismatic columnar.

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Fig. 4 - Burs cloacal la


ra de 6 luni (H.E.A., ob.
20). Se remarc aspectul
clar al celulelor epiteliale de
la nivelul vrfului foliolei
bursale

La nivelul epiteliului sunt prezente trei tipuri de celule columnare (tipul I, II


i III) n funcie de granulele pe care le conin. Caracterul de epiteliu simplu
columnar este mai evident atunci cnd acesta acoper proeminenele determinate
de proliferarea fiecrui folicul. Epiteliul prolifereaz n profunzime, celulele
epiteliale amestecndu-se cu celulele din foliculi.
Foliculii bursali pot prezenta dou aspecte: aspect uniform (n cazul
foliculilor primari) i foliculi cu dou zone, cortical i medular (n cazul
foliculilor secundari) (fig. 5, 6, 7).
Foliculii bursali secundari prezint la periferia medularei un strat de celule
epiteliale nedifereniate, cubice i un strat de capilare. Corticala este separat de
medular printr-o membran bazal. Zona cortical apare mai nchis la culoare i
cuprinde numeroase limfocite, limfoblaste, celule n mitoz i numeroase
macrofage. Medulara apare mai deschis la culoare i cuprinde celule
nedifereniate, limfoblaste i limfocite.

Fig. 5 - Burs cloacal la


ra de 6 luni (H.E.A., ob.
20). Foliculii primari ncep
s se tranforme n foliculi
secundari.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Fig. 6 - Bursa cloacal la


gsc n vrst de 6 luni
(H.E.A., ob. 10). Se observ
c epiteliul asociat
foliculului i limitanta
foliculului se afl n
continuitate
cu membrana bazal a
epiteliului bursal

Fig. 7 - Burs cloacal la


gsc n vrst de 6 luni
(H.E.A., ob. 40). Folicul
bursal epiteliul asociat
foliculului i limitanta n
continuitate direct cu
membrana bazal a
epiteliului

Fig. 8. - Perete dorsal al


cloacei la gsc n vrst
de 3 luni (H.E.A., ob. 20).
Noduli limfoizi n curs de
organizare

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esutul limfoid este prezent i n peretele dorsal al cloacei. S-a observat


infiltrarea cu limfocite a submucoasei i tendina de a ajunge pn n vecintatea
musculoasei. De asemenea s-a remarcat tendina de organizare a limfocitelor n
limfonoduli. Epiteliul, de tip digestiv, acoper agregatele limfoide i include un
mare numr de celule caliciforme. Acest esut limfoid n peretele dorsal al cloacei
este ns inconstant. n cazul psrilor studiate a fost observat numai la 3 gte n
vrst de 6 luni (fig. 8).

CONCLUZII

Pe baza rezultatelor cercetrilor proprii efectuate pe 90 psri domestice


aparinnd unui numr de 3 specii de palmipede domestice ( ra, ra leeasc,
gsc) se pot desprinde urmtoarele concluzii cu privire la morfostructura bursei
cloacale:
1. Bursa cloacal reprezint un organ limfoid primar caracteristic psrilor.
Este dispus dorsal, pe peretele proctodeumului i cavitatea sa comunic cu
cavitatea proctodeal prin canalul bursal situat n pedicul.
2. Forma i dimensiunile bursei difer de vrst i de specie. La palmipede
forma este alungit, ovoidal i dezvoltarea maxim este atins la vrsta de 6 luni.
3. Din punct de vedere histologic peretele bursal apare alctuit din trei
tunici suprapuse (seroas, musculoas i mucoas). Seroasa este reprezentat de
peritoneul local. Musculoasa este format dintr-un strat longitudinal extern i un
strat circular intern.
4. Lamina proprie a mucoasei la toate speciile este ocupat aproape n
totalitate de foliculi limfoizi primari i secundari. Foliculii bursali secundari
prezint dou zone: medular i cortical. Medulara apare mai deschis la culoare
i cuprinde celule nedifereniate, limfoblaste i limfocite. Corticala, mai nchis
colorat, cuprinde numeroase limfocite, limfoblaste, celule n mitoz i
numeroase macrofage.
5. La gsc a fost observat infiltrarea cu limfocite a submucoasei peretelui
dorsal al cloacei. S-a remarcat tendina de organizare a limfocitelor n
limfonoduli.

BIBLIOGRAFIE

1. Breuil G.A., 1980 Aspects normaux et pathologiques du sytme lymphode des


oiseaux. Thse, Ecole Nationale Vtrinaire de Toulouse.
2. Coofan V., Palicica R., Valentina Hricu, Carmen Gan, Enciu V., 2000
Anatomia animalelor domestice. Vol. III. Ed. Orizonturi Universitare, Timioara.
3. Georgescu B., 2001 Cercetri morfotopografice asupra formaiunilor limfoide la
psrile domestice. Tez, F.M.V. Bucureti.
4. King A.S., J. McLelland, 1984 BIRDS their structure and function. Second
Edition. Ed. Bailliere Tindall, London Philadelphia Toronto Mexico City
Rio de Janeiro Sydney - Tokyo Hong Kong.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

5. Nickel R., Schummer A., Seiferle E., 1973 Lehrbuch der Anatomie der Haustiere.
Band V Anatomie der Hausvgel. Verlag Paul Parey, Berlin und Hamburg.
6. Patea E., Coofan V., Chiescu t., Miclea M., Cornil N., Nicolescu V., Radu C.,
Popovici I., Palicica R., 1985 Anatomia comparat a animalelor domestice.
Vol. I II. Ed. didactic i pedagogic, Bucureti.
7. Sisson S., J.D. Grossman, R. Getty, 1975 The Anatomy of the Domestic Animals.
Vol. I II (Fifth Edition). W.B. Saunders Company, Philadelphia London
Toronto.
8. * * * - Nomina Anatomica Avium. Academic Press, London Toronto New York
Sydney San Francisco, 1979.
9. * * * - Nomina Anatomica Veterinaria (Fourth Edition), together with Nomina
Histologica (Revised Second Edition) and Nomina Embryologica Veterinaria.
Zrich and Ithaca, New York, 1994.

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CERCETRI PRIVIND VASCULARIZAIA VENOAS A


GLANDEI MAMARE LA OAIE
RESEARCH REGARDING THE VENOUS VASCULARITY OF MAMMARY GLAND
IN EWE

G. PREDOI, C. BELU, I. DUMITRESCU,


B. GEORGESCU, N. CORNIL,
Carmen BIOIU, I. TOADER
U.S.A.M.V. Bucureti

The study material was represented by 15 sheeps, from which were


collected udders. We followed the detailed description of the venous
formations from the mammary gland and also the description of any
individual particularities which hadnt been discovered until recently days
in the literature.
The drainage venous ways and its many anastomosis between the
halfs of the udder were represented.

Key words: vascularity, mammary gland, sheep

Literatura de specialitate cuprinde relativ puine date referitoare la


vascularizaia glandei mamare la ovine, descrierea formaiunilor vasculare fiind
relativ sumar, uneori fiind ntlnite date contradictorii.
Din acest motiv am considerat necesar continuarea i aprofundarea
cercetrilor n scopul completrii datelor existente i pentru clarificarea datelor
contradictorii.

MATERIAL I METOD

Materialul de studiu a fost reprezentat de 15 exemplare de ovine,


caracterizate printr-o heterogenitate accentuat a vrstei, rasei, strii de
ntreinere i a strii fiziologice. Unele animale au fost sacrificate n abator sau n
diferite puncte de tiere iar altele au fost utilizate n scop didactic n cadrul
laboratorului de Anatomie de la F.M.V. Bucureti.
Formaiunile venoase au fost cercetate dup injectarea lor cu o substan
plastic contrastant (AGO), dizolvat n aceton (20-30%). Injectarea masei
plastice dizolvate s-a realizat dup splarea patului venos cu ser fiziologic, prin
vena pudend extern. Dup injectare piesa a fost supus unui proces de
formolizare timp de 24 de ore.
Studierea, descrierea i omologarea formaiunilor s-a fcut n concordan
cu Nomina Anatomica Veterinaria - 1994.

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REZULTATE I DISCUII

Venele principale de descrcare ale ugerului sunt reprezentate prin cele


dou vene pudende externe. Secundar sngele este descrcat i prin venele
pudende interne (Foto 1).

Foto 1 - Aspect de ansamblu al venelor glandei mamare la ovine - vedere dorsal


(original)
1- vena pudend extern; 2- vena pudend intern; 3 - rdcina lateral a venei
pudende externe; 4- rdcina median; 5- rdcina medial; 6- limfonod mamar

Fiecare ven pudend extern rezult prin confluarea la nivelul inelului


inghinal superficial a 3 rdcini venoase: lateral, median, medial.
Rdcina lateral are calibrul cel mai redus i urmrete marginea lateral a
ugerului, avndu-i originea la extremitatea antero-lateral a acestuia.
Rdcina median urmrete aproximativ planul dorso-median al fiecrei
mamele, fiind cea mai dezvoltat dintre rdcinile venei pudende externe
(Foto 2-4). Originea acestei ramuri o constituie n regiunea abdominal ventral,
anterior de baza ugerului, vena epigastric caudal superficial. n a doua
jumtate a gestaiei i n perioada lactaiei, vena epigastric caudal superficial
apare foarte dezvoltat i reprezint principalul vas de descrcare a sngelui venos
de la nivelul ugerului.
n general, rdcinile mediane au calibru inegal (80% din cazuri). Cranial
de baza ugerului, la aproximativ 3-4 cm, n 30% din cazuri, rdcinile mediane
ale celor 2 vene pudende externe sunt conectate transversal printr-o ramur cu

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

traiect oblic i uor sinuoas (Foto 2-6). Pe traiect rdcina median primete
numeroi aflueni ce abordeaz trunchiul principal att din sens lateral i medial
ct i din plan profund.

Foto 2 - Aspect diferenial privind


originea i traiectul rdcinilor celor
dou vene pudende externe la oaie
(original)
1 -vena pudend extern stng;
2 - vena pudend extern dreapt;
3 - rdcina lateral;
4 - rdcina median;
5 - rdcina medial;
6 - ramur de anastomoz ntre
rdcinile mediane;
7 limfonod mamar;

La limita dintre treimea anterioar i mijlocie a bazei ugerului, rdcina


median a pudendei externe primete un puternic afluent, ce i are rdcinile att
n regiunea subcapsular n planul median ventral al ugerului, ct i la faa
medial a mamelei de partea opus. Pe tot traiectul su exist ramuri comunicante
ntre afluenii celor 3 rdcini ale venelor pudende externe. La aproximativ 2 cm
de locul de convergen a celor 3 rdcini, rdcina median a venei pudende
externe primete un ultim mare afluent, ce i are originea la faa ventral a
glandei mamare corespunztoare, n esutul conjunctiv subcutanat, apoi urc i
abordeaz rdcina la faa ventral a acesteia.
n general, rdcinile mediale au un traiect diferit pe dreapta fa de
situaia de pe partea stng. Astfel, mai frecvent (70% din cazuri) am ntlnit
situaia n care rdcina medial a venei pudende externe drepte i are originea la
extremitatea anterioar a mamelei corespunztoare, apoi n traiectul su caudal i
mrete progresiv volumul, plasndu-se pe toat lungimea la aproximativ 1,5 cm
de planul sagital al ugerului.

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Pe partea opus, rdcina medial a venei pudende externe stngi i are


originea, n aceste cazuri, n regiunea subcutanat a planului median ventral al
ugerului, apoi urc pe direcia septumului mamar median i se unete n final la
zona de convergen dintre vena pudend extern i vena pudend intern.
n 70% din cazuri rdcinile mediale ale celor dou vene pudende externe
sunt conectate n poriunea terminal printr-o ramur venoas dispus transversal
(Foto 3)

Foto 3 - Aspectul ramurii de anastomoz a rdcinilor mediale a venelor pudende


externe (original)
1 - rdcini mediale ale venelor pudende externe; 2- ramura anastomotic;
3 - rdcinile mediane; 4 - limfonodurile mamare.

Venele pudende interne se detaeaz din zona de convergen a radciniilor


median i medial ale venelor pudende externe .
n 20% din cazuri s-a observat c vena pudend intern stng apare ca o
continuare direct, n sens caudal, a rdcinii mediale aparinnd venei pudende
externe stngi.
Cele dou vene pudende interne se orienteaz caudal, n regiunea perineal
i, n 90% din cazuri, sunt anastomozate antero-ventral de arcada ischiatic printr-
o ramur dispus transversal (Foto 4).
n partea caudal a bazei ugerului, rdcinile mediale ale celor dou vene
pudende externe sunt anastomozate printr-o ramur transversal bifurcat la
extremitatea dreapt.

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Foto 4 - Aspectul venelor pudende interne atunci cnd ele sunt conectate
printr-o ramur anastomotic (original)
1- venele pudenda externe; 2- venele pudende interne; 3- ramur anastomotic;
4 - rdcina lateral a venei pudende externe; 5 - rdcina median a venei pudende
externe; 6-rdcina medial a venei pudende externe; 7- ramur anastomotic;
8 - limfocentrul mamar.

CONCLUZII

Fiecare ven pudend extern, ce reprezint vasul principal de descrcare a


ugerului, este format din trei rdcini: lateral median i medial. n a dou
parte a perioade de gestaie principalul vas de descrcare este reprezentat de vena
epigastric caudal superficial.
Cranial de baza ugerului, la aproximativ 3-4 cm, n 30% din cazuri,
rdcinile mediane ale celor dou vene pudende externe sunt conectate transversal
printr-o ramur cu traiect oblic i uor sinuoas.
Ramuri mai reduse, inconstante ca numr i topografie, conecteaz afluenii
celor trei rdcini ale venelor pudende externe.
S-au observat conexiuni transversale ntre rdcinile mediale dar i ntre
venele pudende externe.

BIBLIOGRAFIE

1. Barone, R., 1996 - Anatomie compare de mamiferes domestiques, Tome V


Angiologie, Ed. Vigot, Paris.
2. Benzie, D.; Cresswell, E.; 1962 Studies on the lactation of sheep. Res. Vet. Sci.
3(3), 231 235.

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3. Coofan, V., 1986 Anatomia i fiziologia animalelor domestice. Ed. Ceres,


Bucureti.
4. May, N.D.S., 1970 The anatomy of the sheep (a dissection manual), Third ed.
University of Queensland Press, St. Lucia, 225 240.
5. Nickel, R.; Schummer, A.; Seiferle, E., 1973 Lehrbuch der Anatomie der
Haustiere. Verlag Paul Parey, Berlin Hamburg.
6. Patea, E.; Coofan, V.; Chiescu, t.; Miclea, M.; Cornil, N.; Nicolescu, V.;
Radu, C.; Popovici, I.; Palicica, R., 1985 Anatomia comparat a animalelor
domestice, vol. I i II. Ed. didactic i pedagogic, Bucureti.
7. Sisson, S.; Grossman, J.D.; Getty, R., 1975- The Anatomy of the Domestic
Animals, vol. I. W.B. Saunders Comp., Philadelphia London Toronto.

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ANATOMICAL PARTICULARITIES OF THE PELVIS


OF THE CARPATHIAN BEAR
PARTICULARITILE ANATOMICE ALE PELVISULUI LA URSUL CARPATIN

C. SPTARU, Mihaela-Claudia SPTARU,


D. ANIA
U.S.A.M.V. Iai

At the bear similar to domestic mammals, the pelvis is composed of


(ilium, ischium and pubis), coxal articulated dorsally with the sacrum. The
ilium is particular through an iliac palette of triangular form, provided with
a wide gluteus fosse. The neck of the ilium is short and the tubercle of the
small psoas muscle if found in the vicinity of the cotillion cavity. The
ischium has a massive body, provided with a rounded off ischiatic spine.
The ischial tuberose is very developed in a triangular form. The caudal
edge of the ischium attains an ischial arch in a V shape with a dorsal
opening. The pubis has a triangular form with a sharp cranial edge and
medial of the cotillion cavity it is provided with a developed tuberous
eminence. The sacrum has a trapezoidal form, with callous wings and the
articulation area oriented dorsum-ventrally. The median sacral crest,
modicum, represented of 4 distanced spines.

Key words: anatomy, osteology, Carpathian bear.

The Carpathian bear is an animal with an especial cynegetic interest for


Romania. The study of the pelvis and its components constitutes an important
material that can be used in litigation situations as for the settlement of certain
morph-functional peculiarities of this species.

MATERIAL AND WORK MODE

The study has been made on bear bones provided form gunshot animals.
The bones have been prepared by boiling or by disencumbering of the adjacent
tissue.
It was pursued the shape, the lengthiness and development of bone
processes and of the articulate surfaces, in comparison with the reference
species, the particularities being illustrated by photographing.

RESULTS AND ARGUMENTS

The pelvis, as on the majority of mammals is formed of the two coxal


(ilium, ischium and pubis) and the sacral bone.
The coxal at the bear is particularized through:

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- The ilium has a wide ileac palette, oriented ventral-lateral, being provided
with a deep gluteus fosse. It has an almost triangular aspect, with a geminated
dorsal angle and at a distance of 7-12 cm. one of another. The ventral angle
presents only a tendency of germination. On the medial face of the ilium the iliac
palette is convex being provided with muscular tubercles. Auricular surface is
well delimited by a circular bony crest that stretches from the base of the iliac
palette on the body of the ilium to the neck of the ilium (fig. 1).

Fig. 1 - Dorsal view of pelvis.


A - coxal, a - ilium, b - ischium, c - pubis, 1 - wing of ilium, 2 - sacral tuber, 3 - coxal
tuber, 4 - iliac crest, 5 - incisura iliac, 6 - acetabulum, 7 - incisura ischium, 8 - ischial
tuber, 9 - ischial arch, 10 - ischial spine, 11 - obturator foramen,
B - sacrum, 1 - sacroiliac joint, 2 - spinous process, 3 - dorsal sacral foramina.
C - caudal vertebras.

The ilium crest is convex being very callous and covered in fibro-cartilage
which solidifies at the adult.
The neck of the ilium is very short, having near the circumference of the
cotillion cavity an evident tubercle for the insertion of the little tuber muscle
positioned next to the preacetabulum tubercle.
- The ischial has a massive body, the dorsal margin provides a round
ischiatic crest. The ischial tuber is very developed, in shape of a triangle, with a
tendency of splitting itself in cuspizi (ticuspida) the lateral cuspid being more
obvious and it has a slight ventral orientation.

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The caudal margin of the ischium is slightly rounded off, being covered
with a fibrocartilage partially ossified it has o dorso-ventrally, easy lateral-medial,
realizing this way a ischial arch in shape of a V with a dorsal opening.
The ischiatic table is modicum and together with the caudal portion of the
pubis forms the ischio-pubic simfiza, respective the paving of the pelvic cavity.
- the pubis has a triangular form with the cranial edge sharpened, provided
with a tuberous eminence close to the cotillion cavity that is the place for insertion
for the prepubic tendon (fig. 2).

Fig. 2 - Ventral views of pelvis


1 - auricular articular surface, 1 - scral tuber, 1 - coxal tuber, 2 - iliac crest,
3 - acetabulum tuber, 4 - transverse processes, 5 - ischial tuber, 6 - pelvic symphysis,
7 - iliopubic eminence, 8 - promontory, 9 - caudal vertebras, 10 - transverse diameter.

The cotillion cavity is deep, provided caudal-medial with a deep incision,


bordered by a strong transverse ligament, it continues with a wide rough surface
up to the center of the cavity splitting the cotillion cavity in articulate surface
and rough surface. On the anterior edge of the cavity we can see a strong
preacetabulary tuber that determines the thickening of the anterior wall of the
cavity and also the extension to the anterior of the articulate cavity (fig. 3).
The sacrum has a trapezoid form, the wings of the sacrum are thickened
having the articulate surface with the coxal oriented dorsal-ventrally, being
widespread on the entire lateral surface of it.

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Fig. 3 - Acetabulum cavity.


a - ilium, b - ischium, c - pubis
1 - ligamentary face of the acetabulary cavity, 2 - lunate articular surface, 3 - transverse
ligament, 4 - acetabulum, 5 - preacetabulary tuber, 6 - ischial spine.

The median sacrum crest is represented by 4 reduced and distanced spines,


the first having a height of 2-3 cm the rest decreasing and rounding off taking the
aspect of tubercles. Ventrally the sacrum presents 4 subsacral holes symmetrical,
wide that open up to the level of the suprasacral holes. The sagital axe is a little
curb cranial-caudal.
The diameters of the pelvis: sagital, vertical and of the caudal aperture are
almost equal.

CONCLUSIONS

1. The ilium presents a developed iliac palette, with a wide iliac fosse. The
auricular articulate surfaces are wide, set medial at the limit from the palette ant
the neck of the ilium.
2. The cotillion cavity is deep, the anterior edge is thickened and provided
anterior with a developed preacetabulary tubercle.
3. The ichiatic arcade is in a V shape, the dorsal opening and the line that
connects the ischiatic tuberoses forms an equilateral triangle.
4. The diameters of the pelvis are almost equal. The axe of the coxal is of
130.

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BIBLIOGRAFIE

1. Grasse P.P., 1972 - Traite de Zoologie, Anatomie. Systematique, Biologie. Tome


XVI, Fascic. VII, Paris.
2. Konig H.E., Liebich H.C., 1999 Anatomie der Haussngentiere. Band I,
Schattauer, Stuttgart, New York.
3. Nickel R., Schummer A., Seiferle E., 1992 Lenbuch der Anatomie der Haustiere.
Band I, Verlag Paul Parey, Berlin und Hamburg.
4. Tudor Denisa, Constantinescu Gh., 2002 - Nomina Anatomica Veterinaria. Ed.
Vergiliu, Bucureti.

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PARTICULARITI MORFO-FUNCIONALE ALE


MEMBULUI TORACIC LA VEVERI
(SCIURUS VULGARIS)
MORPHO-FUNCTIONAL PARTICULARITIES OF THE THORACIC MEMBER
ON THE SQUIRREL (SCIURUS VULGARIS)

Mihaela-Claudia SPTARU
U.S.A.M.V. Iai

Framed in the big group of the rodents, the squirrel manages to


harmonize the conformation of the skeleton in conformity with its needs for
locomotion and alimentation in the environment where it lives.
Hereby in what concerns the morpho-functional particularities of the
skeleton, on the squirrel we can observe the intermediary characteristics
between the conformations of the same structures on the rodents and cat.

Key words: scapula, humerus, squirrel.

ncadrat n grupul mare al roztoarelor, veveria reuete s-i armonizeze


conformaia scheletului conform nevoilor de locomoie i de alimentaie
dependente de mediul n care triete. Astfel, n ceea ce privete particularitile
morfo-funcionale ale scheletului, la veveri se observ caractere intermediare
ntre conformaia acelorai structuri la roztoare i pisic.

MATERIAL I METOD

Studierea particularitilor morfo-funcionale ale scheletului la veveri s-a


realizat prin disecarea i prepararea pieselor osoase raclnd esuturile adiacente
de pe acestea, urmrind lungimea i aspectul proceselor nearticulare i
ntinderea suprafeelor de articulare corespunztoare fiecrui os. Particularitile
gsite au fost fotografiate, descrise i comparate cu structurile asemntoare
ntlnite la alte specii, urmrindu-se permanent interpretarea acestora, conform
principiilor anatomice de baz, sub raportul cauz-efect.

REZULTATE I DISCUII

Membrele anterioare sunt mai scurte dect cele posterioare i sunt folosite
pentru crat i inerea hranei. Oasele centurii scapulare au particulariti
morfologice asemntoare mai mult cu cele de la feline, fiind reprezentate prin
spat i o clavicul de aproximativ 2 cm.
Spata, de form triunghiular (asemntor leporidelor) are restul
caracterelor foarte apropiate cu cele ale pisicii. Unghiul cervical este rotunjit prin

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continuarea marginii dorsale cu cea cervical, cartilajul suprascapular fiind redus


la o buz epifizar. Spina scapular este nalt, se desprinde perpendicular de pe
faa lateral a spetei pe care o mparte n cele dou fose ntr-un raport egal.
Ventral, spina scapular se termin ventral cavitii glenoide cu un acromion ce se
articuleaz cu clavicula. Paracromionul este scurt i lit.
Particularitatea spetei la veveri o constituie detaarea pe faa medial a
spetei a unei spine scapulare mediale, ce mparte suprafaa n dou fose, fosa
subscapular medial cranial i o fos subscapular medial caudal, ntr-un
raport de 1 la 3. Aceast spin ncepe i se termin la nivelul planului osos,
separnd ariile de inserie pentru musculatura subscapular, separat n dou
grupe. Procesul coracoid are aproape 0,5 cm fiind orientat caudo-ventral.
Cavitatea glenoid este ntins i pe tuberculul supraglenoidal astfel nct axul
longitudinal este de dou ori mai mare dect cel latero-medial.

Fig. 1
Fig. 2
A - Spata la veveri, faa lateral
A - Spata la veveri, faa medial
1 - unghiul cervical, 2 - spina scapular,
1 - unghiul cervical, 2 - buza epifizar,
3 - acromion, 4 - paracromion, 5 - fosa
3 - spina subscapular, 4 - proces coracoid
supraspinoas, 6- fosa infraspinoas
B Clavicula; C Humerusul;
B Clavicula; C Humerusul;
D - Radius i Ulna
D - Radius i Ulna

Humerusul este lung i rectiliniu, anul brahial este superficial, fiind


limitat de creasta deltoidien redus i de o puternic creast a epicondilului.

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Extremitatea proximal a humerusului se caracterizeaz printr-un cap


articular aproape hemisferic, mrginit de cei doi tuberculi humerali ce nu
depesc planul articular. Tuberculul mare este foarte redus, marcnd doar anul
bicipital, n schimb medial se deta un tubercul mic cruia I se pot identifica
vrful i convexitatea.
Extremitatea distal este lit transversal prin detaarea epicondililor.
ncepnd din treimea distal a humerusului se deta creasta epicondilului lateral
ca o lam osoas tioas.
Asemntor felinelor, humerusul la veveri prezint o epitrohlee masiv
cu aspect de tubercul, deasupra acesteia formndu-se gaura epitrohlear prin
detaarea unei lame osoase ce unete corpul humeral de epitrohlee.

Fig. 3 - Extremitatea distal a humerusului Fig. 4 - Extremitatea distal a humerusului


la veveri, aspect medial la veveri, aspect lateral
1 - epicondil, 2 - epitrohlee, 1 - capitulum humeri, 2 - condilul medial al
3 - creasta epicondilului, trohleei, 3 - creasta epicondilului,
4 - trohleea humeral, 4 - epitrohlee, 5 - gaur epitrohlear,
5 - fos olecranien 6 - fos radial

Articulaia cotului la veveri este n aa fel structurat nct s permit


micri ample de supinaie, asemntor felinelor, dar conserv i caractere
comune cu alte roztoare. Suprafaa articular distal a humerusului are un
caracter mixt. Lirea extremitii distale a humerusului este consecina mririi
suprafeei articulare a humerusului. Aceasta este compus dintr-un condil care
manifest caracterul de capitulum humeri numai cranial, caudal suprafaa
articular se ngusteaz, transformndu-se ntr-o creast ce delimiteaz o
veritabil trohlee. Condilul humeral se articuleaz cu cavitatea glenoid a
radiusului. Medial de condil, condilul medial al trohleei limiteaz suprafaa
articular a humerusului printr-o creast evident. Trohleea humeral devine un
an larg i superficial, perfect structurat caudal unde are o orientare oblic n sens
medial.

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Radiusul i ulna sunt lungi i uor curbate, independente i separate printr-o


arcad radio-ulnar ngust. La veveri ulna are un olecran patrulater, orientat
cranio-medial. Incizura semilunar are form de S , ntins att pe prile
laterale ale ciocului olecranului ct i pe procesul coronoid medial al ulnei.
Dintre oasele complexului carpien cel mai dezvoltat este semilunarul care
se articuleaz cu radiusul printr-un condil asemntor unui cap articular,
permind astfel micri de hiperextensie i flexie ale regiunii.
Veveria prezint cinci degete la membrul toracic, degetul I fiind redus la
un rudiment de metacarp. Metacarpienele II- V au aproximativ aceeai lungime i
aspect cu al falangelor corespunztoare proximale i medii, falanga distal este
scurt la fiecare dintre degete, fiind acoperite de gheare.

Fig. 5 - Extremitatea proximal a radiusului Fig.6 - Oasele antebraului i autopodiului


i ulnei la veveri, aspect cranial la veveri, aspect cranio-lateral
1 - olecranul, 2 - proces coronoid, 1 - radius, 2 - ulna, 3 - complexul osos
3 - incizura semilunar, 4 - cavitate carpien, I - metacarpul I, II- V - metacarpele
glenoid II-V, a - falanga I, b - falanga medie,
3 - falanga distal

CONCLUZII

1. Chiar dac este ncadrat n grupul roztoarelor, la veveri, datorit


tipului de via arboricol, oasele membrului anterior se apropie mai mult de
caracteristicile celor de la feline.
2. Caracteristic spetei de veveri este detaarea unei spine scapulare
mediale ce separ suprafeele de inserie pentru cei doi muchi subscapulari.

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3. Asemntor felinelor, humerusul la veveri are la extremitatea distal o


gaur epitrohlear i un capitulum humeri.
4. Incizura semilunar are suprafaa articular cu aspect de S, prin
ntinderea acesteia att pe prile laterale ale ciocului olecranului ct i pe
procesul coronoid lateral al ulnei.
5. Degetul I este rudimentar, metacarpienele II-V au aproximativ aceeai
lungime cu primele dou falange corespunztoare fiecrui deget.

BIBLIOGRAFIE

1. Barone R., i colab., 1973 Atlas danatomie du lapin. Ed. Masson et C-ie, Paris.
2. Coofan V., Palicica R., Valentina Hricu, Enciu V., 1999 Anatomia animalelor
domestice. vol. I, Ed. Orizonturi Universitare, Timioara.
3. Hricu Valentina, Coofan V., 2000 Anatomia animalelor de blan- Nutria i
Dihorul. Ed. Ion Ionescu de la Brad Iai.
4. Konig H.E., Liebich H.C., 1999 Anatomie der Haussngentiere. Band I,
Schattauer, Stuttgart, New York.
5. Tudor Denisa, Constantinescu Gh., 2002 - Nomina Anatomica Veterinaria. Ed.
Vergiliu, Bucureti.

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PARTICULARITI ANATOMO- FIZIOLOGICE


ALE MEMBRULUI PELVIN LA VEVERI
(SCIURUS VULGARIS)
MORPHO-FUNCTIONAL PARTICULARITIES OF THE PELVIC MEMBER
ON THE SQUIRREL (SCIURUS VULGARIS)

Mihaela-Claudia SPTARU, C. SPTARU


U.S.A.M.V. Iai

The conformation and aspect of skeleton components of the pelvic


member on the squirrel (Sciurus vulgaris) it conserves the general
characteristics met also on the rest of the representatives of the Rodentia
group. The differences demonstrated through morpho-functional
particularities, are the adaptation of the bone-ligametary and muscular
structures on the arboreal lifestyle.

Key words: squirrel, sacrum, femur, metacarp

Conformaia i aspectul componentelor scheletului membrului pelvin la


veveri conserv n mare msur caracteristicile generale ale tuturor
reprezentantelor grupului Rodenia. Deosebirile descrise ca structuri morfo-
funcionale particulare veveriei nu sunt altceva dect adaptri ale sistemului
locomotor la modul de via arboricol.

MATERIAL I METOD

Prin disecia, prepararea prin raclare i fierbere a pieselor osoase ce


compun scheletul membrelor pelvine la veveri a fost posibil evidenierea
caracterelor morfologice ale acestora. Descrierea, msurarea i fotografierea
acestora a permis efectuarea unui studiu comparativ ntre structurile osoase
provenite de la veveri (un roztor arboricol) i ale altor specii de roztoare i
carnivore cu care, datorit particularitilor de deplasare, prezint foarte multe
asemnri.

REZULTATE I DISCUII

La veveri, comparativ cu restul roztoarelor, sacrumul este format doar


din dou vertebre sudate ntre ele. Prima vertebr sacral are aceeai lime cu a
vertebrelor lombare iar datorit detarii pe toat lungimea corpului a aripilor
sacrumului acesta are aspect dreptunghiular. Aripile sunt masive, prezint pe toat
lungimea suprafaa de articulare cu coxalul care este orientat n plan vertical.
Procesele articulare craniale sunt masive i au suprafaa articular orientat

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vertical. Procesul spinos este redus la o lam scurt de aproximativ 1 mm, fiind
cel mai redus proces spinos, contrastnd cu nlimea proceselor spinoase ale
celorlaltor vertebre. Suprafaa articular a corpului vertebrei se reduce la jumtate
comparativ cu cea cranial, fiind sudate mpreun cu discul i suprafaa articular
cranial a urmtoarei vertebre sacrale.
Procesele spinoase sunt independente. Procesele mamiloarticulare se
sudeaz ntre ele delimitnd gurile suprasacrale. Procesele transverse formeaz
creasta sacrat lateral.
Cea de-a doua vertebr sacral ngusteaz pn la jumtate limea
sacrumului i are caracterele morfologice foarte asemntoare cu cele ale
vertebrelor coccigiene.
La veveri coxalul este rectiliniu i are o lungime de numai 4 - 4,5 cm.

Fig. 1 - Sacrumul la veveri, Fig. 2 - Sacrumul la veveri,


aspect latero- dorsal aspect ventral

Fig. 3 - Coxalul la veveri, aspect lateral Fig. 4 - Coxalul la veveri, aspect medial
A - Ilium, B - Ischium, C - Pubis A - Ilium, B - Ischium, C - Pubis
1 - palet iliac, 2 - creasta iliac, 1 - palet iliac, 2 - suprafa muscular,
3 tubercul muscular, 4 - cavitate 3 creasta supracotiloid, 5 - tuberozitate
acetabular, 5 - tuberozitate ischiatic, ischiatic, 6 - creasta ischiatic, 7 - simfiza
6 - creasta ischiatic, 7 - simfiza-ischio- ischio-pubien, 8 - gaura obturat,
pubien, 8 - gaura obturat 9 -suprafa articular

Poriunea preacetabular reprezint aproximativ 2 / 1 (ca la pisic, la iepure


1/ 1). Paleta iliac este dreptunghiular, reprezentnd 2/3 din lungimea total a
iliumului. Faa lateral este ornat cu o creast gluteen lung. Pe faa medial a

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paletei iliace se observ o redus suprafa articular pentru sacrum cu aspect


uniform, aezat pe gtul iliumului. Cranial de cavitatea cotiloid se deta un
tubercul sub form de creast destinat inseriei muchiului dreptul femural.
Tuberozitatea ischiatic este simpl, arcada ischiatic este larg, aproape
rectilinie, fiind cptuit de o buz epifizar ngroat. Creasta supraacetabular
este nlocuit cu o spin orientat caudal. Simfiza ischio-pubien este lung.
Femurul la veveri are aspect rectiliniu, fiind de aproximativ 5 cm
lungime. Procesele detaate la nivelul extremitii proximale ale femurului sunt
asemntoare cu cele ale leporidelor, fiind reprezentate prin capul femural i cei
trei trocamteri. Capul femural are suprafaa articular extins la de sfer, fosa
ligamentar fiind situat central. Gtul capului femural este lung, deplasndu-l
medial. Astfel, planul articulaiei coxo-femurale este plasat medial de femur i de
planul axial al acestuia, ceea ce demonstreaz c greutatea i fora este distribuit
i susinut de gtul i capul femural, fcnd posibile micri mult mai ample de
abducie i adducie necesare deplasrii. Lateral capului femural se gsete
trocanterul mare ce nu depete nlimea acestuia, avnd aspect patrulater. Sub
trocanterul mare, pn la limita distal a sfertului proximal al femurului se
desprinde sub form de creast triunghiular n unghi obtuz trocanterul al treilea.
Trocanterul al doilea apare detaat mult medial, puternic i cu aspect de triunghi
isoscel.

Fig. 5 - Femurul la veveri, aspect cranial Fig. 6 - Femur, aspect cranial


1 - capul femural; 2 - trocanter mare A - pisic; B - iepure
3 - trocanterul II; 4 - trocanterul III C - veveri
5 - trohleea femural; 6 - condil medial

Formaiunile extremitii distale a femurului la veveri se aseamn mult


cu cele ntlnite la feline, astfel trohleea femural este larg i superficial iar
condilii sunt detaai mult n sens caudal, nafara axului longitudinal al femurului,

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permind micri de extensie i flexie extrem de ample. Deasupra fiecrui condil


se gsete cte o suprafa articular pentru sesamoizii femurali.
Tibia i fibula la veveri sunt oase independente, articulate la ambele
extremiti, formnd o larg arcad tibio-fibular. La extremitatea proximal, tibia
este reprezentat prin cei doi condili care au suprafeele articulare uor excavate,
separate de spinele tibiale foarte reduse n nlime.

Fig. 7 - Tibia i fibula la veveri, aspect lateral Fig. 8 - Oasele autopodiului


membrului pelvin la veveri, aspect
dorsal
1 - tibie, 2 - fibul, 3 - oasele
tarsiene, 4 - oase metatarsiene I-V,
5 - acropodiu
a - falanga proximal, b - falanga
medie, c - falanga distal

Pe lng micrile clasice de flexie i extensie, articulaia genunchiului


poate executa micri de rotaie Creasta tibial cranial se ntinde pn la
jumtatea tibiei. Cohleea este completat lateral de maleola fibular.
La veveri membrul posterior prezint cele cinci degete cu lungime
aproximativ egal. Degetul I este format din dou falange, falangele distale sunt
scurte i acoperite de gheare puternice.

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CONCLUZII

1. La veveri sacrumul este format doar din dou vertebre.


2. Coxalul la veveri prezint caractere intermediare ntre cel de pisic i
iepure.
3. Extremitatea proximal a femurului la veveri este reprezentat printr-
un cap articular ce susine i preia greutatea i fora aplicate femurului.
4. Particularitile de conformaie ale extremitilor oaselor ce particip la
realizarea articulaiei genunchiului permit efectuarea micrilor de rotaie.

BIBLIOGRAFIE

1. Barone R., i colab., 1973 Atlas danatomie du lapin. Ed. Masson, Paris.
2. Coofan V., Palicica R., Valentina Hricu, Enciu V., 1999 Anatomia animalelor
domestice. Vol. I, Ed. Orizonturi Universitare, Timioara.
3. Hricu Valentina, Coofan V., 2000 Anatomia animalelor de blan- Nutria i
Dihorul. Ed. Ion Ionescu de la Brad Iai.
4. Konig H.E., Liebich H.C., 1999 Anatomie der Haussngetiere. Band I, Schattauer,
Stuttgart, New York, 1999.
5. Tudor Denisa, Constantinescu Gh., 2002 - Nomina Anatomica Veterinaria. Ed.
Vergiliu, Bucureti.

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PARTICULARITI ALE COLATERALELOR


I TERMINALELOR ARTERELOR ILIACE EXTERNE
LA ECVINE
PARTICULARIRIES OF SOME COLLATERALS AND TERMINAL ARTERIES
OF EXTERNAL ILIAC IN HORSE

F. STAN, A. GUDEA, A. DAMIAN, I. PAPUC


U.S.A.M.V. Cluj Napoca

The present study deals with some aspects observed during the study
of the circulatory system in horse during the normal didactical activities in
the Department of Comparative Anatomy of Faculty of Veterinary Medicine
Cluj Napoca.
Based on the dissections carried on 20 horse hind limb parts we
concluded that there is a certain amount of individual plasticity in the case
of some arteries, considered as collaterals or terminals in the pelvic and
thigh area.

Key words: anatomy, arteries, horse

INTRODUCERE

Artera iliac extern (Arteria iliaca externa) la cabaline, reprezint una


dintre terminalele aortei descendente sau caudale la aceast specie. Ea se situeaz
la marginea cranial a aperturii cavitii pelvine, naintea trunchiului venos
omonim, i parcurge un traiect uor oblic ventro-caudal, din regiunea sublombar,
pn la inelul femural, unde se termin prin artera femural. n traiectul su, emite
artera circumflex iliac profund, artera cremasteric - la mascul sau artera
uterin medie - la femel, i artera femural profund.
La rndul su, artera femural profund (Arteria femoris profunda) are
originea aproape de inelul femural i dup un traiect scurt de 4-5cm se termin
prin artera circumflex femural medial i trunchiul arterial
pudendoepigastric.
Artera circumflex femural medial (Arteria circumflexa femoris
medialis) trece pe faa medial a femurului, printre muchii pectineu i iliac,
ramificndu-se terminal n muchii caudo-laterali ai coapsei.
Trunchiul arterial pudendoepigastric (Trunchus arteriae
pudendoepigastricus) poate avea originea direct din artera iliac extern. Face o
inflexiune cranial, dorsal de inelul femural, iar la marginea cranial a inelului
inghinal profund se bifurc terminal, prin artera epigastric caudal i artera
pudend extern.

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Artera epigastric caudal (Arteria epigastrica caudalis) trece medial de


inelul inghinal profund i urmrete planeul cavitii abdominale, pe marginea
lateral a muchiului drept abdominal.
Artera pudend extern (Arteria pudenda externa) sau artera ruinoas
extern, este mai dezvoltat dect artera epigastric caudal. Ptrunde n canalul
inghinal, pe la nivelul comisurii caudale a inelului inghinal profund i se situeaz
ntre inelul i canalul vaginal pe de o parte, i inelul i canalul inghinal, pe de alt
parte. Prsete canalul inghinal la nivelul comisurii mediale a inelului inghinal
superficial, unde se termin bifurcat, prin artera dorsal cranial a penisului i
artera subcutanat abdominal, numit i arter epigastric superficial
caudal, care la masculi emite artera prepuial, iar la femele, artera mamar
cranial i artera mamar caudal.
De la nivelul inelului femural artera iliac extern se continu cu artera
femural (Arteria femoralis) care este considerat terminala sa.

MATERIAL I METOD

Lucrarea s-a efectuat n laboratorul de Anatomie comparat al facultii de


Medicin Veterinar din Cluj Napoca, pe un numr de 20 de cabaline de ambele
sexe i de vrste diferite. Acetia au constituit material didactic pentru studenii
anului II, Medicin Veterinar, n perioada 2002 2005. Ca i tehnic de lucru am
folosit: disecia, observaia, fotografia i schiele.

REZULTATE I DISCUII

n urma acestor cercetri, am observat la un individ de sex mascul, castrat,


n vrst de 12 ani, absena arterei circumflexe femurale mediale ca i terminal a
arterei femurale profunde. Topografic artera femural profund, este situat n
vecintatea inelului femural, deasupra acestuia, la civa centimetrii mai sus de
terminarea arterei iliace externe. Artera circumflex femural medial este
prezent mai jos, sub inelul femural, avnd originea n artera femural, fiind chiar
o colateral n sens caudal al acesteia. Din punct de vedere al calibrului vasului,
aceasta este mult groas comparativ cu artera femurala profund, dei ea n mod
normal (conform datelor din literatura de specialitate) ea reprezint doar una din
cele dou terminale a acesteia, cea de-a doua fiind trunchiul pudendoepigastric.
n cazul nostru, artera femural profund se confund cu trunchiul arterial
pudendoepigastric, care n toate celelalte cazuri disecate de noi, precum i coform
datelor din literatura de specialitate, ea reprezint mpreun cu artera circumflex
femural medial, terminalele acesteia. Astfel, n cazul de fa, observm c
trunchiul pudendoepigastric sau artera femural profund sunt colaterale sau mai

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bine spus rmn colaterale ale arterei iliace externe, in timp ce artera circumflex
femural medial devine colateral a arterei femurale.

Fig. 1
1 trunchiul pudendoepigastric; 2 epigastrica caudal; 3 pudenda extern;
4 circumflexa femural lateral; 5 muchiul sartorius; 6 iliaca extern;
7 inelul femural; 8 circumflexa femural medial; 9 limfocentrul inghinal profund

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Fig. 2
1 a. iliac extern; 2 trunchiul pudendoepigastric; 3 inelul femural;
4 epigastrica caudal; 5 pudenda extern

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CONCLUZII

Aceste observaii arat, c n ce privete morfologia sistemului vascular,


exist o mare variabilitate individual, att n sensul originii, ct i a traiectelor i
raporturilor anumitor vase.
n ce privete diferena de calibru ntre aceasta i artera femurala profund
sau trunchiul arterial pudendoepigastric (cazul nostru), nclinm s credem c
fiind vorba de un animal de talie mare (de traciune), este firesc, deoarece artera
circumflex femural medial reprezint unul din principalele vase trofice ale
muchilor propulsori.
Calibru mai redus a arterei femurale profunde sau trunchiului arterial
pudendoepigastric (cazul nostru) se datoreaz i faptului c este vorba de un
mascul castrat, astfel c n cazul acesta artera pudenda extern este putin mai
secundar ca importan

BIBLIOGRAFIE

1. Barone R., 1996 - Anatomie comparee des mamiferes domestique, Tome 5


Angiologie, Ed. Vigot, Paris, 687-878
2. Coofan V., Palicica R., Hricu V., Carmen G., Valeriu E., 2000 - Anatomia
animalelor domestice, vol.III., Ed.Orizonturi Universitare, Timioara, 93 - 95
3. Popovici I., Damian A., Popovici N., Chirilean I,. 2000 Angiologia animalelor
domestice, Ed.Genesis, Cluj-Napoca, 179-210
4. Pastea E., Cotofan V., Chitescu St., Miclea M., Cornila N., Nicolescu V., Radu C.,
Popovici I., Palicica R., 1985 - Anatomia comparata a animalelor domestice-
Vol. I, Ed. Didactica si pedagogica Bucuresti, 394-397
5. Damian A., 2001 - Sistemul cardiovascular, Edit. Genesis Cluj-Napoca, 111 115
6. Papuc I., Cristina Paca, Vioara Mirean, 2001 - Anatomie comparat i histologie,
Aparatul Cardiovascular, vol. I, Editura Risoprint Cluj-Napoca, pag 281 289.

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CERCETRI PRIVIND NIVELUL PLASMATIC AL UNOR


COMPONENTE BIOCHIMICE ENZIMATICE
LA OBOLANI WISTAR GREFAI IP. CU ASCITA
TUMORAL WALKER, CORELAT CU CONSUMUL
DE AP SRCIT N DEUTERIU*
RESEARCHS CONCERNING THE PLASMA LEVEL OF SOME HEPATIC ENZYMES
IN WISTAR RATS GRAFTED IP. WITH WALKER ASCITE TUMOR,
CORRELATED WITH DEUTERIUM DEPLETED WATER INTAKE*

I.B. MARCUS, S. IANCU, B. SEVASTRE


U.S.A.M.V. Cluj-Napoca

The aim of the experiment was to perform an evaluation of the plasma


level of some hepatic enzyme in Wistar rats inoculated ip. with Walker ascite
tumor, correlated with the deuterium depleted water intake. Were made up 4
groups of adults rats, each of them containing 5 animals per group. To the
groups 2 and 4 were given, ad libitum, deuterium depleted water (DDW)
daily, during 6 months. In this time, the animals were clinically evaluated and
the body weight was weekly determined. After that, the animals from the
groups 3 and 4 were inoculated ip. with Walker ascite tumor. After 4 weeks
all animals were killed, under narcose, and blood, plasma and tissues
samples were gathered. From plasma samples were measured the values of
some hepatic enzime, as ASAT, ALAT, PAL and GGT. The values recorded
for the majority of the evaluated parameters are constantly situated over the
superior limit considered normal for this species/category of animals.

Key words: Walker ascite, deuterium depleted water, enzimes.

INTRODUCERE

Utilizarea apei superuoare, cunoscut i sub numele de ap srcit n


deuteriu, n clinica oncologic dateaz din a doua jumtate secolului XX. Privite la
nceput cu suspiciune i nencredere (Redston Griffiths, 1975, Schroeter, 1992),
cercetrile ultimelor decenii realizate n acest domeniu au demostrat (Samlay i col.,
1993, 2001), n baza unor investigaii pe ct de riguroase, pe att de laborioase,
faptul c, printr-un mecanism nc incert, apa superuoar intervine n controlul
creterii tumorale spontane i experimentale. Cercetrile realizate n aceast direcie
au dus la dezvoltarea i lansarea pe piaa romneasc i strin (Samlay i col.,

*
Studiu finanat printr-un Grant de cercetare ncheiat cu CNCSIS, Tema 11, Cod CNCSIS
790. Realizarea cercetrilor a fost susinut material i de ctre RAAN, Drobeta Turnu
Severin.

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2001) a unor produse cu valoare terapeutic n oncologie, i nu numai, pe baz de


ap srcit n deuteriu, utilizate mai ales n controlul creterii tumorale la om, dar
i la animale (Bild i col., 1999). n acest moment, pe piaa romneasc exist doi
productori de ap superuoar, dintre care cel mai important este reprezentat de
RAAN (Regia Autonom pentru Activiti Nucleare), situat n Drobeta Turnu
Severin, cu sprijinul i participarea cruia a fost realizat acest studiu.

MATERIAL I METOD DE LUCRU

Experimentul a fost realizat pe un numr de 20 de obolani Wistar, aduli,


femele, care au fost distribuii n 4 loturi, a cte 5 animale per lot, dup cum
urmeaz: Lotul 1 (n=5), cu o greutate medie/animal la introducerea n experiment
de183,67,33 g/animal, a fost utilizat ca lot martor de referin. Lotul 2 (n=5), cu o
greutate medie/lot la introducerea n experiment de 214,436,3 g/animal, a fost
utilizat pentru evaluarea aciunii apei srcite n deuteriu. Lotul 3 (n=5), cu o
greutate medie/animal la introducerea n experiment de 156,211,7 g/animal, a
fost utilizat ca lot martor tumoral, inoculat cu ascita Walker. Lotul 4 (n=5), cu o
greutate medie/animal la introducerea n experiment de204,421,1 g/animal, a
fost utilizat pentru evaluarea efectului apei superuoare asupra creterii tumorale
experimentale. In acord cu protocolul experimental stabilit, animalele din loturile 2
i 4 au consumat ap srcit n deuteriu timp de 6 luni, interval dup care loturile
3 i 4 au fost inoculate intraperitoneal (ip.) cu tumora ascitic Walker. n tot acest
timp, animalele au fost urmrite clinic, nregistrndu-se lunar evoluia masei
corporale. Dup realizarea transplantului tumoral, evaluarea clinic i
determinarea greutii corporale s-a fcut sptmnal, iar dup un interval de 4
sptmni animalele au fost sacrificate prin puncia cordului, dup o prealabil
narcoz cu eter. S-au recoltat probe pentru determinri hematologice, biochimice
i gravimetrice. Din probele de plasm obinute s-a determinat concentraia
enzimelor hepatice, respectiv ALAT, ASAT, PAL i GGT. Datele obinute au fost
prelucrate statistic, prin determinarea mediei aritmetice simple (x), a deviaiei
standard (s), deviaiei procentuale (%) i a testului t de semnificaie statistic.

REZULTATE I DISCUII

n tabelul care urmeaz (tabel 1) sunt prezentate datele prelucrate statistic


ale rezultatelor obinute la sfritul experimentului, din punctul de vedere al
valorilor nregistrate n ce privete concentraia plasmatic a unor enzime cu
specificitate hepatic i tisular, respectiv transaminazele ALAT i ASAT,
fosfataza aclanin (PAL) i gamaglutamiltransferaza (GGT).

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Tabelul 1
Valori ale concentraiei plasmatice a enzimelor determinate
la loturile experimentale la sfritul experimentului

Enzimograma plasmatic
Calcul Fosfataza
ALAT ASAT
startistic alcalin GGT
GPT, ALT GOT, AST
PAL (UI/L)
(UI/L) (UI/L)
(UI/L)
Lotul 1 x 31,44 96,06 68,2 6,12
(n=5) s 15,52701 36,95623 5,514073 3,435404
x 47,26 124,76 180,5 4,14
Lotul 2 s 12,00242 12,36074 47,85269 0,971082
(n=5) D% 1 33,4744 23,00417 62,21607 -47,8261
T test 1 0,724353 0,863325 7,84E-05 0,037523
x 29,2 119,36 121,34 4,02
s 23,89236 45,62552 34,67381 2,070507
Lotul 3 D% 1 -7,67123 19,52078 43,7943 -52,2388
(n=5) D% 2 -61,8493 -4,52413 -48,7556 -2,98507
T test 1 0,424289 0,692976 0,00359 0,350319
T test 2 0,210902 0,026774 0,54794 0,171614
x 44,22 195,54 213,88 3,12
s 16,14921 97,86794 74,60166 2,513364
D% 1 28,90095 50,8745 68,11296 -96,1538
Lotul 4 D% 2 -6,87472 36,1972 15,60688 -32,6923
(n=5) D% 3 33,96653 38,95878 43,26725 -28,8462
T test 1 0,941125 0,085671 0,000176 0,559789
T test 2 0,579466 0,001464 0,410776 0,092463
T test 3 0,466687 0,168486 0,166903 0,716352

Un prim aspect care trebuie menionat este faptul c, n ce privete


corelaiile dintre consumul de ap srcit n deuteriu i particularitile
evolutive ale diferitelor forme de cancer la animale, respectiv valorile enzimelor
plasmatice investigate, n literatura de specialitate nu se gsesc date relevante n
acest sens. Din acest motiv, interpretarea rezultatelor care urmeaz a fi
prezentate i discutate va lua ca punct de referin valorile acestor componente
plasmatice la animale de laborator sntoase sau bolnave de afeciuni nrudite.
Valorile normale ale enzimelor plasmatice investigate, pentru specia de
animale i categoria de vrst pe care s-a lucrat (Cathy A. Johnson-Delaney,
1996), prezint variaii, uneori foarte largi, situate ntre 17,5 30,2 UI/L pentru
ALT, 45,7 80,8 UI/L pentru AST, i 56,8 128,0 UI/L pentru PAL.
Un prim aspect care se impune la analiza rezultatelor obinute l constituie
faptul c valorile nregistrate la lotul martor (1), utilizat ca lot martor de referin,
se situeaz n mod constant la limita superioar de oscilaie furnizat de literatura
de specialitate (Cathy A. Johnson-Delaney, 1996), i considerat a fi normal

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pentru aceast specia i categoria de animale. n raport cu aceast situaie se poate


constata faptul c, la lotul 2 (fig. 1), care a consumat zilnic ap srcit n
deuteriu, valorile ALAT, ASAT i PAL se afl mult peste limita superioar a
celor obinute la lotul de referin, i implicit a datelor furnizate de literatur,
diferena maxim fiind obinut n cazul fosfatazei alcaline (+62,2%), a crei
cretere este foarte semnificativ statistic (p<0,001). n schimb, GGT
nregistreaz o scdere a concentraiei plasmatice la 4,14 UI/L, ceea ce reprezint
o diferen de - 47,82% fa de martor, fr s aib, ns, i semnificaie statistic.

Fig. 1. Diagrama valorilor medii ale enzimelor plasmatice


determinate la loturile 1 (M) si 2 (DDW)

200
180
180,5 Lotul 1 (M)
***
160 Lotul 2 (DDW)
140
120
100
80
60
40 - 47,8%
20
0
ALAT (ui/l) ASAT (ui/l0 PAL (ui/l) GGT (ui/l)

Fig. 2. Diagrama valorilor medii ale enzimelor plasmatice


determinate la lotul tumoral (3) comparativ cu lotul de referinta (1)
ALAT (ui/l)
140
ASAT (ui/l)
120 PAL (ui/l) + 43,8%
+ 19,5% **
100 GGT (ui/l)

80

60
40
- 7,67%
20 - 52,.2%

0
Lotul 1 (M) Lotul 3 (AW)

O situaie asemntoare (fig. 2) poate fi constatat i n cazul lotului 3


(inoculat ip. cu tumora Walker, forma ascitic) care, raportat la lotul martor (1) i la
valorile furnizate de literatur, nregistreaz diferene intens pozitive pentru ASAT

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(+19,5%) i, mai ales pentru PAL (+43,6%), n timp ce valorile ALAT i GGT
scad vizibil, nregistrnd diferene procentuale de -7,67%, respectiv de -52,2%.
Determinarea comparativ a valorilor enzimelor plasmatice la loturile 3
(martor tumoral) i 4 (inoculat cu tumora Walker i care a consumat ap srcit n
deuteriu) evideniaz o situaie cel puin interesant. Astfel, dac diferenele
nregistrate n cazul lotului 3 fa de martor (1) sunt pozitive pentru ASAT i PAL,
respectiv negative pentru ALAT i GGT, pe fondul unei dezvoltri insidioase a
tumorii ascitice Walker, la lotul 4, oscilaiile nregistrate sunt i mai mari, n ciuda
faptului c lipsesc semnele clinice i necropsice care s sugereze dezvoltarea
tumoral. Practic nici unul din animalele lotului 4 nu a dezvoltat ascita Walker
inoculatat ip i, cu toate acestea, valorile enzimelor plasmatice investigate se
situeaz, constant, mult peste limita superioar a lotului martor de referin (1) i,
inclusiv a lotului martor tumoral (3), aspect care reiese cu claritate i din fig.3.

Fig. 3. Diagrama valorilor medii ale enzimelor plasmatice


determinate la loturile 3 (Walker) i 4 (DDW+Walker)

Lotul 3 (AW)
250 Lotul 4 (DDW+W)

200 +43,2%
+38,9%

150

100

50
-28,8%
+33,9%
0
ALAT (ui/l) ASAT (ui/l) PAL (ui/l) GGT (ui/l)

In sfrit, din fig. 4 rezult c, pentru toate enzimeme plasmatice determinate,


valorile i diferenele procentuale nregistrate comparativ cu lotul martor sunt, n
cazul loturilor 2, 3 i 4 constant pozitive pentru ASAT i PAL, constant negative
pentru GGT, i cu o evoluie inconstant pentru ALAT. Diferenele cele mai
pregnante afecteaz, i n cazul lotului 4, ASAT (+50,8% fa de lotul martor, i
+38,9% fa de lotul 3), fosfataza alcalin (+ 68,1% fa de lotul 1, i +43,26% fa
de lotul 3), respectiv GGT (-96% fa de lotul martor i 28,8% fa de lotul 3).
Raportat la aceste rezultate, trebuie menionat c un nivel plasmatic crescut
al unor enzime cum sunt alanin-amino-transferaza (ALAT), aspartat-amino-
transferaza (ASAT), fosfataza alcalina (PAL) i gama-glutamil-transferaza (GGT)
la animale poate fi rezultatul unor modificri/leziuni reversibile sau ireversibile
ale membranelor hepato-celulare, asociate cu aspecte de necroz hepatic,
colestaz, hipoxie sau inflamarea ficatului determinat de agenii infecioi sau

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toxine bacteriene. Ct privete specificitatea modificrii valorii acestor


componente biochimice plasmatice, ALAT este considerat o enzim specific
ficatului, dei este prezent n concentraie redus i n alte esuturi, iar ASAT
este, de asemenea, prezent n concentraii mari n hepatocite, astfel nct trebuie
urmrit atent n cazul n care aceasta crete fr s antreneze o cretere
corespunztoare a ALAT. Referitor la PAL i GGT, acestea sunt enzime asociate
membranei celulare, iar creterea concentraiei lor plasmatice este asociat cu
leziuni ale zonei pericanaliculare structurilor biliare (Ghergariu i col., 2000).

Fig. 4. Diagrama valorilor medii ale enzimelor plasmatice


determinate la loturile experimentale luate n studiu.
220
210
200 Lotul 1 (M)
190
180 Lotul 2 (DDW)
170
160 Lotul 3 (AW)
150 Lotul 4 (DDW+W)
140
130
120
110
100
90
80
70
60
50
40
30
20
10
0
ALAT (ui/l) ASAT (ui/l) PAL (ui/l) GGT (ui/l)

CONCLUZII

Concluziile care se desprind din analiza i discutarea rezultatelor prezentate


mai sus, coroborate cu informaiile furnizate de literatura de specialitate privitor la
valorile normale i situaiile clinice care antreneaz modificri ale concentraiei
enzimelor plasmatice investigate, au n vedere urmtoarele aspecte principale:
1) Concentraiile plasmatice ale enzimelor studiate (ASAT, ALAT, PAL i
GGT) n cadrul acestui experiment la loturile martor se ncadreaz n limitele de
oscilaie considerate normale pentru specia i categoria de animale pe care s-a
lucrat, cu specificarea c ele se situeaz uor peste sau sub limita superioar.
2) Este, practic, imposibil de precizat, n acest stadiu al experimentului,
dac consumul ndelungat de apa srcit n deuteriu (DDW) este responsabil de
creterea semnificativ a concentraiei enzimelor investigate, avnd n vedere
diferenele foarte mari nregistrate comparativ cu lotul martor de referin.
3) n acelai timp, este dificil de explicat cauza pentru care la lotul martor
tumoral (3), inoculat ip. cu tumora ascitic Walker, valorile nregistrate se

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

ncadreaz n limitele de oscilaie considerate normale normale n cazul ALAT,


PAL i GGT, i sunt doar foarte uor crescute fa de normal n cazul ASAT.
4) Considerm, de asemenea, mai mult dect interesant, asocierea celor mai
crescute (n cazul ALAT, ASAT, PAL), respectiv mai sczute (n cazul GGT)
valori ale enzimelor investigate la lotul 4 (inoculat ip. cu tumora ascitic Walker i
care a consumat ap srcit n deuteriu), mai ales n condiiile n care la nici unul
din animalele acestui lot nu au fost evideniate aspecte ale dezvoltrii tumorale.

BIBLIOGRAFIE

1. Bild W., Stefanescu I., Haulica I., Lupusoru C., Titescu G., Iliescu R., Nastasa V.,
1999 - Research concerning the radioprotective and immunostimulating effects
of deuterium-depleted water, Rom J Physiol. Jul-Dec;36 (3-4):205-218.
2. Ghergariu S., Marina Spanu, Pop Al., Kadar L., 2000 - Manual de Laborator Clinic
Veterinar, Ed. All Educational, Bucuresti, 34-67.
3. Joseph J. Katz, 1965 - Chemical and Biological Studies with Deuterium, Thirty-Ninth
Annual Priestley Lectures, Pennsylvania State University, 26-29.
4. Kritchevsky D., 1960 - Deuterium Isotope Effects in Chemistry and Biology Annals
of the New-York Academy of Science vol. 84, New-York: Academy of Science.
5. Lamprect J., 1990 - Mitosis Arrested By Deuterium Oxide light microscopic,
immunofluorescence and ultrastructural characterization European Journal of Cell
Biology 51: (2) 303-312, Wissen-schaftliche Verlag GMBH.
6. Redston Griffiths T., 1975 - The Possible Roles of Deuterium in the Initiation and
Propagation of Aging and Other Biological Mechanisms and Processes,
Proceedings of the Second International Conference on Stable Isotopes, Oak
Brook Illinois, October 20-23.
7. Samlay G., 1993 - Naturally occurring deuterium is essential for the normal growth
rate of cell, FEBS Lett.,vol 317, Nr.1,2,1-4,februarie, SAMLAY si colab., The
biological effect of deuterium-depleted water, a possible new tool in cancer
terapy.,Z. Onkol. / J.of Oncol, 30:4,1998.
8. Samlay G, 2001 - The biological effect of deuterium-depleted water, a possible new
tool in cancer therapy. Anticancer Research, 21:1617-1621,
9. Schroeter D., 1992 - Deuterium Oxide Arrests the Cell-Cycle of PTK2 Cells During
Interphase European Journal of Cell Biology 58:(2) 365-370 Stuttgart, Germany:
Wissenschaftliche Verlag MBH.

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INVESTIGAII BIOCHIMICE PRIVIND VALORILE


TRANSAMINAZELOR SERICE I ALE
COLESTEROLEMIEI CORELATE CU SENESCENA
LA CINI CLINIC SNTOI
BIOCHEMICAL INVESTIGATIONS REGARDING THE VALUES
OF SERIC HEPATIC ENZYMES AND CHOLESTEROL CORRELATED
WITH AGING IN CLINICALLY HEALTHY DOGS

S. IANCU, I.B. MARCUS


U.S.A.M.V. Cluj-Napoca

The aim of the study was to reveal some clinical and para-clinical
modifications that appear with the installation and evolution of aging in
clinically healthy dogs. The study was conducted on 24 individuals, male
and female dogs, with age ranges between 8 and 15 years, fed and raised as
pets, in apartment conditions. The subjects were divided in two groups:
group 1 (n - 12) with an age range between 8 and 10 years, and group 2 (n
- 12) with an age range between 10 and 15 years. Following, every group
was divided on genders males and females, so that 4 groups resulted, as
mentioned below. Blood samples were collected from all 4 groups, and with
a bicromatic fotometer, the hepatic enzymes (ALAT, ASAT), GGT, PAL and
cholesterol level were determined for the evaluation of modifications that
appear with aging. The obtained results revealed that the value of the
investigated parameters increases together with the aging process. A part of
the obtained values is situated in normal ranges or slightly above the
average of animals from groups 1 and 2 (age between 8 and 10 years), but
for groups 3 and 4 (age between 10 and 15 years), the obtained values are
constantly situated above the normal average of this species.

Key words: dogs, aging, clinically healthy, blood biochemistry.

INTRODUCERE

mbtrnirea sau senescena reprezint un fenomen biologic natural,


ireversibil, care afecteaz lumea vie n ansamblul ei, constnd n procese de uzur
la nivel celular, tisular i sistemic care premerg moartea celulei, esuturilor i, n
final a organismului (Paolisso i col., 2000; Morley, 2004). Manifestarea cea mai
pregnant a mbtrnirii o constituie declinul progresiv al funciei celulelor,
esuturilor i organelor, nsoit de accentuarea producerii de radicali liberi i
fenomene de liz celular, disfuncii mitocondriale, dezechilibre endocrine i
instabilitate genomic (Mac Lean i col., 2004; Irina M. Conboy i col., 2005).
Capacitatea regenerativ a esuturilor este mult afectat odat cu naintarea

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animalelor n vrst, iar acest lucru se manifest la nivel sanguin prin tulburri
care afecteaz funcia hematopoietic a mduvei osoase i, n acelai timp, prin
modificri ale valorilor unor parametri celulari, biochimici i enzimatici serici
(Mac Lean i col., 2004). Astfel, creterea sau scderea valorilor unor enzime
serice poate constitui un indiciu valoros n aprecierea sensului i a severitii
tulburrilor paraclinice care nsoesc instalarea i evoluia procesului de senescen
la animale (Morley, 2004). n acest sens, creterea nivelului seric al unor enzime
cum sunt fosfataza alcalina (PAL), gamaglutamiltransferaza (GGT), alanin-amino-
transferaza (ALAT) i aspartat-amino-transferaza (ASAT) la cine poate fi
rezultatul: 1) schimbrilor reversibile sau ireversibile ale membranelor hepato-
celulare, asociate cu necroza hepatic, colestaz, hipoxie, hipoperfuzia sau
inflamarea ficatului determinat de agenii infecioi, toxine sau de mbtrnire; 2)
din depozite excesive de lipide (ficat gras), de cupru sau glicogen n hepatocit. Ct
privete specificitatea acestor componente biochimice serice, ALAT este
considerat o enzim specific ficatului la cine i pisic dei ea este prezent n
concentraie redus i n alte esuturi, iar ASAT este, de asemenea, prezent n
concentraii mari, astfel nct trebuie urmrit atent n cazul n care aceasta crete
fr o cretere corespunztoare a ALAT. PAL i GGT sunt enzime asociate
membranei celulare. Inducia enzimatic apare, n cazul lor, nainte de a se realiza
creteri importante n snge. Aceste enzime sunt asociate cu leziuni ale zonei
pericanaliculare structurilor biliare i reticului endoplasmatic. Hipercolesterolemia
apare la pacieni suferinzi de: ocluzia cilor biliare, diabet, pancreatite,
hipotiroidism, hiperadrenocorticism, sindrom nefrotic, hiperlipidemie postprandial,
hipelipidemia idiopatic a Schnauzerului (Hostkins, 2004).

MATERIAL I METOD DE LUCRU

Studiul a fost realizat pe animale cu vrste cuprinse ntre 8 i 15 ani, care au


fost mprite n 4 loturi, dup cum urmeaz: Lotul 1 (n=7) i Lotul 2 (n=5) alctuite
din femele i masculi de rase diferite, cu vrste cuprinse ntre 8-10 ani; Lotul 3
(n=7) i Lotul 3 (n=5) alctuite din femele i masculi de rase diferite, cu vrste
cuprinse ntre 11-15 ani. Toate animalele din cele 4 loturi au fost supuse unui
minuios examen clinic n prealabil, n urma cruia nu a fost depistat nici un semn
clinic care s sugereze prezena sau evoluia unei mbolnviri, cu excepia faptului
c o parte din indivizi examinai au fost supraponderali, iar din anamnez s-a
desprins faptul c marea majoritate aveau o alimentaie dezechilibrat.
Determinrile biochimice efectuate din probele de ser recoltate de la
animalele luate n studiu au fost realizate cu ajutorul unui fotometru bicromatic de
uz general, STAT FAX 1904 PLUS, compact, controlat de un microprocesor cu 6
filtre i bloc de incubare la 37C. Aparatul poate fi utilizat pentru a msura
absorbane sau concentraii bazate pe reacii cinetice sau cu punct final. Orice test
care necesit citire la lungimile de und identice sau apropiate cu cele ale filtrelor
din dotare, pot fi efectuate cu ajutorul aparatului. Aparatul are inclus un bloc de
incubare cu 12 poziii. Ca principiu de lucru, fasciculul de lumin de la o lamp cu

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halogen trece mai nti printr-o lentil, apoi este direcionat printr-o apertur, iar n
final traverseaz orizontal prin prob. Valorile obinute n urma determinrilor
efectuate la cele 4 loturi de animale luate n studiu au fost prelucrate statistic,
determinndu-se media aritmetic, deviaia standard i diferena procentual.

REZULTATE I DISCUII

n tabelul 1 sunt prezentate limitele de oscilaie furnizate de literatura


(Ghergariu i col., 2000; Marcus, 2004) pentru toi componeni biochimici i
enzimatici luai n studiu, calculndu-se pentru fiecare parametru media
aritmetic, n vederea raportrii i interpretrii statistice a rezultatelor obinute la
loturile de animale investigate.
Tabelul 1
Valori normale ale parametrilor investigai
(Ghergariu i col., 2000; Marcus, 2004)

Parametrul COLES-
ALAT ASAT GGT PAL
biochimic TEROL
(U/l) (U/l) (U/l) (U/l)
Valori (mg/dl)
Valori de referin 20
5 102 10 55 10 15 173
(limite de oscilaie) 156
X 53,5 32,5 12,5 88 173

Analiza rezultatelor obinute la cele 4 loturi de animale luate n studiu a


evideniat urmtoarele aspecte clinice i paraclinice: la examenul clinic toate
animalele prezentau semne mai mult (cele din categoria 10-15 ani) sau mai puin
(categoria 8-10 ani) accentuate ale instalrii fenomenelor de mbtrnire. n ce
privete determinrile biochimice, pentru majoritatea parametrilor investigai,
limitele de oscilaie se ncadreaz n limite normale pentru aceast specie de
animale (Ghergariu i col., 2000; Marcus, 2004), cu cteva variaii mai
semnificative fa de valorile de referin, care privesc, la lotul 1 (tabel 2 i fig. 1),
scderea ALAT (-40,95%), creterea valorii GGT (-53,51%) i PAL (+43,05%)
inclusiv creterea colesterolemiei (+40,95%). Situaia se menine asemntoare i
n ce privete lotul II (n=5), n componena cruia intr masculi cu vrsta cuprins
ntre 8 i 10 ani, i la care nivelurile serice obinute pentru componentele
investigate sunt i mai apropiate de cele considerate normale pentru aceast specie
de animale, exceptnd i n acest caz uoara scdere a ALAT (-17,22%) i
creterea cu 27,09 procente a fosfatazei alcaline (PAL).

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Tabelul 2
Valori ale parametrilor investigai la lotul I () (8 -10 ani)

Parametrul
COLES-
biochimic Calcul ALAT ASAT GGT PAL
TEROL
statistic (U/l) (U/l) (U/l) (U/l)
(mg/dl)
Cazul
X 37.96 39.07 8.14 154.5 292.97
LOTUL 1
(n=7) +S 14.30 13.04 3.04 33.16 149.74
D% -40.95 16.82 -53.51 43.05 40.95

Fig. 1. Diagrama valorilor medii obinute la loturile 1 i 2


(8-10 ani) comparativ cu date le de refe rin din lite ratur
160 Valori de referinta 154,51
140 Valori lotul 1(n=7 )
Valori lorul 2 (n=5 ) 120,7
120
100

80

60

40 37,96
39,07 52,56
20 45,64 8,14 12,64
0
ALAT ASAT GGT PAL

Tabelul 3
Valori ale parametrilor investigai la lotul II () (8-10 ani)

Parametrul
COLES-
biochimic Calcul ALAT ASAT GGT PAL
TEROL
statistic (U/l) (U/l) (U/l) (U/l)
(mg/dl)
Cazul
X 45.64 52.56 12.64 120.7 165.46
LOTUL II
(n=5) +S 32.96 8.85 2.98 50.09 42.89
D% -17.22 38.17 1.11 27.09 -4.56

Semnificaia diferenelor statistice obinute n cazul valorii componentelor


serice investigate la loturile 3 () i 4 (), n componena crora intr animale cu
vrsta cuprins ntre 11 i 15 ani (tabel 4 i 5, fig. 2), se amplific n cazul tuturor
parametrilor.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Fig. 2. Diagrama valorilor medii obinute la loturile 3 i 4


(11-15 ani) comparativ cu datele de referin

180 176,36
Valori de referinta
160 160,8
Valori lotul 1(n=7 ?)
140 Valori lorul 2 (n=5 ?)
120
100
80 82,44
64,12 60,26
60
40 44,51
20 10,41 17,52

0
ALAT ASAT GGT PAL

Tabelul 4
Valori ale parametrilor investigai la lotul III () (11 -15 ani)

Parametrul
COLES-
biochimic Calcul ALAT ASAT GGT PAL
TEROL
statistic (U/l) (U/l) (U/l) (U/l)
(mg/dl)
Cazul
X 44.51 82.44 10.41 160.9 250.53
LOTUL III
(n=7) +S 21.93 71.46 3.49 70.98 93.60
D% -20.19 60.58 -20.03 45.31 30.95

Astfel, comparativ cu valorile de referin (tabel 1) furnizate de literatura de


specialitate (Ghergariu i col., 2000; Marcus, 2004), n cazul cinilor cu vrsta
cuprins ntre 11 i 15 ani diferenele procentuale (%) nregistreaz variaii pozitive
pentru ASAT (+60,58%), PAL (+45,31%) i Colesterol (+30,95%), respectiv
negative pentru ALAT (-20,19%) i GGT (-20,03) n cazul lotului 3.
Tabelul 5
Valori ale parametrilor investigai la lotul IV () (11 -15 ani)

Parametrul
COLES-
biochimic Calcul ALAT ASAT GGT PAL
TEROL
statistic (U/l) (U/l) (U/l) (U/l)
(mg/dl)
Cazul
X 64.12 60.26 17.52 176.36 205.2
LOTUL IV
(n=5) +S 39.15 25.61 13.08 10.58 74.51
D% 16.56 46.07 28.65 50.10 15.69

n cazul lotului IV, constituit din masculi cu vrsta cuprins ntre 11 i 15


ani, pentru toate componentele investigate, diferenele procentuale nregistreaz
variaii pozitive, mai mult sau mai puin accentuate, exprimate prin creterea

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valorii serice a ALAT (+16,56%), ASAT (+46, 07%), GGT (+28,65%), PAL
(+50,1%) i, inclusiv Colesterol (+15, 69%).
Fig. 3. Diagrama valorilor medii ale colesterolemiei la loturile de
animale investigate, comparativ cu valorile de referin
Valori de referinta
Lotul I
Lotul II
300 Lotul III
292,97 Lotul IV
250 250,53
200
205,2
150 173 165,46
100

50

0
Colesterolul (mg/dl)

Dac pentru majoritatea parametrilor enzimatici determinai diferena %


fa de valorile de referin de ncadreaz n limitele de oscilaie normale, uneori
cu uoare depiri ale limitei superioare, n cazul colesterolemiei, valorile medii
obinute i, implicit diferenele % nregistrate se situeaz cu mult peste media
normal de 173 mg/dl. Ceea ce rezult din analiza informaiilor prezentate i
discutate mai sus este c, dac din punct de vedere clinic instalarea senescenei se
exprim prin modificri vizibile i relativ uor de cuantificat la un examen
obiectiv al animalelor, n ce privete tabloul paraclinic corelat acestui fenomen,
lucrurile nu mai sunt att de limpezi. Considerat din aceast perspectiv,
mbtrnirea este un fenomen lent, progresiv, care, n primele faze (considernd
vrsta de 8-10 ani la carnasiere) nu se exprim prin modificri semnificative la
nivelul componentelor biochimice i enzimatice serice, acestea instalndu-se abia
peste limita de 10 ani, i doar la intensiti care nu depesc dect rareori limita
superioar considerat normal pentru specia de animale investigat.
Foarte probabil c semnificaia biologic redus a variaiilor nregistrate
pentru parametri determinai se datoreaz tocmai unor limite de oscilaie extrem
de largi, care merg de la 5 la 102 UI n cazul ALAT, sau de la 20 la 156 UI n
cazul fosfatazei alcaline (PAL), pentru ca n cazul altor enzime serice (ex: GGT),
aceste limite de oscilaie normale s se situeze constant ntre 10 i 15 UI.
Interesant de remarcat este i faptul c, n cazul unora din componentele serice
determinate, valorile medii situate mult peste limita superioar considerat
normal se coreleaz cu o dispersie foarte larg a valorilor individuale, exprimate,
de exemplu, prin valori ale deviaiei standard de pn la 149,74 n cazul
colesterolului la lotul I, 50,09 n cazul PAL la lotul II, 71,46 i 70,98 n cazul
ASAT i PAL la lotul III, respectiv 74,51 n cazul colesterolemiei la lotul IV.

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CONCLUZII

Analiza rezultatelor prezentate i discutate mai sus, coroborate cu datele din


literatura de specialitate, permit cteva concluzii cu privire la urmtoarele aspecte:
1) Senescena este un fenomen biologic care se instaleaz progresiv, antrennd,
ntr-o prim etap modificri exprimate printr-un tablou clinic specific;
2) Din punct de vedere paraclinic, la animalele vrstnice, clinic sntoase,
tabloul biochimic sanguin nu nregistreaz modificri notabile, cel puin n faza
incipient a mbtrnirii, considerat n cazul prezentului studiu, la vrsta de 8-10 ani.
3) n faze mai avansate ale mbtrnirii (11-15 ani), specificitatea tabloului
clinic se coreleaz i cu alterri ale tabloului biochimic i enzimatic sanguin,
exprimate prin scderea sau creterea concentraiei unor componente la acest nivel.
4) Variaiile cele mai semnificative afecteaz, n acest caz, valorile
colesterolemiei, care nregistreaz constant nivele situate mult peste media
considerat normal la cine.
5) Nu se poate constata o specificitate determinat de sex n ceea ce
privete sensul i severitatea tulburrilor nregistrate, respectiv tipul componentei
biochimice afectate.

BIBLIOGRAFIE

1. Ghergariu S, Marina Spanu, Pop Al., Kadar L., 2000 - Manual de Laborator Clinic
Veterinar, Ed. All Educational, Bucuresti, 34-67.
2. Irina M. Conboy, Michael J. Conboy, Amy J. Wagers, Eric R. Girma, Irving L.
Weissman, Thomas A. Rando, 2005 - Rejuvenation of aged progenitor cells by
exposure to a young systemic environment; Nature, Vol 433, 17.
3. Ittai Ben-Porath, Robert A Weinberg, 2004 - When cells get stressed: an
integrative view of cellular senescence. Journal of Clinical Investigation. Ann
Arbor: Vol. 113, Iss.1; 8-13.
4. Jeffrey A. Klein and Susan L. Ackerman, 2003 - Oxidative stress, cell cycle, and
neurodegeneration; J. Clin. Invest. 111:785-793.
5. John E. Morley, 2004 - The Top 10 Hot Topics in Aging; The Journal of Gerontology;
Vol. 59, Iss. 1; pg. 24-33.
6. Johnny D. Hostkins, 2004 - Geriatric and gerontology of the dog and cat, Saunders,
56-87.
7. Jolly C. A., 2004 - Dietary Restriction and Immune Function; J. Nutr. 134: 1853-56
8. Marcus I.B, 2004 - Biologia i Patologia Animalelor de Laborator, Edit. Risoprint,
Cluj-Napoca, 156 -178.
9. Mc Lean AL and David G. Le Couteur, 2004 - Aging Biology and Geriatric Clinical
Pharmacology; Pharmacol. Rev. Vol. 56 Nr. 2:163-184.
10. Olovnikov A.M., 2003 - The redusome hypothesis of aging and the control of
biological time during individual development; Biochemistry (Mosc); 68(1):2-33.
11. Paolisso G, M. Barbieri, M. Bonafe, C. Franceschi, 2000 - Metabolic age mode-ling:
the lesson from centenarians; European Journal of Clinical Investigation, 30, 888-
894
12. Pop Al., I.B.Marcus, 2004 - Fiziopatologie, Edit. Risoprint, Cluj-Napoca, 36-68.

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EVALUAREA EFECTULUI CHEMOPREVENTIV AL UNOR


PLANTE APARINND FAMILIEI LAMIACEAE
PARTEA 1: APRECIEREA EFECTULUI
CHEMOPREVENTIV-HEPATOPROTECTOR AL UNOR
FITOPREPARATE PRIN PRISMA ALANIL
AMINOTRANSFERAZEI SI ASPARTAT
AMINOTRANSFERAZEI
THE CHEMOPREVENTIVE EFFECT EVALUATION OF SOME PLANTS FROM
LAMIACEAE FAMILY
PART 1: THE CHEMOPREVENTIVE-HEPATOPROTECTOR EFFECT ESTIMATION OF
SOME PHYTOPREPARATIONS BY POINT OF VIEW OF ALANIL
AMINOTRANSFERASE AND ASPARTAT AMINOTRANSFERASE

Cornelia PRISCARU
U.S.A.M.V- Iai
Anca-Irina BURLACU
U.M.F.Gr. T. Popa Iai

The experiment reproduced in this work aims at finding of some


plants with active elements which are able to exercise a chemopreventive
action against the hepatotoxic aggression of the various xenobiotics. The
experiment directed its attention on some plants of Lamiaceae family,
starting from specialty literature data which relieve the chemopreventive
effect of the rosemary (Rosmarinus officinalis) at the exposure with the
carcinogen mycotoxins with difuranic structure. The experiment included 5
lots of the white rats, Wistar line: the first lot represented the reference lot,
while the second lot was used for the experimental reproduction of the
intoxication with sterigmatocystin, furo-furanic mycotoxin. The three last
lots had the designation to detectable the existence of the eventual
chemopreventive effect at the rosemary, garden sage (Salvia officinalis) and
lavender (Lavandula angustifolia) on the background of the intoxication
with sterigmatocystin. The estimate of the chemopreventive-antitoxic action
made by some biochemical parameters which offer the informations about
the hepatocit integrity: alanil aminotransferase and aspartat
aminotransferase.

Key words: sterigmatocystin, chemopreventive, Rosmarinus officinalis,


Salvia officinalis, Lavandula angustifolia

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INTRODUCTION

Vegetal systematic studies which are correlated with the existence of the
therapeutic virtues conduct at the conclusion that plants of the same family or
order manifest farmacognostic similitudes. The rosemary (Rosmarinus officinalis,
Lamiaceae family), prevalent appreciated plant for its alimentary value, is
imposed at the end of the last century by the antitoxic effect (1). The merit to
attract the attention on this aspect belongs to some mycotoxicologists who relieve
the rosemary property to stimulate the synthesis of an enzyme implicated on the
secondary metabolism action of the aflatoxins, the mycotoxins which are
considered by IARC uman carcinogens of the rank I. The active principle of the
rosemary leaves structure, capable to induce the synthesis of glutation-S-
transpherase is a ditiol-tione, named by who that put it in the evidence oltipraz
(2). In this way the xenobiotics of the difuranic mycotoxin type are facile excreted
in the shape of glutationo-conjugates, preventing the formations of the adducts
between the reactive form of the mycotoxin and the elite molecules of the
molecule (AND, ARN, enzymes, a.s.o.), which constitutes the biochemical lesion.
Starting from these arguments, this work proposes to verify the possibility of the
chemopreventive/antitoxic effect existence at the plants on systematic criteria
(Lavandula angustifolia and Salvia officinalis). For the chemopreventive action
test of the vegetal products of the respective plants it was selected as the toxicity
standard sterigmatocystine, the ubiquitous mycotoxin in the geographical area
with the temperate-continental climate in which our country is also situated. The
chemical and biogenetic relation of the aflatoxin, the sterigmatocystine exercises
its toxic effects at the hepatic level, where it is subdued to the metabolism action,
so as it results from the fig. 1 (3).

Fig. 1 - The primary transformation of the sterigmatocystine

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MATERIAL AND METHOD

For to detect as well as possible the virtual chemopreventive intervention of


the vegetal products, it was imagined an experimental model which to watch the
parameters with the biggest relevance for the hepatocit integrity (table 1). The
experiment included 5 lots of the white rats Wistar 7 months old and 189.26g
medium weight. The first lot, formed of 5 rats , represented the reference lot, was
fed with the standard fodder and maintained in the standard conditions of habitat.
The second lot, also formed of 5 animals, was the lot on which it reproduced the
intoxication with sterigmatocystine. In this way the animals of this lot were daily
treated with 8 ppm sterigmatocystine. The third lot (5 animals), concomitant with
the dose of 8 ppm sterigmatocystine, was also given fitopreparat 5% of
Rosmarini folium, ad libitum. The animals of the fourth lot (5 animals),
concomitant with the dose pro die of mycotoxin difuranic, was also given extract
5% of Salviae herba. The last lot of the experiment, formed of 5 white rats, served
for the estimate of the virtual chemopreventive effect of the lavender flowers.
Consequently, the animals of this lot were treated with 8 ppm sterigmatocystine
and phytopreparat 5% de Lavandulae flores, in the drunken water ad libitum. The
experiment carried on the period of 6 weeks, when the animals were treated
according to the scheme of the experimental model (table 1). At the end of these
6 weeks the animals were sacrificed, and the gathered blood was biochemical
examined. The transaminaze serice activity (ALT and AST) was performed by
Reitman-Frenkel method, with the fixed time of the incubation (4,5).

Table 1
Experimental model

Sterigmatocystine Rosmarini Salviae Lavandulae Biochemical


LOTS
[ppm] folium 5% herba 5% flores 5% parameters
Lot 1 - - - -
Lot 2 8 - - -
Lot 3 8 ad libitum - - AST, ALT
Lot 4 8 - ad libitum -
Lot 5 8 - - ad libitum
Legend:
AST = aspartat aminotransferase
ALT = alanil aminotransferase

RESULTS AND DISCUSSIONS

The evolution of the aspartat aminotransferase activity, so it results of the


table 2, makes evident a significant development in the serum of the intoxicated
lot with mycotoxin furo-furanic (131.38 UI), in comparison with the witness lot
(54.67 UI). For the treated lots with the phytopreparations on the basis of the

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rosemary and the garden sage, it registers the lightly grown values, but though
closed of the reference lot values (57,94 UI, respectively 65.69 UI), what it
suggests the favorable intervention of the active principles from the respective
plants. Not the same evolution registers the enzyme activity for the treated lot
with the lavender extract, its value being about 133.92 UI, what it makes evident
grave affectations of the hepatocit integrity. Referring to the alanil
aminotransferase activity, which is in accordance with the table 2, it may come to
the conclusion of the existence of a citolize hepatic for the exclusively treated lot
with sterigmatocystine (45.40 UI in comparison with only 17.50 UI of the witness
lot), but also for the protected lots with the phytopreparations on the basis of the
garden sage (50.95 UI) and the lavender (46.42 UI). Only the value ALT of the lot
which profited of the exogenous contribution of the rosemary phytopreparation
indicates a hepatic protection, its activity being about 19.2 UI, the very closed
value of the witness lot (17.50 UI).

Table 2
Transaminase evolution

AST ALT
LOTS [ UI ] [ UI ]
Minimum Medium Maximum Minimum Medium Maximum
Lot 1 48.15 54.67 68.30 20.38 17.50 23.50
Lot 2 100.50 131.38 142.50 40.50 45.40 49.51
Lot 3 49.50 57.94 66.30 15.40 19.20 25.95
Lot 4 45.95 65.69 88.60 33.40 50.95 88.15
Lot 5 120.00 133.92 148.90 38.90 46.42 55.41

CONCLUSIONS

The aspartat aminotransferase evolution makes evident the existence of a


chemopreventive-antitoxic potential of the active principles from the rosemary
and the garden sage;
The aspartat aminotransferase oscilation does not suggest the appearance of
a chemopreventive-antitoxic effect exercised by the lavender;
The alanil aminotransferase activity confirms the antitoxic action of the
active principles from Rosmarini folium;
The alanil aminotransferase evolution refutes the existence of the
chemopreventive potential at the phytopreparations both at the lavender and the
garden sage.

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BIBLIOGRAPHY

1. Istudor Viorica, 2001 Farmacognozie, Fitochimie, Fitoterapie, Vol. II, Editura


Medical, Bucureti, 122;
2. Kensler T.W., 1997 Chemoprevention by inducers of carcinogen detoxification
enzymes, Environ Health Perspect., 965-970;
3. Priscaru Cornelia, 1998 Cercetri privind incidena micotoxinelor n produse de
origine vegetal i animal din zona Podiului Moldovei i mecanismele de
toxicocinetic i toxicodinamie ale acestora, Tez de doctorat, UMF Gr.
T.Popa, 188;
4. Filip Maria, Pduraru Ioana, aramet I., 1993 Biochimie-Lucrri practice, Editura
Gr. T. Popa, Iai, 61-68, 105-107;
5. Mihele Denisa, Pavlovici M., 1996 Biochimie clinic, Editura Medical, Bucureti,
99-103.

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EVALUAREA EFECTULUI CHEMOPREVENTIV AL UNOR


PLANTE APARINND FAMILIEI LAMIACEAE
PARTEA 2: APRECIEREA EFECTULUI
CHEMOPREVENTIV-HEPATOPROTECTOR AL UNOR
FITOPREPARATE PRIN PRISMA ACTIVITII -
GLUTAMIL TRANSPEPTIDAZEI SI COLINESTERAZEI
THE CHEMOPREVENTIVE EFFECT EVALUATION OF SOME PLANTS
FROM LAMIACEAE FAMILY
PART 2: THE CHEMOPREVENTIVE-HEPATOPROTECTOR EFFECT ESTIMATION OF
SOME PHYTOPREPARATIONS BY POINT OF VIEW OF -GLUTAMIL
TRANSPEPTIDAZE AND CHOLINESTERASE

Cornelia PRISCARU, Liliana ROTARU


U.S.A.M.V. Iai

The present work represents a stage of a more detailed experiment


which has mission to discover the virtual chemopreventive action against
the hepatotoxic aggression of the various xenobiotics. There were aimed at
the plants of Lamiaceae family due to some bibliographical details which
relieve the chemopreventive effect of the rosemary (Rosmarinus officinalis)
at the exposure with the carcinogen mycotoxins with difuranic structure
and due to their relation with this plant. The experiment includes 5 lots of
the white rats, Wistar line. The first lot represented the reference lot (the
control lot), while the second lot served for the experimental reproduction
of the intoxication with sterigmatocystin, hepatotoxic mycotoxin, chemical
related with the aflatoxins. The three last lots served for to verify the
existence of the eventual chemopreventive effect at the rosemary
(Rosmarinus officinalis), garden sage (Salvia officinalis) and lavender
(Lavandula angustifolia) on the background of the intoxication with
sterigmatocystin. The estimate of the chemopreventive-antitoxic action
made by some biochemical parameters with the relevance for the integrity
and hepatic function: -glutamil transferaze and seric cholinesterase.

Key words: hepatoprotector effect, Rosmarinus officinalis, Salvia officinalis,


Lavandula angustifolia

INTRODUCTION

The rosemary (Rosmarinus officinalis, Lamiaceae family), the known plant


for its quality to spice of the aliments, but also its therapeutic virtutes (1),
surprises the scientific world of the mycotoxicology domain in the end of the last
century by the antitoxic effect (2). It was proved that the daily administration of

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some capable substances to induce the synthesis of some enzymes implicated in


the second stage of the xenobiotic metabolism action at the liver level, prevents
the formations of the mycotoxin-ARN, mycotoxin-AND, mycotoxin-protein
adducts with high specialized function, the adducts which constitutes the
biochemical carcinogen lesion (3). The rosemary contains in its leaves a ditiol-
tion with the chemopreventive action at the treatment with the daily doses of the
aflatoxin B1 (2). These reasons constituted the arguments on which is based the
present work which tried to verify the possibility of the
chemopreventive/antitoxic effect existence at the plants which are related with the
rosemary, namely the lavender (Lavandula angustifolia) and the garden sage
(Salvia officinalis). For the chemopreventive action estimate of the vegetal
products of the respective plants it was selected as the toxicity standard
sterigmatocystine, the ubiquitous mycotoxin in the geographical area with the
temperate-continental climate in which our country is also situated.

MATERIAL AND METHOD

The experimental model on which is appeared this work is presented in the


table 1. This included 5 lots of the white rats Wistar 7 months old and 189.26 g
medium weight.
Table 1
Experimental model

Sterigmatocistine Rosmarini Salviae Lavandulae Biochemical


LOTS
[ppm] folium 5% herba 5% flores 5% parameters
Lot 1 - - - -
Lot 2 8 - - -
Lot 3 8 ad libitum - - GGT, ChE
Lot 4 8 - ad libitum -
Lot 5 8 - - ad libitum
Legend:
GGT = -glutamil transpeptidaze
ChE = cholinesterase

The first lot, formed of 5 rats, represented the reference lot, was fed with
the standard fodder and maintained in the standard conditions of habitat. The
second lot, also formed of 5 animals, was the lot on which it reproduced the
intoxication with sterigmatocystine. In this way the animals of this lot were daily
treated with 8 ppm sterigmatocystine. The third lot (5 animals), concomitant with
the dose of 8 ppm sterigmatocystine, was also given phytopreparat 5% of
Rosmarini folium, ad libitum. The rats of the fourth lot (5 animals), concomitant
with the dose pro die of sterigmatocystine, was also given extract 5% of Salviae

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herba. The fifth lot, formed of 5 white rats, served for the estimate of the possible
chemopreventive effect of the lavender flowers. The animals of this last lot were
treated with 8 ppm sterigmatocystine and phytopreparat 5% of Lavandulae flores,
in the drunken water ad libitum. The experiment carried on the period of 6 weeks,
when the animals were treated in accordance with the scheme of the experimental
model (table 1), after that they were sacrificed, and the gathered blood was
biochemical examined. The -glutamil transpeptidaze activity was quantized by
the colorimetric method with -glutamil-p-nitroanilide. For the estimate of the
seric cholinesterase activity utilized the standard method with the acid 5.5-
ditiobis-2-nitrobenzoic (Ellman reagent) (4, 5, 6).

RESULTS AND DISCUSSIONS

The biochemical investigations, consist of the determination of the seric


cholinesterase activity and -glutamil transpeptidaze activity, are presented in the
table 2 and fig. 1. Watching the cholinesterase activity, the synthesized enzyme in
the liver, the possessor of a slow turnover, we find an evident diminution of this
for the exclusively treated lot with the mycotoxin, its value being about 190.08
UI, by comparison with 235.97 UI, the witness lot value. This dramatic
diminution of this biochemical parameter activity is the confirmation of the
proteosynthesis capacity affecting of the attacked liver by the mycotoxin
presence. The quantification of this parameter in the serum of the protected
animals by the phytopreparation administration of Rosmarini folium, makes
evident a value which is significantly closed of the registered value at the witness
lot (227.75 UI).
Table 2
Cholinesterase and -glutamil transpeptidaze evolution

ChE GGT
LOTS [ UI ] [ UI ]
Minimum Medium Maximum Minimum Medium Maximum
Lot 1 228.10 235.97 258.88 3.35 5.30 12.6
Lot 2 158.37 190.08 226.69 16.00 19.00 23.60
Lot 3 199.55 227.75 295.65 4.25 5.50 7.12
Lot 4 180.19 190.94 213.00 4.25 5.29 6.95
Lot 5 185.25 191.95 200.65 11.55 19.86 29.35

Although this lot was subjected to the impact with the sterigmatocystine,
the proteinic synthesis capacity of this lot was not affected, what it suggests the
rosemary intervention by the chemopreventive effect. A complete distinct
situation was registered for the two last lots, on which was verified the antitoxic
effect of the garden sage and the lavender. It may observed that in the treated lot
with the garden sage the seric cholinesterase value is 190.94 UI, and at the lot

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what was subdued to the exogenous contribution of the lavender


phytopreparation, the enzyme activity oscillates about the value 191.95 UI, what
it conducts to the conclusion of the chemopreventive effect absence of the active
principles from this two plants which are related to the rosemary. Referring to the
-glutamil transpeptidaze, the biochemical parameter used as the enzymatic
induction marker, the tumor marker and also the hepatotoxic lesion indicator, we
may find, as it results from the table 2 and fig. 1, an increase about 4 times greater
at the lot subdued to the sterigmatocystine intoxication (19 UI), by comparison
with the witness lot (5.30 UI). The similar values to the witness lot registers GGT
from the serum of the animals which profited of the exogenous contribution of the
phytopreparations obtained from Rosmarini folium and Salviae herba on the
background of the intoxication with sterigmatocystine, the values which confirm
their hepatoprotecting effect. Not the same situation may be described in the case
of the lot treated, concomitant with the mycotoxin, with the extract of Lavandulae
flores, the GGT activity from the animals serum of this lot being of 19.86 UI, the
characteristic value of a toxic lesion from the intoxication with the furanic
mycotoxin.

250
Lot 1
200 Lot 2
150 Lot 3
[UI]

Lot 4
100 Lot 5

50

0
ChE GGT

Fig. 1 - Cholinesterase and -glutamil transpeptidaze evolution


(the graphic of the above table)

CONCLUSIONS

The seric cholinesterase activity for the exclusively treated lot with the
sterigmatocystine suggests a grave affectation of the proteosynthetic function of
these animals liver;

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The seric cholinesterase level for the animals which were treated
concomitant with the mycotoxin and the phytopreparations from the three plants
makes evident the exercise of the chemopreventive effect only for Rosmarinus
officinalis;
The -glutamil transpeptidaze evolution from the serum of the exclusevily
treated animals with the sterigmatocystine registers a significant increased value;
The GGT oscillation suggests the positive intervention of the
phytopreparations on the basis of Rosmarini folium and Salviae herba;
The GGT value from the serum of the animals treated with the Lavandulae
flores invalidates the existence of a chemopreventive effect of the active
principles from this plant.

BIBLIOGRAPHY

1. Grigorescu, E., Lazr, M.I., Stnescu Ursula, Ciulei, I., 2001 Index fitoterapeutic,
Editura Cantes, Iai, 421;
2. Kensler, T.W., 1997 Chemoprevention by inducers of carcinogen detoxification
enzymes, Environ Health Perspect., 965-970;
3. Mace, K., Offord, E. A., Harris, C.C., 1998 Development of in vitro models for
cellular and molecular studies in toxicology and chemoprevention, Arch. Toxicol.
Suppl., 227-236;
4. Filip Maria, Ioana Pduraru, aramet I., 1993 Biochimie-Lucrri practice, Editura
Gr. T. Popa, Iai, 61-68, 105-107;
5. Mihele Denisa, Pavlovici M., 1996 Biochimie clinic, Editura Medical, Bucureti,
99-103;
6. erban, M., Cmpeanu, G., Ionescu, E., 1993 Metode de laborator n biochimia
animal, EDP, Bucureti, 198.

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ANALYSIS OF THE NATURAL AND INDUCED


ECHINOCYTOSIS FOR TWO SPECIES OF MAMMALS
Servilia OANCEA1, Dorina CREANGA2,
Alina RAPA3, Geta PAVEL1
1
U.S.A.M.V. Iai, Romnia
2
Univ. Al.I.Cuza, Iai, Romnia
3
West Vancouver Secondary School, BC, Canada

The normal erythrocyte shape is that of a biconcave discocyte. The


relationship between cell surface area and volume determines the extent to
which a cell can deform. Abnormalities of red cell shape can be easy
classified according to the degree of echinocytic or stomatocytic
transformation but are difficult to quantify. Traditional mechanical and
mathematical methods have proved to be insufficient for describing the
echinocytosis process. Over the last several years, fractal analysis has been
applied with great success to many biological systems. Many physiological
systems have been found to be both spatial and temporal fractals. In this
work we studied the morphological changes of erythrocytes determining the
fractal dimension for the contour of the erythrocyte. We suggested that, if
we develop an easy method to measure the images fractal dimension,
fractal dimension may be a good measurement of echinocytosis.

Key words: red blood cell deformability, HarFA soft, echinocyte contour

INTRODUCTION

The mechanical properties of erythrocytes are greatly influenced by the


membrane, especially by its cytoskeletal structure and by geometrical factors. The
RBC membrane is viscoelastic; thus the viscous component limits the rate of
deformation in addition to the elastic moduli, which mainly determine the extent
of deformation. Both the viscous and the elastic components are determinants of
deformability and primarily depend on the protein cytoskeleton.
Geometrical factors affecting red cell deformability are the size, shape and
surface area/volume ratio of the cell. The normal erythrocyte shape is that of a
biconcave discocyte. The relationship between cell surface area and volume
determines the extent to which a cell can deform. Thus an excess of surface area
over that needed to enclose the cell volume is needed for deformation. The normal
red cell has an excess area of about 50%. Any loss of surface or increase in
volume will tend to restrict the ability of the cell to pass through a narrow
aperture. It is well established that this shape represents equilibrium between two
opposing shape changes.
Abnormalities of red cell shape can be easy classified according to the
degree of echinocytic or stomatocytic transformation but are difficult to quantify.

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The data indicate that erythrocyte shape changes have an important


influence on blood rheology [2]. Discocytes had higher oxygen transport
efficiency than either echinocytes or stomatocytes. Echinocytosis increased
viscosity especially at low shear rates and may be explained by increased cell-cell
interaction, i.e. the tangling of the spicules of echinocytes. At high shear rates the
spiculated surface of echinocytes is at least partly smoothed. During the
echinocytic transformation the content of inorganic ions change.
There are different methods to measure the individual cell deformability.
Noji and coworkers [3] estimated the average of whole cell deformability using
electron paramagnetic resonance (EPR). The authors showed that the decrease in
the cell deformability was often found in haematological disorders involving
erythrocytes abnormalities (sickle cell anaemia, pregnancy) and also in non-
haematological diseases (for example diabetics, hypertension etc). In [4] was
investigated the effect of different lipid A concentration on RBC deformability.
Morphology of RBC was studied on video recordings and to characterise
echinocytes and spherocytes, the criteria established by Becker and Bessis [5].
In [6] is presented influence of nitrovasodilators on rheology of blood.
Treatment of whole blood with sodium-nitroprusside (SNP) at 10-3 M induced a
market echinocytosis and increased blood viscosity at a high shear rate and a low
shear rate whereas NO(nitric oxide) do not affect RBC shape and blood viscosity.
Donath and co-workers [7] studied the spicula distribution during echinocytosis
using a histogram of the Voronoi froth.
Traditional mechanical and mathematical methods have proved to be
insufficient for describing the echinocytosis process.
The fractal theory has been well utilised in many domains of physics and
biology [8-10]. It is possible, therefore, by using fractals to study and quantify in
a much more concise way and much less approximately those structures which
were described as granulous, tentacular, ramified, etc.
Over the last several years, fractal analysis has been applied with great
success to many biological systems. Many physiological systems have been found
to be both spatial and temporal fractals. Fractal theory permeates the study of the
cardiovascular system both in physiological and pathological conditions. Fractal
images permit to analyse the regional flow of the pulmonary parenchyma, and to
quantify pulmonary damage from thromboembolism. Kang et al. [11] found by
analysing the aggregation images that RBC aggregation shows fractal
characteristics. Their research presents the time dependence of the information
dimension for RBCs for human blood samples.
In some recent works we also applied the fractal analysis to research the
fractal structure of biological objects. In [12] the fractal analysis upon various
forestry arbour leaves was carried out in order to find the fractal dimension in
each case and We found that average values of fractal dimension are increased
when the complexity degree of leaf shape is increasing (from poplar to ash tree

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and to false acacia etc.). In [13] the fractal anlysis, was applied to the study of
erythrocytes and Jurkat cell aggregates (lymphoblastoid T-cell line). The method
revealed the fractal character of blood cell with modified shape because of
relatively high concentrations of urea and LPC (lysophosphatidylcholine). Fractal
dimension was found proportional to the urea and LPC concentration or to the
time of cell aggregation.
In this work we studied the morphological changes of erythrocytes
determining the fractal dimension for the contour of the erythrocytes.
HarFA soft was utilised to visualise the fractal dimension. In HarFA a
modification of traditional Box Counting is used. By this modification one
obtains 3 fractal dimensions, which characterise the properties of black plane DB,
the properties of white background DW and the black-white border of black
object DBW, which is the most interesting information.

MATERIALS AND METHOD

Samples from peripheral sheep blood and cow blood were performed using
May-Grwald Giemsa colorature. By means of the microscope we obtained many
photos; two of these photos from every species of mammals are presented in
Fig.1 and Fig.2

Fig.1 - Blood sample from sheep blood Fig.2 - Echinocytes from cow blood

RESULTS AND DISCUSSION

From our photos we selected 10 blood cells for the two species of mammals
in order to determine the fractal dimension of their contour. Our results for fractal
dimension of the echinocyte contour from sheep and cow blood are presented in
the table below.

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Nr. Fractal dimension (sheep) Fractal dimension (cow)


1 1.0367 1.0556
2 1.0705 1.1123
3 1.0210 1.0768
4 1.0143 1.0631
5 1.0402 1.1092
6 0.9999 1.0800
7 1.0304 1.1092
8 1.0207 1.1094
9 1.0358 1.0341
10 1.0256 1.1088
Average 1.02951 1.08585
Standard deviation 0.01873 0.028
Relative error 1,81% 2.57%

From this table we can see that the fractal dimension from cows cells is
higher than from sheep cells (the average value for cow cells is 1.08585 and for
sheep cells it is 1, 01873). Generally speaking in the sheep sample there is no
echinocytosis because the fractal dimension is near 1. For cow blood
echinocytosis is induced by a disease.

CONCLUSION

We can suppose that if we develop an easy method to measure the images


fractal dimension, fractal dimension may be a good measurement of
echinocytosis. The fractal analysis can be a method to find the stage of
echinocytosis, measuring the fractal dimension from the contour of the cells. A
disease bring about the echinocytosis in cow blood. Measuring the degree of
echinocytosis we can suggest then the disease that produces this process.
Weiss and co-workers [15] evaluated echinocytosis in both healthy and sick
dogs and they concluded that two diseases, glomerulonephritis and
lymphosarcoma may be associated with this phenomenon. The shape of the
erythrocytes can also be altered by a great variety of chemical agents, such as
many detergents due to their amphiphilic nature [16]. The crenated erythrocytes
from animals exposed to detergent appeared either with border irregularities or
undulations, without distinct spicules, or with numerous short spikes. In [17] Geor
and co-workers showed that the most frequent diagnosis for horses, which present
echinocytosis, was colitis and they concluded that hyponatremia was associated
with increased incidence of echinocytosis. They suggested that systemic
electrolyte depletion might be involved in the induction of echinocyte formation.
To evaluate the hypothesis that echinocytes are dehydrated erythrocytes, in [18]
the authors decreased blood sodium and potassium concentrations in 4 horses by
administering furosemide.

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Due the fact that the average value of the fractal dimension is not very high
compared with 1, which is for normally blood, we suggest that the cow is
experiencing the first fase of the disease.

REFERENCES

1. Shiga T., Maeda N., Kon K., 1990 - Erythrocyte Rheology, Oncology/Hematology,
Vol.10. Issue 1, 9-48.
2. Reinhart W.H., Singh-Marchetti M., Straub P.W., 1992 - The influence of
errythrocyte shape on suspension viscosities, Eur. J. Of Clinical Investigation,
22, 38-44.
3. Noji S., Taniguchi S., Kon H., 1991 - An EPR Study on Erythrocyte deformability,
Progr.Biophys. molec. Biol., Vol. 55, pp. 85-105.
4. Pschl J.M.B., Linderkamp O., 1992 - Effect of lipid A on the deformability,
membrane rigidity anf geometry of human adult red blood cells, Eur. J. Of
Clinical Investigation, 22, 625-629.
5. Bessis, M., Weed, R.,J. - Red cell shape, Springer Verlag,Berlin, New
6. Walter R., Mark M., Gaudenz R., Harris L.G., Reinhart W.H., 1999 - Influence of
nitrovasodilators and endotelin-1 on rheology of human blood in vitro, British J.
Of Pharmacology, 128, 744-750.
7. Donath E., Pomorski T., Pastushenko F., 1993 - On the lateral distribution of
spicula on echinocytes, Biorheology, 30, 463-470
8. Nonnenmacher T.F., Losa G.A., Weibel E.R, 1993 - Ed., Fractals in Biology and
Medicine, Boston, USA.
9. Nonnenmacher T.F., Baumann G, Barth A, Losa GA, 1994 - Digital image analysis
of self-similar cell profiles, Int J Biomed Comput , 37(2), 131-138
10. Guarini G, Onofri E., 1993 - New horizons in medicine. Fractals, Recenti Prog Med.,
84(6), 438-42
11. Grasman J., Brascamp J.W., Van Leeuwen J.L., Van Putten B., 2003 - The
multifractal structure of arterial trees, J Theor Biol, 220 (1), 75-82
12. Kang M.Z., Zeng Y.J., Liu J.G., 2000 - Fractal research on red blood cell
aggregation, Clin. Hemorheol. Microcirc. 22, 229-236
13. Creanga I., Oancea S., Tudorie M., Creanga D., Bara I., 2002 - Oak leaves
fractality, Int. J.of Chaos Theory and Application, 7 (1-2), 85-90
14. Creanga D., Tudorie M., Oancea S., Rapa A., 2003 - Semi-quantitative analyse of
cells and cell aggregates, Sci. Ann. USAMV Iasi, XLVI, v. 2, Proceedings of the
Annual Symposium on Mathematics applied in Biology & Biophysics, Univ. of
Agricultural Science and Veterinary Medicine, Iasi, 293-298
15. Weiss D.J., Kristensen A., Papenfuss N., McClay C.B., 1990 - Quantitative
valuation of echinocytes in the dog, Vet Clin Pathol., 19(4), 114-118
16. Zeni C., Bovolenta M.R., Stagni A., 2002 - Occurrence of echinocytosis in
circulating RBC of black bullhead, Ictalurus melas (Rafinesque), following
exposure to an anionic detergent at sublethal concentrations, Aquat Toxicol,
57(4), 217-24
17. Geor R.J., Lund E.M., Weiss D.J., 1993 - Echinocytosis in horses: 54 cases (1990),
J Am Vet Med Assoc. 202(6), 976-80.
18. Weiss D.J, Geor R.J., Smith C.M., McClay C.B., 1992 - Furosemide-induced
electrolyte depletion associated with echinocytosis in horses, Am J Vet Res., 53
(10), 1769-72

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SPECTROPHOTOMETRIC ANALYSIS
OF THE BLOOD PLASMA
Servilia OANCEA, Iuliana MOTRESCU,
U.S.A.M.V. Iai
lioancea@univagro-iasi.ro
imotrescu@yahoo.com
Alina RAPA,
West Vancouver Secondary School, Canada
alinarapa@yahoo.com
A. AIRINEI
Institute of Macromolecular Chemistry P. Poni,
Iasi, Romania
airineia@icmpp.ro

In this paper an analysis of the blood plasma spectrum is performed


in order to establish the relationship between protein concentration and
their absorbance in the UV region. We found a direct dependence of these
quantities correlating the spectra and the refractive index measured for the
same protein concentrations. This suggests that the specific properties and
protein concentration of plasma influence the blood behavior.

Key words: absorption spectra, protein band, refractive index

INTRODUCTION

Blood plasma, a yellow fluid contains proteins, nutrients, metabolic end


products, hormones, and inorganic electrolytes. It is a very complex and not fully
understood mixture of proteins that performs and enables many housekeeping and
other specialized bodily functions.
In blood plasma, by far the most prevalent protein is albumin,
approximately 32 to 35 grams per liter, which helps to maintain osmotic balance
of the blood. Many studies reported quantitative results on blood plasma
properties for human samples [1, 2].
The spectrophotometric analysis of the blood plasma was used to
investigate small molecules in human plasma and determine protein concentration
using FTIR [3]. Human plasma retinol-binding protein and transthyretin was also
studied [4]. Fluorescence characteristics of blood plasma were studied in the
visible spectral region, at two different excitation wavelengths, 405 and 420 nm,
to discriminate patients with different stages of oral malignancy from healthy
subjects [5]. The authors showed that the fluorescence spectra of blood plasma of
oral malignant. subjects exhibit characteristic spectral differences with respect to

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normal subjects. Analysis of blood plasma protein is also used to study the
Alzheimer's disease [6].
There is a little data on plasma properties and protein content for different
animals. Specific properties of plasma of different mammalian species influence
the blood behavior.
In this work we studied the characteristic spectrum of the blood plasma,
which is determined by the protein absorbance band in the UV domain in order to
determine the concentration of protein.

MATERIALS AND METHOD

To analyze the blood plasma spectrum, blood samples were collected in


test tubes containing EDTA as an anticoagulant. About 20 ml of blood from cow
and horse are sampled for this test.
The sampled blood is centrifuged to separate blood plasma (4000 g during
5 minutes). Then we prepared different solutions from blood plasma in distilled
water.
Measurements of plasma refractive index were performed with an Abbe
refractometer.
Spectrophotometric analysis in UV and Visible domains of the blood
plasma were performed with a spectrophotometer Specord M-42.

RESULTS AND DISCUSION

Fig. 1 and Fig. 2 present the characteristic spectrum of the blood plasma in
the VIS and UV domains.

Fig. 1. Plasma spectrum in VIS domain (horse blood)

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Fig. 2. Plasma spectrum in UV domain (horse blood)

The protein absorbance band in the UV domain (about 280 nm) determines
the characteristic spectrum of the blood plasma.
The value of the absorption band strongly depends on the blood plasma
concentration. This dependence is presented in fig. 3.

0,6
y = 0,0031x + 0,0599
0,5 R2 = 0,9772

0,4
absorbance

0,3

0,2

0,1

0
0 50 100 150
concentration of plasma (*10-4)

Fig. 3. Plasma absorption at 280nm versus concentration

We observed that the protein absorption at 280nm exhibit a good linear


dependence on the concentration as well as the refractive index as shown in fig. 4.

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1,355
y = 0,0174x + 1,332

refractive index
1,35
R2 = 0,9895
1,345
1,34
1,335
1,33
0 0,5 1 1,5
concentration of plasm a

Fig. 4. Plasma refractive index versus concentration

CONCLUSION

The present study suggests that the spectrophotometric analysis of the


blood plasma is a useful tool for determination plasma protein content and also is
an easier method than the other ones. Due the fact that the protein absorbance
band at 280nm determines the characteristic spectrum of the blood plasma and the
value of the absorption band strongly depends on the blood plasma concentration,
this method permits the determination of a small content of protein from plasma
blood. This means we can use a very small quantity of plasma, which is diluted in
distilled water.

REFERENCES

1. Narayahan, S., Galloway, L., Nonoyama, A., Leparc, G.F., Garcia-Rubio, L-H.,
Potter, R.L., 2002 - UV-visible spectrophotometric approach to blood typing II:
phenotyping of subtype A2 and weak D and whole blood analysis, Transfusion,
42(5), 619-626
2. Zahao, B., Tham, S.Y., Lu, J., Lee, L., Moochhala, S., 2004 - Simultaneous
determination of vitamins C, E and -carotene in human plasma by high-
performance liquid chromatography with photodiode-array detection, J
Pharmaceut Sci, 7(2), 200-204
3. Petibois, C., Cazorla, G., Cassaigne, A., Deleris, G., 2001 - Plasma Protein
Contents Determined by Fourier-Transform Infrared Spectrometry, Clinical
Chemistry, 47, 730-738
4. Raghu, P., Ravinder, P., Sivakumar, B., 2003 -A new method for purification of
human plasma retinol-binding protein and transthyretin, Biotechnology and
Applied Biochemistry, 38, 19-24
5. Mahduri, S., Vengadesan, N., Aruna, P., Koteeswaran, D., Venkatesan, P., Ganesan,
S., 2003 - Native Fluorescence Spectroscopy of Blood Plasma in the Characterization
of Oral Malignancy, Photochemistry and Photobiology, 78 (2), 197-204
6. Ueno, I., Sakai, T., Yamaoka, M., Yoshida, R., Tsugita, A., 2000 - Analysis of blood
plasma proteins in patients with Alzheimer's disease by two-dimensional
electrophoresis, sequence homology and immunodetection, Electrophoresis,
21(9), 1832-1845.

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THE HEMATOLOGICAL PROFILE OF LIMOUSINE


HEIFERS DURING THE ADAPTATION-
ACCLIMATIZATION PERIOD
PROFILUL HEMATOLOGIC LA JUNINCILE DE RAS LIMOUSINE
N PERIOADA DE ADAPTARE-ACLIMATIZARE

C. FALC,, Gh. CIORBA, Cristina PETRUSE

inndu-se cont de faptul c sngele prin constituenii si celulari i


biochimici, constituie oglinda sntii animalului (reacionnd rapid i de
multe ori specific la agresiunea determinat de factorii interni sau externi),
am optat n principal pentru realizarea unor investigaii de ordin
hematologic, att din punct de vedere cantitativ, ct i calitativ la junincile
de ras Limousine clinic sntoase aflate n perioada de adaptare-
aclimatizare.
Investigaiile paraclinice se refer la junincile de ras Limousine
importate n ara noastr n anul 2002, de ctre o unitate de producie din
zona de vest a rii i la produii acestora testai n anul 2005. Rezultatele
obinute au fost comparate cu cele provenite de la junincile mame din anul
2002, care au fost recent aduse n ar.
Parametri hematologici (leucocite i trombocite) au avut o valoare
mai sczut cu aproximativ 16 % la junincile din anul 2005 fa de
junincile din anul 2002, diferenele nefiind ns asigurate statistic. Pentru
eritrocite valorile medii au fost aproape identice (p > 0,01).
Hemoglobina, hematocritul, VEM-ul, HEM-ul i CHEM-ul au
prezentat valori mai mari la junincile din anul 2002 comparativ cu junincile
din anul 2005, diferenele variind ntre 0,95 % i 14,36 %. La aceste valori
diferenele ntre cele dou perioade sunt nesemnificative.

Key words: Limousine heifers, blood, hematological parameters

Pathology`s current orientation to know the metabolic health status aims at


an as quick as possible discovery of the critical metabolic area between normal
and abnormal, the prevention of the status of clinical disease, the establishment of
the limits concerning the physiological oscillations of the various metabolites
within the internal environment. Such variations represent undoubtedly a useful
step to achieve data useful to make possible in the future to prevent some
dysmetabolic affections (2, 3). Under intensive systems, nutrition and metabolism
diseases interfere into breeding, and, through their frequently clinical evolution,
dysmetabolic affections represent an important cause for economic losses (1).
The major objective of this paperwork is to establish reference data
regarding the hematological profile of Limousine heifers, recently imported in our
country, and also its evolution during the adaptation-acclimatization period.

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MATERIAL AND METHOD

The hematological investigations were carried out on 11 Limousine heifers


from a farm located in the Western part of the country.
Blood samples were taken from the jugular vein (about 3-4 ml, on EDTA
anticoagulant). Immediately after the sample taking, we sent them to laboratory
for an analysis with the hematological analyzer MS 9 VET. In order to assess
properly the erythrocytary system in the case of the studied individuals, we
determined the direct erythrocytary constants (erythrocyte number, hematocrit,
hemoglobin) and the indirect erythrocytary constants (VEM, HEM, CHEM).
The completion of the hematological examination consisted of
investigations upon the leukocytary series and of the determination of
thrombocyte number.
The survey upon the white cellular series was carried out through
quantitative determinations the determination of the total leukocyte number, and
qualitative the percentage rhyming of the different categories of leukocyte from
the leukocytary formula.
The obtained data was processed through biostatistic methods, using the
software Excel for calculations. In order to test the difference significance we
used the test Mann-Withney.

RESULTS AND DISCUSSIONS

Analyzing the information within table 1, we observe that in heifers (2002)


the number of erythrocytes was 8.74 0.42 mil/mm3, and in heifers (2005) it was
almost like that from the first investigation 8.62 0.46 mil/mm3. We do not
observe any significant difference between these values.
Hemoglobin concentration in heifers from 2002 was 12.39 0.62 g/dl, and
in those from 2005, 11.47 0.93 g/dl. In the case of this parameter, no significant
difference is revealed between the two investigations, too.
The hematocrit in heifers from 2002 had a value of 47.99 2.28%,
respectively 46.14 1.30 % in heifers from 2005. Any significant difference is
not observed between the two animal groups.
Regarding VEM, there is not any significant difference between the values
obtained in heifers from 2002 (54.96 1.12 3) and those from 2005 (54.44
2.18 3). HEM had the value of 14.17 0.40 pg in heifers from 2002, and 13.19
0.53 pg in heifers from 2005, there being not any significant difference.
CHEM was 25.79 0.34 g/dl in heifers from 2002 and 22.55 1.87 g/dl in
those from 2005. There is not any significant difference concerning this constant
between the two paraclinical investigations. We mention that VEM and CHEM
values are according to the reference values from the literature of speciality.

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Table 1
The evolution of hematological parameters in heifers (n = 1)
and the significance of the differences

Differences Significances
Specification X S x Cv% (Mann-
abs % Withney)
Heifers 2002 8,74 0,42 12,60 + 0,12 1,39 1,0000
Erythrocytes
Heifers 2005 8,62 0,46 14,07
Heifers 2002 12,39 0,62 13,17 + 0,92 8,02 0,5229
Hemoglobin
Heifers 2005 11,47 0,93 21,46
Heifers 2002 47,99 2,28 12,52 + 1,83 3,96 0,8480
Hematocrit
Heifers 2005 46,14 1,30 7,46
Heifers 2002 54,96 1,12 5,38 + 0,52 0,95 0,5656
VEM
Heifers 2005 54,44 2,18 10,59
Heifers 2002 14,17 0,40 7,42 + 0,98 7,42 0,4432
HEM
Heifers 2005 13,19 0,53 10,64
Heifers 2002 25,79 0,34 3,47 + 3,24 14,36 0,3379
CHEM
Heifers 2005 22,55 1,87 21,94

In Table 2, we present the values of the hematological parameters in heifers


and the difference significance. We might note that in the case of the heifers
(2002) the number of leukocytes was 5.92 0.64 thousand/mm3, compared to
heifers (2005) 5.08 0.45 thousand/mm3, without any significant difference
between the two periods.
Table 2
The values of hematological parameters in heifers (n = 1)
and the significance of the differences

Differences Significances
Specification X S x Cv%
abs %
(Mann-
Withney)
Heifers 2002 5,92 0,64 28,48
Leukocytes +0,84 16,53 0,3379
Heifers 2005 5,08 0,45 23,45
Heifers 2002 61,77 1,01 4,32
Lymphocytes -0,54 0,86 0,6547
Heifers 2005 62,31 3,49 14,78
Heifers 2002 6,90 0,77 29,30
Monocytes -0,67 8,85 0,4062
Heifers 2005 7,57 1,20 42,01
Heifers 2002 31,24 1,00 8,45
Granulocytes +1,13 3,75 0,8480
Heifers 2005 30,11 2,44 21,36
Heifers 2002 268,43 30,52 30,02
Thrombocytes +38,43 16,70 0,3066
Heifers 2005 230,00 24,37 27,98

Regarding lymphocyte number, the obtained values are 61.77 1.01 % in


the case of the heifers (2002), respectively 62.31 3.47 % in heifers (2005).
Monocyte number in heifers (2002) was 6.90 0.77, respectively 7.57
1.20 in heifers (2005). There are no any significant differences from a statistic

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point of view. Granulocytes had almost the same values in heifers (2002) 31.24
1.00 %, respectively 30.11 2.44 % in heifers (2005).
Thrombocyte number was 268.43 80.52 thousand/mm3 in heifers (2002),
respectively 230.00 24.37 thousand/mm3 in heifers (2005).

CONCLUSIONS

The values of the hematological parameters (leukocytes and thrombocytes)


are lower with about 16% in the heifers from 2005 compared to those from 2002,
the differences being not statistically supported.
For lymphocytes, monocytes and granulocytes, the differences are
insignificant between heifers from 2005 compared to those from 2002, the lowest
difference appears in the case of lymphocytes 0.86%.
Regarding erythrocytes, the average values are almost identical (p > 0.01).
Hemoglobin, hematocrit, VEM, HEM and CHEM have higher values in the
heifers from 2002 compared to those from 2005, the differences varying between
0.95% and 14.36%. But the differences between the two periods are insignificant
at these values.

BIBLIOGRAPHY

1. Falc, C., 2004 Semiologie medical veterinar, vol II, Ed. Cosmopolitan Art,
Timioara.
2. Ghergariu, S., 1995 - Patologia nutriional i metabolic a animalelor, Ed. Medical
Veterinar, Bucureti.
3. Prvu, Gh., 1992 Supravegherea nutriional metabolic a animalelor de ferm. Ed.
Ceres, Bucureti.
4. Reibel, Ch., 2003 Charollais et Limousine en haut des preferens, Reussin Bovins
Viande, 99.
5. Stanciu, G., Acatinci, S., Cziszter, L., 1994 Tehnologia creterii bovinelor, Ed.
Euroart, Timioara.

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THE EFFECTS OF CALCIUM, PHOSPHOR AND


OF THE LACK OF ANIMAL PROTEIN WITHIN BROILER
FOOD UPON SOME SANGUINE AND
BIOPRODUCTIVE PARAMETERS
EFECTUL NIVELULUI CALCIULUI, FOSFORULUI I AL PRIVRII DE PROTEIN
DE ORIGINE ANIMAL DIN HRANA PUILOR DE CARNE ASUPRA
UNOR PARAMETRII BIOCHIMICI SANGVINI I BIOPRODUCTIVI

C. FALC, Gh. CIORBA, Daniela LUPESCU

Cercetrile au fost efectuate pe un efectiv de 36 pui de carne


hibridul Ross 308, repartizai n 4 loturi (lotul martor i trei loturi
experimentale), pe o durat de 42 de zile. Durata experimentului fost de 42
de zile, fiind structurat pe dou subperioade de vrst (1-21 zile i 22-42
zile). Furajarea puilor din lotul martor s-a fcut cu nutre combinat
standard, n timp ce n raia puilor din loturile experimentale s-a exclus
proteina de origine animal (fina de pete). De asemenea, puii din loturile
experimentale au beneficiat de niveluri diferite de calciu i fosfor n raie.
n urma investigaiilor paraclinice privind influena raiilor furajere
difereniate, asupra parametrilor biochimici sangvini la puii de carne,
repartizai n 4 loturi: lotul martor (furaj standard 21-1); lotul experimental
E1 (privai de carbonat de calciu); lotul experimental E2 (cu aport de
carbonat de calciu, fosfat monocalcic, fr protein de origine animal);
lotul experimental E3 (fr carbonat de calciu, fosfat monocalcic i
protein de origine animal, dar cu supliment vitaminic AD3E), s-au
conturat urmtoarele concluzii: proteinemia i calcemia au fost
semnificativ mai mari la puii din lotul martor fa de cei din loturile
experimentale; calcemia i fosforemia puilor din lotul martor a fost
semnificativ mai sczute comparativ cu cei din lotul experimental E2.
Evoluia masei corporale a confirmat o scdere n greutate a puilor din
loturile experimentale E1, E2, E3 comparativ cu pui din lotul martor.
Sporul mediu zilnic pe toat durata experimental a fost mai mare la
puii din lotul martor comparativ cu cei din loturile experimentale.

Key words: broiler, macroelements, protein, blood, biochemical parameters,


bioproductive parameters

There is a strong correlation between the nutrition-based food contribution


and the metabolism of the animal organism. The maintenance of the internal
environment homeostasis is based upon the assurance through ration of the
necessary nutritive substances (proteins, glucides, lipides) and biologically active
substances (vitamins and minerals). It is obligatory to apply the optimal
percentage between the various nutrients (proteins, energizers, cellulosis, protein,

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calcium, phosphor, sodium, potassium, etc.). The assurement of the nutritive


principles through ration must have a rhythmic and equilibrate character, in terms
of quantity, and also quality (2).
Many observations revealed that the proteic amount in poultry influences
the proteinemia, albuminemia and uricemia`s levels (4).
A protein-lacking feeding during a period of 2-3 weeks leads to the
decrease of egg production and delays development in chicken, and causes
hypoproteinemia, too.
Taking into account these aspects, in this paperwork we intend to survey
the influence of a differentiate food regime upon the sanguine and bioproductive
parameters in broiler chicken.

MATERIAL AND METHODS

Researches were performed upon a number of 36 chickens, hybrid Ross


308, distributed into four groups (the control group and the experimental groups 1,
2, 3), within a long term experimentation (42 days).
In the first experimental subperiod (1-21 days), we introduced the
combined forage (C.F.) 21-1 into chicken food, and in the second subperiod (22-
42 days) we used C.F. 21-2. Chicken were kept in multi-stage battery cages (3
levels), with surfaces of 0.60 m2, namely 0.05 m2/individual.
Chicken feeding in the control group was carried out with standard
combined forage, while regarding the experimental groups (E1, E2, E3) we
excluded the animal protein from the ration.
The differentiation between the experimental groups E1, E2, E3 was
carried out through the amounts of calcium and phosphor administrated within
food, as follows: the experimental group E1 without calcium carbonate and
monocalcic phosphate; the group E2 benefitted from double amounts of calcium
and phosphor, while in the case of the experimental group E3, due to the total
lack of calcium and phosphor, vitamins A, D3 and E were supplemented in the
drinkable water.
At the end of the experimental period, we took blood samples, and with the
help of the semiautomatic biochemical analysor VET-SCREEN we determined the
following sanguine parameters: proteinemia, albuminemia, calcemia,
phosphoremia, magnesiemia, alkaline phosphatase.
The obtained results were quantized and interpreted biostatistically (the
test Mann-Withney).

RESULTS AND DISCUSSIONS

In Table 1, we present the comparative evolution of the sanguine


biochemical parameters in broilers - the control group and the experimental group
E1, which were deprived of calcium, phosphor and animal protein in their rations.
We observe that proteins in control group had a value of 4.790.18 g/dl compared

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to those from the experimental group, which had a significantly lower value
(4.010.18 g/dl), also the albuminemia in control group had a bigger value
(2.310.06 g/dl) compared to broilers from the experimental group (2.190.09
g/dl).
Table 1
The evolution of the sanguine biochemical parameters in broilers (n=7)
and the differences` signification

Sanguine Differences Mann-


biochemical Group CV% Withney
Abs %
parameters test
M 4,790,12 6,65
Total protein g/dl +0,78* 19,45 0,0106
E1 4,010,18 11,98
M 2,310,06 7,24
Albumine g/dl +0,12 5,47 0,2248
E1 2,190,09 11,03
M 10,640,17 4,33
Calcium mg/dl +2,60** 32,33 0,0017
E1 8,040,20 6,53
M 9,000,21 6,25
Phosphor mg/dl +1,26* 16,27 0,0253
E1 7,740,48 16,30
Magnesium M 2,200,12 13,89
+0,24 12,24 0,3066
mg/dl E1 1,960,13 17,66
Alkaline M 2539,8620,50 2,13
-2026,71* 44,38 0,0253
phosphatase U/L E1 4566,57821,22 47,48
*p < 0.05; **p < 0.01

Calcemia in control group chickens had the value of 10.640.17 mg/dl,


compared to the chickens from the experimental group, where we observed a
significantly reduced concentration (8.040.20 mg/dl).
Phosphoremia in the control group was 9.000.21 mg/dl, while that
belonging to the experimental group was significantly lower (7.740.48 mg/dl)
compared to that in the control group. Calcium and phosphor requirements in
broilers depend up to a level upon the level of vitamin D in food. Ca and P
amounts in ration may decrease if we add a bigger quantity of vitamin D in food
and reversely.
Researches performed by Statov et al. (5) regarding calcium and phosphor
adsorption in chicken and young substitution poultry upon groups with a calcium
level reduced with 25% and 50% revealed the fact that the body weight decreases
when the calcium levels in food increase or decrease, without being assured
statistically. Calcemia and phosphoremia decreases, too, depending on the
increased or decreased calcium level within food.
Magnesiemia in the control group chickens was 2.200.12 mg/dl,
significantly bigger compared to the chickens in the experimental group
(1.960.13 mg/dl).

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Alkaline phosphatase in the chickens within the control group had a value
of 2539.8620.5 U/L, recording a significantly distinct increase in the
experimental group (4566.57821.224 U/L).
This increase might be due to an intense osteoblasts and osteoclasts
activity, phenomena that occurs during the bones development in young chicken,
the egg laying period in hens, in rickets and osteomalacia (4).
In Table 2, we present the evolution of the sanguine biochemical
parameters from the experimental group E2, in which ration we had added a
double Ca and P level, without animal protein.
Table 2
The evolution of the sanguine biochemical parameters in broilers (n=7)
and the differences` signification

Sanguine Differences Mann-


biochemical Group X S x CV%
abs %
Withney
parameters test
Total protein M 4,790,12 6,65
+0,23 5,04 0,1598
g/dl E2 4,560,06 3,32
M 2,310,06 7,24
Albumine g/dl +0,87** 60,41 0,0017
E2 1,440,09 16,43
Calcium M 10,640,17 4,33
-1,32** 11,03 0,0049
mg/dl E2 11,960,29 6,51
Phosphor M 9,000,21 6,25
-0,74** 7,59 0,0214
mg/dl E2 9,740,19 5,20
Magnezium M 2,200,12 13,89
+0,77** 53,84 0,0021
mg/dl E2 1,430,08 15,50
Alkaline M 2539,8620,50 2,13
phosphatase -1441 37,20 0,5653
E2 3980,0821,02 53,86
U/L
*p<0.05; **p<0.01

Analysing the information presented in the table, we observe that there is a


less significant difference between the proteinemia values in both chicken groups.
In the control group, proteinemia had a value of 4.790.12 g/dl, respectively
4.560.06 g/dl in the experimental group E2.
Albuminemia is distinctly significant between the two chicken groups. We
observe a decrease (1.440.09 g/dl) in the experimental group, while in chicken
from the control group albuminemia had a value of 2.310.06 g/dl. This decrease
might be due to the lack of amount, hepatic disorders, dehydration, etc.
Calcemia was much increased in the experimental group (11.960.17
mg/dl), compared to the control group (10.640.17 mg/dl). This increase is due to
a ration supplementation with bog amounts of calcium and phosphor, and
phosphoremia has bigger values (9.740.19 mg/dl) in the case of the experimental
group compared to the chickens from the control group (9.000.21 mg/dl).

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Magnesiemia has a much reduced concentration in the experimental group


chickens (1.430.08 mg/dl) compared to those from the control group (2.200.12
mg/dl).
Alkaline phosphatase had a more intense activity in broilers within the
experimental group (3980.0821.02 U/L) compared to those within the control
group (2539.8620.50 U/L).
In Table 3, we present the values of the sanguine biochemical parameters in
broiler chickens from the experimental group E3, deprived of calcium, phosphor
and animal protein, but with a vitamin supplement (AD3E).
Table 3
The evolution of the sanguine biochemical parameters in broilers (n=7)
and the differences` signification

Sanguine Differences Mann-


biochemical Group X S x CV%
abs %
Withney
parameters test
Total protein M 4,790,12 6,65
+0,92 23,77 0,0017
g/dl E3 3,870,14 9,40
Albumine M 2,310,06 7,24
+0,22 10,52 0,1252
g/dl E3 2,090,12 15,02
Calcium M 10,640,17 4,33
-2,34** 28,19 0,0017
mg/dl E3 8,300,14 4,45
Phosphor M 9,000,21 6,25
+1,89** 26,58 0,0021
mg/dl E3 7,110,21 7,76
Magnesium M 2,200,12 13,89
+0,3 15,78 0,0639
mg/d E3 1,900,13 18,73
Alkaline M 2539,8620,50 2,13
phosphatase +1019,52** 67,05 0,0017
E3 1520,342813,03 49,15
U/L
*p<0.05; **p<0.01

Proteinemia was significantly lower (3.780.14 g/dl) in the experimental


group compared to the control group chickens (4.790.12 g/dl).
In the case of albuminemia, too, we observe that there is a difference
between the broiler groups. In the experimental group, it was 2.090.12 g/dl
compared to chicken from the control group (2.310.06 g/dl).
Phosphoremia was distinctly significant in the case of the experimental
group chicken (7.110.21 mg/dl) compared to those from the control group
(9.000.21 mg/dl). This decrease is justified by the lack of amount.
Magnesiemia in the experimental group was lower (1.900.13 mg/dl) that
that in the control group (2200.12 mg/dl). Both values are concordant with the
references values within the literature of specialty.
Alkaline phosphatase is significantly lower in the case of the chicken from
the experimental group E3 (1520.34283.03 U/L) compared to those from the
control group (2539.8620.50 U/L).

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Ciulan (1), carrying out researches regarding the activity of alkaline


phosphatase and creatinin phosphokinase, in broilers, fed with supplements of
vitamin E and selenium, observes a distinctly significant increase of the alkaline
phosphatase activity in broilers from the groups whose food was supplemented
with vitamin E and selenium. Reversely, creatinin phosphokinase values are
concordant to the physiological ones. Supplements with organic selenium assured
a better hepatic and muscle protection through a lower activity of creatinin
phosphokinase.
In Table 4, we present data concerning the average weight and the average
daily gain for the whole experimental period.
The evolution of the body mass confirms a weight decrease of the chicken
from the experimental groups (E1, E2, E3) compared to those from the control
group (M: 2311.1 g/individual, E1: 1941.2 g/ind., E3: 2105.5 g/ind.). The average
daily gain, during the whole experimental period, was bigger in the case of the
control group, compared to the experimental ones.
Table 4
Body weight and average daily gain in broiler during the experimental period

Specification Control E1 E2 E3
Average weight (g) at the age of 1 day 45 45 45 45
Average weight (g) at the age of 14 days 166,6 153,3 164,4 147,7
Average gain (g) at the age of 14 days 8,68 7,73 8,52 7,33
Average weight (g) at the age of 28 days 755,5 650,5 660,3 680,6
Average gain (g) at the age of 28 days 25,37 21,62 21,97 22,7
Average weight (g) at the age of 42 days 2311,1 1941,2 2021,1 2105,5
Average gain (g) at the age of 42 days 53,95 45,14 47,05 49,05

CONCLUSIONS

Total proteins (+19.45%) and sanguine calcium (32.33%) are significantly


higher (p<0.05) in the control group compared to those in the experimental group
E1, and phosphor (-7.59%) and alkaline phosphatase (-44.38%) are significantly
lower in the control group, compared to the experimental group E1.
In the case of albuminemia (+5.47%) and magnesiemia (+12.24%), we do
not observe significant differences (p>0.05) between the control group and the
experimental group E1.
Albuminemia (+60.41%) and magnesiemia (+53.84%) have significantly
higher concentrations (p<0.01) in the chickens from the control group compared
to those from the experimental group E2, and calcemia (-11.03%) and
phosphoremia (-7.59) are significantly lower (p<0.01) in the control group
compared to the chicken from the experimental group E2.
We didnt observe any significant differences (p<0.01) between the broiler
within the control group and those from the experimental group E2 concerning
proteinemia (+5.04%) and alkaline phosphatase (-37.20%).

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Total protein (+23.77%), calcemia (+28.19), phosphoremia (26.58%) and


the alkaline phosphatase (+67.05%) have significantly higher values (p<0.01) in
the control group compared to the experimental group E3.
We didnt record any significant differences (p>0.05) between the chicken
within the control group and those from the experimental group E3 concerning
magnesiemia (+15.78%).
The evolution of the body mass confirms a weight decrease in the case of
the broiler within the experimental groups E1, E2, E3 compared to the broiler
within the control group (M: 2311.1 g/individual, E1: 1941.2 g/individual, E2:
2021.1 g/individual, E3: 2105.5 g/individual). The average daily gain during the
whole experimental period was higher in the broiler within the control group
compared to those from the experimental groups.

BIBLIOGRAPHY

1. Ciulan, L.V., 2004 Cercetri privind activitatea fosfatazei alcaline i a


creatinfosfokinazei la puii de carne hrnii cu adaosuri de vitamina E i seleniu,
Lucr. t. de Med. Med. Vet. Timioara, XXXVII, 250-252.
2. Criniceanu, E., Olimpia Colibar, Falc, C., 1994 Evoluia indicilor productivi la
puii de carne hrnii difereniat, Lucr. t. de Med. Vet. Timioara, XXVII, 65-69.
3. Falc, C., Ciorba, Gh., 1997 Semiologie veterinar practic, Ed. Eurobit,
Timioara.
4. Prvu, Gh., 1992 Supravegherea nutriional metabolic a animalelor, Ed. Ceres,
Bucureti.
5. Statov, C., Ognean, L., Mihai, Gh., Pusta, D., Statov, D., Gondo Maria, 1995
Aspecte privind interrelaia dintre diferite nivele de calciu i vitamina D3 n
alimentaia ginilor outoare, Buletin U.S.A.M.V. Bucureti, 49, 149-155.
6. Stilborn, H., Waldroup, P.W., 1990 Production of broiler chicken on low protein
diets, utilising commercially avaiable amino-acids, Poultry Science, 69, 1207-
1211.

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ON THE BIOCOMPATIBILITY OF POLY (URETHANEUREA)


DOPED WITH SILVER NANOPARTICLES
OBSERVAII PRIVIND BIOCOMPATIBILITATEA UNUI POLIURETANUREE
DOPAT CU NANOPARTICULE DE ARGINT

B.A. HAGIU, L.C. BURTAN


U.S.A.M.V. Iai
Maria Sultana MIHAILOVICI
U.M.F. Iai
V. URA
Univ. Al. I. Cuza Iai
C. CIOBAN
ICMPP Petru Poni Iai
FERARIU D.
Spitalul Sf. Spiridon Iai

The biocompatibility study of Silver nanoparticles doped


poly(urethaneurea) by subcutaneous implants on rabbits revealed that
these materials are very well accepted, the inflammatory reaction being
extremely weak. By contrast to the poly(lactate urethanes) previously
investigated, the polyurethanes doped with Silver nanoparticles do not
adhere to tissues, and promote very fast high quality skin regeneration,
even of hair follicles.

Key words: silver, poly(urethaneurea), skin, regeneration

The polyurethanes are high biocompatible materials (1, 2, 4, 5, 6, 7, 8).


Previous works of our team were revealed that fibroblasts adhered on
poly(lactate)urethane film surface treated with Ar ion beams of various energies,
and the implant stimulates proliferation of the collagen material. These properties
enabled the poly(lactate)urethane films to adhere to the derma. (3, 9). The pores
existing in the polyurethane film act as traps for antigens and inside them
polymorphonuclear neutrophyles, fibroblasts and macrophages accumulate. The
macrophages secrete growth factors that create a neoformation vascular network
which expands on the entire film thickness. In the tissue near the implant, a
moderate inflammatory reaction was observed.
In the present work we evaluated the biocompatibility of poly
(urethaneurea) doped with silver nanoparticles, by analyzing the local reactions at
skin level.

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MATERIALS AND METHODS

The investigations were performed on 6 adult rabbits on which 2


subcutaneous implants (1 x 1 cm2) were performed in the lombosacral region.
One of the implant was the simple poly (urethaneurea) and the other was the
doped material with 20 ppm silver nanoparticles (2-10 nm diameters). The
implants were done observing the animal protection rules (Novocain anesthesia).
After two weeks, under local anesthesia, the implants were extracted together
with the tissue next to the postoperatory cicatrices. The tissue and the poly
(urethaneurea)s samples were first preserved in neutral formol (15%).
Subsequently, the samples were included in paraffin and cut in slices of 4 m
thickness using a microtome, cutting being performed perpendicularly on the
implant and skin surfaces. The slices were colored by hematoxylin-and-eosin
method.

RESULTS AND DISCUSSIONS

By contrast to the poly(lactate)urethane that strongly adhered to the tissue


(3), tissue adhesion was not observed in case of the polyurethane doped with 20
ppm silver nanoparticles (2-10 nm diameters). But, the silver doped poly
(urethaneurea) determined a fast cicatrisation of the operatory plague,
phenomenon that was not observed for poly(lactate)urethane or simple
polyurethane.
In case of the simple polyurethane implant, the microscopically
investigation revealed an inflammatory reaction of moderate intensity at skin
level, and a moderate quantity of inflammatory exudation was observed (fig. 1).

Fig. 1. - Moderate inflammatory reaction at skin level.


Histological section (H.E. ob. 20x).

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The investigated samples show a small quantity of inflammatory exudates


present inside the poly (urethaneurea) pores. On the implant surface, fibroblasts
adhesion and collagen synthesis was observed (fig. 2).

Fig. 2 - Transversal cut through a poly (urethaneurea) film implant (H.E. - Ob.10x).

Fig.3. - Low inflammatory reaction at skin level. Good regeneration, even for hair follicles.
Histological section (H.E. ob. 4x).

The cutaneous tissue prelevated from the cicatriceal area around the
implant of polyurethane doped with silver nanoparticles showed some interesting
aspects during the photonic microscopy investigations. Besides the low
inflammatory reaction (small quantity of exudates), a very good epidermal
regeneration, also observed in some treatments with ionic silver, but at higher

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concentrations (10), and a low derma proliferation were observed. The most
interesting phenomenon observed was the regeneration of hair follicles (the hair
follicles from regeneration area were compared to those from surrounding areas).
A very important fact observed in our study was that in case of simple
polyurethane implants, the skin regenerated without hair follicles (fig. 1, 3, 4).

Fig. 4. - Hair follicles of normal aspect. Histological section (H.E. ob. 10x).

The transversal sections of the Silver doped poly (urethaneurea) implants


showed fibroblast adhesion and collagen synthesis on the surface. Inside pores,
the inflammatory exudate is almost absent and there are no polymorphonuclear
neutrophyles or macrophages, which are arguments for the very low inflammatory
reaction observed (fig. 5).

Fig. 5. - Transversal section through poly (urethaneurea) (H.E. Ob. 4x).

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One can conclude that doping the poly (urethaneurea) with silver
nanoparticles give rise to some important properties that promote a very rapid
cicatrisation and, probably, makes impossible the tissue adhesion.

CONCLUSIONS

An implant of polyurethane doped with Silver nanoparticles determines a very


low inflammatory reaction.
By contrast to the polyurethanes previously investigated, the doped material
investigated in the present work did not adhere to tissue.
An important feature of the Silver doped polyurethane is the favorization of rapid
and good quality skin regeneration, which could be of interest for the hair
follicles, also.

REFERENCES

1. Anderson, A.J., 1998 - Biocompatibility of poly(etherurethane urea) containing


dehydroepiandrosterone, J. Biomed. Mater. Res., Aug;41(2):192-201.
2. Gutowska, A., Bae, Y.H., Jacobs, H., Feijen, J., Kim, S.W., 1995 - Heparin release
from thermosensitive polymer coating: In vivo studies, Journal of Biomedical
Materials Research, 29: 811.
3. Hagiu, B.A., Fntnaru, M., Burtan, L.C., Maria Sultana Mihailovici, Elena-Doina
Murarescu, Melnig, V., ura, V., 2003 - In Vivo Intradermic Study of Cell
Adhesion to Ion-Implanted Polyurethanes, Lucrri tiinifice, vol 46, seria
Medicin Veterinar, pag 83-88.
4. Han, H.S., et al., 2001 - Studies of a novel human thrombomodulin immobilized
substrate: surface characterization and anticoagulation activity evaluation,
Journal of Biomaterials Science Polymer Edition Vol. 12, No. 10:1075-1089.
5. Jayabalan, M., Rathinam, K., Kumary, T.V., Mohanty, M., 1989 - In vivo
biocompatibility o aliphatic segmented polyurethane in rabbit, Journal of
Biosciences. Sep, 14(3): 289-99.
6. Kwon, I. C., Bae, Y.H., Kim, S.W., 1994 - Heparin release from polymer complex,
Journal of Controlled Release 30: 155.
7. Lamba, N.M.K., Woodhouse, K.A., Cooper, S.L., 1998 - Urethanes in biomedical
applications, Boston, CRC Press.
8. Park, K.D., Kim, W.G.; Jacobs, H., Okano, T., Kim, S.W., 1992 - Blood
compatibility of SPUU-PEO-heparin grafted copolymers, Journal of Biomedical
Material Research, 26: 739.
9. ura, V., Apostu, M.O., Melnig, V., Ciobanu, C., Hagiu, B.A., 2003 Preparation
and characterization of biocompatible polylactaturetane, Timioara Medical
Journal, Vol. 53, Suppl. 2, pag. 127-130.
10. Konrad, D. Tsunoda, M. Weber, K. Corney, S.J. Ullmann, L., 2002 Effects of a
topical silver sulfadiazine polyurethane dressing (Mikacure) on wound healing in
experimentally infected wounds in the pig. A pilot study, Journal of experimental
animal science, Vol.42, Part 1, pag. 31-43.

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CERCETRI PRIVIND LEUCOCITEMIA, EOZINOCITEMIA


I BAZOFILECITEMIA LA IEPURELE DOMESTIC
RESEARCHES REGARDING LEUCOCYTEMY, EOSINOCYTEMY AND
BASOPHILCYTEMY AT DOMESTIC RABBIT

Roxana LAZR, M. LAZR


U.S.A.M.V. Iai

Researches were made upon leucocytemy, eosinocytemy,


basophilcytemy on a number of 30 rabbits belonging to the races New
Zeeland White, Butterfly and Chinchila, raised in small husbandries,
distributed in 5 groups depending upon their age, sex and physiological
state.
The results obtained showed the fact that the number of total
circulatory leucocytes increased proportionally with age until 1 year old
after which they lightly decreased maintaining constant until adult age.
There were no variations depending on sex, at pregnant females the
leucocytemy being more raised. Eosinophiles showed increases
proportional with the age and didnt show great variations depending on
sex and physiological state. The basophiles also increased proportionally
with the age, sex and physiological state, at males and pregnant females
being registered bigger values.

Examenul hematologic prezint o deosebit importan n evaluarea


integritii nutriional metabolice a efectivelor de animale i o semnificaie
major n aprecierea strii de sntate i reactivitate specific natural a
organismului (componenta leucocitar a sngelui).
Necesitatea evalurii integritii nutriional metabolice a efectivelor de
animale rezid din faptul c multe carene nutriionale pot produce boli tot att de
grave sau uneori chiar mai grave, dect bolile produse de germeni patogeni, iar
pagubele produse de tulburrile nutriionale sunt mai mari ntruct nregistreaz
adeseori o evoluie subclinic.
O importan deosebit o prezint alimentaia femelelor gestante, acestea
avnd apetitul i asimilaia mrite, asigurnd necesarul de substane nutritive i
pentru ft. Creterea necesarului de protide, glucide, minerale, vitamine (A, B, D,
E) este asigurat de ficatul gestantei care i intensific funciile. n ficatul i
sngele gestantelor scade glicogenul i crete glucoza, colesterolul, srurile
minerale (Ca, P) i cantitatea de aminoacizi, care sunt transportate spre placent
cu prioritate.
Aceste considerente justific utilizarea testelor de profil metabolic i n
investigarea strilor de boal. Datele obinute n acest mod arat strile de
dezechilibru homeostatic al organismului i servesc totodat la aplicarea celor mai
adecvate msuri de prevenire a agravrii lor i redresarea echilibrului.
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MATERIAL I METODE

Experimentul a fost efectuat pe un numr de 30 de iepuri metii din rasele


Alb Neozeelandez, Fluture i Chinchila, crescui n sistem gospodresc. Conform
protocolului experimental, animalele au fost distribuite cte 6 capete pe 5 loturi
dup cum urmeaz:
- Lot 1 (n=6 ) - iepuri tineri n vrst de 4-6 luni masculi i femele;
- Lot 2 (n=6) - iepuri n vrst de 6 12 luni, masculi i femele;
- Lot 3 (n=6) - femele n vrst de 12 luni negestante;
- Lot 4 (n=6) - femele n vrst de 12 luni gestante;
- Lot 5 (n=6) - masculi n vrst de 12 luni.
Pentru efectuarea determinrilor cantitative i calitative ale elementelor
figurate sanguine s-au utilizat metodele clasice (numrare n camer Burker-
Trk).
Rezultatele obinute au fost prelucrate statistic, determinndu-se valorile
aritmetice medii i deviaia standard.

REZULTATE I DISCUII

n tabelele 1-5 sunt prezentate valorile medii i deviaia standard a


numrului de leucocite, eozinofile i bazofile, n funcie de vrst, stare
fiziologic i sex.
Tabelul 1
Variaiile individuale i valorile medii ale leucocitemiei, eozinofilelor i bazofilelor
la iepurii n vrst de 4-6 luni (L1)

Nr. Leucocite Eozinofile Bazofile


Crt. Nr./mm % nr./mm % nr./mm
1 4500 2 90 2 90
2 4750 0 0 3 142,5
3 5350 1 53,5 4 214
4 6950 1 69,5 5 347,5
5 7950 2 159 4 318
6 13850 1 158,5 3 415,5
X 7225,00 1,17 88,42 3,50 254,58
sX 1432,47 0,31 25,36 0,43 51,59
CV % 48,56 64,52 70,25 29,97 49,64
X = media aritmetic;
s X = abaterea standard a mediei;
CV % = coeficientul de variabilitate

Valorile medii ale leucocitemiei (nr./mm) la lotul alctuit din 6 iepuri n


vrst de 4-6 luni a fost de 7225 1434,35. Limitele de variaie au fost cuprinse
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ntre 4500 i 13850. Media bazofilemiei este de 254,58 bazofile/mm snge,


reprezentnd 3,5% din numrul total de leucocite circulante. Limitele de variaie
au fost cuprinse ntre 90 i 415,5 bazofile/mm snge (2-5%).
Din analiza datelor tabelului 1 se constat c media eozinofilemiei la iepurii
n vrst de 4-6 luni a fost de 84,41 eozinofile/mmsg, reprezentnd 1,16% din
numrul total de leucocite circulante. Limitele de variaie au fost cuprinse ntre
53,5 i 159 eozinofile/mm de snge (1-2%).
La lotul de iepuri cu vrsta 6-12 luni (tabelul 2) s-a constatat c valorile
medii ale numrului de leucocite (nr./mm) au fost de 10700 936,20. Limitele
de variaie au fost cuprinse ntre 7100 i 13350. n ceea ce privete media
bazofilemiei aceasta a fost de 863,16 bazofile/ mm snge, reprezentnd 10% din
numrul total de leucocite circulante. Limitele de variaie au fost cuprinse ntre
336 i 1202,5 bazofile/mmde snge (5-16%).
Tabelul 2
Variaiile individuale i valorile medii ale leucocitemiei, eozinofilelor i bazofilelor la
iepurii n vrst de 6-12 luni (L2)

Nr. Leucocite Eozinofile Bazofile


Crt. Nr./mm % nr./mm % nr./mm
1 7100 2 42 16 336
2 9250 1 92,5 13 1202,5
3 10500 1 105 10 1050
4 11400 1 114 9 1026
5 12600 1 126 5 650
6 13350 2 267 7 934,5
X 10700,00 1,33 124,42 10,00 866,50
sX 934,97 0,21 30,89 1,63 129,73
CV % 21,40 38,73 60,82 40,00 36,67

Media eozinofiliei a fost de 124,41 eozinofile/mm snge, reprezentnd


1,33% din numrul total de leucocite circulante, limitele de variaie fiind cuprinse
ntre 42 i 267 eozinofile/mm de snge (1-2%).
Variaii medii ale leucocitemiei (nr./mm) la lotul alctuit din 6 femele n
vrst de peste 1 an au fost de 9083,35 194,14. Limitele de variaie au fost
cuprinse ntre 7600 i 10700. Media eozinofiliei la femelele adulte a fost de
140,66 eozinofile/mm de snge, reprezentnd 1,5% din numrul total de
leucocite circulante. Limitele de variaie au fost cuprinse ntre 76 i 214
eozinofile/mm de snge (1-2%).
Media bazofilemiei la femelele adulte a fost de 774,5 bazofile/mm de
snge, reprezentnd 8,33% din numrul total de leucocite circulante. Limitele de
variaie au fost cuprinse ntre 380 i 1070 bazofile/mm de snge (5-10%).

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Tabelul 3
Variaiile individuale i valorile medii ale leucocitemiei, eozinofilelor i bazofilelor
la femelele de 12 luni negestante (L3)

Nr. Leucocite Eozinofile Bazofile


Crt. Nr./mm % nr./mm % nr./mm
1 7600 1 76 5 380
2 7900 1 79 7 553
3 9100 1 91 9 819
4 9500 2 190 9 855
5 9700 2 194 10 970
6 10700 2 214 10 1070
X 9083,33 1,50 140,67 8,33 774,50
sX 474,99 0,22 26,53 0,80 106,25
CV % 12,81 36,51 46,19 23,60 33,60

Tabelul 4
Variaiile individuale i valorile medii ale leucocitemiei, eozinofilelor i bazofilelor la
femelele n vrst de 12 luni gestante (L4)

Nr. Leucocite Eozinofile Bazofile


Crt. Nr./mm % nr./mm % nr./mm
1 8100 2 162 6 486
2 9310 1 93,1 11 1024,1
3 9250 1 146,5 9 1318,5
4 10500 1 105 7 735
5 12750 2 255 15 1612,5
6 14650 1 107,6 17 1829,2
X 10760,00 1,33 144,87 10,83 1167,55
sX 1010,25 0,21 24,55 1,80 210,61
CV % 23,00 38,73 41,51 40,62 44,19

Valorile medii ale leucocitemiei ( nr./mm) la lotul de 6 femele gestante au


fost de 10760 1011,58. Limitele de variaie au fost cuprinse ntre 8100 i 14650
dintre care 4 iepuri au avut valori mai mici dect valoarea medie (8100, 9250,
9310, 10500) i ceilali 2 iepuri au avut valori peste valoarea medie (12750;
14650).
n urma analizei datelor prezentate n tabelul 4 se constat c media
eozinofilemiei la femelele gestante este de 144,86 eozinofile/mm snge,
reprezentnd 1,33% din numrul total de leucocite circulante. Limitele de variaie
au fost cuprinse ntre 93,1 i 255 eozinofile/mm de snge (1-2%).

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Media bazofilemiei la femelele gestante este de 1167,55 bazofile/mm


snge; reprezentnd 10,5% din numrul total de leucocite circulante. Limitele de
variaie au fost cuprinse ntre 486 i 1829,2 bazofile/mm de snge ( 6-17%).

Tabelul 5
Variaiile individuale i valorile medii ale leucocitemiei, eozinofilelor i bazofilelor
la masculii n vrst de 12 luni (L5)

Leucocite Eozinofile Bazofile


Nr crt.
Nr./mm % nr./mm % nr./mm
1 6650 1 66,5 17 1130,5
2 8050 0 0 10 805
3 9120 3 273,6 18 1641,6
4 9720 2 194,4 12 1166,4
5 9720 1 97,2 16 1555,2
6 10900 2 219 13 1423,5
X 9026,67 1,50 141,78 14,33 1287,03
sX 607,78 0,43 42,37 1,28 127,50
CV % 16,49 69,92 73,20 21,91 24,27

Lotul alctuit din 6 masculi aduli a nregistrat valori medii ale


leucocitemiei (nr./mm) de 9360 630,78. Limitele de variaie au fost cuprinse
ntre 6650 i 10950. Media eozinofilemiei la masculii aduli este de 141,78
eozinofile/mm snge, reprezentnd 1,5% din numrul total de leucocite
circulante. Limitele de variaie au fost cuprinse ntre 66,5 i 273,6 eozinofile/mm
de snge (0-3%).
Media bazofilemiei a fost de 1287,03 bazofile/mm snge, reprezentnd
14,33% din numrul total de leucocite circulante. Limitele de variaie au fost
cuprinse ntre 805 i 1641,6 bazofile/mm de snge(10-18%).
Din analiza general i prelucrarea matematic a rezultat c numrul de
bazofile a variat nesemnificativ ntre loturile L1, L 3 i foarte semnificativ ntre
loturile L1, L2, L5 i ntre L3,L5 .
Leucocitele pot prezenta variaii foarte mari (4000- 14000) i sunt n numr
mai mic la iepurii tineri. La vrsta de peste un an majoritatea autorilor arat o
cretere a numrului de leucocite cu peste 2000/mm fa de tineretul sub 6 luni.
Bazofilele la iepuri se deosebesc prin numr i aspect morfologic de
bazofile de la alte specii de mamifere. Iepurele este singurul animal domestic la
care numrul de bazofile este crescut, cu o valoare medie de 3,8-12,5% dup
Marcu (1993) ;2,4-31% dup Nemi (1986); 6,5-30% dup Scherner (1967).

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Tabelul 6
Semnificaia statistic a diferenelor dintre mediile caracterelor analizate la cele
5 loturi experimentale (L1L5)

Semnificaia statistic (Tukey) a diferenelor dintre medii pentru


Loturile caracterele:
comparate Leucocite Eozinofile Bazofile
nr/mm % nr./mm % nr./mm
L1 X L2 n.s. n.s. n.s. semnif. pt. w1 (0,05) semnif. pt. w1 (0,05)
w (6,50)>w1 (5,35) w (611,92)>w1
(559.05)
L1 X L3 n.s. n.s. n.s. n.s. n.s.
L1 X L4 n.s. n.s. n.s. semnif. pt. w2 (0,01) semnif. pt. w2 (0,01)
w (7,33)>w2 (6,61) w (912,97)>w2
(690,87)
L1 X L5 n.s. n.s. n.s. semnif. pt. w2 (0,01) semnif. pt. w2 (0,01)
w (10,83)>w2 (6,61) w (1032,45)>w2
(690,87)
L2 X L3 n.s. n.s. n.s. n.s. n.s.
L2 X L4 n.s. n.s. n.s. n.s. n.s.
L2 X L5 n.s. n.s. n.s. n.s. n.s.
L3 X L4 n.s. n.s. n.s. n.s. n.s.
L3 X L5 n.s. n.s. n.s. semnif. pt. w1 (0,05) n.s.
w (6,00)>w1 (5,35)
L4 X L5 n.s. n.s. n.s. n.s. n.s.
Testul n.s. n.s. n.s. *** ***
Fisher F > F 0,001 (4,25 F > F 0,001 (4,25
G.L.) G.L.)
9,3675 > 6,4900 8,8943 > 6,4900
W = diferena dintre mediile loturilor comparate
W1 = valoarea tabelar pentru pragul 0,05
W2 = valoarea tabelar pentru pragul 0,01
*** = valori foarte semnificative pentru testul Fisher
n.s. = diferene nesemnificativ.

CONCLUZII

Analiza rezultatelor obinute la iepurele domestic (Oryctologus Cuniculus),


crescut n sistem gospodresc, a scos n eviden c indicii seriei albe studiai
(numrul de leucocite, eozinofilemia i bazofilemia) prezint variaii legate de
vrst, sex i stare fiziologic astfel:
1. Numrul de leucocite totale circulante a crescut proporional cu vrsta
pn la un an dup care au sczut uor meninndu-se constante la vrsta adult.
Nu s-au nregistrat variaii n funcie de sex. La femelele gestante leucocitemia a
fost mai crescut.

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2. Eozinofilele au nregistrat creteri proporionale cu vrsta i nu au


prezentat variaii mari n funcie de sex i stare fiziologic .
3. Bazofilele au crescut de asemenea proporional cu vrsta i n funcie de
sex i stare fiziologic, la masculi i femele gestante nregistrndu-se valori mai
ridicate.

BIBLIOGRAFIE

1. Ames A, Li YY., Heher E.C., Kimble C.R. Energy metabolism of rabbit


retina as related to function: high cost of Na transport
2. Argente M.J, Santacreu M.A., Climent A., Blasco A Effecs of intrauterine
crowding on uterine available space per fetus in rabbits,Valencia, Spain
3. Ciudin Elena - Biologia animalelor de laborator, Editura Alfa, Iai 2004
4. Blasco A., Ortega A.J., Santacreu A.M., Climent A. - Divergent selection
for uterine capacity in rabbits. Genetic parameters and response to
selection
5. Marcu Elena, Condrea M., Costachescu Elena, Hagiu N., Pavel Geta -
Aspectul profilului hematologic la iepurele domestic. Lucrare tiinific,
Universitatea Agronomic Iai, vol. 35-36, 1992-1993
6. Marcu Elena, Costachescu Elena, Condrea M., Hagiu N., Pavel Geta -
Corelaie privind interrelaiile fenotipice principale, nsuiri productive i
unii indici ai profilului metabolic la iepurele domestic. Lucrare tiinific,
Universitatea Agronomic Iai, vol. 35-36, 1992-1993.
7. Moise KJ Jr. Saade G., Knudsen L. - Ultrasound-guided cardiac blood
sampling of the rabbit fetus. Departament of Obstetrics and Gynecology,
Baylor College of Medicine, Houston, Tex.
8. Moce M.L., Santacreu M.A., Climent A., Peiro R. - In vivo development of vitrified
rabbit embryos: effects on prenatal survival. Preliminary results. Departamento
de Ciencia Animal, Universidad Politecnica de Valencia, Spain
9. Michael L. Jennings - Volume-sensitive K+/Cl cotransport in rabbit Erytrocites:
Analysis of the rate - limiting activation and inactivation events - The Journal of
General Psyiology, vol 114, number 6, December 1, 1999
10. Navaro J.A., Sottovia-Filho D., Taga R., Toledo-Filho J.L. - Histological study on
the prenatal development of the cheek-teeth of rabbits. Arch Histol Jpn. 1978
Jun;41(3)
11. Vanessa K. Lester, DVM; Heather L. Tarpley, DVM; Kenneth S. Latimer, DVM -
Small Mammal hematology: Leucocyte identification in rabbits. College
Veterinary Medicine, University of Georgia, Athens GA 30602-7388.
12. Sandi Koi - Domestic Baby Bunnies and Their Mom
13. *** - Histological different from neutrophil
14. *** - Domestic Rabbit
15. *** - Biochemistry Reference Values
16. *** - Bun or Blood Urea Nitrogen in rabbits
17. *** - Lowering Blood Calcium
18. *** - Embryonic development of the rabbit pineal gland
19. www.pubmed.gov
20. www.Kindplanet.org
21. www.Medirabbit.com

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APRECIERI ASUPRA SEMNIFICAIEI


MORFOFUNCIONALE A COMPLEXELOR
NEUROINSULARE
APPRECIATIONS ABOUT THE MORPHO FUNCTIONAL SEMNIFICATION
OF THE NEURO ISLET COMPLEXES

V. MICLU, A. DAMIAN, L. OANA,


C. LISOVSCHI, F. STAN

Histological examination was performed on pancreas samples


collected from two male nutria. We observed the presence of the neuro islet
complexes, alongside the Langherans isles with classical structure. The
placement and the aspects of the endocrine cells and of the vegetative
neurons in these neuro isle complexes do not suggest a significant morpho-
functional change; possibly that these complexes appeared during
embryogenesis.

Key words: islet of Langherans; pancreas; vegetative neurons

INTRODUCERE

Inervaia insulelor endocrine pancreatice a fcut obiectul a numeroase


cercetri, care au evideniat faptul c ele sunt deservite de o inervaie foarte
bogat, att simpatic ct i parasimpatic (Samols, 1991; Hornet, 1992). Cu toate
informaiile acumulate de-a lungul anilor, inervaia insulelor pancreatice nu este
pe deplin lmurit nici la ora actual. Ca o curiozitate a aprut semnalarea n
imediata vecintate a insulelor pancreatice a unor ganglioni vegetativi. Mai mult,
unii cercettori au remarcat faptul c uneori pot fi observate celule nervoase
vegetative, chiar n interiorul insulelor pancreatice (Capenhaut, 1927; Simard,
1942, Fujita, 1969; Miclu, i col. 2003). Totui, numrul cercettorilor care au
semnalat astfel de formaiuni de-a lungul timpului, este destul de mic, iar
semnificaia morfofuncional a lor a rmas nc nelmurit.

MATERIAL I METODE

Studiul a fost realizat pe doi masculi de nutrie n vrst de 8, respectiv 10


luni, clinic sntoi i cu stare de ntreinere foarte bun. Au fost recoltate
fragmente de pancreas , sub form de felii cu grosimea de 3-4 mm i fixate n
amestec Stieve 24 de ore. Dup includere n parafin s-au practicat seciuni cu
grosime de 5 micrometrii, care au fost colorate cu metoda tricromic Masson,
modificat de Goldner.

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REZULTATE I DISCUII

Examinarea microscopic a seciunilor de pancreas a evideniat c nu exist


diferene structurale semnificative ntre componenta endocrin a pancreasului de
nutrie i cea de la alte mamifere. i n cazul nutriei, componenta endocrin este
reprezentat de insule Langerhans de diferite forme i mrimi, rspndite
neuniform printre acinii pancreatici. Pe lng insulele Langerhans cu structur
clasic, au fost evideniate complexe neuroinsulare, n care alturi de celule
endocrine existau i neuroni vegetativi. Numeric, insulele Langerhans predominau
net, dar i complexele neuroinsulare erau relativ bine reprezentate. Complexele
neuroinsulare apar bine conturate printr-o capsul conjunctiv discret i cu
structur lax. Ele sunt foarte diferite ca mrime, de la complexe foarte mari pn
la formaiuni alctuite din cteva celule. Ca form ele sunt de asemenea
polimorfe, existnd complexe neuroinsulare rotunde, ovale, alungite, foarte
alungite, septate. Raportul ntre componenta endocrin i cea nervoas este foarte
diferit de la o formaiune la alta. n unele complexe celulele endocrine sunt
majoritare (fig. 1), n altele predomin celulele nervoase (fig. 2). ntre cele dou
extreme, exist foarte multe situaii intermediare, dar indiferent care component
predomin, ele nu apar intercalate ci fiecare ocup o parte a complexului
neuroinsular. Aspectul este uor de sesizat chiar i n cazul complexelor n care
una dintre componente este net majoritar, cealalt chiar dac conine doar cteva
celule, ele apar dispuse grupat la periferia formaiunii. Aceste aspecte sugereaz
c inervaia celulelor endocrine nu necesit neaprat contact intim ntre ele i
neuroni. Din acest punct de vedere, prezena neuronilor n interiorul complexelor
neuroinsulare nu pare neaprat un avantaj n comparaie cu inervarea insulelor
Langerhans clasice, care este asigurat de neuroni de la nivelul ganglionilor
vegetativi aflai la distan mai mare sau mai mic de insule. Ganglioni vegetativi
au fost evideniai, fie la distan de insulele Langerhans fie n apropierea
acestora. Unii ganglioni apar dispui n contact att de intim cu insulele nct pe o
anumit poriune, capsula pare comun celor dou formaiuni. Dar relaia intim
dintre cele dou formaiuni pare mai mult de vecintate i nu o necesitate
morfofuncional. De altfel, insulele Langerhans aflate n apropierea ganglionilor
vegetativi sunt comparabile din toate punctele de vedere cu cele aflate la distan.
Asta sugereaz c inervaia celulelor endocrine pancreatice se poate face eficient
de neuroni aflai la distan de insulele Langerhans, prezena ganglionilor
vegetativi n apropierea acestora pare ntmpltoare i dup toate probabilitile
nu asigur o inervaie mai eficient dect cei aflai la distan.

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Fig. 1 - Complex neuroinsular n care celulele endocrine sunt majoritare


(Col. Tricromic Goldner, 500X).

Fig. 2 - Complex neuroinsular n care neuronii sunt majoritari


(Col. Tricromic Goldner, 500X).

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Celulele endocrine din componena complexelor neuroinsulare apar


comparabile, din toate punctele de vedere cu cele din insulele Langerhans. Nici
neuronii vegetativi din complexe nu sunt cu nimic diferii fa de cei din
ganglionii vegetativi. Aceste aspecte, alturi de polimorfismul att de accentuat al
complexelor neuroinsulare, cu raporturi i proporii att de diferite ntre
componenta endocrin i cea exocrin, ne fac s credem c dispunerea n
complexe neuroinsulare nu pare a avea o semnificaie morfofuncional, cel puin
nu una sesizabil prin examen microscopic. Se pare c prezena complexelor
neuroinsulare alturi de insulele Langerhans n pancreasul de nutrie, nu este o
necesitate morfofuncional, ci ele pot apare conjunctural. Se pare c ele, nu
realizeaz neaprat o inervaie mai eficient a celulelor endocrine, comparativ cu
cea asigurat de neuronii din ganglionii vegetativi. Cea mai plauzibil explicaie a
prezenei complexelor neuroinsulare, ar fi formarea lor n cursul embriogenezei.
Dac este aa, atunci se pare c celulele endocrine pancreatice i neuronii
vegetativi provin din aceeai foi embrionar. Sporadic au mai fost fcute
afirmaii (Buligescu i col. 1982) conform crora celulele insulare provin din
crestele neurale i nu din endoderm, cum susin cei mai muli autori (Adlersberg
i col. 1955; Diculescu i col. 1971; Bareliuc i Neagu, 1987; Raica i col. 2004).

CONCLUZII

1. Examenul histologic al pancreasului de nutrie a evideniat insule


Langerhans de form, mrime i distribuie, comparabile cu cele de la alte
mamifere.
2. Pe lng insule pancreatice cu structur clasic, au fost evideniate
complexe neuroinsulare diferite ca mrime i raport ntre componenta endocrin
i cea nervoas.
3. Distribuia celulelor endocrine i a neuronilor vegetativi n cadrul
complexelor este variabil, astfel c ea nu sugereaz un aranjament ce ar putea
avea vreo semnificaie funcional.
4. Polimorfismul accentuat i gruparea caracteristic a celulelor endocrine
i a neuronilor vegetativi din complexele neuroinsulare, ne fac s credem c ele
nu sunt o necesitate morfofuncional ci , probabil, au luat natere n cursul
embriogenezei.

BIBLIOGRAFIE

1. Adlersberg, L., S. Brtianu, C. Crian, M. Gundisch, A. Hagi Paraschiv, T.


Niculescu, C. Rmniceanu, A. upa, 1955 - Histologie, Editura medical,
Bucureti.
2. Bareliuc, Lucia, Natalia Neagu, 1987 - Embriologie uman, Editura Medical,
Bucureti.
3. Buligescu, L., B. Gheorghescu, I. Puca, I. Teodorescu Exarcu, 1982 -
Fiziologia i fiziopatologia digestiei, Edit. Medical, Bucureti.

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4. Van Capenhaut, E.,1927 - Contributions a letude de lhistogenese du pancreas chez


quelques mammiferes. Les complexes sympaticos-insulaires, Arch. Biol. Liege,
37, 121-171.
5. Diculescu I., D. Onicescu, C. Rmniceanu, 1971 - Histologie, Editura didactic i
pedagogic, Bucureti.
6. Fujita, T., 1969 - Histological studies on the neuro-insular complex in the pancreas of
some mammals, Zellforsch, 50, 94-107.
7. Hornet, N.N.,1992 - Pancreasul endocrin, Editura Academiei Romne, Bucureti.
8. Miclu, V.,C. Crciun, I. Groza, 2003 - Complexe neuroinsulare n pancreasul
endocrin de nutrie, Analele Societii Naionale de Biologie Celular, 453-460,
Editura Risoprint, Cluj-Napoca.
9. Raica, M., O. Mederle, I.D. Cruntu, Alina Pntea, A.M. Chindri, 2004 - Editura
Brumar, Timioara.
10. Samols, E., 1991 - The Endocrine Pancreas, 46-47, Ed. Raven Press, New York.
11. Simard, L.C.,1942 - Le complexe neuro-insulaire du pancreas, Rev. Can. Biol.,1, 2-
49.

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STRUCTURI FOLICULARE N LOBUL INTERMEDIAR


AL HIPOFIZEI DE TAUR
THE FOLLICULAR STRUCTURES IN PARS INTERMEDIA
OF THE BULL HYPOPHYSIS

C. COTEA, O.Z. OPREAN, Carmen SOLCAN


U.S.A.M.V. Iai
I. COTEA
U.M.F.Gr. T. Popa Iai

As a research material we have used the hypophysis of 14 Holstein


bulls at of 3-8 years. The samples have been fixed in Orth, Carnoy, and
Helly, led up to paraffin and serially sectioned at 5 m. We have finally
obtained 420 serial sections stained through the methods: Novelli, PAS,
Fontana, Steedman-Mowry, Bielschowsky, Bodian and Holmes. The
intermediate lobe is well developed in the bull. It is always adjacent to the
posterior lobe of the bull hypophysis, separated from it by a discontinuous
layer of connective tissue. Follicular structures (110-360 m) containing
colloid in the lumen were found in the pars intermedia of 14 bulls
hypophysis. This follicular structures are separated by thin connective
tissue strands, they form cysts which contain colloid. The colloid is pale-
staining materials, PAS-positive and is surrounds by simple squamous or
cuboidal epithelium.The colloid consists chiefly of a glycoprotein in the
Steedman-Mowry stain. The pars intermedia of the hypophysis makes a
hormone MSH (Melanocyte Stimulating Hormone) which is responsible for
the expansion of melanocytes from the animal skin.

Key words: follicular structures, pars intermedia, bull hypophysis

Structuri foliculare au fost observate n adenohipofiza de om, cine, porc,


berbec i obolan (1, 5, 6, 7, 8, 9, 12). Pentru a extinde aceste investigaii i la alte
specii, s-a intreprins prezenta lucrare.

MATERIAL I METOD

Ca material de cercetat au servit hipofizele de la 14 tauri Holstein, n vrst


de 3-8 ani. Cele 28 de fragmente rezultate din secionarea medio-sagital a
hipofizelor au fost fixate n Carnoy i Helly, incluse n parafin i secionate la 5
m. Coloraiile folosite au fost: PAS, Papanicolau, Novelli, MH2, Fontana,
Steedman-Mowry, Bielschowsky, Bodian i Holmes.

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REZULTATE I DISCUII

n lobul intermediar al adenohipofizei la taurii luai n studiu au fost


evideniate structuri foliculare de dimensiuni variabile (110 - 360 m). Acestea
sunt structurate dintr-un epiteliu cubic de 7-9 m, cu celulele epiteliale ce conin
nuclei ovalari de 4 m (fig. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16).
Unele structuri foliculare conin n lumenul lor un coloid omogen, PAS-
pozitiv i fr vacuole de resorbie marginale. De menionat faptul c alte structuri
foliculare din lobul intermediar al adenohipofizei de taur, coloidul apare cu
vacuole de resorbie marginale, semn al unei elaborri intense de MSH. Faptul c
structurile foliculare apar frecvente n lobul intermediar al hipofizei de taur, la
care coloidul apare cu vacuole de resorbie marginale, demonstreaz elaborarea la
un nivel ridicat de MSH.

Fig. 1. Hipofiz - lobul intermediar de taur Fig. 2. Hipofiz - lobul intermediar de taur
Holstein n vrst de 4 ani. Vezicul cu o Holstein n vrst de 4 ani. Vezicul cu o
secreie n lumen . Col. MH2; x 200 secreie n lumen . Col. MH2; x 200

Fig. 3. Hipofiz - lobul intermediar de taur Fig. 4. Hipofiz - lobul intermediar de taur
Holstein n vrst de 4 ani. Trei vezicule cu Holstein n vrst de 4 ani. O vezicul cu
o secreie n lumen. Col. MH2 x 200 lumen neregulat. Col. MH2 x 400

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Fig. 5. Hipofiz - lobul intermediar de taur Fig. 6. Hipofiz - lobul intermediar de taur
Holstein n vrst de 5 ani. Vezicule cu Holstein n vrst de 5 ani. Vezicule cu
secreie n lumen. Col. MH2 x 200 secreie n lumen. Col. MH2 x 200

Fig. 7. Hipofiz - lobul intermediar de taur Fig. 8. Hipofiz - lobul intermediar de taur
Holstein n vrst de 5 ani. O vezicul cu Holstein n vrst de 5 ani. Vezicule cu
secreie n lumen. Col. MH2 x 200 secreie n lumen. Col. MH2 x 400

Fig. 9. Hipofiz - lobul intermediar de taur Fig. 10. Hipofiz - lobul intermediar de taur
Holstein n vrst de 7 ani. Vezicule cu Holstein n vrst de 7 ani. Vezicule cu
secreie n lumen. Col. MH2 x 400 secreie n lumen. Col. MH2 x 400

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Fig. 11. Hipofiz - lobul intermediar de taur Fig. 12. Hipofiz - lobul intermediar de taur
Holstein n vrst de 7 ani. O vezicul cu Holstein n vrst de 7 ani. Dou vezicule
secreie n lumen. Col. MH2 x 400 cu secreie n lumen. Col. MH2 x 400

Fig. 13. Hipofiz - lobul intermediar de taur Fig. 14. Hipofiz - lobul intermediar de taur
Holstein n vrst de 7 ani. Dou vezicule Holstein n vrst de 7 ani. Dou vezicule
cu secreie n lumen. Col. PAS x 80 cu secreie n lumen. Col. PAS x 200

Fig. 15. Hipofiz - lobul intermediar de taur Fig. 16. Hipofiz - lobul intermediar de taur
Holstein n vrst de 7 ani. Dou vezicule Holstein n vrst de 8 ani. Dou vezicule
cu secreie n lumen. Col. PAS x 400 cu secreie n lumen. Col. PAS x 400

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CONCLUZII

1. n lobul intermediar al adenohipofizei de taur au fost remarcate structuri


foliculare, care au dimensiunile variabile (110-360 m), ce sunt delimitate de un
epiteliu cubic de 7 - 9 m.
2. n lumenul unor structuri foliculare s-a remarcat un coloid omogen, PAS-
pozitiv, fr vacuole de resorbie marginale.
3. La unii tauri, coloidul din structurile foliculare apare cu vacuole de
resorbie marginale, semn al unei elaborri intense de MSH.

BIBLIOGRAFIE

1. Bergland, R.M., & Torack R.M., 1969 - An ultrastructural study of follicular cells in
the human anterior pituitary. Am. J. Pathol. 57, 273-297.
2. Cotea C. i colab., 1986 - Dinamica celulei beta-FSH din adenohipofiza suinelor
femele n proestrus din rasa Landrace n vrst de 150- 280 zile. Lucr. t. Inst.
Agr. Iai, II, Zooteh. Med. Vet., p. 75-76.
3. Cotea C. i colab., 1988 - Aspectele morfologice i histochimice ale foliculogenezei
din ovar, n corelaie cu dinamica celular din adenohipofiza la scrofiele din rasa
Landrace. Lucr. t. Inst. de Cercetare i producie pentru creterea porcilor Peri,
vol. IV, p. 201-209.
4. Cotea C., Arseni O., 1994 - Hipotalamusul la suinele femele. Lucr. t. Univ. Agr. i
Med. Vet., seria Med. Vet., vol. 37, p. 25-27.
5. Cotea C., Solcan Carmen, Arseni O.V., 1997 Structuri foliculare n lobul
intermediar al hipofizei la suinele femele din rasa Landrace. Lucr. t. Univ. t.
Agricole i Med. Vet. Iai, vol. 40, p. 20 22.
6. Cotea C., Carmen Solcan, Cotea I., 2004 - Structuri foliculare n lobul intermediar al
hipofizei la berbec. Lucr. t., vol. 47 (6), p. 26-29.
7. Horvat E., and all., 1974 - Origin, possible function and fate of follicular cells in the
anterior lobe of the human pituitary. An electron microscopic study. Am. J.
Pathol. 77, 199-212.
8. Kagayama, M., 1965 - The follicular cell in the pars distalis of the dog pituitary gland
: an electron microscope study. Endocrinol. 77, 1053- 1060.
9. Kubo M. and all., 1992 - Follicular structures in the hypophysis of pigs. Bull. Natl.
Inst. Anim. Health 98, 9-13.
10. Liwska, J., 1978 - Investigations of ultrastructure of the adenohypophysis in the
domestic pig (Suis scrofa domestica). Part II: Dark cells in the pars anterior.
Folia Histochemica Cytochemica 16 (4),315-322.
11. Runceanu L., Cotea C., 2001 Reproducie, obstetric i ginecologie veterinar.
Ed. Ion Ionescu de la Brad, Univ. de tiine Agricole i Medicin Veterinar,
Iai.
12. Vila-Porcile, E., 1972 - Le reseau des cellules folliculo-stellaires et les follicules de
ladenohypophyse du rat (pars distalis). Z. Zellforsch. 129, 328-369.

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NUCLEII HIPOTALAMICI LA VAC


THE NUCLEI OF THE HYPOTHALAMUS COW

C. COTEA C., O.Z. OPREAN,


P.C. BOITEANU, Carmen SOLCAN,
U.S.A.M.V. Iai
I. COTEA
U.M.F. Iai

As a research material we have used the hypothalamico-hypophyseal


complexs of 21 cows in the phases of their sexual cycle. From the 21
samples, we have obtained only ten hypothalamico-hypophyseal complexs;
at the 11 females the hypothalamus has been obtained separately from the
hypophysis, because of the difficulty in getting the complex of the whole.
The samples have been fixed in Orth, Carnoy and Helly, led up to paraffin
and serially sectioned at 5 m. We have finally obtained 550 serial sections
stained through the methods: Novelli, PAS, Fontana, Steedman-Mowry,
Bielschowsky, Bodian, Holmes and Nissl. In the cow hypothalamus we have
found the following groups of neurons: anterior (nuc. supraopticus NSO;
nuc. paraventricularis NPV), middle (nuc. infundibularis), lateral (nuc.
hypothalamicus dorsomedialis; nuc. hypotalamicus ventromedialis) and
posterior (nuc. periventricularis caudalis; nuc. premamilaris; nuc. corporis
mamilaris). The name of the groups has been given according to their site
compared to the hypothalamo-hypophyseal tract. In the walls of the third
hypothalamus ventricle in the cow are localized the neurons which
elaborate the releasing hormones (RH) for the gonadotrophic hormones.
These neurons structure de infundibularis nucleus of the cow hypothalamus.
The neurons are multipolars with an average dimension of 10-12 m, which
a vesiculous nucleus 5 m large and nucleolated. In the neuroplasm of the
axon hillock we have found the numerous PAS positive granules. These
granules are positive in Mikami stain, get stained in blue, which
demonstrates that they are polipeptidic nature. FSH-RH and LH-RH are
polipeptids and stimulate the secretions of the gonadotrophic hormones in
the -FSH and the -LH cells from the cow adenohypophysis.

Key words: cow, hypothalamus, nuclei, releasing hormones (RH)

Hipotalamusul a fost studiat la scroaf i la oaie (1, 5, 7, 8, 9), cu


evidenierea neuronilor secretori de polipeptide FSH-RH i LH-RH. n vederea
descrierii neuronilor din nucleii hipotalamici implicai n reproducie la vaci, s-a
ntreprins prezenta lucrare.

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MATERIAL I METOD

Ca material de cercetat au servit hipotalamusurile i hipofizele de la 21 vaci


n fazele ciclului sexual, recoltate imediat dup sacrificarea animalelor.
Fragmentele au fost fixate n Orth, Carnoy i Helly, incluse n parafin i
secionate la 5 m. Numai la 11 femele s-a obinut complexul hipotalamo-
hipofizar, la restul femelelor hipotalamusul a fost obinut separat de hipofiz, dat
fiind dificultatea recoltrii complexului n totalitatea sa. Din cele 11 piese obinute
s-au efectuat pentru fiecare caz cte 240 seciuni seriate, pentru a urmri neuronii
hipotalamici n funcie de axul hipotalamo-hipofizar i n raport cu ventriculul III i
infundibulul acestuia. Coloraiile folosite au fost: Novelli, PAS, Fontana,
Steedman-Mowry, Bielschowsky, Bodian, Holmes i Nissl.

REZULTATE I DISCUII

n hipotalamus la vac se delimiteaz mai multe regiuni i arii de neuroni.


Importana major n sistemul funcional endocrin o au aceste regiuni cunoscute
sub numele de nuclei hipotalamici (1, 5, 7, 8) care sunt structurai din neuroni
secretori.
n hipotalamus s-au evideniat urmtoarele grupuri de neuroni: anterior,
mijlociu i posterior. Denumirea grupurilor de neuroni a fost dat n raport cu
poziia fiecruia fa de fasciculul hipotalamo-hipofizar, fa de ventriculul III i
infundibulul acestuia (Plana I).
n grupul anterior n raport cu localizarea cmpurilor de neuroni se
evideniaz nucleul supraoptic (NSO) i nucleul paraventricular (NPV) (fig. 1,
2, 3, 4, 5, 6). NSO se remarc deasupra chiasmei optice, n timp ce NPV este
situat anterior ventriculului al III-lea. Neuronii din NSO i NPV sunt de talie mare
(24-31 m), multipolari, cu nucleul veziculos, localizat excentric i nucleolat.
Axonii acestor neuroni, n seciunile impregnate argentic, pot fi urmrii n
cmpul microscopic pe distane de sute de micrometri. n lungul acestor axoni se
remarc granule n iraguri, de diferite dimensiuni i PAS-pozitive. Aceleai
granule se evideniaz i n corpii Herring ai neurohipofizei. n substana
interstiial a lobului posterior se remarc de-alungul fibrelor nervoase formaiuni
ovalare rotunde sau cu contur neregulat, de dimensiuni variabile numite corpii
Herring. Acetia sunt considerai ca fiind forma sub care se stocheaz secreia
neuronilor hipotalamici n lobul posterior, deoarece dup secionarea fasciculului
hipotalamo-hipofizar (9) (totalitatea axonilor din nucleii supraoptici i
paraventriculari ai hipotalamusului), substana Herring se acumuleaz la
extremitatea proximal a neuronului secionat. Fibrele nervoase sunt n majoritate
mielinice i reprezint axonii neuronilor secretori din nucleii hipotalamusului:
nucleul supraoptic (NSO) i paraventricular (NPV). Aceste fibre sunt dispuse
paralel n tija hipofizar, au o dispoziie neregulat n lobul posterior i se termin
prin dilataii n form de mciuc n jurul capilarelor sanguine i a pituicitelor.

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Plana I. Sistemul hipotalamo-hipofizar i epitalamo-epifizar la vac:


- Grupul anterior (1. nuc. supraopticus; 2. nuc. paraventricularis);
- Grupul lateral (3. nuc. hypotalamicus dorsomedialis;
4. nuc. hypotalamicus ventromedialis);
- Grupul mijlociu (5. nuc. infundibularis);
- Grupul posterior (6. nuc. periventricularis caudalis; 7. nuc. premamilaris;
8. nuc. corporis mamilaris)
E epifiza; MI masa intermediar; CO chiasma optic; CM corpul mamilar
*
3, 4, 5, 6, 7 formeaz aria hipofizotrop a hipotalamusului la vac, cu neuroni
multipolari secretori de RH i IH

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Fig. 1. Complexul hipotalamoneurohipofizar Fig. 2. Infundibulul ventriculului al III-lea la


la vac. Col. Novelli; x 100 vac. Impregnare argentic tehnica
Holmes; x 80

Fig. 3. Nucleul supraoptic (NSO) din Fig. 4. Nucleul supraoptic (NSO) din
hipotalamusul de vac. Impregnare hipotalamusul de vac. Impregnare
argentic tehnica Holmes; x 200 argentic tehnica Holmes; x 400

Fig. 5. Nucleul paraventricular (NPV) din Fig. 6. Nucleul paraventricular (NPV) din
hipotalamusul de vac. Impregnare hipotalamusul de vac. Impregnare
argentic tehnica Holmes; x 200 argentic tehnica Holmes; x 200

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Neuronii din nucleul supraoptic secret hormonul antidiuretic (ADH-


Antidiuretic Hormone) sau vasopresina cu aciune antidiuretic prin resorbia apei
la nivelul tubilor contori distali i colectori, reducnd n acest mod diureza. n
schimb, nucleul paraventricular secret ocitocina (OXT-Ocitocin) care stimuleaz
contraciile uterine n iniierea travaliului din timpul parturiiei i contracia
celulelor mioepiteliale ale canalelor galactofore, determinnd prin aceasta
eliminarea laptelui. Neuronii secretori ai celor doi nuclei din hipotalamus au
citoplasma bogat n corpii Nissl, un nucleu mare nucleolat, dendrite scurte i o
prelungire unic-axonul. Totalitatea axonilor acestor neuroni din cei doi nuclei
constituie fasciculul hipotalamo-neurohipofizar. Neuronii secret cei doi hormoni
ce sunt legai de o protein specific (neurofizin) i sunt transportai n lungul
axonilor i depozitai la nivelul lobului posterior al hipofizei, n corpii Herring.
Neurosecreia mai apare i ca material fin granular aglomerat n jurul capilarelor
i fibrelor nervoase terminale. Acest mod de secreie a hormonilor n lungul
fibrelor nervoase se numete neurocrinie.
Morfologic, aceste fapte au fost dovedite prin punerea n eviden a
granulaiilor de secreie n pericarionul neuronilor secretori, n lungul fibrelor
nervoase (axonii acestor neuroni) care merg de la neuroni, prin tija
neurohipofizar, n lobul posterior al hipofizei i la nivelul terminaiilor nervoase
n jurul capilarelor. Coloraiile speciale au mai evideniat n aceti corpi Herring
proteoglicani i acidul L-ascorbic. Eliberarea hormonilor din corpii Herring este
operat de pituicite care i separ de neurofizin i i cedeaz sngelui.
Grupurile laterale sunt situate n raport cu axul medio-sagital al
hipotalamusului, de o parte i de alta a ventriculului III. n structura unui grup
lateral intr doi nuclei: nucleul hipotalamic dorsomedial (fig. 7, 8) i nucleul
hipotalamic ventromedial. Aceti nuclei se deosebesc numai prin poziia lor, n
timp ce sub aspect structural cuprind neuroni cu morfologie similar. Neuronii au
dimensiunea de 14-15 m, sunt multipolari i conin n neuroplasm granule
PAS-pozitive.

Fig. 7. Nucleul hipotalamic dorso-medial din Fig. 8. Nucleul hipotalamic dorso-medial din
hipotalamusul de vac. Impregnare hipotalamusul de vac. Impregnare
argentic tehnica Holmes; x 200 argentic tehnica Holmes; x 200

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Grupul mijlociu se remarc n jurul originii infundibulului ventriculului


III, de unde i denumirea de nucleu infundibular (fig. 9, 10, 11, 12), care este
structurat din neuroni multipolari, cu dimensiunea de 10-12 m i cu numeroase
granule n citoplasm.

Fig. 9. Nucleul infundibular din Fig. 10. Nucleul infundibular din


hipotalamusul de vac. Impregnare hipotalamusul de vac. Impregnare
argentic tehnica Holmes; x 80 argentic tehnica Holmes; x 200

Fig. 11. Nucleul infundibular din Fig. 12. Nucleul infundibular din
hipotalamusul de vac. Impregnare hipotalamusul de vac. Impregnare
argentic tehnica Holmes; x 200 argentic tehnica Holmes; x 400

Grupul posterior include neuronii localizai posterior ventriculului III.


Unii neuroni cu dimensiunea de 10 m sunt dispui n apropierea epiteliului
ependimar al ventriculului III, structurnd nucleul periventricular caudal, n
timp ce ali neuroni apar anterior corpului mamilar, cu dimensiunea de 12 m,
formnd nucleul premamilar (fig. 13, 14).
n corpul mamilar s-au evideniat neuroni multipolari cu dimensiunea de
16-18 m, ce structureaz nucleul mamilar (fig. 15, 16).

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Sistemele de interrelaie ntre hipotalamus i hipofiz sunt: sistemul


hipotalamo adenohipofizar i sistemul hipotalamo-neurohipofizar.

Fig. 13. Nucleul premamilar din Fig. 14. Nucleul premamilar din
hipotalamusul de vac. Impregnare hipotalamusul de vac. Impregnare
argentic tehnica Holmes; x 200 argentic tehnica Holmes; x 200

Fig. 15. Nucleul mamilar din hipotalamusul Fig. 16. Nucleul mamilar din hipotalamusul
de vac. Impregnare argentic tehnica de vac. Col. Nissl; x 200
Holmes; x 200

Sistemul hipotalamo-neurohipofizar este reprezentat de nucleii din zona


hipotalamic anterioar - nucleii supraoptici i paraventriculari. Axonii formeaz
fasciculul hipotalamo-neurohipofizar. Nucleii supraoptici i paraventriculari
secret vasopresina (ADH) i ocitocina (OXT). Aceti doi hormoni sunt legai
de o protein specific - neurofizina - fiind transportai de-a lungul axonilor n
neurohipofiz.
Sistemul hipotalamo-adenohipofizar este caracterizat prin existena
neuronilor secretori din aria hipofizotrop a hipotalamusului care sintetizeaz
hormoni hipofizotropi de stimulare i eliberare (Releasing-Hormones R.H.) i
inhibitori (Inhibiting Hormones -IH) ai secreiilor adenohipofizare.

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n neuronii nucleilor dorsomedial, ventromedial, infundibular,


periventricular caudal i premamilar s-au evideniat granule PAS-pozitive, ceea ce
pledeaz pentru o secreie de natur polipeptidic n aceti neuroni. Ei
structureaz aria hipofizotrop n hipotalamusul de vac, deoarece s-a demonstrat
c hormonii de eliberare i de inhibare (RH i IH) sunt din punct de vedere chimic
polipeptide. Axonii acestor neuroni se evideniaz n jurul capilarelor sanguine
din pereii infundibulului ventriculului al III-lea, unde i elibereaz secreiile
specifice pentru stimularea sau inhibarea celulelor secretoare din adenohipofiz.

CONCLUZII

1. n hipotalamus la vaci se evideniaz urmtoarele grupuri de neuroni:


anterior, laterali, mijlociu i posterior.
2. Grupul anterior este structurat din nucleul supraoptic (NSO) i nucleul
paraventricular (NPV) cu dimensiunea pericarionului de 24 - 31 m.
3. Grupurile laterale sunt structurate din nucleul hipotalamic dorsomedial i
nucleul hipotalamic ventromedial cu neuroni al cror pericarion este de 1415
m.
4. Grupul mijlociu (nucleul infundibular) este structurat din neuroni
multipolari, cu dimensiunea de 10-12 m, localizai n jurul infundibulului
ventriculului al III-lea i sunt secretori ai FSH-RH i LH-RH.
5. Grupul posterior include neuronii cu dimensiunea de 10 12 m,
localizai posterior ventriculului III i sistematizai n nucleul periventricular
caudal i nucleul premamilar.

BIBLIOGRAFIE

1. Cotea C., Arseni O.V., 1994 Hipotalamusul la suine. Lucr. t. Med. Vet. Iai, vol.
37, p. 25 27.
2. Cotea C., Carmen Solcan, Arseni O.V., 1997 Structuri foliculare n lobul anterior al
hipofizei la femelele suine din rasa Landrace. Lucr. t. Med. Vet. Iai, vol. 40, p. 20
22.
3. Cotea C., Arseni O.V., 1998 Foliculogeneza n ovarul suinelor femele. Lucr. t.
Med. Vet. Iai, vol. 41, pag. 13 23.
4. Cotea C., Arseni O.V., Carmen Solcan, 1998 Glanda tecal ovarian la suinele
femele. Lucr. t. Med. Vet. Iai, vol. 41, pag. 6 12.
5. Cotea C., Arseni O.V., 1999 Hipofiza la suinele femele. Lucr. t. Med. Vet. Iai,
vol. 1 (42), p. 24 31.
6. Cotea C., 2001 - Biologie celular, Embriologie general, Histologie general, Ed.
Tehnopress, Iai.
7. Cotea C., Runceanu L., Carmen Solcan, Cotea I., 2002 Neurosecreia nucleilor
hipotalamici la oaie. Lucr. t. Med. Vet. Iai, vol. 45 (4), p. 16 22.
8. Cotea C., 2003 Histologie special. Ed. Tehnopress, Iai.
9. Runceanu L., Cotea C., 2001 Reproducie, obstetric i ginecologie veterinar.
Ed. Ion Ionescu de la Brad, Iai.

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OBSERVAII PRIVIND METAPLAZIA CARTILAGINOAS


I OSOAS A PIELII LA VACI CU CETOZ
OBSERVATIONS CONCERNING CARTILAGINOUS AND OSSEOUS METAPLASIA
OF THE SKIN IN DAIRY COWS WITH KETOSIS

Carmen SOLCAN, O.Z. OPREAN,


C.V. COTEA, Gh. SOLCAN,
U.S.A.M.V. Iai

In 15 dairy cows slaughtered because of ketosis the histological


lesions of the skin from different regions (periorbital, sternum, base of the
tail and ventral abdominal) were studied. On the skin from sternum region,
chondral and osseous metaplasia was observed in 3 of cases (20%). The
new formed osseous tissue had a structure like a spongious osseous tissue.
Degenerative and metaplasic lesions of the skin were related with
degenerative lesions of the liver and kidney

Key words: dairy cows, ketosis, skin, osseous metaplasia

Metaplazia osoas a pielii se caracterizeaz prin transformarea esutului


conjunctiv dermic n esut osos. Fibrele de colagen se transform n esut osteoid
iar fibroblastele n osteoblaste (5). La vaci este rareori semnalat n literatura de
specialitate.
Cetoza vacilor pentru lapte este consecina unei perturbri progresive a
metabolismului energetic, constnd n insuficiena cronic a aportului de glucide
uor digestibile, tradus prin instalarea treptat a unor leziuni ale organelor interne
i ale glandelor endocrine implicate n metabolismul energetic, constnd n
steatoz hepatic i renal, hiperfuncie hipofizar i corticosuprarenal i
hipofuncie tiroidian. S-a urmrit depistarea leziunilor cutanate asociate cetozei.

MATERIAL I METODE

S-au efectuat investigaii morfologice ale pielii la 15 vaci sacrificate de


necesitate datorit cetozei. Recoltarea fragmentelor de piele s-a realizat din
urmtoarele regiuni corporale: periorbital, stern, baza cozii, regiunea
abdominal. Recoltrile s-au efectuat n luna iunie. Piesele au fost prefixate n
formaldehid 10% soluie apoas, timp de 12 ore, apoi n fixator Bouin timp de 48
de ore, apoi incluse n parafin i secionate la 5m. Au fost efectuate coloraii
HEA, Acid Periodic Fuxin Schiff, Giemsa pe esuturi i Pappenheim. S-au
efectuat investigaii morfometrice i histochimice urmrindu-se grosimea
epidermei, infiltraiile limfoide, particulariti morfologice i modificrile histologice
ale pielii n regiunile cu leziuni.

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REZULTATE I DISCUII

Pielea din regiunea sternal i de la baza cozii prezenta leziuni


macroscopice locale de hiper- i paracheratoz i depilaii difuze.
Pielea din regiunea sternului i a bazei cozii se caracterizeaz printr-o
hipercheratoz pronunat. La nivelul epidermei se contat prezena spiculilor
hiperkeratozici, iar n derm absena glandelor sebacee, sudoripare i a foliculilor
piloi (fig. 1). La 3 cazuri (20%) s-au ntlnit diferite stadii de evoluie a
metaplaziei osoase. ntr-un prim stadiu dermul este foarte dens, bogat celule,
respectiv fibroblaste care sunt implicate n sinteza fibrelor de colagen i a
substanei fundamentale (fig. 2). Fibroblastele se metaplaziaz n condroblaste
care vor structura esutul cartilaginos elastic (fig. 3). Ulterior are loc depunerea
srurilor de calciu i magneziu care duc la calcificarea cartilajului (6, 7, 8).
esutul osos format este asemntor celui spongios (fig. 4), dar prezint o cretere
a densitii osului datorit fibrelor conjunctive (osteoscleroz), corelat cu
disfuncia hepatic. n areolele esutului osos sunt prezente elemente figurate
dintre care predomin hematiile i plachetele sanguine situate pe un citoreticul.

Fig. 1. Piele din regiunea sternului. Spiculi hiperkeratozici la nivelul epidermei. Derm
bogat n fibroblaste. Col PAS; x 200; x 400

Modificrile pielii din aceaste zone (stern i baza cozii) se pot corela dup
unii autori (6, 8) cu disfunciile hepatice i renale, la cazurile noastre acestea fiind
determinate de cetoz, caracterizndu-se prin hepatoz i nefroz gras
pronunat.

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Fig 2. Piele din regiunea sternului. Activitatea intens a fibroblastelor de sinteza a


fibrelor de colagen i a substanei fundamentale. Col. Giemsa; x 100; x 200.

Fig.3. Piele din regiunea sternului. esut cartilaginos elastic n derm. Col. PAS; x400.

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Fig. 4. Piele din regiunea sternului. esut osos cu travee dense, fibroase (osteoscleroz).
Col PAS; 100; x200x 400.

Epiderma n regiunea periorbital i la nivelul pleoapei superioare i


inferioare are un diametru de 120-134 m, iar dermul mpreun cu glandele
Meibomius un diametru de 2500-2800 m. n lamina propria a acestor canale i n
dermul pleoapelor apar infiltraii limfoide difuze. Alturi de limfocite se pot
evidenia i plasmocite.
Pielea din regiunea pliului iei prezint epiderma cu un diametru de 120 m,
iar mpreun cu dermul are o grosime de 1020- 1570 m. Glandele sebacee,
sudoripare i foliculii piloi au dispoziie oblic, n unghi ascuit de 30-40. n
stratul papilar al dermului se observ aglomerri limfoide sub forma unor insule
formate din limfocite, plasmocite, eozinofile, rare mastocite i plasmocite.

CONCLUZII

1. Se semnaleaz leziuni histologice de metaplazie cartilaginoas i osoas


a pielii din regiunea sternului i a bazei cozii la 3 din 15 vaci cu cetoz (20%),
leziuni care sunt mai rar menionate la bovine.
2. Metaplazia cartilaginoas i osoas poate fi pus pe seama disfunciilor
hepatice i renale existente la bovinele cu cetoz.
3.esutul osos format este asemntor celui spongios, dar prezint o
cretere a densitii osului datorit fibrelor conjunctive (osteoscleroz).

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BIBLIOGRAFIE

1. Cotea C., 2003 - Histologie Special. Ed. Tehnopress, Iai.


2. Gengoux Vincke P., 1990 - Immunite de la peau in Pastoret P.P., Govaerts A.,
Bazin H., Immunologie animale - Flammarion, Paris, 239-241
3. Gross T.L., Ihrke P.J., Walder E.J., 1992 Veterinary Dermatopathology. A
macroscopic and microscopic evaluation of canine and feline skin disorders.
Mosby Yearbook, St. Louis
4. Mitrea I.L., Solcan Gh., 2006 - Dermatopathologia animalelor de ferm, Ed
Medical Veterinar, Bucureti
5. Paul I., 1989 Morfologia proceselor patologice fundamentale, Ed. Soc. Med. Vet.
din Romnia, Bucureti
6. Ruiz Genao D.P., Pascual Lopez M.P., Fraga S., Aragues M., Garcia Diez A.,
2005 - Osseous metaplasia in the setting of nefrogenic fibrosing dermathopathy.
Pub. Med., Febr., 32(2); 172-5.
7. Scott D.W., 1988 - Large animal dermatology, WB Saunders Co, Philadelphia
8. Yager J.A., Scott D.W., 1993 - The Skin and Appendages n Patology of Domestic
th
Animals.4 ed., vol.I., 531-738, ed. by Jubb K.V.F., Kennedy P.C., Palmer N.,
Academic Press Inc, New York.

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HISTOCHIMIA SINUSULUI INTERDIGITAL LA OAIE


HISTOCHEMICAL STUDY OF THE INTERDIGITAL SINUS (BIFLEXUS GLAND)
IN SHEEP

A. POPESCU, C. COTEA, Carmen SOLCAN


U.S.A.M.V. Iai

For this paper, 72 interdigital sinuses served as study, they have


been collected immediately after the animal's sacrification immediately.
These glands were sectioned transversally or medio-sagittally, so it
was possibe to obtain from each gland 7 fragments: A, B, C, D, E, F, G.
For the histochemical study there were used 18 glands that were
sectioned medio-sagittally, and 54 were sectioned conforming to the
areas,obtaining 378 samples. These were enclosed in paraffin and cut into
five-micrometer thick pieces.
The stains were: PAS, PAS- Alcian blue, Novelli, Papanicolau.
1584 slides which have been interpreted and photographed at light
microscope.
We have identified numerous hair follicles, and nearby them
sebaceous glands. Sinuous and tubular glands which form interdigital gland
are grouped in glandular lobules separated by thin conjunctival septa.
The secretion of the gland is abundant in proteoglicans and cells
present secretor cycle, relieving the accumulation stage of PAS-positive
secretion especially.

Key words: interdigital sinus, sheep, tubular gland.

Din bibliografia consultat rezult o serie de date histologice i


histochimice contradictorii privind glanda biflex la ovine, fapt ce a determinat
ntreprinderea prezentei lucrri. Acest studiu era impetuos, innd cont de faptul
c n literatura de specialitate nu se gsesc date referitoare la glanda biflex la
ovinele din rasa Karakul brumariu. S-a efectuat n premiera o zonare a glandei,
efectundu-se numeroase seciuni medio-sagitale i transversale prin urmtoarele
7 zone: A, B, C, D, E, F, G (Fig. 1).

MATERIAL I METODE

Glandele au fost disecate att la membrele anterioare ct i la cele


posterioare imediat dup sacrificare i apoi s-a efectuat zonarea, rezultnd 7
zone (A, B, C, D, E, F, G) (Fig. 1).
Coloratiile folosite au fost: PAS, PAS- albastru alcian, Novelli, Papanicolau.
S-au obtinut 1584 preparate permanente ce au fost citite si fotografiate la
microscop
Au fost obinute 30 de microfotografii color.

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Fig. 1. Glanda biflex (sinusul interdigital) la oaie, schi cu seciunile transversale din care
s-au recoltat fragmente pentru prelucrarea histologic A, B, C, D, E, F, G:
Po - poriunea orizontal (A); Pd - poriunea descendent (B, C);
Pa - poriunea ascendent (E, F, G);
A - poriunea orizontal (treimea superioar); B - treimea mijlocie;
C - treimea inferioar; D - jonciunea dintre Pd i Pa;
E - treimea inferioar; F - treimea mijlocie; G - treimea superioar (fundul de sac).

Glanda biflexa - zona A


Pe seciuni transversale se remarc lumenul uor festonat, cu o secreie
abundent. Epiteliul luminal este pavimentos stratificat cheratinizat, n care la
suprafa apar lame desprinse de cheratin.
Epiteliul se sprijin pe un derm n care se evideniaz foliculi piloi, n
vecintatea crora apar glande sebacee (Plana I, fig. 3, 4).
Glandele sudoripare sunt foarte dezvoltate. Acestea sunt mai rare, satelite
foliculilor piloi, formate din uniti secretoare de diferite forme i dimensiuni.

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Glandele sebacee apar cu o structur caracteristic de sac. De obicei apar


grupate n apropierea foliculilor piloi. Ele se remarc n apropierea epidermului.
n jurul orificiului de deschidere a glandei biflexe s-au remarcat foliculi piloi de
dimensiuni mari, a cror tij formeaz smocuri de peri (Plana I, fig. 6). n
vecintatea foliculilor piloi se remarc glande sebacee, care prin secreia lor
asigur lubrefierea tijelor din smocul de peri (Plana I, fig. 5).
n profunzime apar glande sudoripare, asemntoare cu cele ale pielii ca
structur. Lobulii structurai din glandele sudoripare se remarc n profunzimea
dermului, cu celule secretoare n diferite faze ale elaborrii secreiei. Unele glande
sunt pline cu secreie PAS-pozitiv.

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Glanda biflex zona B


n lumenul glandei biflexe apare o secreie abundent PAS-pozitiv.
Epiteliul luminal este pavimentos stratificat cheratinizat, la care stratul
cornos este gros (Plana II, fig. 8). n structura dermului se remarc numeroase
fibre de colagen i foliculi piloi, n jurul crora apar glande sebacee dezvoltate
(fig. 1). n unele zone, adiacente foliculilor piloi dezvoltai, se evideniaz foliculi
piloi solitari cu diametrul mare (Plana II, fig. 9, 11). Unele glande sebacee s-au
observat n apropierea epiteliului luminal, pavimentos stratificat cheratinizat
(Plana II, fig. 10). Foliculii piloi foarte dezvoltai asigur tijele groase din
smocul de peri din jurul orificiului.

Glandele sebacee sunt de tip holocrin, cu un strat generator evident. n


plana II, figura 9 se evideniaz un folicul pilos cu diametru mare. Foliculii piloi
n unele seciuni apar numeroi, i n teaca lor extern se remarc fibre musculare
netede, ce structureaz muchiul erector al firului.

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Glandele sudoripare sunt situate mai n profunzime. Se grupeaz n mici


lobuli. n cadrul lobulului, unitatea secretoare poate fi surprins n diferite faze de
secreie. Astfel se remarc glande sudoripare n faza de acumulare a secreiei n
epiteliul glandular care devine nalt i cu citoplasma apical de aspect vacuolar.
Nucleii celulelor sunt veziculoi i nucleolai. n jurul unitilor secretoare se
remarc fibre musculare netede.

Glanda biflex zona C


Secreia apare abundent n lumen, PAS-pozitiv i omogen, uneori
aceast secreie ader intim de epiteliul pavimentos stratificat cheratinizat.
Epiteliul ce delimiteaz lumenul glandei n aceast zon este pavimentos
stratificat cheratinizat, la care stratul cornos este evident, pe unele zone se observ
desprinderea lamelelor de cheratin. n derm se remarc foliculi piloi cu
diametru mare.

Glandele sudoripare sunt grupate n lobuli separai printr-un esut


conjunctiv delicat (Plana III, fig. 13, 14, 15, 16). Uneori, adenomerul glandei
sudoripare prezint pinteni, pentru a mri suprafaa de secreie a glandei.

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Formaiunile secretoare au aspect diferit pe seciunea transversal i conin n


citoplasma celulelor epiteliale glandulare numeroase granule de secreie. Aceast
secreie PAS-pozitiv (Plana III, fig. 13, 14) umple pn la refuz formaiunile
glandulare. Unele glande sunt surprinse n faza de elaborare a secreiei,
caracterizat printr-o exocitoz masiv a produsului de secreie. esutul
conjunctiv delicat dintre formaiunile secretoare este intens vascularizat, ceea ce
sugereaz o aprovizionare cu substane nutritive la un nivel ridicat a glandei.
Vasele sanguine sunt reprezentate prin arteriole cu media format din dou-trei
rnduri de leiocite, i venule, n peretele crora se evideniaz leiocite izolate. n
vecintatea laminei bazale a formaiunilor secretoare se remarc reele de capilare
sanguine, care prin transcitoz asigur metabolismul intens al celulelor secretoare.

Glanda biflex zona D

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Lumenul este delimitat de epiteliul pavimentos stratificat cheratinizat, la


care stratul cornos se deprinde n lambouri. n lumenul glandei apare secreia
abundent, omogen i PAS-pozitiv, desprins de epiteliul pavimentos stratificat
cheratinizat ce delimiteaz zona intern. n cadrul unor seciuni histologice a fost
observat secreia de aspect vacuolar. n unele zone stratul cornos este foarte
dezvoltat, iar stratul granular al lui Unna conine n citoplasma celulelor granule
ncarcate cu cheratohialin, iar stratul lucidum prezint granule de eleidin.
Foliculii piloi din aceast zon sunt mai rari i cu diametru mare. n vecintatea
lor se remarc glande sudoripare grupate n lobuli (Plana IV, fig. 18, 19).
Unitile secretoare ale glandelor sudoripare pot fi surprinse n diverse faze de
secreie. n funcie de aceste faze, epiteliul secretor prezint morfologie
caracteristic. n lumenul adenomerului se remarc o secreie crescut PAS-
pozitiv (Plana IV, fig. 20, 21), ce rezult n urma exocitozei materialului
secretor ce se acumuleazla polul apical al celulelor epiteliale glandulare.

Glanda biflex zonele E si F


Secreia apare n exces n aceast zon, evideniat prin faptul c unitile
secretoare sunt pline cu produsul de secreie PAS-pozitiv (Plana V, fig. 23, 24).
Uneori, secreia apare de aspect vacuolar n lumenul formaiunilor secretoare.
Unitile secretoare prezint numeroi pinteni, epiteliul secretor este nalt i
manifest semne intense de exocitoz. Formaiunile secretoare sunt dezvoltate,
realiznd un strat glandular compact. Epiteliul glandular este cubic sau nalt, n
funcie de faza n care sunt surprinse formaiunile glandulare. Dup eliberarea
secreiei, epiteliul glandular este cubic i se pregtete pentru refacerea
organitelor citoplasmatice n vederea relurii unui nou ciclu secretor
Lobulii glandulari sunt aezai pe dou-trei rnduri structurnd un strat
glandular compact. Pe seciune, unitile secretoare prezint lumenul ncarcat cu
secreie omogen i PAS-pozitiv (Plana V, fig. 25, 26). Unii lobuli sunt
surprini n faza n care secreia este eliminat din cadrul formaiunilor
glandulare. Au fost evideniai lobuli n care procesul secretor se desfoar la un
nivel ridicat n unitile secretoare, a cror epiteliu ncepe s devin prismatic
simplu. n ali lobuli, epiteliul glandular este aplatizat, datorit acumulrii n exces
a secreiei PAS-pozitiv.
n lumenul glandei, secreia este vscoas, PAS-pozitiv i ader intim de
epiteliul luminal pavimentos stratificat cheratinizat, la care procesul de
cheratinizare este mai puin intens dect n celelalte zone, demonstrat prin
acumularea unor granule mai rare de cheratohialin i eleidin.

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Glanda biflex zona G


Formaiunile secretoare sunt dezvoltate, realiznd un strat glandular
compact (Plana VI, fig. 28, 29, 30, 31). Epiteliul glandular este prismatic, n
unele zone cubice, cu o secreie abundent PAS-pozitiv n lumen (Plana VI, fig.
29).
Unele formaiuni glandulare prezint pinteni epiteliali pentru mrirea
suprafeei de secreie. Celulele epiteliale prezint n citoplasma polului apical
cantiti crescute de granule de secreie care sunt eliminate prin exocitoz (Plana
VI, fig. 29, 30).

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REZULTATE I DISCUII

Din studierea sistematic a celor 7 zone ale glandei biflexe la oaia Karakul
brumriu, se evideniaz diferene structurale ntre ele. n primul rnd, pe msura
apropierii de zona G se constat dezvoltarea deosebit a stratului glandular ce
intereseaz glandele sudoripare, ceea ce ne determin s afirmm c glanda
biflex este o gland sudoripar modificat, n sensul c unitile secretoare pe
lng faptul c excret, secret i o cantitate crescut de material PAS-pozitiv.

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Prezena glandelor sudoripare ca formaiuni secretoare de diferite forme n


seciunile histologice, demonstreaz ca morfologie aspectul tubulo-sinuos.
Formaiunile secretoare sunt n numr variabil n cadrul unui lobul, ce este
delimitat de un esut conjunctiv delicat i foarte vascularizat.
Histochimic s-a demonstrat prezena att n lumenul glandelor sudoripare,
ct i n cel al polului apical al celulelor secretoare, a proteoglicanilor, ceea ce
demonstreaz c secreia acestor glande este foarte bogat n glicoproteine, n care
fraciunea proteic este bine reprezentat, aspect menionat i de ali autori (2, 3,
4, 7).
n cadrul aceluiai lobul, unitile secretoare pot fi surprinse n diferite faze
ale secreiei. Prezena unui epiteliu nalt, cu aglomerare de proteoglicani la polul
apical al celulelor secretoare, demonstreaz faza de acumulare a secreiei, stadiul
presecretor. n unele formaiuni secretoare ale glandei biflexe s-a evideniat
secreie PAS-pozitiv ce umple pn la refuz lumenul acestora. Epiteliul
glandular n acest caz este cubic sau de cele mai multe ori aplatizat, indicnd
stadiul postsecretor al glandelor sudoripare. Modalitatea de secreie este apocrin;
dup ce secreia se acumuleaz la polul apical, celulele sufer procesul de
decapitare, asemntoare celulelor din glanda mamar (2, 3, 4, 7). Unele
formaiuni glandulare prezint pinteni sau pliuri pe seciunea transversal cu rolul
de a mri suprafaa de secreie a epiteliului. Printre celulele epiteliale ataate de
lamina bazal s-au remarcat celule mioepiteliale cu rol n elaborarea secreiei, prin
sistemul motil actin-miozin.
Cei mai dezvoltai lobuli glandulari se remarc n zonele F i G, ceea ce
demonstreaz c aceste zone particip cel mai intens la elaborarea secreiei.
Faptul c foliculii piloi au diametrul mai mare n zona de deschidere a
glandei biflexe, demonstreaz grosimea mai mare a tijelor smocului de peri din
aceast regiune.
Glandele sebacee au structur de sac alveolar cu lamina bazal pe care se
sprijin stratul generator, i straturi celulare implicate n procesul de holocrinie. n
aceste celule s-au evideniat cantiti variabile de lipide, sub form de picturi
mari n apropierea zonei centrale de necrobioz. Ele sunt mai frecvente n zona A
i zona B, n apropierea epiteliului cheratinizat. n ceea ce privete epiderma
(epiteliul pavimentos stratificat cheratinizat), ce cptuete lumenul glandei
biflexe, este mai groas, cu stratul cornos mai evident n zona orificiului de
deschidere a glandei. Epiteliul cheratinizat se pstreaz pe toat lungimea glandei,
n care s-au evideniat granule de cheratohialin i eleidin dispuse n irag. n
unele zone ale lumenului glandular se remarc procesul de exfoliere n lambouri a
lamelelor de cheratin. Secreia este omogen, intens PAS-pozitiv i ader intim
de epiteliul luminal pavimentos cheratinizat. n unele zone, materialul secretor
apare intens vacuolar sau de aspect retiform, n sensul c secreia ia aspect
filamentar la punctul de contact cu epiteliul luminal.

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CONCLUZII

1. Glanda biflex la ovinele din rasa Karakul negru este o gland sudoripar
modificat, n care formaiunile secretoare se organizeaz n lobuli.
2. Lobulii sunt delimitai de un esut conjunctiv delicat, foarte vascularizat
i n cadrul lor, unitile secretoare sudoripare pot fi surprinse n diferite faze ale
secreiei.
3. Cea mai dezvoltat zon glandular este fundul de sac al glandei biflexe
(zona E), unde lobulii glandulari sunt mai numeroi i mai voluminoi,
manifestnd o activitate secretorie intens.
4. Procesul de elaborare a secreiei acestor glande este de tip apocrin, cu
morfologia caracteristic ce decurge din aceasta.
5. Evidenierea proteoglicanilor n secreia luminal, ct i la polul apical al
celulelor glandulare, demonstreaz c secreia glandei conine cantiti crescute de
proteine n cadrul compuilor glicoprotidici.
6. Lumenul glandei biflexe este delimitat de un epiteliu pavimentos
stratificat cheratinizat cu stratul cornos mai bine reprezentat n primele 3 zone i
care se menine de tip cornos pe toat suprafaa luminal a glandei.
7. Glandele sebacee sunt prezente n vecintatea foliculilor piloi, mai ales
n zonele A i B ale glandei biflexe.
8. Foliculii piloi sunt mai dezvoltai n zona A, unde formeaza tijele groase
ale smocului de peri n zona orificiului.
9. Prezena a numeroase glande sebacee n zonele A i B, demonstreaz o
secreie bogat n lipide, ce asigur lubrefierea smocului de peri din zona
orificiului de deschidere a glandei biflexe.

BIBLIOGRAFIE

1. Botrel S., Cotea C., Gaboreanu M., 1982 - Histologie i embriologie medical
veterinar, Ed. Didactic i Pedagogic, Bucureti.
2. Cornil L., Manolescu N., 1995 - Structura i ultrastructura organelor la animalele
domestice, Editura Ceres, Bucureti.
3. Cotea C., Gall C., 1985 - Aspecte histologice i histochimice ale glandei biflexe la
ovine. Seminarul Tehnologia reproduciei i patologia animalelor de ferm, vol.
X, p. 331-336, Institutul Agronomic Cluj-Napoca.
4. Cotea C., 2003 - Histologie special. Ed. Tehnopress, Iai.
5. Coofan V., 1975 - Anatomie topografic a animalelor domestice. Lito., vol. II,
Institutul Agronomic Iai.
6. Coofan V., Palicica R., Valentina Hricu, Enciu V., 1999 Anatomia animalelor
domestice, vol. I. Aparatul de susinere i micare. Ed. Orizonturi universitare,
Timioara.
7. Grigorescu E., Moisescu M., Olarian Elena, Vcaru C., 1957 Contribuii la studiul
sinusului interdigital la oaie. Lucrrile Sesiunii tiinifice, Institutul Agronomic
Timioara, p. 273-281, 1957.

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8. Janicki Z., Hrarste A., Slavica A., Konjevi D., Marinovi Z., Stubian D., 2003 -
Morphohistological characteristics of the interdigital gland in the roebuck
(Capreolus capreolus L.). Vet. Arhiv 73, 27 37.
9. Knig E.H., Liebich G.H., 1999 Anatomie der Haussugetiere. Lehrbuch und
Farbatlas fr Studium und Praxis. Band I, Bewegungsapparat Schauttauer
Stuttgart, New York.
10. Knig E.H., Liebich G.H., 1999 Anatomie der Haussugetiere. Lehrbuch und
Farbatlas fr Studium und Praxis. Band II, Organe, Kreislauf-und Nervensystem,
Schauttauer Stuttgart, New York.
11. Liebich H.G., Bck P., Budras K.D., Maierl J., Reese S., 1999 Funktionelle
Histologie der Haussugetiere. Lehrbuch und Farbatlas fr Studium und Praxis.
Schattauer Stuttgard, New York.
12. Mihai M., 1984 Cercetri histologice, citologice, ultrastructurale i morfochimice
asupra glandelorsebacee i sudoripare n decursul ciclului estral i al gestaiei la
ovine. Tez de doctorat, Institutul Agronomic Iai.
13. incai Mariana, 2000 Histologie veterinar, vol. II, Organe i embriologie. Ed.
Mirton, Timioara.
14. Verde N., Pop T., Manolescu N., 1982 Contribuii la ontogeneza i structura
sinusului interdigital la oaie. Seminarul tiinific "Metode noi de sporire a
produciei la animale" Iai, p. 66-67, 1982.

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MORFOLOGIA SINUSULUI INTERDIGITAL


(GLANDEI BIFLEXE) LA OAIE
MORPHOLOGY OF THE INTERDIGITAL SINUS (BIFLEXUS GLAND) IN SHEEP

A. POPESCU, C. COTEA, Carmen SOLCAN


U.S.A.M.V. Iai

The extremity of the thoracic and pelvic limbs from 18 sheep black
Karakul breed beetwen 3 and 8 years old were sectioned for observation of
the interdigital sinus.
This apears like a pipe that opens through an orifice of 2-4,5 mm
diameter,which is situated in the interdigital space of the dorsal side of
acropodyal area.
The gland was systematised in 4 areas: horizontal, descendent, the
jonction of two areas, ascendent.
The total length of the gland for frontal limbs is beetwen 36,5 mm,
but it may vary beetwen 31-44 mm.
For the posterior limbs, the length can amount to 25- 36 mm, with an
average of 33 mm.
In the present work presented are shown the dimensions of the
studied areas that may vary depending on the limb and age.

Key words: interdigital sinus, sheep, limb.

Datele din literatura de specialitate privitor la marimea glandei biflexe la


oaie, la functia sa si ontogeneza prezinta date contradictorii. (2, 3, 4, 5, 7, 10, 12,
13, 14)
Dimensiunile acestei glande variaza dupa diferiti cercetatori, dar nu se
refera la aceiasi rasa in functie de varsta. (7)
Avand in vedere aceste aspecte contradictorii din bibliografia consultata, s-
a intreprins prezenta lucrare.

MATERIAL SI METODA

Ca material de studiat au servit glandele interdigitale atat de la membrele


anterioare cat si de la cele posterioare de la 18 ovine din rasa Karakul brumariu in
varsta de 3-8 ani sacrificate de necesitate, cate trei pentru fiecare categorie de
varsta.
Regiunile acropodiale de la toate membrele au fost disecate pentru
evidentierea glandei biflexe in vederea fotografierii (fig. 1) si efectuarii unor schite
clare privind topografia acestei glande (fig. 2), deoarece in literatura de
specialitate nu se prezinta clar morfologia acestei glande.

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Fig. 1 Glanda biflexa la oaia Karakul brumariu in varsta de 5 ani:


a - orificiul de deschidere, b - sinusul interdigital

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Fig. 2. Glanda biflex (sinusul interdigital) la oaie:


a - orificiul de deschidere; b - sinus interdigital; c - falanga proximal;
d - ligament interdigital; e - extremitatea distal a metacarpului; Po - poriunea orizontal;
Pd - poriunea descendent; Pa - poriunea ascendent.

REZULTATE SI DISCUTII

Greutatea in grame si dimensiunea glandei biflexe in milimetrii si pe zone


la ovinele din rasa Karakul brumariu in varsta de 3-8 ani sunt redate in Anexele:
1, 2, 3, 4.
In privinta greutatii glandei de la membrul anterior drept,aceasta variaza de
la 8-9 g la varsta de 3 ani la 11-12 g la varsta de 8 ani.
Lungimea totala a glandei variaza de la 31-32 mm la varsta de 3 ani la 43-
44mm la varsta de 7-8 ani.
In cadrul principalelor segmente ( portiunea orizontala -Po ; descendenta-
Pd ; ascendenta -Pa ) se constata ca portiunea orizontala are aceiasi lungime de
2mm si latime de 2mm la toate cele 18 cazuri.
In privinta portiunii descendente se constata ca aceasta variaza ca lungime
de 16-17 mm la varsta de 3 ani la 23-25 mm la varsta de 7-8 ani.Latimea sa este
constanta de 3mm , cu atingerea unei dimensiuni de 4mm la varsta de 7-8 ani.

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La portiunea ascendenta care este cea mai dezvoltata sub aspectul


volumului se constata ca lugimea este de 13-14 mm la varsta de 3 ani ,atingand
17-18 mm la 7-8 ani.Latimea glandei este de 5-7 mm la varsta de 3 ani ,in schimb
la varsta de 7-8 ani este de 7-8 mm.
Greutatea glandei de la membrul anterior stang variaza intre 8-9 g la varsta
de 3 ani la 10-13g la varsta de 7-8 ani.
Lungimea sa totala variaza intre 30-32 mm la varsta de 3 ani la 42-44 mm
la varsta de 8 ani.
Portiunea orizontala are dimensiuni constante lungimea si latimea de 2 mm
indiferent de varsta.
Lungimea portiunii descendente variaza intre 15-16 mm lungime si 3 mm
latime la varsta de 3 ani la 22-24 mm lungime si 3-4mm latime la varsta de 7-8
ani.
Portiunea ascendenta prezinta lungimea de 12-16 mm si latimea de de 5-6
mm la varsta de 3 ani si de 16-19 mm lungime si 5-7 mm latime la varsta de 7-8
ani.
Membrul posterior drept prezinta variatii ale greutatii glandei cuprinse intre
6-7 g pentru varsta de 3 ani si 10-12 g pentru varsta de 7-8 ani.
Lungimea sa totala variaza intre 25-27 mm la varsta de 3 ani pana la 32-36
mm la varsta de 8 ani.
Lungimea si latimea portiunii orizontale ramane aceiasi de 2 mm indiferent
de varsta .
Lungimea portiunii descendente ests cuprinsa intre 13-16 mm la 3-4 ani si
18-21 mm la varsta de 7-8 ani.Latimea sa este cuprinsa intre 2,5-3 mm cu
dimensiunea cea mai mica la doua cazuri la varsta de 5 ani.
Portiunea ascendenta are lungimea de 10-11 mm la varsta de 3-4 ani si de
11-15 mm la varsta de 7-8 ani. Latimea sa este de 4-6 mm indiferent de varsta.
Greutatea glandei la membrul posterior stang variaza intre 6-8 g la varsta
de 3-4 ani la 10-12 g la varsta de 8 ani.
Lungimea sa totala este de 25-28 mm la 3-4 ani si de 32 -36 mm la 7-8 ani.
Portiunea orizontala prezinta lungimea si latimea de 2 mm ce ramane
constanta.
Portiunea descendenta are lungimea de 13-16 mm la 3-4 ani si de 18-20
mm la 7-8 ani. Latimea variaza intre 2,5-3 mm.
Portiunea ascendenta are lungimea de10-11 mm la 3-4 ani si de 12-14 mm
la 7-8 ani. Latimea se incadreaza intre 4- 6 mm la toate varstele.
Din analiza tabelelor 1, 2, 3, 4 se constata ca dimensiunile glandei de la
membrele anterioare respectiv lungimea totala 31-32 mm la varsta de 3-4 ani si
43-44 mm la varsta de 8 ani sunt mai mari decat la membrele posterioare drept si
stang respectiv 25-29 mm la 3-4 ani si de 32-36 mm la varsta de 7-8 ani.
Corespunzator acestor diferente, si greutatea glandei este mai mare la
membrele anterior, variind intre 8-13 g, iar la cele posterioare intre 6-12 g.

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In privinta zonelor luate in studiu: portiunea orizontala , descendent si


ascendenta ultimile doua variaza ca lungime si latime in timp ce portiunea
orizontala este constanta pentru toate varstele.
Aceasta zona se afla in imediata vecinatate a deschiderii glandei si are o
lungime si o latime constanta de 2 mm.
Portiunea descendenta care se intinde de la portiunea orizontala pana la
ligamentul interdigital, avand directie oblica dinaint-inapoi si de sus in jos este
mai mare la membrele anterioare 15-18 mm lungime la 3-4 ani si 22-25 mm la 8
ani fata de dimensiunile de la membrele posterioare 13-16 mm la 3-4 ani si 18-21
mm la 7-8 ani.
Latimea glandei pe toata lungimea portiunii distale este costanta de 2,5-3
mm pentru toate varstele.
Portiunea ascendenta este mai scurta ca cea descendenta. Pozitia este oblica
de jos in sus si dinapoi inainte, portiunea terminala, prin peretele sau infero-
anterior venind partial in raport direct cu fata posterioara a portiunii descendente.
Aceasta are o latime mai mare, de 5-7 mm la 3 ani si de 5-8 mm la 8 ani la
membrele anterioare fata de 4-5 mm la varsta de 3-4 ani si de 4-6 mm la 8 ani la
cele posterioare.
Lungimea variaza intre 12-16 mm la 3-4 ani si 16-19 mm la varsta de 7-9
ani la membrele anterioare fata de 10-11 mm la varsta de 3-4 ani si 11-15 mm la
varsta de 7-8 ani la cele posterioare.

CONCLUZII

1. Glanda interdigitala examinata la 18 ovine din rasa Karakul brumariu in


varsta de 3-8 ani a prezentat variatii in ceea ce priveste greutatea si lungimea
totala a glandei in functie de varsta si de bipedul anterior fata de cel posterior .
2. Greutatea glandei prezinta mici variatii intre membrele anterioare si cele
posterioare totusi ea este mai mare la membrele anterioare.
3. Lungimea totala este mai mare la membrele anterioare decat la cele
posterioare.
4. Glanda interdigitala este sistematizata in 3 zone: portiunea orizontala ;
portiunea descendenta ; portiunea ascendenta.
5. Portiunea orizontala la toate varstele luate in studiu atat la bipedul
anterior cat si la cel posterior prezinta o lungime de 2mm si o latime tot de 2mm
constante pentru toate varstele.
6. Lungimea portiunii descendente este mai mare la membrele anterioare
fata de cele posterioare.
7. Latimea portiunii descendente este mai mare la membrele anterioare fata
de cele posterioare.
8. Lungimea portiunii ascendente este mai mare la membrele anterioare mai
ales la membrul stang decat la cele posterioare.

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9. Latimea portiunii ascendente este mai mare la membrele anterioare in


special la membrul drept fata de cele posterioare.

BIBLIOGRAFIE

1. Botrel S., Cotea C., Gaboreanu M., 1982 - Histologie i embriologie medical
veterinar, Ed. Didactic i Pedagogic, Bucureti.
2. Cornil L., Manolescu N., 1995 - Structura i ultrastructura organelor la animalele
domestice, Editura Ceres, Bucureti.
3. Cotea C., Gall C., 1985 - Aspecte histologice i histochimice ale glandei biflexe la
ovine. Seminarul Tehnologia reproduciei i patologia animalelor de ferm, vol.
X, p. 331-336, Institutul Agronomic Cluj-Napoca.
4. Cotea C., 2003 - Histologie special. Ed. Tehnopress, Iai.
5. Coofan V., 1975 - Anatomie topografic a animalelor domestice. Lito., vol. II,
Institutul Agronomic Iai.
6. Coofan V., Palicica R., Valentina Hricu, Enciu V., 1999 Anatomia animalelor
domestice, vol. I. Aparatul de susinere i micare. Ed. Orizonturi universitare,
Timioara.
7. Grigorescu E., Moisescu M., Olarian Elena, Vcaru C., 1957 Contribuii la studiul
sinusului interdigital la oaie. Lucrrile Sesiunii tiinifice, Institutul Agronomic
Timioara, p. 273-281.
8. Janicki Z., Hrarste A., Slavica A., Konjevi D., Marinovi Z., Stubican D., 2003
- Morphohistological characteristics of the interdigital gland in the roebuck
(Capreolus capreolus L.). Vet. Arhiv 73, 27 - 37
9. Knig E.H., Liebich G.H., 1999 Anatomie der Haussugetiere. Lehrbuch und
Farbatlas fr Studium und Praxis. Band I, Bewegungsapparat Schauttauer
Stuttgart, New York.
10. Knig E.H., Liebich G.H., 1999 Anatomie der Haussugetiere. Lehrbuch und
Farbatlas fr Studium und Praxis. Band II, Organe, Kreislauf-und Nervensystem,
Schauttauer Stuttgart, New York.
11. Liebich H.G., Bck P., Budras K.D., Maierl J., Reese S., 1999 Funktionelle
Histologie der Haussugetiere. Lehrbuch und Farbatlas fr Studium und Praxis.
Schattauer Stuttgard, New York.
12. Mihai M., 1984 Cercetri histologice, citologice, ultrastructurale i morfochimice
asupra glandelorsebacee i sudoripare n decursul ciclului estral i al gestaiei la
ovine. Tez de doctorat, Institutul Agronomic Iai, 1984.
13. incai Mariana, 2000 Histologie veterinar, vol. II, Organe i embriologie. Ed.
Mirton, Timioara.
14. Verde N., Pop T., Manolescu N., 1982 Contribuii la ontogeneza i structura
sinusului interdigital la oaie. Seminarul tiinific "Metode noi de sporire a
produciei la animale" Iai, p. 66-67, 1982.

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OBSERVAII ASUPRA UNOR CAZURI DE FORM


CUTANAT A BOLII LUI MAREK LA BROILERI
OBSERVATION ON SOME CASES OF CUTANEOUS MAREKS DISEASE
AT BROILERS

Cristina BORCIL, S. PACA, M. LAZR


U.S.A.M.V. Iai

Research was made using skin samples from broilers 40 to 47 days


from an intensive raising unit. They were unvaccinated against Mareks
disease, and presented cutaneous tumors 3 and 5 mm in diameter, scattered
on the whole surface of the body.
The histopathologyical exam on horizontal and sagittal sections,
showed underepidermic presence of some lymphoid follicles made of
lymphoid elements and new formatted vessels as well as agglomerations
with muffy aspect around the vessels.
At the feather follicles level was found a light lymphoid hyperplasia
around the external and internal epithelial sheath.
Presence of lymphoid follicles was noticed at the adipous tissue also.
Thus based on the distribution of limphoid elements, it is considered
that the starting point of this hyperplasia is the Skin Associated Lymphoid
Tissue.

Key words: broiler, Marek's disease, lymphoid elements

Boala lui Marek este cea mai important boal limfoproliferativ a ginilor,
produs de virusuri din familia Herpesviridae, subfam. Alphaherpesvirinae.
Vrsta de apariie a bolii la puii nevaccinai este de 35-90 de zile, iar la
psrile vaccinate 114-224 de zile (5).
Transmiterea virusului se face numai orizontal pe cale respiratorie, prin
descuamatele epiteliale ale foliculilor plumiferi.
ntr-o prim faz se produce citoliza limfocitelor B, iar apoi o hiperplazie
reticular.
Boala lui Marek, n forma cronic, are urmtoarele localizri: neural,
ocular i cutanat. Forma acut se caracterizeaz printr-o proliferare nodular
sau difuz, la nivelul ficatului, splinei, rinichilor, cordului, ovarului,
proventriculului i pielii (1).
n localizarea cutanat la broileri se remarc o ngroare a pielii, care apare
cutat, foliculii plumiferi fiind turgesceni i mrii n volum pn la 3mm (3).
Leziunile de ordin histologic ale pielii pot fi grupate n 5 tipuri:
- A: mici agregate de celule limfoide (LCA) ce conin limfocite mici,
limfoblaste i rare figuri mitotice;

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- B: mari agregate de celule limfoide cu limfocite mici, un numr


considerabil de plasmocite i rare figuri mitotice;
- C: agregate limfocitare cu un numr egal de limfocite mici i
limfoblaste;
- D: agregate limfocitare foarte mari n principal coninnd limfoblaste
i doar cteva limfocite mici;
- E: agregate limfocitare mari ce conin aproape exclusiv limfoblaste.
Nodulii de dimensiuni mici de la nivelul foliculilor plumoi corespund
tipurilor B i C asociate cu A, iar nodulii fuzionai, mai mari, corespund tipurilor
D i E.
Studiile au artat c leziunile de tip A le preced pe cele de tip B.
Similitudinea unor leziuni din boala lui Marek cu cele din encefalomielitele i
nevritele alergice ale psrilor sugereaz mecanismul imun de producere a
leziunilor (6).
S-au fcut teste imunohistochimice pentru studierea topografiei proteinelor
fosforilate specifice (PP) ale MVD-1 i genelor lor . PP au fost detectate masiv n
zonele de necroz ale unor limfoame cutanate.
Zonele de necroz de la nivelul pulpei penelor corespund n special
limfoblastelor necrozate (2).
Vrsta de apariie a bolii, polimorfismul celulelor hiperplaziate, prezena
incluziilor n epiteliul foliculilor plumiferi i testul de imunofluorescen a
antigenului de suprafa asociat tumorilor sunt elemente de diagnostic diferenial
al bolii (4).
La puii de carne, conform programelor de supraveghere, profilaxie i
prevenire a transmiterii de boli de la animale la om i protecie a mediului,
vaccinarea nu este obligatorie i n mod curent la noi n ar nu se face.

MATERIAL I METOD

Examenele macroscopice i histopatologice s-au efectuat pe probe de


piele de la 6 broileri, cu vrsta cuprins ntre 42 i 47 zile, nevaccinai, provenii
de la unitatea A.
Fragmentele recoltate au fost fixate n formaldehid soluie apoas 10%,
timp de 5 zile i apoi cu soluie Bouin, timp de 2 zile. S-a utilizat metoda includerii
n parafin, seciunile obinute fiind colorate prin metoda HEA.

REZULTATE I DISCUII

La examinarea necropsic a cadavrelor am observat la nivelul pielii de pe


toat suprafaa corpului, o ngroare a acesteia i o mrire n volum a foliculilor
plumiferi pn la dimensiuni ntre 3 i 5 mm n diametru (Fig. 1, Fig. 2).

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Fig. 1 Fig. 2

La examenul histologic pe seciuni orizontale i sagitale s-a constatat


prezena subepidermic a unor foliculi constituii din elemente limfoide i vase de
neoformaie ct i aglomerri perivasculare cu aspect manonal (Fig. 3, Fig. 4,
Fig. 5, Fig. 6).

Fig. 3 Fig. 4

Fig. 5 Fig. 6

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La nivelul foliculilor plumiferi am constatat, de asemenea, o uoar


hiperplazie limfoid n jurul tecilor epiteliale extern i intern (Fig. 7, Fig. 8, Fig.
9, Fig. 10).

Fig. 7 Fig. 8

Fig. 9 Fig. 10

Prezena foliculilor limfoizi a fost semnalat i la nivelul esutului adipos


din hipoderm (Fig. 11, Fig. 12).

Fig. 11 Fig. 12

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Cercetrile curente au avut ca scop reducerea pierderilor prin sacrificare a


broilerilor. n cercetrile viitoare urmeaz s stabilim vrsta real de apariie a
infeciei i raportul dintre aceasta i pierderile prin confiscare.

CONCLUZII

Se semnaleaz localizarea cutanat a bolii lui Marek la puii broiler,


nevaccinai, n vrst de 40-47 zile.
Macroscopic modificrile constau ntr-o mrire n volum a foliculilor
plumiferi, tradus la nivel structural printr-o proliferare limfoid ce se extinde de
la nivelul dermului pn n esutul conjunctivo-adipos al hipodermului i chiar n
muchii pieloi.
Pe baza distribuiei considerm c punctul de plecare al hiperplaziei
limfoide este esutul limfoid asociat pielii.

BIBIOGRAFIE

1. Ctoi C., 2003 - Diagnostic necropsic veterinar, Ed. Academic Press, Cluj-Napoca,;
2. Cho K.O., Endoh D., Kimura T., Ochiai K., Itakura C., 1997 - Significance of
Mareks disease virus serotype 1 specific phosphorylated proteins in Mareks
disease skin lesions. Avian pathology, 26, 4, 707-720;
3. Cho K.O., Ochiai K., Fujikawa Y., Itakura C., 1997 - Cutaneous lesions in broiler
chickens spontaneously affected with Mareks disease, Avian Pathology, 26, 2,
277-291;
4. Moga Mnzat R., 2005 - Boli virotice i prionice ale animalelor, Ed. Brumar,
Timioara.
5. Paul I., 1996 - Etiomorfopatologie veterinar, Ed. All, Bucureti, 1996.
6. Riddell C. - Avian Histopathology

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ASPECTE LEZIONALE ALE INVAZIILOR PARAZITARE


CU LOCALIZARE BRANHIAL LA CIPRINIDELE DE
CRESCTORIE
M. LAZR1, V. VULPE1, E. BOZ2, L. MIRON1,
B. PSRIN1, O.Z. OPREAN1
1
U.S.A.M.V. Iai
2
Rompescaris SRL Iai

Necropsy exams were made immediately after the extraction from


the natural medium of 64 fishes from the species: carp (Cyprinus carpio),
big head (Aristichthys nobilis) and silver carp (Hypophthalmichthys
molitrix).
The rapid examination of the curettage from the surface of the
brachial lamellas lead to the identification of the fishes from genders
Sinergasillus spp., Dactylogyrus spp. and Trichodina spp.
The histopathological exams revealed gradual irritative lesions,
finalized by hyperplasic mainly lymphocitary of reparation.

Key words: Cyprinidae, gills, parasitical invasion, lesions

Cunoaterea strilor de boal la peti, diagnosticul precoce i aplicarea


corect a msurilor de combatere sunt condiii eseniale pentru asigurarea unei
activiti normale i a unei productiviti crescute n unitile piscicole.
Manifestrile clinice de tip hipoxic apar fie ca urmare a aportului sczut de
oxigen ctre organism, fie datorit leziunilor de la nivelul branhiilor
(ectoparazitoze, toxine endogene sau exogene), fie prin scderea cantitii de
hemoglobin, ca urmare a unor anemii (boli infecioase i parazitare) (1, 2, 4).
Bolile parazitare cu localizare branhial mai des ntlnite sunt: criptobioza
(Cryptobia branchialis), mixosporidioza (Myxobolus pavlovskii, Myxobolus
dispar), dactilogiroza (Dactylogyrus spp.), sinergasiloza (Sinergasilus lieni,
Sinergasilus major), trichodinoza (Trichodina mutabilis, Trichodina nigra,
Trichodina nobilis, Trichodina acuta) (3, 5, 6).

MATERIAL I METOD

Cercetrile noastre au avut ca obiectiv modificrile produse de unii parazii


la nivelul branhiilor, la urmtoarele specii de peti dulcicoli din amenajri piscicole
din judeul Iai: crap (Cyprinus carpio) 20 cazuri, caras (Carassius auratus
gibelio) 6 cazuri, snger (Hypophthalmichtys molitrix) 9 cazuri i novac
(Aristichtys nobilis) 11 cazuri.
Branhiile au fost examinate dup secionarea parial sau total a
operculilor, urmrindu-se culoarea i prezena hemoragiilor ca rezultat al atacului

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paraziilor, prezena unei cantiti excesive de mucus (urmare a iritaiilor


parazitare i a reaciei de aprare a organismului).
S-a efectuat un examen microscopic direct al branhiilor dilacerate cu un ac
spatulat (i izolarea paraziilor) sau a raclatelor de pe branhii.
Pentru realizarea examenului histopatologic s-au extras branhii ntregi de la
petii de talie mic sau poriuni de 1-2 cm din arcul branhial la petii de talie mare
care au fost fixate n formaldehid 10% soluie apoas, incluse n parafin,
secionate la 5 m i colorate cu metodele Hematoxilin Eozin Albastru de
metil (HEA) i Acid periodic Fuxin Schiff (PAS).

REZULTATE I DISCUII

Dup efectuarea investigaiilor epidemiologice i morfopatologice (macro-


i microscopice) au fost diagnosticate trei invazii parazitare cu localizare
branhial la speciile de ciprinide luate n studiu: sinergasiloza, dactilogiroza i
trichodinoza.

Sinergasiloza
Agentul etiologic care produce boala la ciprinidele asiatice (snger, novac)
este Sinergasilus lieni.
S-a constatat c vara invazia parazitar atinge cotele cele mai ridicate, n
iulie-august i afecteaz n general petii n vrst (vara II i vara III).
La examinarea macroscopic, paraziii apar sub forma unor filamente
scurte, albicioase, ncorporate n esutul muscular, de la marginea branhiilor; la
vrful lamelelor branhiale se observ acumulri de mucus translucide (Fig. 1,
Fig. 2).

Fig.1 Hiperplazii branhiale difuze Fig.2 Sinergasillus lieni Armtura bucal.

Examenele histologice au pus n eviden reacii tisulare de grade diferite i


prezena parazitului care dilacereaz lamelele branhiale, hiperplaziile celulelor
clorale i cele limfohistioplasmocitare interlamelare (Fig. 3, Fig. 4, Fig. 5, Fig. 6,
Fig. 7).

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Fig. 3 Lam branhial. Vacuolizarea. Fig. 4 Hiperplazia celulelor clorale. Col.


celulelor clorale. Col. HEA x 400 HEA x 400

Fig.7 Lam branhial.


Fig.5 Sinergasillus lieni. Hiperplazie
Fig.6 Chistizarea spaiilor
Seciune transversal. Saci limfohistioplasmocitar
interlamelare. Col.HEAx100
ovigeri. Col. HEA x 400 interlamelar.
Col. HEA x 400

Dactilogiroza
Aceasta este o parazitoz branhial ce produce pierderi importante la
puietul din fermele ciprinicole i salmonicole.
Agenii etiologici ai dactilogirozei sunt specii ale genului Dactylogyrus.
Parazitul are ca particulariti morfologice patru papile contractile i patru
pete pigmentare la extremitatea cefalic a corpului. La extremitatea posterioar se
afl discul de fixare prevzut cu dou crlige mari centrale, n form de secer i
14 crlige mici, aezate marginal (Fig. 9).
La examenul histologic, se observ la locul de fixare a paraziilor procese
inflamatorii, tumefiere, iar branhiile devin mai groase ducnd la o ngreunare a
circulaiei sngelui i a schimbului de gaze.
ngreunarea circulaiei sanguine nu mai permite hrnirea esutului branhial,
ducnd la moartea acestuia i la apariia unor zone ntinse de necroz la nivelul
lamelelor branhiale (Fig. 10, Fig. 11, Fig. 12, Fig. 13, Fig. 14).
Lezarea esutului branhial conduce la o insuficien respiratorie a petelui
bolnav, petele prezentnd operculele deprtate (Fig. 8).

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Fig. 8 Hiperplazii branhiale nodulare Fig. 9 Dactylogyrus spp. Pete oculare

Fig.10 Denudarea vrfurilor Fig.11 Lamele branhiale. Fig.12 Lame branhiale.


lamelelor branhiale. Col. Epiteliul superficial Hiperplazii la baza lamelelor.
HEA x 100 cuboidizat. Col. HEA x 400 Col. HEA x 400

Fig. 13 Lam branhial. Degenerarea Fig. 14 Lam branhial. Cartilaj degenerat.


cartilajului bazal. Col. HEA x 100 Hiperplazie la baza lamelelor.
Col. HEA x 100

Trichodinoza
Agenii etiologici ai trichodiniozei sunt reprezentai de un grup de ciliate
aparinnd familiei Urceolariidae, cele mai comune fiind speciile genurilor
Trichodina, Trichodinella i Tripartiella (Fig. 16).

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Aspectul acestor ciliate este de clopot mult turtit, de dimensiuni variabile,


pe margini avnd o coroan de cili cu ajutorul creia se deplaseaz pe corpul
gazdei sau de la un pete la altul.
Examinarea microscopic a raclatului profund din zonele afectate permite
identificarea paraziilor.
La examenul histologic, leziunile de la nivelul lamelelor branhiale constau
n descuamarea epiteliului respirator, hemoragii punctiforme, mucus abundent,
necroze branhiale (Fig. 15, Fig. 17, Fig. 18).

Fig. 15 Necroza branhiilor Fig. 16 Trichodina nobilis

Fig. 17 Lam branhial. Congestie bazal. Fig. 18 Necroza lamei branhiale.


Hemoragii interlamelare. Col. HEA x 100 Col. HEA x 100

CONCLUZII

Investigaiile epidemiologice i morfologice, macro- i microscopice


ntreprinse ntr-o ferm piscicol, pe 64 ciprinide din speciile crap, caras, snger
i novac au condus la formularea urmtoarelor concluzii:
1. Examenul necropsic al branhiilor evideniaz necroze i ngrori difuze
sau nodulare pe marginea arcului branhial.

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2. Examenul microscopic extemporaneu al raclatului branhial permite


identificarea agenilor etiologici (Sinergasillus lieni, Dactylogyrus spp.,
Trichodina nobilis).
3. La examinarea microscopic a seciunilor obinute prin metoda includerii
la parafin se observ n general reacii tisulare de rspuns la aciunea iritativ a
paraziilor.
4. Predomin hiperplaziile celulelor clorale i cele limfohistioplasmocitare
interlamelare.
5. Invaziile parazitare prelungite duc la necroze lamelare i depolimerizarea
mucopoliozidelor acide (MPZA) din cartilajul branhial.

BIBLIOGRAFIE

1. Bud I., Diaconescu ., Mudure M., 2004 Creterea crapului i a altor specii, Ed.
Ceres, Bucureti.
2. Grozea A., Bura M., 2002 Crapul: biologie, sisteme de crestere, patologie; Ed. de
Vest, Timisoara.
3. Munteanu G., Bogatu D., 2003 Tratat de ihtiopatologie, Ed. Excelsior Art,
Timisoara.
4. Oel V., Constantin Gh., 1989 Ghid ihtiopatologic pentru piscicultura din Delta
Dunrii, Buletin de Cercetri Piscicole, Supliment II.
5. Rdulescu I., Lustun L., Voican V., 1976 Bolile petilor, Ed. Ceres, Bucureti.
6. Vulpe V., 2002 Investigaii privind epidemiologia, etiologia i morfologia afeciunilor
parazitare la petii din amenajrile sistematice, Tez de Doctorat.

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FORENSIC EXAMINATION IN BITTEN WOUNDS


CAUSED BY DOGS TO MAN
EXPERTIZA MEDICAL LEGAL N PLGILE MUCATE PROVOCATE
DE CINE LA OM

Anca BULU1, C. SCRIPCARU2, O.Z. OPREAN1


1
U.S.A.M.V. Iai
2
I.M.L. Iai

The investigations have as starting point 35 forensic certificates


released by the Legal Medicine Institute (LMI) during 1.01.2003
31.12.2005, in cases of bitten wounds made by dog to people.
The channel of prevention and fighting the main zoonosis
transmissible through dog bite was followed.
Juridical aspects of the approached problems are presented.

Key words: dog, bite, legislation

Prin muctur i zgrietur, cinele poate transmite la om rabia i


pasteureloza (4).
Virusurile rabice aparin Familiei Rhabdoviridae, Ord. Monomegavirales.
Familia e mprit n dou genuri distincte: G. Vesiculovirus include virusuri care
produc stomatitele veziculoase, iar G. Lyssavirus include virusul rabic i virusuri
nrudite cu cel rabic. Prin utilizarea tehnicilor serologice moleculare sunt
recunoscute n prezent 4 serotipuri de lyssavirusuri, cu 6 genotipuri (2).
Transmiterea bolii se produce atunci cnd virusul prezent n saliva unui
animal infectat contamineaz esuturile victimei. Virusul rabic nu poate penetra
tegumentul intact, dar poate ptrunde n organism traversnd mucoasele intacte
(nazal, bucal, anal, conjunctival, a organelor genitale externe). Infecia uman
pe cale aerian este extrem de rar, nsa pare a fi posibil (4).
Mecanismul patogenetic al infeciei rabice const n propagarea centripet a
virusului pe calea filetelor nervoase de la periferie spre neuronii sistemului nervos
central, capacitate denumit de tefan S. Nicolau neuroprobazie. Viteza de
naintare a virusului de-a lungul axonilor este de 3 mm pe or (1).
Diagnosticul se realizeaz n dou etape: izolarea virusului prin inocularea
intracerebral a oarecilor sugari sau nrcai, urmat de testul cu anticorpi
fluoresceni (9).
Pasteurelozele includ o multitudine de entiti morbide produse de diferite
specii din genul Pasteurella spp. la animale i la om. Speciile patogene sunt:
Pasteurella multocida, P. septica (mamifere, om, psri), P. gallicida (pui), P.
dagmatis (cine, pisic, om), P. canis biotip 1 (cine, om), P. canis biotip 2
(viei), P. stomatis (cine, pisic, se transmite la om prin muctur), P. anatis
(rae), P. avium (psri) (4).

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La om, infecia cu Pasteurella multocida e zoonoza cea mai frecvent


raportat, fiind rezultatul mucturilor pisicilor i cinilor infectai. P.
haemolytica a fost de asemeni asociat cu plgile prin muctur de cine.
Infeciile cu P. dagmatis i P. canis biotip 1 au fost asociate cu mucturi i
zgrieturi produse de cine i de alte animale. Dintre aceste specii, P. dagmatis
poate produce i infecii sistemice (1).

MATERIAL I METOD

Investigaiile au fost efectuate la trei obiective (instituii).


Au fost luate n studiu cazurile de muctur de cine la om, pentru care
au fost eliberate certificate medico-legale de ctre Institutul de Medicin Legal
(IML), Iai, pe durata a trei ani: 2003, 2004, 2005.
Cazuistica a fost clasificat pe arii sociale de provenien (Iai i mediul
rural), sexe i vrste.
La Centru Antirabic al Spitalului de Boli Infecto-Contagioase Sf. Cuvioas
Paraschiva Iai a fost luat n studiu numrul de pacieni care figureaz n
evidene n perioada 2003-2005 i schemele de vaccinare preexpunere si
postexpunere.
La Direcia Sanitar Veterinar i pentru Sigurana Alimentelor (DSVSA),
Iai, au fost cercetate registrele de monitorizare a rabiei pe teritoriul judeului Iai,
n care au fost nregistrate 281 probe n perioada 2003-2005.

REZULTATE I DISCUII

La Institutul de Medicin Legal, unde a fost iniiat studiul, n perioada


urmrit au fost eliberate 35 certificate medico-legale, ntr-o uoar progresie
numeric (7 n 2003, 12 n 2004 i 16 n 2005) (Tabel 1).
Tabelul 1
Certificate medico-legale eliberate de IML Iai n perioada 2003-2005

Nr. Cine Om
An crt. Necunoscut Identificat Mediu social Sex Vrst
1 * Rural F 14
2 * Urban F 30
3 * Rural M 62
2003

4 * Rural F 77
5 * Urban M 17
6 * Urban M 38
7 * Rural M 40
8 * Urban F 35
9 * Urban M 58
10 * Rural F 22
2004

11 * Urban M 14
12 * Rural M 72
13 * Rural M 21

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Nr. Cine Om
An crt. Necunoscut Identificat Mediu social Sex Vrst
14 * Urban M 13
15 * Urban M 17
2004

16 * Urban M 8
17 * Rural M 35
18 * Urban F 46
19 * Urban M 12
20 * Rural M 12
21 * Urban F 1
22 * Urban F 53
23 * Urban M 66
24 * Urban M 44
25 * Rural F 50
26 * Urban M 36
27 * Urban F 10
2005

28 * Urban F 47
29 * Rural M 6
30 * Urban M 19
31 * Urban F 46
32 * Rural F 32
33 * Urban M 10
34 * Urban F 29
35 * Urban M 30

n mediul urban (Municipiul Iai) apar mai multe cazuri dect n cel rural
(66% din cazuistic) datorit numrului mare de cini comunitari nesupravegheai
(Diagrama 1).
Diagrama 1
Incidena mucturilor de cine
la om n mediile sociale urban i rural.

Rural
11

Urban
23

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n ceea ce privete sexul victimelor, se constat un plus de 20% al


victimelor brbai (Diagrama 2).
Diagrama 2
Incidena mucturilor de cine
la om n relaie cu sexul victimelor

Femei
Barbati 14
20

Dintre cele 35 cazuri, cele mai numeroase (10) sunt copii i adolesceni cu
vrsta ntre 10 i 20 ani. Au fost mucai doar 3 ceteni n vrst de peste 60 ani
i doar 5 copii n vrst de pn la 10 ani, acetia din urm ieind pe strad mai
ales nsoii de prini (Diagrama 3).
Diagrama 3
Incidena mucturilor de cine la om n relaie cu vrsta

10
9
10
8
7 8 8
6
5
4
3 5
2 3
1
0
0-10 ani 10-20 ani 20-40 ani 40-60 ani >60 ani

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Certificatele medico-legale conin descrierea morfoclinic a leziunilor, data


producerii acestora i necesarul de zile de ngrijiri medicale pn la poteniala
vindecare. Nu sunt menionate date anamnetice cu privire la identificarea i
statusul sanitar al cinilor care produc plgi prin muctur i nici msuri legale
care privesc proprietarii acestora.

n perioada investigat, la Centrul Antirabic din cadrul Spitalului de Boli


Infecioase Iai s-a nregistrat o medie de 60 de pacieni pe lun care au prezentat
mucturi provocate de cini necunoscui sau nevaccinai i n cazul crora s-a
instituit una dintre schemele de tratament. Este de semnalat c numrul de
pacieni variaz considerabil n funcie de perioada anului, incidena mucturilor
vara fiind mai mare dect iarna.
ntre 1.01.2003 si 31.12.2005, n evidenele Centrului Antirabic figureaz
1443 de persoane care au urmat tratament.
Profilaxia pentru rabie sau tratamentul preexpunere se aplic persoanelor cu
risc mare de expunere la infecie (din laboratoare de diagnostic i cercetare care
manipuleaz virusul rabic); se utilizeaz un vaccin antirabic cu virusul WISTAR
RABIES PM/WI 38-1503-3M inactivat i purificat VERORAB.
Tratamentul postexpunere, dup contactul confirmat sau suspectat cu
agentul etiologic, este diferit n funcie de statusul imun al pacientului. La
subiecii neimunizai se administreaz 5 injecii cu vaccin VERORAB n zilele 0,
3, 7, 14 i 28. La subiecii vaccinai n ultimii 5 ani se aplic dou injecii n zilele
0 i 3.
Serul antirabic, obinut din sngele cailor hiperimunizai cu ser antirabic, se
administreaz persoanelor mucate grav de animale turbate sau suspecte de rabie.
Seroterapia este urmat obligatoriu de vaccinoterapie antirabic, care se instituie
n aceeai zi sau n ziua imediat urmtoare.

La Direcia Sanitar Veterinar i pentru Sigurana Alimentelor Iai,


registrele care monitorizeaz rabia pe teritoriul judeului Iai evideniaz 281
probe analizate n perioada 2003-2005. Dintre acestea 212 (75%) sunt probe de la
cini, un singur caz fiind pozitiv, n anul 2004 (Tabel 2, Tabel 3, Tabel 4).
Tabelul 2
Situaia rabiei n perioada 1.01.2003 31.12.2003, pe aria judeului Iai

Nr. total de
Specie Pozitive Negative Improprii
cazuri
Cine 86 0 83 3
Pisic 9 0 9 0
Bovin 1 0 1 0
Ovin 2 0 2 0
Vulpe 7 0 6 1

Total cazuri: 106 (negative)

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Tabelul 3
Situaia rabiei n perioada 1.01.2004 31.12.2004 pe aria judeului Iai

Nr. total de
Specie Pozitive Negative Improprii
cazuri
Cine 87 0 87 0
Pisic 11 0 11 0
Bovin 1 0 1 0
Ovin 4 0 4 0
Caprin 2 0 2 0
Vulpe 8 4 4 0
Dihor 2 0 2 0

Total: 115 cazuri, din care 4 pozitive, la vulpe: 2 vulpi din localitatea Schitu Duca, o
vulpe din satul Ciorleti i o vulpe din pdurea de la Dobrov.

Tabelul 4
Situaia rabiei n perioada 1.01.2005 31.12.2005 pe aria judeului Iai

Nr. total de
Specie Pozitive Negative Improprii
cazuri
Cine 39 1 38 0
Pisic 5 1 4 0
Bovin 2 0 2 0
Vulpe 12 1 11 0

Total: 60 de cazuri, din care 3 pozitive: un cine din comuna Mirosloveti, satul Merce,
o pisic din comuna Bivolari i o vulpe din localitatea Schitu Duca.

Din totalul de 212 probe de la cini suspeci, doar 7 au provenit de la cini


necunoscui, restul fiind, n majoritate, cini cu stpn, provenii din mediul rural
si nevaccinai.
Este de remarcat faptul c n judeul Iai, n perioada 2000-2002, au fost
diagnosticate doua cazuri mortale de rabie la om, ceea ce atrage atenia asupra
unei foarte stricte supravegheri a bolii n zon.
n Laboratorul Judeean Sanitar Veterinar Iai se fac examene complete i
prin tehnici moderne de diagnostic al turbrii: necropsii, examene histopatologice,
proba biologic, identificarea imunochimic a virusului rabic.

Cadrul legal care reglementeaz situaia cinilor din Romnia se refer in


principal la animalele cu stpn: legea nr. 61/27.09.1991 pentru sancionarea
faptelor de nclcare a unor norme de convieuire social, a ordinii i linitii
publice (10) i Ordonana de Urgena nr. 55/30.04.2002 privind regimul de
deinere a cinilor periculoi sau agresivi (7). Msurile legale adoptate n vederea
rezolvrii problemei cinilor comunitari (care provoac de altfel majoritatea
accidentelor) nu au dat pn n prezent rezultatele scontate (5).

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CONCLUZII

Investigaiile efectuate cu privire la importana epidemiologic i impactul


sanitar i social al mucturilor de cine la om n trei obiective (instituii):
Institutul de Medicin Legal, Direcia Sanitar Veterinar i pentru Sigurana
Alimentelor i Spitalul de Boli Infecto-Contagioase din Municipiul Iai, relev o
discontinuitate n colaborarea celor trei instituii cu privire la gravitatea
mucturilor de cine la om i la importana acestora n meninerea sntii
publice.

BIBLIOGRAFIE

1. Carp Crare M., 2001 - Microbiologie veterinar Virusologie; Ed. Venus, Iai;
2. Enache T., Paul I., Popescu O., Stnescu V., Iordache I., 1997 - Medicina legal
veterinar, Vol. I; Ed. ALL, Bucureti;
3. Oprean O.Z., 2002 - Diagnosticul necropsic la carnivorele de companie; Ed. Ion
Ionescu de la Brad, Iai;
4. Palmer S.R., Lord Soulsby, Simpson D.I.H., 2005 - Zoonoze; Ed. tiine Medicale,
Bucureti;
5. Scripcaru C., Scripcaru Gh., Astarastoae V., 2005 - Medicina legal pentru juriti;
Ed. Polirom, Iai;
6. X X X - Legea sanitar veterinar nr. 75/1991;
7. X X X - Legea nr. 61 din 27 septembrie 1991 pentru sancionarea faptelor de
nclcare a unor norme de convieuire social, a ordinii i linitii publice;
8. X X X - Ordonana de Urgen nr. 55 din 30 aprilie 2002 privind regimul de deinere
al cinilor periculoi sau agresivi;
9. X X X - Ordinul 156/1999 pentru aprobarea Normei sanitare veterinare privind
anunarea, declararea i notificarea unor boli transmisibile ale animalelor;
10. X X X - Manual OIE/2004

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LEZIUNILE HIPERPLAZICE/DISPLAZICE ADIACENTE


TUMORILOR MAMARE MALIGNE
HIPERPLASTIC/DISPLASTIC LESIONS IN TISSUE SURROUNDING MAMMARY
MALIGN TUMORS

A. GAL, A.I. BABA, C. CTOI,


Sorina IACOB, E. DOMBAY, I. RUS

The aim of the study was to notify the displastic/hiperplastic


transformations from mammary tissue surrounding mammary malign
neoformations. It is known that displastic/hiperplastic mammary lesions
could be preneoplastical stages. There was established, histologically, the
types of mammary neoplasms, the lesion character and
displastic/hiperplastic lesions from adjacent area. There were studied 76
mammary tumors, 15 of them presented displastic/hiperplastic changes in
mammary parenchyma adjacent to malign areas. The tumors were from 14
bitches and 1 female cat. The most affected breed were Caniche (6 cases),
Cocker (3 cases), Stray dog (2 cases), Hungarian vijla, Pitt/Bull, German
dog (1 case). The age varied from 5-12 years; the majority displastic
lesions were noticed in 10 years old dog, with a mean incidence of 8,3
years. The mammas with such kind of lesions were: M5 46,1%, M4
38,4%, M3 7,6%, M2 7,6%. The tumor types with displastic/hiperplastic
lesions were: complex tubular carcinoma (5 cases), complex tubulo-
papillary carcinoma (2 cases), simple tubulo-papillary carcinoma (2 cases),
solid carcinoma (1 case), complex carcinoma in benign mixed tumor (3
cases), in situ carcinoma (1 case), mammary sarcoma (1 case). The most
important displastical lesions were lobular hyperplasia such us typical
epitheliosis (12 cases), atypical epitheliosis (1 case) and simple/complex
adenosis (6 cases); ductal hyperplasia such us: tipical epiteliosis (4 cases),
papilomatosis (1 case). Displastic lesions were: cyst displasia (8 cases) and
ductal ectasia (3 cases). In conclusion, displastic/hiperplastic
transformations are quite frequent encountered in pericarcinomatous areas.
These lesions increase the tumor risk development of 2-5 times, especially
typical and atypical epitheliosis. As a recommendation, surgical therapy
must involve excision of displastic/hiperplastic mammary tissue too, not
only mammary tumor.

Key words: bitch, mammary gland, tumors

MATERIAL I METOD

Materialul prezentei lucrri a fost constituit din cadavrele aduse pentru


examenul necropsic la FMV Cluj Napoca, precum i biopsiile tumorale provenite
de la cabinetele veterinare sau de la disciplina de chirurgie. Examenul necropsic
a constat n aprecierea dimensiunii, consistenei, aspectului pe seciune,

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localizarea neoformaiunii, integritatea limfonodulilor regionali, urmnd apoi


recoltarea de fragmente pentru preparatele histologice. Fragmentele au fost
recoltate din zonele periferice ale tumorii, din 2-3 pri ale acesteia, evitndu-se
ns zonele necrotice, necrotico-purulente sau chistice. Poriunile recoltate au
vizat ntotdeauna i zonele marginale ale tumorii, n vederea prevelrii i de esut
mamar limitrof. Fragmentele prelevate au fost fixate n formol neutru 10%,
prelucrarea fcndu-se ulterior prin technica la parafin. Seciunile au fost
colorate prin technica Tricrom Masson i hematoxilin-eozin.

REZULTATE OBINUTE

Cazul 1: Cea, rasa Cocker, 10 ani.


Anamnez i aspecte macroscopice: femela prezenta trei formaiuni
tumorale mamare, localizate la nivelul lantului mamar stng, respectiv n mamela
abdominal posterioar i n mamela inghinal. Una dintre tumori avea
aproximativ 1,5 cm diametru iar celelalte dou erau mici, globuloase, ct un bob
de mazre (0,5 cm diametru).
Aspecte histologice: formaiune tumoral de natur malign caracterizat
de structuri tubulare care de obicei delimiteaz un lumen, unele structuri tubulare
fiind chistizate, intraluminal evideniindu-se un coninut cu tinctorialitate
variabil. Celulele tumorale prezint nuclei mari, cu nucleoli evideni, uneori
chiar monstruoi. Se poate evidenia un pleiomorfism celular moderat, unele
celulele din structurile tubulare fiind de dimensiuni mici, cu nuclei relativ
uniformi, existnd ns i structuri tubulare n care se poate evidenia un pregnant
polimorfism celular. Pe alocuri se pot evidenia poliferri tumorale cu celule
mioepiteliale, precum i structuri ductale cu creteri papilifere.
Diagnostic histologic: carcinom tubular complex.
Leziuni displazice/hiperplazice: hiperplazie lobular tipul adenozis (o
proliferare netumoral a ductelor intralobulare), la nivelul unui singur lobul
mamar observndu-se zone cu hiperplazie epitelial acinar, cu celule tinere i
nuclei evideni.

Cazul 2: Cea, rasa Caniche, 12 ani.


Anamnez i aspecte macroscopice: formaiune tumoral mamar de
aproximativ 2 cm diametru, limfonodurile inghinale fiind hipertrofiate, dense la
palpare i mobile. Formaiunea tumoral a aprut n urm cu 6-7 luni, n urm cu
un an ceaua fiind operat pentru o formaiune proliferativ de la nivel vaginal.
Tumora mamar nu era aderent la piele, fiind ns aderent la mamela inghinal.
Glanda mamara inghinal opus prezenta o formaiune calcificat ct un bob de
mazre, de sub 1 cm diametru. Femela a fost sterilizat i avea endometrit
hiperplastic i chiti ovarieni.
Aspecte histologice: formaiune tumoral malign caracterizat de structuri
tubulare, reprezentate din celule epiteliale n multistrat, cu numeroase protruzii

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papilare i/sau micropapilare intraductale. Din loc n loc se observ proliferri de


celule mioepiteliale ntr-o mas de mucus care n unele zone are o tent uor
bazofil. Stroma conjunctiv este abundent, pe alocuri infiltrat cu
limfohistiocite.
Diagnostic histologic: carcinom tubulo-papilar complex.
Leziuni displazice/hiperplazice: se pot evidenia lobuli hipertrofiai cu
hiperplazie lobular de tipul adenozis simplu sau lobuli hipertrofiai datorit
chisturilor acinare. Numeroi lobuli mamari sunt transformai n ntregime n
structuri cu aspect chistic. Unele ducte extralobulare sunt i ele chistizate (ectazii
ductale extralobulare). La nivelul unor acini mamari se poate observa o
hiperplazie acinar netumoral - epiteliozis tipic.

Cazul 3: Cea, rasa Caniche, 9 ani.


Anamnez i aspecte macroscopice: formaiune tumoral mamar de
mrimea unui ou de porumbel, cu aspect globulos, consisten moale, neoplasmul
fiind aderent la piele, cu o perioad evolutiv de aproximativ 5 luni.
Aspecte histologice: tumor mamar malign caracterizat printr-o
poliferare tumoral mixt epitelial i mioepitelial. Celulele epiteliale sunt
dispuse sub form de tubi, la nivelul acestora existnd mai mult de dou rnduri
de celule tumorale, uneori chiar cu formarea de papile ductale. Componenta
mioepitelial este bine reprezentat sub form de noduli care de multe ori au
tendina de a forma esut cartilaginos. Stroma conjunctiv este groas cu fibrocite
i fibre groase de colagen, pe alocuri aceasta fiind infiltrat de limfocite, histiocite
i macrofage.
Diagnostic histologic: carcinom tubular complex.
Leziuni displazice/hiperplazice: parenchimul mamar adiacent tumorii
prezint leziuni displazice i hiperplazice, evideniindu-se chisturi acinare precum
i hiperplazii lobulare cu aspectul de epiteliozis papilar tipic, aspect ntlnit att la
nivelul acinilor ct si la nivelul unui duct intralobular. Pe lang cele prezentate
anterior, am evideniat i o hiperplazie ductal de tipul epiteliozis papilar tipic
ntr-un canal extralobular.

Cazul 4: Cea, rasa Pitt-bull, 5 ani.


Anamnez i aspecte macroscopice: formaiune tumoral mamar de
mrimea unei alune, globuloas, localizat ntr-o gland mamar pectoral,
tumora fiind dens i aderent la parenchimul mamar adiacent. Timpul de evoluie
a tumorii a fost de cteva sptmni.
Aspecte histologice: neoformaiune tumoral malign, caracterizat prin
poliferri cu aspect tubular i papilar intraductal. Proliferrile papilifere
intraductale prezint sau nu o strom conjunctivo-vascular de susinere, n unele
ducte papilele formnd chiar puni transversale. esutul conjunctiv este cu aspect

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tnr, nucleii fiind mari, evideni, cu nucleoli proemineni. n masa tumorii se pot
ntlni hemoragii i focare necrotice.
Diagnostic histologic: carcinom tubulo-papilifer simplu.
Leziuni displazice/hiperplazice: n jurul tumorii se pot evidenia lobuli
hiperplaziai cu aspect de adenozis tipul simplu i tipul complex, la nivelul
ductelor intralobulare i chiar la nivelul acinilor observndu-se hiperplazii de tipul
epiteliozis atipic.

Cazul 5: Cea, rasa Caniche, 6 ani.


Anamnez i aspecte macroscopice: formaiune tumoral localizat n
mamela abdominal anterioar pe lanul mamar stng, dimensiunea fiind de
aproximativ 1 cm, respectiv ct un bob de mazre. Tumoara era bine circumscris
i neaderent la esuturile din jur.
Aspecte histologice: neoformaiune tumoral cu un caracter malign
caracterizat printr-o proliferare a celulelor epiteliale, n general cu aspect solid
sau pe alocuri tubular, cu un epiteliu n multistrat care umple lumenul tubular.
Celulele tumorale sunt agresive cu nucleii mari, nucleoli evideni, supranumerari,
observndu-se mitoze i focare de necroz n masa tumorii. Stroma conjunctiv
este discret, pe alocuri fibroblastele avnd un aspect tnr cu nuclei i nucleoli
mari. n stroma conjunctiv se ntlnete un abundent infiltrat limfo-histiocitar i
macrofagic.
Diagnostic histologic: carcinom solid.
Leziuni displazice/hiperplazice: n lobulii mamari peritumorali se observ
displazie chistic acinar precum i atrofia de compresiune a lobulilor mamari iar
la nivelul ductelor intralobulare se observ i o hiperplazie lobular epitelial
tipic.

Cazul 6: Cea, rasa Caniche, 6 ani.


Anamnez i aspecte macroscopice: prezena unui nodul n lanul mamar
drept, la nivelul mamelei inghinale. Dimensiunea nodulului era de 1,5 cm
diametru, consistena fiind crmoas. Neoformaiunea a fost extirpat mpreun
cu mamela inghinal, efectundu-se n acelai timp i ovario-histerectomia.
Aspecte histologice: neoformaiune tumoral malign, reprezentat printr-
un caracter proliferativ de tip papilifer, aspect ntlnit i n unii lobuli mamari, att
la nivelul ductelor intralobulare ct i la nivelul acinilor mamari. Datorit pstrrii
n unele zone a structurii lobulare, precum i datorit evidenierii n masa de
proliferare a microfocarelor de necroz, se poate zice c avem de-a face cu focare
de carcinom in situ i nu cu noduli hiperplazici. Stroma conjunctiv este
abundent, reprezentat de un esut conjunctiv matur, pe alocuri cu infiltrat
mononuclear.
Diagnostic histologic: carcinom tubulo-papilar simplu.

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Leziuni displazice/hiperplazice: la nivelul unui duct galactofor se observ


structuri cu aspect de papilom. Pe lng lobulii transformai tumoral se pot
observa i lobuli nemalignizai hiperplaziai, cu aspect de adenozis simplu. La
nivelul unor acini se pot evidenia focare de hiperplazie epitelial lobular
respectiv epiteliozis tipic.

Cazul 7: Cea, rasa Vijla maghiar, 5 ani.


Anamnez i aspecte macroscopice: formaiuni tumorale multiple cu aspect
slninos, culoare alb-sidefie, localizate la nivelul mai multor mamele. Unii noduli
tumorali au aspect chistic cu coninut hemoragico-necrotic.
Aspecte histologice: neoformaiune tumoral de natur malign,
caracterizat prin proliferri epiteliale de tip tubular, n masa tumorii existnd i o
component tumoral benign, respectiv un esut cartilaginos care pe alocuri
prezint focare de mineralizare. ntre structurile tubulare se pot evidenia att
celule mioepiteliale proliferate sub form de noduli, precum i o strom
conjunctiv reprezentat de un esut conjunctiv tnr. Prezena unor microfocare
necrotice n masa tumorii denot o leziune malign.
Diagnostic histologic: carcinom complex n tumor mixt benign.
Leziuni displazice/hiperplazice: n jurul tumorii, parenchimul mamar rmas
prezint o hiperplazie lobular netumoral de tipul adenozis simplu. La nivelul
unor acini mamari se pot observa att hiperplazii epiteliale tipice (epiteliosis
tipic), ct i hiperplazii ductale epiteliale la nivelul ductelor extralobulare. Unii
acini de la nivelul unor lobuli sunt chistizai.

Cazul 8: Cea, rasa Caniche, 9 ani.


Anamnez i aspecte macroscopice: femela prezenta trei formaiuni
nodulare de diferite dimensiuni n parenchimul mamelelor inghinale, un nodul
fiind ct un ou de gin iar celelalte dou formaiuni avnd dimensiunea unei
alune. Pe seciune, nodulul cel mai mare exprima un material purulent,
formaiunea fiind bine delimitat de o capsul groas.
Aspecte histologice: proliferare tumoral de natur malign reprezentat de
structuri tubulare discrete, celulele avnd n general un aspect fusiform, alungit
sau stelat, unele structuri tubulare fiind chistizate i cu un coninut oxifil bogat n
neutrofile. Pe alocuri se observ focare de metaplazie cartilaginoas, uor
bazofile. Pe zone ntinse ntlnim proliferri mioepiteliocitare cu aspect polimorf
iar stroma conjunctiv peritubular prezint macrofage cu un pigment brun.
Diagnostic histologic: carcinom complex n tumor mixt benign.
Leziuni displazice/hiperplazice: displazie chistic acinar i hiperplazie
lobular cu aspect de epiteliozis tipic acinar, tipul papilar.

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Cazul 9: Cea, rasa Dog german, 8,5 ani.


Anamnez i aspecte macroscopice: formaiune tumoral mamar cu
dimensiunea de 1 cm, de form alungit, bine delimitat de esuturile din jur, cu o
uoar reacie pe cordonul limfatic posterior. Formaiunea tumoral era localizat
pe lanul mamar drept, la nivelul mamelei abdominale posterioare. Ceaua a
nscut o singur dat la vrsta 4 ani i, de atunci, a prezentat de dou ori lactaie
fals.
Aspecte histologice: formaiune tumoral malign caracterizat de un
aspect histologic tubulo-chistic, la nivelul unor chisturi evideniindu-se structuri
micropapilare cu sau fr strom conjunctiv de susinere. Structurile papilifere se
pot observa n majoritatea tubilor. Epiteliul tubular este n multistrat, cu nuclei
mari, veziculoi, nucleoli evideni, n masa tumorii ntlnindu-se i zone de
proliferare mioepitelial, precum i focare de hemoragie i necroz. Stroma
conjunctiv este n general, discret pe alocuri evideniindu-se i infiltrat limfo-
histiocitar.
Diagnostic histologic: carcinom tubulo-papilar complex.
Leziuni displazice/hiperplazice: la nivelul unui lobul mamar s-au evideniat
displazii de tipul chisturilor intralobulare. ntr-o alt seciune histologic la
nivelul unui lobul mamar peritumoral se pot observa leziuni hiperplazice lobulare
de tipul epiteliozis tipic, chiar cu formarea la nivelul unor acini de puni celulare.

Cazul 10: Cea, metis, 10 ani.


Anamnez i aspecte macroscopice: formaiune tumoral mamar
localizat pe lanul mamar drept, ntre M4 i M5, de 5 cm diametru. n M4 era
prezent o alt tumor de dimensiuni mai mici, nodulul fiind sub tensiune,
neaderent la piele i la parenchimul mamar adiacent. Formaiunea localizat ntre
cele dou mamele era un chist purulent. Femela prezenta la nivel vulvar un
material purulent, cenuiu.
Aspecte histologice: formaiune tumoral malign caracterizat prin
proliferri tumorale cu aspect tubular i cu o pregnant proliferare
mioepiteliocitar. Celulele tumorale epiteliale formeaz structuri tubulare, cu
prezena mai multor straturi de celule care formeaz n unii tubi structuri papilare.
La nivelul unor tubi proliferativi componenta epitelial este slab reprezentat,
predominnd componenta mioepitelial. De asemenea, n masa tumorii se observ
o strom conjunctiv groas, abundent, n care se ntlnete pe alocuri infiltratul
limfo-histiocitar sub form de noduli.
Diagnostic histologic: carcinom tubular complex.
Leziuni displazice/hiperplazice: lobulii mamari prezeni peritumoral sufer
o atrofie de compresiune, la nivelul acestora evideniindu-se aspecte de
hiperplazie lobular, respectiv epiteliozis tipic n unii lobuli i atipic n ali lobuli.

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Cazul 11: Cea, rasa Cocker, 7 ani.


Anamnez i aspecte macroscopice: formaiune tumoral mamar
localizat pe lanul mamar drept, n glanda mamar inghinal.
Aspecte histologice: neoformaiune de natur malign caracterizat de
structuri tubulare cu o proliferare epitelial i mioepitelial, n lumenul unor tubi
evideniindu-se uneori un material uor bazofil, n general invadat de granulocite
neutrofile. Unii tubi sunt destini cu aspect chistic, prezentnd un coninut
purulent. Celulele tumorale epiteliale sunt polimorfe, cu nuclei mari, nucleoli
evideni. Mioepiteliocitele ntlnite n unii tubi au aspect stelat, cu nuclei n
general coluroi. Tumora prezint o strom conjunctiv abundent cu fibre
groase de colagen, infiltrat pe alocuri cu celule histiocitare.
Diagnostic histologic: carcinom tubular complex.
Leziuni displazice/hiperplazice: la nivelul unor acini mamari se pot observa
structuri displazice de natur chistic.

Cazul 12: Pisic, ras European, 10 ani.


Anamnez i aspecte macroscopice: formaiune nodular mamar cu aspect
chistic, consisten moale, cu chiti multipli de cca. 1-2 mm, ce prezint un lichid
citrin, limpede. Formaiunea a aprut n urm cu un an pe lanul mamar stng, la
nivelul mamelei abdominale posterioare.
Aspecte histologice: formaiune tumoral malign caracterizat printr-o
proliferare tumoral intralobular, lobulul afectat fiind mult destins, datorit
proliferrii intraacinare i intraductale a celulelor epiteliale. Acestea sunt celule
mari i polimorfe, cu nuclei mari, citoplasm slab colorat, nucleoli evideni.
Datorit prezenei microfocarelor de necroz n zonele de proliferare, avem de-a
face cu o leziune malign, respectiv cu un carcinom in situ.
Diagnostic histologic: carcinom lobular in situ.
Leziuni displazice/hiperplazice: lobulii mamari netransformai tumoral sunt
mrii n volum, cu aspect de hiperplazie lobular tipul adenozis simplu, iar la
nivelul canalelor intralobulare se observ hiperplazii de tipul epiteliozis cu celule
bine difereniate care umplu lumenul ductului mamar. n ductele extralobulare se
observ ectazii ductale, precum i hiperplazii epiteliale tipice.

Cazul 13: Cea, metis, 10 ani.


Anamnez i aspecte macroscopice: ceaua a fost sterilizat n urm cu 2
ani. Tumora mamar a aprut n decursul ultimului an, avnd dimensiuni mari (ct
o nuc de cocos) i o consisten crnoas la palpare.
Aspecte histologice: neoformaiune tumoral de natur malign cu un
caracter sarcomatos, caracterizat printr-o proliferare cu celule fusiforme sau cu
aspect dendritic, care pe alocuri formeaz o matrice proteic parial mineralizat,
cu tendin de a forma lame cartilaginoase sau osoase.
Diagnostic histologic: sarcom mamar.

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Leziuni displazice/hiperplazice: n esutul conjunctiv din jurul tumorii se


poate evidenia la nivelul unui lobul, att n ductele lobulare ct i la nivel acinar,
hiperplazii lobulare cu aspect papilar, celulele fiind bine difereniate, diagnosticul
fiind de epiteliosis tipic.

Cazul 14: Cea, rasa Cocker, 8 ani.


Anamnez i aspecte macroscopice: ceaua a fost sterilizat n urm cu 3
ani, tot atunci efectundu-se i ablaia mamelei inghinale drepte. n urm cu 2 ani
a aprut o alt tumor la nivelul mamelei M4 de la nivelul aceluiai lan mamar,
dimensiunea fiind de aproximativ 1 cm diametru.
Aspecte histologice: neoformaiune tumoral malign caracterizat att prin
proliferri epiteliale cu aspect tubular ct i prin proliferri mioepiteliale.
Structurile tubulare n general au un lumen discret, celulele tumorale n cele mai
multe cazuri umplnd lumenul tubular, nucleii fiind mari, veziculoi, nucleolii
supranumerari i evideni. Celulele mioepiteliale proliferate au un aspect nodular,
fiind delimitate de o strom conjunctiv fin. n unele zone ale tumorii se
ntlnesc hemoragii i infiltrat macrofagic abundent, celulele prezentnd un
pigment brun verzui n citoplasm.
Diagnostic histologic: carcinom tubular complex.
Leziuni displazice/hiperplazice: la nivelul parenchimului mamar din
vecintatea tumorii, respectiv n ductele extralobulare, se observ hiperplazii
ductale cu aspect papilifer fr atipii celulare (epiteliozis papilar ductal tipic), iar
n stroma conjunctiv peritumoral se observ hemoragii.

Cazul 15: Cea, rasa Caniche, 10 ani.


Anamnez i aspecte macroscopice: tumor mamar recidivat. Prima
ablaie n urm cu 1an.
Aspecte histologice: formaiune tumoral de natur malign caracterizat
prin multiple formaiuni cu aspect chistic, cu prezena unor protruzii epiteliale
intrachistice, papilele prezentnd n general strom conjunctivo-vascular de
susinere. n unele formaiuni chistice se poate observa un material oxifil cu
macrofage i cristale de colesterol. Stroma conjunctiv a tumorii este abundent,
slab celularizat, cu fibre groase de colagen, proliferrile epiteliale avnd un
aspect tubular, chiar cu prezena de micropapile. De asemenea se pot evidenia la
nivelul stromei conjunctive zone de calcificare intens bazofile.
Diagnostic histologic: carcinom papilar chistic n tumor mixt benign.
Leziuni displazice/hiperplazice: la nivelul unui lobul mamar se pot observa
hiperplazii epiteliale cu aspect tipic (epiteliozis ductal papilar tipic), proliferrile
fiind cu un aspect digitiform de tip punte la nivelul unui duct intralobular. Ca
leziuni displazice secundare, ntlnim chisturi i ectazii ductale intra i
extralobulare.

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DISCUII
Tabelul 1.
Leziuni displazice/hiperplazice mamare diagnosticate la periferia
unor neoplasme maligne mamare

Nr. Vrsta Mamel Leziuni


Rasa Castrare Diagnostic
caz (ani) afectat displazice/hiperplazice
1 Cocker 10 nu M4-M5S - carcinom -adenozis
tubular complex
2 Caniche 12 nu M5 - carcinom -adenozis simplu
tubulo-papilar -ectazie ductal
complex -displazie chistic
-epiteliosis tipic
3 Caniche 9 nu - - carcinom -displazie chistic
tubular complex -epiteliosis papilar tipic
-epiteliosis ductal
papilar tipic
4 Pitt-Bull 5 nu M2 - carcinom -adenozis simplu i
tubulo-papilifer complex
simplu -epiteliosis atipic
5 Caniche 6 nu M3S - carcinom solid -displazie chistic
-epiteliosis tipic
6 Caniche 6 nu M5D - carcinom -papilomatoz
tubulo-papilar -adenozis simplu
simplu -epiteliosis tipic
7 Vijla 5 nu - - carcinom -adenozis simplu
maghiar complex n TMB -epiteliosis ductal tipic
-displazie chistic
8 Caniche 9 nu M5 - carcinom -displazie chistic
complex n TMB -epiteliosis papilar tipic

9 Dog 8.5 nu M4D - carcinom -displazie chistic


german tubulo-papilar -epiteliosis tipic
complex
10 R. c. 10 nu M4-M5D - carcinom -epiteliosis tipic
tubular complex -epiteliosis atipic
11 Cocker 7 nu M5D - carcinom -displazie chistic
tubular complex
12 Pisic 10 nu M4S - carcinom in -adenozis simplu
situ -epiteliosis tipic
-ectazie ductal
-epiteliosis ductal tipic
13 R. c. 10 da - - sarcom -epiteliosis tipic
mamar
14 Cocker 8 da M4D - carcinom -epiteliosis ductal
tubular complex papilar tipic
15 Caniche 10 nu - - carcinom -epiteliosis ductal
papilar chistic n papilar tipic
TMB -displazie chistic
-ectazie ductal

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Tumorile mamare la cea sunt hormonodependente iar riscul de apariie


crete dup fiecare ciclu estral. Celulele tumorale mamare, fie c sunt benigne, fie
c sunt maligne, au receptori pentru estrogeni i progesteron, ceea ce provoac
hipertrofia parenchimului mamar, fapt evident dup estrus. Incidena mai ridicat
a tumorilor n perechile posterioare ale mamelelor, se coreleaz cu volumul mai
mare al acestora i secreia mai abundent n perioada de lactaie.
n studiu a fost luat un numr de 76 de formaiuni tumorale mamare, din
acestea fiind reinute, datorit evidenierii histopatologice de leziuni
displazice/hiperplazice n esutul mamar adiacent, un numr de 15 tumori de
natur malign. Cele 15 cazuri cu neoplasme mamare au provenit de la 14 cele
de diferite rase i vrste, precum i de la o pisic din rasa european.
n ceea ce privete rasele de cini la care s-au ntlnit displaziile mamare, s-
a observat o inciden crescut a acestor leziuni la rasa Caniche (6 cazuri), Cocker
(3 cazuri), Rasa comun (2 cazuri) i Vijla maghiar, Pitt-Bull, Dog german (cte
1 caz).
Cei 15 subieci au avut vrste cuprinse ntre 5 i 12 ani, cele mai multe
leziuni displazice fiind ntlnite la femele n vrs de 10 ani (5 cazuri), media
fiind de 8,3 ani.
Mamelele la nivelul crora s-au evideniat cele mai multe
hiperplazii/displazii au fost: M5 46,1%, M4 38,4%, M3 7,6% i M2 7,6%.
Referitor la tipurile de neoplasme n jurul crora s-au ntlnit leziuni
displazice, acestea au fost: carcinom tubular complex - 5 cazuri, carcinom tubulo-
papilar complex - 2 cazuri, carcinom tubulo-papilar simplu - 2 cazuri, carcinom
solid - 1 caz, carcinom complex n tumor mixt benign - 3 cazuri, carcinom
lobular in situ -1 caz, sarcom mamar - 1 caz. Rezultatele investigaiilor noastre
arat c leziunile displazice/hiperplazice mamare peritumorale sunt frecvent
ntlnite, multe din acestea, dup cum s-a observat n partea bibliografic a
lucrrii, putnd reprezenta de fapt leziuni preneoplazice. Unele leziuni cum ar fi
hiperplaziile lobulare sau ductale de tipul epiteliozis atipic sau adenozisul, de
multe ori pot duce la erori de diagnostic. Din aceast cauz sunt necesare
elemente de diagnostic diferenial al hiperplaziilor fa de leziunile maligne
carcinomatoase.
Investigaiile histologice efectuate de ctre noi au demonstrat necesitatea
excizrii i recoltrii esutului mamar adiacent tumorii datorit incidenei ridicate
a acestor leziuni peritumorale hiperplazice, din care se pot dezvolta ulterior
neoplasme mamare. Principalele leziuni displazice evideniate au fost
hiperplaziile lobulare, iar din acestea cele mai numeroase au fost leziunile de
epitelioz tipic la 12 cazuri, iar la 1 caz s-a observat epiteliozisul slab difereniat
denumit i epiteliozis atipic. Acesta din urm este foarte greu de difereniat de
carcinomul ductal in situ. Hiperplazia lobular de tip adenozis simplu i complex
a fost ntlnit la 6 din cazuri. Tot leziuni hiperplazice se pot ntlni i la nivel
extralobular, respectiv la nivelul ductelor extralobulare, fiind ntlnite i aici

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modificri ca epiteliozisul tipic la 4 cazuri i leziuni de papilomatoz la 1 caz. Ca


leziuni displazice, cele mai numeroase au fost chisturile mamare lobulare la 8
cazuri, iar la 3 cazuri s-a ntlnit ectazia ductal. Aceste leziuni pot fi confundate
cu carcinoamele chistice mamare (destul de rar ntlnite), existnd anumite criterii
de difereniere a celor dou tipuri lezionale, cele dou tipuri lezionale necesit un
tratament complet diferit.
Studiul efectuat de noi indic faptul c leziunile displazice/hiperplazice
sunt frecvent ntlnite n esutul mamar adiacent tumorii, iar extirparea
neoplasmului cu nendeprtarea esutului adiacent preneoplazic, poate duce la o
recidiv tumoral nu din cauza rezeciei incomplete a tumorii ci din cauza acestor
hiperplazii care se pot maligniza. O alt problem ar fi diferenierea leziunilor
displazice/hiperplazice de cele tumorale. Astfel trebuie difereniat hiperplazia
epitelial de carcinomul ductal bine difereniat; hiperplaziile papilare de
carcinomul papilar; mastopatia fibrochistic de carcinomul chistic; proliferrile
ductale atipice de carcinomul in situ, hiperplazia hipersecretoare chistic de
carcinomul hipersecretor chistic, adenozisul microglandular de carcinomul
tubular, adenozisul sclerozat de carcinomul mamar, hiperplazia lobular atipic
(ALH) de LCIS, hiperplazia ductal atipic de carcinomul ductal in situ (1, 3, 4,
6, 8, 10, 11).
Anumite leziuni displazice/hiperplazice cresc foarte mult riscul apariiei de
tumori mamare, astfel adenozisul i adenozisul sclerozat, cicatricea radial,
hiperplazia ductal epitelial tipic i papilomatoza cresc riscul apariiei
carcinoamelor de 1,5-2 ori, iar hiperplazia ductal i lobular atipic (ADH i
ALH) cresc riscul dezvoltrii tumorilor maligne de 4-5 ori (9). Stabilirea tipului i
subtipului de neoplasm mamar, precum i a leziunilor displazice/hiperplazice
mamare are o importan predictiv deosebit, deoarece n funcie de aceast
clasificare histologic se poate aplica tratamentul precum i predicia speranei de
supravieuire a celei sau pisicii respective.

CONCLUZII

1. n studiu au fost luate i examinate histologic un numr de 76 de tumori


maligne, din care 15 prezentau leziuni hiperplazice/displazice n esutul mamar
adiacent, cele 15 cazuri provenind de la 14 cele i de la o pisic.
2. Rasele de cini cu tumori mamare care prezentau leziuni
displazice/hiperplazice au fost: Caniche (6 cazuri), Cocker (3 cazuri), Ras
comun (2 cazuri), Vijla maghir, Pitt-Bull, Dog german (cte 1 caz).
3. Cele 15 cazuri au avut vrste cuprinse ntre 5-12 ani, cele mai multe
leziuni displazice fiind la vrsta de 10 ani (5 cazuri), media fiind de 8,3 ani.
4. Mamelele la nivelul crora s-au evideniat cele mai multe
hiperplazii/displazii au fost: M5 46,1%, M4 38,4%, M3 7,6% i M2 7,6%.

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5. Tipurile de neoplasme diagnosticate: carcinom tubular complex (5


cazuri), carcinom tubulo-papilar complex (2 cazuri), carcinom tubulo-papilar
simplu (2 cazuri), carcinom solid (1 caz), carcinom complex n tumor mixt
benign (3 cazuri), carcinom lobular in situ (1 caz), sarcom mamar (1 caz).
6. Leziunile displazice/hiperplazice evideniate au fost:
- hiperplazii lobulare: adenozis simplu (6 cazuri), epiteliosis tipic (12
cazuri), epiteliosis atipic (1 caz);
- hiperplazii ductale: epiteliosis tipic (4 cazuri), papilomatoz (1 caz);
- displazii chistice: 8 cazuri;
- ectazii ductale: 3 cazuri.
Pentru a efectua un examen histopatologic ct mai complet recomandm ca
prelevarea probelor s se fac i cu parenchimul mamar adiacent tumorii,
deoarece anumite leziuni hiperplazice cresc foarte mult riscul apariiei de tumori
mamare: adenozisul, hiperplaziile ductale epiteliale tipice, papilomatoza ductal,
de 2 ori, iar hiperplazia ductal i lobular atipic (ADH- ALH) de 4 - 5 ori.
Terapia chirurgical a tumorilor mamare trebuie s aib n vedere i excizia
acestor leziuni hiperplazice care sunt puncte de recidiv pentru tumorile mamare.

BIBLIOGRAFIE

1. Azzopardi J.G., 1979 - Papilloma and papillary carcinoma in problems in


breastpathology. Azzopardi JG ed. Philadelphia. Sauderrs 150-166.
2. Baba A.I., 2002 - Oncologie comparat, Editura Academia Romn, Bucureti
3. Catallioti L., Bianchi S., Ciatto S., 1999 - Patologia mammaria benigna in
Senologia oncologica. Veronesi U. Ed. Milano Masson, 194-197.
4. Gobbi H., Jensen R.A., Simpson J.F., Olson S.J., Page D.L., 2001 - Atypical
ductal hyperplasia and ductal carcinoma in situ of the breast associated with
perineural invasion, Hum Pathol, Aug. 32(8):785-90.
5. Misdorp W., Else R.W., Hellmen E., Lipscomb T.P., 1999 - Histological
classification of mamary tumors of the dog and cat.
6. Page D.L., Anderson T.G., 1987 - Papilloma and related lesions, in Page DL,
Anderson TG ed. Diagnostic Histopathology of the breast. New York, Churchill
Livingstone, 104-119.
7. Page D.L., Rogers L.W., 1992 - Combined histologic and cytological criteria for the
diagnosis of ductal atypical ductal hyperplasia. Hum Pathol 23:1095-1097.
8. Purcel C.A., Norris H., 1998 - Intraductal proliferation of the breast: a review of
histologic criteria for atypical intraductal hyperplasia and ductal carcinoma in situ,
including, apocrine and papillary lesions. Ann. Diagno. Pathol. 2(2): 135-145.
9. Shaaban A.M., Sloane J.P., West C.R., More F.R., Jarvis C., Williams E.M.,
Foster C.S., 2002 - Histopathologic types of benign breast lesions and the risc of
breast cancer, Am J Surg Pathol, Apr., 26(4): 421-30.
10. Shin S.J., Rosen P.P., 2000 - Pseudolactational hyperplasia: a primary diagnosis in
mamographically detected lesions of the breast and its relationships to cystic
hipersecretory hiperplazia, Am J Surg Pathol, Dec., 24(12): 1670-4.
11. Usha R., Vertes D., 1996 - Breast papillomas with atypical ductal hyperplasia; A
clinical pathologic study, Hum Pathol. 27(11): 1231-1228.

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FATTY LIVER IN NEWBORN AND YOUNG PIGLETS


B. SAVI1., H. AMANC2., V. STOJI2., V. IVETI1
1
Institute of Veterinary Medicine of Serbia, Belgrade
2
Faculty of Veterinary Medicine, Belgrade

Fatty liver occurs quite rare in newborn and young piglets. By


some authors it can be develop already during intrauterine live of piglets.
Pathogenesis of fatty liver metamorphosis in newborn piglets is still
unknown, presupposes that is consequent of lipotropic substances or
biologically importantly proteins deficit in sows feed in late gestation
period. Also the same changes could be effect of contamination of sows feed
with mycotoxins, which inhibit synthesis of appoproteins like one of the
important carrier of fats from liver or they contaminate sows milk. Authors
described clinical picture and pathomorphologycal changes on fatty
metamorphosed livers and discussed about ethological factors responsible
for fatty liver appearances in newborn and young piglets in first week of
live.

Key words: piglets, fatty liver, micotoxins

INTRODUCTION

Enzootic hepatosis, liver degeneration, hepatodistrophy and steatosis are


synonyms for fatty degenerations of the pig liver. It appears rarely on single or
many newborn or nursery piglets in the same litters, most often between 1 and 14
days of piglets live. According to Stamatovic and Samanc, 2001 this condition in
newborn piglets may already appear within intrauterine live apropos though last
third period pregnancy of sows. Herds with frequently appearance of this
condition have severe loss of piglets without clear expressed clinical symptoms.
Pathogenesis is still unknown, although central role within steatos
metamorphosis of liver have fats metabolisms disorders (Stojic, 2004). Disorders
can be qualified with many different factors, while in industrial pig productions
accent is applied on nutritive aspects (Peader and Brendan, 2001). Hypoxia
associated with lipotropic substances deficiencies during intrauterine live of
piglets apropos in late gestation pregnant female nutrition are triggers for
irreversible metabolic processes in hepatocytes appearance (Thomas at all, 1972;
Knezevic and Jovanovic, 1999). In ethiopathogenesis of liver degeneration
secondary metabolites of mould-mycotoxins participate significantly, which affect
in many different levels cells metabolism by inhibition of lipotropic essences
substances synthesis, which is essential in fatty transport from liver (Pegorier at
all., 1983). Also fatty liver increases the risk for infectious diseases and therefore
may be a causative factor in development of postnatal bacterial infections (Martini
at all., 2001).

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MATERIAL AND METHODS

We followed health status on three pigs farms and systematically noted all
relevant criteria in connection with health. First two, were small individual farms
with 40 sows capacity, third, were big industrial type farm with 1500 sows
capacity. We noted liver fatty metamorphosis in nursery piglets 1 to 10 days old.
Mortality rate increased on 15% on two small farms and 17% on third farm in first
10 days postnatal nurseries live. In addition, piglets were avital, born with less
than 1200 g body weight and though first days of suck period they were loosing
their weight and rapidly retarded. Some piglets were having non infective
diarrhea. In severe cases in some litters piglets were depressed, mostly lay on
creep and often were crashed by dams.
On third farm besides described clinical picture, hyperemia and vulvae
edema, skin necrosis on tail and carpal joints were noted. This symptoms are
related with mycotoxicosis respectfully zearalenontoxicosis. Unfortunately we did
dont have access to nutrient contents in sows feed ingredients on both smaller
farms. Only we were told that during all gestation period nutrition was not
modulated according to pregnant sows needs. On third farm manly component in
nutrient content was corn (around 60%) wheat grain (around 20%) and sunflower
meal (around 10%). High levels of mycotoxins, 9.2 mg/kg of zearalenone and 2.4
mg/kg of ochratoxins was detected in diet samples for pregnant sows.
Morphopathological exam was provided in all died piglets. Twenty tree
piglets originated from first two farm and twelve piglets from third farm were
autopsied.
Liver and kidneys tissues were fixed by immersion in 10% buffered
formalin. Fixed tissue was dehydrated, embedded in paraffin wax, sectioned at 5
m, and stained with haematoxylin and eosin (H&E). Cryosections tissues were
stained with Sudan III.

RESULTS

Gross pathology:
All examined carcasses were very emaciated. Macroscopically livers were
enlarged with rotund edges and intense capsules. In lightly cases (26 piglets, 75%
of total number) livers were decolorized with yellow - reddish foci, which were
markedly accented compared with normal tissue. At 9 piglets (25% of total
number) cases were severe with diffuse clay liver tissue colorization and pulpy
consistency. Cut surface bulge when incised. Liver parts moreover all lobes
floated when was placed in water in almost all cases.
Total or partial liver ruptures with sub capsular or intra abdominal blood
extravasations were detected in 7% cases .
Besides described morphopathological changes on liver macroscopically
kidneys were pale and yellowish, rare with cortical punctiform hemorrhage in all
died piglets originated from third farm.

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Microscopic finding:
Under light microscope perinuclear hepatocytes vacuolization were
detected. Usually one small vacuole was detected. In some parts of liver tissue
vacuoles coalesced to form large empty spaces with consequent nucleus
dislocating on cells periphery. Regressive changes were commonly located on
lobe periphery or they have panlobullary prevalence.
In kidneys epithelial cells, especially in proximal convoluted tubules
epithelias many small vacuoles whether diffuse filled cytoplasm was detected.
Vacuoles from cryostat sections liver tissue stained with Sudan III were
orange colorized and undoubtedly confirmed fat origin apropos liver fatty
metamorphosis. Parenchyma cells of the kidneys in cryosections contained many
orange fat droplets in cytoplasm as well.

DISCUSSION

To complete understand liver fatty metamorphosis mechanisms we will


briefly specified basic lipids metabolic pathways in organism and liver role in this
processes.
By tasks in organism lipids are divides on labile and stabile
components. Labile lipids originate from feed, or generates from carbon hydrates
and some amino acids. They consist of neutral lipids, which are used for normal
substance exchange in hepatocytes, at why excesses are accumulated in fat depots
in organisms. From fat depots neutral lipids could be activated and transported to
the liver or other organs.
In lipids metabolisms liver has several roles: fatty acids -oxidation to
acetyl coenzyme-A, which is included in Krebs cycles of tricarbon acids from
whose acquire energy; free fatty acids (FFA) esterification to triglycerides which
is transported in blood plasma like very low density lipoproteins (VLDL) banded
with apoproteins; phospholipids and cholesterol synthesis from fatty acids.
From classical pathologic view, four factors are important for fatty changes
appearance in hepatocytes: increase lipids intake after intestinal resorbtion;
increase fatty acids intake from fat depots; decrease -oxidation process in
hepatocites; decrease synthesis of apoproteines and thereby decrease productions
and release of lipoproteins (VLDL) from hepatocytes.
Causes of fatty metamorphosis in hepatocytes are numerous but most often
are: hypoxia with very low oxygens levels in blood; anemia; CO poising;
oxygens free radicals poising; mycotoxins intoxications with consequences
nonsaturated fat acids hepatocytes membranes phospholipids peroxidations;
essentials fatty acids, methionin, holing, B- complex vitamins especially B12 lack.
All mentioned mechanisms dont have equal significance in hepatocytes
fatty changes. Not only one, but also two or more mechanisms could be involved

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in disorders pathogenesis. Last tree are especially accentuated in connections with


diet hepatosis in pigs.
As mentioned before phospholipids have central role in lipotrop and
antilipotrop events in liver. Phospholipids molecules are composed from one
hydrophilic and one hydrophobic end. Polarized choline group in ester
conjunction with glycine and phosphoric acid make hydrophilic end, and
nonpolarized fatty acids residues make hydrophobic end. Substances with hereby
physically and chemically attributes react as emulgators. Consequently
phospholipids significantly affect on hylomicrons - triglycerides forming which
important transport form of neutral fats from liver.
On the other hand, cholines participate in phospholipids synthesis in cells
like methyl group donor in reactions of transamination. In hepatocytes it has an
important role in lipoids, especially biological membranes phospholipids and
acetylcholine synthesis. Choline is quatern ammonium basis esterifically banded
with glycine and phosphoric acid embedded in phospholipids lecithin. It is
synthentised from serine amino acids products (ethanolamine) by decarboxilation.
Ethanolamine (cholamine) will be methylated by methionine CH3 labile groups. In
this reaction, as methyl and transmethyl cofactors, thetrahydropholic acid and
cobalamin B12 vitamin participate too. The coenzyme of vitamin B12 is a carrier of
methyl groups. Because of its methyl transfer role, vitamin B12 is active in the
synthesis of the amino acid methionine from its precursors, homocysteine. The
coenzyme transfers methyl groups from methyl folate, a derivate of folic acid, to
homocysteine and methionine is formed. Second role of vitamin B12 is
izomerisation of malonyle coenzyme A to succinyle coenzyme A.
Therefore choline and its precursors (serine, ethanolamine and methionine)
deficit are relate to irreversible processes in hepatocytes, which cause fatty liver
metamorphosis. Choline deficiency decreases phospholipids syntheses, which
assist in fats transport from liver. Vitamin B12 plays a secondary role in the lipid
pathway.
Moreover very complexly micotoxins reaction mechanisms inhibited
synthesis of phospholipids like overall effect.
A complexes mechanism which is involving in choline deficit and its
influence on phospholipids synthesis is schematized below.
Altogether suspend of phospholipids synthesis by choline deficit or either
another reason, disarrange fats transport between liver and fat tissue which itself
repercuting on fats accumulation in the liver.
Fatty liver in nursery piglets is most likely consequence of pregnant sows
nutritive deficits respectfully essential nutritive substances principally lipotropic
factors choline, methionine, bethaine and vitamin B12 lack.
By Forenbacher, 1993 poor choline sows nutrition during gestation
qualified avital newborn piglets with fatty liver. Besides induced, trophopathic
liver damage in fetuses is result of high valuable proteins deficits. Undoubtedly

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those mycotoxins play important role in this changes most likely inhibiting
appoproteines synthesis, which is required in fats transport from liver by
lipoproteins complexes.

CONCLUSIONS

Fatty metamorphosis in nursery piglets is consequence of:


- Pregnant sows nutrition deficits (choline, methionine, bethaine,
vitemine B12 and biologically highfalutin proteins deficits), which
probable occurred on two small farms;
- Sows feed contamination with mycotoxins, which is inhibiting
appoproteines synthesis, which were detected in sows feed samples;
- Fatty liver conditions in newborn piglets could increase mortality rate
in first 5 to 6 day nurseries live;
- Fatty liver changes could be detected in stillborn piglets, what is in
agreement with thesis that changes appeared in last third of gestation.

REFERENCES

1. Stamatovi S.M., H.A. amanc, 2001 - Swine diseases. VKS. Belgrade


2. Stoji V., 2004 - Veterinry phisiology. Scientific book KMD. Belgrade
3. Peader G. Lawlor, P. Mrendan Lynch, 2001 - Mycotoxins in pig feed 2: clinical
aspects. Irish Veterinary Journal, volume 54 (4).

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4. Bownhemer T., Richard J. Havel, John A. Long, 1972 - Physiological fatty liver
hyperlipemia in the fetal guinea pig: chemical and ultrastructural characterisation.
Journal of Lipid Research, Vol. 13, 371 382.
5. Kneevi Milijana, M. Jovanovi, 1999 - General pathology. Makrije, Belgrade.
6. Pegorier J.P., P.H. Duee, J. Girard, J. Peret, 1983 - Metabolic fate of non-esterified
fatty acids in isolated hepatocytes from newborn and young pigs. Biochemical J.
212, 93 97.
7. Martini, Wenjun Z, Irtun, Oivind, Chinkes, David L, Rasmussen, Blake, 2001 -
Alteration of hepatic fatty acid metabolisms after burn injury in pigs. Journal of
Pareneteral and Enternal Nutrition, Nov/Dec 2001.
8. Forenbacher S., 1993 - Digestive system and metabolisms clinical pathology. NAZU,
kolska knjiga Zagreb.

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MODIFICRI MORFOLOGICE ALE ORGANELOR


GENITALE N PSEUDOHERMAFRODITISMUL MASCULIN
LA CAPRA DOMESTIC (CAPRA HIRCUS)
T.M. TOPAL, G. BALMU,
C. COTEA, S. PACA, O.Z. OPREAN
U.S.A.M.V. IAI

At a 6 months old goat with pronounced female phenotipical aspect,


but with a very developed clitoris was made euthanasia.
Necropsy exam shows two testicles inside the pelvis and genital ways
with tubular aspect.
Histological exams reveal the testicular parenchyma with seminal
epitheliums vacuolized and with rare spermatogonies and interstitial Leydig
glands rudimentary.
The tubular organ is uterus, with lamina propria intensively
collagenized and underdeveloped endometrial glands.
In the wall of uterus are incorporated glandular structures with
typical aspect for the seminal vesicles.

Key words: Capra hircus, hermaphroditism, malehermaphroditism

Hermafroditismul poate fi considerat ca fiind adevrat de tip bilateral,


unilateral i alternant. Hermafrodismul fals sau pseudohermafroditismul poate fi
de tip masculin i feminin.
Hermafroditismul fals (pseudohermafroditismul) masculin, const n
prezena la acelai individ a gonadelor masculine i a cilor genitale feminine.

MATERIAL I METOD

Cercetrile noastre au avut ca obiect de studiu o capr din rasa Carpatin


n vrst de 6 luni. Din punct de vedere al aspectului fenotipic acesta era femel,
deoarece glanda mamar era prezent.
Metodele de lucru folosite au fost eutanasia cu soluie T61, examenul
necropsic i examenul histopatologic.
Pentru realizarea examenului histopatologic s-au extras fragmente tisulare
de 1-2 cm din aparatul genital care au fost fixate n formaldehid 10% soluie
apoas, inclus n parafin , secionate la 5m i colorate cu metoda
Hematoxilina-Eozina-Albastru de metil (HEA).

REZULTATE I DISCUII

Dup efectuarea investigaiilor morfologice macro- i microscopice a fost


pus diagnosticul de pseudohermafroditism masculin deoarece aparatul genital

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(Fig. 1) prezenta gonadele de tip masculin (Fig. 2, 3) aceasta n condiiile n care


la exterior capra prezenta aspect fenotipic femel prin prezena glandei mamare i a
vulvei (Fig. 4).

Fig. 1 - Aparatul genital Fig. 2 - Gonada masculina dreapta

Fig. 3 - Gonada masculina stng Fig. 4 - Clitoris

n urma examenului histologic, la nivelul testiculelor se observ albugineea


intens colagenizat i vascularizat (Fig. 5) i vacuolizarea epiteliului tubilor
seminiferi (Fig. 6). La nivelul epiteliului seminifer se observ o distrofie hidric
(Fig. 7), iar epididimul prezint o hipoplazie epitelial i fibroz interstiial
(Fig. 8).

Fig. 5 Albuginee colagenizat i Fig. 6 Vacuolizarea tubilor seminiferi


vascularizat Col. HEA, x100 Col.HEA, x400

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Fig. 7 Epiteliul seminifer.Distrofie hidric Fig. 8 Epididim. Hipoplazie epitelial


Col. HEA, x400 Col. HEA, x100

n urma examenului histologic la nivelul uterului se constat hiperplazia


musculoasei (Fig. 9), hiperplazia limfohistiocitar i colagenizare n lamina
propria (Fig. 10, Fig. 11). Totodat se observ chistizarea glandelor endometriale
(Fig. 12), precum i hiperplazie i cheratinizare epitelial la nivelul mucoasei (Fig.
13), i vezicula seminal intraparietal (Fig. 14).

Fig. 9 Uter. Hiperplazia musculoasei Fig. 10 Uter. Hiperplazie limfohistocitar n


Col.HEA, x100 lamina propria. Col. HEA, x400

Fig. 11 Uter. Colagenizare n lamina propria Fig. 12 Uter.Chistizarea glandelor


Col. HEA, x400 endometriale. Col.HEA, x400

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Fig. 13 Uter. Mucoasa. Hiperplazie si Fig. 14 Uter. Vezicula seminala


cheratinizare epiteliala. Col.HEA, x400 intraparietala. Col.HEA, x100

CONCLUZII

Hermafroditismul fals (pseudohermafroditismul) masculin const n


prezena la acelai individ a gonadelor masculine i a cilor genitale feminine.
La o capr n vrsta de 6 luni cu aspecte fenotipice femele ( glanda mamar
dezvoltat anatomic normal ) dar cu un clitoris dezvoltat, examenele radiologic i
ecografic evideniaz la intrarea n cavitatea pelvin dou formaiuni ovoide cu
diametrul de 1,8 cm foarte asemntoare cu testiculele.
Examenul necropsic evideniaz pe ambele pri testicule i epididim, un
conduct genital cu diametrul de 0,5 cm i un clitoris foarte dezvoltat .
Examenele histologice relev structuri testiculare epididimare afuncionale:
albuginee intens colagenizat, distrofie hidric a epiteliului seminifer, glande
interstiiale Leydig rudimentare, hipoplazie tubular i fibroz interstiiala n
epididim.
Organul tubular are structuri tipic uterine dar modificate spre fenomene
intens proliferative: miometru hiperplaziat, proliferri polimorfe n lamina
propria, glande endometriale chistizate, hipoplazia i cheratinizarea endometrului.
n peretele uterin se evideniaz dou formaiuni bine delimitate cu aspecte
structurale caracteristice veziculelor seminale.

BIBLIOGRAFIE

1. Cernescu H., 1989 Patologia reproduciei i clinic obstretical. Lito.


USAMVB.,Timioara;
2. Cotea V.C., 2003 - Histologie special, Ed. Tehnopress Iai;
3. Groza I., Munteanu M., 2002 - Elemente de fiziologia reproduciei la animale. Ed.
Academic press, Cluj-Napoca;
4. Paul I., 1990 - Morfopatologia aparatelor i sistemelor organice. Soc. de Med. Vet din
Romania, Buletin Inf. nr. 23-24., , Bucureti;
5. Runceanu L., Cotea C., 2001 Reproducie, obstetric i ginecolgie veterinar. Ed.
Ion Ionescu de la Brad, Iai;

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FIBROADENOMUL MAMAR LA PISIC


MAMMARY FIBROADENOMA IN CATS

Gabriela URSACHI1, Otilia COOFAN2,


Iulia FLORITEAN1
1
D.S.V.S.A. L.V.S. Iai
2
F.M.V. Iai

Fibroadenoma, also known as the benign or fibroglandular


mammary hypertrophy, feline mammary adenomatosis, mammary
fibroadenomatous hyperplasia was diagnosed in a 4 years old cat. Clinical
investigation revealed small nodules, about 5 mm in diameter in the right
caudal pectoral mammary gland. The nodules, well circumscribed, were
rounded, relative smooth, mobile, with a gummy consistence section
surface having white, shiny and fasciculate pattern.
Microscopically, the tumor parenchyma consists of elongated ,
irregular and widely branched tubular systems covered by mono- or
multistratified columnar epithelium without notable atypia or mitotic
activity.
The proliferating stroma forms massive and concentric cuffs of fine
fibers consisting in fine fibers and spindle cells close to the tubes and thick
collagen bands farther . The peripheral demarcation of the lesion is
produced by the condensation of the mammary stroma. The aspects of the
lesion correspond to the intermediary stage between the peri- and
intracanalicular types of fibroadenomas.

Key words: fibroadenoma, hypertrophy, adenomatosis.

Fibroadenomul mamar este cunoscut i sub denumirea de hipertrofie


mamar benign sau fibroglandular, hiperplazie fibroadenomatoas
mamar i adenomatoza mamar felin
Este descris i la cine i semnalat foarte rar la capr i bivoli. La femei
ocup locul al treilea n mastopatii dup mastoza Reclus i carcinoame; la brbai
se dezvolt dup tratamentul intens cu estrogeni n cancerul de prostat.
Afecteaz cel mai frecvent pisicile tinere n plin activitate genital, n faza
de estru, n gestaie, dup tratamentele cu medroxyprogesteron-acetat o lung
perioad de timp pentru prevenirea estrului, precum i la motani dup
administrarea compuilor progesteronici. Afecteaz una sau mai multe glande,
uni- sau bilateral i se manifest prin noduli solitari sau multipli (n 20% din
cazuri), cu diametrul variind ntre 1-2 mm i maximum 3 cm la pisic i 5-7 cm
la cea. Nodulii sunt rotunjii, sferoizi sau polilobulai, fermi, elastici, bine
delimitai, dar nencapsulai, mobili fa de esuturile vecine, enucleabili; culoarea

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este alb-sidefie, alb-rozat sau brun roiatic, suprafaa de seciune fiind umed i
strlucitoare i proeminent fa de esutul mamar adiacent (Sternberg, 1989).
Se comport ntotdeauna benign, evolueaz lent, uneori regreseaz spontan
sau dup ovariectomie.

MATERIAL I METOD

Materialul de studiu a fost reprezentat de o pisic n vrst de 4 ani, care


prezenta noduli mici, n glanda mamar pectoral caudal dreapt. Nodulii
tumorali au fost prelevai n totalitate, fixai n formaldehid 10%. Fragmente de
noduli au fost incluse n parafin, secionate la 5 micrometri i colorate prin
metodele HEA, PAS i impregnare argentic Gmori.

REZULTATE I DISCUII

Macroscopic, la examenul clinic s-au sesizat noduli mici cu diametrul de


5 mm n glanda mamar pectoral caudal dreapt.Nodulii erau bine delimitai,
netezi, mobili, de consisten gumoas, cu suprafaa de seciune alb-sidefie i cu
aspect fasciculat.
Microscopic, parenchimul tumoral este mixt, format din structuri epiteliale
i esut conjunctivo-vascular. Cazul luat n studiu exprim un amestec din
subtipurile de fibroadenom pericanalicular i intracanalicular.
La periferia nodulului tumoral se identific capsula groas i dens
format prin condensarea esutului conjunctiv fibros mamar, mpins de creterea
tumorii (fig. 1). Din capsul se desprind septe conjunctivo-vasculare compacte
care alctuiesc stroma i delimiteaz lobulii tumorali (fig. 2) constituii din
parenchimul tumoral mixt (epitelial i conjunctival) (fig. 4).

Fig.1. Fibroadenom mamar la pisic. Fig. 2. Fibroadenom mamar la pisic. Septe


Capsula, sept interlobular. PAS x40 interlobulare. HEA x 100

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Fig. 4. Fibroadenom mixt. Sisteme tubulare


Fig. 3. Fibroadenom. Gomori x 100 alungite i amplu ramificate i zone
fibromixomatoase. HEAx100

n tipul pericanalicular, componenta epitelial este organizat n tubuli mici


i rotunjii sau ovalari, cptuii cu ambele tipuri de celule (luminale i
mioepiteliale), fr atipii sau cu lumene deschise i uneori cu secreie bazofil i
PAS pozitiv.

Fig. 5 Fibroadenom intracanalicular mamar Fig. 6. Fibroadenom intracanalicular la


la pisic. Secreie i tubi nconjurai de pisic. Componenta epitelial cu ambele
esut fibros edemaiat. PAS x 100 tipuri de celule. HEA x200

Componenta fibromatoas constituit din fibre fine i celule fibroblastice


fusiforme, cu un numr variabil de celule adipoase, se nfoar n jurul tubulilor
formnd manoane concentrice.
n subtipul intracanalicular, tubulii epiteliali se alungesc i se ramific, se
turtesc i se deformeaz datorit mugurilor conjunctivi care mping pereii i
proemin n lumen. Cu timpul ncep s semene cu coarnele de cerb. Celulele
epiteliale au tendin spre pleomorfism. Componenta conjunctiv a
fibroadenomului este texturat lax, mixomatoas, fiind format din celule

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fusiforme i stelate, derivate din celulele mioepiteliale spaiate de substan


intercelular mucoid abundent (fig. 5, 6).
Proliferarea stromei conjunctive este mult mai difuz i aspectul lobular
mult mai ters.

CONCLUZII

Se prezint aspectele macroscopice i microscopice ale fibroadenomului


mamar , diagnosticat la o pisic de 4 ani.
Microscopic,aspectul corespunde stadiului intermediar de evoluie ntre
tipurile pericanalicular i intracanalicular vegetant.

BIBLIOGRAFIE

1. Baba A.I., 2002 Oncologie comparat - Editura Academiei Romne, Bucureti


2. Bostock D.E., 1986 Canine and feline mammary neoplasms Br Vet J.
142(6):506-515,Nov-Dec;
3. Hayden D.W.Y., Johnston S.D., Kiang D,T., Johnson K.H., Barnes D.M., 1981
Feline mammary hypertrophy/fibroadenoma complex: clinical and hormonal
aspects, Am J Vet Res.; 42(10):1699-1703, oct;
4. Jubb K.V.F., Kennedy P.C., Palmer N., 1993 Pathology of Domestic Animals,
Fourth Edition, V.M. 3 Academic Press, New York;
5. Sternberg S.S., 1989 - Diagnostic surgical Pathology, Raven Press, LTD, New York

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MASTOPATIA FIBROCHISTIC LA CANIDE


FIBROCYSTIC MASTOPATY IN DOGS

Gabriela URSACHI
D.S.V.S.A.-L.V.S. Iai
Otilia COOFAN
U.S.A.M.V. Iai
G. HUEAL
D.S.V.S.A.-L.V.S. Buzu
I. PAVLOVIC
Scientific veterinary Institute of Serbia

The fibrocystic or sclerocystic mastopathy, cystic mastosis, Reclus


disease, Schimmellbusch disease mamary fibroadenomatous hyperplasia is
the most common benign disease, well known in the women, described also
in bitches and rarely in cats.
In our investigations, the disease was localized in the caudal
mammary glands III-V in three bitches aged 6, 7 and 9, had 5-18 cm in long
diameters, and irregular shapes determined by the conglomeration of
small, hard, withe-yellowish nodules combined with fluctuant cysts of
various sizes, with an yellow or brown content, all sustained by a sclerotic
stroma. Gelatinous areas alterning with greasy foci and rough sandy
nodules were observed on the cut surfaces. Zonal lymphonodes were
slightly enlarged and had a firm consistency.
Microscopically, lesions are developed in TDLU. The cysts are
covered by a columnar, atrophic, papillary or partially exfoliated
epithelium and contain a PAS positive secretion with lipids, xanthomatous
cells, cholesterol, calcium precipitates, dead cells or cellular dust. These
aspects are associated with ductular or alveolar epithelial hyperplasia,
proliferation of the connective tissue, hyalinization of the collagen,
multiplication of the myoepithelial cells forming a myxoid or chondroid
stromal matter, stromal infiltrations with mononuclear cells and
hemosiderophages and sometimes a pericystic chronic, granulomatous
reaction with giant cells. A lymphoplasmocytic hyperplasia and sinusal
reactive histiocytosis in the zonal lymphonodes were also noticed. Some
zones with massive epithelial proliferation, high mitotic ratio, moderate
pleomorphism , and discontinous basal membranes suggest the potenial for
malignant transformation of this disease.

Key words: sclerocystic mastopathy, dogs

Mastopatia sclerochistic (fibrochistic) sau boala Reclus este cea mai


banal afeciune mamar la femeie, constatat la 53% din cazurile necropsiate cu
frecvena cea mai mare n a patra decad a vieii (Niculescu, 1980). Procesul

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patologic afecteaz una sau ambele mamele, debuteaz i evolueaz n TDLU i


se caracterizeaz prin hiperplazii epitelio-mezenchimale i dilataia chistic a
unitilor asociate cu scleroz i metaplazii ale stromei.
n oncologia veterinar, mastopatia este semnalat la carnivore, la canidele
tinere fiind mai frecvent dect la pisicile de orice vrst. Descrierea tabloului
lezional este lapidar i vag, cu att mai mult cu ct afeciunea denumit mastoz
sau mastopatie sclero(fibro)chistic sau chiar hiperplazie fibroadenomatoas
mamar este asimilat categoriei de fibroadenom.

MATERIAL I METODE

Materialul de studiu a fost reprezentat de 3 canide femele care prezentau


procese tumorale ale glandelor mamare. De la fiecare caz s-au prelevat
fragmente de esut tumoral sau chiar tumora n totalitate.
Fragmentele recoltate au fost fixate n formaldehid 10 % sau lichid fixator
Bouin, incluse n parafin, secionate la 5 micrometri i colorate prin metodele
HEA, PAS, Pappenheim, Van Gieson.

REZULTATE I DISCUII

Aspecte macroscopice.
Cazul 1. Femel de ras comun n vrst de 9 ani a prezentat dou tumori
localizate n lanul stng. Glanda mamar M3 stng avea diametrul de 18 cm i
un contur neregulat datorat multitudinii de chisturi mici, izolate sau conglomerate,
i prezenei chisturilor mari alternnd cu structuri nodulare mici, glbui i dure
care dau la palpare senzaia de alice i bile de plumb. Chisturile mari sunt bine
circumscrise, relativ mobile fa de esuturile adiacente, elastice i au un coninut
galben-verzui sau brun-roiatic sub tensiune.

Fig. 1. Mastopatia chistic.


Aspectul exterior al tumorii.
M3 stng

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Glanda mamar M5 stng avea diametrul de 5 cm i se prezenta ca o mas


tisular bine delimitat , uor indurat avnd n suprafaa de seciune zone
gelatinoase i noduli albi-glbui i crnoi. Limfonodurile inguinale superficiale
homolaterale apreau mrite, avnd lungimea de 3 cm i grosimea de 1 cm. Un
limfonod inguinal profund era de asemeni uor mrit i avea o consisten ferm.
Cazul 2. Femel de ras comun n vrst de 7 ani, a prezentat o tumoare
localizat n M4 din lanul drept , cu dimensiunile de 12 cm lungime, 8, 5 lime i
10 cm nlime. Aspectul exterior era comparabil cu un ciorchine de strugure, iar
suprafaa de seciune cu un vaier datorit cavitilor chistice pline cu o substan
gelatinoas galben sau roiatic.
Cazul 3. Femel de ras comun n vrst de 6 ani, a prezentat o formaiune
nodular cu diametrul de 5 cm localizat n M5 lanul drept, relativ bine
circumscris, avnd zone albicioase i dure i structuri cavitare cu coninut glbui.

Aspecte microscopice.
Mastopatia debuteaz i evolueaz n unitile lobulare alveole-ducte
terminale (engl. terminal duct lobular unit sau TDLU) i se caracterizeaz prin
hiperplazia de tip adenotic i dilataia chistic a unitilor, asociate cu dezvoltarea
esutului fibros, colagenizare, hialinizare i metaplazie stromal. Coninutul
chisturilor este variat fiind reprezentat de material proteic amorf sau din secreie
glicoprotidic intens PAS pozitiv (fig. 3) n care se pot gsi celule epiteliale
descuamate, lipofage cu citoplasma spumoas, picturi lipidice i grsimi
saponificate, detritus celular, depozite de calciu, cristale negative romboidale de
colesterol libere sau compactate sub form de tablete, rezultate din stagnarea
secreiei i dezintegrarea lipofagelor.
Dilataia saccular alveolar i ductular genereaz chisturile mici, multiple
i adiacente, delimitate de esut conjunctiv dens care, prin ruperea pereilor
despritori, conflueaz n chisturi mai mari (fig. 2). Acestea sunt cptuite cu
celule epiteliale secretorii (luminale) monostratificate, prismatice, cubice sau
atrofiate prin compresiune, dublate de celule mioepiteliale, formnd un tapet
neted n forma nepapilar. n forma papilar, are loc hiperplazia epitelial cu
pluristratificare (fig. 4) urmat de formarea unor proeminene intracavitare
constituite din axe conjunctivo-vasculare ramificate, arborescente, acoperite cu
epiteliu bimorfic ale crui celule luminale pot fi bogate n glicogen. n unele
chisturi voluminoase hiperplazia epitelial poate fi parial sau complet disprut
datorit descuamrii celulelor n cavitate.Se remarc prevalena net a celulelor
mioepiteliale n procesul proliferativ; plasate ntre celulele epiteliale secretorii i
membrana bazal, frecvent vacuolizate, prin multiplicare excesiv pot genera
noduli adenomioepiteliomatoi stromali sau intrachistici.
n interstiiul tumorii, la periferia chisturilor se gsesc numeroase
mononucleare (monocite-macrofage, limfocite, plasmocite), frecvente
hemosiderofage, dar i celule gigante multinucleate care sugereaz deversarea

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coninutului chisturilor n strom i iniierea unor reacii inflamatorii de tipul


granulomului de corp strin.

Fig. 2. Mastopatia fibrochistic. Fig.3. Conglomerat de chisturi cu secreie.


Conglomerat de chisturi. HEA x 40 PAS x 40

Fig. 4. Hiperplazie epitelial intrachistic. Fig. 5. Metaplazie cartilaginoas stromal.


PAS x 40 Pappenheim x 100

n unele cmpuri (cazul 1), stroma tumorii conine zone de metaplazie


cartilaginoas, bazofil n HEA, PAS pozitiv i metacromatic la MGG i
Pappenheim (fig. 5).

CONCLUZII

1. Mastopatia sclerochistic a fost identificat la trei femele de ras comun


n vrst de 6-9 ani, leziunile de dimensiuni mari (ntre 5 i 18 cm) fiind localizate
n mamelele caudale, multiple ntr-un singur caz i caracterizate prin consistena
chisturilor i a unor noduli mici, glbui, duri.

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2. Microcopic, mastopatia se dezvolt n TDLU i se particularizeaz prin


hiperplazia de tip adenotic a epiteliului bimorfic i dilataia chistic a unitilor
lobulare, asociate cu colagenizare, hialinizare i metaplazie condroid stromal.
3. Hiperplazia epitelial marcat, topirea membranelor bazale i tendina de
invazie a stromei, creterea raportului N/C i a indicelui mitotic, anizocarioza
pledeaz pentru potenialul de transformare carcinomatoas a mastopatiei.

BIBLIOGRAFIE

1. Manolescu N., Bolte S., Miclu I., 1993 - Oncologie veterinar, vol. II, Editura
Ceres, Bucureti
2. Misdorp W., 2002 - Tumors of the Mammary Gland. In. Tumors in Domestic
Animals. 4 th Ed.,Iowa State Press
3. Niculescu t., 1980 - Morfopatologia glandei mamare. n Moraru, I. (sub redacia)
Anatomie patologic, Editura Medical, Bucureti
4. Tavasoli F.A., Devilee P., 2003 - World Health Organization Classification of
Tumours. Pathology and Genetics of Tumours of the Breast and Female Genital
Organs. IARC Press: Lyon
5. Theilen H. Gordon, Madewell R. Bruce, 1978 Veterinary cancer Medecine,
Second Edition, Lea & Febiger, Philadelphia,

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ASPECTE HISTOPATOLOGICE N
TUMORILE SPONTANE LA CINE
HISTOPATHOLOGICAL ASPECTS IN SPONTAN TUMOURS IN DOGS

M. COMAN, I. OLARIU-JURCA,
A. STANCU, D. ROOGA
U.S.A.M.V.B. Timioara

We investigated histologically 54 tumours in dogs, taken from


animals within Timioara. Our researches aimed at the incidence of
spontan neoplasms, their major locations depending on breed, sex, age, and
also their histopathological forms.
Benign epithelial tumours (adenoma, papilloma) seem to be more
numerous (54.54%), while the benign mesenchymal tumours (fibroms,
leiomyoma) are slightly reduced (45.45%).
Regarding malign tumours, the proportion between epithelial
tumours (carcinoma, adenocarcinoma) and mesenchymal tumours
(sarcoma) is categorically in the favour of the epithelial ones (81.25%).
The degree of malignity of the tumours diagnosed histologically does
no always corelate with the macroscopical aspects, this aspect being more
evident in the case of adenocarcinoma.

Key words: dog, tumours, organs, histopathological forms.

Procesul neoplazic se caracterizeaz prin elemente constant comune


majoritii tumorilor, cum ar fi comportamentul biologic, anatomie, proliferare
necontrolat, invazivitate i metastazare, formarea de clone celulare noi i
ireversibilitate. O alt serie de proprieti comune sunt cele morfologice, ncepnd
cu aspectele moleculare i ultrastructurale, pn la aspectul histopatologic i
macroscopic al esutului neoplazic (5, 6, 7, 8).
Investigaiile ntreprinse au avut drept scop urmrirea incidenei
neoplasmelor spontane pe raza municipiului Timioara, a principaleleor localizri
n funcie de ras, sex sau vrst, precum i formele histopatologice ale acestora.

MATERIALE I METODE

Investigaiile prezentei lucrri au fost efectuate pe 54 tumori din diverse


esuturi i organe la cine de pe raza Municipiului Timioara.
Pentru examen histopatologic fragmentele prelevate din tumori au fost
fixate n formaldehid soluie 10 %, a urmat tehnica la parafin, secionate la 6
micrometri i au fost colorate prin metoda tricromic Masson modificat de V.
Ciurea cu albastru de metil pentru orientarea general, respectiv metoda Giemsa
pentru detalii celulare.

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REZULTATE I DISCUII

Rezultatele obinute n urma citirii preparatelor histopatologice sunt


consemnate n tabelele 1 i 2 privind diagnosticul tumorilor benigne i maligne la
cine.
Tabelul 1
Tipul i localizarea tumorilor benigne la cine

Nr. Denumirea Nr. Sex Vrsta


Nr. cazuri Localizarea
crt. tumorii cazuri M F ani
Mamel 5 - 5 10-12
1 Adenom 9 Pulmon 3 2 1 8-10
Prostat 1 1 - 10-12
2 Papilom 3 Cutanat 3 2 1 8-10
Uter 1 - 1 10-12
3 Fibrom 6 Vagin 1 - 1 8-10
Subcutanat 4 4 - 6-8
Uter 2 - 2 10-12
4 Leiomiom 4
Vagin 2 - 2 8-10
TOTAL 22 22 9 13

Din Tabelul 1 rezult c cele 22 tumori benigne se ncadreaz n grupa


tumorilor epiteliale (54,54%): adenom (9 cazuri), papilom (3 cazuri), respectiv
diferena (45,45%) aparine tumorilor mezenchimale: fibrom (6 cazuri), leiomiom
(4 cazuri).
Se poate deduce faptul c n cazul tumorilor benigne se nregistreaz o
evoluie relativ asemntoare la cele dou sexe, existnd o inciden maxim la
aceeai categorie de vrst (8-12 ani). Cu alte cuvinte, frecvena apariiei
tumorilor benigne nu este influenat de sex ci, n principal, de vrst.
Adenomul este tumora benign a epiteliilor glandulare; dup organizarea
celulelor hiperplaziate am identificat mai multe tipuri de adenoame: compacte,
tubulare, chistice i papilifere. A fost diagnosticat n 9 cazuri: mamela (5 cazuri);
pulmonar (3 caz) i prostat (1 caz).
Papilomul, tumora benign epiteliilor pavimentoase stratificate de tip
cornos, evolueaz prin trei tipuri de papiloame: hipercheratozice, acantozice i
fibropapilomul. A fost diagnosticat n trei cazuri la nivelul pielii, papilomul
acantozic.
Fibromul, tumora benign a esutului conjunctiv, a fost diagnosticat de noi
n structurile genitale la cele. n ambele localizri tumorile au fost alctuite din
fibre conjunctive organizate sub form de fascicule orientate n toate direciile.
Leiomiomul, tumora benign a esutului muscular neted, s-a diagnosticat n
uter i vagin. Este constituit din fibre musculare netede inegale dispuse n
fascicule sau cu orientri diferite.

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n Tabelul 2 sunt expuse tipul i localizarea tumorilor maligne la cine.


Aprecierea malignitii unei tumori, n funcie de origine (ncadrare
histopatologic) se face dup mai multe aspecte legate de morfologia celulelor
(aspectul nucleului, uniformitatea celular, prezena sau absena membranelor
bazale, tipul de cretere, frecvena diviziunilor i atipiilor de diviziune, raportul
nucleocitoplasmatic etc.).
Tabelul 2
Tipul i localizarea tumorilor maligne la cine

Nr. Nr. Sex Vrsta Metas-


Denumirea tumorii Localizarea
crt. Cazuri M F ani taze
Mamel 16 - 16 7-12
1 Adenocarcinom 2
Prostat 1 1 - 10-11
Mamel
2 Carcinom solid 2 - 2 10-12 2
3 Carcinom folicular Tiroid 1 - 1 9-10 -
4 Carcinom scuamos Cutanat 3 2 1 6-8 -
Mamel 1 -
5 Sarcom 1 4-6 -
Subcutanat 3 3
6 Seminom Testicul 2 2 - 8-10 -
7 Fibrosarcom esut conj. 3 3 - 6-7 -
TOTAL 32 11 21

Din tabelul de mai sus rezult c 59,37% din tumorile maligne au localizare
mamar, n timp ce diferena (40,62%) au localizri diferite (tiroid, testicul, esut
conjunctiv, subcutanat, etc.).
Frecvena tumorilor maligne n cazul femelelor crete odat cu vrsta, pn
n jurul vrstei de 12 ani, urmnd apoi o descretere mai mult sau mau puin lent.
n cazul tumorilor maligne la masculi, incidena este sczut pn n pragul
vrstei de 4 ani, dup care aceasta crete, i se menine la valori apropiate.
Adenocarcinoamele, tumori maligne ale epiteliilor glandulare, difer prin
aranjamentul celular i raportul dintre strom i parenchim. Adenocarcinoamele
identificate de noi se ncadreaz n tipurile tubular i papilar. Metastazele ntlnite
n dou cazuri au afectat limfonodurile lomboaortice i inghinale.
Carcinomul bronhoalveolar are o structur alveolar cu aspect glandular.
Histoarhitectonica pstreaz linii celulare tumorale att din alveole ct i din
bronhiile terminale.
Adenocarcinomul folicular se identific microscopic uor datorit
dispunerii celulelor n foliculi. Celulele tumorale sunt relativ mari, formnd
foliculi de diferite dimensiuni, forme i coninut coloid.
Leiomiosarcomul, tumora malign a fibrelor musculare netede, se
caracterizeaz prin fascicule celulare ai cror nuclei apar mult alungii, au indicele
mitotic ridicat i uneori cu microfocare de necroz.

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Carcinomul cu celule scuamoase este un neoplasm cu nalt grad de


malignitate, frecvent asociat cu dermatoza solar, fiind tumora celulelor stratului
malpighian din epiderm. Se recunoate dup prezena perlelor cheratozice, n
centrul crora se observ cheratina lamelar pe cale de necroz.
Fibrosarcomul este tumora malign a fibroblastelor, dar cu participarea i a
altor celule mezenchimale, respectiv a colagenului. Se constat o structur
format din fibroblaste cu aspect fusiform sau/i stelat, separate prin fascicule de
colagen puin abundent.
Sarcomul rotundocelular se recunoate dup parenchimul tumoral alctuit
din celule rotunde mici, cu nucleu mare, bogat n cromatin, iar citoplasma srac,
din care cauz hiperplaziile tumorale par a fi formate numai din nuclei. Stroma
este foarte redus.
Seminomul, tumora malign a parenchimului testicular, a fost identificat
sub dou forme: seminom intratubular i seminom de tip difuz.
Adenocarcinomul de prostat la cine evolueaz insidios i cu malignitate
crescut. Se dezvolt din epiteliul ductelor sau al acinilor. Se recunoate dup
hiperplaziile de celule rotunde sau ovale ce se dezvolt pe o fin tram
conjunctiv n lumenul alveolelor glandulare (tipul alveolar papilar).
Datele obinute de noi privind incidena tumorilor spontane la cine n
perioada investigat corespunde cu datele prezente din literatura de specialitate (1,
2, 3, 4, 5).
Gradul de malignitate al tumorii diagnosticate histopatologic nu se
coreleaz uneori cu aspectele macroscopice, mai evident n cazul tumorilor
maligne epiteliale, adenocarcinoame.
Datorit numrului mic de cazuri investigate, nu s-a putut stabili o relaie
direct ntre inciden i anumite rase.
Rezultatele comparative ale anchetelor epidemiologice efectuate n ultimii
30 de ani, arat o schimbare spectaculoas a localizrilor n diferite esuturi i
regiuni anatomice. Astfel, dac n anul 1968 tumorile pielii la cine erau cele mai
frecvente (28,1%), astzi acestea au fost depite de cele cu localizri mamare
care au atins procente ntre 57% (perioada 1979 1984) i 45% n perioada 1987
1989. Femelele sunt afectate ntr-un procent dublu fa de masculi datorit
incidenei mai mari a localizrilor mamare, uterine i vaginale (1, 4).

CONCLUZII

Histopatologic din totalul de 54 de tumori spontane la cine:


- 20 cazuri au fost diagnosticate la sexul mascul (37,03%);
- 34 cazuri au fost diagnosticate la sexul femel (62,96%).
Tumorile epiteliale benigne (adenoame, papiloame) par a fi mai numeroase
(54,54%) n timp ce tumorile mezenchimale benigne (fibroame, leiomioame) sunt
uor mai reduse (45,45%).

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n cazul tumorilor benigne se nregistreaz o evoluie asemntoare la cele


dou sexe. Cu alte cuvinte, frecvena apariiei tumorilor benigne nu este
influenat de sex ci, n principal, de vrst.
n cazul tumorilor maligne raportul dintre tumorile epiteliale (carcinoame,
adenocarcinoame) i tumorile mezenchimale (sacroame, fibrosarcoame) este net
n favoarea tumorilor epiteliale (81,25%).
Frecvena tumorilor maligne n cazul femelelor crete odat cu vrsta pn
n jurul vrstei de 12 ani. n general, tumorile diagnosticate au fost ncadrate n
categoria neoplaziilor maligne (59,25%).

BIBLIOGRAFIE

1. Baba A.I., 2002 - Oncologie comparat, Ed. Academiei Romne, Bucureti, 49-87.
2. Dorn C.R., 1976 Epidemiology of canine and feline tumors, J.Am.Anim. Hosp.
Assoc., 12, 307-334.
3. Jubb, K.V.F., Kennedy P.C., Palmer N., 1993 Patologhy of Domestic Animals, vol.
I-III, Acad. Press, New-York.
4. Manolescu N., Bolte S., 1991 Oncologie veterinar, vol. I i II, Ed. Ceres,
Bucureti, 9-51.
5. McGavin M.D., Carlton W.W., Zachary J.F., 2001 - Special veterinary pathology,
Ed. Mosby, USA.
6. Owen L.N., 1980 TNM clasification of tumours in domestic animals, OMS Geneva
7. Paul, I., 1990 - Morfopatologia aparatelor i a sistemelor organice, Bul. Inf. Soc.
Med,Vet. nr. 23-24, Bucureti.
8. Slauson D.O., Cooper B.J., 2002 - Mechanisms of disease, Ed. Mosby, USA.
9. Withrow, S.J., MacEwen, E.G., 2001 - Small animal clinical oncology, Ed. Saunders,
USA.

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MORPHOPATHOLOGICAL CHANGES IN THE


INFECTIOUS PERITONITIS IN CAT AND ASPECTS
RELATED TO THE HISTOPATHOLOGICAL DIAGNOSIS
OF THIS DISEASE
I. OLARIU1, M. COMAN1, AL. LAZU1,
S. LAZU2, A. STANCU1
1
U.S.A.M.V. Timioara
2
Bioveti Farm Timioara

Cat infectious peritonitis or feline infectious peritonitis (angeitis with


coronavirus or immune mediated coronoviral vasculitis) is a progressive
and mortal disease that affects domestic cats and also the wild free or under
captivity felines (1, 2, 7, 9).
This disease is determined by one or many coronaviruses antigenic
similar with the virus of swine transmissible gastroenteritis, canine
coronavirus and the virus 229E of human bronchitis (Padersen et al., 1978)
(10). Cats of all ages are receptive to this disease, especially those with the
age of 6 months - 2 years, with a 100% mortality in the case of the
perfectioned breeds and of the cats from breeding units. Its clinical signs
are anaemia, weakening, bad general status and peritonitis (3, 4, 7, 11, 12).
According to the way of infection and the immune status of the
infected cats, there are two forms or types of this disease: the effusive form
(moist, exudative) and the non-effusive form (dry, granulomatous) (1, 5, 6,
7, 10, 11).

Key words: cat, infectious peritonitis, lesions

MATERIALS AND METHODS

The researches were carried out during October 2005 February 2006,
through the necropsy of five cat bodies (four Birmanese and a common breed
one), with the ages between 6 months and 2 years, that presented gingival ulcers,
jaundice and abdominal distention, successive to a serious serofibrinous
peritonitis. We took samples from these bodies from lungs, cord, liver and
mesenter. The samples were fixed in formaldehyde solution 10%, processed
withthe help of the paraffin technique, sectioned at 6 micrometres and coloured
with the trichromic Masson method modified by V. Ciurea with methyl blue
(H.E.A.).
After the examination and interpretation of the preparations
histopathologically obtained, we took some microphotos to illustrate the most
specific histopathological changes.

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RESULTS AND DISCUSSIONS

Necropsically, within the peritoneal cavity we observed the presence o a


bad-senelling liquid, yelow-red, in a quantity of 800 ml, in which the rohole
gastrointestinal mass was incorporated serohemorrhagic peritonitis (fig. 1)
Lungs
Macroscopically, both lungs had increased volumes, weight and a mosaic-
like aspect, the pleura was tensioned, shiny and covered with fibrine on some
areas. The section surface was dry, mat, finely granular (rough), with colour
differences between the densed tissue and the bronchial apparatus, this one having
a lighter shade than the red-brown lung background (marmorate aspect) and a
positive docimasia the fibrinous pleurobronchopneumonia (fig. 2).

Fig. 1. Cadavru pisic- PIF (effusive form) Fig. 2. Fibrinous pleurobronchopneumonia -


Serohemorrhagic peritonitis PIF, effusive form

Microscopically, we observed the hyperemia of the alveolar capillars,


intraalveolar fibrino-haemorrhagic exudate, within the bronchic and peribronchic
lumen. The fibrino-haemorrhagic and leukocytary exudate is consisted of a fibrine
network in whose loops are present numerous hematia and/or leukocytes and
descuamated alveolar cells that removes completely the air from alveolas. There
are many areas with compensatory alveolar pulmonary emphysema adjacent to
the focuses of pulmonary compaction. These aspects are concordant to the
croupous pneumonia the stage of red hepatisation (fig. 3).
Cord
Macroscopically, in three cases, the pericardic sack was slightly relaxed
due to the serofibrinous exudate. The miocard was whitish-grey, not glistering,
with a mat aspect and friable consistence, and the right cord was flabby.
Microscopically, we observed fragmentations and dissociations of the
miocardic fibres; basophil granulations of various dimensions within the
sarcoplasm; plasmexodia, perivascular leuko- and erythrodiapedesis.

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Liver
Macroscopically, it had an increased volume and weight, the colour yellow-
ochre at surface and upon the section, and a friable consistence. The capsule was
tensioned, shiny and covered in some places with greish-yellow membranes of
fibrine, easily detachable fibrinous perihepatitis and hepatic steatosis (fig. 4).

Fig. 3. Croupous pneumonia the stage of


Fig. 4. Fibrinous perihepatitis and
red hepatisation - PIF ,effusive form. Col.
hepatic steatosis PIF, effusive form
HEA x 100

Fig. 5. Nodular mesenteritis-PIF - effusive Fig. 6. Pyogranuloma mesenteritis PIF


form Col. HEA x 200

Microscopically, we observed granulations and vacuoles optically empty


within the hepatocyte cytoplasm, the congestion of the centrolobular vein, of the
perilobular sanguine vessels and of those within the portobiliar space, the ectasy
of the sinusoid capillars; the presence of some optically empty areas of various
forms and sizes within the structure of the hepatic cordons hepatic necrosis.

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Mesentery
Macroscopically, we observed the presence of numerous nodular systems,
with the size of a millet grain, greish-yellow, uniformly distributed upon the
mesenteric surface nodular mesenteritis (fig. 5).
Microscopically, we identified mesothelial hyperplasia and neoformation
saguine capillars an rare nodules consisted of neotrophil cells, big monocyte cells,
plasmocytes and lymphocytes, which are delimited by a slight fibrilar
mesenchimal response pyogranuloma (fig. 6).

CONCLUSIONS

Infectious peritonitis (the exudative, moist or effusive form) was identified


necrospically and histopathologically in five domestic cats of various breeds and
with ages between 8 months and 2 years old.
Necrospically, we observed: gingival ulcers, jaundice and abdominal
distension; fibrinous pericarditis, protidic miocardosis and right cardiac dilatation;
fibrinous pleurobronchopneumonia the stage of red hepatisation; fibrinous
hepatitis, steatosis and miliar hepatic necrosis; nodular mesenteritis.
Histopathologically, we observed: fibrinous poli serositis (pleuritis disease,
pericarditis and perihepatitis); vascular, degenerative and necrotic changes of
various cardic-hepatic intensities and mesenteric pyogranuloma.
The histopathological diagnosis of infectious peritonitis, the effusive form,
was determined taking into account the serious vascular changes present within
the examined tissues and organs.

BIBLIOGRAHPY

1. Baba A.I., 1996 - Diagnostic necropsic veterinar, Ed. Ceres, Bucureti.


2. Ctoi C., 2003 Diagnostic necropsic veterinar, Ed. Academic Pres, Cluj-Napoca.
3. Green C.F., 1998 Infectious Disiases of the Dog and Cat. Ilt edition, W.B. Saunders
Company Philadelphia, London, Toronto, Montreal, Sydney, Tokyo.
4. Horzinek M. http:/www, newman veterinary.Com/F.I.P. Peritonitis, p.d.f (accesat
06.03.2006).
5. Jubb K.V.F., Kenedy P.C., Palmer N., 1993 Pathology of domestic Animals, vol. 1,
2, 3, Ed. Academic Press, San Diego, California, U.S.A.
6. McGavin M.D., Cartlon W.W., Zachary J.F., 2001 Special Veterinary Pathology,
Ed. Mosby, Inc, Missouri, U.S.A.
7. Moga Mnzat, R. Coordonator, 2005 Boli virotice i prionice ale animalelor, ed.
Brumar, Timioara.
8. Olariu-Jurca I., 2006 Diagnostic necropsic i medicin legal veterinar, Ed.
Brumar, Timioara.
9. Paul I., 1982 Diagnostic morfopatologic veterinar, Ed. Ceres, Bucureti.
10. Paul I. 1996 Etiomorfopatologie veterinar, vol. 1, Ed. All Bucureti.
11. Vasiu C., 2003 Viroze i boli prionice la animale. Ed. Neremia Napocal.
12. xxx http:/www. winnfelineheath. org/health/F.I.P. html (accesat 06.03.2006)

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EFECTUL PRODUCTIV AL UNUI PREBIOTIC


ASUPRA PERFORMANELOR PRODUCTIVE
LA PUII BROILER DE GIN
THE PRODUCTIVE EFFECT OF A PREBIOTIC OVER THE PRODUCTIVE
PERFORMANCES AT BOILER CHICKEN

D. SIMEANU, Gh. STAN, B. PSRIN


U.S.A.M.V. Iai

Pentru a stabili efectului productiv al prebioticului NUTRI-SURE


DW1 asupra puilor broiler de gin s-au format dou loturi experimentale
(E1 i E2), a cte 25 pui fiecare, i un lot martor (M) alctuit din 50 pui.
Puii din loturile experimentale au primit n apa de but 2 respectiv 2,5,
factor experimental, n perioada de via a puilor 1-21 zile i 1 respectiv
1,25 n perioada 22-35 zile.
Utilizarea acestui prebiotic la puii broiler de gin a avut efecte
benefice, n special n prima sptmn de via a puilor, determinnd
sporuri de producie mai mari cu 3,2-4,08% mai ridicate la loturile
experimentale fa de lotul martor. Consumul specific de hran a fost foarte
puin influenat, diferenele fiind nesemnificative. Starea de sntate a fost
corespunztoare, pierderile din efectiv nscriindu-se n limite normale
pentru aceast categorie de animale.

Key word: broiler chicken; prebiotic.

MATERIAL AND WORK METHOD

The experiment was carried out within the Nutrition and Alimentation
Biobasis from the Agricultural sciences and Veterinary Medicine Iasi on a total
number of 100 broiler hen chickens that were divided in three lots, of which a
witness lot (M) and two experimental lots. (E1 and E2) (tab.1).
The purpose of this experiment was to establish the influence of the
prebiotic NUTRI-SURE DW1 on the growth performances, the valorization of food
and health state of the hen broiler chickens HUBBARD FLEX.
The food administered was elaborated by FNC AviTop Tomesti, in three
periods: 1 14 days, the starting period, 15-28 days, the growth period and 29-35
days, the ending period.
The structure and the nutritive characteristics of the combined forages
administered are presented in tab. 2.
The experimented prebiotic is a mixture of formic, lactic and acetic acid,
that acts selectively against some bacteria like: Campylobacter and Salmonella,
reducing the pH from the stomach and the thin intestine, which determined a
good health state and a good valorization of food.
This preparation is not administered during the use of antibiotics.

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Table 1
Experimental schema

Lots Experimented period (days) Number of chickens /lot Experimental factor *


M 35 50 -
2 1-21 zile
E1 35 25
1 22-35 zile
2,5 1-21 zile
E2 35 25
1,25 22-35 zile
Note: * acidifying NUTRI-SURE DW1, administered in water..

Table 2
Structure and the nutritive characteristics of the combined forages

Raw Materials (%) Starting Growth Finishing


Maize 46,22 49,45 40,21
Soy groats 30,00 22,00 12,97
Full Fat soy 10,00 17,53 26,67
Maize gluten 5,00 4,50 4,00
Barley - - 10,00
Fish flour 3,72 - -
Molasses - 2,00 2,00
Monocalcic phosphate 1,65 1,74 1,46
Calcium carbonate 1,16 1,39 1,26
Wafolin 0,70 - -
Premix 0,50 0,50 0,60
Rhodimet 0,29 0,20 0,39
L-Lizin 0,24 0,14 0,03
Salt 0,18 0,27 0,18
Knob 50% 0,12 0,10 0,08
BioPlus 2B 0,10 0,05 -
Cycostat 0,06 - -
Cygro - 0,06 -
Kemzyme Ms dry 0,05 0,05 0,05
L-Treonin 0,02 - -
Sodium bicarbonate - 0,02 0,10
Nutritive characteristics
EM kcal/kg nc 3215,41 3249,00 3225,00
PB% 24,00 21,00 20,00
GB% 7,06 7,15 7,89
CB% 3,82 3,43 3,90
Ca 1,00 0,94 0,85
P disponibil % 0,50 0,45 0,42
Ca/P 2,00 2,10 2,02
L% 1,44 1,15 1,05
M+C% 1,09 1,06 0,96
Treonin% 0,93 0,79 0,75

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RESULTS AND DISCUSSIONS

The use of the product was benefic especially during the first period (1-7
days), when the weight increases were superior with 3,2% in the case of E1 lot
respectively, 4,08% in the case of E2 lot, compared to lot M (tab. 3)
Table 3
The evolution of body weight

The average weight The average Weekly %


Cumulated
Lots at the beginning of weight at the end daily gain comp.
increase (g)
the period (g) of the period (g) (g) to M
1-7 zile
M 42,35 164,32 17,42 121,97 -
E1 41,19 167,06 17,98 125,87 +3,20
E2 41,05 167,99 18,13 126,94 +4,08
7-14 zile
M 164,32 435,12 38,68 392,77 -
E1 167,06 432,58 37,93 391,39 -1,38
E2 167,99 436,92 38,41 395,87 +0,79
14-21 zile
M 435,12 842,73 58,23 800,38 -
E1 432,58 817,23 54,95 776,04 -3,04
E2 436,92 816,69 54,25 775,64 -3,09
21-28 zile
M 842,73 1329,67 69,56 1287,32 -
E1 817,23 1327,91 72,95 1286,72 -0,60
E2 816,69 1273,78 65,29 1232,73 -4,24
28-35 zile
M 1329,67 1856,49 75,26 1814,14 -
E1 1327,91 1866,21 76,90 1825,02 +0,60
E2 1273,78 1762,74 81,58 1803,79 -0,57

During the period 7-14 days, the results were different thus, the E1 lot
registered a decrease of weight with 1,38% while E2 lot registered an increase of
0,79% in contrast with the comparison lot.
During the period 14-28 the results from the experimental lots were inferior
to lot M, fact that indicates that in this period the product is not recommended.
Starting the 22nd day the use doses of the acidifying were reduced at half
(1% at E1 and 1,25% at E2).
In the last week, the E1 lot registered an increase of +0,6% compared to lot
M, while lot E2 had a decrease of 0,57%, compared to M, which can be
considered inconclusive.
We appreciate the fact that the product had a visible positive effect in the
first period of age having a stimulatory effect on the enzymatic secretion, but
especially on the gastric and pancreatic lipase synthesis.

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The food was well used in all the experiment lots, which was due to the
special quality of the combined forages used, the differences between the three
lots being insignificant (tab. 4).
Table 4
The food consume

The total Av.


Number of Average IC (kg %
consume of Daily
Lots chickens consume n.c./kg comp.
combined forage gain
/lot daily/head (g) increase) to M
(kg/lot) (g)
M 159,23 48 94,77 60,09 1,577 -
E1 83,21 25 95,10 60,45 1,573 -0,25
E2 79,25 24 94,34 59,74 1,579 +0,12

The health state was similar, the losses registered being found in the normal
limits for this category of birds, but we still noticed a highly increased mortality at
the witness lot compared to the two experimental lots (tab. 5).
Table 5
The evolution of discrepancies with the group

The number of chickens at


The no. of chickens of at Losses
Lots the beginning of the %
the end of the experiment (chickens)
experiment
M 50 48 3 6,25
E1 25 25 0 -
E2 25 24 1 4,00
Total losses 4%

CONCLUSIONS

We appreciate the fact that the influence of the acidifying was manifested
especially in the first week of age, when the increases were 3,2-4,08% higher at
the experimental lots compared to the witness lot.
The specific food consume was very little influenced by the NUTRI-SURE
DW1 prebiotic, the differences being significant.
The health state was similar, the collectivity losses being found in the
normal limits for this category of animals.

BIBLIOGRAPHY

1. Stan Gh., I.M. Pop, 1997 The alimentation and nutrition of animals, Junimea
Publishing House, Iasi.
2. Simeanu D., 2004 Biostimulators in the bird alimentation, Alfa Publishing House,
Iasi.
3. Stan Gh., D. Simeanu, 2005 - Animal nutrition, Alfa Publishing House, Iasi.
4. Stan Gh., D. Simeanu, 2005 The bird alimentation, Alfa Publishing House, Iasi.

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ASPECTE HEMATOLOGICE, BIOCHIMICE I


IMUNOLOGICE CONSECUTIV SUPLIMENTRII HRANEI
CU SEL-PLEX I ZN-BIOPLEX LA VACILE LACTANTE
THE HEMATOLOGICAL, BIOCHEMICAL AND IMMUNOLOGICAL ASPECTS TO
LACTATING DAIRY COWS FEEDED WITH ORGANIC ZINC AND SELENIUM

D. CURC1, A. LIONIDE2, ADriana ORANU3,


Ioana CONSTANTINESCU3, M. SAMARINEANU4,
M. CORNIL1
1
U.S.A.M.V., Bucureti
2
I.C.P.B.M.V., Bucureti
3
I.D.S.A., Bucureti
4
S.N. Institutul Pasteur

Recently trace minerals have been given more attention because of


their implication in several disorders that include heart disease, cancer,
early aging, malabsorption and immune disorders. Several elements play
important roles in controlling and catalyzing chemical reactions that are
vital for living organisms.
Essential trace elements are used for a multitude of purposes that
include being the backbone of energy exchange systems, controlling free
radicals and holding together proteins. Less specific clinical signs
indicating a lack of Selenium and vitamin E relate to unthriftiness of
livestock, including poor growth, diarrhea and susceptibility to disease. Are
also described the hematological and biochemical modifications in dairy
cows feeded with Sel-Plex and Zn-Bioplex. It was evaluated the efect of
feeding of organic sources of zinc (as Zn-Bioplex) and selenium ( as Sel-
Plex ) to lactating dairy cows on lymphocyte reactivity.
The experiment show, that feeding Sel-Plex increase
phytohemagglutinin lymphocyte stimulation index with 32,4%, Zn-Bioplex
with 56,7% and both Sel-Plex and Zn-Bioplex with 67,5%. Our results
demonstrate the positive influence of this supplemental zinc and selenium
sources to lactating dairy cows on hematological parameters and
lymphocyte transformation involved in cell-mediated immunity response.

Key words: Dairy cows, Zn-Bioplex, Sel-Plex, biochemical modification,


cell-mediated immunity

Zincul i seleniul sunt necesare att pentru buna cretere i dezvoltare a


plantelor, ct i pentru animale, orice celul conine o anumit cantitate din aceste
bioelemente ce intervin n numeroase procese metabolice, aflndu-se totodat n
interrelaii sinergice sau antagonice cu alte bioelemente (2, 6, 7, 8). Zincul are un
rol important n stimularea secreiei de insulin i implicit n reglarea
metabolismului glucidic, intervine n procesele de reproducie, a celui de protecie
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a vitaminei B1 i hormonilor TSH i LH, favoriznd fecundarea, regleaz


concentraia de vitamin A n plasma sanguin i n mobilizarea ei din ficat (4, 10,
11, 16, 17). Este implicat de asemenea, n sinteza keratinei, a acizilor nucleici din
piele, precum i n sinteza colagenului. Particip la reglarea echilibrului
hidroelectrolitic din organism, prin schimbul de cationi/anioni. Bovinele, n
general, i n special vacile pentru lapte au nevoie de zinc suplimentar n hran
pentru asigurarea unei funcii normale a organismului, fie n cazul unui aport
sczut de zinc din ingredientele hranei, fie datorit prezenei factorilor antagonici
care duc la scderea biodisponibilitii acestui bioelement (3, 9, 13).
Seleniu era considerat n prima jumtate a secolului XX, drept un agent
toxic, ce provoca tulburri ale membrelor i moartea animalului consecutiv
consumului de plante bogate n acest bioelement, ulterior pe baza cercetrilor
experimentale i a observaiilor clinice, a fost considerat drept un nutrient
esenial, fiind un component al enzimelor implicate n protecia antioxidant, n
metabolismul hormonilot tiroidieni etc. (18, 19, 20). Seleniu joac un rol
important n creterea tineretului i n fertilitate la bovine (12, 15), activnd
sinergic cu vitamina E, substituindu-se cu succes n prevenirea i tratamentul mai
multor entiti morbide care au la baz stresul oxidativ.
Activitatea enzimelor glutationperoxidazice (GSH-Px) depinde de nivelul
seleniului furnizat de hran, influennd mpreun cu vitamina E procesele
energogene respectiv sistemele enzimatice importante pentru respiraia celular
intervenind n stabilizarea membranelor lizozomale i al esuturilor bogate n
glutationperoxidaz. Dar, trebuie s se in cont de faptul c, seleniul are o aciune
antioxidant de 500 2000 ori mai mare dect vitamina E (1, 5).
Meninerea echilibrului antioxidant prooxidant n tractul digestiv, n
snge i esuturi prin utilizarea seleniului organic, este cheia prevenirii declinului
performanei productive i reproductive la animale, n condiii de stres asociate cu
sistemele de cretere intensiv (8, 14, 18, 19).
Sel-Plex-ul i-a dovedit avantajele, comparativ cu alte surse de seleniu,
deoarece furnizeaz acelai amestec de selenoaminoacizi aflai n ingredientele
naturale precum: grul, orzul, soia, porumbul etc.
Plecnd de la aceste considerente, s-au investigat efectele surselor organice
de zinc i seleniu (Bioplex Zn i Sel-Plex) prin suplimentarea hranei vacilor
pentru lapte, asupra unor indici hematologici, biochimici i imunobiologici (prin
aprecierea activitii limfocitelor T, cu ajutorul testului de transformare blastic,
considerat ca un indicator al dezvoltrii rspunsului imun).

MATERIAL I METOD

Cercetrile experimentale au fost efectuate un numr de 20 vaci pentru


lapte, rasa BNR, aflate n lunile a treia i a patra de lactaie, avnd o producie
medie de 24 l lapte/zi, iar starea de ntreinere fiind corespunztoare. Animalele

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au fost repartizate n 4 loturi a cte 5 vaci/lot, care au beneficiat de acelai regim


furajer. Suplimentarea cu zinc organic i respectiv seleniu organic s-a realizat prin
folosirea produselor comercializate de firma Alltech Romnia i anume: Bioplex-
Zn i Sel-Plex, astfel: lotul M (martor) care nu fost suplimentat cu cele 2
bioelemente; lotul E1 suplimentat cu 5 g Sel-Plex, echivalentul a 4,8 mg
seleniu/vac/zi; lotul E2 suplimentat cu 1,2 g Bioplex-Zn, echivalentul a 180 mg
Zn/vac/zi; lotul E3 suplimentat cu 5 g Sel-Plex (echivalentul a 4,8mg
seleniu/vac/zi) i 1,2 g Bioplex-Zn (echivalentul a 180 mg Zn/vac/zi).
Durata experimentului a fost de 45 de zile, la sfritul acestei perioade s-au
recoltat probe de snge pentru efectuarea investigaiilor de laborator i anume:
pentru examenul hematologic sngele a fost recoltat pe EDTA, iar determinrile
s-au fcut la un aparat automat Coulter-Counter, ACT 5 diff CP-Beckman;
investigaiile biochimice s-au efectuat prin utilizarea analizorului de biochimie
umed Eppendorff ECOM 1022, folosindu-se reactivii furnizai de Linear
Chemicals-Spania; s-au recoltat probe de snge pe heparin pentru determinarea
indicelui de stimulare (IS) a limfocitelor, folosindu-se testul de transformare
blastic a limfocitelor cu PHA (fitohemaglutinin).

REZULTATE I DISCUII

Rezultatele obinute n urma investigaiilor hematologice au evideniat


tendina de scdere a constantelor eritrocitare directe: eritremia, hemoglobinemia
i hematocritul. Scderile au fost marcate la lotul E1 suplimentat cu Sel-Plex; iar
la lotul E2 suplimentat cu Zn-Bioplex i lotul E3 suplimentat cu Sel-Plex + Zn-
Bioplex, scderile au fost mai reduse. n schimb, constantele eritrocitare derivate
au prezentat variaii n sensul creterii, fie al scderii nesemnificative.
Leucocitemia la vacile din lotul E1 a prezentat o cretere fa de vacile din lotul
martor cu 5,19%, iar la cele din lotul E2 creterea a fost nesemnificativ, n timp
ce la vacile din lotul E3 s-a constatat o scdere de 10,90%. Trombocitemia la
vacile din loturile experimentale a prezentat o scdere marcat la lotul E1
(21,50%); la E2 scderea a fost de 31,46%, iar la E3 scderea a fost doar de
30,30% (tabelul 1).
Valorile VSH-ului la 24 h au evideniat creteri marcate la vacile din lotul
experimentale, ndeosebi la lotul E2 unde creterea a fost de 95,10% fa de vacile
din lotul martor, urmat de vacile din lotul E1 unde creterea a fost de 82,51% i
E3 unde creterea a fost de 54,54% (fig. 1.).
La nivel biochimic s-au constatat modificri ale albuminemiei care a
prezentat creteri la vacile din loturile E2 i E3, n timp ce la E1 s-a evideniat o
scdere, n sens invers s-au modificat valorile globulinemiei (Tabelul 2). Uremia a
prezentat scderi vacile din lotul E1 i E3, n timp ce la vacile din lotul E2 s-a
remarcat tendina de cretere. Creatininemia a crescut semnificativ la vacile din
loturile experimentale comparativ cu cele din lotul martor, la E1 creterea a fost de
30,18%.

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55,8
60 52,2

50
44,2
40
28,6
30

20

10

0
Martor Suplimentat cu Suplimentat cu Suplimentat cu
Sel-Plex Zn-Bioplex Se si Zn

Fig. 1. - Modificrile VSH-ului dup 24 h, la bovinele crora li s-au administrat per os


seleniu i zinc, form organic.

Investigaiile privind coninutul unor bioelemente (calciu, magneziu,


fosfor), au evideniat creteri ale calcemiei n special la vacile din lotul E3
(83,52%), creteri ale magneziemiei la vacile din loturile E2 i E3, n timp ce la
vacile din lotul E1 scderea a fost de 50%. Raportul Ca/Mg a evideniat creteri la
vacile din loturile E1 i E3, n timp ce la cele din lotul E2 scderea a fost de
61,99%. Fosforemia a prezentat o tendin de cretere minim la vacile din
loturile experimentale.
La nivel enzimatic, s-a remarcat creterea GOT-ului la vacile din lotul E1 i
E3, n timp ce la E2 s-a constatat o uoar scdere. GPT-ul a prezentat tendina de
scdere la vacile din loturile E1 i E2 i uoar cretere la vacile din E3. Raportul
GOT/GPT evideniaz creterea marcat la vacile din loturile E1 i E3. GGT-ul
scade la vacile din loturile experimentale, ndeosebi la E3 unde scderea a fost de
45,46%. Fosfataza alcalin are tendina de scdere la vacile din lotul E2 i E3 n
timp ce la E1 se observ o uoar cretere de 6,62% (tabelul 3).
Rspunsul imun constituie n fapt o amplificare a funciilor celulelor
anticorpoformatoare, consecutiv unor modificri foarte reduse n echilibrul
homeostatic, ce debuteaz iniial prin reacii locale i apoi prin diseminarea
efectorilor, extinzndu-se progresiv n ntreg organismul.
Principalele celule implicate n rspunsul imun sunt cele prezentatoare de
antigen i limfocitele, procesul de amplificare fiind bidirecional: macrofagele
instruiesc limfocitele care prolifereaz i recruteaz mai multe macrofage n
vederea amplificrii fenomenelor i restaurrii homeostaziei (Tizard, 1992).

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Implicarea limfocitelor T n procesul de anticorpogenez se realizeaz prin


proliferarea clonal n prezena macrofagelor, care elaboreaz un al doilea semnal
i anume, interleukina 1 (IL-1), care este transmis prin contact direct celul la
celul sau chiar la distan. De aceea, proliferarea limfocitelor T se poate face i
n afara contactului cu macrofagele prin semnale constituite din mediatorii
solubili eliberai de ctre acestea, fiind cunoscute sub denumirea de limfokine, ce
particip alturi de semnalele elaborate de monocite, ce poart numele de
monokine.
Elaborarea acestor semnale poate fi modulat de echilibrul homeostatic al
organismului, care vizeaz n primul rnd aportul de nutrieni: aminoacizi,
glucide, lipide, vitamine i bioelemente. ntruct exist puine informaii
privitoare la modularea transformrii blastice a limfocitelor T la bovine sub
influena suplimentrii hranei cu Zn i respectiv Se sau ambelor, s-a efectuat
prezentul studiu care ofer informaii demne de luat n seam.
Loturile suplimentate cu Bioplex Zn i Sel-Plex au nregistrat o valoare
medie a indicelui de stimulare a limfocitelor mai ridicat comparativ cu lotul de
control M, astfel: n cazul lotului E1 s-a constatat o cretere a IS fa de lotul
martor cu 32,4%, n cazul lotului E2 cu 56,7 %, lotul E3 nregistrnd o valoare cu
67,5% mai mare dect lotul martor (fig.2).

23 24.8
25
19.6
20
14.8
15

10

0
Martor Suplimentat cu Suplimentat cu Suplimentat cu Se
Sel-Plex Zn-Bioplex si Zn

Fig. 2. Indicele mediu de stimulare a limfocitelor tratate cu PHA la vacile de lapte


suplimentate cu Bioplex Zn i Sel-Plex

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De asemenea, se constat c lotul E3 (suplimentat cu Bioplex Zn i Sel-


Plex) nregistreaz valoarea medie a indicelui de stimulare cea mai mare, 24,8 (cu
6,8% mai mare dect n cazul lotului E2 i cu 26% fa de lotul E1), urmat de lotul
E2 cu o valoare a IS mediu de 23,2 (cu 18,3% mai ridicat dect n cazul lotului
E1) i de lotul E1 cu o valoare medie a IS de 19,6.

CONCLUZII

1. Modificrile hematologice la vacile a cror hran a fost suplimentat cu


bioelemente, s-au caracterizat prin tendina de scdere a constantelor eritrocitare
directe, ndeosebi la lotul E1, n timp ce, constantele eritrocitare derivate au
prezentat variaii nesemnificative. Leucocitemia a prezentat o tendin de cretere,
n timp ce trombocitemia, o tendin de scdere.
2. Modificrile biochimice au prezentat tendina de cretere a calcemiei i
magnezemiei la loturile E2 i E3, n timp ce la lotul E1 s-a remarcat tendina de
scdere; variaiile fosforemiei au fost nesemnificative.
3. La nivel enzimatic s-a remarcat creterea GOT-ului, scderea GPT-ului
i fosfatazei alcaline.
4. Se remarc o cretere a indicelui de stimulare limfocitar indus de PHA:
cu 32,% n cazul lotului E1; cu 56,7% n cazul lotului E2; cu 67,5% n cazul lotului
E3 .
5. Rezultatele obinute n urma investigaiilor experimentale prin
suplimentarea hranei cu Sel-Plex i Zn-Bioplex, evideniaz influenele favorabile
asupra statusului indicilor hematologici, biochimici, precum i a imunitii
determinate prin aprecierea indicelui de stimulare limfocitar.

BIBLIOGRAFIE

1. Azizi E.S., Klesius P.H., Frandsen J.C., 1984 Effects of selenium on polymorpho-
nuclear leucocyte function in goats. American Journal of Veterinary Research,
45, 1715-1719.
2. Combs G.F. Jr., 1999 Chemopreventive mechanisms of Selenium. Medizinische
Klimik, 94 (Suppl. 3), 18-24.
3. Ghergariu S., 1981 Profilul metabolic la vaci; semnificaia clinic n stri de
infertilitate. Rev. de Creterea Animalelor, nr. 2.
4. Ghergariu S. i col., 1984 Indicatorii sanguini ai sntii hepatice la taurine.
Relaii morfofuncionale la vaci negestante, nelactante. Revista Creterea
Animalelor, nr. 7.
5. Gutzwiller A., 1998 Erythrocyte resistance to oxidative damage and leucocyte
capacity to reduce nitroblue tetrazolium in selenium deficient cattle. Zentralblatt
fr Veterin rmedizin Reihe A 45 (5), 271 278.
6. Harris B. Jr., 2003 The effcet of feeding zinc proteinate to lactating dairy cows.
University of Florida, Gainesville, USA.
7. Hemken R.W., Harmon R.L., 1998 Selenium for dairy cattle; A role for organis
selenium. Feed Componuder., October, 22 24.

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8. Kohrle J., Gartner R., 2000 Selenium in biology. Facts and medical perspectives.
Biol. Chem., 381, 849 867.
9. Lionide A., Curc D., Samarineanu M., 2005 Modificrile reactivitii limfocitare la
vacile de lapte consecutiv suplimentrii raiei cu forme organice de zinc i
seleniu. Lucrri tiinifice, seria C, XLVIII, 212 218.
10. Lowe J.A., 2003 An investigation into the metabolism of supplemental protected
zinc with reference to the use of isotopes. Gilbertson and Page Ltd., UK,
Nottingham University, Leicestershire, UK.
11. McDowel L.R., 1992 Minerals in Animal and Human Nutrition. Academic Press
Inc., Harcourt Brace Jovanovich Publishers, San Diego, CA, USA.
12. Mihai D., 1996 Boli de nutriie i metabolism la animale. Ed. Ceres, Bucureti.
13. Reffett J.K., Spears J.W., Brown T.T., 1988 Effect of dietary selenium on the
primary and secondary immune response in calves challenged with infectious
bovine rhinotracheitis virus. Journal of Nutrition, 118, 229 235.
14. Rotruck J.T., Pope A.L., Ganther H.E., Swanson A.B., 1973 Selenium:
biochemical role as a component of glutathion peroxidase. Science, 179, 588
590.
15. Schrauzer G.N., 2000 Selenomethionine: a review of its nutritional significance;
metabolism and toxicity. J. Nutr., 130, 1653 1656.
16. Spain Jim, Harris Barney Jr., 1997 Le rle du Zinc dans l`alimentation des
vaches laitires. Feedings Times, vol. 2, no. 1, p. 17.
17. Spears J.W., 2000 Micronutrients and immune function in cattle. Proc. Nutrition
Society, 59, 1 8.
18. Surai K.P., Surai P.F., Speake B.K., 2003 Antioxidant prooxidant balance in the
intestine. Food for though. Nutritional Genomics and Functional Foods, vol. 1,
51-70.
19. Surai P.F., 2002 Antioxidant protection in the intestine: a good beginning is half the
battle. Nutritional Biotechnology in the Feed and Food Industries, 301-321.
20. Underwood E.J., Suttle N.F., 1999 Selenium. In the Mineral Nutrition of Livestock,
CAB International, Oxon, 421 475.

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OBSERVAII PRIVIND VARIAIILE PROFILULUI


HEMATOLOGIC LA OVINE CU FASCIOLOZ
ON THE VARIATIONS OF THE HAEMATOLOGICAL PROFILE
IN OVINES WITH FASCIOLIASIS

Geta PAVEL
U.S.A.M.V. Iai
E. PURICE
CSV Vidra, jud.Vrancea

Haematological measurements on a number of 35 adult sheeps (3-6


years old) from a herd of ovines infested with Fasciola hepatica took place
in October 2005 when the beginning phase of the chronic fascioliasis
occurred. Results emphasized a small towards medium anaemia in all
investigated animals (Hb, Ht, E showed values under the inferior limit of
the species). The anaemic morphologic type proved by VEM and CHEM
modifications was variable: macrocytic hypochromic, of reactive type (with
reticulocyte within the peripheral blood); macrocytic normochromic, due to
the deficit hepatic synthesis of the transport protein B12; normocytic
normochromic, typically to an insufficient erythrogenese or to the lifetime
reduction of the erythrocytes.
Leukogram emphasized different leukocytary reactions: sometimes
moderate neutrophily with a left shift of the nuclear index, accentuation of
the lymphopoiesis with the presence of the lymphoblasts and active
lymphocytes in the peripheral blood; other times neutropenia with
lymphocytose; and in other cases eosinophilia. No leukocytosis occurred,
although the total number of circulating leukocytes showed values close to
the superior limit of the species, while the number of the trombocytes was
close to the inferior limit of the normal values.

Key words: fascioliasis, sheeps, anaemia, leukocytary reaction, eosinophilia

La ovine, strile anemice sunt relativ frecvente. In afar de anemiile


careniale, datorate aportului insuficient de proteine, vitamina B12, Fe, Cu, Co,
ovinele prezint anemii posthemoragice n cursul invaziilor parazitare, cnd se
produc leziuni traumatice ale vaselor de snge (9).
Fascioloza ovin nu ntmpltor este denumit i anemie verminoas sau
anemie de iarn. Boal cu caracter sezonier, evolueaz acut n perioada
septembrie-octombrie, sau cronic, cu o faz de debut n septembrie-noiembrie, o
faz de stare n decembrie-ianuarie i o faz final n februarie-martie (3, 8).
Anemia se manifest n toate formele de evoluie i n toate stadiile
fasciolozei. Fasciolele tinere, aflate n migraie, sap tunele n parenchimul
hepatic producnd hemoragii apoi fibroz. Formele adulte de Fasciola hepatica,

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ajunse n canalele biliare, devin hematofage, absorbind sngele din capilarele


pereilor canalelor biliare. In plus, se consider c fasciolele elimin cantiti mari
de prolin, care provoac, printre altele, i perturbri ale sintezei hemoglobinei
(8).
Scopul prezentei lucrri este evidenierea modificrilor parametrilor
hematologici la ovine dintr-un efectiv la care infestrile i reinfestrile cu
metacercari s-au produs masiv n ultimii ani, datorit ploilor abundente ce au
permis dezvoltarea gazdelor intermediare (gasteropode din familia Limneidae).

MATERIAL I METOD

Cercetrile s-au efectuat pe un numr de 35 oi dintr-un efectiv de ovine


expus infestrii cu Fasciola hepatica. S-au selectat pentru investigaii animale
adulte, femele, n vrst de 3-6 ani, care au exteriorizat semnele clinice
caracteristice fazei de debut a fasciolozei cronice (octombrie, 2005). De la aceste
animale s-au recoltat probe de snge pentru efectuarea determinrilor
hematologice. S-au investigat eritrocitele, leucocitele i trombocitele,
determinndu-se hematocritul (Ht), concentraia hemoglobinei (Hb), numrul de
eritrocite (E), constantele eritrocitare derivate (VEM, HEM, CHEM), procentul de
reticulocite, numrul de leucocite totale circulante (L), formula leucocitar (N, Eo,
Ba, Lf, Mo), numrul de trombocite i morfologia elementelor figurate.
Determinrile cantitative s-au efectuat prin metodele clasice de laborator
(hemocitometrie, metoda colorimetric Sahli, microhematocrit), iar explorarea
calitativ s-a realizat prin analiza citomorfologic a frotiului de snge colorat May-
Grnwald-Giemsa i prin coloraia vital cu albastru de briliant crezil pentru
determinarea reticulocitelor (4, 5, 6).

REZULTATE I DISCUII

Rezultatele obinute la ovinele investigate sunt prezentate n tabelele 1, 2


i 3.
Din analiza rezultatelor se constat c toate animalele cu fascioloz au
prezentat valori ale numrului de eritrocite, hematocritului i hemoglobinei sub
sau la limita inferioar a speciei. Majoritatea autorilor raporteaz pentru oile
adulte sntoase urmtorile valori normale: E 9-15 x106/l (cu o medie de 12); Hb
9-15 g/dl (cu o medie de 11,5), Ht 27-45 % (cu o medie de 35) (4, 7, 10). La
ovinele investigate de noi, am obinut valori sczute, dar nu drastic sczute, astfel
c dup gravitate anemia este uoar sau medie, cu valori pentru Hb ntre 7,4 i
10,0 g/dl, pentru Ht ntre 22 i 34 %, iar pentru E ntre 5,03 i 6,82 x106/l.
Valorile constantelor eritrocitare derivate au permis diagnosticarea tipului
morfologic al anemiei pentru fiacare caz n parte. Unele animale au prezentat
anemie macrocitar hipocrom, altele tipul de anemie macrocitar normocrom,
iar altele tipul de anemie normocitar normocrom.

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Anemia macrocitar hipocrom este de tip reactiv, fiind nsoit i de


apariia reticulocitelor n sngele periferic. Anemia macrocitar normocrom este
cauzat de o deficien de sintez hepatic a proteinei de transport pentru vitamina
B12, necesar procesului de eritrogenez medular. Tipul normocitar normocrom
de anemie diagnosticat s-ar putea datora depresiei medulare, dar n bolile hepatice
cauza cea mai probabil a anemiei este reducerea duratei de via a hematiilor.
Acest lucru a fost dovedit de Ogunrinade i Anosa, 1981, care au constatat c
timpul de njumtire a hematiilor marcate cu 51Cr s-a redus (128,8 ore) la oile
infestate cu Fasciola gigantica, comparativ cu timpul de njumtire al
hematiilor la oile de control (288 ore). Dup ali autori, n general, bolile hepatice
se asociaz cu o reducere a sintezei eritropoetinei de origine hepatic,
diminundu-se i n acest mod eritropoeza medular (4, 10).
Analiza frotiurilor de snge a evideniat modificri morfologice ale
eritrocitelor (anizocitoz, poikilocitoz, prezena anulocitelor, prezena
acantocitelor) i o redus reticulocitoz (ntre 0,8% i 1,6%), care are semnificaie
diagnostic, avnd n vedere faptul c la ovine, ca i la celelalte rumegtoare,
reticulocitele nu se elibereaz n sngele periferic n condiii fiziologice (1, 4, 5,
9).
Rezultatele privind seria leucocitar la ovinele cu fascioloz n stadiu de
debut arat absena eozinofiliei sau prezena unei creteri a procentului de
eozinofile numai pn la valoarea de 20 % din totalul leucocitelor. Literatura de
specialitate prevede valori normale ale numrului de eozinofile la ovinele adulte
de 0-10% (cu o medie de 5%) (Jain, 1986), iar pentru fascioloza uman, este
prevzut o eozinofilie accentuat (80-90%) (Gherman, 1990) (2, 4). Acest aspect
nu s-a nregistrat la cazurile de fascioloz ovin studiate.
Numrul de trombocite la oile cu fascioloz nu au prezentat variaii fa de
valorile nregistrate n literatura de specialitate (2,5-7,5 x105/l, cu o medie de 4,0
x105/l, citate de 4). Astfel valorile gsite de noi pentru numrul de trombocite au
variat ntre 2,7 i 4,2 x 105/l, majoritatea valorilor fiind sub media normal a
speciei.
Numrul de leucocite totale s-a situat ctre limita superioar, peste valoarea
medie citat n literatura de specialitate (8000/l, cu limite de 4000-12000/l) (4).
Astfel, la oile cu fascioloz investigate, numrul leucocitelor totale a variat ntre
9,1 i 11,6 x 103/l; se poate vorbi de o tendin ctre leucocitoz sau de o
leucocitoz moderat la unele animale.
Analiza formulei leucocitare i a numrului absolut de N tinere, N mature,
Eo, Ba, Mo, Lf, la oile cu fascioloz investigate, evideniaz un tablou extrem de
variabil. Unele animale (10 cazuri) au prezentat neutrofilie moderat, cu o uoar
deviere spre stnga a indicelui nuclear, raportul N:Lf fiind de 1,1:1, uor n
favoarea neutrofilelor, aspect care se abate de la normal (la ovine, raportul N:Lf
fiind de 1:2). Aadar, la animalele investigate s-a surprins o reacie inflamatorie
acut, probabil cu localizare hepatic. Alte 15 cazuri de ovine cu fascioloz au

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prezentat neutropenie i limfocitoz, raportul N:Lf fiind de 1:7,2, iar alte 10


cazuri au evideniat un raport N:Lf normal (1: 2,5), dar au prezentat eozinofilie
(numrul de Eo ajungnd la 20%).
Studiul citomorfologic al frotiului de snge a evideniat pentru seria
leucocitar prezena de neutrofile tinere, chiar metamielocite la unele cazuri,
alturi de neutrofile mature hipersegmentate, eozinofile vacuolizate, limfoblati i
limfocite active (cu granulaii, cu bazofilie accentuat).

CONCLUZII

1. In fascioloza ovin cronic, tipul morfologic de anemie este variabil: fie


macrocitar hipocrom, de tip regenerativ (cu prezena unei modeste
reticulocitoze), fie macrocitar normocrom, caracteristic deficienei de vitamina
B12 (n cazul fasciolozei fiind implicat o deficien de sintez hepatic a proteinei
de transport a vitaminei B12 ), fie normocitar normocrom, datorat reducerii duratei
de via a hematiilor sau a sintezei hepatice a eritropoetinei.
2. Migraia fasciolelor tinere prin esutul hepatic i fixarea formelor adulte
n canaliculele biliare genereaz o reacie inflamatorie local, reflectat sistemic,
fie prin neutrofilie, cu devierea spre stnga granulocitar, dar i limfocitar
(prezena limfoblatilor n sngele periferic), fie prin neutropenie i limfocitoz,
fie prin eozinofilie.
3. La oile cu fascioloz investigate n luna octombrie, eozinofilia nu a fost
constant i nu a a depit valoarea de 20 % n formula leucocitar.
4. Numrul leucocitelor totale circulante, la oile cu fascioloz, a prezentat
valori peste media normal a speciei, ctre limita superioar, iar numrul
trombocitelor a nregistrat valori sub medie, ctre limita inferioar a speciei.

BIBLIOGRAFIE

1. Ghergariu, S., AL. Pop, L. Kadar, Mariana Spnu, 2000 Manual de laborator
clinic veterinar, Ed.All, Bucureti.
2. Gherman, I., 1990 Dicionar de parazitologie, Ed. tiinific, Bucureti.
3. Iacob, Olimpia, 2002 Diagnosticul bolilor parazitare la animale, Ed. Ion Ionescu de
la Brad, Iai.
4. Jain, N.C., 1986 - Schalm's Veterinary Hematology. Lea-Febiger, Philadelphia.
5. Kerr, M.G., 1989 - Veterinary Laboratory Medicine. Clinical Biochemistry and
Haematology. Blackwell Scientific Publication, Oxford.
6. Kondi, V., 1981 - Laborator clinic de hematologie. Ed. Medical, Bucureti.
7. Manolescu, N. i colab., 1999 Tratat de hematologie animal, vol I, Ed. Fundaiei
Romnia de mine, Bucureti.
8. Mitrea, I.L., 2002 Boli parazitare la animale, Ed Ceres, Bucureti.
9. Prvu Gh., 1992 Supravegherea nutriional-metabolic a animalelor, Ed.Ceres,
Bucureti.
10. Straus, J.H., 1998 Anemia. Polycythemia, in Merck Veterinary Manual, Eight
Edition, Merck-Co., Inc. Whitehouse Station, N.J., USA.

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STUDIU COMPARATIV AL REACTIVITII VASCULARE


ARTERIALE LA MAI MULTE SPECII DE MAMIFERE:
1. REACTIVITATEA MUSCULATURII NETEDE
ARTERIALE LA AGENII VASOCONSTRICTORI
S. BECHEA-CHIRIAC1, W. BILD2,
Veronica NASTASA3, I. HULIC2
1
U.S.A.M.V. Iai
2
Academia Romn, Filiala Iai - L.F.E.A.
3
U.M.F. Iai

Vascular reactivity is one of the three pillars on which lies the


regulation of arterial pressure in living organisms. Arterial pressure is one
of the main determinants of the activity state of various organs and systems
both in healthy and in pathologically-altered states. The present study aims
toward identifying similarities and differences between the resistance
arteries belonging from various mammal species that are most involved in
veterinary practice: rats, cats, dogs and horses. The arterial fragments
prelevated from animals dead due to various clinical and traumatic
conditions unrelated to vascular pathology were normalized using a newly-
introduced system of quantification, the force index system. This has been
calculated using the wet-weight parameter and the force generated after
administration of various pharmacological agents that cause
vasoconstriction. The artery fragments were fitted in organ baths using the
Krebs-Henseleit saline, thermostated at 37 C and bubbled with a mixture
of 95% O2 and 5%CO2. Vascular endothelium was either kept or removed
using gentle rubbing with moist filter paper. Control of endothelial removal
was made both functionally, using carbachol (synthetic derivative of
acetylcholine) and microscopically, after testing. The force generated was
measured using isometric force transducers coupled to a computerized
acquisition system. The pharmacological vasoconstricting agents used were
phenylephrine (synthetic derivative of epinephrine), KCl (potassium
chloride 40-80 mM, as depolarizing agent) angiotensin II, and vasopressin.
The results were statistically investigated using the t-test and ANOVA
testing. The preliminary results show a dependence of the force generated
an the amount of muscle present in the various species from which the
arteries were taken, a specifically increased response of feline-derived
arteries to angiotensin and a specifically increased response of canine-
derived arteries to vasopressin. These results will be used as controls for
further testing in various pathological conditions and for various other
pharmacological agents used in the therapy of vascularly-induced
pathological states.

Key words: vascular, reactivity, arterial, vasoconstrictor agent..

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Studiul de fa dorete s investigheze modificrile de reactivitate vascular


arterial care ar putea fi implicate n patologia diverselor specii de animale
ntlnite mai frecvent n practica veterinar. S-a urmrit realizarea unei
investigaii comparative a reactivitii vasculare la segmente arteriale similare din
punct de vedere histologic i funcional, recoltate de la diverse specii de mamifere
ntlnite mai des n patologia veterinar.
Investigarea mecanismelor ce stau la baza reglrii tonusului arterial i a
metabolismului fibrei musculare netede arteriale s-a bazat n ultimele dou
decenii pe foarte cunoscutul model al traductorilor izometrici i al preparatului
inelar de diverse artere. Vasul arterial de elecie folosit pentru astfel de
investigaii este aorta de obolan, deoarece ntrunete majoritatea condiiilor de
stabilitate, accesibilitate, disponibilitate i control unor investigaii de ncredere.
La aceasta se adaug criteriul de pre, care nu este deloc un element de pierdut din
vedere.
Totui, dei acest model este arhicunoscut, obolanul nu este un model
perfect din punct de vedere al modelrii cardiovasculare, care s fie similar cu
omul i cu att mai puin cu alte mamifere. Perioada de-a lungul creia se ntind
aceste studii ce au implicat acest model experimental ncepe acum jumtate de
secol [3].
Astfel, s-au folosit fragmente de artere recoltate de la cini, pisici i cai,
care au fost supuse investigrii experimentale comparativ cu inele de aort
toracic standardizate, recoltate de la obolani Wistar.

MATERIAL I METOD

Reactivitatea inelelor arteriale a fost msurat att n termeni de for


absolut, msurat ca indice de for (fora n mN dezvoltat de preparat
raportat la greutatea acesteia n mg) ct i ca reacie relativ, fa de un martor
standardizat. De asemeni, acolo unde a fost posibil (n funcie de disponibilitatea
preparatelor) s-au efectuat curbe doz/efect ce au implicat majoritatea
substanelor vasorelaxante i vasoconstrictoare cunoscute i bine caracterizate
farmacologic.
Studiul comparativ a fost fcut pe artere similare din punct de vedere a
dimensiunilor, care s fac parte din segmentul de rezisten, n spe ramuri din
artera coronar gastric sau mezenterica superioar, care au avut dimensiuni
similare: lungime maxim: 2 mm, = 1 mm, greutate 10-15 mg. Cuantificarea
forei de contracie a fost realizat n N/mg greutate umed [4]. Piesele de organ
au fost prelevate de la Clinica Medical i de la Clinica Chirurgie a Facultii de
Medicin Veterinar de la animale moarte n urma unor afeciuni fr implicare
vascular sau supus eutanasiei n condiiile legii [8].
Dup disecie, vasele au fost exsanguinate, splate n soluie de ser
fiziologic, secionate n fragmente de 5-10 cm i apoi au fost plasate n ser Krebs-
Henseleit (preparat conform formulei) i transportate in timp maxim, 30 minute la
locul experimentului.

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Fragmentele de aort au fost fixate prin intermediul unei serfine metalice


pe fundul bailor de organ izolat, inelul tensionndu-se prin intermediul vernierelor
mrcilor tensiometrice la o tensiune iniial de 100 mN.
Atunci cnd caracteristicile experimentale au cerut-o, endoteliul vascular a
fost ndeprtat prin frecare blnd cu hrtie de filtru umed. Prezena endoteliului
vascular funcional a fost verificat farmacologic (folosind carbachol) i prin
microscopie direct.
Inelele de arter au fost montate n bi de organ coninnd 4 ml de ser
fiziologic Krebs-Henseleit (compoziie (mM): NaCl 118; KCl 4.7; CaCl2 2.52;
MgSO4 1.64; NaHCO3 24.88; KH2PO4 1.18; glucoz 5.55), termostatate la 37C i
barbotate cu carbogen (amestec de oxigen 95% i CO2 5%).
Pentru nregistrarea contraciilor musculaturii netede vasculare s-au folosit
traductori izometrici de for conectai la un sistem computerizat de achiziie a
datelor.
Preparatele au fost lsate la echilibrat timp de 60-90 minute, sub o
tensiune de repaos de 100 mN.
Inele de arter au fost apoi precontractate cu fenilefrin (10-7 10-6)M i K+
(40-70 mM) i tratate cu carbachol (10-6M) pentru eliberarea de NO endotelial [6].
Magnitudinea absolut a contraciilor a fost de 175 25 mN pentru fenilefrin (10-
6
M) i K+ (40-70 mM).

REZULTATE I DISCUII

Fenilefrina este un agonist -adrenergic sintetic folosit n practica medical


ca midriatic, decongestionant nazal, iar n practica medical veterinar, rareori, ca
agent cardiotonic.
Receptorul su este reprezentat de adrenoceptorul 2 A.
Acesta este un receptor ubiquitar la nivelul sarcolemei muchiului neted,
cuplat cu proteina G. Efectul su se realizeaz prin intermediul inhibrii adenilat
ciclazei prin aciunea proteinelor G de tip q. Acestea, odat activate, stimuleaz
activiatea fosfolipazei C, stimulnd eliberarea de IP3 i DAG. Acetia acioneaz
ca mesageri secunzi ce mediaz eliberarea de Ca2+ intracelular cu efect direct
asupra contraciei musculare, i accesor, activeaz PKC [4].
Aciunea sa principal se realizeaz la nivelul receptorilor 2-adrenergic la
nivelul musculaturii netede arteriale, indiferent de localizare, producnd
vasoconstricie.
Administrarea de fenilefrin produce un efect stabil, puternic i
reproductibil la toate preparatele arteriale. De asemeni, s-a preferat substana
respectiv datorit faptului c fiind sintetic, este complet hidrosolubil i stabil
n soluie de-a lungul mai multor sptmni (fig. 1).
Datorit disponibilitii substanei respective, s-a folosit ca martor de
contracie n toate experimentrile ulterioare.

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2000
1900
1800
1700
Forta (mgF)

1600
1500
1400
1300
1200
1100
1000
7500 8000 8500 9000 9500 10000 10500 11000 11500 12000
timp (secunde)

Fig. 1 Aspectul tipic al contraciei fenilefrinice a unei artere din segmentul de rezisten
(Phe momentul administrrii, w splarea preparatului cu ser proaspt)

S-a realizat o curb doz efect la fenilefrin la preparate cu endoteliul


vascular pstrat ct i ndeprtat (fig. 2).
Investigarea efectelor substanelor vasoconstrictoare s-a realizat att ca
apreciere a indicelui de for (F/gu) ct i ca apreciere a contraciei raportat la
contracia indus de fenilefrin 10-6 M, care a fost considerat contracie de
referin (100%).

4
IF - mN/mg

Endotelizat
3
Dezendotelizat
2

0
-10 -9 -8 -7 -6 -5 -4
-1
log conc Phe

Fig. 2 - Curba doza-efect la fenilefrina - grup control

Indicii de for la vase endotelizate recoltate de la animale fr afectare


vascular.
obolan IF = F/gu = 2,5 gf /5 mg 25 mN/5 = 5
Pisic IF = F/gu = 3,1 gf/ 5mg = 6,2
Cine IF = F/gu = 3,4 gf / 5mg = 6,8
Cal IF = F/gu = 4,8 gf/5mg = 8,16

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unde: F reprezint fora dezvoltat de preparat, exprimat n mN (N x 10-3)


iar gu reprezint greutatea umed, exprimat n miligrame de esut.
Dup cum se poate observa n fig. 3, fora preparatelor de inel arterial
recoltate de la animalele luate n studiu a fost din ce n ce mai mare, pe msur ce
animalul de la care s-a recoltat fragmentul de vas a fost mai mare. Ca urmare,
responsivitatea adrenergic s-a dovedit a fi direct proporional cu cantitatea de
muchi neted vascular prezent la nivelul preparatelor.

10
9
8
IF (mN/mg tesut umed)

6 sobolan
pisica
5
caine
4
cal
3
2
1
0
Inele arteriale endotelizate

Fig. 3 - Variaia indicelui de for al inelelor arteriale la administrarea de Phe 10-6M

Efectul fenilefrinei la nivelul preparatelor dezendotelizate: dup cum se


poate observa din fig. 4, dezendotelizarea a avut un efect de cretere a reactivitii
la fenilefrin cu valori cuprinse ntre 16 % (obolan), 9,25% (pisic), 8,39 (cine)
i 9,27 % la cal. Aceasta este n concordan cu rezultatele din literatur i cu
ateptrile noastre conform rezultatelor anterioare.

12

10
IF (mN/mg tesut umed)

8
sobolan
pisica
6
caine
cal
4

0
1 2 3 4
inele arteriale dezendotelizate

Fig. 4 Variaia indicelui de for al inelelor arteriale dezendotelizate la administrarea de


Phe 10-5 M

n fig. 5 se prezint graficul cumulativ ce rezult din ambele seturi de date.


Se poate observa c efectul de stimulare a rspunsului contractil este mai redus la
arterele provenite de la animalele mari (cal, cine), ceea ce poate sugera o
implicare mai puin semnificativ a reglrii tonusului arterial bazal de ctre
catecolamine n condiiile arterelor de rezisten ce prezint mase musculare
foarte mari.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

16

14
sobolan
12

IF (mN/mg tesut umed)


pisica
10 caine
cal
8
Linear (sobolan)
6 Linear (pisica)
Linear (caine)
4
Linear (cal)
2

0
1 2 3 4 5 6 7 8

Fig. 5 - Rezultate cumulative (endotelizat-dezendotelizat), la care s-au adugat liniile de


regresie ale variaiilor ntre contracia preparatelor endotelizate i a celor dezendotelizate

CONCLUZII

Din rezultatele prezentate mai sus se pot trage urmtoarele concluzii:


Reactivitatea -adrenergic la toate speciile luate n studiu este similar,
dependent de doz.
Singurele diferene semnificative observate sunt cele cantitative,
determinate posibil de reprezentarea cantitativ a muchiului neted vascular la
nivelul diverselor tipuri de artere luate n studiu.
Raportndu-ne la datele histologice putem spune c, raportndu-se la
procentul de muchi neted, rezultatele se nscriu n limitele normalitii.

BIBLIOGRAFIE

1. Armstead W.M., Lippton H.L., Hyman A.L., Kadowitz P.J., 1984 - Analysis of
adrenergic responses in the mesenteric vascular bed of the cat; evidence that
vascular beta 2-adrenoceptors are innervated. Can J Physiol Pharmacol.
62(12):1470-8.
2. Bechea Chiriac S., Serban D.N., 2001 - Mechanisms involved in the vascular
action of phenamyl, Lucr. St. vol. 44., USAMV Iai, p. 151-159;
3. Beznak M., 1956 - Hemodynamic changes following aortic constriction in normal and
in hypophysectomized rats. - Can J Biochem Physiol., 34(4):791-8.
4. Guimares S., Moura D., 2001 - Vascular Adrenoceptors: An Update - Pharmacol
Rev. 53: 319-356.
5. Haulica I., Bild W., Mihaila C.N., Ionita T., Boisteanu C.P., Neagu B., 2003 -
Biphasic effects of angiotensin (1-7) and its interactions with angiotensin II in rat
aorta. - J Renin Angiotensin Aldosterone Syst. 4(2):124-8
6. Jiang H., Halayko A.J., Rao K., Cunningham P., Stephens N.L., 1991 -
Normalization of force generated by canine airway smooth muscles. - Am J
Physiol. 260(6 Pt 1):L522-9.
7. Moncada S., Palmer R.M.J., Higgs E.A., 1991 - Nitric oxide: physiology,
pathophysiology, and pharmacology. Pharmacol. Rev., 43: 109-142
8. * * * - LEGE nr.471 din 9 iulie 2002 Buletinul Oficial al Romniei, privind aprobarea
Ordonantei Guvernului nr. 37/2002 pentru protectia animalelor folosite n scopuri
stiintifice sau n alte scopuri experimentale.

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CLINICAL, ULTRASONOGRAPHICAL AND CYTOLOGIC


ASPECTS OF PERITONEAL EFFUSION IN FELINE
SUSPECTED OF INFECTIOUS PERITONITIS
ASPECTE CLINICE, ECOGRAFICE I CITOLOGICE ALE LICHIDULUI DE PUNCIE
LA FELINE SUSPECTE DE PERITONITA INFECIOAS

V BOGHIAN, Gh. SOLCAN, Geta PAVEL,


M. MUSTEA, R. SAUCIUC
U.S.A.M.V. Iai

Au fost examinate 14 cazuri de motani i pisici care prezentau


manifestri clinice de peritonit: febr intermitent, inapeten, slbire,
dispnee, comportament inhibat, i semnele fizice ale coleciei lichide
peritoneale. Lichidul de puncie peritoneal era incolor sau uor glbui,
vscos, uor coagulabil i cu coninut ridicat de proteine (5,51,5 g/dl). n
frotiul colorat panoptic s-au observat numeroase fagocite mari (neutrofile
i macrofage), limfocite i uneori hematii. La examenul ecografic s-au
observat modificri de ecogenitate ale ficatului i rinichiului (zone de
hiperecogenitate) i prezena de lichid n cavitatea peritoneal.

Cuvinte cheie: peritonita felin, semne clinice, ecografie,


citologia lichidului peritoneal

The Feline infectious peritonitis (PIF) is an infectious disease of viral origin


of the feline, produced by a Corona virus and characterized by general troubles,
weakness and peritonitis. Clinical symptoms are the results of the vascular and
per vascular alterations. As a consequence it is produced the extravasation of
plasma and the accumulation of this intracellular or in serous cavities (wet form
of evolution) and without cavitar overflowing (dry form of evolution). These
aspects make the clinical manifestation to be varied from case to case. Taking in
consideration that the clinical symptoms are polymorphous we tried to appreciate
which are the constant clinical aspects in peritonitis, and correlated with the
echographic aspects and the cytology of the peritoneal liquid, can be used to put
diagnosis (1, 2).

MATERIAL AND METHOD

The researches were made on 14 animals, males and females, of different


ages, clinical suspected of Infectious Feline Peritonitis (PIF). Using the clinical
examination were appreciated the constant aspects of the clinicinal symptoms.
Using the ultrasound examination were noticed the echostructure modification at
the level of the parenchymatos organs from the abdominal cavity, especially of
the liver and kidneys, and the presence or the absence of the peritoneal liquid

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collection. Related to the presence or the absence of the peritoneal liquid, the
investigations were completed with the sampling of the liquid from peritoneal
puncture and is exanimate from organoleptic point of view, morphological smear
stained May Grnwald Giemsa, and protein concentration using the direct
refractometric method.

RESULTS AND DISCUSSIONS

The constant clinical sings were: intermittent fever, inappetence,


progressive weakness, dyspnoea, inhibitive behavior, and the physical sings of the
peritoneal liquid collection in exudative form (wet form) of evolution of the
disease (fig. 1).

A B
Fig. 1. - A- Inhibitive behavior; B - Physical signs of the peritoneal liquid collection

In the dry form of the disease, clinical aspects were varied and not
characteristic, with predominant clinical sings depending on the localization of
the visceral lesions (hepatic, renal, medullar, encephalic granuloma or
hydrocephalus). The lesions of the organs from the abdominal cavity were
observed by ultrasonographic examination.
At the ultrasonographic examination the echostructure alter of the liver,
such as milliary zones of hyperechogenity and liquid collection inside the
peritoneal cavity were observed in exssudative form (fig. 2 and fig. 3).
The peritoneal liquid, present only in the exudative form of evolution of the
disease, was clear or yellowish, very viscous and rich in proteins; the total
proteins had the value of 5,51,5 g/dl. On the smear made after
ultracentrifugation of the liquid stained May Grnwald Giemsa were noticed red
cells and numerous leucocytes: big phagocytes, neutrophiles and lymph cells,
aspects characterizing the inflammatory liquid (fig. 4 and fig. 5).

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A B
Fig. 2. A-B: Liver with hiperechogenity zones and the presence of the liquid inside the
peritoneal cavity

A B
Fig. 3. A - The liquid inside the peritoneal cavity (perirenal); B - Liver with
hyperechogenic structure

A B
Fig. 4. A-B: Peritoneal liquid with red cells and numerous leucocytes (col.MGGx900)

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Universitatea de tiine Agricole i Medicin Veterinar Iai

A B
Fig. 5. - Peritoneal liquid: A - big phagocytes and erytrophagocitosis; B - neutrophiles,
lymph cells, red cells (col.MGGx1500)

CONCLUSIONS

The symptoms of peritonitis in cats were: intermittent fever, progressive


weakness, inhibitive behavior, and the physical signs of the peritoneal liquid
collection in the exudative form of evolution of the disease.
In the dry form of evolution of the disease the most important clinical signs
were: intermittent fever, weakness and hyperechogenic lesions of the liver.
At the ultrasonographic examination were noticed hiperechogenity milliary
zones on liver and the presence of the liquid inside the peritoneal cavity, very
useful for the diagnosis.
The peritoneal liquid was clear or yellowish, and very viscous, rich in
proteins (5,51,5 g/dl).
Cytological exam of peritoneal liquid revealed numerous leucocytes (big
phagocytes, neutropyiles and lymphocytes).

BIBLIOGRAPHY

1. Horzinek M., 2001 - Unravellig the Secrets of Feline infectious peritonitis-Sotal,


World Small Animal Veterinary Association World Congress Proceedings.
2. Moga-Mnzat R., 2005 - Boli virotice i prionice, Ed. Brumar, Timioara, p. 323-328.
3. Perianu T., 1997 - Boli infecioase ale animalelor, viroze (vol. II), Ed. Fundaiei
Chemarea, Iai, p. 150-151.
4. Solcan Gh., Boghian V., Rollin F., 2005 - Patologie i clinic medical veterinar,
Ed. Ion Ionescu de la Brad, Iai.
5. Solcan Gh., Luminia Diana HRICU, Bechea Chiriac S.I., Boghian V., 2003 -
Paraclinical, and ultrasonographic aspects in some hepatobiliary diseases of
dogs and cats, Bull. USAMV Cluj Napoca, seria Med. Vet., 60, p. 315-318.

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POST-PARTUM KETONEMIA DETERMINED BY BETA-


HYDROXYBUTYRATE ON HOLSTEIN FRIZE DAIRY
COWS
CETONEMIA POSTPARTUM PRIN DETERMINAREA BETAHIDROXIBUTIRATULUI LA
VACILE PENTRU LAPTE HOLSTEIN FRIZ

V. BOGHIAN
U.S.A.M.V. Iai

La un lot de 115 vaci pentru lapte, aflate n primele 3 luni de


lactaie, s-a determinat -hidroxibutiratul seric. S-au diagnosticat 12 vaci
cu valori crescute ale -hidroxibutiratului, ceea ce reprezint 10,43% din
efectiv. Din acestea, 10 vaci (8,69% din efectiv) au avut valori ale -
hidroxibutiratului ce caracterizeaz cetoza subclinic i 2 vaci (1,73% din
efectiv) au avut valori ale -hidroxibutiratului ce caracterizeaz cetoza
clinic. n funcie de starea fiziologic a animalelor, cele 12 cazuri pozitive
s-au diagnosticat astfel: 3 (25,0%) n primele 2 sptmni postpartum, 5
(41,66%) n sptmnile 2-4 postpartum, 3 (25,0%) n sptmnile 4-6
postpartum, i 1 caz (8,33%) n sptmnile 6-8 postpartum. Dup a aptea
sptmn de lactaie nu s-au mai ntlnit cazuri cu concentraii serice ale
-hidroxibutiratului de peste 14,4 mg/dl, ceea ce confirm faptul c vacile
aflate n primele 6 sptmni de lactaie au un risc mai mare de a dezvolta
cetoza comparativ cu cele aflate dup aceast perioad.

Cuvinte cheie: cetoza, vaci pentru lapte, -hidroxibutirat, diagnostic

Ketosis is as a metabolic state characterized through the transitory increase


of ketonic bodies concentration in organism. Taking in consideration this fact,
ketosis can be diagnosed determining the ketonemia. The -hydroxybutyrate
represents 78% from the total of ketonic bodies, is more stabile and can be easily
determined. At the same time is a usual metabolite at cows. It is physiologic
formed at the level of the luminal wall through the metabolisation of the butyric
acid (volatile fatty acid) contained by the fodder ration, or has endoluminal origin.
It also synthesized at the liver level. Because of this reason, at some
concentration, it is considered a physiologic metabolite on milk cows.
The level of the serous concentration of -hydroxybutyrate of over 1400
mol/l characterizes subclinical ketosis, and the concentration of over 2600
mol/l characterizes sub clinical ketosis. To change the measure unit of -
hydroxybutyrate to mg/dl is by multiplying the value mol/l with 0,0103. This
number represents the molecular mass of -hydroxybutyrate which is 103 divided
to 10.000, meaning the difference from g/l and mg/dl.(14000,0103=14,4
mg/dl). (2, 5).

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MATERIAL AND METHOD

The researches were made on a herd of 115 postpartum cows whit an


average production of milk of 29 l/day. On serum testes taken from the entire herd
at once, the -hydroxybutyrate was determined using the biochemical
semiautomatic analyzer EOS 880 PLUS and specific kit. The obtained results
were interpreted related to the value of -hydroxybutyrate and the physiological
state of the cows (the number of days which passed after giving birth).

RESULTS AND DISCUSSIONS

The dates obtained at the -hydroxybutyrate determination are presented in


table 1 and fig. 1.

35
30
25
20
15
10
5
0
0 20 40 60 80

BHB (mg/dl)

Fig. 1. The post-partum values of the -hydroxybutyrate on a herd of 115 dairy cows

Were diagnosed 12 cows, representing 10,43% of the total exanimated


herd, with the value of serum -hydroxybutyrate over 14,4 mg/dl. From these, 2
cows had the value of -hydroxybutyrate which characterizes the clinical ketosis
(1,73% from the examinated herd) and 10 cows had the value of -
hydroxybutyrate which characterizes subclinical ketosis (8,69% from the
delaminated herd).
41,66% (5 cases) were diagnosed in the 2-4th post-partum week. In the first
2 post-partum weeks were diagnosed 3 cases, representing 2,60% from the total
examinated herd, or 25% from the 12 positive cases. Same situation happened in
4-6th post-partum week. In the period 6-8th post-partum week was diagnosed only
one positive case, and in 8-10th post-partum week and 10-12th week there was not
diagnosed any positive case. To add, in these weeks the average value of serum -
hydroxybutyrate was less in comparison with the beginning of the nursing period.

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These dates confirm the fact that the cows being in the first 6 weeks of
postpartum have a higher risk to develop ketosis in comparison with the cows
being after this period. The cows which are in the 2-6th post-partum week period
have the higher risk to develop subclinical ketosis.
The weekly dynamics of the subclinical ketosis by determining -
hydroxybutyrate is presented in figure number 2.

0
I II III IV V VI VII VIII IX
Vaci cu cetoz subclinic (n)

Fig. 2.- The numerical post-partum incidence on cows with the value of -hydroxybutyrate
over 14,4 mg/dl

The most cases, meaning 4, were diagnosed in third post-partum week.


After the 7 th postpartum week there were not discovered any cases with serum -
hydroxybutyrate concentration over 14,4 mg/dl. It is also specified that the
examiated herd was formed from cows in first 75 postpartum days.
Was also noticed that the period in which the serum value of -
hydroxybutyrate is over the limit 14,4 mg/dl is sometimes very short, reason
which leads to the fact that some of the cows which will develop subclinical
ketosis have small serum concentration of -hydroxybutyrate, when they are
tested for the diagnosis of ketosis.
The serum concentration of aceto-acetate and -hydroxybutyrate represents
the relative total (without acetone) of the kenotic bodies. The value of the serum
concentration of these two kenotic bodies summing up over the level of 9 mg/dl is
a sign for subclinical ketosis. The summing up value of 9 mg/dl for the subclinical
ketosis is smaller than the one of the -hydroxybutyrate alone, of 14,4 mg/dl, due
to the fact that -hydroxybutyrate can have also endoluminal origin by consuming
fodder with accentuated butyric fermentation. At the level of the luminal wall,
through the metabolisation of the butyric acid (volatile fatty acid) contained by
the fodder ration or with endoluminal origin, there are being formed small

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quantities of -hydroxybutyrate. In this case -hydroxybutyrate is not related to


the hepatic ketogenesis; it is not formed from aceto-acetate. This is why some
serum concentration of -hydroxybutyrate, without aceto-acetate are considered
to be physiological at the milk cow. When at the serum concentration of -
hydroxybutyrate is added also the one of the aceto-acetate, it is obtained the
relative value of the total ketonemia, which suggest a hepatic ketogenesis,
considered to be pathological over a certain level.

CONCLUSIONS

From 115 examinated cows, 12 representing 10,43% of the herd had


increased values of -hydroxybutyrate.
10 cows (8,69%) had value of -hydroxybutyrate characterizing the
subclinical ketosis, and two cows (1,73%) had value of -hydroxybutyrate
characterizing clinical ketosis.
Related to the physihological state of the animals, those 12 positive cases
were diagnosed as follows: 3 (25,0%) in first two post-partum weeks, 5 (41,66%),
in 2-4 th post-partum week, 3 (25,0%) in 4-6 th post-partum week, and one case
(8,33%) in 6-8 th post-partum week.
After the 7 th postpartum week there not diagnosed any cases with serum
concentration of -hydroxybutyrate over 14,4 mg/dl, this confirming the fact that
the cows being in the first 6 postpartum weeks have a higher risk to develop
ketosis in comparison with the cows being after this period.

BIBLIOGRAPHY

1. Boghian V., Hagiu N., 2003 - Valoarea diagnostic a testelor de profil metabolic n
cetoza subclinic a vacilor pentru lapte, Lucr. t. Med. Vet., Ed. Ion Ionescu de
la Brad Iai, vol., 46 (5), pag. 262-265.
2. Duffield T.F., Kelton D.F., Leslie K.E., Lissemore K.D., Lumsden J.H., 1997 - Use
of test day milk fat and milk protein to detect subclinical ketosis in dairy cattle in
Ontario, Can. Vet. J., vol. 38 (11), pag. 713-718.
3. Geishauser T., Leslie K., Tenhag J., Bashiri A., 2000 - Evaluation of eight cow-
side ketone tests in milk for detection of subclinical ketosis in dairy cows, J. Dairy
Sci., vol. 83 (2), pag. 296-299.
4. Holtenius P., Holtenius K., 1996 - New aspects of ketone bodies in energy
metabolism of dairy cows, A review. J. Vet. Med., vol. 43 (10), pag. 579 587.
5. Oetzel G.R., 2001 - Ketosis and hepatic lipidosis in dairy herds, Preconvention
Seminar 8: Dairy herd problem investigations strategies, American Association of
Bovine Practitioners 34 th, Annual Convention, sept. 1112, Vancouver BC.

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EFECTELE DEFICITULUI HIDRIC PRELUNGIT LA


CERVIDEE, PE PERIOADA DE IARN
EFFECTS CONCERNING THE PROLONGED HIDRIC DEFFICIENCY IN STAGS, IN
WINTER PERIOD

Iuliana CODREANU, N. DOJAN, M. CODREANU


U.S.A.M.V. Bucureti
Ioana CONSTANTINESCU
I.D.S.A., Bucureti

The stag evolution like fellow of a population it is influence all the


time by the hereditary and natural factors.
Between these factors, part of it is not depends of population density,
this factor are: humidity, temperature and light.
The other factors that depend of population density that influence the
distribution and population level are: forage, water, shelter.
The aim of this study is to realize the biochemical the biochemical
profile of a lot of stags who it hast been secure the drink water for about 2-3
weeks in a winter months with snow and a lot of stags who it has been
secure the drinking water.
Comparing the results, it has been notice a increased of the seric
urea for about 4-5 times over the normal value of the species and also a bad
health conditions.

Att cadrul socio-economic al rii, ct i nivelul sczut al cercetrii i


educaiei cinegetice - i al neglijrii acestora - pe considerentul c cervideele din
mediul silvatic al Carpailor Orientali evolueaz natural, pozitiv, mai bine ca
oriunde n Europa, au condus la o serie de incertitudini referitoare la activitatea de
administraie i de gestiune a fondurilor cinegetice [5].
Factorii care influeneaz evoluia cantitativ i calitativ a cervideelor din
Carpaii Orientali sunt concurena acerb i mortalitile numeroase. Evoluia
cervideelor, ca indivizi ai populaiilor, are loc sub influena nentrerupt a aciunii
factorilor de mediu i a ereditii [1].
nsi ereditatea trebuie acceptat ca un rezultat al adaptrii speciei la
condiiile de mediu n care a evoluat.
Dintre aceti factori, o parte sunt independeni de densitatea populaiei
(mortalitatea cervideelor crescnd sau scznd n funcie de intensitatea i durata
aciunii lor) cum ar fi: umiditatea, temperatura, lumina i pot deveni, n unele
situaii, limitativi pentru rspndirea cervideelor [4].
Ceilali factori sunt dependeni de densitatea populaiei, influennd
distribuia spaial i nivelul populaional: hrana, apa, adpostul, linitea,
competiia intra- i interspecific, prdarea i parazitismul [4].

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MATERIALE I METODE

Investigaiile de ordin clinic s-au efectuat pe perioada 6-25 ianuarie 2005,


pe 22 de indivizi de aceeai vrst (1,6 ani), din cadrul Farming Group
Transilvania SRL, cu sediu n Reghin, judeul Mure, crora proprietarul nu le-a
asigurat ap de but, considernd c, fiind lun de iarn, acetia i vor asigura
necesarul de ap din zpad.
Dup dou sptmni, proprietarul a observat modificari grave ale strii de
sntate a cervideelor. n urmtoarele zile, 5 animale au decedat.
Astfel, proprietarul a hotrt s recolteze probe de snge de la animalele
rmase n via pentru efectuarea unor examene de specialitate.
Investigaiile biochimice au fost realizate prin metodele clasice, n
Laboratorul de Biologie molecular i Biochimie din cadrul Institutului de
Diagnostic i Sntate Animal Bucureti.
Investigaiile biochimice s-au realizat pe probe de ser sanguin prin
metodele titrimetrice clasice, curent efectuate n laboratoarele de specialitate
descrise [2,3].
n realizarea i aprecierea profilului biochimic la cervidee am considerat
oportun i semnificativ realizarea urmtoarelor investigaii:
- constituenii principali ai profilului proteic (protein total, albumine,
globuline), uree, creatinin;
- constituenii principali ai profilului mineral (Ca, Mg, P).
Studiile privind homeostazia unor factori umorali corelai cu starea de
sntate a cervideelor, s-a efectuat pe un numr de 22 de indivizi, mprii n 2
loturi, dup cum urmeaz:
- lotul LA lot martor - alctuit din 10 cervidee, crora li s-a asigurat
ap de but;
- lotul LB - alctuit din 12 cervidee, crora nu li s-a asigurat ap de but
(n perioada de iarn).

REZULTATE I DISCUII

n urma analizei comparative ale valorilor medii obinute n urma


examenelor biochimice, la cele dou loturile de cervidee, sunt redate , pentru
fiecare element de profil investigat, n tabelele 1-3.
n urma investigaiilor efectuate la cele dou loturi de cervidee, s-a
constatat existena unor diferene nesemnificative ntre valorile medii ale proteinei
totale, ele ncadrndu-se n limitele normale ale speciei (5,6-9,9 g/dl), n cazul
ambelor loturi.
De asemenea, i n cazul valorilor medii ale albuminelor i globulinelor s-a
constatat existena unor diferene nesemnificative din punct de vedere statistic
(p>0,05), ceea ce demonstreaz faptul c principalii constituieni ai profilului
proteic nu sunt afectai (modificai) n cazul cervideelor cu deficit hidric sever
(tabelul 1).

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Tabelul 1
Valorile medii ale proteinei totale, albuminelor i globulinelor la lotul martor (LA)
i respectiv la lotul de cervidee cu deficit hidric sever (LB)

Protein total Albumine Globuline


Lotul Raport A/G
(g/dl) (g/dl) (g/dl)
LA 7,16 3,18 3,98 0,80
LB 6,21* 2,64* 3,56* 0,74*

* p >0,05 diferene nesemnificative

Valoarea medie a ureei n cazul lotului de cervidee cruia nu i s-a asigurat


ap de but, a fost de 123,2 mg/dl (cu limite cuprinse ntre 83 mg/dl i 138
mg/dl), distinct semnificativ mai mare din punct de vedere statistic (p<0,01), fa
de valorile medii ale lotului LA (format din animale sntoase).
ntre valorilor medii ale creatininei, s-a constatat existena unor diferene
nesemnificative (p>0,05), n cazul celor dou loturi de cervidee (tabelul 2).
Tabelul 2
Valorile medii ale ureei i creatininei la lotul martor (LA) i respectiv la lotul
de cervidee cu deficit hidric sever (LB)

Uree Creatinin
Lotul
(mg/dl) (mg/dl)
LA 27,2 1,13
LB 123,2*** 1,23*

* p >0,05 diferene nesemnificative


*** p <0,01 diferene distinct semnificative

Se poate remarca faptul c, la indivizii cu privare hidric sever (LB), se


nregistreaz constant creteri severe ale ureei serice pe fondul hemoconcentraiei
i deshidratrii, modificri care au fost invariabil corelate cu prezena semnelor de
retenie azotat (insuficien renal acut) de tip progresiv agravant.
Cortegiul clinic aferent insuficienei renale acute, a constituit dominanta
clinic la indivizii privai de ap (LB).
Creterea nesemnificativ a valorii medii a creatininei serice i ncadrarea
ei n limitele normale ale speciei, n cazul indivizilor cu deficit hidric sever, indic
o agresiune renal de tip acut, fr instalarea unor veritabile alterri morfologice
la nivelul substratului filtrant renal.
Dezechilibrele n plan umoral cu consecine detrimentale sistemice, au stat
la baza agravrii progresive a strii clinice pn la faza de insuficien renal de
tip decompensat.
Valorile medii ale calciului i fosforului au fost de 11,1 mg/dl i respectiv
de 7,2 mg/dl la lotul de cervidee sntoase i de 12,5 mg/dl i 6,17 mg/dl la lotul
de cervidee crora nu li s-a asigurat ap de but.

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Diferenele dintre aceste valori medii, n cazul celor dou loturi de


cervidee, sunt nesemnificative din punct de vedere statistic (p>0,05) i se ncadrau
n limitele caracteristice speciei (9-12 mg/dl pentru calciu i 5,7-10 mg/dl pentru
fosfor), citate de literatura de specialitate (tabelul 3).
Tabelul 3
Valorile medii ale calciului, magneziului i fosforului seric la lotul martor (LA)
i respectiv la lotul de cervidee cu deficit hidric sever (LB)

Ca Fosfor Mg Vitamina E
Lotul Calcium Phosphor Magnesium Vitamin E
(mg/dl) (mg/dl) (mg/dl) (/ml)
LA 11,1 7,2 2,8 1,8
LB 12,5* 6,17* 3,02* 2,29*

* p >0,05 diferene nesemnificative

De asemenea, diferenele dintre valorile medii ale magneziului i vitaminei


E, la cele dou loturi de cervidee au fost nesemnificative, ncadrndu-se i acestea
n limitele normale ale speciei (1,8-3 mg/dl pentru magneziu i de 1,2-2 /ml
pentru vitamina E).

CONCLUZII

Analiznd comparativ rezultatele obinute n urma examenului biochimic la


lotul de cervidee la care s-a asigurat ap de but (lot martor) i la lotul de cervidee
cu deficit hidric sever (cruia nu i s-a asigurat ap de but pe considerentul c,
fiind iarn, animalele i asigur necesarul de ap din zpad), se pot desprinde
urmtoarele concluzii:
n cazul celor dou loturi de cervidee, ntre valorile medii ale principalilor
constituieni ai profilului proteic nu exist diferene semnificative, ele ncadrndu-
se n limitele caracteristice speciei.
Valorea medie a ureei serice la lotul de cervidee crui nu i s-a asigurat ap
de but pe perioada de iarn, a fost de 123,2 mg/dl (cu limite cuprinse ntre 83
mg/dl i 138 mg/dl), constatndu-se o cretere distinct semnificativ (p<0,01), fa
de lotul martor.
Creterile severe ale ureei serice pe fondul hemoconcentraiei i
deshidratrii la indivizii cu privare hidric sever, au fost corelate cu prezena
semnelor de retenie azotat (insuficien renal acut) de tip progresiv agravant
i au stat la baza insuficien renal de tip decompensat.
Diferenele dintre valorile medii ale creatininei, n cazul celor dou loturi
de cervidee, au fost nesemnificative (p>0,05), ncadrndu-se n general ntre
limitele caracteristice speciei.

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Meninerea valorii medii a creatininei ntre limitele normale ale speciei


indic, n cazul indivizilor cu privare hidric, o agresiune renal de tip acut, fr
instalarea unor veritabile alterri morfologice la nivelul substratului filtrant renal.
ntre valorile medii ale principalilor constituieni ai profilului mineral
(calciu, magneziu, fosfor), s-a constatat existena unor diferene nesemnificative
din punct de vedere statistic (p>0,05).

BIBLIOGRAFIE

1. Bentley A., 1999 An Introduction to Deer of Australia, Bunyip, Sydney


2. Dojan N., 2000 Fiziologia animalelor domestice, Printech, Bucureti
3. Dojan N., 2003 Compendiu de fiziologie animal, Printech, Bucureti
4. Fanning L., 2001 Mountain Majesty, Mill Ponn Press, USA
5. Nesterov V., 1987 Bolile vnatului, Ceres, Bucureti, 1987.

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DATE DE PROFIL METABOLIC (BIOCHIMIC SANGUIN)


LA CERVIDEE, N FUNCIE DE VRST
ASPECTS CONCERNING METABOLIC PROFILE (SERIC BIOCHEMICAL)
IN STAGS, DEPENDING ON AGE

Iuliana CODREANU, N. DOJAN, M. CODREANU


U.S.A.M.V. Bucureti
Ioana CONSTANTINESCU
I.D.S.A., Bucureti

The stag had obtained richly the quality of the Romanian symbol
save that nourishment but also for the national and world trophy value.
The generally health status in stags can be proper evaluate only
after the realising of the hematological and biochemical blood analyses,
and after that, the obtained results must be compared with the normal
values, registered or described for that breed. In many cases the blood
constituents - components of the biochemical profile - are helpful in the
evaluation of the body status.
The aim of this study is to realise the biochemical profile, containing
the major components of the specific profile in stags depending on age.
Are described and compared the obtained results after the
investigations of these biochemical parameters in young animals (females
and males) 7 months, and adults (2,5 years).

In pdurile de munte principalul vnat este cerbul, cu o greutate medie de


240-250 kg la masculi, care populeaz sectorul oriental al Carpailor pn n zona
curburii precum i unele zone din Carpaii Meridionali i Munii Apuseni pn n
Podiul Trnavelor.
Efectivul de cervidee calculat este de circa 30.000 de exemplare i prin
vntoare se pierd, anual, 1-2 mii de exemplare.
ngrijortor este faptul c n ultimii 8-10 ani numrul cerbilor s-a redus de
cca 10 ori [5].
Disprut din alte ri europene, la noi n ar cerbul este nc un vnat de
foarte mare importan.
Este unanim recunoscut c n Carpaii Orientali ai Romniei habiteaz, cei
mai puternici cerbi din Europa [5].
Cerbul i-a dobndit, pe deplin, calitatea de emblem a vnatului romnesc,
att ca surs de hran, dar mai ales pentru trofeele impozante, de valoare naional
i chiar mondial [1, 4].

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MATERIALE I METODE

n realizarea i aprecierea profilului biochimic la cervidee am considerat


oportun i semnificativ realizarea urmtoarelor investigaii:
- constituenii principali ai profilului proteic (protein total, albumine,
globuline);
- constituenii principali ai profilului mineral (Ca, Mg, P);
- constituenii principali ai profilului enzimatic: transaminazele serice -
GOT (transaminaza glutamicoxalacetic), GPT (transaminaza
glutamic-piruvic), GGT (gammaglutamiltrans-feraza), FA (fosfataza
alcalin).
Investigaiile biochimice au fost realizate prin metodele clasice, n
Laboratorul de Biologie molecular i Biochimie din cadrul Institutului de
Diagnostic i Sntate Animal Bucureti.
Investigaiile biochimice s-au realizat pe probe de ser sanguin prin
metodele titrimetrice clasice, curent efectuate n laboratoarele de specialitate
descrise [2, 3].
Investigaiile de ordin clinic s-au efectuat pe loturi de cervidee cu vrste
diferite, din cadrul Farming Group Transilvania SRL, cu sediu n Reghin, judeul
Mure.
Studiile privind homeostazia unor factori umorali corelai cu starea de
sntate a cervideelor s-a efectuat pe un numr de 20 de indivizi (femele i
masculi), mprii n 2 loturi, dup cum urmeaz:
- lotul L1 - alctuit din 10 indivizi n vrst de 7 luni;
- lotul L2 - alctuit din 10 indivizi n vrst de 2,5 ani.

REZULTATE I DISCUII

Analiza comparativ a rezultatelor obinute n urma examenelor biochimice


la cele dou loturile de cervidee, alctuite din indivizi n vrst de 7 luni i
respectiv 2,5 ani, sunt redate n tabelele 1-3.
Valorile medii ale calciului i fosforului au fost de 13,99 mg/dl i respectiv
de 6,71 mg/dl la lotul de cervidee n vrst de 7 luni i de 12,7 mg/dl i 6,90
mg/dl la lotul de cervidee n vrst de 2,5 ani (tabelul 1).
Tabel 1
Valorile medii ale calciului, magneziului i fosforului seric la loturile de cervidee
n vrst de 7 luni i respectiv 2,5 ani

Ca Fosfor Mg Vitamina E
Lotul Vrsta
(mg/dl) (mg/dl) (mg/dl) (/ml)
L1 7 luni 13,99 6,71 2,58 1,78
L2 2,5 ani 12,7* 6,90* 2,75* 2,18*
* p >0,05 diferene nesemnificative

Diferenele dintre valorile medii ale celor dou loturi sunt nesemnificative
(p>0,05) i se ncadrau n limitele caracteristice speciei (9-12 mg/dl pentru calciu

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i 5,7-10 mg/dl pentru fosfor), citate de literatura de specialitate [1, 5]. De


asemenea, diferenele dintre valorile medii ale magneziului i vitaminei E, la cele
dou loturi de cervidee au fost nesemnificative, ncadrndu-se i acestea n
limitele caracteristice speciei (1,8-3 mg/dl pentru magneziu i de 1,2-2 /ml
pentru vitamina E).
La cele dou loturi de cervidee, s-a obinut aceeai valoare medie a
proteinei totale, ea ncadrndu-se n limitele normale ale speciei (5,6-9,9 g/dl)
oferite de literatura de specialitate [1].
ntre valorile medii ale albuminelor i globulinelor s-a constatat existena
unor diferene nesemnificative din punct de vedere statistic (p >0,05) ntre cele
dou loturi cu vrste diferite, valori care se ncadrau de asemenea, n limitele
caracteristice speciei (tabelul 2).
Tabel 2
Valorile medii ale proteinei totale, albuminelor i globulinelor la loturile de cervidee
n vrst de 7 luni i respectiv 2,5 ani

Protein total Albumine Globuline


Lotul Vrsta Raport A/G
(g/dl) (g/dl) (g/dl)
L1 7 luni 6,55 2,88 3,67 0,78
L2 2,5 ani 6,54* 3,05* 3,49* 0,88*
* p >0,05 diferene nesemnificative

Analiza comparativ a activitii transaminazelor serice GOT (ASAT) i


GPT (ALAT) n funcie de vrst la cervideele investigate, a evideniat creteri
constante ale activitii GOT-ului, odat cu naintarea n vrst (tabelul 3).
Tabel 3
Valorile medii ale activitii transaminazelor serice i fosfatazei alcaline la loturile
de cervidee n vrst de 7 luni i respectiv 2,5 ani

GOT GPT F.A.


Lotul Vrsta
(UI) (UI) (U/l)
LNI 7 luni 14,9 30,4 51,7
LNS 2,5 ani 30,3** 22,7* 52,6*
* p >0,05 diferene nesemnificative
**p< 0,05 diferene semnificative

Se poate aprecia faptul c activitatea seric a GOT-ului este crescut la o


valoare medie/lot de 30,3 UI (cu limite cuprinse ntre 19-48 UI), cretere
semnificativ din punct de vedere statistic (p<0,05), ce poate fi explicat pe baza
efortului i rolului metabolic deosebit, inclusiv la nivel hepatocitar, la vrst mai
naintat.
Variaiile GPT-ului n cadrul aceluiai lot (ntre valori de 11 UI i 48 UI),
precum i diferenele nregistrate ntre cele dou loturi de cervidee (fr
semnificaie statistic deosebit), indic o relevan mai redus a acestuia ca
indicator biochimic sanguin, inclusiv al funciei hepatocitare.

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Valorile activitii fosfatazei alcaline, att n cadrul aceluiai lot, ct i prin


comparaie ntre loturile investigate, precum i variaiile reduse nregistrate,
denot faptul c este un parametru ce sufer flucuaii nesemnificative, n funcie
de grupa de vrst.

CONCLUZII

Analiznd comparativ rezultatele obinute n urma examenului biochimic la


cele dou loturi de cervidee, alctuite din indivizi n vrst de 7 luni i respectiv
2,5 ani, se pot desprinde urmtoarele concluzii:
ntre valorile medii ale principalilor constituieni ai profilului mineral
(calciu, magneziu, fosfor) la loturile de cervidee investigate, s-a constatat
existena unor diferene nesemnificative din punct de vedere statistic (p>0,05).
Valorile medii ale principalilor constituieni ai profilului proteic se ncadrau
n limitele caracteristice speciei, constatndu-se existena unor diferene
nesemnificative (p>0,05) n cazul celor dou loturi.
Odat cu naintarea n vrst a cervideelor, se constat o cretere
semnificativ din punct de vedere statistic (p<0,05) a activitii GOT-ului,
cretere ce poate fi explicat pe baza efortului i rolului metabolic mai intens
(inclusiv la nivel hepatocitar).
Diferenele nesemnificative (p>0,05) ale activitii GPT-ului ntre cele dou
loturi cu vrste diferite, indic o relevan mai sczut a acestuia ca indicator
biochimic sanguin, inclusiv al funciei hepatocitare.
De asemenea, i fosfataza alcalin este un parametru care nu sufer
fluctuaii semnificative n funcie de vrst, la cervidee.

BIBLIOGRAFIE

1. Bentley A., 1999 An Introduction to Deer of Australia, Bunyip, Sydney


2. Dojan N., 2000 Fiziologia animalelor domestice, Printech, Bucureti
3. Dojan N., 2003 Compendiu de fiziologie animal, Printech, Bucureti
4. Fanning L., 2001 Mountain Majesty, Mill Ponn Press, USA
5. Nesterov V., 1987 Bolile vnatului, Ceres, Bucureti, 1987.

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ASPECTE CLINICE, HEMATOLOGICE I ECOGRAFICE


NREGISTRATE N UNELE AFECIUNI ALE SPLINEI
LA CINE
CLINICAL, HEMATOLOGICAL AND ULTRASONOGRAPHIC INVESTIGATIONS
IN SPLENIC DISEASES IN DOG

M.D. CODREANU, V. CRIVINEANU,


Gh. SOLCAN, Iuliana CODREANU, M. CORNIL
U.S.A.M.V. Bucureti

The present paper represents our investigations concerning the


ultrasonographic modifications of this organ corelated with the clinical and
paraclinical changes registered in dog pathology.
The investigations has been performed on 28 dogs which were
ultrasonographic examinated and included in this study.
The advantages of the echographic investigation (non-invasive and
performed in real time) and the value of the obtained results recomand this
kind of exam for certifying diagnosis, evaluating the spleen echostructure
and echogenity, in different degrees of spleen diseases.
The study is based on the data obtained in the Internal Medicine
Clinic of the Veterinary Faculty and is completed by supplementary
hematotological and biochemical tests of these patients.
This present work reveals the importance of the splenic
ultrasonography for evaluating the state of health of the dog patients. This
method of investigation is perfect integrated in the current veterinary
practice of splenic diseases diagnosis in dog.

SCOPUL LUCRRII

Scopul lucrrii de fa este de a ncerca stabilirea unor corelaii ntre


modificrile de ordin clinic, hematologic i ultrasonografic n unele afeciuni ale
splinei la cine.
Sunt prezentate rezultatele investigaiilor proprii n urma evalurii
ecografice a splinei n corelaie cu rezultatele examenului clinic i hematologic la
cinii inclui n observaiile noastre (n = 28), crora li s-a indicat efectuarea unui
examen ultrasonografic.
Particularitile speciale ale acestui organ (organ limfatic situat pe traiectul
circulaiei sangvine) i particularitile morfostrucurale l recomand drept un
organ filtru, toate afeciunile sistemice de natur n special septic influennd
parametrii de ecostrucur splenic lucru care s-a dorit a fi evideniat n prezenta
lucrare.

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Faptul c simptomatologia afeciunilor splenice este nespecific i este


corelat n foarte multe cazuri,cu afeciuni sistemice grave face ca examinarea
organului s fie de mare ajutor n corelarea datelor anamnetice, n orientarea
diagnosticului i a eventualelor examene suplimentare.
Avantajele examenului ecografic (non invaziv i realizat n timp real),
alturi de valoarea datelor obinute l recomand ca examen de elecie pentru
depistarea prin intermediul evaluarii parametrilor de ecostructur i ecogenitate
splenic, a unor afeciuni de gravitate i intensitate diferit.

LOCUL DE DESFURARE

Studiul descris n prezenta lucrare a fost realizat n cadrul Clinicii de


Medical a Facultii de Medicin Veterinar Bucureti, n perioada 2002 2005,
pe un numr de 28 de subieci canini de diverse rase i vrste, la care investigaia
ultrasonografic a splinei a constituit obiectivul prioritar sau conjunctural al
evalurii pacienilor prezentai n vederea evalurii clinice, ultrasonografice i
paraclinice i pentru stabilirea unui protocol terapeutic optim, adecvat
diagnosticului n cauz.
Investigaiile de ordin clinic i cele ultrasonografice au fost realizate n
cadrul Clinicii Medicale a Facultii de Medicin Veterinar, iar examenele
hematologice (i biochimice sanguine) au fost efectuate n cadrul Laboratorului
Clinicilor, aparinnd Facultii de Medicin Veterinar Bucureti.

MOD I METOD DE LUCRU

Investigaiile ultrasonografice au fost realizate cu ajutorul unui ecograf


Concept Dynaming Imaging (cu sond microconvex cu dubl frecven 5-7,5
MHz) i a unui ecograf PICUS - Doppler color, Pie-Medical (cu sond cu tripl
frecven 5-7,5-10 MHz), la Clinica de Medical a Facultii de Medicin
Veterinar Bucureti.
Animalele supuse examenului ecografic, au fost pregtite corespunztor
prin ndeprtarea prului de pe abdomen i au fost contenionate i poziionate n
decubit dorsal. Pe zona abdominal a fost aplicat gelul ecografic, iar dup
realizarea timpului de ateptare (10-20 minute) s-a procedat la efectuarea
examenului ultrasonografic.
Principalul obiectiv al studiului a fost aprecierea parametrilor de topografie,
volum, ecostructur i ecogenitate splenic, datele obinute prin examenul
ecografic fiind coroborate cu analize hematologice i biochimice dup caz acolo
unde acest lucru se impunea .
Din punct de vedere hematologic au fost analizai urmtorii indicatori:
numrul de eritrocite, hematocrit, numrul de elemente figurate albe,
hemoglobina, hematocritul i Volumul Eritrocitar Mediu (V.E.M.) i numrul de
trombocite.

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n unele cazuri au fost disponibil formula leucocitar i analiza calitativ a


elementelor figurate roii.

REZULTATE I DISCUII

Rezultatele investigaiilor clinice au fost menionate i nregistrate n fia


individual, ulterior au fost centralizate i sistematizate, iar aspectele ecografice
relevante sunt prezentate n cele ce urmeaz.
Pentru fiecare caz menionat datele sunt nscrise sub form de fi
individual la care a fost ataat imaginea ecografic precum i rezultatele
examenelor paraclinice sub form de tabelul 1.
Tabelul 1
Descrierea cazurilor investigate ecografic

Nr.crt. Rasa Sex Vrst


1. Pitbull Mascul 4 ani
2. Caniche pitic Mascul 7 ani
3. Bichon maltez Femel 11 ani
4. Ciobnesc german Femel 12 ani
5. Pekinez Femel 10 ani
6. Snautzer uria Mascul 8ani
7. Metis Femel 11 ani
8. Rottweiller Femel 3 ani
9. Ciobnesc german Mascul 8 ani
10. Ciobnesc mioritic Mascul 7 ani
11. Ciobnesc german Femel 6 ani
12. Metis Femel 7 ani
13. Bull Terrier Mascul 11 ani
14. Ciobnesc carpatin Mascul 4 ani
15. Golden Retriever Femel 8 ani
16. Rottweiller Femel 10 ani
17. Teckel Femel 12 ani
18. Cocker spaniol Mascul 8 ani
19. Boxer Mascul 6 ani
20. Ciobnesc german Mascul 10 ani
21. Metis Mascul 9 ani
22. Brac german Femel 9 ani
23. Ciobnesc german Femel 11 ani
24. Ciobnesc german Femel 10 ani
25. Teckel Mascul 17 ani
26. Caniche Femel 14 ani
27. Rottweiller Mascul 6 ani
28. Collie Femel 2 ani

Analiza individual i sintetic a rezultatelor investigaiilor ecografice a


evideniat faptul c splina poate prezenta modificri de topografie, volum,

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ecostructur i ecogenitate care s nu fie corelate ntotdeauna cu existena unor


modificri clinice.
Tabelul 2
Modificri clinice i ecografice nregistrate la pacienii investigai

Nr.caz. Semne clinice nregistrate Modificri ecografice


Sensibilitate la abordarea palpatorie. Distensie
1. Splenomegalie moderat
abdominal moderat.
Sindrom de febr. Anorexie, apatie, sensibilitate la
2. Abces splenic
palparea pe aria splenic.
Aprarea abdominal. Sindrom de febr. Anorexie,
3. polidipsie, sensibilitate moderat la palparea Abces splenic
abdomenului.
Hipertermie, disorexie, polidipsie, sensibilitate
4. Abces splenic.
dureroas la palparea pe aria splenic.
Sindrom de febr. Anorexie, apatie, sensibilitate
5. Abces splenic.
dureroas la palpare.
Distensie abdominal. Acumulare de lichid la nivel Formaiune
6. abdominal (ascit). La palpare se identific o macronodular
formaiune ce proemin pe suprafaa splinei. intraparenchi-matoas.
Splina se identific palpatoriu la nivelul prepubian
7. cu tendina de a invada abdomenul stng i mediu. Splenomegalie evident.
Fenomene digestive discrete.
Semne clinice terse, fr interesarea sau
8. Splenomegalie evident.
corelarea cu modificrile ecografice nregistrate.
Abordarea palpatorie a splinei este posibil la nivel
9. prepubian cu tendina de a invada abdomenul Splenomegalie evident.
posterior. Fenomene digestive discrete
10. Fr corespondent clinic Chist splenic.
Abordarea splinei poate fi realizat n plan medio-
11. lateral stg. pn la nivelul bazinului, cu evidente Splenomegalie evident.
fenomene digestive (vom, diaree).
Fr corespondent clinic. n antecedente din
12. Hematom splenic.
anamnez reiese expunerea la un accident rutier.
Semne clinice discrete, fr corelaii directe cu Splenomegalie evident.
13.
modificrile ecografice depistate. Aspect congestiv.
Distensie abdominal, distensie gastric prin
Torsiune incomplet de
14. coninut gazos, sensibilitate la palparea
splin.
abdomenului. Inapeten, vom.
Abdomen destins de lichid abundent, slbire
progresiv, anorexie. Semne de insuficien Splenomegalie moderat.
15.
cardiac (congestie pulmonare, oboseal, Ascit sever.
dispnee, tuse)
Prezena la palpare pe aria splenic a unei
16. formaiuni de dimensiuni mari, contur regulat, Splenomegalie evident.
moderat sensibilitate la abordarea palpatorie.
Slbire progresiv, inapeten, discrete modificri
17. Tumor splenic.
digestive.
Distensie abdominal, ascit moderat, vom, Formaiune tumoral
18.
inapeten, slbire progresiv. intraparenchimatoas.

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Nr.caz. Semne clinice nregistrate Modificri ecografice


Sindrom de febr, splenomegalie, anemie, icter,
19. Splenomegalie evident.
hemoglobinurie (Diagnostic confirmat: Babesioz)
Identificarea splinei se realizeaz la nivelul
Formaiune tumoral
20. abdomenului posterior (n plan medio-lateral stg.)
gigant la nivel splenic.
evidente fenomene digestive (vom, diaree).
Abdomen retractat, fr sensibilitate evident pe
21. aria de proiecie a splinei. Fr fenomene de ordin Tumor splenic.
digestiv.
Prezena la palpare pe aria splenic a unei
22. formaiuni de dimensiuni mari, sensibilitate la Tumor splenic.
palpare, discrete fenomene de staz.
Splina mult mrit n volum, margini regulate,
23. sensibilitate evident la abordarea palpatorie, Tumor splenic.
semne digestive discrete.
Sindrom febril. Anorexie, apatie, sensibilitate
24. evident la palparea pe aria splenic. Ascit, Abces splenic.
distensie abdominal evident.
Fenomene discrete, cu aprare abdominal i
25. Tumor splenic.
semne de intoleran digestiv.
Discrete semne clinice, fr interesarea sau Formaiune tumoral
26.
corelarea cu modificrile ecografice identificate. splenic.
Poliadenopatie (ln. explorabili mult mrii n volum,
Splenomegalie cu
27. fr sensibilitate). Distensie abdominal,
adenopatie evident.
sensibilitatea crescut la palpare. Anorexie, vom.
28. Fr modificri clinice. Chist splenic.

n studiul prezentat, modificrile de volum au atras invariabil i modificri


de topografie splenic. Neomogenitile splenice, difuze sau localizate, au avut n
general un corespondent clinic real, oferind indicii importante privind
diagnosticul n cauz. Modificrile astfel nregistrate, corelate cu extensivitatea i
invazivitatea procesului patologic, au permis stabilirea elementelor de prognostic.
Modificrile de volum splenic pot fi singulare (fiind vorba de entiti
morbide distincte), dar pot fi i rezultatul unei afeciuni sistemice (babesioz,
insuficien cardiac, insuficien hepatic etc.). Semnele suferinei splenice sunt
nespecifice, ele putnd uneori trece neobservate sau chiar lipsi, alteori ele fiind
violente (cum un veritabil corespondent clinic), chiar aparinnd strilor de
urgen, aa cum este cazul torsiunii gastro-splenice, nregistrat n 3,5% din
cazuri (n = 1), sau al rupturii splenice.
Modificrile ecografice n sensul alterrii ecogenitii i/sau ecostructurii
parenchimului splenic pot fi ncadrate n dou mari categorii: localizate i difuze.
n cazul congestiei splenice ecogenitatea splenic rmne perfect omogen,
dominant fiind splenomegalia evident.
Chisturile splenice, nregistrate n 7,2% din cazuri (n = 2) sunt reprezentate
de colecii lichide anecogene delimitate de un perete fin. Datorit coleciei
lichidiene (slabei ecogeniti), ele sunt nsoite de fenomenul de ntrire

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posterioar. Hematoamele splenice, n acest studiu, au nregistrat o inciden


relativ redus, de 3,5% din cazuri (n = 1), mbrcnd aspecte ecografice
caracteristice. Ecogeniatea hematoamelor depinde de momentul n care au fost
surprinse, aspectul iniial fiind hiperecogen i ulterior prezint insule anecogene.
n cazul hematoamelor mari acestea pot fi iniial hipoecogene. Abcesele splenice,
nregistrate n 17,8% din cazuri (n = 5) recunosc de asemenea un aspect
caracteristic, contur relativ regulat, coninut hipo/anecogen, perete bine
reperezentat (mediu spre nalt ecogen). Tumorile splenice primare sau secundare,
difuze sau localizate, au nregistrat 35,8% din cazuri (n = 10), fiind responsabile
de modificri de volum, de contur, de ecostructur i ecogenitate la nivelul
parenchimului splenic, cu localizare i ntindere diferit.
Examenele paraclinice au venit n completarea examenului ecografic,
dominante fiind investigaiile hematologice i biochimice. De multe ori, n cazul
tumorilor splenice, s-a apelat la splenectomie pentru punerea diagnosticului de
certitudine.

CONCLUZII

1. Studiul de fa a fost efectuat pe 28 de cazuri, considerate relevante, cu


modificri ecografice ale splinei. Examenul ecografic a permis evaluarea
topografiei, dimensiunilor (volumului), ecostructurii i ecogenitii parenchimului
splenic la cine, iar evaluarea ecografic s-a realizat n strns corelaie cu
rezultatul investigaiilor clinice i cu eventualele modificri paraclinice i
ecografice conexe.
2. Modificrile de topografie au fost induse de torsiunea, congestia i
splenomegalia (cu etiologie diferit) difuz i/sau focalizat.
3. Modificrile de volum (focalizate sau difuze) au avut drept corespondent
entiti morbide distincte i specifice, pentru fiecare dintre acestea.
4. Modificrile de ecostructur splenic (leziuni focale) au fost reprezentate
de abcese, n 17,8% din cazuri (n = 5), hematoame, n 3,5% din cazuri (n = 1),
chisturi, n 7,2% din cazuri (n = 2) i neomogenitile focale ce nsoeau
afeciunile neoplazice (primare sau metastazice).
5. Splenomegalia nregistrat n 32% din cazuri (n = 9) a recunoscut
diferite cauze (insuficien cardiac, babesioz, limfom/limfosarcom).
6. Babesioza din punct de vedere ecografic a fost nsoit constant de
splenomegalie, iar clinic de anemie, icter, hemoglobinurie i hemoglobinemie.
7. n categoria afeciunilor neoplazice, nregistrate n 35,8 % (n = 10),
tumorile difuze ale splinei au fost generate preponderent de afeciuni cu origine
limfatic i sangvin (limfoame i limfosarcoame), suferind constant modificri
de volum i ecogenitate i inconstant de ecostructur, care coroborate cu
rezultatele examenului hematologic certific diagnosticul. Din punct de vedere
ecografic, hemangiosarcoamele s-au caracterizat prin heterogenitate crescut, cu

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existena constant a cavitilor hipo/anecogene mrginite de zone nalt ecogene.


Analiza comparativ a rezultatelor investigaiilor ecografice n cazul tumorilor
altor organe abdominale a evideniat faptul c splina este una dintre organele care
pot dezvolta tumori de dimensiuni mari sau foarte mari (gigant).
8. Simptomatologia n cazul afeciunilor splenice investigate a fost
nespecific, incluznd i semne la distan care in de afectarea sistemic i a
organelor de la nivelul cavitii abdominale.
9. Se poate aprecia c prin coroborarea rezultatelor investigaiilor
ecografice (ofer date privind topografia, volumul, ecostructura, ecogenitatea
organului) cu cele ale examenului clinic, hematologic i biochimic se poate stabili
diagnosticul de certitudine.

BIBLIOGRAFIE

1. Badea, I.R., Dudea, S.M., Mircea, P.A., Stamatian, Fl., 2000 Tratat de
ultrasonografie clinic, vol.I; Ed. Medical, Bucureti.
2. Bhadwal, M.S., Mirakhur, K.K., 2001 - Sonography of the canine healthy and
diseased spleen, Veterinary Bulletin, vol.71, No. 3, Art. 2103, Pag. 339.
3. Burk R.L., Ackerman N., 1996 - Small Animal Radiology and Ultrasonography. A
nd
diagnostic atlas and text, 2 Edition; W.B. Saunders Company, U.S.A.
4. Codreanu, M., 2000 - Aspecte ecografice nregistrate n cazul tumorilor abdominale
la cine; Al VIII-lea Congres Naional de Medicin Veterinar, Bile Felix, 17-20
octombrie.
5. Codreanu, M., 2001 - Importana examenului ecografic n diagnosticul coleciilor
cavitare la carnivore.; Revista Romn de Medicin Veterinar, vol. 11, nr.2,
p.139-146.
6. Codreanu M, Diaconescu A., 2003 Diagnosticul ecografic la animalele de
companie, Ed. Coral Sanivet, Bucureti.
7.
Crnganu, D., Codreanu, M., Preda, Cristina, Mocanu, Jaqueline, Britreanu,
S., 2001 - Diagnosticul oncologic multidisciplinar i diferenial ntre leucemiile
limfoide i limfom la cine, Simpozion a 140 ani Alma Mater Veterinaria
Bucurescensis, Ed. All, pg. 84, Bucureti, 2001.
8. Ramirez, S., Douglass, J.P, Robertson, I.D., 2002 - Ultrasonographic features of
canine abdominal malignant histiocytosis, Veterinary Bulletin, vol. 72, No. 8, Art.
6248, Pag. 1099.

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REZULTATELE INVESTIGAIILOR CLINICE I


ULTRASONOGRAFICE N DIAGNOSTICUL
REVRSATELOR CAVITARE LA CINE I PISIC
CLINICAL AND ULTRASONOGRAPHIC INVESTIGATIONS
IN CAVITARY EFUSIONS IN DOGS AND CATS

M.D. CODREANU
U.S.A.M.V. Bucureti

Clinical and interventional ultrasonography is considered the


prefferable method of investigation of the parenchimatous and/or cavitary
organs in veterinary practice, for the non-invasive character and for its
valuable real time results.
The utility of the ultrasonographic investigation in parenchimatous
organs diagnosis is well known. Althought, based on the physical properties
of the intraparenchimatous effusions (abcesses, hematoms, chists) and
intracavitary effusions (around the abdominal organs), these liquid
acumulations are easy to identify on the ultrasonographic image. The
ultrasonographic images of the effusions are characterised as omogene
anechoic zones through which the ultrasonic echoes pass whithout
resistence.
The present paper aim to reveal the results of our work in presenting
the importance and the limitations of ultrasonographic exam in diagnosis of
the cavitary effusions in dogs and cats.

SCOPUL LUCRRII

n lucrarea de fa am ncercat s conturm tabloul clinic propriu


acumulrilor de lichid la nivelul diferitelor caviti ale organismului n strns
corelaie cu aspectul ecografic pentru fiecare din localizri, pentru a evidenia
relevana acestui examen de imagine n diagnosticul complex al revrsatelor
cavitare la animalele de companie.
Prezentarea cazurilor este realizat pe categorii de afeciuni, respectiv n
funcie de sediul acumulrii de lichid, imaginile ecografice relevante fiind
prezentate i descrise n cadrul lucrrii, iar dominantele clinice sunt grupate de
asemenea n funcie de spaiul, cantitatea i tipul revrsatului.

LOCUL DE DESFURARE

Examenele clinice i utrasonografice au fost realizate pe 36 cini i pisici


cu diverse afeciuni, care prezentau efuziuni cavitare, la nivelul spaiului

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peritoneal, pleural sau pericardic, sau policavitare prezentate n vederea evalurii


la Clinica de Medical a Facultii de Medicin Veterinar Bucureti.

MOD I METOD DE LUCRU

Investigaiile ultrasonografice la animalele cu diferite acumulri cavitare


(suspicionate pe baza datelor clinice i ale altor tehnici de investigaie paraclinic)
s-au realizat n perioada 2002 2005 la Clinica Medical a Facultii de Medicin
Veterinar Bucureti, cu ajutorul : unui ecograf Concept Dynamic Imaging,
folosind o sond sectorial microconvex cu dubl frecven 5 - 6,5 MHz i a unui
ecograf Picus Pie Medical cu sond convex cu tripl frecven: 5; 7,5 i
respectiv 10 MHz.
Acumulrile de lichid de la nivelul cavitii abdominale, toracice i a
spaiului pericardic pot fi identificate i accidental la examenul ecografic de rutin,
prin vizualizarea unei colecii de lichid situate post -, sau prediafragmatic.
n cazul acumulrilor de la nivelul cavitii peritoneale, acestea sunt
identificate cu uurin datorit aspectului anecogen, situat ventral de sonda
ecografic.
Examenul general al abdomenului, premergtor investigrii diferitelor
organe abdominale, se realizeaz pentru obinerea unei imagini de ansamblu a
peretelui abdominal i a topografiei organelor abdominale.
Cauzele revrsatelor cavitare sunt extrem de numeroase, insuficiena
cardiac dreapt, hipoproteinemia, neoplasmele, infeciile specifice, leziuni
posttraumatice soldate cu compresiuni vasculare, rupturi vasculare (hemotorax)
sau ale vaselor limfatice (chilotorax), procese inflamatorii etc.
Natura coleciei lichide este greu de apreciat ecografic, dar existena unor
cauze evidente, ca i asocierea unor entiti morbide de aceeai natur (afeciuni
ale cordului drept, prezena unor formaiuni la nivelul pericardului, pleurei,
torsiunea unui lob pulmonar sau hernia diafragmatic), inclusiv alte epanamente
cavitare (pericardice, peritoneale), pot susine un anumit diagnostic etiologic
prezumptiv.
n cazul suspiciunii acumulrilor de lichide de la nivel cavitar toracice s-a
procedat la tunderea/raderea zonelor cutanate corespunztoare (regiunea
abdominal ventral, spaii intercostale, zona abdominal lateral), care au servit
ca fereastr acustic pentru investigarea zonelor de interes i s-a aplicat gelul
ecografic, dup care s-a trecut le efectuarea examenului ecografic.

REZULTATE I DISCUII

Cazurile prezentate n vederea evalurii la Clinica de Medical a Facultii


de Medicin Veterinar Bucureti au fost examinate clinic i utrasonografic,
realiznd fie individuale, grupate n funcie de sediul i tipul afeciunii, pe cini
i pisici care prezentau efuziuni cavitare, la nivelul spaiului peritoneal, pleural
sau pericardic, sau acumulri policavitare.

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n lucrarea de fa descrierea diferitelor cazuri de cini i pisici este fcut


n funcie de categoria de afeciuni, respectiv n funcie de sediul acumulrii de
lichid. Rezultatele examenului ecografic au fost consemnate n fiele individuale,
iar imaginile cele mai relevante sunt prezentate n cele ce urmeaz, iar semnele
clinice nregistrate au fost grupate de sediul acumulrii, cantitatea i tipul acesteia.
Aprecierea naturii coleciei lichide doar pe baza examenului ecografic a
fost dificil, dar existena asociat a unor entiti morbide, cum ar fi (afeciuni ale
cordului drept, prezena unor formaiuni la nivelul pericardului, pleurei, decelarea
unor formaiuni nlocuitoare de spaiu la nivel parenchimatos sau cavitar,
coexistena altor epanamente cavitare au venit n completarea diagnosticului
prezumtiv.
Cazurile investigate sunt reprezentate de cini i pisici aparinnd unor rase
diferite, de vrste diferite cu afeciuni primare sau secundare de hidropizie
(tabelul 1).
Tabelul 1
Distribuia cazurilor investigate

Nr.crt. SPECIA RASA SEX VRSTA


1 Cine Metis Femel 8 ani
2 Cine Cocker spaniol Femel 10 ani
3 Cine Labrador Femel 4 ani
4 Cine Cocker spaniol Femel 8 ani
5 Cine Metis Femel 9 ani
6 Cine Metis Terrier Femel 5 ani
7 Cine Ciobnesc german Mascul 11 ani
8 Cine Setter irlandez Femel 12 ani
9 Cine Cocker american Mascul 7 ani
10 Cine Ciobnesc carpatin Mascul 6 luni
11 Cine Snautzer uria Mascul 2 luni
12 Cine Brac german Femel 9 ani
13 Cine Doberman Mascul 5 ani
14 Cine Ciobnesc german Mascul 5 luni
15 Cine Teckel Femel 16 ani
16 Cine Bichon maltez Mascul 9 ani
17 Cine Ciobnesc mioritic Mascul 4 ani
18 Cine Ciobnesc german Mascul 6 ani
19 Cine Ciobnesc german Femel 1,5 ani
20 Cine Golden Retriever Mascul 3 ani
21 Cine Doberman Mascul 7 ani
22 Cine Setter irlandez Femel 11 ani
23 Cine Caniche Mascul 14 ani
24 Cine Chow chow Femel 9 ani
25 Cine Ciobnesc german Femel 9 ani
26 Cine Ciobnesc german Mascul 7 ani
27 Pisic Persan Femel 3 ani
28 Pisic Birmanez Mascul 7 ani

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Nr.crt. SPECIA RASA SEX VRSTA


29 Cine Setter irlandez Mascul 8 ani
30 Cine Metis Femel 6 ani
31 Pisic Metis Mascul 10 ani
32 Pisic Birmanez Mascul 2 ani
33 Cine Metis Mascul 9 ani
34 Pisic Metis Femel 4 ani
35 Cine Bichon Frise Mascul 9 luni
36 Cine Caniche Mascul 14 ani

Lichidul ascitic a putut fi evideniat cu uurin prin tehnica ecografic,


graie particularitilor sale ecografice unice. Prezena n cantitate redus a
lichidelor n cavitatea abdominal, a facilitat vizualizarea organelor abdominale,
iar acumularea masiv a fluidelor reduce n primul rnd, posibilitatea abordrii
organelor de la nivelul cavitii abdominale (prin reducerea compresibilitii
peretelui abdominal).
n realizarea examenului ecografic, pentru organele cavitii abdominale, se
recomand nceperea cu abordarea celor situate n abdomenul superior i
continuarea cu investigarea celor din abdomenul posterior stng i apoi drept. La
examenul ecografic de rutin au putut fi identificate colecii de lichid att
postdiafragmatic ct i prediafragmatic (lichidotorax).
Se poate aprecia c din punct de vedere ultrasonografic structurile tisulare
de la nivelul cavitii toracice (mai puin cordul) nu au caracteristici ecografice
specifice. Evidenierea modificrilor din cavitatea toracic, prin examen
ecografic, este facil n special n cazul localizrii la nivel superficial, imediat
prediafragmatic sau n cazul acumulrilor de lichid (inflamator sau neinflamator).
Prezena epanamentelor la acest nivel se poate identifica cu uurin, sub forma
unor zone anecogene, situate ntre pulmoni i peretele toracic.
Examenul cordului - ecocardiografia s-a realizat folosind tehnici aparte de
abordare, msurare i interpretare, apelnd la: modulul B pentru aprecierea
dimensiunilor, ecogenitii i micrilor diferitelor structuri i modulul M care
permite realizarea reprezentrii grafice a unei poriuni izolate a cordului i
micrile acestuia n raport cu alte formaiuni anatomice, precum i msurarea
diametrelor cavitilor cordului.
Am optat pentru cele trei abordri ecografice recomandate pentru ecografia
cordului: fereastra parasternal dreapt (la nivelul spaiilor intercostale IV i V,
ntre stern i jonciunea costocondral), fereastra parasternal stng caudal
(apical): la nivelul spaiilor intercostale V-VII n imediata vecintate a sternului
i respectiv fereastra parasternal stng cranial: la nivelul spaiilor intercostale
III IV, ntre stern i jonciunea condrocostal. Ca i n medicina uman imagini
satisfctoare am putut obine prin plasarea sondei imediat napoia apendicelui
xifoidian.

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Existena asociat a unor entiti morbide: afeciuni ale cordului drept,


prezena unor formaiuni la nivelul pericardului, pleurei, torsiunea unui lob
pulmonar sau hernia diafragmatic, sau alte epanamente cavitare (pericardice,
peritoneale) vin n completarea diagnosticului prezumtiv. Obinuit toracocenteza
i respectiv analiza lichidului de puncie sunt relevante (examen citologic).
Existena asociat la nivelul cavitilor de multe ori, corelat cu
simptomatologia i cu examenul lichidului de puncie reprezint suficiente
elemente, care pot conduce la un diagnostic de certitudine.
Sunt frecvente situaii n care datele anamnetice sunt extrem de sugestive n
ceea privete etiologia epanamentului cavitar i tipul acesteia (ex. traumatismele,
accidente rutiere etc.)
Afeciunile diafragmului au putut fi puse n eviden cu ajutorul tehnicii
ecografice att cele de la nivelul feei anterioare, ct i cele de la nivelul feei
posterioare.
Herniile diafragmatice sunt uor de evideniat, de exemplu hernierea unui
lob hepatic sau hernierea unui lob pulmonar (caudal sau accesoriu).
Cel mai frecvent este implicat i antrenat ficatul n hernierea
transdiafragmatic, caz n care, prezena unei structuri anterior diafragmului (dar
cu structura i ecogenitatea caracteristic ficatului) certific diagnosticul.

CONCLUZII

1. Acumulrile de lichide de la nivelul cavitii toracice pot fi identificate


folosind drept fereastr acustic spaiile intercostale sau prin abordarea la nivel
abdominal (transdiafragmatic).
2. Acumulrile de la nivelul cavitii peritoneale pot fi identificate cu
uurin datorit aspectului anecogen (caracteristic coleciilor de lichid), situate
ventral de sonda ecografic.
3. Tehnica ecografic este mai precis n evidenierea fluidelor peritoneale,
comparativ cu tehnica radiografic.
4. Natura acumulrilor de lichid nu poate fi precizat cu ajutorul tehnicii
ecografice, dar poate fi evideniat existena celularitii crescute i/sau
acumulrile de fibrin.
5. Examenul ecografic este util n realizarea punciilor cavitare (att n scop
de diagnostic, ct i aspirativ-evacuator), fiind recomandat n special pentru
stabilirea omogenitii lichidului, ct i pentru evitarea penetrrii unor eventuale
structuri aderente peretelui abdominal intern (n regiunea propus pentru
realizarea centezei).
6. Cu limitele amintite (generate de imposibilitatea abordrii
corespunztoare a structurilor de interes), examenul ecografic furnizeaz elemente
de diagnostic extrem de importante, plasnd tehnica ultrasonografic n ansamblul
de diagnostic imagistic al afeciunilor cavitii toracice.

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7. Caracteristicile ecografice generale ale mediilor lichide, relevana i


acurateea rezultatelor obinute, recomand examenul ecografic ca o veritabil
alternativ de diagnostic, de un real folos n abordarea clinic i terapeutic a
efuziunilor cavitare.

BIBLIOGRAFIE

1. Burk R.L., Ackerman N., 1996 - Small Animal Radiology and Ultrasonography. A
diagnostic atlas and text, 2nd Edition; W.B. Saunders Company, U.S.A.
2. Codreanu M., 2000 - Examenul ecografic n diagnosticul epanamentelor cavitare la
carnivore. Al VIII-lea Congres Naional de Medicin Veterinar, Bile Felix, 17-20
Octombrie.
3. Codreanu M., 2001 - Importana examenului ecografic n diagnosticul coleciilor
cavitare la carnivore. Revista Romn de Medicin Veterinar (ISSN 1220-
3173), vol. 11, nr.2, p.139-146.
4. Codreanu M, Diaconescu A., 2003 Diagnosticul ecografic la animalele de
companie, Ed. Coral Sanivet, Bucureti.
5. Han C.M., Hurd C.D., Kurklis L., 1994 - Practical Guide to Diagnostic
Imaging:Radiograpy and Ultrasonography; American Veterinary Publications,
Inc.
6. Ionacu I., Codreanu M., J. Mocanu, 2002 Chilotoraxul la pisic prezentare
caz - Sesiune tiinific dedicat srbtoririi a 150 de ani de la nfiinarea
nvmntului agricol n Romnia, 11 octombrie, FMV. Bucureti.
7. Lungu A., Codreanu M., Balint E., Constantinescu C., Brslau D., Fernoag C.,
2003 Dubl localizare la nivelul seroaselor cavitii toracice la pisic. Al XXXIV-
lea Simpozion Naional de Morfologie Normal i Patologic Actualiti n
patologia aparatului cardiovascular, Institutul Naional Victor Babe, 5-7
Noiembrie, pag. 58-59.
8. Penninck D.G., 1998 - Ultrasonography; Veterinary Clinics of North America, Small
Animal Practice, 28(4).
9. Pop T., 1998 - Ecografie clinic diagnostic i intervenional; Editura Medical,
Bucureti.

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MODIFICRI HEMATOLOGICE, BICHIMICE SANGUINE


I HISTOPATOLOGICE INDUSE DE SULFATUL DE
CUPRU LA OAIE
HEMATOLOGICAL, SERUM BIOCHEMICAL AND HYSTOPATHOLOGYCAL
CHANGES INDUCED BY COPPER SULPHATE IN SHEEP

G.V. GORAN, M.D. CODREANU,


V.CRIVINEANU
U.S.A.M.V. Bucureti

The investigations described in this study were realised on adult


sheep batches, after the oral administration of CuSO4 , in different doses,
from 5 to 20 mg/kg body weight (0,1% solution), depending on the
experimental batch, after this were realised the laboratory analysis.
The hematological investigations reveals an obvious
hemoconcentration (the elevation of the RBC and of the PCV), in
correlation with the dehydration consecutively status (induced by the
polyuria and diarrhoea), esspecially in ewes from the batches were was
administered the cooper sulphate in doses of 10 and respectively 20 mg/kg.
From the biochemical point of view, consistent elevations registered
the seric specific hepatic enzymes (esspecially ALAT), at ranges of 92 UI
and 118 UI (for last two doses) changes what indicate the hepatic
alterations and of the creatininemia and uremia, what also indicate and
obvious nephrotoxicity (esspecially in doses of 20 mg/kg).
The most frequent lesional aspects were the congestive and the
hemoragic lesions identified in liver, kidneys and gastrointestinal tractus,
in the same time the cytological examinations reveals an intensive
hemolysis, hepatic cell distructions and granulo-vacuolary dystrophy,
distructions of the renal epithelium. From hystopathologycal point of view,
the most frequent aspects were the evident mesenchymal reactions in
portobiliar space in the liver, and in kidney more relevant were the
congestions in renal parenchym and the tubular cell distructions (with the
remaining of the basal membrane), intraglomerular hypercellularity. In the
same time, constant myocardic lesions were the congestions and the
mesenchymal reactions correlated with microvacuolisations in miocardyc
cells.
All this results reveals the intensivity and the extensivity of the
pathological processes, correlated with the administration of the copper
sulphate at this doses in sheep.

Copper sulphate is frequently involved in inducing of accidental poisoning


in animals. The ruminants, especially the sheep, are more sensistive to CuSO4
intoxications.

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Compounds containing this salt are steel in fitosanitary and


phramacotherapeutic use, and consequently the risk of oral intake of large
amounts or supradosing in animals, is continuously a realy problem in veterinary
medicine.
The purpose of this study is achieving important informations about the the
hematological and biochemical parameters variations, and histopathologycal
changes, after administration of coppers ulphate in different doses in sheep, and to
establish possible correlations between the degree of the registered changes and
the used dose.

MATERIAL AND METHODS

The experimental investigations were realised on adult sheep batches


(n=6), after oral administration of CuSO4 solution 1%o, in doses ranging from 5 to
20 mg/kg b.w. The experiment wass carried out on 4 batches, the first (control
batch) and 3 experimental batches in which we used the individual dose of 5, 10
and respectively 20 mg/kg b.w.
After the administration (24 hours), were taken blood samples for
laboratory analysis.
The hematological investigated parameteres were the levels of direct
values erythrocytic constants: Red Blood Cells (RBCs = mil/l), hemoglobinemia
(Hb = g/dl), Hematocrit = Packed Cell Volume (PCV = %) and the values of some
derived erythrocytic parameters: Mean Corpuscular Volume (MCV = 3), Mean
Corpuscular Hemoglobin (MCH = pg), Mean Corpuscular Hemoglobin
Concentration (MCHC = g/dl).
The biochemical investigation weas realised on blood serum, and was
consistiing from the determination of seric transaminase activity (ALAT and
ASAT), creatininemia and uremia.
After the sacrification, the lesional changes were revealed by nechropsic
and histopathologyc examination (using the Hematoxiline-Eosine-Methilen blue
method).

RESULTS

The results of the hematologycal investigations of the experimental batches


in comparison with the results obtained in control batch are presented in table 1.
The hematological investigations reveals an obvious hemococncentration
(the elevation of the RBC and of the PCV) in correlation with the dehydration
consecutively status (induced by the the polyuria and diarrhoea), esspecially in
ewes from the the batches were was administered the copper sulphate in doses of
10 and respectively 20 mg/kg b.w.
The results of the biochemical investigations are presented in table 2.

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Table 1
Average values of the hematological parameteres
in control and in experimental baches

Batch/ Control Batch No.1 Batch No.2 Batch No.3


Parameters batch (5 mg/kg b.w.) (10 mg/kg b.w.) (20 mg/kg b.w.)
RBCs 9.23 9.64 10.92 11.56
Hb 7.3 7.76 8.3 9.26
Hct 27.55 29.15 34.03 33.7
MCV 30.11 30.29 31.3 29.2
MCH 7.93 8.05 7.85 8.07
MCHC 26.43 26.73 24.14 27.67

Table 2
Average values of the biochemical investigated parameteres

Batch/ ASAT ALAT Uremia Creatininemia


Parameters (UI) (UI) (mg/dl) (mg/dl)
Control batch 69.16 7.75 35.16 1.45
Batch No.1
69.83 6.7 35.16 1.40
(5 mg/kg b.w.)
Batch No.2
92.66 9.5 72 1,65
(10 mg/kg b.w.)
Batch No.3
118.3 13.5 95.16 2.14
(20 mg/kg b.w.)

From the biochemical point of view, cosistent elevations registered the


seric specific hepatic enzymes activity (esspecially ASAT at ranges of 92.66 UI
and 118 UI, for last two doses), changes what indicates the hepatic alterations,
and of the uremia and creatininemia, which also indicate and obvious
nephrotoxicity (esspecially in doses of 20 mg/kg b.w.).
The morphopathologycal changes are very severe, esspecially in case of
using the dose of 20 mg copper sulphate/kg b.w. Generally, the most frequent
changes were congestive and hemoragic lesions in gastrointestinal tractus and
parenchymatous organs.
In liver, from the hystopathologycal point of view, more constant and
consistent changes were the congestive and hemoragic aspects, correlated with
hepatic cell distructions, evident mesenchymal reactions in portobiliary space.
In kidneys, more relevant changes were the congestions in renal parenchym
corticaly and medulary areas, intravasculary and intraglomerulary hemolysis
and congestion in glomerular spaces and intraglomerular hypercellularity, and
tubular cell distruction with the remaining of the basal membrane.
In pulmonary and myocardic tissues samples, the lesions were less severe
and inconsistent.
These lesions were more severe and constant in animals from the batch
were was used the copper sulphate in 20 mg/kg b.w. dose.

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CONCLUSION

The administration of CuSO4 in sheep, at doses beginning from 5 mg/kg


b.w. to 20 mg/kg b.w., was followed by consistent changes of the hematological
and biochemical parameteres, correlated with the specific induced action at this
level.
The hematological investigations reveals an constant hemoconcentration,
due to the consecutively dehidration and an evident hemolysis.
The biochemical changes were dominated by an consistent elevation of the
seric enzymatic activity (especially ASAT), which indicate serious hepatic cell
morphologic and functional alterations, and high values of the uremia and
creatininemia, which also indicate the nephrotxicity of this salt in sheep.
The consistant and constant histopathologycal aspects can represent a realy
morphologycal support of these paraclinical modifications, esspecially in liver
and in kidneys, induced by the copper sulphate.
The morphopathologycal registerted changes are demonstrating the acute
and brutal action of the copper sulphate for the investigated parenchymatous
organs, which indicate her extreme hepato- and nephrotoxicity.
All these results reveals the intensivity and the extensivity of the
pathological processes, correlated with the oral administration of the copper
sulphate (at these doses), in sheep.

REFERENCES

1. Codreanu, M., A. Ionescu, ueanu,E., Adriana Oranu, Iuliana Codreanu, 1999


Hematological, biochemical and toxicologycal aspects in experimental
poisoning with copper sulphate in sheep; 9th Sci.Sess., Spiru Haret F.M.V.,
Bucharest, p.51-52, April, 29.
2. Codreanu, M.D., 1999 Contribuii la farmacotoxicologica unor substane
methemoglobinizante implicate n patologia ovinelor, Tez de doctorat,
Bucureti, 2 iulie.
3. Crivineanu, V., M. Codreanu, 1998 Laboratory diagnosis in veterinary toxicology,
U..A.M.V., Bucureti.
4. Crivineanu V., Goran G.V., 2004 Toxicologie veterinar, Editura Printech,
Bucureti
5. Goran G.V., Crivineanu V., Manuella Militaru, Camelia Papuc, 2004 Aspecte
enzimatice i histopatologice n intoxicaia experimental cu sulfat de cupru la
oaie, Lucrri tiinifice Medicin Veterinar, vol. XXXVII, 2004, Timioara, ISSN
1221-5295
6. Osweiller, G.D. Toxicology, The National Veterinary Medical Series for
Independent Study, Williams and Wilkins, 93, Iowa, US

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MODIFICRI HISTOPATOLOGICE I
ELECTRONOMICROSCOPICE INDUSE DE NITRATUL
DE SODIU N EXPERIMENT ACUT I CRONIC LA OAIE
HYSTOPATOLOGYCAL AND ELECTRONOMICROSCOPIC CHANGES
IN ACUTE AND CHRONIC EXPERIMENT INDUCED BY SODIUM NITRATE IN SHEEP

G.V. GORAN, M.D. CODREANU,


V.CRIVINEANU, Manuella MILITARU,
Iuliana CODREANU, Emilia CIOBOTARU
U.S.A.M.V. Bucureti

Structural changes alterations can be better observed and


understood in their deeply intimity, by understanding the ultrastuctural
(electronomicroscopical) correlated with hystopathologycal changes, in
different tisular structures after sodium nitrite administration in sheep.
For this purpose, sodium nitrite was administered in dose of 150
mg/kg (acute experiment) and 20 mg/kg (in chronic experiment) in adult
sheeps. After sacrification, were colected samples for the
anatomopathological and electronomicroscopycal examinations (TEM).
Myocardic lesions were dominated by vacuolar dystrophy and other
alterative lesions, such as congestive and necrotic modifications, and the
ultrustructural changes were dominated by intra- and interfascicular
oedema, mytochondrial vacuolisations and mytochondrial distructions,
necrosis and disruptions in myocardial fibres, and alterations in Z
myofibrilar bands .
In the liver, the lesional aspects were dominated by the vacuolar
intracytoplasmic dystrophy, caryolysis in connection with a reduced
cellularity and oedema in Disse spaces, and the ultrustuctural examinations
reveals mytochondrial distructions and vacuolisations.
In kidneys, the most important were the evident alterations of the
brush border in tubular structures, and an obvious intraglomerular
hypercellularity, with the growing of the vascular glomerulus, constant
associated with lesions of intracytoplasmic mycrovacuolisations of the
nephrocytes.

In ruminants, nitrogenous compounds intake, especially the sodium nitrite,


in high doses (in case of intoxication), is frequently involved in inducing serious
damages to different tisular structures. Morphologycal alterations can be better
observed and understood in their deeply intimity, by understanding the
ultrastuctural (electronomicroscopical) correlated with hystopathologycal
changes, in different tisular structures after sodium nitrite administration in sheep.

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MATERIAL AND METHODS

For this purpose, sodium nitrite was administered in dose of 150 mg/kg
(acute experiment) and 20 mg/kg (in chronic experiment) in 5 adult
sheeps/experiment. In acute experiment sodium nitrite was oral administered, in
1% solution. In chronic experiment sodium nitrite was oral administered in daily
dose of 20 mg/kg, for 14 days consecutively After slaughtering, were colected
samples for the anatomopathological and electronomicroscopycal examinations
(TEM). For the hystopathologic examinations were used the trichromic strains
(Hematoxiline-Eosine - Methilen blue) and Toluidine blue method.
Electronomycroscopical examinations were realised by TEM (transmision)
method.

RESULTS AND DISCUSION

The results of our study showed that the registered lesions are very
complex, with a high degree of specificity in parenchyma.
Myocardic lesions were dominated by vacuolar dystrophy and other
alterative changes, such as congestive and necrotic modifications, and the
ultrastructural changes were dominated by intra- and interfascicular oedema,
mytochondrial vacuolisations and distructions, necrosis and disruptions in
myocardial fibres, alterations in Z myofibrilar bands.
In kidneys, the most constant and consistent changes were the severe
alterations of the brush border in tubular structures, with majore reducing of the
filtration area, and an obvious intraglomerular hiperecellularity, with the growing
of the vascular glomerulus, constant associated with lesions of intracytoplasmic
microvacuolisation of the nephrocytes.
In the liver, the lesional aspects were dominated by the vacuolar
intracytoplasmic dystrophy, caryolysis, in connection with a reduced cellularity
and oedema in Disse spaces and the electronomicroscopical examinations reveals
major mytochondrial distructions and vacuolisations.
In chronic experiment, the lesional complex was evident reduced and
nonspecific, generally being dominated by moderate congestive and distrophyc
lesions in parenchymatous organs.

CONCLUSION

The given results showed the consistent changes in different tissues


(internal organs) after hystopatologycal and ultrastructural (electronomicroscopic)
examinations, in acute experiment, after sodium nitrite administration in sheep.

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These findings reveals a constant and consistent complex of changes,


registered in parenchymatous organs, and recognize a high degree of specificity
(characteristic for the acute agresion of these structures).
The lesional complex is in accordance with the registered clinical
manifestations, and with other studies, described by the specific literature.
In chronic experiment the anatomoclinical complex is less severe and
without obvious specificity.
Our results try to complete the avalanche of bybliographycal
informations about the serious damages induced by oral intake of different
amounts of sodium nitrite in sheep.

REFERENCES

1. Baba, A.I., 1997 Diagnostic necropsic veterinar, Ed. Ceres, Bucureti.


2. Codreanu M.D., Manolescu N., Adriana Oranu, Iuliana Codreanu, 2001 -
th
Ultrastructural changes induced by sodium nitrite in sheep. 5 ESVCN
th th
Conference, 13 14 September, Sursee, Switzerland.
3. Codreanu M.D., 1999 Contribuii la farmacotoxicologia unor substane
methemoglobinizante implicate n patologia ovinelor, Tez de doctorat,
Bucureti, 2 iulie.
4. Crivineanu V., Goran G.V., 2004 Toxicologie veterinar, Editura Printech,
Bucureti
5. Goran G.V., Crivineanu V., M.D. Codreanu, 2005 - A study about induced
methaemoglobinaemia by copper based products in sheep, Buletinul USAMV
Cluj-Napoca, Seria Medicin Veterinar, volumul 62, ISSN 1454-2382
6. Cornil, N., Manolescu, N., 1995 Structura i ultrastuctura organelor la animalele
domestice, Editura Ceres

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STUDIU COMPARATIV PRIVIND ARTROGRAFIA


CU SUBSTANE DE CONTRAST IONICE I NONIONICE
(ODISTON I ULTRAVIST 300)
COMPARATIVE STUDIES IN ARTROGRAPHY WITH IONIC AND NONIONIC
CONTRAST SUBSTANCES (ODISTON AND ULTRAVIST 300)

I. PAPUC, R. LCTU
U.S.A.M.V. Cluj-Napoca

The objectives of this studies is to establish the possibilities for


atrography examination utilizing Odiston, witch is a ionic contrast
substance, compare to Ultravist 300, a nonionic contrast substance, regard
the qualities of artrographyc images, optimal time for radiological
exposure, the quantities of using substance, the concentration and the side
effects witch can occur at administration.

MATERIALE I METOD

Pentru efectuarea acestui studiu am folosit 8 cini de vrst i talie diferit.


6 dintre aceti cini au fost folosii pentru determinarea dozei utile pentru
articulaiile n studiu.
Animalele au fost lotizate dup talie, astfel:
- Animale de talie mare cu greutate cuprins ntre 20-24 kg;
- Animale de talie mijlocie cu greutate medie de 10 kg.
Substanele folosite au fost reprezentate de Odiston substan de
contrast organoiodat i Ultravist 300 substan de contrast nonionic, cu
osmolalitate sczut.
Odistonul a fost foosit n concentraie de 30% ca atare, ct i diluat cu ap
distilat n raport de 1:1 i 1:3, obinndu-se soluii de 15% i 7,5% Odiston.
Substana nonionic Ultravist 300, gsit n flacoane de 100 ml cu 0,623 g
iopromid/ml n concentraie de 30%, a fost folosit ca atare, ct i diluat cu ap
distilat n raport de 1:1.
Dozele folosite au fost n funcie de mrimea articulaiei i talia animalului.
Pentru articulaia umrului doza a fost cuprins ntre 2,0-2,5 ml; articulaia cotului
2,0 ml; articulaia grasetului 2,5-3 ml, la animalele de talie mare. Pentru animalele
de talie mijlocie am folosit pentru articulaia umrului 1,5 ml; articulaia cotului 1,5
ml; articulaia grasetului pn la 2,0 ml.
Pregtirea animalului am efectuat-o prin anestezie general folosind
Acepromazin n doz de 0,15 ml/10 kg corp i.v., n asociaie cu Ketamin 10
mg/kg corp. Local s-a efectuat toaleta mecanic respectndu-se regulile de
asepsie i antisepsie.
Punciile articulare, au fost efectuate cu ace sterile, adaptate dup talia
animalului. Toate examinrile s-au efectuat folosindu-se procedura stabilit fr
premedicaie.

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Radiografiile diferitelor articulaii cuprinse n studiu au fost examinate n


funcie de intensitatea de contrast, claritatea cartilajului i a conturului sinovial.
Pentru articulaia umrului animalul a fost contenionat n decubit lateral,
cu membrul anterior liber, tras, n extensie. Puncia a fost realizat prin orientarea
acului uor oblic, pe partea lateral a articulaiei.
Pentru articulaia cotului animalul a fost contenionat n decubit lateral,
membrul uor tracionat, puncia efectund-o n faa olecranului, la aproximativ 1
cm, pn n momentul ajungerii n spaiul articular.
Pentru articulaia grasetului animalul a fost contenionat n decubit lateral,
efectund puncia pe faa anterioar i uor lateral articulaiei.
Dovada efecturii punciei a constat n exteriorizarea lichidului sinovial.
Pentru toate articulaiile s-a efectuat o aspiraie a lichidului, pentru a putea
fi nlocuit cu substana de contrast.
Injectarea substanei de contrast a fost efectuat lent n max. 20 de
secunde.
Expunerile cu raze X au fost efectuate cu ajutorul unui aparat SK-7-3
Medicor, din inciden latero-lateral i ventro-dorsal. Intervalele de expunere au
fost stabilite imediat dup administrare, dup 30 minute i la 24 h, animalele fiind
sub observaie clinic pentru evaluarea strii generale.

REZULTATE I DISCUII

Informaia obinut n urma examenului radiologic asupra articulaiilor este


important n modul de orientare a conduitei terapeutice, motiv pentru care
trebuie respectate toate etapele execuiei.
Realizarea punciei articulare a constituit n general cea mai dificil etap n
realizarea artrografiei, fiind posibil sub control radiologic sau prin evidenierea
lichidului sinovial.
Pentru stabilirea dozei utile, am fcut mai multe tatonri pentru a stabilii
cantitatea de substan ce poate fi administrat ntr-o articulaie, punndu-se n
eviden opacifierea spaiului articular i gradul de distensie a fundului de sac.
Substana s-a administrat cu grij, pentru a evita ruperea capsulei articulare ce ar
fi dus la diseminarea substanei periarticular, compromind investigaia.
Gradul de opacifiere a spaiului articular, precum i a elementelor articulare
au depins de tipul de substan folosit i de diluia acesteia.
n general, Odistonul introdus n spaiul articular a produs opacifierea pe o
durat variabil n timp, claritatea imaginii fiind mai bun imediat dup
administrarea soluiei, diminundu-se progresiv la 15 minute de la administrare,
n timp ce contrastul i calitatea imaginii obinute n urma administrrii de
Ultravist 300 au fost excelente n toate cazurile, cu opacizarea corespunztoare a
spaiilor comune, precum i contrast detaliat.
A existat doar un singur caz n care rezultatul a fost considerat slab, din
cauza diluiei rapide a mediului de contrast. Calitatea imaginii cu Ultravist 300 a

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fost foarte bun la 30 minute, comparativ cu a Odistonului, care a fost mult mai
diminuat.
Tolerabilitatea a fost diferit, la injeciile intraarticulare. Astfel injeciile
intraarticulare cu Odiston au fost dureroase, iar o parte din pacieni au prezentat o
inflamaie local.

Fig. 1 - Poziia corect a acului n cavitatea


Fig. 2 - Imagine radiologic a articulaiei
articular. Ultravist 300 expunere latero-
cotului expunere medio lateral n poziie
lateral la 5 minute articulaia scapulo-
neutr (Ultravist 300) la 10 minute
humeral

Fig. 3 - Artrografia umrului expunere din Fig. 4 - Imaginea radiologic a articulaiei,


inciden medio lateral, obinut prin expunere din inciden medio lateral
injectare de Odiston. Expunere la 10 minute (Odiston). Expunere la 20 minute

n cazul administrrii de Ultravist 300, injectarea intraarticular nu a fost


dureroas i nu s-au pus n eviden reacii sistemice imediate. Reaciile locale au

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fost absente. Imagini bune s-au obinut att n cazul folosirii de substane ca atare
ct i de substane diluate.
Dozele folosite au fost obinute prin tatonare, iar pentru a pune n eviden
diferitele componente ale articulaiei am recurs la expuneri multiple n ceea ce
privete incidena, ct i a razelor osoase ce contribuie la structura articulaiei.
Pentru un singur caz am utilizat diluia de 1:3, situaie n care s-au pus n
eviden imagini radiologice de calitate slab.

CONCLUZII

Utilizarea Ultravistului 300 n artrografii este mult mai bine tolerat,


comparativ cu preparatele clasice iodate (Odiston), chiar n cazul unor
concentraii i doze mai mari, iar frecvena i intensitatea reaciilor secundare au
fost mult diminuate.
Asocierea substanei de contrast cu anestezice locale a redus intensitatea
reaciilor secundare dureroase n momentul injectrii substanelor.
Concentraia optim de substan activ de contrast non ionic este ceea
de 0,623 g iopromid/ml din produsul comercial Ultravist 300.
Dozele utilizate n artrografii difer n funcie de talia animalului i de tipul
de articulaie dup cum urmeaz:
Pentru articulaia scapulo humeral s a utilizat 2,0 2,5 ml la cini de
talie mare i 1,5 ml la cini de talie mijlocie.
Pentru articulaia femuro tibio rotulian: 2,5-3 ml la cinii de talie mare
i 2,0 ml la cini de talie mijlocie.
Pentru a articulaia humero radio ulnar: 2,0 ml la cine talie mare i
1,5 ml la cini de talie mijlocie.
Artrografia este mai valoroas la articulaiile prevzute cu capsul
articular, cu condiia utilizrii de concentraii i doze adecvate, n vederea
opacifierii moderate a spaiului articular, oferind posibilitatea diferenierii pe
imagine a structurilor articulare.
Calitatea opacifierii n artrografie se reduce pe msur ce crete intervalul
de la injectare, perioada optim fiind de pn la 5 minute, precum i de tipul de
preparat utilizat. Preparatul non ionic Ultravist 300 produce o opacifiere de
lung durat.
Examinare fizic a lichidului sinovial confirm tolerana mai bun a
organismului la preparatul non ionic comparativ cu cel ionic.
Preparatul Ultravist 300 prezint reale avantaje n investigaiile
artrografice, ns are un foarte mare dezavantaj, acela al preului ridicat. Atunci
cnd nu se respect cu strictee modul de administrare i doza, substana poate
extravaza din articulaie ca urmare a ruperii capsulei articulare i s mascheze prin
opacifiere unele elemente articulare: de tipul fundurilor de sac, ligamentele
articulare sau unele formaiuni tendinoase.

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BIBLIOGRAFIE

1. Bonati F., A. Lombardo, L. Moschini, 1987 Non ionic contrast media Preclinical
and clinical data on Iopamidol Updated to december 1987, Milano, Centro studi
Bracco.
2. Damian A., I. Papuc, I. Chirilian, 2001 Anatomia comparat. Sistemul de
susinere i micare, Cluj-Napoca, Ed. AcademicPres.
3. Railhac I.I., I. Putois, 1983 Experimentation de liopamidol en arthrographie
opaque. Ann Rdiol 26: 812-816, 1983.
4. Ulici-Petru I., 1996 Contribuii privind utilizarea substanelor de contrast radiologic
la cine. Teza de doctorat U.S.A.M.V.
5. http://www.farmaline.ro/index.php?cid=5&pid=4394&&04ceddaf7cc18f3a8575a2417b
87e774=78374487

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OBSERVES REGARDING FELINE UROLOGIC


SINDROME INCIDENCE
OBSERVAII PRIVIND INCIDENA SINDROMULUI UROLOGIC FELIN

Luminia Diana HRICU


U.S.A.M.V. Iai

n ara noastr s-a observat n ultimii 10 ani o cretere a cazurilor


diagnosticate cu afeciuni urinare n special la felinele domestice.
Prin studiile efectuate ne-am propus s observm creterea
procentual a cazuisticii cu sindrom ulrologic felin i s stabilim ce loc
procentual ocup fa de celelalte afeciuni constatate la felinele domestice.

Key words: Urological sindrome, domestic feline, incidence

In the last 15 years in special literature it was observed frequently


urolithiasis in pets.
In our country in the last 10 years it was observed an increased of
frequency of urinary affections at domestical feline, especially an important
increased by year to year.
We proposed to observe what percent have urinary affection comparing
with others affections diagnosed.
Regarding to analyze urinary affections incidence, it was made
observations for more pathological entities diagnosed at the urinary apparatus.
We must mentioned that all three pathological entities in study are in
urologic feline syndrome (cystitis, uretritis, obstructive uropathy caused by
crystals, sand, real calculi, and urethral stenosis.

MATERIAL AND METHODS

The studies were made at Medical Clinic from Faculty of Veterinary


Medicine Iasi and at an Private practice in a period of 8 years meaning 1996
2003.
It was studied all domestic feline diagnosed with urinary affections and was
named at start Feline Urologic Syndrome and then it was made comparative
percentages between them and other affections diagnosed in the same period.
It was studied all cases male and female came for consultation in the
period 1996 2003 and diagnosed with hemorrhagic cystitis, urolithiasis or
spastic retention.
In the study period it was consulted 2057 domestically feline.

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RESULTS AND DISCUSSIONS

The casuistic offer by Medical Clinic and Private practice shoed that
incidence of urinary affections at domestic feline it was by 310 cases, registered into
1996 2003.
Regarding the incidence of affections by years repartition, dates are
evidenced in table 1.
Table 1
Incidence of urinary disturbances F.U.S. in domestic felines
in the period 1996 2003

Years 1996 1997 1998 1999 2000 2001 2002 2003 Total
No. cases 15 45 18 18 32 44 41 97
% comparing 310
4.83 14.51 5.80 5.80 10.32 14.19 1.22 3.28
by total

Analyzing the dates from table1, it was observed that the most rich year in
casuistic was 2003, when was diagnosed 97 cases with feline urological syndrome
who represent 31,28% by total.
High frequency it was registered in 1997 too which represent 14,51%.
In the next years the incidence was low, meaning in 2001, 2002, and 2000,
when was registered approximately the same number of cases. So then, in 2001
are mentioned 44 cases who represent 14,19 percent by total, in 2002, was
registered 41 cases domestic feline diagnosed, who represent 13,22% by total and
in 2000 was diagnosed 32 cases number who represent 10,32% by total (310
cases).
In the 1998 and 1999 years it was observed a coincidence of number of
cases, so the prevalence in itch of them is 18 cases, meaning 5,80% by total.
1996 is the year with the lower number of cases diagnosed, 15 cases who
represent 4,83% by total.
Referring to urinary affections incidence in the period for study (1996
2003), excepting 1997, where the prevalence is high, it was observed a real
increased of number of cases. That might by due the one side, by increase of the
owners of animals and of course the increase of domestic feline population for
pets and by the other side the increase of interest of owners for the health of pets.
Another reason might be the explosion of the different sorts of food (especially
dry) and adopted of them by owners (the easiest food for animals).
Knowing the incidence of urinary affections by each year we tried to
determined the percentage of urinary affections comparing of domestic feline
population with another medical affections in every year and finally in 8 years of
study.

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Table 2
Incidence of F.U.S. (feline urologic syndrome) cases vs. domestic feline population
under treatment in the period 1996-2003

% FUS comparing
Total number of No. cases
Year with total no. of
cases presented with FUS
cases
1996 248 15 6.04
1997 325 45 13.84
1998 126 18 14.28
1999 98 18 18.36
2000 149 32 21.47
2001 240 44 18.33
2002 298 41 13.75
2003 573 97 16.92
Total 2057 310 15.07

Analyzing table 2 it was observed that highest percent of urinary affections


it was in 2000 year, when it was consulted and diagnosed with different medical
affections 149 domestic feline. By them, 32 was diagnosed with one by three
pathological entities from study who represent 21,47 percent.
In decrease is 1999 year when by total of 98 domestic feline consulted, 18
was diagnosed with feline urologic syndrome, who represent 18,36 percent.
In 2001 and 1999 years, the percents are equal, but in 2001 the number
total of cases presented for consult was highest, meaning 240 cases, and urinary
affections was diagnosed in 44 cases from total.
2003 year was the most rich in casuistic (573 feline) and by them 97 was
diagnosed with feline urologic syndrome, number represented in 16,96 percent.
In 1998 it was consulted 126 domestic feline and by them 18 was
diagnosed with urinary affections (meaning 14,28%).
1997 was an urologic casuistic reach year, 45 cases, but low percentage is
due to high number of consulted cases of domestic feline.
The next was 2002 year, with a decreasing percentage (13,75) similar with
1997, so it was consulted 298 pets and by them 41 was studied.
The year with the low cases diagnosed with urinary affections was 1996,
when was consulted 248 domestic feline and by them only 15 meaning 6,04% had
urinary affections.
Making a rapport of consulting in this period (2057) we can say that FUS
was diagnosed in 15,07%.

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CONCLUSIONS

In the period of study it was observed an increase of cases diagnosed with


Feline urological syndrome starting at 4,83% in 1996 to 31,28% cases diagnosed
in 2003.
Comparing with other types of affections in 8 years of study it was
observed a increase of FUS starting 6,04% in 1996 and an peaks of 21,47% in
2000.
After our studies in 8 years it was observed that incidence of cases with
feline urologic syndrome have a 15,07 percent from consulting feline.

BIBLIOGRAPHY

1. Brz H., 1981 Patologie i clinic medical veterinar. Ed. Didactic i


Pedagogic, Bucureti.
2. Cotard, J.P., 1992 L'urolithiase feline. Le Point Vet., vol. 24, nr. 146, aug. sept
3. Ettinger S.J., 1983 Disease of the dog and cat, Textbook of veterinary internal
medicine vol II, Ed bz W.B. Saunders Company Philapelphia USA.
4. Ghergariu, S., 1995 Patologia nutriional i metabolic a animalelor. Ed.
Medical Vet. Bucureti.
5. Hagiu, N., 1993 Patologie medical veterinar. Curs Lito. Vol. 2. C.M. Institutul
Agronomic Iasi, FMV
6. Mihai D., 1994 Patologie i clinic medical veterinar. Ed. Ceres, Bucureti., Vol.
2; Bolile aparatului urinar.
7. Osborne C.A., Finco D.R., 1977 - Urinary tract infections: New solutions to old
problems. In Scientific Proceedings of the 44 th Annual Meeting of the American
Animal Hospital Association. South Bend, Indiana, Am Anim. Hosp. Assoc.

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DIAGNOSTIC ASPECTS REGARDING ESTABLISH TYPE


OF UROLITHIASIS TO A DOMESTIC FELINE
(CASE STUDY)
ASPECTE DE DIAGNOSTIC N VEDEREA STABILIRII TIPULUI DE UROLITIAZ
LA O FELIN DOMESTIC (STUDIU DE CAZ)

Luminia Diana HRICU, Gabriela RILEANU


U.S.A.M.V. Iai

n lucrarea de fa ne-am propus un studiu de caz pentru a


evidenia importana corectitudinii diagnosticrii urolitiazei.
Pentru stabilirea exact a diagnosticului de urolitiaz s-au utilizat
pe lng examinarea clinic, o serie de examene suplimentare de
diagnostic (ecografia, radiografia, microscopia optic i microscopia
electronic).
n urolitiaz este foarte important de stabilit att tipul de cristalurie
sau de urolit ct i nivelul localizrii calculozei (renal, ureteral, vezical sau
uretral). Doar dup stabilirea unui diagnostic cert se poate recurge la
efectuarea unui tratament n funcie de tipul de urolit.

Urolithiasis is known affection at animals and in human. In speciality


literature the urolithiasis is considered disease of century.
After last study made in our country regarding the incidence it may say that
urolithiais at the domestic feline is very frequent so, at consultation are presented
numerous cases male and female.

MATERIAL AND METHODS

The study was effected in a cat, commune breed, in 7 year age, with
thenesma, disury, frequent urinate and low quantity, urine colour was red.
In attentive examination it was observed that the cat was eliminated calculi
in urinate time on the consult table.
The exam was made by know methods (observation, inspection, palpation).
For confirm the urolithiais it was used secondary means of diagnosis,
especial ultrasonography, and radiography.
For a good therapeutically decision it was harvest the urine (for the
sediment exam).
It was executed a simple test with strip type DEKA-PHAN-LEUCO.
Because in sediment were small calculi (eliminated by natural way) it was
exam almost the sediment and the small calculi at optic microscope and then just
calculi at scanning microscope electronic.
The sediment it was obtained from centrifuge of fresh urine.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

It was exam at microscope between two lames, without coloration, or with


simple coloration with metal blue or iron tincture.
A few probes were exam at scanning microscope electronic for evidence
the structure of stone surface. It was used the scanning microscope electronic
TESLA BS 300.
The clarification was made with aetilic alcohol solution 980, then the probes
it was fixed in aluminum cylindrical supports (using a paint with conduction
proprieties), and after that it was introduced in methalisor.
The methalisor method consist in silver evaporation in vide and set down
the atomic powder of an angle on the probe. The metal set down layer had a
great power to spread the electrons which ensure the contrast wish of image.
The tension of acceleration of electrons was till 20 kV

RESULTS AND DISCUSSIONS

At the consult it was observed the attitude for urinate frequent taken by cat,
at palpation of bladder it was observed the pain express by long mewed and a low
urine quantity of red colour in which was very small calculi.
At the exam of urine with strip it was observed a pH-8, abundant urinary
sediment, urinary density 1.030, high number of hematites, and high proteins.
Ultrasonography evidenced a large bladder wall, abundant urinary
sediment, a calculus with 12 mm dimension and numbers small calculi.
At radiography was evidenced a radio opaque addition in the bladder.
By sediment exam at optical microscope it was evidenced the existence of
hematies, leukocytes, the I and II level cells and the crystals of struvite and
calcium carbonate.
To the scanning microscope electronic image of fragments of calculi it was
observed that the sediment contained highest struvites and fracture of calculus
contained a lot of calcium carbonate crystals.

Fig. 1 - Scanned image of fraction surface Fig. 2 - Scanned image of fraction surface
of calcium carbonate calculi x 69 of calcium carbonate calculi x 170

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Fig. 3 - Scanned image of fraction surface Fig. 4 - Scanned image of fraction surface
of calcium carbonate calculi x660 of calcium carbonate calculi x 1260

Fig. 5 - Scanned image of fraction surface Fig. 6 - Scanned image of fraction surface
of calcium carbonate calculi x 2400 of calcium carbonate calculi x 8000

CONCLUSIONS

After clinical examination and testing the urine with strip it can establish
only the diagnostic of Feline urologic syndrome (FUS). Only supplementary tests
for diagnostic (ultrasonography and radiography), can establish existence of
urolithiasis.
Identification of crystals type or a type of calculi is necessary for establish
efficacious therapy.
Unorganised elements who composed the urine are represented by hasten
salts crystalline or amorphous form. Always, crystallizations depend by urine
concentration and by the urinary pH. Alkaline reaction favour crystallisations of
struvites and calcium carbonate.
To identification the calculi had a bimineral mixture meaning ammonium
magnesium phosphates and calcium carbonate, both of them crystallising to
alkaline pH.

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BIBLIOGRAPHY

1. Codreanu M., Diaconescu A., 2003 Diagnosticul ecografic al animalelor


de companie. Ed. Coral Sanivet Bucureti.
2. Cotard J.P., 1992 Lurolitiase feline. Le Point Vet .vol 24, nr.146, aug-sept.
3. Ghergariu S., 1988 Patologie medical a animalelor. Ed Ceres Bucureti
4. Hesse A., Sanders G., Leusmann D.B., 1986 Analyses of feline urinary
stones using infrared, spectroscopy and scaning electron microscopy.
Kleintiepraxis 35:10,505 512.
5. Hricu Luminia Diana, Vulpe V., 2001 Aspecte privind metodologia
diagnosticului afeciunilor aparatului urinar la cine i pisic. Lucr t.vol
44 seria MV, p 340.

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ASPECTS OF ENZYMATIC DIAGNOSIS


OF HEPATO-BILIARY DISEASES IN DOG
ASPECTE ALE DIAGNOSTICULUI ENZIMATIC
N AFECIUNILE HEPATO-BILIARE LA CINE

R.M. SAUCIUC, N. HAGIU, Gh. SOLCAN,


M. MUSTEA, V. BOGHIAN
U.S.A.M.V. Iai

The goal of this study was to establish specificity and sensibility of


serum enzymes with hepatic primary origin in diagnosis of hepato-biliary
diseases in dog and also the influence of induction of enzymatic activity
produced by treatement with phenobarbital. Serum biochemical tests
accomplished on dogs with hepato-biliary diseases reveald a 100%
sensibility of AST (aspartate aminotransferase) and also a relative
sensibility of ALT (alanine aminotransferase), ALP (alkaline phosphatase)
and GGT (gamma glutamyl transferase). The study revealed also a high
specificity of all four enzymes(ALT, AST, ALP, GGT) demonstrated by
significant increases of mean values for each of these.
In epileptic syndrome cases the influence of phenobarbital treatment
on enzymatic serum activity was minimum, mean values of ALT, ALP and
GGT were in reference range. A relatival high sensibility of AST serum
activity was observed at 6 from 10 dogs treated with phenobarbital but
these increasing were moderate indicating a possible induction of enzymes
activity or an extrahepatic origin of serum AST.

Key words: dog, hepato-biliary enzymes, enzymatic induction, phenobarbital

The main serum enzymes included in routine tests of hepato-biliary


diseases in dog are reperesented by: liver transaminases (ALT- alanine
aminotransferase, AST aspartate aminotransferase), ALP alkaline phosphatase
and GGT gamma glutamyl transferase.
After Bain P.J. (2003) the tissues with highest activity of ALT at animals
are liver and muscular tissues (cardiac and skeletic). Certain researches showed
that in dogs the highest ALT activity is in hepatic cells unlike the ruminants, pigs,
horses and birds hereupon serum ALT activity originated from the liver is very
low. In this way increasing in serum ALT activity are considered relative specific
for liver diseases. The mechanism of increasing of serum ALT activity include
both enzyme releasing folowing cells injuries and induction of enzyme activity
(increase of ALT synthesis) folowing treatments with certain medicines
(glucocorticoids, anticonvulsivants). Plasma half life of ALT in dogs is about 60
hours. (9)

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AST is present in high concentrations in a great variety of tissues. In dog


the tissues with highest concentrations of AST are: heart, liver, skeletal muscle
tissue, kidney and brain. Plasma half life of AST in dog is about 12 hours. (5)
ALP is associated with cells membranes in many tissues. Certaines
isoenzymes named by their origin are known in this way. Just three of these
isoenzymes have clinical meaning, these contributing widley at serum activity of
ALP: bone isoenzyme (BALP), liver isoenzyme (LALP), corticosteroids induced
isoenzyme (CALP). (8)
CALP is present in low concentrations in serum but high levels can be
induced by exposure at high concentrations of exogenous and endogenous
glucocorticoids. LALP and CALP originate from portion of hepatocytes
membrane and biliary channels. BALP have osteoblastic origine and her functions
are not completely elucidated.
GGT is known also by her old name of gamma glutamyl transpeptidase. In
dog the tissue with highest concentrations in GGT is kidney. In this case liver
have a low concentration of GGT. Because of preponderant urinary excretion of
kidney-originated GGT is supposed that major source of GGT serum activity is
represented by liver. Same as ALP, increasing of serum GGT is a result of de
novo liver synthesis than from release of free enzyme from cytoplasm.

MATERIALS AND METHODS

The study was made on clinical cases from Clinic of Internal Medicine from
Faculty of Veterinary Medicine of Iasi and included analyses of serum enzymes
activity on 30 dogs divided in three categories: ten healthy dogs, ten dogs with
different hepato-biliary diseases and ten dogs with epileptic symptoms treated at
least two month with phenobarbital.
Investigations consisted in biochemical analyses of serum regarding
serum enzymes activity and certains parameters which reflect the activity of
hepatobiliary system. In this way ALT (alanine aminotransferase), AST (aspartat
aminotransferase), ALP (alkaline phosphatase), GGT (gamma glutamyl
transferase), BT (total bilirubin), BUN (blood ureea nitrogen), PT (total proteins),
CHOL (total cholesterol), TGL (triglycerides) were analyzed by using of semi-
automatc biochemical analyzer. EOS 880 Plus.

RESULTS AND DISCUSSIONS

Since variation of serum enzymes activity limits are different from an


researcher to another and in this situation is difficult to establish exactly the
maximum and minimum limit having pathological significance, was necessary to
establish the normal values of our cases and to compare these with those from
literature. Dogs selected for this category were represented by healthy animals,
from different breeds, between 1,5 and 12 years old. We avoided to introduce in

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this group dogs with with age below 1 year hereupon serum values of ALP are
influenced by osteoblastic originated isoenzime (BALP).
The results obtained on healthy dogs are presented in tabel 1 and revealed
that values of serum enzymes activity are quite similarly with those from medical
literature. (1, 4)
Tabel 1

ALT AST ALP GGT BT BUN PT CHOL


Nr.
U/L U/L U/L U/L mg/dl mg/dl mg/dl mg/dl
1 73,1 23,0 41,2 2,3 0,4 23,0 7,0 145,0
2 25,6 16,2 28,0 4,1 0,2 12,7 6,8 231,2
3 40,0 50,2 21,3 3,2 0,2 14,8 6,4 248,0
4 22,0 41,6 124,0 5,4 0,3 25,1 5,9 186,8
5 18,0 58,5 90,6 1,4 0,1 37,0 7,7 151,1
6 86,0 42,0 44,3 1,6 0,1 22,0 7,2 221,4
7 71,8 53,1 81,0 3,2 0,3 48,9 6,0 174,4
8 26,7 69,3 34,1 6,0 0,6 52,0 7,1 220,7
9 92,0 18,4 61,8 1,6 0,3 16,4 7,6 192,3
10 54,3 19,2 74,0 2,8 0,1 42,1 7,2 262,0
Mean values 50,95 39,15 60,03 3,16 0,26 29,4 6,89 203,29
obtained 28,13 18,94 34,42 1,6 0.157 14,47 0.61 39.72
Reference 1,2- 5,5- 125-
5-102 10-55 20-156 0,1-0,3 15 - 50
ranges* 6,4 7,5 250
*(Aielo ES et all, 1998, Ghergariu S et all, 2000

The second category of dogs studied was represented by ten dogs from
different breeds and with different age and sexes which were diagnosed with
some primary or secondary hepatobiliary diseases (table 2).
Table 2

Nr.
Breed and age Diagnosis
crt.
1 Dog German sheepard 10 years Acute hepatitis
2 Dog Pekinez 4 years Jaundice -Gallblader stone
3 Bitch Crossbreed 4 years Liver cirrhosis, Liver steatosis
4 Dog Crossbreed 10 years Liver carcinoma - jaundice
5 Dog German Sheepard 10 years Limfosarcoma difuse liver localization
6 Dog German sheepard 9 years Cervical adenocarcinoma liver congestion
7 Bitch German sheepard 12 years Liver cirrhosis ascitic syndrome
8 Dog Crossbreed 11 years Chronic active hepatitis
9 Bitch Akita Inu 5 years Chronic hepatitis
10 Dog German sheepard 10 years Limfosarcoma nodular liver localization

Results of these serum analyze are presented in table 3. A high sensibility


of AST wich was observed it increasing in 100% from cases of hepato-biliary

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diseases. Also AST demonstrated a high specificity for hepato-biliary diseases,


its mean value increasing five time normal.
A relative high sensibility was noticed in case of ALT wich increased in 8
from 10 cases (excepting cases of liver cirrhosis and gallbladder stone). Mean
value observed for those 10 cases increased four time normal showing a high
specificity for hepatobiliary diseases.
Values of ALP demonstrated significant increases in 8 from 10 cases
(excepting liver congestion and chronic hepatitis cases) showing a relative high
specificity. Though mean value for those 10 cases for ALP increased two time
normal demonstrating a high specificity for liver diseases.
GGT showed a relative high sensibility, it serum activity increasing in 7
from 10 cases (excepting acute hepatitis, liver congestion, and chronic hepatitis).
The mean value increased five time normal demonstrating a high specificity of
GGT for liver diseases in dog.
Table 3

ALT AST ALP GGT BT BUN PT CHOL


Nr.
U/L U/L U/L U/L mg/dl mg/dl mg/dl mg/dl
138,1 306,0 162,4 0,793 43,82 95,0 6,4 151,7
57,41 225,7 218,8 7,394 41,79 32,0 6,1 221,5
87,75 523,6 145,6 13,32 5,037 41,06 5,0 168,9
866,8 613,2 272,9 134,0 7,9 11,74 8,0 322,9
206,7 214,5 327,2 6,615 1,022 47,46 6,5 158,5
343,4 256,0 6,145 5,12 0,589 61,67 7,8 256,6
442,2 146,4 365,9 18,54 0,392 14,38 5,4 127,3
900,7 181,9 154,8 90,48 3,453 56,06 11,0 413,1
234 143,2 169,8 5,2 0,281 67,45 6,2 278,0
952,9 371,0 495,0 51,61 2,508 72,0 4,5 176,0
Mean values 422,99 298,15 231,85 32,80 10,67 49,88 6,69 227,45
353,14 167,38 143,42 45,19 17.10 26.42 1.87 90,43
Reference 5,5- 125-
5-102 10-55 20-156 1,2-6,4 0,1-0,3 15 - 50
ranges * 7,5 250
*(Aielo ES et all, 1998, Ghergariu S et all, 2000)

Results of analysis on the third category of dogs showed a minimum


influence of treatment with phenobarbital on serum enzymes activity. In this way
six cases of slowly increases of AST value were noticed and one case with
increased GGT value. Mean values in 3 from 4 cases were between normal ranges
(tabel 4). Only mean value for AST was moderate increased indicating a possible
process of enzymatic induction or a extrahepatic origin of serum enzyme.

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Tabel 4

ALT AST ALP GGT


Nr. Sex, breed, age
U/L U/L U/L U/L
1 Dog-Bichon Malthese-2,6 years 56,7 75 43,16 1,8
2 Bitch -German shepard-3 years 86,5 206,5 23,0 2,06
3 Bitch - Pekinez-10 years 43,11 68,98 67,12 1,74
4 Dog - German shepard -6 years 22,41 53,87 26,40 3,2
5 Dog - Crossbreed 4 years 16,02 38,0 23,09 4,5
6 Dog -Boxer-2,5 years 50,04 97,1 43 8,0
7 Bitch - Crossbreed -4,6 years 77 21 102 3,0
8 Dog Pekinez 8 years 80,5 163 128 5,0
9 Dog -Doberman-6 years 16 41 94 2,22
10 Dog Husky- 2 years 69 74 36.07 3,2
51,72 83,84 53,63 3,47
Mean values obtained
33,15 58,37 39,52 1,93
Reference ranges* 5-102 10-55 20-156 1,2-6,4
*(Aielo ES et all, 1998, Ghergariu S et all, 2000)

CONCLUSIONS

1. In hepato-biliary diseases liver enzymes presents a high specificity


demonstrated by significant increasing of ALT, AST, ALP and GGT mean
values.
2. Sensibility for those four liver enzymes was relatively high, at least two
from four enzymes beeing increased in each case. A special sensibility was
demonstrated by AST which increased values in all these ten cases of hepato-
biliary diseases. Surprisingly in some severe injury like liver cirrhosis and
gallbladder stones ALT values were between reference ranges. We conclude that
one set of analysis is helpful for diagnosis only when are used all those four
enzymes together, otherwise there is a great risk of error in diagnosis.
3. The influence of phenobarbital treatment on serum enzymes activity is
reduced, only AST demonstrated a moderate sensibility but in this case increasing
can be explained by possible enzymatic induction or by an extrahepatic origin of
serum AST.

BIBLIOGRAPHY

1. Aielo E. Susan et all., 1998 - The Merck Veterinary Manual, Eighth edition, National
Publishing, Inc. Philadelphia, Pennsylvania.p.2192-2193
2. Bain P.J. in: Latimer K.S., Mahaffey E.A., Prasse K.W., 2003 - Duncan and
th
Prasses Veterinary Laboratory Medicine: Clinical Pathology, 4 ed. Ames, Iowa
State Press, p. 193 214.

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3. Center S.A., Slater M.R., Manwarren T., Prymak K., 1992 - Diagnostic efficacy of
serum alcaline phosphatase and gamma glutamyltransferase in dogs with
histologically confirmed hepatobiliary disease: 270 cases (1980-1990). J Am Med
Vet Assoc vol. 201 p. 1258-1264
4. Ghergariu S., Pop A., Kadar L., Spnu Mariana, 2000 - Manual de laborator clinic
veterinar, Editura ALL Educaional Bucuresti.
5. Meyer D.J., Harvey J.W., 1998 - Veterinary laboratory medicine: Interpretation and
diagnosis. Philadelphia W.B.Saunders Co. p. 157 186
6. Muller P.B., Taboada J., Hosgood G., Partington P.B., VanStenhouse J.L.,
Taylor H.W., Wolfsheimer K.J., 2000 - Effects of long-term phenobarbital
treatment on the liver in dogs. J Vet Intern Med vol. 14, p. 165 171
7. Solcan Gh., Boghian V., Rollin F., 2005 - Patologie i clinic medical veterinar,
Edit. Ion Ionescu de la Brad Iai
8. Sykalima M., Takiguchi M., Yasuda J., Hashimoto A., 1997 - The age dependent
levels of serum ALP isoenzymes and the diagnostic significance of
corticosteroid induced ALP during long-term glucocorticoid treatment. J Vet Med
Sci vol. 59(10), p. 905 909
9. Stockham S.L., Scott M.A., 2002 - Fundamentals of Veterinary Clinical Pathology.
Ames, Iowa State University Press, p. 434 - 459

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Universitatea de tiine Agricole i Medicin Veterinar Iai

STUDIUL EFECTELOR APITERAPIEI CU PRODUSUL


HEPRO N AFECIUNI HEPATICE LA CINE
A STUDY ON HEPRO PRODUCT API-THERAPEUTICAL EFFECTS
IN DOG LIVER DISEASES

I. RDOI1, Agripina APCALIU2


Carmen NEGOI1, Crengua PAVEL1
D. CONDUR3
1
U.S.A.M.V. Bucureti
2
I.C.D.A. Bucureti
3
Universitatea Spiru Haret Bucureti

The worldwide increasing interest on alternative and complementary


medicine, especially on veterinary api- and phyto-therapy, encouraged us to
research the HEPRO api-therapeutic product effects in some dog liver
diseases.
Unless classical phyto- and api-therapy, which uses the brute bee
products, our modern api-phyto-therapy uses the product HEPRO, made
and conditioned by ICDA in co-operation with Veterinary Faculty from
Bucharest, following the researches made in 2002-2006 period.
The healthy effects on internal organs and tissues were observed
after six weeks treatement with HEPRO product, in miscellaneous liver
diseases (minor liver failure, cholestatic diseases, acute hepatitis, liver
cirrosis).
HEPRO nutritional therapy methods applied in presented liver
diseases had at least the same effectiveness as standard therapeutical
methods.

MATERIAL I METOD

Investigaiile clinice i examenele ecografice i de laborator au fost


realizate n Clinica de Medical a Facultii de Medicin Veterinar Bucureti,
selectndu-se 17 cazuri pentru experiment.
Cercetrile s-au realizat pe cini cu vrste i rase diferite (tabel 1, 2), cu
diverse afeciuni hepatobiliare, care au fost tratai prin mijloace terapeutice
clasice, comparativ cu protocolul terapiei nutritionale cu produsul HEPRO.
Examenele de laborator au urmrit n principal att determinarea
bilirubinemiei, a activitii transaminazelor (ALAT, GT) i fosfatazei alcaline
(PAL) din snge pe Reflotron (chituri uscate) ct i efectuarea examenului
complet al urinei (COMBUR-TEST). Testele au fost repetate sptmnal.
Valorile normale de referin la cine pentru parametrii determinai n
laborator au fost urmtoarele: bilirubinemie: < 0.61 mg/dl; transaminaza ALAT: <
40 U/l; transaminaza GT: <5 U/l; fosfataza alcalin (PAL): 20-150 U/l;
proteinemia 5,4-7,7 g/dl; albuminemia 2,5-3,7 g/dl; bilirubinuria +.

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Tabel 1
Afeciunile hepatice diagnosticate la cini i protocolul terapeutic aplicat

Afeciune Tratament medicamentos Apiterapia cu HEPRO


Hepatit acuta Cazul 1 Cazurile 11, 13
Colestaz Cazul 4 Cazurile 8, 10, 12
Mica insuficien hepatic Cazul 3 Cazurile 6, 14, 17
Ciroza hepatic Cazul 2 Cazurile 15, 16

Tabel 2
Apiterapia cu produsul hepro,dozele zilnice

Mica insuf.
Specificare Colestaz Ciroz hepatic
hepatic
Cini talie mic 3cp/10kg 4cp/10kg 4cp/10kg
Cini talie mijlocie 2cp/10kg 2cp/10kg 3cp/10kg
Cini talie mare 1cp/10kg 2cp/10kg 3cp/10kg

Protocolul terapeutic a fost instituit n funcie de fiecare caz, iar pentru


pentru apifitoterapie a fost utilizat produsul HEPRO, rezultat al cercetarilor pe o
perioada de peste 3 ani, n doze de 1-4 cp/10 kg greutate vie, zilnic, n trei pro
dosis, cu 30-60 minute nainte de mas, timp 44 zile.

REZULTATE I DISCUII

Rezultatele cele mai spectaculoase ale apiterapiei cu produsul HEPRO


s-au constatat la cini cu mic insuficien hepatic, unde analizele au artat
revenirea la normal a valorilor bilirubinemiei i transaminazelor, cu rezultate
superioare chiar, tratamentului medicamentos.
n colestaz, rezultatele tratamentului cu produsul HEPRO sunt de
asemenea superioare tratamentului medicamentos, ducnd la o normalizare a
parametrilor biochimici studiai.
n hepatita acut i n ciroza hepatic, rezultatele analizelor de laborator
dup 44 zile de tratament, arat o tendin accentuat de vindecare, ns necesit
mai multe edine de apifitoterapie nutritionala cu produsul HEPRO, pn la
revenirea la normal a parametrilor biochimici investigai, n corelaie cu o evoluie
clinic favorabil ce caracterizeaz vindecarea.
Parametrii biochimici analizai arat o eficien, cel puin egal a
apiterapiei cu HEPRO, fa de terapia clasic n toate cele 4 tipuri de afeciuni
hepatice, rezultatele fiind prezentate n tabelele 3-6.
Rezultatele analizelor nu pot fi raportate la rezultatele altor experimente
efectuate pe cini afeciuni hepatice prin utilizarea apiterapiei, ns pot fi
comparate rezultatele obinute n urma apiterapiei cu produsul HEPRO, faa de
tratamentului medicamentos dup 44 zile de tratament.

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Tabelul 3
Rezultatele investigaiilor biochimice nainte de tratament i dup 44 zile de
tratament la cini cu mic insuficien hepatic

Tratament Apiterapia cu
Valoare iniial
medicamentos HEPRO
Parametru
Caz Caz Caz Caz Caz Caz Caz Caz Caz Caz
3 6 9 14 17 3 9 6 14 17
Bilirubinemie
0,7 0,9 1 1 0,8 0,6 0,5 0,3 0,3 0,5
(mg/dl)
ALAT (U/l) 78 80 85 83 69 67,8 60 51,5 41,2 33
PAL (U/l) 105 108 100 115 107 76 62 60 91 84

Tabelul 4
Rezultatele investigaiilor biochimice nainte de tratament i dup 44 zile de
tratament la cini cu colestaz

Tratament Apiterapia cu
Valoare iniial
medicamentos HEPRO
Parametru
Caz Caz Caz Caz Caz Caz Caz Caz Caz Caz
4 7 8 10 12 4 7 8 10 12
Bilirubinemie
1,6 2,1 1,1 0,8 2,5 0,8 0,85 0,6 0,5 0,85
(mg/dl)
ALAT ( U/l) 103,5 108 102 105 107 86,8 68 79 55 36
PAL (U/l) 107 112 105 60 115 64 58 41 53 60
GT (U/l) 11,24 12 10 10,2 13,3 9,8 7,5 4,1 6,4 4,2
Bilirubinurie +++ ++ ++ ++ +++ + + + + +

Tabelul 5
Rezultatele investigaiilor biochimice nainte de tratament i dup 44 zile de
tratament la cini cu hepatit acut

Tratament Apiterapia cu
Valoare iniial
medicamentos HEPRO
Parametru
Cazul Cazul Cazul Cazul Cazul Cazul Cazul Cazul
1 5 11 13 1 5 11 13
Bilirubinemie
2,7 2,2 3 2,5 2,2 1,9 0,85 1,1
(mg/dl)
ALAT ( U/l) 160 180 165 140 88 109 68 36
PAL (U/l) 105 130 107 105 77 88 69 85
GT (U/l) 11 12 12 11,3 9,1 8,1 9,4 8,7
Albuminemie
1,2 1,5 2,1 2,5 2,6 2,7 2,44 2,95
(g/dl)
Bilirubinurie ++ ++ ++ ++ + + + +

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Tabelul 6.
Rezultatele investigaiilor biochimice nainte de tratament i dup 44 zile de
tratament la cini cu ciroz hepatic

Tratament Apiterapia cu
Valoare iniial
medicamentos HEPRO
Parametru
Cazul Cazul Cazul Cazul Cazul Cazul
2 15 16 2 15 16
Bilirubinemie (mg/dl) 1 1,3 0,9 0,9 0,8 0,9
ALAT ( U/l) 102 105 98 56 59 56
PAL (U/l) 110 115 107 96 101 94
Albuminemie (g/dl) 1 1,3 2,5 2,95 2,4 2,8
Proteinemie (g/dl) 4 3,8 4,5 4,85 4,96 4,7
Bilirubinurie ++ ++ ++ + + +

Rezultatele investigaiilor biochimice (tabel 3) la cini cu mic insuficien


hepatic au artat dup 44 zile de tratament o revenire la normal a bilirubinemiei
(0,3-0,5 mg/dl), n urma apiterapiei cu produsul HEPRO, fa de tratamentul
medicamentos (0,5-0,6 mg/dl) i o scdere a activitii transaminazice ALAT (33-
51,5 U/l) fa de tratamentul medicamentos 60-67,8 U/l).
La cini cu colestaz investigaiile biochimice au artat valori apropiate ale
bilirubinemiei n urma tratamentului cu produsul HEPRO,de 0,5-0,85 mg/dl fa
de 0,8-0,85 mg/dl n urma tratamentului medicamentos.
Valorile activitii transaminazei GT au fost ntre 4,1-6,4 U/l n urma
apiterapiei, comparativ cu limitele 7,5-9,8 U/l n urma tratamentului
medicamentos. Transaminaza ALAT a sczut n urma apiterapiei la valori
cuprinse ntre 36-79 U/l fa de 68-86 U/l n urma tratamentului medicamentos.
Fosfataza alcalin (PAL) i bilirubinuria au sczut semnificativ prin ambele
procedee terapeutice, nct se poate concluziona c efectul apiterapiei cu
produsul HEPRO a fost cel puin la fel ca i terapia medicamentoas dup 6
sptmni de tratament.
La cinii cu hepatit acut, scderea bilirubinemiei la valori de 0,85-1,1
mg/dl n urma apiterapiei fa de 1,9-2,2 mg/dl n urma tratamentului
medicamentos, ct i scderea transaminazei ALAT la valori cuprinse ntre 36-68
U/l fa de 88-109 U/l n urma tratamentului medicamentos, ceea ce justific
utilizarea apiterapiei cu produsul HEPRO. Valorile albuminemiei, ale fosfatazei
alcaline, GT i bilirubinuriei fiind sensibil apropiate n ambele procedee
terapeutice, aceasta arat tendina accentuat spre vindecare ns necesit o
perioad de terapie nutritionala cu produsul HEPRO mult mai lung, pn la o
vindecare complet.
La cini cu ciroz hepatic, rezultatele apiterapiei arat o scdere a
activitii transaminazei ALAT pn la valori de 56-59 U/l (fa de 40 U/l care
este limita superioar a valorilor normale) cu o cretere semnificativ a
proteinemiei i albuminemiei ns sub limitele normale, concomitent cu scderea

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bilirubinei la 0,9 mg/dl (ns peste limita superioar a valorilor de referin 0,6
mg/dl), rezultate foarte apropiate de cele obinute n urma tratamentului
medicamentos.
Eficiena tratamentului medicamentos i a apiterapiei cu produsul HEPRO
n ciroza hepatic, nu poate fi demonstrat dup o perioad de 44 zile, astfel nct
investigaiile trebuie efectuate dup mai multe luni de zile de tratament. Exist
ns o tendin de revenire la normal a parametrilor biochimici sanguini i urinari,
corelat cu evoluia clinic favorabil a cazurilor urmrite n experiment.
Investigaiile trebuie efectuate pe un numr mai mare de animale i o
perioad de timp prelungit n cazul animalelor cu ciroz hepatic, ns rezultatele
analizelor biochimice sugereaz o posibil asociere a apiterapiei si fitoterapiei cu
tratamentul medicamentos (inclusiv perfuzii cu albumin, aminoacizi soluii
rehidratante).

CONCLUZII

1. Metodele de apifitoterapie cu produsul HEPRO n afeciunile hepatice


prezentate, au fost cel puin la fel de eficiente ca i metodele terapeutice standard.
2. La cini cu mic insuficien hepatic, dup 44 se constat revenirea la
normal a bilirubinemiei, sub 0,61 mg/dl n urma ambelor procedee terapeutice i
scderea activitii ALAT (33-51,5 U/l) n urma apiterapiei , fa de (60-67,8 U/l)
dup tratamentul medicamentos.
3. La cini cu colestaz, n urma terapiei cu produsul HEPRO,
bilirubinemia a sczut la 0,5-0,85 mg/dl, fa de 0,8-0,85 mg/dl n urma
tratamentului medicamentos, corelat cu scderea GT (4,1-6,4 U/l) fa de 7,5-
9,8 U/l prin tratamentul medicamentos i cu scderea ALAT (36-79 U/l) n urma
apiterapiei comparativ cu 68-86 U/l prin tratamentul medicamentos i cu scderea
fosfatazei alcaline i a bilirubinuriei prin ambele metode de tratament.
4. La cini cu hepatit acut, scderea bilirubinemiei la valori de 0,85-1,1
mg/dl n urma apiterapiei fa de 1,9-2,2 mg/dl prin tratamentul medicamentos
corelat cu scderea activitii ALAT (36-68 U/l) dup terapia nutritionala cu
produsul HEPRO fa de 88-109 U/l n urma tratamentului medicamentos,
justific utilizarea apiterapiei, artnd tendina de vindecare, demonstrat i de
corectarea albuminemiei, bilirubinuriei i a activitii enzimelor PAL i GT.
5. n ciroza hepatic la cine n urma apiterapiei produsul HEPRO se
constat scderea ALAT (56-59 U/l) i a bilirubinemiei (0,8-0,9 mg/dl) corelat
cu creterea foarte semnificativ a albuminemiei (2,4-2,8 g/dl) i a proteinemiei
(4,7-4,96 g/dl), cu rezultate aproape identice cu tratamentul medicamentos, arat o
tendin spre corectare a valorilor parametrilor investigai, care sugereaz
necesitatea prelungirii tratamentului pe durata de peste 2 luni de zile.

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BIBLIOGRAFIE

1. Sapcaliu Agripina, I. Radoi si colab., 2004 - Cercetri privind elaborarea formulei


farmaceutice i caracterizarea proprietilor fizico-chimice i microbiologice ale
produsului apiterapeutic (referat 1 la contractul 4532/2004)-susinut la data de
30.11.2004
2. Sapcaliu Agripina, I. Radoi si colab., 2006 - Testarea preclinica a produsului
obinut. (referat 3 la contractul 4532/2004)-susinut la data de 15.11.2005.2006)
3. Sapcaliu Agripina, I. Radoi si colab., 2005 - Cercetri privind posibilitile de
condiionare a produsului apiterapeutic (referat 2 la contractul 4532/2004)-
susinut la data de 28.02.2005)
4. Mihai D., 2000 - Medicina intern a animalelor, vol. 1, Edit. GEEA, Bucureti.
5. Neacsu C., 2002 - Compendiu de apiterapie. Edit Tehnica, Buc
6. Rdoi I. M. Cornila, 2001 - Principii generale de terapie alternativ i
complementar. Edit. Material didactic USAMV, Bucureti.
7. Schoen Allen M., Susan G. Wynn, 1998 - Complementary and veterinary medicine :
principles and practice. Mosby Inc. St. Louis, Missouri.
8. Willian Collinge, 1997 - Cartea Medicinei. Ghid complet de medicin
complementar, Ed. Lucman

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VRSTA I GREUTATEA CORPORAL - FACTORI DE


RISC PENTRU BOALA PARODONTAL LA CINE
AGE AND BODY WEIGHT AS RISK FACTORS
FOR PERIODONTAL DISEASE DOG

Luminia COSTINAR, Corina Mariana PASCU


U.S.A.M.V. Timioara

Periodontal disease is a common oral disease in dogs. Periodontal


disease means a series of changes that are associated with the inflammation
and loss of the deep supporting structures of the teeth. Numerous studies
demonstrated a significant correlation between greater dental calculus and
advanced age.
Periodontitis was also demonstrated to increase significantly along
with increasing age and decreasing body weight resulting in a markedly
greater incidence of disease in the older dogs, belonging to small breeds.

Boala parodontal este considerat a fi una dintre cele mai frecvente


afeciuni care apare la animalele de companie i n mod special la cine. Ea este
considerat a fi o boal inflamatorie cronic manifestat clinic prin pierderea
esuturilor de suport ale dintelui i resorbia osului alveolar. Cauza primar a bolii
este reprezentat de placa dentar.
Conform datelor recent publicate n literatura de specialitate, aproximativ
75-85% din cinii aduli sufer de diverse boli ale cavitii orale, i aproape toi
cinii n vrst de peste 5 ani au boala parodontal (1, 2, 3).

MATERIAL I METOD

A fost luat n studiu un numr de 94 de cini de rase i sexe diferite, cu


vrste cuprinse ntre 5 luni i 14 ani. Cazuistica luat n studiu a fost reprezentat
de animalele care au fost consultate n Clinica de Boli Infecioase i Serviciul de
Gard al Facultii de Medicin Veterinar Timioara i Clinica Bioveti-Farm
Timioara, n perioada ianuarie 2004 ianuarie 2006. Examinarea cavitii orale
s-a fcut pe animale contiente, atunci cnd a fost posibil, sau pe animale
anesteziate. Unii cini au prezentat diverse grade de evoluie a bolii parodontale
iar alii nu au prezentat leziuni ale parodoniului. Pentru fiecare cine s-a ntocmit
o fi stomatologic. Evaluarea stadiilor bolii parodontale s-a realizat conform
metodologiei de specialitate (1, 3, 5) i anume:
- stadiul 0 gingie sntoas i dini fr depunere de tartru;
- stadiul I (stadiul timpuriu) gingivit incipient i depozite reduse
de tartru pe o suprafa mai mic de 24% din suprafaa dentar,
halen, uoar desprindere gingival i depozite de exsudat ntre
dinte i gingie;

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- stadiul II (stadiul moderat) gingivit cronic, hemoragii gingivale la


palpaie i la sondajul cu sonda parodontal, depunere de tartru pe o
suprafa a dintelui de 24-49%, parodontit incipient, pierderea
aderenei gingivale cu formarea de pungi parodontale, mobilitatea
dentar nc nu este prezent.
- stadiul III parodontit evoluat, gingivit cronic-ulcerativ, retracie
gingival cu pn la 40% n cazul caninilor i premolarilor, acoperirea
cu tartru peste 50-74% din suprafaa dentar; apare o uoar
mobilitate dentar.
- stadiul IV (sever) parodontit avansat, cu retracie gingival care
depete 50%, descoperire radicular, halen, depozite purulente i
alimentare, pungi parodontale adnci, tartrul acoper peste 75% din
suprafaa dentar., mobilitate dentar exagerat.

REZULTATE I DISCUII

n perioada anilor 2004-2006 au fost examinai n Clinica de Boli


Infecioase i Serviciul de Gard al Facultii de Medicin Veterinar Timioara i
n Clinica Bioveti-Farm Timioara un numr de 94 de cini.
Din analiza datelor prezentate n tabelul 1 rezult c, din numrul total de
94 de cini examinai, la un numr de 58 de cini adic la 61,7% a fost
diagnosticat boala parodontal n diverse stadii de evoluie.

35
30
nr. cini examinai

25
20
15
10
5
0
0-1an 1-3 ani 3-6 ani 6-9 ani 9-14 ani
vrsta

Graficul 1 - Numrul cinilor examinai i categoriile de vrst

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Tabelul 1
Distribuia pe vrste i sexe a parodontopatiilor la cinii examinai

Cini examinai Cini cu


Categoria
Total Femele Masculi parodontopatii
de vrst
Nr. % Nr. % Total Procent
< 12 luni 8 5 62,5 3 37,5 1 12,5
1an-3 ani 25 13 52,0 12 48,0 10 40
3 6 ani 31 17 54,8 14 45,1 20 64,5
6 - 9 ani 13 9 69,2 4 30,7 11 84,6
9-14 ani 17 10 58,8 7 41,1 16 94,1
Total 94 54 57,4 40 42,5 58 61,7

Tabelul 2
Distribuia parodontopatiilor pe rase i vrste

Categoria de vrst Cini cu


Talia rasei 1-3 3-6 9-14 Total parodontopatii
< 12 luni 6-9 ani
ani ani ani Nr. Procentul
Talie mic* 4 (1)**** 7 (3) 14(12) 6 (6) 11 (11) 43 33 76,74
Talie
1 10 (6) 7 (6) 2 (1) 5 (4) 26 17 65,38
mijlocie**
Talie
*** 3 8 (1) 10 (2) 5 (4) 1 (1) 25 8 32,0
mare
Total cini
8 25 31 13 17 94 58 61,7
examinai
Legend:
*
cini de talie mic: Pudell, Pekinez, Beagle, West Highland white Terrier,
Yorkshire Terrier;
**
cini de talie mijlocie: Fox Terrier, Shar-pei, Cocker, Boxer, Pudell mijlociu, Bull
Terrier, Samoyed;
***
cini de talie mare: Ciobnesc german, Vizsla, Ciobnesc de Berna, Rotweiller,
Ciobnesc mioritic;
****
cifrele din parantez reprezint numrul de cini cu afeciuni parodontale.

Din tabelul 1 i graficele 1 i 2 se poate observa c frecvena apariiei


parodontopatiilor la cine este n strns corelaie cu vrsta. Astfel, la cinii din
categoria de vrst sub 12 luni au fost examinai 8 cini, 5 femele i 3 masculi. La
aceast categorie de vrst a fost diagnosticat doar un singur cine cu stadiul I de
boal parodontal (gingivit incipient).
La categoria de vrst cuprins ntre 1 an i 3 ani, 40% (10 cini) examinai
au prezentat leziuni ale parodoniului (tabelul 1). La aceast categorie cinii care
au prezentat afeciuni parodontale au avut vrste peste 2 ani. n aceast categorie
de vrst s-au examinat 13 femele i 12 masculi, dintre care 9 au prezentat stadiul
I de boal parodontal i un singur cine, n vrst de 3 ani, a prezentat stadiul III.

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100 94,1%
84,6%
80
64,5%
60
40%
40

20 12,5%

0
0-1 an 1-3 ani 3-6 ani 6-9 ani 9-14 ani

Graficul 2. Distribuia procentual a parodontopatiilor pe categorii de vrste

Din tabelul 1 i graficul 2 se poate constata c semnele clinice de gingivite


i parodontite pot s apar nc de la vrsta de 3 ani. Rezultate similare au fost
descrise i n literatura de specialitate (1, 2, 3).
La categoria de vrst cuprins ntre 3-6 ani s-a depistat boala parodontal
la 20 de animale, respectiv 64,5% din totalul de 31 de cini examinai (17 femele
i respectiv 14 masculi). Cinii au prezentat diverse grade de evoluie a bolii
parodontale, n sensul c: 6 cini au avut stadiul I de boal parodontal
(gingivit), 3 cinii stadiul II, 9 stadiul III, iar 2 cini au avut stadiul IV.
La categoria 6-9 ani au fost examinai 13 cini (9 femele i 4 masculi). Din
totalul acestora, 11 cini (84,6%) au prezentat leziuni parodontale n toate stadiile
de evoluie a bolii (1 cine stadiul I, 4 stadiul II, 6 stadiul III i respectiv 1 stadiul
IV).
n categoria de vrst 9-14 ani au fost examinate un numr de 17 animale,
dintre care 10 femele i 7 masculi. La aceast categorie rezultatele examinrii
clinice arat un maxim de mbolnvire, de 94,1% (16 cini). Menionm c la
aceast categorie de vrst predomin stadiile finale ale bolii parodontale, avnd
n stadiul III al bolii 4 cini, iar n stadiul IV, 11 cini.
n urma cercetrilor efectuate se constat c frecvena bolii parodontale
exprimat procentual crete progresiv odat cu vrsta, de la 12,5% la categoria de
vrst sub 1 an, la un maxim de 94,1% la categoria de vrst cuprins ntre 9 i
14 ani.
Din tabelul 2 se poate constata faptul c incidena bolii parodontale este
mult mai mare la cinii din rasele mici comparativ cu cei care aparin raselor de
talie mare.

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Astfel, la cinii de talie mic - Pekinez, Pudell, Terrier- s-a nregistrat


procentul cel mai ridicat de mbolnvire, respectiv 33 cini cu afeciuni ale
parodoniului, (76,74%) dintr-un numr total de 43 de cini examinai.
De Bowes i col. (1996) gsesc chiar c 90% din cinii care aparin raselor
mici, cu vrste peste 3 ani, prezint diverse grade de evoluie a bolii parodontale
(4).
La cinii din rasele mijlocii (Fox Terrier, Cocker, Boxer, Pudell mijlociu)
au fost examinai 26 de cini, din care 65,38% (17 cini) au prezentat
parodontopatii.
Au fost examinai i un numr de 25 de cini din rase cu talie mare. La
aceste rase s-a constatat c procentul de mbolnvire a fost de 32,0%. Aceste
rezultate concord cu cele din literatura de specialitate, i sunt reprezentate n
graficul 3.

80
70
% cini cu parodontopatii

60
50
40
30
20
10
0
rase mici rase mijlocii rase mari

Graficul 3 - Distribuia parodontopatiilor n funcie de ras

ntr-un studiu efectuat de Bolte i col. (2004) pe 1325 de cini, procentul


afeciunilor parodoniului a fost de 4,3% la categoria de vrst 6-12 luni i de
65,4% la vrsta cuprins ntre 11-14 ani (5).

CONCLUZII

Ponderea cazurilor de boal parodontal din totalul cinilor examinai n


Clinica de Boli Infecioase, Serviciul de Gard al Facultii de Medicin
Veterinar Timioara i Clinica Bioveti-Farm Timioara n perioada anilor 2004-
2006, a fost de 61,7% din numrul total de 94 al cinilor luai n studiu.

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Rezultatele noastre arat c exist o strns corelaie ntre vrsta cinelui i


boala parodontal: frecvena apariiei bolii parodontale crete progresiv, odat cu
vrsta.
Odat cu naintarea n vrst a cinelui, crete i severitatea bolii
parodontale.
Rasa are un rol major n dezvoltarea bolilor dentare, n special a bolii
parodontale, cei mai susceptibili fiind, din acest punct de vedere, cini din rasele:
Pudell, Pekinez, Terrier.
Dup vrsta de 3 ani se pot observa la nivelul parodoniului leziuni care se
pot ncadra n stadiul III i chiar n stadiul IV al bolii parodontale, manifestndu-
se prin formarea pungilor parodontale, retracie gingival i, uneori, mobilitatea
dinilor afectai.

BIBLIOGRAFIE

1. Braga Bitencourt A., Carla, Silva, Rezende, Faria, Costa, J.E., Franco Da Silva,
L.A., Farias, L.M., Carvalho de Roque, Maria Auxiliadora, 2004 - Importncia
da avaliao clinica no diagnstico de doena periodontal em ces da raa
pastor alemo, Cincia animal Brasileira, 5, 1, jan.-mar., 33-38
2. Lepine, A.J., Cox, E.R., 2001 - Nutritional Influences on Dental Health, Current
perspectives in senior Dog and Cat Nutrition, Tufts Animal Expo, 47-51
3. Harvey, C.E., O'Brien, J.A., Rossman, L.E., Stoller, N.H., 1983 - Oral, Dental,
Pharyngeal, and Gland Disorders, In: Textbook of Veterinary Internal Medicine-
Diseases of the Dog and Cat, Ettinger, S.J., second Edition, California Animal
Hospital, Los Angeles, California, p. 1126-1160
4. De Bowes, L.J., Mosier, D., Logan, E., 1996 - Association of periodontal disease
and histologic lesions in multiple organs from 45 dogs, J. Vet. Dent., 13(2)57-60
5. Bolte, S., Igna, C., Petru, H., Nicoar, F.R., Sala, A., Larisa, Schuszler, 2003 -
Aspecte epidemiologice i morfoclinice n periodontopatii la cinele din mediul
urban, Lucr. t. Timioara, vol. XXXVII, 560

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INFLUENELE DIETEI ASUPRA BOLII PARODONTALE


DE LA CINE
DIETARY INFLUENCES ON PERIODONTAL DISEASE IN DOG

Luminia COSTINAR
U.S.A.M.V. Timioara

The primary etiologic agent associated with periodontal disease is


bacterial plaque and by-products. Specific risk factors that contribute to the
severity and progression of dental diseases include: breed, age, species,
characteristics food and nutrition, imunocompetence and systemic health
(5, 7).
Food texture and composition can directly affect the oral health and
oral environment. The consistency of a diet as well as the nutritional
components can affect the rate of calculus accumulation.
The aim of the study was to determine the health condition of the
mouth cavity in dogs associated with their feeding.
Dogs fed the soft diet accumulated more plaque and developed more
gingivitis than when they were fed the hard diet.

Cauza primar a gingivitelor i parodontitelor este reprezentat de prezena


plcii bacteriene pe suprafeele dentare. Dac placa dentar nu este ndeprtat, n
aproximativ dou zile va ncepe mineralizarea acesteia, rezultnd n final tartrul
dentar. Periajul sau splarea dinilor este un mijloc important de ndeprtare a
unui mare numr de bacterii, bacterii care se gsesc att n placa dentar ct i n
tartrul dentar (3, 4, 6).
Consistena i textura dietei au un rol deosebit n acumularea depozitelor
dentare i, ca urmare, n dezvoltarea i progresia parodontopatiilor la cine.
Sntatea oral, att a cinilor ct i a pisicilor, este influenat ntr-o mare
msur de consistena alimentelor (6, 7, 8).
Boala parodontal mai are nc multe necunoscute n ceea ce privete
etiologia i evoluia, n special la animale, la care se poate considera c
cercetrile se gsesc undeva la nceput.
Acest studiu a avut drept scop elucidarea, sau completarea cunotinelor
existente asupra unui singur aspect, respectiv a relaiei dintre diet i evoluia
parodontopatiilor la cinii din oraul Timioara.

MATERIAL I METOD

A fost luat n studiu un numr de 94 de cini de rase i sexe diferite, cu


vrste cuprinse ntre 5 luni i 14 ani. Cazuistica luat n studiu a fost reprezentat
de animalele care au fost consultate n Clinica de Boli Infecioase, Serviciul de

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Gard al Facultii de Medicin Veterinar Timioara i Clinica Bioveti-Farm


Timioara n perioada ianuarie 2004 ianuarie 2006 .
Pentru fiecare pacient s-a ntocmit o fi stomatologic, pe aceast fi,
printre altele s-a nregistrat i tipul hranei. n funcie de textura hranei, cinii au
fost mprii n 3 grupe i anume:
Cini hrnii cu o mncare uscat preparat industrial, avnd o consisten
dur (crochete, biscuii).
Cini hrnii cu o mncare variat sau mixt.
Cini hrnii exclusiv cu o mncare umed, cu consisten moale (conserve
sau mncare preparat n cas resturi menajere).
Din pcate, nu a fost posibil alegerea animalelor astfel nct media de
vrst a celor 3 grupe s fie egal.
Media de vrst a grupului 1 a fost de 3,5 ani, la grupul 2 media a fost 5,3
ani, iar ultimul grup a avut o medie de vrst de 6,4 ani.
Din anamnez a reieit faptul c igiena dentar a cinilor a fost deficitar
sau chiar absent, n majoritatea cazurilor. Aceste date se bazeaz, n totalitate,
pe relatrile proprietarilor cinilor examinai.

REZULTATE I DISCUII

Din analiza datelor prezentate n tabelul 1 rezult c, din numrul total de


94 de cini examinai, 21 de cini au fost hrnii exclusiv cu hran uscat. La
acest grup de cini procentul parodontopatiilor a fost de 47,6%, respectiv 10 cini
au prezentat semne clinice de gingivite i/sau parodontite.
Tabelul 1
Distribuia parodontopatiilor la cine n funcie de tipul alimentaiei

Numrul de cini Cini cu parodontopatii


Grupul Tipul hranei
examinai Numrul Procentul
1 Hran uscat 21 10 47,6
2 Hran mixt 28 22 78,5
3 Hran umed 45 40 88,8

Un regim alimentar cu o consisten dur va duce la ndeprtarea mecanic


a plcii dentare i deci a florei bacteriene, avnd loc o curire parial a cavitii
orale. Ca urmare, se reduce severitatea bolii parodontale sau evoluia bolii poate
ncetini, dar aceast msur nu este considerat protectiv.
La cinii din grupul 2, care au fost hrnii cu mncare variat sau mixt,
procentul mbolnvirilor a fost de 78,5%, respectiv din totalul de 28 de cini
examinai, 22 de cini au prezentat leziuni ale parodoniului.
La grupul 3 de cini, care au fost hrnii cu mncare cu consisten moale
(mncare preparat n cas sau conserve) s-a constatat un procent de 88,8% a
parodontopatiilor.
Se cunoate din literatur faptul c o diet care este compus din alimente
moi va duce la formarea unui numr mai mare de plci dentare. Acest fapt se

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datoreaz, probabil, adezivitii mai mari a acestor alimente. n aceast situaie,


posibilitatea reteniei mecanice a florei bacteriene i aderarea la suprafeele
dentare este foarte mare i, ca urmare, incidena afectrii parodoniului este
ridicat (3, 4).
Aa se explic i procentul mare al parodontopatiilor ntlnit la cinii care
au fost hrnii cu mncare preparat n cas.
Alimentele moi favorizeaz ptrunderea lor n anurile gingivale i, n
consecin, are loc retenia i nmulirea florei bacteriene, n principal a florei
anaerobe, flor care este responsabil de distruciile tisulare de la nivelul
parodoniului.
Menionm c toate aceste constatri sunt n concordan cu datele
raportate i de ali autori, n literatura de specialitate (2, 6).
nc din anul 1939 Burwasser i Hill (1) au constatat faptul c o diet moale
duce la apariia unor modificri patologice la nivelul esutului gingival. De-a
lungul anilor au fost efectuate numeroase studii care susin ideea c, la cinii care
sunt hrnii cu alimente care au o consisten moale crete viteza de acumulare a
plcii dentare i a tartrului i, ca urmare, procentul afeciunilor parodontale este
mai mare. Tot la aceti cini se constat o creterea a incidenei i severitii
parodontopatiilor, n comparaie cu animalele hrnite cu alimente care au o
consisten crescut (2).

100

80

60

40

20

0
hran uscat hran mixt hran moale

cini examinai % parodontopatii

Graficul 1 - Prevalena parodontopatiilor la cini i tipul de hrnire al acestora

Acest grafic demonstreaz rolul important pe care l deine tipul dietei i


msurile de ngrijire dentar asupra sntii cavitii orale.

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Lipsa complet sau insuficienta ngrijire dentar de acas a cinelui va


permite plcii i tartrului s se formeze n mod continuu i, ca urmare, incidena
bolii parodontale va fi foarte ridicat.
n ultimii ani au aprut formule diferite de hran uscat care conin diverse
componente, cum ar fi polifosfaii, care au rolul de a preveni mineralizarea plcii
i transformarea ei n tartru dentar. Prin sechestrarea calciului n placa dentar are
loc formarea unor complexe de calciu solubile, care vor difuza n saliv i, ca
urmare, placa dentar nu va deveni tartru. Unii autori (5), susin ideea c, singura
i cea mai eficient metod de prevenire i control a bolii parodontale este
reprezentat de splarea regulat pe dini - zilnic dac este posibil iar alii (6)
recomand metode mecanice de reducere a plcii i tartrului printr-o textur dur
a dietei.

CONCLUZII

Tipul dietei este un factor de risc n dezvoltarea bolilor dentare la cine.


Frecvena apariiei parodontopatiilor este mult mai ridicat la cinii care
sunt hrnii cu o mncare umed, moale de tipul conservelor sau resturilor
menajere, dect la cei care sunt hrnii cu o mncare uscat comercial.
Ponderea cazurilor de boal parodontal din totalul cinilor examinai de
noi n perioada ianuarie 2004 - ianuarie 2006, a fost de 88,8% la cinii hrnii cu o
diet cu textur moale, 78,5% la cinii hrnii cu hran mixt, i de doar 47,6% la
cei cu o diet uscat, dur.
Consistena dietei, precum i compoziia ei, poate afecta rata acumulrii
tartrului dentar.

BIBLIOGRAFIE

1. Burwasser, P., Hill, T.J. 1939 - J. D. Res., 18:237


2. Gawor, J.P., Jodkowska, K., Kurski, G., Kurek, A., Wojtacki, M.P., 2005 - Pet
Smile Campaign in Poland-Promotion of Oral Hygiene and survey of oral cavity
in Polish pets, www.wsava 2005.com
3. Lepine, A.L., Murray, S.M., Cox, E.R., 2003 - Recent advances in Dental Health
Management 26-32
4. Harvey, C.E., Shofer, P.S., Laster, L., 1996 - J. Vet. Dent., 13:101-105
5. Tromp, J.A., van Rijn, L.J., Jansen, J. 1986 - J. Clin. Periodontol. 13:190-194
6. Wattson, A.D.J., 1994 - Australian Veterinary Journal, 313-318
7. Hoffmann, Th., Geangler, P., 1996 - Journal of Small Animal Practice, 37, 309-316
8. Rawlings J.M., Gorrel, Cecilia 1997 - J. Small Anim. Pract., 38: 147-151

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DATE PRIVIND STATUSUL SELENIULUI IN FURAJELE


DIN MOLDOVA

FODDER PLANT SELENIUM DEFICIENCY FROM MOLDOVA COUNTY

L. MIRON, Maria SERDARU, D. ACATRINEI

This paper present some preliminary data regarding the distribution


an deficency of slenium in 231 samples of different plants used like fodder
for domestic animals, from 67 localities covering 7 departmens of Moldova
county. The spectrofluorimetric examination of samples revealed a
marginal deficiency of this nonmetallic element (32,46%), a critical
deficiency (59,4 %) and a severe deficiency (3,6%) of examinated samples.

Key words: selenium status , fodder

Studiul de fa prezint date preliminare privind deficitul de seleniu din


plante utilizate ca furaj n judeele Moldovei. A fost realizata o ancheta
nutriional colectnd date privind natura furajelor utilizate pentru alimentaia
animalelor din gospodariile populaiei, randomizat, pentru a acoperi uniform
suprafaa fiecrui jude din Moldova. Prin ancheta nutriional s-au superpozat
factorii de mediu cu cei abiotici, stabilindu-se interferenele plurifactoriale ale
acestora, cum ar fi: specia cea mai atractiv de animal utilizat pentru cretere
ntr-o anumit zon geografic a rii, obinuina localnicilor de a cultiva numai
anumite specii de plante, creterea ainmalelor numai cu anumite tipuri de furaje,
furaje concentrate sau combinate furajere administrate obinuit n hrana
animalelor de curte, utilizarea mai mult sau mai puin corect a unei alimentaii
corespunztoare pentru animalele de rent. Contextul acestui studiu este mai larg,
urmrindu-se evaluarea statusului seleniului din Romnia i efectuarea unor
cartri ale zonelor pentru a cunoate limitele de variaie ale acestui element n
lanul trofic plant-animal-om.

MATERIAL I METOD

Au fost colectate 231 de probe de furaj din 67 de localiti din 7 judee ale
Moldovei, s-a realizat ancheta nutriional pentru fiecare gospodrie sau
exploataie zootehnic n parte.
Probele colectate au fost investigate prin metoda spectrofluorimetric
folosind un spectrofotometru SPEKOL echipat cu anexe pentru fluorimetrie i 2,3
diaminonaftalina ca reactiv specific.

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REZULTATE I DISCUII

In mod sintetic, tabelul 1 prezint tipurile de furaje administrate n


gospodriile populaiei sau la ferme de animale n judeele enumerate mai jos. Se
constat administrarea deopotriv a furajelor fibroase, leguminoase, concentrate,
n procente diferite n mod particular de la un jude la altul, indeosebi la
cresctorii particulari cu efective mici de animale destinate consumului sau
produciei de lapte.
Tabelul 1
Tipuri de furaje administrate

Legumi- Oleagi- Concen-


Cereale Fibroase Fin Pleav Fn
noase noase trate
Nume
Ferm

Ferm

Ferm

Ferm

Ferm

Ferm
jude

Ferm

Ferm
G.P.

G.P.

G.P.

G.P.

G.P.

G.P.

G.P.

G.P.
Bacu 5 5 1 2 2 - 1 - - - - - - 1 3 1
Iai 2 12 1 2 1 3 - 1 - 1 - - 1 2 2 11
Botoani 15 3 2 2 2 - 1 - - - - - - - 7 -
Vaslui 19 5 9 3 3 5 2 3 - - - 1 1 4 10 6
Suceava 59 2 29 - 20 2 - - - - - - 1 - 49 2
Neam 11 9 3 2 1 - - - - - 1 - 2 1 16 5
Brila 1 2 2 - - - 2 - - - - - 1 - - -
Galai 2 3 5 2 3 1 - - - - - 1 3 11 2
Buzu

Se poate observa c ponderea furajer este ocupat de cereale, n toate


judeele, att la gospodriile populaiei ct i la ferme. Urmeaz fnurile i
leguminoasele. Fibroasele, oleaginoasele i pleava sunt n ordine descresctoare
slab reprezentate.
Pentru judeul Bacu, furajele pe care le administraz proprietarii sunt :
fasole boabe, porumb boabe, gru (boabe sau tre), soia, ovz, mazre, orz.
Dintre fibroase, puini particulari administreaz lucern, majoritatea fiind axai pe
grosiere de tipul borceag, otav, fn de iarb. ntruct aceast compoziie poate
influena deficitul de seleniu, ne asteptm ca unele probe din totalul celor
analizate s confirme acest lucru.
Pentru jud. Botoani, creterea bovinelor se face preponderent pe seama
fibroaselor, constatnd un amestec cvasiomogen la cresctorii particulari de fnuri
naturale sau fn de lucern sau borceag, in combinaie cu boabele de secar,
floarea soarelui, a ovzului, grului, orzoaicei i soia. La ovine, raia este
asigurat de cca. 1 kg fn de deal, porumb uruit 1 kg, , paie de orz i sare , ori
siloz (1 kg) dac ovinele sunt crescute n ferme.

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Judeul Iai are ca particularitate amestecul grosier de strujean i lucern,


administrat alturi de lucern, flor spontan de deal iar la unele gospodrii
administreaz i borceag. Otava, soia, porumbul , orzul i grul completeaz
raiile pentru animale mari. Sunt administrate i concentrate, n amestecuri
particulare de la un cresctor la altul. Sfecla furajer este administrat la ovine,
cam 0,5 kg, alturi de porumbul boabe mcinat, cu mas verde sau vrejuri de soia.
n judeul Vaslui, cocenii de porumb per se sau tocai, fnurile de trifoi,
lucern precum i paiele de ovz sunt baza alimentaiei fibroase, borceagul fiind
rar ntlnit. Boabele de porumb, gru i secar, constituie baza raiilor la
gospodriile populaiei. Proprietarii mai fac un furaj combinat n care amestec
graminee i leguminoase uruite.
Suceava este dominat de prezena unor fnuri de bun calitate, fn de
lucern, de timotei, fn de trifoi sau fn de ierburi de munte, care asigur un
complet mineralo-vitaminic la care se adaug concentrate substaniale n raii,
comparativ cu celelalte judee. Grul, ovzul secara, fasolea i porumbul n
diferite cantiti se admnistreaz uruite. O vac primete zilnic o raie compus
din 8-10 kg fn, la care proprietarul (privat, cu 5-10 capete bovine) adaug aa
zisul ciubr compus din suculente 10-15 kg (toctur): tre, cartof fiert,
coceni tocai, dovleac, gulii, mai puin sfecl. Se adaug maxim 2 kg concentrate
(tre de porumb) la muls. Producia lactat este n medie de 10 litri de lapte.
n Neam, deasemeni, cresctorii sunt interesai de produciile ridicate de
lapte i de creterea ovinelor n condiii bune, n zonele de deal i de munte.
Pentru raia bovinelor, reeta este compus din 5+8 kg fn, 10-15 kg sfecl, 15+20
kg siloz, paie, coceni, i 2 kg de concentrate (tre gru + rot floarea
soarelui).uruiala de porumb nu prea este administrat, dect cernut (tre).
Cobornd spre Brila- Galai, constatm o cretere n raii a concentratelor
i administrarea de uruial de coceni, tiulei de porumb i lucern. Ca i boabe,
se administreaz ovz, mzriche, porumb, mazre, orz, fiind completate cu fn
de sparcet, iarb de Sudan, fn de borceag, rapi, fn de es sau fn de deal.
Deasemeni, n Galai se obinuiete administrarea de malat(praf de moar)
amestecat cu fn, coceni i paie la bovine. La oi, tot n acest areal se
administreaz fn lucern, cu porumb tiulei (cu totul), orz, gru, soia i rapi, la
care se adaug vreji (fasole, soia).
Din cele 231 de probe colectate, pn n acest moment au fost analizate 111
(48,05%). Conform NRC 1996, nivelul normal al seleniului n plante se
ncadreaz ntre 0,15-0,3 ppm. Deficiena de seleniu este marginal la nivelul 0,1-
0,15 ppm, critic n intervalul 0,1-0,01 ppm i sever la valori mai mici de
0,01 ppm.
Status-ul seleniului n probele colectate de noi este redat n valori absolute
i procentual n tabelul nr. 2. i n graficul nr. 1.

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Tabelul 2
Status-ul seleniului n probele de furaj colectate

Deficien
Nivel normal
Marginal Critic Sever
Nr probe 5 36 66 4
% din probe
4,5 32,43 59,45 3,6
analizate

Graficul deficitului de seleniu n plantele folosite ca furaj n zona


Moldovei (%)

Normal
3.6 4.5
Marginal
32.43
Critic

Sever

59.45

CONCLUZII

Patru din cele 5 probe n care seleniul a fost gasit n concentraii normale
provin din jud. Vaslui (Perieni, Vleni, Tutova, Vaslui) iar una din jud. Neam
(Praznov).
Probele cu deficit sever au fost recoltate din jud. Suceava (Rdui i aru
Dornei), jud. Bacu (Buhui) i una din jud. Iai (Ciurea).
Deficitul marginal i cel critic de seleniu sunt repartizate relativ uniform ca
dispoziie geografic.
95,5 % din probele de furaj analizate prezint un deficit de seleniu care
trebuie compensat.

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BIBLIOGRAFIE SELECTIV

1. Banuelos, G.S., D.W. Meek, 1990 - Accumulation of selenium in plants grown on


selenium-treated soil. J.Environ. Qual. 19:772-777.
2. Beath, O.A., H.F. Eppson, C.S. Gilbert, 1937 - Selenium distribution in and
seasonal variation of vegetation occurring on seleniferous soil. J. Am. Pharm.
Assoc. 26:394-398.
3. Behne, D., P. Brtter, D. Gawlik, U. Fosick, W. Wolters, 1978 - Proc. 3rd Int.
Symp. on Trace Element Metabolism in Man and Animals, M. Kirchges-sner,
Techn. Univ. Munchen, 441 447.
4. Brown, D.G., R.F. Burk, R.J. Seely, K.W. Kiker, 1972 - Int. J. Vitam. Nutr. Res., 42,
588-591.
5. Brown, R.G., P.R. Sweeney, J.C. George, D.W. Stanley, E.T. Moran, Jr., 1974 -
Poult. Sci., 53, 1235-1239.
6. Fordyce, F.M., Z. Guangdi, K. Green, Liu Xinping, 2000 - Soil, grain and water
chemistry in relation to human selenium-responsive diseases in Enshi District,
China. App. Geochem. 15:117-132.
7. Jenkins, K.J., K.A. Winter, 1973 - Can J. Anim. Sci., 53,561-567.
8. Leibetseder, J., A. Kment, M. Skalicky, 1974 - Proc. 2nd Int. Symp. On Trace
Element Metabolism in Animals, W.G. Hoekstra, J.V. Suttie, H.E. Gant-her, W.
Meri/., 1975, Am. J. vet. Res., 36, 273-281.
9. Poll, E., 1968 - Contribuia la rolul seleniului in patologia puilor de gin, Tez
doctorat, Inst. Agron. Bucureti.
10. Salaniu, V., 1970 - Carenele in seleniu la viei, miei, purcei i malaci, Tez doctorat,
Inst Agron. Cluj.
11. Spallholz, J.E., L.M. Boylan, 2002 - Amelioration of arsenic toxicity in rats given
dietary seleniumsupplements; an animal model for applied amelioration of
arsenic toxicity in Bangladesh. A research proposalto UNICEF.
12. Wang, Wuyi, Yang Linsheng, Hou Shaofan, Tan Jianan, Li Hairong, 2001 -
Prevention of endemic arsenism with selenium. Current Sci. 81:1215-1219.

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DATE PRELIMINARE PRIVIND DEFICITUL SERIC


DE SELENIU LA BOVINELE DIN MOLDOVA
PRELIMINARY DATA REGARDING SELENIUM SERICAL EFICIENCY
TO THE CATTLE IN MOLDAVIA

L. MIRON, D. ACATRINEI
U.S.A.M.V. Iai
Maria SERDARU
S.N. Inst. Pasteur S.A.

This paper continue some researches regarding the monitoring of


selenium in plant-animal-human being, coordinated by Pasteur Institute,
in an complex research Programme, started in 2005. Analysing 451
samples of bovine serum, prelevated from 80 sites from Moldova county,
along the years 2005-2006, up to 2/3 of total samples presented values
under normal limits of selenium status, so we concluded a marginal
deficiency of this oligoelement in this area.

Key words: selenium, status, cattle, Moldova

Lucrarea de fa continu unele cercetri privind monotorizarea statusului


seleniului n lanul trofic- plant-anima-om coordonate de Inst. Pasteur SA n
cadrul unui program complex de cercetare ncepnd cu anul 2005. Analiznd 451
de probe de snge de bovine recoltate din peste 80 de localiti ale Moldove in
decursul anilor 2005-2006, peste dou treimi au prezentat valori ale seleniului
seric sub limitele normale.
n organismele vii, seleniul este implicat n procese fiziologice importante,
unde substituie rolul sulfului n sistemele enzimatice i de transport, acioneaz ca
antioxidant atunci cnd intr n compoziia glutation-peroxidazei, protejnd
organismul de aciunea radicalilor liberi i influeneaz fertilitatea (6, 7).
Acolo unde solurile conin cantiti suficiente de sruri de seleniu s-a
constatat scderea incidenei cancerelor de sn i colon. Organizaia Mondial a
Sntii recomand administrarea zilnic a 80 de mg de seleniu la brbai i
60 mg la femei iar Institutul American de Cercetare a Cancerului majoreaz doza
profilactic antitumoral de seleniu la 200 mg (7).
n New Epoch Time, ediia electronic din 14 februarie 2007 se
precizeaz c la Universitatea Montpellier s-a constatat c administrarea de
seleniu previne declinul cognitiv la vrstnici. La Universitatea din Carolina de
Nord s-a descoperit c dieta cu deficit de seleniu permite creterea ratei mutaiilor
virusului gripal la oarece.
La om sunt cunoscute trei boli legate strict de deficitul de seleniu: maladia
Keshan caracterizat prin dilatarea cordului, maladia Kashin-Beck manifestat

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Universitatea de tiine Agricole i Medicin Veterinar Iai

prin osteoartropatii i o form de hipotiroidism numit Cretinism Endemic


Mixedematos (CEM) soldat cu ntrziere mintal (5).
Avnd n vedere implicaiile deficitului i excesului de seleniu a devenit
necesar identificarea surselor de seleniu, stabilirea concentraiilor de seleniu din
sol, gradul lui de ncorporare n plante, transformrile pe care le sufer n lanul
trofic plant-animal-om i msurile care trebuiesc luate pentru a controla eficient
deficitul de seleniu.
Astfel a aprut necesitatea realizrii unui studiu complex iniiat de Institutul
Pasteur n parteneriat cu cele patru faculti de medicin veterinar din ar,
proiect care i-a propus monitorizarea statusului seleniului i cartarea acestui
oligoelement n ntreaga Romnie, ambele, utile pentru asigurarea sntii
omului i animalelor.

MATERIAL I METOD

Pentru evaluarea statusului seleniului la animalele din zona Moldovei s-au


recoltat 545 de probe de snge, n principal de la bovine (535), din care pn n
prezent au fost procesate 451. Determinarea seleniului seric s-a realizat prin
metoda spectrofotometric, folosind spectrofotometrul SPEKOL iar ca reactiv 2,3
diaminonaftalina.
Limitele de referin privind concentraia seleniului seric dup care s-a fcut
interpretarea rezultatelor sunt preluate din literatura de specialitate i se regsesc
n tabelul 1.
Tabelul 1
Limitele de variaie ale concentraiei serice de seleniu la animale

Specificare Nivel adecvat Deficien (ppm)


(ppm) Marginal Critic Sever
Bovine 0.040-0,1 0.010-0,040 0,005-0,010 Sub 0,005
Ovine 0.100-0.200 0.050-0.100 0.010-0.050 Sub 0.010
Suine 0.140-0.300 0.060-0.140 0.005-0.060 -

REZULTATE I DISCUII

Din 451 de probe analizate, la 313 s-a nregistrat deficien marginal de


seleniu (69.4%), la 7 probe deficien critic (1.5%), o singur prob s-a situat sub
pragul deficienei severe (0,2%) iar nivelul adecvat a fost atins la 130 de probe
(28.8%) (graficul 1).
Deficitul seleniului seric este neuniform sub raport geografic, deficiena
marginal fiind prezent n jumtate din probe n judeele Bacu i Neam, peste
dou treimi din probele provenite din judeele Botoani i Iai, i peste trei sferturi
din probele din Suceava, Galai i Brila (tabelul 2).

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Graficul concentraiei seleniului seric la animale n zona Moldovei

Deficiena marginal
Nivel adecvat
Deficien critic
Deficien sever
1.5
0.2
28.8

69.4

Graficul 1

Nivelul deficienei critice a fost atins doar n 7 probe iar cel al deficienei
severe ntr-o singur prob, toate provenite din judeul Suceava (tabelul 2).
Tabelul 2
Tabel centralizator pe judee privind concentraia seric de seleniu

Nivel adecvat Deficien (ppm)


Specificare (ppm) Marginal Critic Sever
Nr. % Nr. % Nr. % Nr. %
Jud. Suceava 14 10,3 114 83,8 7 5,15 1 0,75
Jud. Neam 9 47,36 10 52,34 0 - 0 -
Jud. Iai 6 20,69 23 79,31 0 - 0 -
Jud. Bacu 53 45,3 64 54,7 0 - 0 -
Jud. Vaslui 45 37,7 76 62,3 0 - 0 -
Jud. Galai 2 15,38 11 84,62 0 - 0 -
Jud. Brila 1 6,25 15 93,75 0 - 0 -

n judeul Suceava, deficiena critic a fost constatat la probe provenind


din localitile Neagra, Breaza, Gura Putnei, Flticeni i Voitinel. Din curtea unui
fermier din Voitinel s-au recoltat o prob cu deficit critic i unica proba cu deficit
sever din Moldova.
n judeul Iai, cele ase probe cu nivel normal al concentraiei seleniului au
fost recoltate dintr-o singur localitate, Poieni, n celelalte localiti luate n studiu
nregistrndu-se deficit marginal.

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n judeul Vaslui unele localiti au furnizat probe n care seleniul avea


nivel normal n majoritatea cazurilor (Vaslui, Tutova, Soleti) sau n care deficitul
marginal era preponderent (Chilia, Laza, Brlad, Perieni). Din Brlad provine o
prob n care seleniul era la un nivel mai mare dect cel normal, probabil ca
urmare a unei administrri de seleniu n scop profilactic sau curativ.
La Galai i Brila predomin deficitul de seleniu iar n judeul Neam n
fiecare localitate se ntlnesc valori ale deficitului marginal i ale nivelului normal
n proporii aproape egale.

CONCLUZII

Deficitul de seleniu este ntlnit la peste dou treimi din probele de snge
recoltate din judeele Moldovei.
Judeele Suceava, Brila, Galai i Iai sunt cele mai afectate de deficitul de
seleniu.
n partea de nord-est a judeului Vaslui sunt mai multe probe cu coninut
normal de seleniu dect n sud i sud-vest.

BIBLIOGRAFIE

1. Fordyce, F.M., Z. Guangdi, K. Green and Liu Xinping, 2000 - Soil, grain and water
chemistry in relation to human selenium-responsive diseases in Enshi District,
China. App. Geochem. 15:117-132.
2. Golubkina, N.A., G.V. Alfthan, 1999 - The human selenium status in 27 regions of
Russia. J. Trace Elem.Med. Biol. 13: 15-20.
3. Johnson, C.M., 1975 - Selenium in soils and plants: contrasts in conditions providing
safe but adequate amountsof selenium in the food chain. In: Trace Elements in
Soil-Plant-Animal Systems. D.J.D. Nicholas and A.R.Egan, Eds. Academic
Press, NY, pp. 165-180.
4. Spallholz, J.E., L.M. Boylan, 2002 - Amelioration of arsenic toxicity in rats given
dietary selenium supplements; an animal model for applied amelioration of
arsenic toxicity in Bangladesh. A research proposalto UNICEF.
5. http://autoimmunedisease.suite101.com/article.cfm/selenium_deficiency
6. http://www.scienceagogo.com/news/20010511055027data_trunc_sys.shtml
7. http://www.ingentaconnect.com/search/article?title=Selenium+deficiency&title_type=t
ka&year_from=1998&year_to=2007&database=1&pageSize=20&index=1

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TESTAREA EFECTULUI ANTIEIMERIAN AL UNOR


EXTRACTE DE PLANTE N EIMERIOZA
EXPERIMENTAL A PUILOR DE PREPELI
THE ANTIEIMERIAN EFFECTS OF PLANT EXTRACTS
IN QUAIL EXPERIMENTAL EIMERIOSIS

H. BACIU, V. COZMA,
Maria TRUTA, M. CERNEA
U.S.A.M.V. Cluj-Napoca

The intensiv breeding of quails knows large development


perspectives. The japonese quail Coturnix coturnix japonica,the smallest
tamed avian specie is used not only for egg production but also for meat
production. Fast growing ritm ,early sexual maturity ,short period between
generation,great egg production small fadder needs, and small space
comparativ to other bird species exploited for the for the same purpose
,those are the main caracteristics of this avian specie.
This study is the gathering of data regarding the evolution and
pathogeny of eimeriosis in quail ,as well as the establishment of some
phitoterapeutical programs in order to reduce as much as posible the
incidence of this deseases in prepelite breeding compexes.

MATERIALE I METODE
Cercetrile privind testarea efectului extractelor alcoolice din afin i
schinduf n eimerioza experimental a puilor de prepeli au fost realizare n
perioada 6 februarie 10 martie 2006, n cadrul disciplinei de Boli Parazitale a
Facultii de Medicin Veterinar, Cluj-Napoca.
Puii de prepeli luati n studiu, au fost lotizati astfel:
- Lotul I (10 pui de prepeli) - infectai cu oochisturi evoluate de eimeria
i tratai cu extract de schinduf;
- Lotul II (10 pui prepeli) - infectai cu oochisturi evoluate de eimeria
i tratai cu extract de afin;
- Lotul III (10 pui prepeli) - infectai cu oochisturi evoluate de eimeria
si netratai;
- Lotul IV (10 pui prepelita) - neinfectai i netratai;
Specificare: primele trei loturi au fost infectate doua zile consecutiv cu cte
5000 oochisturi, iar fitoterapia s-a realizat conform dozajului prescris de
producator.

Prezentarea produselor testate


Trigonella foenum-graecum (Schinduf)
Este o plant anual, ierboas, care crete spontan n culturile de cereale,
este erect, nalt de 30-60-70 cm, neramificat sau ramificat la partea
superioar i are rdcina pivotant. Tulpina este cilindric, fistuloas i glabr,

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Universitatea de tiine Agricole i Medicin Veterinar Iai

frunzele sunt lung peiolate (1-2 cm), cu stipele lanceolate, foliole oborate, uor
alungite i dinate glabre pe faa superioar i slab proase n cea inferioar.
Florile sunt grupate cte 1-2 la subsuoara frunzelor superioare, lungi de 12-18
mm, de tip papilionat, de culoare glbuie sau slab liliachie. Fructul este o pstaie
alungit dreapt sau uor curbat, lung de 8-10 cm i lat de 0,5-1 cm, ce
conine 10-20 semine comprimate, prismatice, de culoare glbuie, cu miros de
cumarin (Munteanu, 1996).
Seminele de schinduf (Trigonellae semen) sunt cunoscute nc din
antichitate de ctre medicii greci i arabi, care le acordau o deosebit atenie i le
cultivau pentru creterea ponderal n anemie, convalescen i tuberculoz.
Aceste indicaii se menin i n zilele noastre, n plus fiind folosite ca
antihelmintice, depurative, n diateza antritic i n tratamentul pelagrei.
Cercetrile recente au artat efecte favorabile i n diabet, gua exoftalmic,
tulburri digestive, nevroze, subnutriie i anorexie la copii (El Shabrawy i Nada,
1996).
Dintre substanele chimice puse n eviden pn n prezent, se remarc
proteinele, uleiul gras, mucilagiile, saponozidele steroidice, taninurile, cumarinele
i substanele cu caracter alcaloidic (numite trigoneline).
Nu se poate preciza care dintre substanele chimice coninute confer
produsului Trigonellae semen proprietile menionate anterior, ceea ce face ca,
n fitoterapie, s se foloseasc seminele integrale sau macerate apoase. Este de
menionat faptul c nici produsul vegetal medicinal i maceratele apoase nu sunt
standardizate n principii active. Nu sunt cunoscute forme farmaceutice cu acest
produs sau cu extracte ale acestuia (Franz, 1996). n Romnia, asupra acestei
plante nu a fost efectuat nc un studiu farmaceutic sistematic.

Vaccinum Myrtillus L. (Afinul)


Aparine Fam. Ericaceae i este cel mai raspsndit arbust din zona alpin a
rii noastre, formnd aa numitele afiniuri pduri miniaturale de aproximativ 50
cm. nalime, formate din arbuti ramificati,cu ramurile permanent verzi, cu frunze
alterne de, aproximativ 3cm. lungime i 2 cm. laime, scurt peiolate, de form
oval-eliptic, cu varful ascuit, glabre i cu marginea fin dinat.
Planta nflorete din mai pn la sfarsitul lunii iunie i are flori mici, de
culoare roz, solitare, actinomorfe pentamere, cu caliciul concrescut, cu ovarul de
form globuloas-urceolat.
Fructul matur este o baca zemoasa de culoare albastrui-negricioasa,
brumat de 6-10 mm diametru i are un suc violaceu cu gust placut, dulceag-
acrior.
n afara afinului spontan exist aproximativ 200 specii, din care n Europa
s-au identificat numai 4. ncepnd din anul 1925 s-a introdus afinul de cultur, cu
tufa nalt, original din America de Nord.
O serie de glicozide ale cianidolului, delfinidolului, malvidolului i
petunidolului dau culoarea specific a fructelor, care sunt bogate n: zaharuri 7-
13%; substante pectice 0.350-0.490%; tanini 0.300-0.430%; vitamina C 12-20
mg%; vitamina A 280 U.I.; PP 0.2 mg.%; B1 0.02 mg.%; B2 0.02 mg.%; E, acizi
organici, etc. De asemenea conine cantiti mari de sruri minerale, dintre care
amintim: K 50 mg.%; Ca 10 mg.%; P 8 mg.%; S 8 mg.%; Mg. 6 mg.%; Cl. 5
mg.%; Mn. 3 mg.%; Fe. 1 mg.%.

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n scopuri medicinale se utilizeaz fructele(Fructus myrtilli) i frunzele


(Folium myrtilli).
Frunzele se recolteaz vara i se usuc la umbr, iar fructele se recoltez
dup ce ajung la maturitate iunie-august i se usuc la temperaturi de 60-70C.
n frunze se gasesc derivati flavonici; ele mai contin timina, riboflavina,
vaccinina, arbutina, ericolina, o serie de acizi ( miristic, palmitic, chininic) alcooli
cerilic, miricilic precum si o glicozida a acidului galic denumita neomirtilin care
ar fi responsabil de aciunea hipoglicemiant, iar fructele conin tanini, derivati
antocianici (mirtilina a si b) 3-glucozida i respectiv 3- galactozid delfinolului,
care dau coloraia lor specific; mai conin vitamina C i complexul B, provitamina
A, principii bacteriostatice sau bactericide nc neizolate.
Proprietaile astringente i antiseptice (antibiotice) ale plantei sunt
importante, este ocrotitoarea pereilor vasculari, antisclerotica, de asemenea are
i aciune hipoglicemiant (folosit n madicina noastr popular). Frunzele intr
n compoziia ceaiurilor antidiabetice. Se foloseste ceaiul (1-2 g%) sau decoctul
(0.5-3 g%) de frunze de afin n combaterea tuturor formelor de diaree, dizenterie
cronic, ameliorand procesele de fermentaie i putrefacie intestinal.
Fructele sunt ntrebuinate n unele produse de cofetrie (gem, dulcea,
etc) afinata, precum i ca antidiareice, antidizenterice, antitifice, astringente,
stomahice, n stimularea acuitatii vizuale a aviatorilor, conducatorilor auto, ele pot
fi folosite cu bune rezultate n: hemoragii prin fragilitate capilar, retinopatii,
insuficiena biliar, azotemie i altele.
Prin vitamina P i prin aciunea trofic antiaterogen, protejeaza pereii
vaselor sanguine. De asemenea au rol coronaro-dilatator nlesnind reglarea
cardiovascular.
Rezultatele bune obinute n tratamentul cu fructe de afin se datoresc unui
comlex format din trei factori curativi: unul astringent, unul antiseptic i al treilea
absorbant. Bolul fecal devine usor acid, disparand mirosul neplacut. Antocianii
patruni n epiteliul mucoasei intestinale formnd o pelicul protectoare gri-negr,
care diminu inflamaia locala i secreia, eliminnd i stresul mecanic. La
aceasta se mai adaug efectul de inhibiie al perisaltismului intestinal exagerat
(amplitudine, tonus, frecven).

Obiective urmrite
1. Sporul n greutate i rata de conversie a hranei la loturile experimentale
2. Dinamica coproeliminrilor oochisturilor i scorul fecalelor la loturile
experimentale
3. Indicele eimeriostatic i procentul performanei eimeriostatice

REZULTATE I DISCUII

n urma cercetrilor efectuate pentru stabilirea efectului eimeriostatic,


rezultatele obinute folosind extractele de Trigonella semen i Vaccinum
myrtillus se evideniaz prin:
Sporul n greutate i rata de conversie a hranei la loturile
experimentale

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Datele referitoare privind diferenele sporului n greutate la loturile


experimentale sunt redate n Tabelul 1i n Tabelul 2.
Analiznd datele obinute s-a observat c sporul n greutate la lotul martor
negativ a fost 310 g , superior celui tratat cu Trigonella i Vaccinum la care
sporul a fost de 255 g, respectiv 265 g dar media lor a fost net superioar lotului
martor pozitiv care a avut valoarea de 225 g. n cazul lotului martor pozitiv, se
obsrv c pentru a produce un kg spor n greutate sunt necesare 3 kg furaj. n
acest sens, este cunoscut faptul c eimerioza puilor de prepeli determin
nerealizarea sporului de greutate i ntrzieri n cretere i maturizare sexual,
producnd pagube economice importante.
Tabelul 2
Conversia hranei la loturile experimentale

Spor Perioada Consum furaje Conversia hranei


Lot
(kg/lot) (zile) (kg) (kg furaj/kg spor)
I
0,255 17 0,638 2,50
Trigonella
II
0,265 17 0,662 2,49
Vaccinum
III
0,225 17 0,675 3,0
"M+"
IV
0,310 17 0,620 2,0
"M-"

Dinamica coproeliminrilor oochisturilor i scorul fecalelor la


loturile experimentale
Urmrind dinamica coproeliminrilor oochisturilor de Eimeria spp., s-a
observat c la 4 zile post infectant numrul oochisturilor eliminate a fost foarte
mare (cuprins ntre 50000 - 82000 OPG la M+). n urmtoarea perioad a
experimentului s-a observat o tendin accentuat de scdere a oochisturilor
coproeliminate, n cazul tuturor loturilor, ajungnd la limita de 400 la lotul tratat
cu Vaccinum, respectiv 1400 la lotul tratat cu Trigonella.
Extractele de Trigonella foenum-graecum i Vaccinum administrate pe tot
parcursul experimentului au acionat ca: protectoare ale mucoasei digestive;
antiinflamator; imunostimulator, iar prin alcaloizii coninui (trigonelin) ar putea
inhiba dezvoltarea i multiplicarea formelor schizogonice i gametogonice ale
parazitului.

Indicele eimeriostatic i procentul performanei eimeriostatice


Fitoterapia folosind extractele de Trigonella si de Vaccinum au pus n
evidenta indici eimeriostatici superiori martorului pozitiv (IC=346,5 respectiv
IC=356,5 comparativ cu 297,7), iar procentul performantei eimeriostatice a avut
valori apropiate la ambele loturi tratate cu valori de 85,6% Trigonella si 88,1%
Vaccinum .

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CONCLUZII

Cercetrile efectuate n perioada 6.02.2006 10.03.2006, n direcia


stabilirii efectului de stimulare a cresterii i antieimerian al extractelor de
Trigonella semen si Vaccinum myrtillus, n eimerioza experimental la puii de
prepelit, au relevat urmtoarele:
1. Cel mai mare spor n greutate a fost nregistrat la lotul tratat cu Vaccinum
cu o medie/pui de 26,5g faa de lotul martor pozitiv la care am nregistrat valori
de 22,5g.
2. Rata de conversie a hranei la loturile tratate a avut valoari apropiate, 2,50
respectiv 2,49 kg furaj/kg spor ,comparativ cu M+ care a consumat 3 kg furaj/kg
spor.
3. Loturile tratate au avut media coproeliminrilor de oochisturi de
Eimeria spp inferioara lotului M+
4. Indice eimeriostatic si procentul performantei eimeriostatice a avut valori
apropiate la cele doua loturi tratate.

BIBLIOGRAFIE

1. El Shabrawy, O.A.; Nada, S.A., 1996 Biological evaluation of multicomponent tea


used as hypoglycemic in rats. Fitoterapia, 67(2): 99-102.
2. Franz, G., 1996 Planta Medica, 14.
3. Munteanu, L.S., 1996 Cultura plantelor medicinale i aromatice. Edit. Dacia, Cluj-
Napoca.
4. uteu, I. Cozma, V., 2004 Bolile parazitare la animalele domestice. Edit. CERES,
Cluj-Napoca.
5. Williams, W., 1999 - A compartmentalized model for the estimation of the cost of
coccidiosis to the worlds poultry production industry. International Journal for
Parasitology, 1209-1229.

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RELATIONS AMONG WEIGHT AND NUMBER


OF EGGS LAID BY DERMACENTOR MARGINATUS
AND HAEMAPHYSALIS PUNCTATA AND SOME
ENVIRONMENT FACTORS
Lidia CHIIMIA*
U.S.A.M.V. Timioara

The eggs weight, laid by both species, had a high variability, thus:
eggs of Dermacentor marginatus were weighing between 0.119 and 0.524 g
with a mean value of 0.3523.816x10-2 g, and the Haemaphysalis punctata
was weighing 0.047-0.253 g with a mean value of 0.1711.806x10-2.
In natural conditions, D. marginatus females laid between 2290 and
9606 eggs with a mean of 6274.33624, and the H. punctata laid between
947 and 5900 eggs with a mean of 3550.33466.
There are correlations between the number of eggs and the
maximum temperature (r = +0.957 at p<0.001) and minimum temperature
(r = 0.797 at p<0.001).
It was found a significant correlation between the number of eggs
and the mean temperature at soil level (r = +0.939 at p<0.001), maximum
temperature at soil level (r = +0.827 at p<0.001), minimum temperature at
soil level (r = +0.793 at p<0.001), and the difference between maximum
and minimum temperature at soil level (r = +0.621 at p<0.001). There are
significant correlations between the number of eggs laid by the ixodid
females and the sunlight length (r = +0.473 at p<0.001). During the study
period it was observed that females begin to lay eggs in the evening time.
In this case it was found that there are negative correlation between
the number of eggs laid and relative humidity (r = -0.516 at p<0.001) and
rainfall (r = -0.276 at p<0.12).
It was observed that the interaction effect with some climatic factors
(for example, temperature and humidity) influence differantly and
significantly the number and eggs weight.

The ixodid ticks have a cosmopolitan and irregular spread. There are areas
where ticks presence is constant among very important populations, as well as
areas where the climatics and/or biocenotics condition permit only a sporadic and
inconstant survival [5].
The adaptation of a certain tick species to a particular microclimat depends
on its evolutive history and on the one of its host. It is in relation with genetic
factors which interact with the microhabitat specific features. The wide
distribution of tick species is based on different structural, physiological, and

*
Corresponding author: Tel. +40256277140, Fax: +40256277118
E-mail address: lidychitimia@yahoo.com

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behavioural characteristics, which is up to ticks euryoecious species on


excellence [5]. The effective presence in favorable biotopes depends on the
regional climatic fluctuations and the hosts availability in past and in present [4].
The temperature and relative humidity are the principal factors which determine
the main aspects of ticks life cycle. The mortality of ticks is the result of a
complex interaction between temperature, relative humidity, rainfall and
evaporation [8]. The geographic distribution of tick species, and indirectly of tick-
borne diseases, is limited by climatic factors. The range of any tick species is not
static, and can retreat or advance with the humidity and drought cycles [9].

MATERIALS AND METHODS

The tick females were collected in April from naturally infected sheep. After
identification according to morphological features using the determination keys of
Feider [10] and Babos [1], it was established that the species are Dermacentor
marginatus and Haemaphysalis punctata. The samples of engorged females were
chosen through drawing of lots from all the collected ticks; there were selected 12
females. Females were weighed with the analytic balance and were put in test
tubes prepared according to Metianu method. Thus, glass test tubes of 18 cm
high were used. Each test tube was half filled with water then two layers of
water cotton-logged (about 5 cm high) were placed in each tube. The test tubes
were closed with wrapped cotton plugs. During the entire experimental period the
test tubes were kept outside under the direct influence of climatic factors. The test
tubes were placed on the ground, under different bushes so that they werent
exposed to direct sunlight. Females were kept in individual test tubes in order to
determine biological characteristics.
The environment factors considered to have influence on ixodide ticks
biology were: minimum, maximum and mean air and soil temperature, rainfall,
relative humidity and sunlight duration. Meteor data were obtained from Banat-
Crisana Regional Meteorological Center from Timisoara. During the study there
were used daily values of these factors and mean value, standard deviation and
variance of these factors.
Data were processed with Microsoft Excel and statistical analysis. The
statistical analysis was carried out using SPSS 7.5 application to obtain
correlations, t test and ANOVA in order to determine, environment factors effects
on some biological parameters of these species.

RESULTS AND DISSCUTIONS

The eggs weight laid by both species was very different, thus: eggs of
Dermacentor marginatus was weighing between 0.119 and 0.524 g with a mean
value of 0.3523.816x10-2 g, and the Haemaphysalis punctata was weighing
0.047-0.253 g with a mean value of 0.1711.806x10-2. The egg mass weight of D.
marginatus taken into study by Ionita (2003) varied between 1.123 and 0.566 g

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with a mean of 0.326. It can be observed that there were very many differences
between the eggs weight of the females exposed to direct influence of climatic
factors and the ones kept in laboratory conditions.
The number of eggs laid by each female depends on the female weight and
on the environmental factors. In natural conditions, D. marginatus females laid
between 2290 and 9606 eggs with a mean of 6274.33624, and the H. punctata
laid between 947 and 5900 eggs with a mean of 3550.33466.
The number of eggs laid by each ixodid female depends on females weight,
field conditions, and the tick species (t = 3.494 at P0.002).
Tabel 1
Number of eggs value and variability depends by tick species

Std. Error of Std.


Tick species N Mean Minimum Maximum
Mean Deviation
Dermacentor
12 6274.3333 624.4584 2290.00 9606.00 2163.1873
marginatus
Haemaphysalis
12 3550.3333 466.7711 947.00 5900.00 1616.9425
punctata

Dermacentor reticulatus females laid 5000-7000 eggs [3]. In laboratory


conditions Boophilus annulatus laid 2004 342 [16], 1982); 3877 [7]; 79872064
eggs [14]. Dermacentor reticulatus and Rhipicephalus sanguineus females laid
about 4000 eggs [5]. In laboratory conditions D. marginatus females laid 2365-
10115 eggs [13].
In the following statistical analysis it was observed that between the
number of eggs and air temperature there are positive correlations: between the
number of eggs and maximum temperature (r = +0.957 at P<0.001) and minimum
temperature (r = 0.797 at P<0.001).
In the model where the mean of maximum air temperature and the mean of
rainfall (P<0.001) was introduced, a significant association between simultaneous
action of both factors and the number of eggs laid by females (P<0.014, R2 =
0.583) was observed during oviposition.
There was found a significant correlation between the number of eggs and
mean temperature (r = +0.939 at P<0.001), maximum temperature (r = +0.827 at
P<0.001) and minimum temperature at soil level (r = +0.793 at P<0.001), and the
difference between the maximum and minimum value of temperature at soil level
(r = +0.621 at P<0.001). The difference between the number of eggs laid by the
females of both species were not strongly significant (F = 12.20 la p<0.02).
Introducing simultaneously the soil temperature and rainfall quantities we can
obtain a statistic model where it can be noticed the influence of these factors on
the number of laid eggs. It can be seen that the simultaneous action of temperature
and rainfall on the number of laid eggs is not representative and significant.

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Taken into consideration how the number of eggs are correlated with air
and soil temperature we can certainly assert that the number of eggs is influenced
more by temperature of the soil level then by air temperature. Moreover, the
number of eggs is correlated with all values of temperature at soil level. This data
come to confirme what other authors said [2, 4] that temperature is a dynamic and
an essential factor for oogenesis and survival.
The development of many tick species is also influenced by humidity and
can be stopped when ambient humidity conditions are unfavorable [6]. In this case
it was found that there is a negative correlation between the number of eggs laid
by females and relative humidity (r = -0.516 at P<0.001) and rainfall (r = -0.276
at P<0.12). The relative humidity is always necessary to ensure the eggs
development and survival. The relative humidity of biotop needed by a certain
species is comprised between some specific limits of the respective species.
Nevertheless, when it is studied the effect of environmental factors on some
biological parameters is taken into consideration more the cumulative effect of
temperature and humidity.
Thus, the number of eggs produced by engorged I. rubicundus females
varied from 2045.7 (temperature 10oC and relative humidity 93%) to 3777.7
(temperature 20oC and relative humidity 93%). The number of eggs laid by a
female from this species were 2700 at 25-35oC, but it decreased significantly at
20oC (about 2300 eggs/female), at 15oC (about 1800 eggs/female) and at 40oC
(about 30 eggs/female) [17]. Humidity factors can also influence the number of
laid eggs, the number of ovipositing females, the duration of oviposition or the
moulting of the developmental stages of ixodid ticks [11, 12].
When humidity is low and the ticks continuously lose water this stage is
known as a critical equilibrium of humidity. This humidity threshold varies
between 75 and 94% relative humidity for the majority of tick species and can
differ also in different developmental stages or sexes of the same species [15].
It was observed that the number of eggs is not influenced by environmental
factors oscillations in preoviposition period. On the other hand, neither between
the environmental factors variations during oviposition and the number of eggs
laid by the ixodid tick females were individual associations obtained but only
associations between the number of laid eggs and the concomitent action of some
factors (r = +0.473 at P<0.001).
There are significant correlations between the number of eggs laid by the
ixodid females and the sunlight length (r = +0.473 at p<0.001).
During the experiment it was observed that females begin to lay eggs
during the night. The tubes, in which the ticks were placed, were examinated
twice a day (at 9 a.m. and 5 p.m.) and it was observed that neither female began to
lay eggs in day time. The ixodid oviposition takes place mainly in scotophase
time [5]. At the beginning of the ovipositon period the rythm of laying eggs is
more intense. Iwuala and Okpala [14] found that 11% of eggs were laid during the

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day, and 89% during the night, the eggs laying being maximum at dawn and dusk.
It were found similar results for A. variegatum as well. This observation is in
contradiction to that reported by Outhelli et al. [16], according to them the
engorged females exposed in laboratory at different light and darkness periods
dont influence the number of eggs.
Numerous reports have described the influence of solar radiation,
temperature and humidity on the biological cycle of ticks. Overall, the most
important factor seems to be the photoperiod, which has an impact on the
mechanisms regulating diapause in ticks and the length of their development
cycle. There are two basic types of responses to photoperiod expressed by most
ticks: 1) long-day (LD) responses, which initiate active development; and 2)
short-day (SD) responses, in which development is accelerated when perceived by
the ticks [6]. In addition, a more complex, two-step photoperiodic response is also
known in certain ticks; however, in this case, development proceeds only after
certain increases or decreases in day length. The LD or SD reaction and the one-
or two-step reactions vary in different tick species, developmental stages or
populations. Oogenesis, for instance, is regulated by the SD reaction in I. ricinus,
D. marginatus, D. reticulatus sau Ornithodoros gurneyi. In contrast, it is
regulated by the LD reaction in Hyalomma anatolicum or Argas arboreus,. Larval
development is accelerated by the LD period in I. ricinus, I. persulcatus and I.
trianguliceps. This two-step photoperiodic reaction also occurs in the
development of nymphal I. ricinus [6]. The development of this species is strongly
influenced by photoperiod rather than by temperature, humidity and food
quantity, which is considered to be of secondary importance. The minimal
induction of daily photoperiod ranges from about 14 hours light and 10 hours
darkness in connection with a minimal light intensity of 15 lux. Larvae can be
adapted partly to full darkness in the laboratory, but nymphs are unable to
metamorphose under a shorter light exposure [6].
The correlation between eggs weight and environmental factors taken into
study have the same values with the correlations between the number of eggs and
environmental factors. The correlations between the number of eggs and
temperature were observed by other authors as well [16, 17] and in these cases the
ticks were kept at certain values of temperature and relative humidity in
laboratory condition.
Regarding eggs weight it was not found an association between this
variable and the environment factors variations during preoviposition period and
we can say, thus, that environmental factors oscillation from this period dont
influence the eggs weight. But during the oviposition period the eggs weight is
dependent on some environmental factors variations.
In the model where the mean values of mean temperature of air and rainfall
(P<0.001) were introduced, it was observed an association between the interaction
of the two factors and eggs weight (P<0.001, R2 = 0.600). As well, an association

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(P<0.05, R2 = 0.529) between eggs weight and concomitant action of mean values
of maximum air temperature and the mean values of rainfall was observed.
The model where the mean values of the mean soil temperature and the
mean values of rainfall (P<0.001) were introduced, it was observed an association
between these factors interaction and eggs weight at P<0.05 and R2 = 0.521. The
same type of association was found when the variables were the mean values of
sunlight length and the mean values of rainfall.
Between the mean values of maximum soil temperature and the mean
values of rainfall and eggs weight introduced in the model (P<0.001) were
recorded associations (P<0.07).
In the model where the difference values of maximum and minimum soil
temperature and the mean values of rainfall were introduced (P<0.001), it were
observed associations between concomitant action of these factors and the eggs
weight (P<0.07, R2 = 0.512).
So, for the development of ixodid ticks life cycle, the values of
environment factors from microhabitat are more important than those from the
macrohabitat. The microhabitat can be defined as the smallest part of the habitat,
which is the actual living environment of ticks and other small organisms.
Microhabitat size varies in relation to the size of the organism, its ecological
requirements and its mobility in the terrain.
The obtained data in the following statistical analysis show us very clearly
that the effect of environmental factors interaction (temperature and humidity)
influence differently some biological parameters of ticks.

CONCLUSIONS

Between the number of eggs, eggs weight and sunlight length were found
significant correlations (P<0.001), correlations that were not recorded in special
literature.
The eggs weight and the number of laid eggs by both studied female
species were influenced by tick species and the environmental factors.
It was observed that, as well between eggs weight as between the number
of eggs and maximum and minimum values of air temperature and maximum,
minimum and mean values of temperature at soil level and the difference between
maximum and minimum temperature at soil level were significant positive
correlations (P<0.001). Due to these correlations we can assert that the values of
temperature at soil level influence more the eggs weight varieties and the number
of eggs in the ixodid ticks.
The correlations existent between the two biological parameters and
relative humidity and rainfall were negative (P<0.001and respectively, P<0.12).
All females began to lay eggs during the night.

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It was observed that the cumulative effect of the action of some


environmental factors (temperature and humidity) influence differently and
significantly some biological parameters of ticks, such as the number of eggs and
eggs weight.

BIBLIOGRAFIE

1. Babos S., 1964 - Die Zeckenfauna Mitteleuropas. Akademiai Kiado, Budapest.


re
2. Bourdeau P., 1993 - Les tiques dimportance vtrinaire et mdicale. 1 partie:
Principales caractristiques morphologiques et biologiques et leurs
consquences. Point Vet., 25, 151, 13-26.
3. Cernianu C.C., 1957 - Piroplasme i piroplasmoze. Vol. I, Ed. Academiei Republicii
Populare Romne.
4. Chartier C., Itard J., Morel P.C., Troncy P.M., 2000 - Prcis de Parasitologie
Vtrinaire Tropicale. Ed. TEC&DOC, Paris.
5. Cosoroab I., 2000 - Parazitologie veterinar. Acarioze i entomoze. Ed. Mirton,
Timioara.
6. Daniel M., Dusbbek F., 1994 - Micrometeorological and microhabitat factors
affecting maintenance and dissemination of tick-borne diseases in the
environment. n Sonenshine, D.E., Mather, T.N., 1994. Ecological dynamics of
tick-borne zoonoses. Oxford University Press.
7. Davey R.B., Garza J.Jr., Thompson G.D., 1982 - Seasonal observations on the
development and ovipositional capability of Boophilus annulatus and B.
microplus reared on bovines. J. Med. Entomol., 19, 1, 24-28.
8. Drummond R.O., Whetstone T.M., 1970 - Oviposition of the Gulf Coast tick. J.
Economy Entomol., 62, 934-936.
9. Estrada-Pea A., 2001 - Forecasting habitat suitability for ticks and prevention of
tick-borne diseases. Vet. Parasitol., 98, 111-132.
10. Feider Z., 1965 - Fauna R.P.R. Arachnida. Vol. 2, Ed. Academiei Republicii Populare
Romne.
11. Fujimoto K., 1990 - Ecological studies on ixodid ticks 7. The effects of humidity on
oviposition development and survival of Ixodes ovatus Neumann (Acarina,
Ixodidae). Jap. J. Sanit. Zool., 41, 331.
12. Hussein S.H., Mustafa B.E., 1987 - Temperature and humidity effects on the life
cycle of Haemaphysalis spinulosa and Rhipicephalus simus (Acari: Ixodidae). J.
Med. Entomol., 24, 77.
13. Ioni Mariana, 2003 - Cercetri privind ecologia familiei Ixodidae n unele zone
subcarpatice; aspecte epidemiologice ale parazitozelor ce pot fi transmise de
acestea. Tez de doctorat, Bucureti.
14. Iwuala M.O.E., Okpala I., 1977 - Egg output in the weights and states of
engorgement of Amblyomma variegatum and Boophilus annulatus. Folia
Parasitol., 24, 162-172.
15. Knlle W., Wharton G.W., 1964 - Equilibrium humidities in arthropods and theier
ecological sifnificance. Acarol., 6, 299.
16. Ouhelli H., Pandey V.S., Choukri M., 1982 - The effects of temperature, humidity,
photoperiod and weight of the engorged female on oviposition of Boophilus
annulatus. Vet. Parasitol., 11, 231-239.
17. Van Der Lingen F.J., Fourie L.J., Kok D.J., Van Zyl J.M., 1999 - Biology of Ixodes
rubicundus ticks under laboratory conditions: observations on oviposition and
egg development. Exp. Appl. Acarol., 23, 513-522.

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IN VITRO ACTIVITY OF A PYRETHRINES


CONDITIONING AGAINST IXODIDAE
AND ARGASIDAE TICKS
R.T. CRISTINA*, I. OPRESCU,
Lidia CHIIMIA, Eugenia DUMITRESCU

In vitro activity of Piretrin extra, a pyrethrum (0, 05g-10.) and


permethrine (0, 20g-10) A.U.V. association, against Ixodidae and Argasidae
ticks study it is proposed.
Parasitic species identified after Babos, Feider and Estrada-Pea
for Ixodidae spp., and after Cosoroaba, for Argasidae spp., were
Dermacentor marginatus, Haemophysalis punctata, respectively Argas
persicus.
Collected ticks were maintained for 48 hours before experiments to
211C at 555 % relative moisture and for 14h: 10h (light/dark)
photoperiod, and four experimental and one control lot of 100 females and
100 males (for each lot) constituted.
Ticks were scattered in Petri plates (simulating living conditions)
and the studied product was uniformly dusted over plates.
Ticks watching accomplished at 12, 24, 36, 48 (for D. marginatus
and H. punctata) respectively 72 hours (for A. persicus) after contact,
followed ticks viability following the locomotors activity, amplitude and
locomotors appendices frequency, ortho and versostasys and death.
Results for treated D. marginatus lots concluded that females are
more resistant than males. 15% mortality ratio for females in comparison
with 16,5% for males at 12h, 23, and 5% comparatively with 22% at 24h
respectively 73%, for females versus 82% for males at 36h. After 48 hours,
registered mortality was of 100% for both genders.
In case of H. punctata lot, results shows again that males are more
sensitive than females, their absolute mortality registering after 36 hours.
In A. persicus lots, death of all individuals supervene in 72 hours,
but is to notice that, 49% of tick population were found dead after 12 hours.
Analysing comparatively it can observed that Argasidae are more resistant
to the studied product than Ixodidae.
In case of D. marginatus and H. punctata eggs, experiments
observed viability of 1000 eggs for each lot. Results showed in all cases,
that after dusting with tested compound, embryonic mortality was absolute,
comparatively with control lots were hatching rate was of 100%.

*
Corresponding author: Prof. Romeo Th. Cristina, PhD
Faculty of Veterinary Medicine Timioara, RO
Veterinary Pharmacology and Pharmacy Dept.
e-mail rtcristina@yahoo.com

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The in vitro tested product proven a very good contact acaricidal


activity against Ixodidae and Argasidae ticks, dead of Dermacentor and
Haemaphysalis individuals supervened in 24 to 48 hours.

Key words: ticks, pyrethrines, in vitro, efficacy

Ticks put control problems mainly because of them vector role of


Anaplasma, Babesia, Cowdria, Theileria and for other nosogenic agents, and for
them direct action upon productions, blood loss and/or them toxic secretions (2, 8,
11).
Fight against ticks is fundamentally an economic issue and success
demands right conception and execution. Therapeutic arsenal against ticks is
wide, from the avoided (because of them disadvantages) old mineral and
organoclorurates (e.g. Toxaphene, Gamatox, Pecusanol) to organophosphorates:
dympylate (Diazinon, Dimpygal), dioxathion (Delvan), coumaphos (Asuntol),
phoxim (Sebacil), quintiofos (Oxinothiophos, Bacdip), chlorfenvinphos (Supona),
carbamates: Propoxur (Sevin, Carbaryl), Bendiocarb, Methomyil. Also the recent
formamidines: chlorphenamide (Clordimephorm, Galeron), amitraz (Taktic),
cymiazole and fermentation compounds (Spinosad), macrolides: Doramectin
Eprinomectin, Ivermectin, Milbemycin-oxime, Moxidectin, Selamectin or
phenylpirazoles (Fipronil) are widely used in the fight against acari.
To these groups is joining also the pyrethroid group from first: (e.g.
Allethrin, Bioallethrin, Permethrin, Phenothrin, Resmethrin and Tetramethrin)
and second: (-Cypermethrin, Cyfluthrin, Cyhalothrin, Cypermethrin,
Deltamethrin, Fenvalerate, Flucythrinate, Flumethrin and Fluvalinate),
generation substances.
Comparatively with the up mentioned ones, this presents, the advantage of
a rapid environmental degradation, low bioaccumulation tendency, and toxicity in
birds and mammals, and not in last line, lack of restrictions for meat and milk
products (3, 6, 7, 8).
*

Present work follow the in vitro activity of an acaricidal conditioning on


Ixodidae and Argasidae ticks, proposing two objectives:
1. In vitro efficacy, against: females, males, and eggs of Ixodidae spp.
2. In vitro activity against Argasidae spp. ticks (sexually undifferentiated).
Experiments were realised in the some time interval and in the same
conditions respecting known methodology of other scientific researches (9, 10).

MATERIALS AND METHODS

Piretrin extra (Phylaxia-Pharma RT) is an A.U.V. association, tested,


approved and recently introduced in our country.

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The studied product is an association of 0,05g pyrethrum (Fig. 1) and 0,20g


permethrine (Fig. 2), and silicone dioxide (0,005g), benzocaine/norcaine (0,001g),
and talcum ad 1000g as excipients, conditioned like dusting powder.

Structure:

O
H3C CH3
H3C CH3 CH2CH CHCH CH2
COO O
H
Cl
H3C CH3
H H C CH COOCH2
C C

R H Cl

Fig. 1 Pyrethrine (C21H28O3) Fig. 2 Permetrine (C21H20Cl2O3)


Acid (2, 2-dimethyl-3-(2-methyl-1-propenyl)- (Sin. Ambush, Ectiban, Pouce)
cyclopropane-carboxylic) 2methyl-4oxo Acid 3-(2, 2-dichloroethenyl)-2, 2-dimentyl-
3(2, 4-pentadienyl) -2-cyclopenten-1-yl clopropane-carboxylic-(3-phenoxyphenyl)
ester methyl ester

The products compounds are efficiently used in many European countries,


because of them low risk and efficient integrated arthropods control in humans
and animals.
Activity of the two components assures long time acaricidal activity and
efficacy, preventing the appearance of resistances. Beside of its quick and time
constant effect, the association has a permanent alarm activity, acting as an
invasion repellent (6, 7).
Piretrin extra is recommended in louse parasitism against: Haematopinus
eurysternus, H. suis, Linognathus oviformis, L. setosus), malofages in birds and
mammals: Menopon gallinae, Goniocotes gallinae, Lipeurus caponis, Trinoton
anserinus, Damalinia bovis, Bovicola ovis, Trichodectes canis, Gyropus ovalis,
Gliricola porcelli, against Argasidae spp.: A. persicus, A. reflexus and Ixodidae
spp.: I. ricinus, Rhipicephalus spp., Dermacentor spp., Haemaphysalis spp.
For the destruction of kitchen bugs: Blatta orientalis, Blattella germanica,
bedbugs: Cimex lectularius, C, hemipterus, and ants: Formica rufa, F. fusca, F.
sanguinaea the product is also recommended.
Piretrin extra is applied in thin coating in places where insects living, and
in them access areas and routes. For 1 m surface is required 5-10g of powder, In
case of external parasitism skin of animal will be dusted, operation being
repeated, if required, to 2-3 weeks. In case of intoxication, atropine is the specific
antidote.

Ticks collecting and identification


Ticks naturally collected, in March-April 2005, from sheep from Timi
County (western part of Romania), respectively from 9 traditional aviaries were
placed in paper bags and brought in laboratory where they were transferred into
plastic jars with sieve cowls.

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Both, Ixodidae and Argasidae ticks were maintained for 48 hours before
assays at 211C to 555%, Relative Moisture, to a 14h: 10h (light/dark)
photoperiod.
Parasitic species identified after Babos (1) and Feider (4) and after
Estrada-Pea (4) were Dermacentor marginatus and Haemophysalis punctata
predominant species in western part of Romania, respectively Argas persicus1,
after Cosoroab (2).

Experiment I

Experimental lots
Four experimental lots and one control (C) were constituted, of 100 females
and 100 males, for each lot and for each parasitic species DM1 and DM2
(Dermacentor marginatus), respectively HP1 and HP2 (Haemaphysalis punctata).
The arthropods were scattered on filter paper and humidified wool
(stimulating body conditions), placed in Petri plates.
After 48 hours, the studied product was dusted, uniformly over all surfaces
of Petri plates (Figures 3 to 6).

Fig. 3 Females of Dermacentor marginatus Fig. 4 Males of Dermacentor marginatus


(dusted Petri plate) (dusted Petri plate)

Fig. 5 Females of Haemaphysalis punctata Fig. 6 Males of Haemaphysalis punctata


(dusted Petri plate) (dusted Petri plate)

1
For testings were used unsexed adults (sex knowing being unnrelevant in this case).

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Readings
Ticks watching accomplished (with the stereoscopic magnifier at 20 x 2,5)
at 12, 24, 36, respectively 48 hours after contact with antiparasitary powder
observed ticks viability following: the locomotors activity; amplitude and
locomotors appendices frequency; ortho and versostasys and death of the
acarians after the conceived key presented in Table 1.
Table 1
Evaluation of tick locomotors activities

Ortho or
Large Large Versostasys Exitus
versostasys
movements movements Very low limbs Motility
Slow limb
hipermotility Slow motility movements absence
movement
++++ +++ ++ + -

Experiment II
Lots
Four experimental and one control (C) of 100 individuals for each lot AP1
and AP2 (Argas persicus) were constituted and placed on humidified filter paper.
After 48 hours Piretrin extra was dusted uniformly over all surface of Petri
plates and argasides viability was expressed in the same parameters like above.
Mortality percent was calculated.

RESULTS AND DISCUSSION

In Tables 2, 3, and 4 is illustrated the mortality evolution in experimental


lots. Analysing the results for treated Dermacentor marginatus lots was
concluded that females are more resistant than males: 15% mortality ratio for
females by comparison with 16,5% for males at 12h, 23,5% by 22% at 24h
respectively 73% for females versus 82% for males at 36h. After 48 hours,
registered mortality was of 100% for both genders.
In case of Haemophysalis punctata lot, results have shown again that males
are more sensitive than females, their absolute mortality installed after 36 hours.
In case of Argas persicus lot, a death of all induals supervenes in 72 hours,
but is to notice that 49% of tick population found dead after 12 hours.
In case of Dermacentor marginatus and Haemaphysalis punctata eggs,
experiments were accomplished observing viability of 1000 eggs for each lot.
Results showed in all cases, that after dusting with tested compound,
embryonic mortality was absolute, comparatively with control lots were hatching
rate was of 100%. However, the action against eggs is very good; this fact has
only scientific information2, the eggs identification in nature being a utopia with
any applicability in the field practice.

2
Even if, in nature will be possible eggs identifications, the product use it is not recommended acaricidal
activity being unselective.

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Table 2
Situation of sexed Dermacentor marginatus lots treated in vitro with Piretrin extra

Treated Dermacentor marginatus Control lot


Males Females
Hours 12h 24h 36h 48h
12h 24h 36h 48h 12h 24h 36h 48h
DM 1. 15 5 63 17 17 8 51 24 - - - -
DM 2. 18 6 57 19 13 9 48 30 - - - -
Average% 16,5 22 82 100 15 23,5 73 100 - - - -
Note: DM: Dermacentor marginatus

Table 3

Situation of sexed Haemophysalis punctata lots treated in vitro with Piretrin extra

Treated Haemophysalis punctata Control lot


Males Females
Hours 12h 24h 36h 48h
12h 24h 36h 48h 12h 24h 36h 48h
HP 1. 72 28 - - 64 26 10 - - - - -
HP 2. 67 33 - - 51 38 11 - - - - -
Average % 69,5 30,5 100 - 57,5 32,0 10,5 100 - - - -
Note: HP: Haemophysalis punctata

Table 4

Situation of Argas persicus lots treated in vitro with Piretrin extra

Treated lots Adults 100 viable individuals/lot Control lot 100 viable individuals/lot
Hours 12h 24h 36h 48h 60h 72h 12h 24h 36h 48h 60h 72h
AP1. 59 30 19 12 10 - - - - - - -
AP2. 39 23 19 10 9 - - - - - - -
Average % 49,0 26,5 19 11 9,5 100 - - - - - -
Note: AP: Argas persicus

CONCLUSIONS

The in vitro tested product proved to be a very good contact acaricidal


against Ixodidae and Argasidae ticks, dead of Dermacentor and Haemaphysalis
individuals supervened in 24 to 48 hours.
The Ixodidae females are more resistant comparatively with males (due
probably because of them size) dying after 36h (for Haemaphysalis females),
respectively 48 hours (for Dermacentor females).
Analysing comparatively it can ascertained that Argasidae are more
resistant to the studied product than the Ixodidae.

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REFERENCES

1. Babos S., 1964 - Die Zeckenfauna Mitteleuropas. Akademiai Kiado, Budapest.


2. Cosoroab I., 2000 - Parazitologie veterinar. Acarioze i entomoze. Ed. Mirton,
Timioara.
3. Davey R.B., 1995 - Efficacy of topically applied pyrethroids against eggs of
Boophilus microplus. J. Agric. Entomol. 12, 1, 67-73.
4. Estrada-Pea A., Bouattour A., Camicas J.L., Walker A.R., 2004 - Ticks of
domestic animals in the mediterranean region. A guide to identification of
species. ICTTD.
5. Feider Z., 1965 - Fauna R.P.R. Arachnida. Vol., 2, Ed. Acad. RPR.
6. Sangster N.C., 1996 - Pharmacology of anthelmintic resistance. Parasitol. 113, 201-
216.
7. Samish M., 2000 - Biocontrol of ticks. Ann. N. Y. Acad. Sci. 916, 172-178.
8. Samish M., Glazer I., 1991 - Killing ticks with parasitic nematodes of insects. J.
Invert. Pathol. 58, 281-282.
9. Stendel W., Fuchs R., 1982 - Laboratory evaluation of Flumethrin, a new synthetic
pyrethroid for the control of one- and multi-host ticks. Vet. Med. Rev. 2, 115-129.
10. *** - www.unine.ch/tiques
11. *** - Parasitology Laboratory. Acari (ticks and mites) in Biology 625. Animal
parasitology, [on-line], http://www.ksu.edu/parasitology/classes/625arth47.html.

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REZULTATE AL INVESTIGAIILOR ARHEOFAUNALE


NTR-O CLDIRE DIN ORAULUI ROMAN NAPOCA
I O AEZARE RURAL DIN LOCALITATEA
UCEAGU (JUD. CLUJ)
RESULTS OF ARCHAEOFAUNAL INVESTIGATIONS IN A ROMAN CIVILIAN
BUILDING IN THE ROMAN TOWN NAPOCA AND A RURAL SETTLEMENT
IN SUCEAGU VILLAGE (CLUJ COUNTY)

A. GUDEA, F. STAN F.,


A. SOCACIU, A. DAMIAN
U.S.A.M.V. Cluj-Napoca

The present study presents briefly the results of two


archeozoological investigations (as part of a much more complex study
concerning the roman fauna in the province of Dacia) carried on two small
archaeofaunal samples from a civilian building in the roman city of Napoca
and another roman rural settlement from Suceagu village, 10 km far from
Napoca.
These two archaeological sites provided us small samples of animal
bones (below 100 pieces) and the results showed in case of the civilian
building in Napoca that the only species identified is cattle(Bos taurus). A
MNI of 2 was established and, based on the osteometrical data, a hight of
111 and 124 cm recalculated (based on the Matolcsis formula). Both
identified individuals were over 2,5 years old. The sample from the rural
settlement from Suceagu is larger, showing the existence of cattle,
capriovids, horse and dog (as domestic species) and roe deer (as game).
Statistically, the predominance of cattle is shown as primary meat-source
species, followed by capriovids and horse. Osteometrical data is available
in case of cattle, showing the existence of an male individual with 131 cm
high (Matolcsi) and in case of one Ovis individual of 65 cm high.

DATE PRELIMINARE

Studiile arheofaunale sau arheozoologice sunt importante att din punct de


vedere al datelor de ordin morfologic care se pot desprinde prin analizele
resturilor de oase descoperite n spturile arheologice efectuate n diferitele
obiective ct i prin datele de ordin socio-economic desprinse ca element de
ilustrare al nivelului vieii economice prin prisma tiinei creterii animalelor.
Datele analizelor sunt cuprinse ntr-un studiu amplu al primului autor,
studiu ce vizeaz diferitele aspecte ale creterii i ngrijirii animalelor n epoca
roman.
Materialul osos recoltat provine dintr-o prim sptur efectuat ntre anii
1995-1996 de ctre dr. Coci Sorin (Institutul de Arheologie Cluj Napoca) ca
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urmare a demolrii fostei sli a clubului de box din centrul oraului Cluj Napoca.
Obiectivul studiat se gsete i astzi n apropierea parcului I.L Caragiale, pe
strada Victor Deleu, vis-a-vis de Liceul de Muzic i constituie una din atraciile
turistice amenajate din centrul oraului. Primele spturi (de unde ne-a parvenit
materialul osos- singurul recoltat n decursul acestor investigaii) au scos la iveal
pri ale unor cldiri datate n ca fiind de secol XI-XII iar ntr-un strat mai adnc
cteva pri ale unor cldiri atribuite perioadei romane, cldiri a cror construcie
era predominant fcut din lemn cu ziduri mici de piatr. Eantionul osos este
foarte mic ca numr de oase, fazei romane fiindu-i corespondente doar 36 de
fragmente iar celei de secol XI-XII-130.
Aezarea rural roman de lng localitatea clujan Suceag a fost
investigat printr-o sptur de control efectuat n cursul anilor 1995-1997 tot
sub coordonarea dr. Coci Sorin. Eantionul osos recoltat cuprinde 612 oase, din
care doar 50 oase identificabile provin din stratul datat pentru epoca roman,
restul de 443 sunt provenite din stratul de secol IV p.Ch. Rezultatele investigaiei
osteologice vizeaz aici doar acest mic eantion atribuit perioadei romane.

MATERIAL I METOD

Materialul osos igienizat la locul de recoltare a fost preluat de la locul de


depozitare i transportat la Laboratorul de Anatomie Comparativ a Facultii de
Medicin Veterinar Cluj-Napoca.
Analiza arheofaunal a constat n separarea oaselor pe zona anatomice i
ndeprtarea fragmentelor neidentificabile, urmat de identificarea speciilor prin
observaii anatomice. S-a fcut repartizarea la membrul aparintor (n scopul
determinrii numrului minim de indivizi. S-au fcut observaii cu privire la urmele
de prelucrare uman. Aprecierile cu privire la vrst s-au fcut prin observarea
stadiului de epifizare a oaselor ct i asupra stadiului de erupie i uzur dentar.
Msurtorile s-au fcut utiliznd msurtorile etalon folosite n osteometrie.

REZULTATE I DISCUII

Eantionul de dimensiuni foarte mici recoltat din cldirea din str. Deleu-
Napoca 36 fragmente - nu este foarte reprezentativ, mai ales datorit faptului c
au fost identificate oase ale unei singure specii. Pentru fragmentele identificate
atribuite bovinelor- se pot trage cteva concluzii a cror interpretare poate fi
fcut doar ntr-un context mai larg - cel al comparaiilor datelor metrice cu cele
surprinse n cazul altor situri contemporane.
Este destul de dificil de apreciat numrul minim de indivizi din acest
context. Singurele date ce se pot obine sunt cele referitoare la vrst - individul
sau indivizii identificai sunt de peste 2,5 ani. O oarecare departajare ar fi posibil
datorit existenei a 2 fragmente indiciu al unei vrste de peste 3,5 ani i alte 2
indiciu al unei vrste de peste 2,5 ani, aadar fiind posibil o apreciere a unui

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minim de 2 indivizi. Datele metrice surprind de asemenea 2 indivizi, din care unul
a fost apreciat drept femel, cu o talie de 1240 mm iar altul de 1109 mm
(Boessnek).
Pentru eantionul provenit din aezarea rural de la Suceagu a crui
dimensiune mai mare a permis identificarea mai multor specii ct i desprinderea
a mai multor date, rezultatele sunt concentrate iniial sub form tabelar pentru ca
mai apoi s fie trase cteva concluzuii referitoare la acesta.

Specii identificate
Specii domestice
Bos Taurus ++
Ovis aries ++
Capra hircus +
Equs cabalus ++
Sus scrofa domesticus ++
Canis familiaris ++
Specii slbatice
Capreollus capreollus ++
++ specie identificat cert
+ specie a crei prezen este posibil

Specia NISP % MNI


Rumegtoare mari 32 64 3
Ovicaprine 8 16 3
Sus scrofa dom 2 4
Equs caballus 6 12
Canis familiaris 2 4
Total 50

Bovine (Bos taurus)


Numrul foarte mic al fragmentelor osoase atribuit acestei specii nu
permite concluzii tranante. Putem aprecia doar posibila provenien a oaselor
identificate de la minim 2 sau 3 indivizi. Datele despre stadiul de osificare
surprinse la nivelul oaselor lungi arat 2 indivizi de peste 2-2,5 ani din care unul
de peste 3,5-4 ani i probabil un alt individ sub 2,5 ani. Datele metrice parial
surprinse n cazul metatarsului permit aprecierea unei talii de 1315 mm
(Matolcsi), cel mai probabil provenit de la un mascul. Urmele de prelucrare indic
destul de clar utilizarea speciei n scop alimentar.
Ovicaprine (Ovis et Capra)
Concluziile ce se pot surprinde pe baza materialului analizat sunt foarte
puine. Vorbim despre existena a minim 2 sau 3 indivizi ai grupului ovicaprinelor
(fr ca specia s poat fi desemnat), dintre care 1 sau 2 sunt de peste 2,5-3 ani i
un alt individ este sub 18-21 luni. Unul din indivizii de peste 2,5-3 ani are o talie
de 65 cm.

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Cabaline
Nu se poate aprecia dect existena unor indivizi robuti (apreciere
subiectiv).

CONCLUZII

n ciuda faptului c n lucrarea de fa avem de-a face cu dou eantioane


mici ca dimensiuni, acestea ne permit cteva concluzii de ordin general, att n
privina datelor de ordin morfologic desprinse din studiul asupra oaselor
descoperite ct i n privina elementelor socio-economice ce deriv din acestea.
Concluziile desprinse sunt importante att ca date de sine stttoare (date
biometrice, osteometrice) ,atunci cnd facem referire la rezultatele pentru fiecare
sit arheologic n parte, ct i ca date integrate, alturate celor desprinse din
analizele arheozoologice efectuate n situri contemporane.
Se poate spune c n ambele situri specia predominant este cea bovin
(rumegtoarele mari) a cror exploatare se fcea n scop alimentar (carne) dar i
utilitar (munc, produse). Urmtoarele specii ca importan sunt reprezentate de
ovicaprine i suine. Speciile secundare (cabaline, canide) sunt identificate n
eantionul de la Suceagu i aspectele surprinse n privina lor nu sunt neobinuite
pentru epoca roman. Speciile slbatice- cpriorul (n cazul eantionului de la
Suceagu) sunt slab reprezentate numeric, indiciu al unei lipse de preocupare n
privina vnatului, ns dovad a existenei acestei specii n biotopurile
nconjurtoare.
Elementele de morfologie care se pot desprinde arat n cazul bovinelor
valori ale taliilor ce ncep de la 110 cm pn la 130 cm, elemente care arat, la fel
ca i n cazul altor situri de epoc roman, creterea valorii taliilor bovinelor fa
de epoca precedent.
n privina ovicaprinelor s-a putut recalcula o singur talie pentru un individ
matur- 65 cm- valoare care este nscrs n tendina cresctoare a valorii taliilor
pentru ovicaprinele epocii romane, rod al ameliorrii speciei.
Pentru suinele domestice i pentru cabaline datele surprinse n cazul celor
dou situri nu sunt relevate.

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17. Udrescu, M.S., L. Bejenaru & C. Hriscu, 1999 - Introducere n arheozoologie. Iasi:
Corson.
18. Von Driesch, A., 1976 - Das Vermessen von Tierknochen aus Vor- und
Fruhgeschichtlichen Siedlungen. Mnchen.

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CALITATEA HRANEI I IMPACTUL ACESTEIA


ASUPRA RELAIEI LUNGIME-GREUTATE
LA JUVENILII RACULUI DE RU
FOOD QUALITY IMPACT UPON THE RELATION BETWEEN LENGTH
AND WEIGHT OF YOUNG NOBLE CRAYFISH

L. MIRON
U.S.A.M.V. Iai
Manuela MIRON
Univ. AL.I.CUZA Iai

Three varieties of food were tested during this study: commercial


trout pellets, fresh fish ,carrots and potatoes (ready-cooked vegetables).
Food quality impact upon the relation between length and weight was
studiet of young noble crayfish Astacus astacus.The alometry rate of the
relation between length and weight of the differents clusters is
approximately 3.The ratios of relative condition are greater than 1.
However, the maxima value is observed for crayfish feed with commercial
trout pellets.During this test, water analysis revealed some traces of
nitrogenous elements in the troughts where crayfish were feed with trout
pellets.

Key words: crayfish, relation length-weight, food quality

n aceast lucrare, a fost realizat un studiu experimental al impactului


hranei asupra creterii juvenililor de rac de ru i asupra calitii apei, a diferitelor
tipuri de alimente disponibile pe pia: legume prefierte (morcov i cartofi) ca
surs de proteine vegetale, pete proaspt -ca surs de proteine animale i hran
compactat sub form de granule, ca aliment complet (2, 4, 7). Rezultatele
experimentelor au vizat un dublu obiectiv:
- testarea unor alimente susceptibile de promovare a astaciculturii;
- determinarea performanelor de cretere la puii de rac de ru n mediul
artificial.

MATERIAL I METOD

Racii folosii pe durata experimentului, la Staiunea Biologic Potoci, au fost


capturai din lacul Bicaz. Lungimea total a acestor exemplare (msurat de la
rostru la telson) a variat ntre 50 i 60 mm. Instalaia de cretere a juvenililor de
rac a fost compus din cuve din fibr de sticl (2m lungime;0,5 m lime; 0,5 m
adncime) alimentate cu ap de izvor (4 litri/min). Aceste cuve au fost prevzute
cu adposturi etajate din P.V.C., cu diametrul de 40 mm fabricate local,

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respectndu-se biologia acestei specii, care n tinpul zilei st retras n refugii.


Masa pentru distribuirea alimentelor a fost aezat ntre adposturi.
Racii au fost repartizai n 6 loturi a cte 20 de exemplare n 6 cuve. Raia
de alimente a fost stabilit astfel:
- 100 g pete proaspt pentru loturile A1i A2;
- 100 g legume prefierte pentru loturile A3 i A4;
- 100 g granule pentru pstrv pentru loturile A5 i A6.
Lungimea i greutate racilor din fiecare lot, a fost controlat la fiecare 15
zile. Temperatura, pH-ul i coninutul n oxigen dizolvat au fost msurate in-situ.
Ceilali parametri urmrii au fost determinai n laboratorul de analiz a calitii
apei.
Pentru a pune n eviden impactul calitii alimentelor asupra creterii
racilor, parametrii studiai s-au referit la relaia dintre lungime i greutate a
indivizilor din diferitele loturi. Aceast relaie este:

P= a. x Lb (1)

unde P reprezint greutatea individului, a=constant iar b este coeficientul


de cretere relativ sau coeficientul de alometrie. Plecnd de la aceast formul,
substituirea valorilor lungimii = Lcu lungimile medii observate n cursul
experimentului, permite obinerea valorii greutii medii calculate. Coeficientul de
condiie relativ este raportul ntre greutatea observat i greutatea calculat.
Acest coeficient se traduce prin condiia fizic a animalului i prin starea de
bonitate.

REZULTATE I DISCUII

Relaia lungime- greutate a celor 6 loturi de raci a relevat o variabilitate a


creterii n funcie de alimentul administrat. Pe timpul derulrii experimentului s-a
remarcat existena a trei faze de cretere:
- A - debuteaz cu nceputul experimentului i dureaz 2 sptmni;
este faza de adaptare a racilor la mediu;
- B - faza se ntinde pe parcursul a 4 sptmni i coincide cu perioada
de cretere a racilor (nprlire); faza are un vrf de cretere care
variaz ntre 75% i 90% pentru cele 6 loturi de juvenili;
- C - faza dureaz 16 sptmni i se caracterizeaz prin diminuarea
indicelui de cretere ce coincide cu debutul fazei de oprire a creterii
racilor, faz numit internprlire
Corelaia ntre greutate i lungime la raci, ne permite s apreciem gradul de
adaptare la mediu. Ca i la peti, aceast relaie este foarte important, de tip
cretere (1). Trebuie remarcat c pentru creterea n lungime, a fost pus n
eviden o variaie a vrfului de cretere n funcie de loturi. Astfel la diferitele
loturi, cuplul lungime- greutate a descris o curb de tip cretere, n tabelul 1
fiind prezentai parametrii acestei relaii.

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Tabelul 1
Parametrii relaiei lungime- greutate la diferite loturi de raci n cretere

Loturi Model Corelare


A1 P= 3x 10-5 L3,0052 R = 0,97
A2 P= 3x 10-4L2,4261 R = 0,89
A3 P= 2x 10-5L3,1295 R = 0,97
A4 P= 4x 10-5L2,9520 R = 0,89
A5 P= 2x 10-5L3,1680 R = 0,98
A6 P= 3x 10-5L3,1302 R = 0,88

Aceste rezultate arat c pentru diferite loturi, valoarea alometriei este


foarte aproape de 3. Creterea este deci izometric. Totui trebuie notat c
valoarea alometriei la loturile alimentate cu granule de pstrv, este uor
superioar celorlalte loturi iar indivizii acestor loturi se afl ntr-o foarte bun
stare de sntate (3,6).
Creterea ponderal i coeficientul de condiie a diferitelor loturi este
prezentat n tabelul 2.
Tabelul 2
Creterea ponderal a diferitelor loturi de raci

Coeficientul de condiie
Loturi Cretere ponderal (g)
relativ
A1 9,7 1,01
A2 9,9 1,01
A3 11,7 1,07
A4 10,9 1,05
A5 13,2 1,25
A6 14,5 1,30

Compararea valorilor prin testul T Student nu a artat diferene


semnificative ntre loturile cu acelai regim alimentar (o probabilitate mai mic
de 5%). Ctigul n greutate la exemplarele hrnite cu granule de pstrv este
semnificativ superior loturilor hrnite cu legume prefierte i cu pete proaspt.
Creterea n lungime a nregistrat rezultate mai sczute la racii crescui cu pete
proaspt. Loturile de raci hrnii cu legume prefierte prezint valori intermediare.
Datele confirm superioritatea granulelor de pstrv ca aliment complet ce
asigur o cretere optim care reflect i o stare bun i o condiie fizic bun a
animalelor. Examinnd compoziia granulelor de pstrv, constatm c ele conin
produi azotai de origine vegetal, co- produi de transformare a cerealelor,
produi i co- produi de transformare a zaharurilor, ulei, grsimi, substane
minerale i vitamine. Aadar hrana granulat reprezint un aliment ce asigur
nevoile energetice ale racilor. Va fi interesant de pus la punct un aliment
comercial pentru raci, care s permit obinera unor performane de cretere.

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Procentul de mortalitate a fost urmrit pe parcursul experimentului mai ales


n faza de adaptare a racilor, care au fost introdui ntr-un mediu nou diferit de cel
de origine, aceast modificare putnd induce moartea la crustacee, mai ales n
acest stadiu al dezvoltrii.
Regimul pe baz de granule de pstrv asigur un procent de 95% de
supravieuire. Hrnirea cu pete permite supravieuirea ntr-un procent de 85% iar
hrnirea cu legume conduce la un procent de 70% supravieuire a subiecilor.
Impactul calitii apei
Controlul calitii fizice i chimice a apei s-a fcut la interval de 15 zile.
Valorile parametrilor fizico- chimici au rmas constante n toate cuvele de
cretere. Valorile medii ale temperaturii, pH-ului i oxigenului dizolvat au fost:
13,6oC; 7,2; respectiv 8,4 mg/l. Analiza apei a confirmat starea satisfctoare
de calitate n toate cuvele experimentale, ca i absena elementelor azotate. n
tabelul 3 sunt prezentate valorile parametrilor msurai.
Tabelul 3
Valorile medii ale parametrilor fizico- chimici n apa de izvor

Factori Ap
Cuva1 Cuva2 Cuva3 Cuva4 Cuva5 Cuva6
analizai izvor
Loturi A1 A2 A3 A4 A5 A6
ToC 13,6 13,6 13,6 13,6 13,6 13,6 13,6
PH 7,3 7,3 7,3 7,3 7,3 7,3 7,3
O2 mg/l 8,5 8,3 8,3 8,3 8,3 8,3 8,3
OxMnO4K
0,35 0,45 0,5 0,5 0,5 0,6 0,65
Mg/lO2
Ca mg/l 70 70 70 70 70 74 74
Mg mg/l 32 32 32 32 32 33 33
Sulfat.mg/l 34,5 34,5 34,5 34,5 34,5 34,5 34,5
Clor mg/l 14,0 14,3 14,3 14,2 14,2 14,2 14,2
NO3 mg/l 0 0 0 0 0 urme urme
NO2 mg/l 0 0 0 0 0 urme urme
NH4 mg/l 0 0 0 0 0 urme urme

Apa este caracterizat printr-o temperatur i pH. constante. Valorile


coninutului n oxigen dizolvat i n calciu sunt optime pentru dezvoltarea racilor
Astacus astacus (1, 2, 6) . S-au nregistrat doar slabe urme de derivai azotai n
cuvele unde hrnirea loturilor de juvenili s-a fcut cu granule de pstrv.

CONCLUZII

Experimentul a avut ca scop dezvoltarea creterii intensive a racilor de ru


Astacus astacus L.i definirea unor metode de gestionare ct mai rentabile.
S-a demonstrat superioritatea hranei granulate folosite i n creterea
pstrvului, fa de petele proaspt i legumele prefierte, determinnd o mai bun
cretere n lungime i greutate. Hrana granulat prin compoziia sa complet,
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favorizeaz creterea racilor i starea fizic a exemplarelor, fapt dovedit i de


procentul de supravieuire. Remarcm necesitatea fabricrii unui aliment specific
pe baza produselor locale care s rspund nevoilor energetice ale racilor pe
parcursul ciclurilor de dezvoltare.
n ceea ce privete calitatea apei, nici unul din alimentele testate nu prezint
un efect poluant. Prezena urmelor de nitrii, nitrai i amoniac n cuvele n care
s-a folosit hrana granulat, trebuie s incite la noi cercetri pentru verificarea
impactului la o scar mai mare, a acestui tip de hran, asupra calitii apei

BIBLIOGRAFIE

1. Austin, C.M., 1995 - Length-weight relationships of cultured species of Australian


freshwater crayfish of the genus Cherax. Freshwater crayfish 10: 410-418.
2. Gydemo, R., 1988 - Growth of the noble crayfish, Astacus astacus, in a pond-stoking
experiment. Freshwater crayfish 7: 251-258.
3. Henttonen, P., 1993 - Moulting, growth, survival and colour of Astacus astacus L.
juveniles fed diet with and without green plant material and maintained in
individual cages and communal tanks. Freshwater crayfish 9: 426-441.
4. Laurent, P.J., 1989 - Les ecrevisses dans le monde. LAstaciculteur de France nr.
21: 9-17.
5. McMahon, B.R:, 1986 - The adaptable crayfish: mechanisms of hpysiological
adaptation. Freshwater crayfish 6: 65-72.
6. Miron Manuela, 2006 - Compendiu de astacicultur. Ed. Stef. Institutul European,
254p.
7. Pursiainen, M., & al, 1988 - Moulting and growth of the noble crayfish Astacus
astacus in an oligotrophic lake. Freshwater crayfish 7: 155-164.
8. Sevilla, C., 1993 - Biochemical study of the growth of Astacus astacus fed with plant
diets by quantitative variations of muscular DNA and proteins. Freshwater
crayfish 9: 226-234.

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ANALYSIS OF FOOD UTILISED BY THE CRAYFISH


ASTACUS ASTACUS L. FROM BICAZ LAKE
ANALIZA HRANEI UTILIZATA DE RACUL DE RAU ASTACUS ASTACUS L.
DIN LACUL BICAZ

Manuela MIRON
Univ. "AL. I. CUZA" Iai
L. MIRON
U.S.A.M.V. Iai

The food consumed by crayfish (Astacus astacus L.) was investigated


in Bicaz Lake, NE of Roumania, during the ice-free period. The stomach
content of crayfish of different sizes and from different habitats were
examined. Despite a transition from oligotrophic to mesotrophic conditions
in the shallow areas of the lake, the food composition had not changed
markedly since the last study in 1980 (Miron et al., 1983). The majority of
individuals from all size groups contained detritus and/or benthic
macrophytes on all sampling dates. Benthic algae also constituted a
considerable part of their diet, while benthic crustaces, insects, zooplankton
and fish only made up a minor part of the diet. The frequency of crayfish with
empty stomachs was higher during late autumn and during molting than at
other times.

The ecological role of crayfish in benthic communities and their overall


trophic positlon in the lake eco-system are often unknown and overlooked (Mason
1963, Vannote 1963, Momot et al. 1978). However, food choice by Astacus
astacus has been of little interest despite the need for such information when
attempting to evaluate the importance of crayfish in the benthic communities.
We studied food choice and determined the importance of various food
items in the diet of A. astacus in Lake Bicaz in 2004-2005. This lake provides
aproximately, 2 kg/ha (Manuela Miron, 1999) and is probably a good habitat for
A. astacus in Romania today. During the last few decades, there has been a
transition from oligotrophic to mesotrophic conditions in the lake. The feeding of
crayfish was examined in Bicaz lake 10 years ago (Manuela Miron, 2004)
providing comparative data, to reveal long-term changes in feeding. Further, we
discuss the importance of crayfish with respect to the turnover rate of organic
deposits from the littoral habitats.

MATERIALS AND METHODS

Crayfish were sampled during July 2004- June 2005 at Biological Station
Potoci, located near Bicaz lake. In June, July, August and September,

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crayfish were handly collected within a defined boundary. In the shallow (< 1.5 m),
littoral area where the bottom substratum consisted of stones and gravel. Areas
with thin cover of waterweed were avoided. In October, crayfish aere located in
deeper water and sampling was therefore conducted by diving (1,5-10 m).
After sampling, a 10 % formaline solution was injected into the stomach of
each crayfish which was stored in a deep-freezer. A total or 171 individuals (size
4.5-10.0 cm) had their stomach contents (not intestines) examined under a
binocular microscope and the food remains were identified when possible.
Stomach fullness was subjectively evaluated using a numerical index; 0-
designated empty and 4 designated fully extended. The size, sex and carapax
status (stage of molting) were determined for each individual.

RESULTS

Food Items
Throughout the summer, benthic macrophytes and detritus were the main
food components, with macrophytes dominating during early summer and detritus
dominating during late summer. Although that two major food groups were often
found together in the same stomach, extended stomachs (degree 4) often consisted
exclusively of either macrophytes or detritus. Animal remains occurred most
often together with plants, whlle benthic algae always co-occurred wlth
sediments. Plant remains were most often composed of brownish, decaying stems.
Fresh green plants were only occasionally recorded; cccasionally decaying
plants and detritus were hard to differentiate, usually, however, and especially
in extended stomachs, the detritus component consisted of soft mud containing
few larger particles. A large number of animals was found, neither of which made
significant contributions to that diat (Tab. 1). Most frequent were the remains of
the lacustris crustaceans, the larvae and pupae of trichopteran and eurytopic
chironomidae like Chironomus plumosus.
Fish remains and fish scales were frequently recorded in all seasons. Scales
of bleak Alburnus alburnus L. were identified mostly in the medium size. This
fish species was commonly used as bait in the crayfish traps.

Relative importance of food items


Although a large number of pray types were identified from crayfish
stomachs, most of these were only a small fraction of them total volume (Tab. 1).
In September, animal remains were found in more than 60 % of the stomachs,
accounting for less than 30% of volume. In July, the corresponding values were
28% and 7%, respectively. In total, plant remains and detritus were by far the
most important food items, each of these food types accounting for more than 40 %
of the food volume. This change from mainly macrophytes in early summer to
mainly detritus in late summer and autumn is noteworthy.

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Table 1.
Relative amounts (volume %) of various types of prey in stomachs
(volume %) of A. astacus

Food groups June July August September October Mean


Benthic macrophytes 74 59 10 26 20 37.8
Benthic algae - 3 8 9 - 4
Sediment 21 23 71 37 60 42.4
Leaves 1 2 - - - 0.6
Seeds 0.5 2 - - - 0.6
Fish - 0.5 3 9 1 2.7
Trichoptera - - - 5 4 1.8
Chironomidae - 2 2 1 - 1.0
Cyclops 1 - - 1 - 0.4
Bosmina - 0.5 - 0.5 - 0.2
Chitinous fragments 2 2 1 3 3 2,2
Other subjects - 4 2 0,5 1 1,5

Feeding Activity
Stomach fullnes used as a measure of feeding activity, showed considerable
variatlon during the summer. Feeding was most intense around mid-summer and
during early autumn; it was lower in early summer and in October when almost
90% of the sampled crayfish had empty or almost empty stomachs. Although all
individuals caught between June-September came from shallow areas (< 1.5 m
depth), only a single individual was caught at this depth in October. The rest were
collected by a Scuba diver (1.5-10 m). This single individual found in the
shallows, was the only crayfish collected in October that had its stomach full of
food.
No correlations were found between sex or size and stomach fullness.
There was, however, a slight tendency for larger animals to feed more than
smaller animals during June, August and September. In July, the individuals with
empty or almost empty stomachs were significantly larger than then the well-fed
individuals (P < 0.05, t-test). This relationship coincided wlth molting, which
occurred during this time among the largest individuals. All individuals at different
stages of the molting cycle had almost empty stomachs. Feeding activity in egg-
carrying females was not significantly different from the rest of the crayfish
population.
Organic detritus dominated: it was found in 74% of the stomachs; 53%
contained macrophytes, 46% algae, 9% seeds, 15% Chironomidae, and 7%
Nematoda. Fish, zooplankton and other animals were only found in sma11
numbers. The frequency of crayfish with empty stomachs was only 7%. However,
most of these were found in early August.

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DISCUSSIONS

The present results are basically in agreement with other studies concerning
crayfish feeding. They all concluded that benthic macrophytes and detritus are the
major food components (Abrahamsson 1966, Prins 1968, Momot et al. 1978).
Compared with these studies, however, the detritus component in our study
was relatively large (21-71%, mean 42%), while the vascular plant (10-39%,
mean 30%) and benthic algae components (0-9%, mean 4%) were relatively less.
The proportion of animal material was typically ca. 14% (range 3.5-27.5%).
During June and July, benthic macrophytes constituted more than 66%
(volume) of the ingested food, while detritus only accounted for 22%. This
situation was reversed in August-October. Thus during spring, crayfish appear
to feed on decaying plant material left over from previous years, gradually shifting
to detritus which becomes enriched with organic material and bacteria as the
sesson progresses. Furthermore, in late summer, the detritus component contained
large amounts of diatom fragments, providing an additional food source. The
largest proportion of detritus in the stomach semples coincided with periods of
lowered feeding activity (August-September).
The crayfish is a potential fish predator, although fish have been rarely
recorded as food. The fish remains and large fish-scales found in stomach samples
during autumn probably originated as bait in traps used between August-
September. Thus if fish are excluded as a potential food source in nature, then
animal material makes up for only 11% of the total food volume. Fish roe,
crayfish roe and young crayfish were never recorded in stomachs, despite the
crayfish's reputation as a roe-predetor and cannibal.
The nutritive value of the various food types is uncertain. The nutritive
value of detritus probably varies between seasons and between different localities.
Whether or not crayfish benefit from small amounts of animal proteins has also
been debated. Although information on crustaceans is scarce, there is some
evidence that omnivores on a mixed diet grow better than those limited
exclusively to either plant or animal material (Kinne 1959, Covich 1977). With the
exception of detritus and benthic algae no food groups were found to be
significantly linked in this study. Many kinds of food were often found together in
the same stomach, indicating a non-selective feeding mode. There was, however,
a tendency for larger individuals to exploit a single food source (either
macrophytes or detritus), and the increased amount of animal material in smaller
individuals agrees with the findings of Lund (1944) and Vannote (1963).
Large numbers of crayfish only containing detritus were found supporting
the hypothesis that detritus alone may be a sufficient food source for parts of the
sampling period. The value of detritus as food is probably high since it may
contain large amounts of protein (bacteria, decaying plant and animal material)
and minerals.

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Several individuals had fed exclusively on macrophytes, inferring the


presence of cellulase in the crayfish (cf. Lund 1944, Van Weel and Scheer 1970).
This inference is supported by the findings of Moshiri and Goldman (1969), who
found an assimilation efficiency of nearly 50% for crayfish fed solely on
plants.The crayfish is obviously of great importance in the benthic ecosystem, due
to its high densities and omni- and detrivorous feeding habits (cf. Momot et al.
1978). Thus it is probably the main contributor to the benthic fauna.

LITERATURE CITED

1. Abrahamsson, S.A.A., 1966 - Dynamics of an isolated population of the crayfish,


Astacus astacus Linne. - Oikos 17: 96-107.
2. Covich, A.P., 1977 - How do creyfish respond to plants and mollusca as alternate
food sourcea? - Freshw. Crayfish 3: 165-179.
3. Kinne, O., 1959 - Ecological dats on the amphipod Gammarus duebeni. A
monograph. - Veroff. Inst. Meeres Forsh. Bremerh. 6: 177-202.
4. Kjellberg, G. Bunndyr I. Tyrifjordan - In: D. Berge (ed.), Tyrifjorden.156 pp. (In
Norvegian).
5. Lund, H.M.K., 1944 - A study of the food of the crayfish. - Nytt Mag. For Nat. vitensk.
B4: 219-250.
6. Mason, J.C., 1963 - Life history and production of the crayfish, Pacifastacus
leniusculus. M.S. Thetis, Oregon State Univ., Cornvallis. 204 pp.
7. Miron I. & col., 1983 Lacul de acumulare Izvoru Muntelui Bicaz . Monografie
limnologica. Ed. Acad RSR.
8. Miron M., 1999 Cercetari asupra biologiei crustaceelor din lacul Bicaz si a
posibilitatilor de crestere dirijata a unor specii cu valoare economica ridicata.
Teza de doctorat.
9. Momot, W.T., H, Gowing, P.D. Jones, 1978 - The dynamics of crayfish and their
role in ecosystems. - Am. Mid. Nat. 99: 10-35.
10. Moshiri, G.A., C.R. Goldman, 1969 - Estimation of assimilation efficiency in the
creyfish Pacifastacus leniusculus (Dana) (Crustacea: Decepode). - Arch.
Hydrobiol. 66: 298-306.
11. Skurdal, J., T. Qvenild, 1986 - Growth, maturity and fecundity of Astacus astacus in
Norvey. - Freshwater Crayfish 6. Vannote, R.L. 1963. Community productivity
and energy flow in an enriched warm water stream. - Ph.D. Thesis, Michigan
State Univ., E. Lansing.156 pp.
12. Van Weel, P.B., B.T. Scheer, 1970 - Chemical Zoology 5. - In: M. Elorkin
Arthropoda. Acad. Press, New York.

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AGRESIUNE PARAZITAR MULTIPL ASUPRA


ESUTULUI HEPATIC LA CAPRA NEAGR
(RUPICAPRA RUPICAPRA)
MULTIPLE PARASITICAL AGGRESSION ON THE HEPATIC TISSUE
OF THE BLACK GOAT (RUPICAPRA RUPICAPRA)

Otilia COOFAN, Olimpia IACOB,


I. POP, Gabriela RILEANU
U.S.A.M.V. Iai

The research has been made on two cases, the Black Goat species
(Rupicapra rupicapra) who died because of transporting stress.
The hunts support state was showing a deficit, with pronounced
weakening. The general necropsical examination was followed by the
morphopatological examination of the liver, from which samples of tissue
were fixed on formaldehyde 10% and after that were included in paraffin
The samples were sectioned at 5 m, colored through the HEA,
MGG, PAS methods, examined and microphotographed at MC5 with 10,
ob. 10, 20, 40, and Imm. 63 oc.
The histopathological examination revealed recent aggressions on
the hepatic tissue characterized by hemorrhagic, necrotic paths and acute
angiocolitis also hemorrhagic-necrotic paths, a previous aggressions
expressed by chronic angiocolitis and hepatitis.
These modifications are characteristic for the traumatic hepatitis
produced by Cysticercus tenuicollis (cysticercosa hepatitis) during the
hepatical migration, extremely aggressive, metacestod identified along with
other parasitical structures disposed in the hepatical tissue.

Key words: Hunt, Black goat (Rupicapra rupicapra), liver, Cysticercus tenuicollis,
multiple parasitical aggression

Parazitozele hepatice ale vnatului rumegtor, (capra neagr - Rupicapra


rupicapra), afeciuni cu evoluie cronic, insidioas, rareori acut, determin
perturbarea proceselor metabolice, influennd negativ procesul de cretere la
tineret, scderea rezistenei i reactivitii la boli, diminuarea produciilor la
animalele adulte i frecvent, mortalitate.
Condiiile de via ale vnatului sunt din ce n ce mai dificile ca urmare a
limitrii spaiului vital i n consecin aglomerrile teritoriale contribuie la
apariia i perenizarea parazitozelor (3).
Carnivorele slbatice, gazde definitive pentru numeroase specii de cestode,
sunt responsabile pentru poluarea parazitar a suprafeelor punabile i apariia
metacestodozelor hepatice la vnatul rumegtor. Agresiunea hepatic, exercitat

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de mai multe specii de paraziti, se desfoar de obicei, simultan, genernd o


aciune concertat cu efect letal asupra vnatul rumegtor.
Scopul cercetrilor este relevarea agresiunii parazitare multiple asupra
esutului hepatic la capra neagr (Rupicapra rupicapra) i consecinele asupra
vnatului rumegtor.

MATERIAL I METOD

Cercetrile au fost ntreprinse asupra a dou exemplare de capra neagr


(Rupicapra rupicapra) sucombate n urma stresului de transport. Starea de
ntreinere a vnatului a fost precar, cu slbire accentuat. Examenul necropsic
general a fost urmat de examenul morfopatologic al ficatului. S-au prelevat
fragmente de esut hepatic care au fixate n formol 10%, apoi incluse n parafin,
secionate la 5 m i colorate prin metodele HEA, MGG, PAS. Examninarea i
microfotografierea s-au efectuat la MC5 cu oc. 10, ob. 10, 20, 40, i 63 Imm.

REZULTATE I DISCUII

Examenul histopatologic a relevat agresiuni recente asupra esutului


hepatic exprimate prin trasee hemoragice, angiocolit acut (fig. 1.), precum i
trasee hemoragico-necrotice, leziuni ale unor invazii anterioare exprimate prin
hepatit i angiocolit cronic (fig. 2., fig. 3.).

Fig. 1. Capr neagr. Ficat cu agresiune recent: hepatocite distruse, hemoragii,


traseu hemoragic, mrginit de infiltrat limfohistiocitar.
Col. MGG, oc. 10. x ob. 10

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Fig. 2. Capr neagr. Ficat cu agresiuni anterioare: trasee necrotice, rare hemoragii,
pigmeni hepatici, infliltrat eozinofilic. Col. MGG, oc. 10. x ob. 10

Fig. 3. Capr neagr. Ficat cu Cysticercus tenuicollis i angiocolit cronic. Reacia


conjunctiv este bogat, celulele macrofage au invadat peretele parazitului inducnd, n
cele din urm, moartea. Col. MGG, oc. 10. x ob. 20

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Pe msur ce procesul evolueaz intervine exsudaia sero-fibrinoas i


eozinofilic i ncep fenomene de hemoliz, fibrinoliz i resorbia componentei
lichide. Inflamaia eozinofilic este ntreinut printr-o reacie mezenchimal care
se instituie la periferia focarului, avnd ca punct de plecare fie capilarele
sinusoide fie mezenchimul perivascular din spaiile porte (4).
Aceste modificri sunt caracteristice hepatitei traumatice (hepatitis
cysticercosa), produs de Cysticercus tenuicollis (fig. 4.) n perioada migraiei
hepatice, extrem de agresiv, cauza exprimrii acute a metacestodozei. Invaziile
successive determin instalarea rezistenei i trecerea n forma cronic, dar cu
persistena leziunilor hepatice (1, 4).

Fig. 4. Capr neagr. Ficat cu Cysticercus tenuicollis n perioada dezvoltrii. Se


observ celule multinucleate n contact cu peretele parazitului i reacia conjunctiv cu
tendin de incapsulare. Col. MGG, oc. 10. x ob. 20

ntr-o faz avansat limfocitele sensibilizate i componentele


fosfolipidice care au aprut n timpul degradrii tisulare acioneaz asupra
macrofagelor transformndu-le n celule epitelioide. Detritusul necroctic bogat n
eozinofile intoxicate, constituie un corp strin pentru organism i induce
sinciializarea celulelor epitelioide sub forma unor complexe celulare mari, celule
gigante de corp strin care se dispun n coroan n jurul detritusului necrotic
(fig. 5.).
Cysticercus tenuicollis a fost identificat, n acelai timp, alturi de alte
structuri parazitare care au amplificat agresiunea multipl asupra esutului hepatic.

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Fig. 5. Capr neagr. Ficat cu Cysticercus tenuicollis mort, incapsulat, cu centrul


necrozat. Se observ celule gigante, de corp strin, dispuse n coroan, marginal, n
interiorul veziculei parazitare cu coninut necrozat. Reacia conjunctiv periferic este
bogat. Col. MGG, oc. 10. x ob. 20

Fig. 6. Capr neagr. Ficat parazitat concomitent i de alte structuri parazitare


neidentificate, alturi de Cysticercus tenuicollis. Col. MGG, oc. 10. x ob. 20

Cysticercus tenuicollis este metacestodul speciei Taenia hydatigena, care


paraziteaz ca adult n intestinul subire la cine, vulpe, lup, acal,. Cysticercus
tenuicollis paraziteaz animalele domestice (rumegtoare, porc, rareori la cal),

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animalele slbatice (mistre, cprior, capra neagr, muflon, cerb carpatin i mai
rar la cerbul loptar) i uneori omul. Determin cisticercoza seroaselor,
metacestodoz cunoscut n toate rile, cu pierderi economice importante, ca
urmare a nevalorificrii ficatului i a diminurii produciei de carne. Dup
migraia hepatic, C. tenuicollis traverseaz capsula Glisson i se fixeaz pe
seroasele abdominale, intrnd ntr-o faz de laten. Este o vezicul de culoare
albicioas, cu dimensiuni de 0,8-5 cm, monochistic, monocefalic, cu membrana
flasc. Conine lichid transparent i o pat opac, reprezentat de scolexul
viitorului cestod. Vezicula este ataat la seroase i suspendat de o prelungire
conjunctiv ca un pedicul (1, 2).
Carnivorele, gazd definitiv se infesteaz prin consumul viscerelor
provenite de la erbivore sau de la alte gazde intermediare. Carnivorele domestice
i slbatice rspndesc elementele parazitare (ou) odat cu fecalele, polund
spaiile de punare al erbivorelor. Infestarea erbivorelor se face pe cale digestiv
prin consumul vegetaiei sau i a apei contaminate cu ou de Taenia hydatigena
(5).

CONCLUZII

Examenul histopatologic al ficatului provenit de la exemplare de capr


neagr (Rupicapra rupicapra) a relevat agresiuni multiple acute i cronice asupra
esutului hepatic.
Suprapunerea evoluiei acute a agresiunilor peste agresiuni anterioare
cronice a contribuit la agravarea strii de boal cu depirea capacitii de
reactivitate a organismului i moartea animalelor.
Leziunile hepatice descrise s-au ncadrat n complexul patologic numit
hepatitis cisticercosa produs de Cysticercus tenuicollis n perioada migraiei
hepatice i a reactivitii celulare de corp strin.
Agresiunea asupra ficatului a fost multipl, exercitat i de alte structuri
parazitare neidentificate, alturi de Cysticercus tenuicollis.

BIBLIOGRAFIE

1. Dulceanu, N., Cristina Terinte, 1994 Parazitologie veterinar. Ed. Moldova, vol. II
th
2. Georgi, R., J., Marion E., Georgi, 1990 Parasitology for veterinarians. 5 edition,
Ed. W.B. Saunders Co. Philadephia, London, Toronto, Montreal, Sydney, Tokyo.
pag. 125-128
3. Nesterov, V., 1984 Bolile vnatului. Ed. Ceres, Bucureti, pag. 167-168.
4. Paul, I., 2001 Etiomorfopatologie veterinar, vol. III- Dismetabolii i parazitoze. Ed.
I.I. de la BradIai. pag. 195-197.
5. uteu, I., V. Cozma, 2004 Parazitologie clinic veterinar. Vol. I. Ed. Risoprint,
Cluj-Napoca, pag. 297-299.

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THE EFFECT OF ASCARIDIA GALLI INFESTATION


ON CHICKENS
M. S. ILIE*, I. COSOROAB,
Gh. DRBU, I. OPRESCU,
S. MORARIU, Narcisa RADBEA,
Alina ILIE, Florica MORARIU
U.S.A.M.V. Timioara

We carried out the infestation of 7 days old chickens through the


administration of earthworms taken from households affected by
ascaridiosis. Then we supervised the chicken weight and the water and
forage intake. The aim of this paperwork was to determine the losses caused
by the infestation with the nematode Ascaridia galli upon chicken
production. At the end of this study, we observed that earthworms play an
important role in transmitting Ascaridia galli in free range and that
chickens infested with Ascaridia galli gained an inferior body weight
compared to those not infested and kept under similar conditions.

Ascaridia galli is a common parasite in poultry, with a cosmopolite


distribution. The main host for this nematode is the domestic chicken (Gallus
gallus domesticus), but it was also found as parasite in turkey, pheasant, partridge,
goose and wild birds (13).
Ascaridiosis is one of the causes for the economic losses recorded within
the modern poultry breeding (2).
A study made in Denmark showed that Ascaridia galli prevalence, under
free range / organic conditions, was 100%, and under an intensive system on
permanent bedding 25% (9). Ascaridia galli is the most frequently met species of
parasite nematode in the traditional breeding from the South of Romania (7).
Natural infestation develops clinically in most of the cases and it produces rarely
losses caused by mortality (8). Ascaridia galli infestation is many times
associated to weight loss, the decrease of egg production, diarrhoea depending on
the parasitism intensity (8).
The aim of this paperwork was to identify losses caused by Ascaridia galli
infestation upon chicken production.

MATERIALS AND METHODS

Experimental animals
We purchased 32 one day old broilers (Flex Clasic hybrid from the
company Hubbard) from a company located in Bucov, Timi County. Chickens
were maintained on permanent bedding in a parasite free environment. Water

*
E-mail: marius.ilie@fmvt.ro

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administration and feeding with commercial forage containing anticoccidic


incorporated by the producer was made ad libitum. The recipe was prepared by
the company Agrobiotic on the base of a Biomin premix.
Experimental design
Up to the age of six days, chickens were bred together within a single
group, after that we randomized them into two groups. We constituted the
experimental groups, each with 16 chickens: C control; E in which we
administrated earthworms at the age of seven days.
We took earthworms from 3 intensely parasited households (Ticvaniul Mare
Cara-Severin county, Puuri Dolj county, erbneti Olt county). They were
administrated in two series, in tray, after a preceding diet of three hours, to group
E, cut into small pieces in order to maintain the possible parasitary elements
integer. They were alive at the administration moment to attract chickens with
their movements for eating.
We carried out vaccinations against Newcastle diseases in days 9 and 14
and against infectious bursitis in the 21st day of life. The microclimate was
assured according to the exploitation requirements for broilers bred on permanent
bedding.
After infestation, chickens were weighted in days 6, 10, 15, 20, 30, 40 52
and we supervised forage and water intake. Besides weighing, we carried out the
coproscopical examination of faeces.
The identification of nematode species was performed depending on the
determination keys (1, 4, 5, 12, 14).
Statistical method
The obtained experimental data were introduced in the software SPSS
version 7.5 and submitted to test t.

RESULTS AND DISCUTIONS

In the first experimental day, namely at the age of six days, chickens were
weighed, and the average weight obtained, in each group, had similar values, so
that the average weight in the infested group was 85.93 g, while in the control
group it was 84.68 g.
In days 5, 10, 15 after infection, the values of the average weights were also
similar, namely 112.5 g; 205.62 g; 366.78 g for the infested group, respectively
113.43 g; 210.93 g and 371.25 g for the control group.
The decrease of the average weight might be observed in the infested group
in day 25 after infestation 895.35 g, and in the control group it is 928 g. In days
35 and 47 the differences are important, so that the control group is heavier that
the infested one with about 91 g (E = 1533.92 g; C = 1625.33 g) in 34 days post
infection and with about 228 g in 47 days p.i. (E = 1993.07 g; C = 2221.53 g).
After the statistical analysis with the test t, we observed that there were
significant differences between the weights of the two groups, control C and
infested E. So, at the weighing performed in 15 days p.i., t value was 18.135 for
group C and 8.311 for group E, the significance being lower than 0.001 (p<0.001)

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at an interval of confidence of 95%. Similarly, data recorded at the weights within


days 35 and 47 p.i., significance p<0.001 between the groups C and E.
Table 1
Data obtained in the infested group E

CMZ CMZ Average


Age Exp. Forage Water Number
Forage water weight
(days) day (g) (ml) of chicken
(g) (ml) (g)
6 1 430 360 16 26.87 22.5 85.93
10 5 1185 680 16 74.06 42.5 112.5
15 10 805 860 16 50.31 53.75 205.62
20 15 590 1425 14 42.14 101.78 366.78
30 25 1795 4530 14 128.21 323.57 895.35
40 35 2640 5220 14 188.57 372.85 1533,92
52 47 2000 3250 13 153.8462 250 1993,07

Table 2
Data obtained in the control group C

CMZ Average
Age Exp. Forage Water Number of CMZ water
Forage weight
(days) day (g) (ml) chicken (ml)
(g) (g)
6 1 440 300 16 27.5 18.75 84.68
10 5 885 860 16 55.3125 53.75 113.43
15 10 785 960 16 49.0625 60 210.93
20 15 780 1550 16 48.75 96.875 371.25
30 25 1955 5400 15 130.3333 360 928
40 35 2640 5535 15 176 369 1625.33
52 47 2000 3400 15 133.3333 226.6667 2221.53

The results presented in the literature of speciality mention the fact that
there werent observed any significant differences until day 21 p.i., but after this
period, body weight has decreased significantly in the infested group. Weight
decrease became significant in the same time with the apparition of the first
clinical signs, around day 30 post infestation (2, 11).
The decrease of the body weight might due to intestinal mucosa disorders.
Glandular necrosis, within submucosa, induced by larva migration during tissue
phase and worms presence in the intestinal phase is the cause for adsorption
disorders and alteration of nutrients` metabolic processes at the level of the
intestinal wall (2).
Ascaridia galli infestation has a negative effect upon chicken weight. Some
authors specify that weight decrease is directly proportioned with the number of
ascarides within fowls intestine (10, 11).

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The number of the nematodes identified within the chickens` small


intestine, in group E, was different, so that we took a single individual from two
intestines, and from the others: two, four, nine, 10, 11, 12, respectively 13
individuals. Among the 13 digestive tracts, two were negative in terms of
Ascaridia galli presence and 11 were parasited.
Due to the fact that infestation was performed using earthworms, which can
ingest parasite eggs and transmit them to chicken, they represent an important risk
factor in transmitting Ascaridia galli infestation when they are consumed.
There are data for and against the earthworms role of paratenic host and
egg/larva transporter for A. galli in the literature of speciality (1, 3).

2400
2200
2000
1800
1600
1400
1200
1000
800
600
400
200
0
3 4 5 6 7
1 2
Age (week) Lot E Lot C

Fig. 1. Graphic representation of the experimental groups weight

CONCLUSIONS

Earthworms play an important role in transmitting Ascaridia galli under


traditional systems.
Chickens infested with Ascaridia galli had an inferior body weight
compared to those not infested and kept in similar conditions.

REFERENCES
nd
1. Anderson, R.C., 2000 - Nematodes parasites of vertebrates 2 Edition: their
development and transmission, CAB International - CABI Publishing.
2. Anwar H., Rahman, Y., 2002 - Effect of Ascaridia galli infestation on electrolytes and
vitamins in chickens. OnLine Journal of Biological Sciences 2 (10), 650-651.

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3. Augustine P.C., Lund E.E., 1974 - The fate of eggs and larvae of Ascaridia galli in
earthworms. Avian Diseases 3: 394-398.
4. Calnek, B.W., 1997 - Diseases of Poultry. Tenth Edition. Ed. Iowa State University
Press.
5. Eckert, J., Kutzer, E., Rommel, M., Burger, H.J., Korting, W., 1992 -
Veterinarmedizinische Parasitologie. Vol. IV. Auflage. Ed. Paul Parey, Berlin.
6. Ikeme M.M., 1971 - Weight changes in chickens placed on different levels of nutrition
and varying degrees of repeated dosage with Ascaridia galli eggs. Parasitology
63: 251-260.
7. Ilie, M.S., 2005 Helmintoze ale psrilor domestice crescute n sistem trediional n
unele judee din sudul Romniei, Scientia Parasitologica, 1-2, 84-88.
8. Juhl, J., Permin, A., 2002 The effect of Plasmodium gallinaceum on a challenge
infection with Ascaridia galli in chickens, Vet. Parasitol., 105, 11-19.
9. Permin, A., Bisgaard, M., Perman, M., Kold, J., Nansen, P., 1999 - Prevalence of
gastrointestinal helminthes in different poultry production system. British Poultry
Science, 40, 439-443.
10. Permin, A., Bojesen, M., Nansen, P., Bisgaard, M., Frandsen, F., Pearman
Margrethe, 1997 - Ascaridia galli populations in chickens following single
infections with different dose levels. Parasitol. Res. 83, 614 - 617.
11. Reid W.M., Carmon J.L., 1958 - Effects of numbers of Ascaridia galli in depressing
weight gains in chicks. Journal of Parasitology 44: 183-186.
12. Skryabin, K.I., 1991 - Key to Parasitic Nematodes, vol. 2, Oxyurata i Ascaridata,
Ed. E J. Brill.
13. Snabel, V., Permin, A., Magwisha, H.B., Suo, X., Varady, M., Tomasovicova, O.,
2001 - On the species identity of Ascaridia galli (Schrank, 1788) and Ascaridia
dissimilis (Perez Vigueras, 1931): a comparative genetic study, Helminthologia,
38, 4, 221-226.
14. Soulsby, E.J.L., 1982 - Helminths, Arthropods and Protozoa of Domesticated
Animals. 809 pp. Baillire Tindall. London.

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DINAMICA SEZONIER A POPULAIILOR DE IXODIDE


DIN DIFERITE ZONE GEOGRAFICE ALE ROMNIEI
SEASONAL DYNAMICS OF IXODIDAE POPULATIONS
IN DIFFERENT GEOGRAPHIC AREA FROM ROMANIA

Mariana IONI,
I.L. MITREA, M.C. BUZATU
U.S.A.M.V. Bucureti

The study about dynamics of ticks (Acari: Ixodidae) in three different


geographic areas from north-east and south, south-east of Romania, with
different ecological particularities, during 1999-2003 period, was
performed. Four species were identified: Ixodes ricinus, and Dermacentor
marginatus in all studied regions, while Rhipicephalus bursa was found
only in south area, and Hyalomma plumbeum plumbeum in south-east area.
The annual dynamics showed a seasonal activity with two different
peaks for Ixodes and Dermacentor: first one, in the spring and the begin of
the summer (June) and the second one in autumn (September, October). The
second peak represented 25,42-30% from the first one. The seasonal
activity of ticks populations from south and south-east areas started earlier
than those from north-east area.
The annual dynamics of the thermophile species - Rhipicephalus
bursa and Hyalomma plumbeum plumbeum, presented a seasonal activity
with only one peak, during of June.

Key words: ticks, dynamics, seasonal activity

Cpuele ixodide sunt artropode parazite temporar, a cror importan este


datorat att aciunii lor directe, ct i posibilitii transmiterii a numeroi ageni
patogeni (virusuri, rickettsii, parazii), rolul inoculator al cpuelor reprezentnd
un element de importan patogen i epidemiogen esenial (3, 11). Combaterea
eficient a ixodidelor se poate realiza doar pe baza cunoaterii exacte a
bioecologiei acestor acarieni, corelat cu particularitile geo-climatice ale ariilor
lor de rspndire (2, 10).
Abundena cpuelor n orice habitat este determinat de diveri factori,
cum ar fi covorul vegetal, clima - factori care afecteaz supravieuirea i
desfurarea fazelor care au loc n mediul extern (ponta, incubaia oulor,
ecloziunea, nprlirea), dar i de prezena gazdelor - pentru realizarea fazelor
parazite ale acestor artropode. Cunoaterea factorilor care determin densitatea
populaiilor de cpue are valoare predictiv, privitoare la consecinele anumitor
fenomene, cum ar fi modificri ale climei, modificarea raporturilor numerice ntre
diferite specii poteniale gazde (5, 8).

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Prezentul studiu a avut ca obiectiv prezentarea dinamicii sezoniere a


populaiilor de ixodide din diferite regiuni geografice ale Romniei, cu anumite
particulariti ecologice.

MATERIAL I METOD

Studiile au fost efectuate pe o perioad de 5 ani (1999-2003), n 3 zone


geografice diferite ale Romniei:
Judeul Suceava (n nord-est), caracterizat prin:
- clim temperat cu pronunat caracter continental, cu izoterme medii
anuale cuprinse ntre 8,4-9,28C i precipitaii anuale cuprinse ntre
418,3-706,2 mm;
- relief cu forme variate: muni, depresiuni intramontane, dealuri,
podiuri joase, lunci; vegetaie de pdure, rinoase i foioase, cu
arboret dezvoltat, pajiti naturale i secundare, cu flor dominat de
graminee;
Judeul Dmbovia (n sud):
- clim temperat, cu izoterme medii anuale de 10,57-11,21C i
precipitaii anuale cuprinse ntre 354,9-758,1 mm;
- relief dispus n trepte, de la nord la sud, cu: muni, zone subcarpatice,
depresiuni, cmpii, cu vegetaie de pdure (rinoase i fag n etajul
alpin, gorunete n zona subcarpatic i pduri de esen moale);
Judeul Teleorman (n sud):
- clim temperat continental de silvostep, cu izoterme medii anuale
cuprinse ntre 11,69-12,92C i precipitaii anuale cuprinse ntre
291,3-696,4 mm;
- relief cu cmpii uor nclinate, versani i lunci, cu altitudini de 50-130
m; vegetaie de pdure, cu strat arbustier foarte dezvoltat, adesea
compact, zvoaie (n luncile marilor ruri), tufriuri i pajiti de step;
Judeul Tulcea (n sud):
- clim temperat, cu slab caracter continental arid, cu izoterme medii
anuale cuprinse ntre 11,78-12,26C i precipitaii anuale de 419,5-
640,89 mm;
- relief variat: podiuri, dealuri, cmpii (cu o zon de step i una de
silvostep); vegetaie xerofil, pduri de lunc- zvoaie, cu arboret
dezvoltat i flor ierboas cu specii stepice, calcifile.
Colectarea cpuelor s-a realizat att din mediu metoda steagului
(Nutall), ct i de pe animale (manual, cu ajutorul unei pense) (6, 9, 10).
Cpuele recoltate au fost transportate la laborator n tuburi de sticl cu
alcool de 70; pentru identificarea speciilor au fost folosite chei de determinare (4,
7).

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REZULTATE I DISCUII

Analiza fenotipic a cpuelor colectate (9615 exemplare), a evideniat


prezena, ntr-o pondere variat, n zonele studiate (nord-est i sud-sud-est), a 4
specii de ixodide (Acari: Ixodidae):
- Dermacentor marginatus Sulzer 1776, cu o pondere de 37%,
identificat att n zonele din nord-estul Romniei, ct i n regiuni din
sud i sud-est;
- Ixodes ricinus Line 1758, cu pondere de 34,24%, prezent n toate
zonele de studiu;
- Rhipicephalus bursa Canestrini et Fanzago 1877, cu o pondere de
16,68%, identificat n zona de sud (judeul Dmbovia);
- Hyalomma plumbeum plumbeum Panzer 1795, cu o pondere de
12,06%, identificat n zona de sud-est (judeul Tulcea).
Pe baza datelor nregistrate (tabele 1-4) n perioada de studiu, au fost
ntocmite diagrame comparative ale dinamicii anuale a populaiilor de ixodide
identificate (figurile 1, 2).
Astfel, n cazul populaiilor de ixodide identificate n regiunile din nord-
estul rii (Ixodes ricinus i Dermacentor marginatus - specii cu valen
ecologic larg), s-au observat variaii interspecifice ale dinamicii lor anuale.
Debutul invaziei a fost nregistrat mai timpuriu la Dermacentor marginatus - n a
doua decad a lunii martie, fa de populaiile de Ixodes ricinus, care a prezentat
un debut al activitii parazitare constant, n ultima decad a lunii martie. De
asemenea, maxima indicilor invazionali a fost observat mai devreme la
Dermacentor spp. -sfritul lunii aprilie-nceputul lunii mai, fa de Ixodes - la
care valoarea maxim a indicilor invazionali a fost atins n ultima decad a lunii
mai i prima jumtate a lunii iunie.
La populaiile ambelor specii identificate n zona de nord-est, curba
dinamicii anuale a regresat n lunile de var, fr a atinge ns, punctul zero,
crescnd din nou toamna (septembrie-octombrie), amplitudinea medie a maximei
de toamn reprezentnd 25,42% din maxima de primvar - pentru Ixodes,
respectiv 17,25% pentru Dermacentor.
Aceleai diferene n dinamic au fost nregistrate i la populaiile din sud-
sud-estul rii ale celor dou specii, al cror areal se extinde la sud de izoterma
medie anual de 10. n aceste zone, debutul activitii parazitare a fost nregistrat
mai devreme fa de populaiile din nord-est, prima jumtate a lunii martie- la
ambele specii, cu un maxim al indicilor invazionali la sfritul lunii aprilie-prima
jumtate a lunii mai - pentru Dermacentor, respectiv a doua decad a lunii mai,
pentru Ixodes ricinus. Curba dinamicii a descris o regresie pronunat n lunile
iulie i august, n unele zone din sud chiar pn la zero. Atacul de toamn a avut
valori mai ridicate ale indicilor invazionali fa de cel din zonele de nord-est,
maxima de toamn reprezentnd 31,04% din maxima de primvar - la
Dermacentor, respectiv 30,47% - la Ixodes ricinus.

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50

40

30

20

10

0
I II III IV V VI VII VIII IX X XI XII

DmNE DmSE IxNE IxSE

Fig. 1. Reprezentarea grafic sintetic a dinamicii anuale a populaiilor de Ixodes ricinus


i Dermacentor marginatus n regiunile nord-estice i sud-sud-estice cercetate

60

50

40

30

20

10

0
I II III IV V VI VII VIII IX X XI XII

DmSE IxSE Rh.b. Hy.p.

Fig. 2. Reprezentarea grafic sintetic a dinamicii anuale a populaiilor de Ixodes ricinus,


Dermacentor marginatus, Rhipicephalus bursa i Hyalomma plumbeum plumbeum n
regiunile sud-sud-estice cercetate

Aceste diferene privind dinamica celor dou specii cu arii de rspndire


relativ asemntoare, cu un debut mai timpuriu pentru Dermacentor o maxim a
activitii parazitare atins mai devreme i o perioad mai restrns de parazitare

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fa de Ixodes, pot fi explicate pe baza exigenelor fa de umiditate pe care le


manifest Dermacentor marginatus; aceasta prefer biotopuri cu lstriuri i
flor iubitoare de umezeal, fiind ntlnit mai puin n regiunile sau n perioadele
uscate, aspecte semnalate i de ali autori (1, 4). Ixodes ricinus prefer pdurile de
foioase, fiind frecvent ntlnit n margini de pdure i zone defriate.
n cazul celorlalte dou specii identificate doar n zonele din sud-sud-est
Rhipicephalus bursa i Hyalomma plumbeum plumbeum - specii termofile,
diagrama dinamicii anuale a descris un singur peak evident - n luna iunie
pentru ambele specii, peak-ul de toamn, atunci cnd a fost prezent (la
Rhipicephalus bursa) a reprezentat n medie doar 10% din maxima de primvar.

CONCLUZII

1. n perioada 1999-2003 au fost efectuate observaii privind dinamica


ixodidelor n 3 zone geografice ale Romniei, cu particulariti ecologice diferite:
judeul Suceava (nord-est), judeele Dmbovia i Teleorman (n sud) i Tulcea (n
sud-est).
2. Analiza fenotipic a cpuelor colectate (9615 exemplare), a evideniat
prezena a 4 specii aparinnd familiei Ixodidae (Acari): Dermacentor marginatus
(pondere 37%), Ixodes ricinus (34,24%), identificate n toate zonele de studiu;
Rhipicephalus bursa (16,68%), n zona de sud; Hyalomma plumbeum plumbeum
(12,06%), identificat n zona de sud-est.
3. Dinamica anual a speciilor de ixodide identificate n zonele de studiu a
evideniat diferene n momentele de debut i, respectiv, parazitism maxim la
animale, n funcie de arealul de distribuie i de evoluia anual a principalilor
factori ecologici (temperatur, umiditate, vegetaie).
4. La populaiile din regiunile sud-sud-estice, debutul i maximul activitii
parazitare a fost mai timpuriu fa de cele din zonele nord-estice; maxima de
toamn a avut valori medii de peste 30% din maxima de primvar, fa de
speciile din nord-est la care maxima de toamn a reprezentat 25,42% din cea de
primvar - pentru Ixodes, respectiv 17,25% pentru Dermacentor.
5. La speciile termofile - Rhipicephalus bursa i Hyalomma plumbeum
plumbeum, identificate numai n regiunile sudice i sud-estice, curba dinamicii
anuale a descris un singur vrf evident, nregistrat n luna iunie, pentru ambele
specii.

BIBLIOGRAFIE

1. Bdescu C., 1969 - Contribuii la studiul ecologiei cpuelor din familia Ixodidae
Murray 1877, din differite zone ale Romniei, Comunicri tiinifice, Editura
Didactic i Pedagogic, Bucureti, 381-394
2. Bourdeau P., 1993 - Les tiques dimportance Vtrinaire et mdicale. Le Point
Vtrinaire, 25 (151): 13-42
3. Cosoroab I., 2000 - Acarologie veterinar, Editura Mirton, Timioara

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4. Feider Z., 1965 - Fauna R.P.R. Arachnida, vol V, Fasc.2, Editura Academiei
5. Gray J.S., 1991 - The development and seasonal activity of the tick, Ixodes ricinus: a
vector of Lyme borreliosis. Rev Med Vet Entomol 79: 323-333.
6. Ioni Mariana, Mitrea L.I., Moise G., Gin D., Onofrei O., Marinescu C., 2001 -
Influena factorilor ecologici asupra structurii fenotipice i dinamicii sezoniere a
populaiilor de Ixodidae n unele regiuni ale Romniei. Simpozion la 140 de ani,
Alma Mater Veterinaria Bucurescensis, 192
7. Keirans J.E., Robbins R.G., 1999 - A world checklist of genera, subgenera and
species of ticks (Acari: Ixodidae). Published from 1973-1997. J. Vector Ecol., 24:
15-129
8. Lindgren E, Tlleklint L, Polfeldt T., 2000 - Impact of climatic change on the
northern latitude limit and population density of the disease-transmitting
European tick Ixodes ricinus. Environ Health Persp 108: 119-123.
9. Mitrea L.I., Ioni Mariana, Moise G., Gin D., Onofrei O., 2001 - Aspecte privind
infestaiile cu cpue Ixodidae la animale n regiuni din N i S-E Romniei,
Lucrri tiinifice, USAMV, Iai, vol. 44 (3), fasc. II: 523-529
10. Sonenshine D.E., 1993 - Biology of ticks. Oxford, Oxford University Press, vol. 2
11. uteu I., N. Dulceanu, 2001 - Parazitozele cutanate la animale, Editura Risoprint,
Cluj-Napoca

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TESTAREA EFICACITII MOXIDECTINEI


N HELMINTOZE INTESTINALE LA GALINACEE
THE EFFICACY OF MOXIDECTINE IN INTESTINAL HELMINTOSIS
OF GALLINACEAE

C. MAGDA, Daniela MOTOGNA,


V. COZMA

The aim of our study was to test the efficacy of moxidectine in


inthestinal helmintosis of Gallinaceae. The researches performed during
June 2005, on birds bred in exthensive system, from Bistria Nsud
county. We have used Cydectin 1%, applied by subcutaneus injection, 0.3
mg/bw. The results showed that moxidectine had a good efficacy after 7
days, in ascaridiosis/heterakidosis, capilariosis and strongyloidosis.

INTRODUCERE

Introducerea avermectinelor pentru uzul veterinar a reprezentat un


eveniment major n terapia i profilaxia multor boli parazitare (Ciser et Galiano,
1997). Aceasta s-a datorat att eficacitii antiparazitare la un dozaj redus (0,2-
0,5mg/kc), ct i spectrului terapeutic larg extins asupra nematodelor,
arachnidelor i insectelor (Campbell, 1989).
Milbemicina, descoperit n 1974, este o lacton macrociclic obinut prin
fermentaia miceilor din genul Streptomyces. Moxidectina, este un derivat obinut
prin modificri chimice ale nemadectinei, produsul obinut prin fermentaia
micetului Streptomyces cyanogriseus. Diferena ntre milbemicine i avermectine
o constituie prezena unui dizaharid, substituient al carbonului 13, prezent n
avermectine i absent n milbemicine (William, 1999). Dei proprietile acaricide
ale milbemicinei au fost recunoscute iniial, eficacitatea antihelmintic a acesteia
a fost demonstrat ulterior descoperirii avermectinelor, neexistnd diferene n
ceea ce privete modul lor de aciune (Fisher et al, 1990).

MATERIAL I METOD

Cercetrile privind efectul moxidectinei asupra helmintozelor intestinale la


galinacee au fost efectuate n luna iunie 2005, pe psri crescute n sistem
gospodresc, n judeul Bistria-Nsud. Probele coproparazitologice au fost
prelucrate i examinate n cadrul disciplinei de Boli Parazitale a Facultii de
Medicin Veterinar, Cluj-Napoca.
Galinaceele luate n studiu, au fost lotizate astfel:

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-
Lotul I (60 psri) tratate cu moxidectin (Cydectin 1%) n doz de
0,3 mg/kg corp, s.c., o administrare;
- Lotul II (60 psri) - tratate cu moxidectin (Cydectin 1%) n doz de
0,3 mg/kg corp, s.c., dou administrri, la interval de 7 zile;
- Lotul III (60 psri) lot martor pozitiv, netratat.
n vederea diagnosticrii infestaiilor parazitare, ante-terapeutic s-au
recoltat probe coproparazitologice de la toate trei loturile. Probele au fost
prelucrate prin metoda Willis, sedimentare activ, i McMaster pentru
evidenierea intensivitii acestora (OPG).
Recoltrile de fecale au fost efectuate n ziua 0, 7 i 14 al experimentului.

REZULTATE I DISCUII

Rezultatele examenelor coproscopice efectuate ante-terapeutic la cele 3


loturi de psri luate n studiu i intensivitatea parazitismului, sunt prezentate n
tabelul 1.
Profilul endoparazitar ante-terapeutic n cazul lotului I a fost reprezentat de
infestaii parazitare cu: Ascaridia / Heterakis, cu o medie a intensivitii de 150
OPG, Capillaria spp. cu o medie a intensivitii de 200 OPG i Strongyloides
avium, cu o medie a intensivitii de 50 OPG.
Tabelul 1
Valoarea intensivitii infestaiilor endoparazitare la galinacee, ante-terapeutic

Strongylo
Lot/ Eimeria O.P.G. Ascaridia/ O.P.G. Capillaria O.P.G. O.P.G.
ides
Specie spp. (X) Heterakis (X) spp. (X) (X)
avium
Lot I. - - + 150 + 200 + 50
Lot II. + 100 + 50 + 550 - -
Lot III. + 50 + 100 + 350 - -

La lotul II s-au diagnosticat infecii cu Eimeria spp., cu media intensivitii


de 100 OPG, infestaii cu Ascaridia/Heterakis, cu o medie a intensivitii de 50
OPG i Capillaria spp. cu o medie de 550 OPG.
n cazul lotului III, martor netratat, s-au diagnosticat infecii cu Eimeria
spp. cu media intensivitii de 50 OPG, infestaii cu Ascaridia/Heterakis cu media
intensivitii de 100 OPG i Capillaria spp. cu media intensivitii de 350 OPG.
Ante-terapeutic, valoarea cea mai ridicat a intensivitii infestaiei helmintice, a
fost diagnosticat n cazul lotului II, la Capillaria spp, cu 550 OPG (fig. 1). n
urma unui studiu epidemiologic efectuat n judee din sudul trii, Ilie (2005)
semnaleaza la gini din sistem gospodresc, infestaii cu Ascaridia galli,
Heterakis gallinae, Capillaria spp., Raillietina spp., Gongylonema ingluvicola i
Cheilospirura hamulosa.

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600 550
500

400 350
O.P.G.

300
200
200 150
100 100
100 50 50 50
0 0 0
0
Lotul I Lotul II Lotul III

Eimeria spp. Ascaridia/Heterakis Capillaria spp. Strongylloides

Fig. 1 - Valoarea intensivitii infestaiilor endoparazitare la galinacee,


ante-terapeutic (OPG)

Rezultatele obinute n urma examenelor coproparazitologice efectuate la


loturile I, II, i III, post-terapeutic la 7 zile dup prima administrare a
moxidectinului sunt prezentate n tabelul 2.
Tabelul 2
Valoarea intensivitii infestaiilor endoparazitare la galinacee,
la 7 zile post-terapeutic

Strongylo
Lot/ Eimeria O.P.G. Ascaridia/ O.P.G. Capillaria O.P.G. O.P.G.
ides
Specie spp (X) Heterakis (X) spp. (X) (X)
avium
Lot I. - - - - - - - -
Lot II. + 50 - - - - - -
Lot III. + 100 + 100 + 400 - -

n cazul loturilor I i II se observ dispariia infestaiilor helmintice la 7 zile


post-terapeutic, evideniindu-se doar infecii cu Eimeria spp. cu o medie a
intensivitii de 50 OPG.
n cazul lotului III, netratat, n ziua a 7-a a experimentului, au fost
diagnosticate infecii cu Eimeria spp. cu o medie a intensivitii de 100 OPG,
observndu-se persistena infestaiei helmintice cu Ascaridia/Heterakis, cu 100
OPG, identic cu cea constatat la controlul precedent i creterea mediei
intensivitii n cazul infestaiei cu Capillaria spp., la 400 OPG (fig. 2).

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400
400
350
300
250
O.P.G.

200
150
100 100
100
50
50
0 0 0 0 0 0 0 0
0
Lotul I Lotul II Lotul III

Eimeria spp. Ascaridia/Heterakis Capillaria spp. Strongylloides

Fig. 2 - Valorea intensivitii infestaiilor endoparazitare la galinacee, la 7 zile post-


terapeutic (OPG)

Rezultatele obinute la 14 zile post-terapeutic n cazul lotului I, respectiv la


7 zile de la cea de a doua administrare a moxidectinului la lotul II, ct i la lotul
martor netratat, sunt redate n tabelul 3.
Tabelul 3
Valorea intensivitii infestaiilor endoparazitare la galinacee,
la 14 zile post-terapeutic

Strongylo
Lot/ Eimeria OPG Ascaridia / OPG Capillaria OPG OPG
ides
Specie spp (X) Heterakis (X) spp. (X) (X)
avium
Lot I. - - - - - - - -
Lot II. + 50 - - - - - -
Lot III. + 50 + 50 + 600 - -

600
600

500

400
O.P.G.

300

200

100 50 50 50
0 0 0 0 0 0 0 0
0
Lotul I Lotul II Lotul III

Eimeria spp. Ascaridia/Heterakis Capillaria spp. Strongylloides

Fig. 3 - Valorea intensivitii infestaiilor endoparazitare la galinacee,


la 14 zile post-terapeutic (OPG)

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n cazul loturilor tratate cu moxidectin, n ziua a 14-a a tratamentului, se


observ o situaie identic cu cea precedent, respectiv lipsa infestaiilor
helmintice. La lotul martor netratat s-au meninut infecia cu Eimeria spp. i
infestaiile cu Ascaridia/Heterakis (OPG=50) i Capillaria spp. (OPG=600)
(fig. 3).
Melhorn i colab. citat de uteu i Cozma (2004) recomand n lipsa
antihelminticelor moderne, n capilarioz la psri, avermectine n doz de
0,2mg/kg corp, s.c.. De asemenea uteu i Cozma (2004) recomand n
amidostomoz i singamoz la psri avermectine n doz de 0,2 mg/kg, s.c..
Fenomene de toxicitate n cazul utilizrii ivermectinei la psri apar la dozajul de
5,4 mg/kg mas corporal, manifestate printr-o somnolen ce dureaz circa 4 ore,
n timp ce doza de 48,6 mg/kg mas corporal a determinat moartea psrilor
dupa 5 ore p.i. (Zeman, 1987). Coles (1987) recomand n cazul psrilor
administrarea oral a avermectinelor, considernd eficacitatea acesteia similar cu
cea pe cale injectabil.
Mufit i colab. au tratat cu succes ria cnemidocoptic la perui, prin
aplicri spot-on de moxidectin (Cydectin Injection for cattle), n doz de
1mg/pasre (0,1 ml Cydectin), fr a nregistra fenomene de intoxicaie, dei doza
administrat a fost de circa 25-29 mg/kc (Mufit et al, 1999).
Experimentri de neurofiziologie arat faptul c lactonele macrociclice
blocheaz ireversibil transmisia neuromuscular, acionnd la nivelul sinapselor,
stimulnd producerea acidului gama-aminobutiric (GABA), care este inhibitor al
stimulilor nervoi (uteu 1995). Stimularea producerii de GABA presinaptic,
modific biochimismul receptorilor postsinaptici, neuronii motori rmn ncrcai
negativ, astfel nct stimulii excito- i inhibitori nu mai pot fi receptai de celulele
musculare i astfel se instaleaz paralizia (Campbel i colab. 1979). n plus
acestea intervin direct asupra ionilor de clor, independent de prima relaie. Ionii
de clor ptrund n celula postsinaptic, aceasta avnd sarcin negativ, nu mai
primete semnale prin neuronii motori, producndu-se paralizia. Nu toate formele
evolutive ale unei specii sunt deopotriv receptive la aciunea avermectinelor, iar
modul de hrnire influeneaz esenial vulnerabilitatea acestora.

CONCLUZII

Cercetrile efectuate n iunie 2005, pe un efectiv de 180 de psri, crescute


n sistem gospodresc, n judeul Bistria-Nsud, au relevat urmtoarele:
- Moxidectina (Cydectin 1%), administrat s.c., n doz de 0,3 mg/ kg
greutate corporal, a avut eficacitate maxim la 7 zile post-terapeutic
in ascaridioz/heterakidoz, capilarioz i strongiloidoz.
- n cazul lotului netratat, infestaiile helmintice s-au meninut pe tot
parcursul experimentului.

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BIBLIOGRAFIE

1. Campbell W., 1989 Ivermectin and abamectin, Merk Institute for Therapeutic
Research, 216-227
2. Campbell W., Blair S., Lotti J. 1979 J. Helminthology, 53, 254-256
3. Ciser L., Galiano I., 1997 - Indexul produselor de uz veterinar din import, Edit. Ceres
Bucureti
4. Coles B.H., 1987 - Ivermectin for birds, vet. Rec., 120:604
5. Fisher M.H., Mrozic H., 1990 Ivermectin and abamectin, Campbel WC, ed.,
Berlin/Heidelberg, Springer-Verlag
6. Ilie M.S., 2005 Helmintoze ale psrilor domestice crescute n sistem tradiional n
unele judee din sudul Romaniei, Rev. Scientia Parasitologica, vol 6 nr 1-2, 84-88
7. Mufit T., Erkut T., Aysen G., Aynur G., 1999 Therapy of Knemidocoptic mange in
budgerigars with spot-on application of Moxidectin, Tr. J. Of Veterinary and
Animal Science, 23, 173-174
8. uteu E. 1995 Utilizarea avermectinelor n terapia parazitozelor la animale, Rev.
Rom. Med. Vet., vol 5, nr 4, 344-354
9. uteu E., Cozma V., 2004 Parazitologie clinic veterinar, Edit. Risoprint, Cluj-
Napoca.
10. Zeman P., 1987 Systemic efficacy of ivermectin against Dermanyssus gallinae (De
Geer, 1778) in fowls, Vet. Paras., 23:141-146
11. William G.R., 1999 Ivermectin / Doramectin /Moxidectin - Structure and generation,
A scientific review, Veterinary Bulletin Merial

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ISOSPOROZA CARNIVORELOR DOMESTICE:


ASPECTE TERAPEUTICE I CLINIC-EVOLUTIVE
THERAPEUTICAL AND CLINCAL ASPECTS
IN ISOSPOROSIS OF DOMESTIC CARNIVOROUS

I.L. MITREA, Violeta ENCHESCU,


Mariana IONI, M.C. BUZATU
U.S.A.M.V. Bucureti

The therapeutical and clinical evolution in isosporosis of dogs and


cats was registered. The studies included 21 animals (13 dogs and 8 cats)
with clinical signs due of Isospora (Cystoisospora) infections and confirmed
by coproparasitological examinations.
The treatment included specific medication (sulphamides, triazones
(toltrazuril) and amprolium) in association with diet and symptomatic
treatment.
The clinical amelioration was registered beginning in the second day
after treatment with normal consistence of feces in 3-4 days. In some cases,
the reappearance of clinical signs after 10 days was registered. The second
treatment was salutary.
After the treatment, the coproparasitological exams were negative.

Key words: isosporosis, dog, cat, therapy, clinical,


coproparasitological examination

Isosporoza este o coccidioz produs de Isospora (Cystoisospora) spp.,


protozoare ncadrate n genul Eimeria, familia Eimeridae, ordinul
Eucoccidiida/Eucoccidiorida, subclasa Coccidia (Coccidiasina), clasa Sporozoa
(Apicomplexa) (8, 9, 12). La cine se ntlnesc patru specii de Cystoisospora: C.
canis, C. ohioensis, C. burrowsi i C. neorivolta. Aspectul microscopic al
oochisturilor nu permite diferenierea dup morfologie sau mrime a speciilor C.
ohioensis, C. burrowsi i C. neorivolta care sunt uneori regrupate ntr-o singur
entitate: complexul C. ohioensis (1, 3, 4, 7). La pisic paraziteaz Cystoisospora
(Isospora) felis (foarte des identificat n fecalele pisicilor domestice) i Isospora
rivolta (6, 14).
Dei sunt unele dintre cele mai rspndite boli parazitare ale animalelor de
companie, coccidiozele, n spe isosporozele, nu sunt diagnosticate n mod
curent, fiind adesea neglijate, chiar total ignorate sau confundate cu alte boli
predominant digestive, datorit evoluiei de multe ori insidioase (13). Totui,
isosporoza este o ameninare serioas pentru sntatea carnivorelor domestice.
Afectnd preponderent animalele tinere, mai ales n perioada nrcrii, boala
evolueaz ndeosebi la animalele crescute n colectiviti cu condiii igienice
necorespunztoare, unde determin pierderi nsemnate. Astfel, exist cazuri severe

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cnd se poate produce o enterit cataral-hemoragic cu diaree, anemie, emaciere


rapid, depresie i slbiciune, uneori ameninnd chiar viaa animalului (16). n
general isosporoza nu este considerat o cauz important a diareei la animalele
adulte. De multe ori infecia este asimptomatic chiar i la animale tinere.
Infeciile eimeriene, inclusiv cele cu Isospora constituie, ns i un factor
favorizant pentru intervenia altor ageni patogeni i evoluia unor maladii asociate.
Numrul speciilor de Isospora care sunt patogene pentru cini i pisici este
controversat (5). Identificarea oochisturilor n fecalele animalelor sntoase indic
o infecie comensal autolimitant, care rareori necesit tratament (17), dei
acesta poate ajuta la reducerea contaminrii mediului, scurtnd perioada patent
i durata bolii. Dac semnele clinice sunt atribuite unei coccidioze, trebuie
instituit un tratament cu anticoccidiene eficiente (10, 11, 12).
Tratamentul isosporozei carnivorelor include un tratament specific, cruia i
se asociaz i unul simptomatic. Astfel, concomitent cu tratamentul specific se
intervine pentru rehidratarea animalelor cu diferite soluii rehidratante, precum i
administrarea de vitamine, antidiareice i antihemoragice (15). Se trateaz
anemia. Se recomand i un regim alimentar dietetic.

MATERIAL I METOD

Pentru determinarea eficacitii terapeutice i a evoluiei clinice a


isosporozei la carnivore, n urma tratamentului, au fost luai n studiu 21 pacieni
(13 cini i 8 pisici) examinai n cabinete veterinare din Bucureti, diagnosticul
confirmndu-se n cadrul laboratorului de Parazitologie-Boli parazitare al
Facultii de Medicin Veterinar, Bucureti.
Cazurile studiate au fost alese dintre pacieni ce prezentau semne clinice
atribuite isosporozei, confirmate prin examen de laborator.
Suspiciunea de isosporoz a avut la baz coroborarea semnelor clinice
(diaree cu un aspect necaracteristic, eventual cu prezena de mucus i snge, cu
evoluie n general acut i neacompaniat de febr, cu sau fr modificarea
strii generale), cu datele epidemiologice (animale de vrst tnr sub 6 luni,
provenite, n general, din colectiviti). n cazul unei colectiviti, boala se
prezenta ca o enzootie de enterite, uneori hemoragice, afectnd de obicei numai
tineretul, frecvent n perioada din preajma nrcrii sau la cteva zile dup
aciunea unui factor stresant (transport, schimbarea locuinei etc.); ntrzierile n
cretere au afectat inegal indivizii de la aceiai ftare.
Diagnosticul de laborator a fost stabilit n urma examenului microscopic al
fecalelor, prin depistarea oochisturilor specifice [2]. S-a folosit o metod
ovoscopic de concentrare - metoda de flotaie (Willis), folosind soluia
suprasaturat de clorur de sodiu.
Terapia celor 21 de pacieni s-a efectuat cu diferite substane, singure sau
n asociere, conform tabelului 1. Au fost ncercate formule terapeutice variate, de
la medicaie pe cale oral, medicaie injectabil sau asocieri ntre acestea.

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Tabelul 1
Tratamente utilizate la carnivore cu isosporoz

Posologie
Medicamente utilizate
Durata
Doza i calea de administrare
tratamentului
Sulfamethoxidiazina 1 2ml/5kg corp/zi s.c., (doza
3 pn la 6 zile
sol. 20 % (Ultrasol ) initiala), apoi 0,5ml/5kg/zi
Combinaia sulfamerazin,
sulfadiazin, sulfatiazol cu
1ml/5 kg s.c. 4 5 zile
trimetroprim (Trimethosulf ) soluie
injectabil
Combinatia sulfametoxazol 200mg

trimetroprim 40mg (Alfatrim soluie 1ml/20 kg s.c. 5 zile
injectabila)
Sulfadimidina trimetroprim 20 mg sulfadimidina i 5 mg
5 zile
(Sultrim suspensie buvabila) trimetroprim/kg corp, per os.
Scourban Plus
(sulfadimidina, sulfaguanidina,
sulfadiazina, streptomicina,
neomicina, hioscina, clorura de 0,5 1 ml/1 kg corp de 2 ori/zi 3 5 zile
sodiu, gluconat de calciu, sulfat de
magneziu, clorura de potasiu,
caolin, pectina, glicina)
O singura
Toltrazuril (Baycox) soluie orala 30 mg/kg corp la pisica administrare,
2,5% 20 mg/kg corp la cine repetare la 10
14 zile
Amprolium (Amprolium 20%) 100 mg amprolium/animal/zi 7 zile
Furazolidon 10 mg/kg corp, de 2 ori/zi 5 zile

Galiprotect (oxitetraciclina,
furazolidona, vitamine, magneziu, 1 - 2 cp./kg corp/zi 3 zile
zinc, metionina)

n majoritatea cazurilor tratamentului medicamentos i-a fost asociat, dup


caz, un tratament simptomatic, tratament de susinere a organismului, tratament
igieno-dietetic i alte tratamente endoparazitare (n cazuri de poliparazitism).

REZULTATE I DISCUII

Aspectele clinice consemnate consecutiv tratamentelor efectuate, sunt


prezentate n tabelele 2 i 3.

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Tabelul 2
Rezultate obinute n terapia isosporozei la cine

Nr.
Medicaia utilizat Evoluia semnelor clinice dup tratament
cazuri
La 48 h p.t. s-a observat o ameliorare a
semnelor clinice. Semnele clinice au
Sulfamethoxidiazin
1 reaprut la o sptmn de la ncetarea
sol. 20 % (Ultrasol )
tratamentului s-a impus reluarea
tratamentului.
Suspensie antidiareic - Dup 3 zile de tratament s-a nregistrat o

Scourban Plus (sulfadimidin, recuperare total, la un caz, fie ameliorarea
sulfaguanidin, sulfadiazin, vizibil, la cellalt caz, n funcie de
streptomicin, neomicin, gravitatea iniial a semnelor clinice. La 10
2
hioscin, clorur de sodiu, zile dup ncheierea tratamentului,
gluconat de calciu, sulfat de examenul coproparazitologic a fost pozitiv
magneziu, clorur de potasiu, (la unul din cazuri) repetarea
caolin, pectin, glicin) tratamentului.
Dup 2 sptmni, la un caz s-a constatat
reapariia diareei i a fost reluat tratamentul.
Amprolium 2
Dup o lun, examenul coproparazitologic a
fost negativ.
A 2-a zi de la nceperea tratamentului, s-a
constatat o ameliorare a strii pacientului
(recuperarea apetitului, scaun de
consisten mai tare), iar n a 3-a i a 4-a zi,
Toltrazuril 2
scaunul era normal. La unul din cazuri s-a
constatat reapariia diareei, la un interval de
2 sptmni, cnd a fost repetat
tratamentul.
Combinaia sulfamerazin, Dup 2 zile de tratament: ameliorare vizibil
sulfadiazin, sulfatiazol cu (dispariia vomei, fecale pstoase).
1
trimetroprim (Trimethosulf) Examenul coproparazitologic la o
soluie injectabil sptmn negativ.
Sulfamerazin, sulfadiazin, Recuperare complet n 3-5 zile; examenul
sulfatiazol cu trimetroprim + 1 coproparazitologic la o sptmn
suspensie antidiareic negativ.
Galiprotect(oxitetraciclina,
furazolidona, vitamine, magneziu, 1 Diareea a ncetat din a 4-a zi de tratament.
zinc, metionina)
Diareea s-a redus din a 2-a zi i a disprut
n a 4-a zi. Apetitul a revenit la normal n a
Sulfadimidina trimetroprim
2 3-a zi. Diareea a reaprut, la un caz, la o
(Sultrim suspensie buvabila)
sptmn de la terminarea tratamentului
reluarea terapiei.
Combinatia sulfametoxazol
200mg trimetroprim 40mg 1 n 5 zile pacientul era refcut complet.
(Alfatrim soluie injectabil)

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Tabelul 3
Rezultate obinute n terapia isosporozei la pisic

Nr.
Medicaia utilizat Evoluia semnelor clinice dup tratament
cazuri
Ameliorarea semnelor clinice dup 2 3 zile
de tratament, recuperare complet dup 5
Sulfamethoxidiazina
2 6 zile de tratament; la un caz, diareea a
sol. 20 % (Ultrasol )
reaprut dup 12 zile repetarea
tratamentului
Suspensie antidiareic - Scourban

Plus (sulfadimidin,
sulfaguanidin, sulfadiazin, Dup 3 zile s-a nregistrat ameliorarea
streptomicin, neomicin, semnelor clinice i scaun de consisten
2
hioscin, clorur de sodiu, pstoas. Dup 6 zile examenul
gluconat de calciu, sulfat de coproparazitologic a fost negativ.
magneziu, clorur de potasiu,
caolin, pectin, glicin)
Sulfamethoxidiazin + suspensie Scaunul a devenit de consisten normal
1
antidiareic din a 2-a zi de tratament.
Scaun normal i apetit refcut n 2 3 zile.
Toltrazuril 2 La un caz, diareea a reaprut dup 2
sptmni reluarea tratamentului.
Absena semnelor clinice, dar examen
Furazolidon 1 coproparazitologic pozitiv, la o sptmn
de la nceperea tratamentului.

Analiza comparativ a tratamentelor efectuate la cazurile diagnosticate ne-a


permis unele evaluri:
- Rezultate bune s-au obinut prin folosirea toltrazurilului (Baycox) soluie
2,5% n doz de 30 mg/kg la pisoi i 20 mg/kg la cei; apetitul a revenit la
normal chiar de a doua zi dup instituirea tratamentului, iar scaunul a revenit la
consistena normal n 2 zile la pisoi, respectiv 3 - 4 zile la ceii tratai cu
toltrazuril.
- S-a remarcat eficiena produsului Scourban Plus care reprezint o asociere
de 3 sulfamide (sulfadimidin, sulfaguanidin, sulfadiazin), 2 antibiotice eficiente
pe tubul digestiv (streptomicin, neomicin) i alte compo-nente (hioscin, clorur
de sodiu, gluconat de calciu, sulfat de magneziu, clorur de potasiu, caolin, pectin,
glicin). Produsul se prezint sub form de suspensie buvabil avnd efect
anticoccidian i n acelai timp un spectru larg antibacterian, contribuie la
restabilirea echilibrului electrolitic i reduce motilitatea intestinal. Prin tratamentul
cu Scourban Plus s-a obinut o ameliorare vizibil a semnelor clinice dup 2 zile de
tratament, pacientul fiind complet refcut dup 5 zile de tratament, cu dispariia
oochisturilor din fecale. n alte cazuri, fecalele i-au recptat consistena normal
n 3 zile - 5 zile, n funcie de gravitatea semnelor clinice.

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- Au existat cazuri (5 cei din 13, respectiv 2 pisoi din 8), n care semnele
clinice i eliminarea de oochisturi au reaprut la 1 - 2 sptmni dup tratamentul
cu anticoccidiene. n aceste cazuri, tratamentul a fost repetat, o nou reut
nemaifiind nregistrat. Repetarea tratamentului dup 10 zile a avut efect salutar.
Durata de evoluie a bolii la animalele tratate a fost n medie de 3 - 5 zile,
mbuntiri ale strii clinice observndu-se dup 2 - 3 zile de tratament. Dup 5 -
6 zile de tratament, n majoritatea cazurilor nu au mai fost identificate oochisturi
n fecale sau numrul acestora a sczut dramatic. S-au nregistrat totui, remisiuni
ale semnelor clinice la 1 - 2 sptmni de la ncheierea tratamentului, asociate cu
reapariia oochisturilor n fecale, la unele animale tratate cu sulfamide i
amprolium, dar i ntr-un caz n care tratamentul s-a efectuat cu toltrazuril. Situaii
asemntoare au fost relatate i de ali autori [10, 11]. Revenirea semnelor clinice
ar putea fi pus pe seama unei aciuni coccidiostatice sau coccidiocide imperfecte
a medicamentelor, unei contaminri intense a adposturilor, ineficienei
dezinfectantelor folosite mpotriva oochisturilor sau sporulrii rapide a acestora.
Reinfectarea pare s nu se mai produc dup un al 2-lea tratament cu
anticoccidiene, dar aici pot interveni i creterea vrstei sau imunitatea
protectoare, care pot limita infecia sau pot extinde perioada prepatent. La
revenirea semnelor clinice pot contribui i posibilele infecii tisulare latente.

CONCLUZII

Studiul a fost realizat pe 13 cini i 8 pisici, avnd drept scop evaluarea


terapiei cu diverse medicamente n isosporoza carnivorelor. Tratamentele s-au
efectuat n principal cu sulfamide, triazone (toltrazuril) i amprolium. Msurile
igienodietetice i tratamentul simptomatic au fost ntotdeauna asociate celui
specific antiparazitar.
Rezultate terapeutice bune s-au obinut prin folosirea toltrazurilului; de
asemenea, folosirea pe cale oral a unei suspensii antidiareice ce conine o
combinaie de 3 sulfamide, 2 antibiotice specifice tubului digestiv, electrolii i
substane antidiareice (Scourban Plus) s-a dovedit eficient att pentru scurtarea
evoluiei bolii, ct i pentru reducerea numrului sau chiar oprirea eliminrii
oochisturilor prin fecale.
Durata de evoluie a bolii la animalele tratate a fost n medie de 3 5 zile,
mbuntiri ale strii clinice observndu-se dup 2 3 zile de tratament.
Tratamentul a condus la dispariia oochisturilor din fecale sau micorarea drastic
a numrului acestora.
La unele cazuri, s-au nregistrat remisiuni ale semnelor clinice la 1 2
sptmni de la ncheierea tratamentului, asociate cu reapariia oochisturilor n
fecale. Remisiile nu au mai aprut dup un al 2-lea tratament cu anticoccidiene.
Din studiul efectuat, rezid importana diagnosticului infeciilor eimeriene,
inclusiv cu Isospora la carnivore, n special la tineret, care este n mod particular
sensibil i expus la multiple infecii. Existena bolii n cresctorii duce la creterea

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presiunii infecioase (alturi de ali ageni patogeni), cu riscul apariiei de enzootii


de boli cu manifestri digestive n perioada nrcrii sau cu ocazia unui stres
(vnzare, transport).
n mod particular, n cresctorii este necesar un diagnostic etiologic corect
i instituirea unui tratament susinut cu repetri periodice pentru reducerea
contaminrii mediului i evitarea exprimrilor clinice.
Un diagnostic etiologic corect i rapid la animalele cu diaree scurteaz
considerabil evoluia clinic a bolii. Fiind uor de efectuat, examenul coproscopic
se impune ori de cate ori avem de-a face cu evoluia unui episod de diaree la
animale tinere, mai ales n perioada de nrcare.

BIBLIOGRAFIE

1. Andre H., Pierson P., Polak B., 2001 - La coccidiose a Isospora spp. chez le chien.
Le Point Veterinaire, No. 219: 32 - 37
2. Bourdoiseau G., 2003 - Comment realiser une coproscopie lors de diarrhee aigue
chez le chien et le chat, Le Nouveau Praticien Veterinaire, 14, 3: 25 26
3. Charles V., Trayser DVM, MS, Kenneth S. Todd.,Jr., PHD, 1978 - Life Cycle of
Isospora burrowsi spp. (Protozoa: Eimeriidae) from the Dog Canis familiaris,
American Journal of Veterinary Research, vol. 39. no. 1: 95 - 98
4. Conboy G., 1998 - Canine coccidiosis, Can Vet J, Vol 39,: 443 - 444
5. Dubey P.J., 1978 - Pathogenicity of Isospora ohioensis Infecion n Dogs, Jurnal of
the American Veterinary Association, vol.173, no 2: 192 197
6. Dubey J.P., Streitel R.H., 1976 - Isospora felis and I. rivolta infections n cats
induced by mouse tissue or oocysts, Br. vet. J., 132, 649 651
7. Dubey J.P.. Weisbrode E. Steven, William A. Pogers, 1978 - Canine Coccidiosis
Attributed to an Isospora ohioensis -like organism: A Case Report, Jurnal of the
American Veterinary Medical Association, vol.173, no 2: 185-192
8. Dulceanu N., 1980 - Coccidioze i alte sporozooze la animale, Ed. Ceres, Bucureti
9. Dulceanu N., Cristina Terinte, Mitrea I.L., Carmen Palcovnicu, 2002 - Dicionar
enciclopedic de parazitologie, Ed. Academiei Romne, Bucureti
10. Dunbar M., Foreyt J., 1985 - Prevention of coccidiosis n domestic dogs and captive
coyotes (Canis latrans) with sulfadimethoxine-ormetroprim combination, Am. J.
Vet. Res., vol. 46, 9: 1899 -1902
11. Lloyd S., Smith J., 2001 - Activity of toltrazuril and diclazuril against Isospora
species n kittens and puppies, The Veterinary Record, 16: 509 - 511
12. Mitrea I.L., 2002 - Boli parazitare la animale, Ed. Ceres, Bucureti, 2002.
13. OBrien C.R., Pope S.E., Malik R., 2002 - Vomiting, diarrhoea and inappetence n a
Young cat with hypoproteinaemia, Aust. Vet. J., vol.80, 9:544-551
14. Omata Y., Oikawa H., Kanda M., Mikazuki K., Florencia G. Claveria, Cecilia
Dilorenzo, Takehara T., Saito A., Suzuki N., 1991 - Humoral Immune
Response to Isospora felis and Toxoplasma gondii n Cats Experimentally
Inoculated with Isospora felis, J. Vet. Med. Sci. 53(6): 1071-1073
15. Pierson P., 2003 - Conduire a tenir face a un episode de diarrhee aigue en elevage
canin, Le Nouveau Praticien Veterinaire, 14: 17-23
16. Pierson P., 2003 - Observaion clinique dune diarrhee aigue liee a une infection
parasitaire dans un elevage canin, Le Nouveau Praticien Veterinaire, 14: 33-35
17. Valkounova J., 1982 - Parasitological investigation of childrens sand boxes and dog
faeces from public areas n old housing districts of Prague, Folia Parasitologica
(Praha) 29:25-32

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INCIDENA HIDATIDOZEI N SUDUL I VESTUL


ROMNIEI N PERIOADA 1999 - 2004
THE INCIDENCE OF CYSTIC ECHINOCOCCOSIS IN WESTERN
AND SOUTHERN ROMANIA BETWEEN 1999 AND 2004

S. MORARIU, I. COSOROAB,
Gh. DRBU, I. OPRESCU, I. IACOBICIU,
Florica MORARIU, Narcisa RADBEA

Cystic Echinococcosis (CE) is one of the most important parasitic


zoonosis. In many parts of the world CE has an increased incidence. This
study reveals the incidence of CE in three districts of western and southern
part of Romania. Considering all slaughtered animals, the highest
incidence was recorded in Gorj district (7.60%), followed by Timi district
(4.20%) and Cara-Severin district (1.66%). Bovines were the most infested
species, with a highest incidence in Gorj district (48.38%), followed
decreasingly by Timi district (24.04%) and Cara-Severin district
(11.75%). CE had a highest incidence in pigs in Timi district (5.02%). In
sheep, CE prevailed in Gorj district (10.90%).

Key words: cystic echinococcosis, incidence, Timi, Cara-Severin, Gorj districts.

n Romnia, incidena hidatidozei este, n general, ridicat, dar diferit n


funcie de specia gazd i dup regiune. n urma nregistrrilor efectuate n
abatoare s-au raportat infestaii ce au variat ntre 10 i 65%. Dac s-ar lua, ns, n
calcul i prezena hidatidelor la animalele moarte sau sacrificate n afara
abatoarelor, cifrele ar fi mult mai mari.
n Europa, infestaia hidatic variaz de la o ar la alta. Astfel, n Bulgaria,
rata infestaiei a fost de 22,55% la ovine, 12,30% la bovine i 2,88% la suine (8),
n Iugoslavia a fost de 58,63% la ovine, 11,70% la bovine i 35,37% la suine (2),
iar n Italia, Lorenzini i Ruggieri (4) identific 0,7% bovine, 5,4% ovine i 0,4%
suine infestate.
Avnd n vedere aceste date i lipsa unor studii privind situaia
epizootologic a hidatidozei n vestul Romniei, s-a considerat oportun
efectuarea unei anchete epidemiologice n patru judee din zona de vest i sud-
vest a rii.

MATERIALE I METODE

Determinarea incidenei infestaiei cu chisturi hidatice la bovinele, ovinele i


suinele din partea de vest a Romniei s-a fcut pe baza datelor obinute de la
Direciile Sanitare Veterinare ale judeelor Timi, Cara-Severin i Gorj.
Rezultatele obinute sunt prezentate n tabelele 1, 2 i 3 i se refer la perioada
1999 - 2004.

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Tabelul 1.
Incidena infestaiei hidatice la animalele sacrificate n judeul Timi,
n perioada 1999 2004

Total animale Ovine Bovine Suine


examinate

examinate

examinate

examinate
Pozitive

Pozitive

Pozitive

Pozitive
Anul
(%)

(%)

(%)

(%)
47.962 87 2.430 45.445
1999 645.620 1.313 11.638 632.669
(7,42) (6,62) (20,87) (7,18)
11.076 141 1.857 29.078
2000 264.686 2.597 7.730 254.359
(4,18) (5,42) (24,02) (11,43)
7.033 112 721 6.200
2001 243.488 2.031 3.337 238.120
(2,88) (5,51) (21,60) (2,60)
2.720 97 803 1.820
2002 205.575 2.164 4.791 198.620
(1,32) (4,48) (16,76) (0,91)
6.317 117 419 5.781
2003 244.880 1.980 3.251 239.649
(2,57) (5,91) (12,88) (2,41)
2.401 102 382 1.917
2004 238.491 1.778 2.842 233.871
(1,01) (5,73) (13,44) (0,82)
77.509 656 6.612 90.241
TOTAL 1.842.740 11.863 33.589 1.797.288
(4,20) (5,53) (19,68) (5,02)

Tabelul 2
Incidena infestaiei hidatice la animalele sacrificate n judeul
Cara - Severin, n perioada 1999 2004.

Total animale Ovine Bovine Suine


examinate

examinate

examinate

examinate
Pozitive

Pozitive

Pozitive

Pozitive

Anul
(%)

(%)

(%)

(%)

1.348 450 531 367


1999 149.928 4.739 5.126 140.063
(0,89) (9,49) (10,35) (0,26)
2.068 347 526 1.195
2000 83.081 10.882 4.791 67.408
(2,48) (3,18) (10,97) (1,77)
1.041 249 424 368
2001 55.035 8.636 3.607 42.792
(1,89) (2,88) (11,75) (0,85)
239 98 49 92
2002 20.125 1.441 1.268 17.416
(1,18) (6,80) (3,86) (0,52)
710 375 142 193
2003 25.522 5.649 1.773 18.100
(2,78) (6,63) (8,01) (1,06)
545 209 161 175
2004 23.004 2.842 1.804 18.358
(2,36) (7,35) (8,92) (0,95)
5.951 1.728 1.833 2.390
TOTAL 357.417 34.189 18.369 304.137
(1,66) (5,05) (9,97) (0,78)

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Tabelul 3.
Incidena infestaiei hidatice la animalele sacrificate n judeul Gorj,
n perioada 1999 2004

Total animale Ovine Bovine Suine


examinate

examinate

examinate

examinate
Pozitive

Pozitive

Pozitive

Pozitive
Anul
(%)

(%)

(%)

(%)
1.888 108 839 941
1999 14.679 1.026 2.299 11.354
(12,86) (10,52) (36,49) (8,28)
1.883 384 778 721
2000 9.836 2.307 1.608 5.921
(19,14) (16,64) (48,38) (12,17)
483 259 82 142
2001 7. 252 1.872 653 4.727
(6,66) (13,83) (12,55) (3,00)
295 71 97 127
2002 6.865 958 572 5.335
(4,29) (7,41) (16,95) (2,38)
405 83 181 141
2003 11.750 1.429 870 9.451
(3,44) (5,80) (20,80) (1,49)
381 135 79 167
2004 19.746 1.943 609 17.194
(1,92) (6,94) (12,97) (0,97)
5.335 1.040 2.056 2.239
TOTAL 70.128 9.535 6.611 53.982
(7,60) (10,90) (31,09) (4,14)

REZULTATE I DISCUII

Ancheta epidemiologic realizat n judeul Timi relev o inciden


contrastant a hidatidozei la speciile de animale luate n studiu (tabelul 1 i figura
1). Astfel, la ovine i suine, valorile incidenei bolii sunt apropiate: 5,53%,
respectiv, 5,02%. n schimb, la bovine, aproape o cincime din animalele
sacrificate a fost parazitat (19,68%).
La ovine, cea mai mare inciden a fost constatat n anul 1999 (6,62%), iar
cea mai mic, n anul 2002 (4,48%). Cele mai multe bovine parazitate au fost
nregistrate n anul 2000 (24,02%), iar cele mai puine, n anul 2003 (12,88%).
n anul 2000 au fost consemnate statistic cele mai multe suine parazitate
(11,43%), iar n anul 2004, cele mai puine animale afectate (0,82%) din aceast
specie.
Surprinde totui procentul mare de suine afectate, avnd n vedere faptul c
pn n anul 1999 ponderea a fost deinut de suinele din Holdingul COMTIM,
unde msurile de igien erau mult mai atent aplicate i respectate. Pe msura
afluirii grsunilor din gospodriile populaiei, procentul animalelor infestate a
crescut, astfel c n anul 2000 a atins apogeul (11,43% animale parazitate).

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1999 2000 2001 2002 2003 2004

0,82
2,41
Suine 0,91
2,6
11,43
7,18
13,44
12,88
Bovine 16,76
21,06
24,02
20,87
5,73
5,91
Ovine 4,48
5,51
5,42
6,62

Fig. 1. Incidena hidatidozei (%) la animalele sacrificate n judeul Timi (A)


n perioada 1999 2004.

Contrastant este i numrul mare de bovine infestate. Peak-ul a fost


semnalat n anul 2000 (24,02%), dup care urmeaz o descretere semnificativ,
pn la 12,88%, n anul 2003.
Pentru judeul Cara-Severin, situaia este oarecum asemntoare cu aceea
din judeul Timi (tabelul 2 i figura 2). Ovinele au fost parazitate ntr-o proporie
apropiat de cea din judeul Timi (5,05%), iar la suine incidena este cea mai
mic (0,78%). i n judeul Cara-Severin, bovinele au fost mai intens parazitate
(9,97%), comparativ cu celelalte specii, ns incidena a fost mai mic dect n
celelalte judee (19,68% pentru Timi i 31,09% pentru Gorj).
La ovine, incidena maxim a fost consemnat n anul 1999 (9,49%), dup
care urmeaz o scdere treptat a cazurilor de hidatidoz, astfel c n anul 2001 a
fost nregistrat incidena minim (2,88%). n anii urmtori infestaia s-a situat la
un nivel mai ridicat, dar relativ constant, cu mici fluctuaii n jurul valorii de 7%.
La bovine, cele mai numeroase cazuri au fost semnalate n anul 2001
(11,75%), iar cele mai puine, n anul 2002 (3,86%), dup care valorile au revenit
la valori aproximativ egale n anii urmtori (8,01% i 8,92%). Aceast scdere
brusc n anul 2002, urmat de revenirea la valorile apropiate de cele anterioare,
rmne inexplicabil.
Pentru suine, rata infestaiei se menine sczut, cu un maxim de 1,77% n
anul 2000, urmat de o descretere treptat. Minimul infestaiei a fost nregistrat
n anul 1999, cu 0,26% animale parazitate. De altfel, suinele sacrificate n judeul
Cara-Severin au prezentat cea mai sczut rat a infestaiei dintre animalele
examinate n cele patru judee.

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0,95
1,06 1999 2000 2001 2002 2003 2004
0,52
Suine 0,85
1,77
0,26
8,92
8,01
3,86
Bovine 11,75
10,97
10,35
7,35
6,63

Ovine 6,8
2,88
3,18
9,49

Fig. 2. Incidena hidatidozei (%) la animalele sacrificate n judeul Cara-Severin (B)


n perioada 1999 2004.

n judeul Gorj, contrasteaz de la nceput numrul mic de animale


sacrificate cu coresponden n actele oficiale (70.128) cu procentele mari ale
infestaiei, cel puin pentru anii de debut ai anchetei.
Astfel, se poate constata cu uurin faptul c anul 2000 deine recordul
infestaiilor la toate cele trei categorii de animale investigate (16,64% la ovine,
48,38% la bovine, respectiv 12,17% la suine. Pare de necrezut c jumtate din
bovinele sacrificate sunt purttoare de hidatide, cel mai adesea sterile ns.

0,97 1999 2000 2001 2002 2003 2004


1,49
2,38
Suine 3
12,17
8,28

12,97
20,8
Bovine 16,95
12,55
48,38
36,49

6,94
5,8
7,41
Ovine 13,83
16,64
10,52

Fig. 3. Incidena hidatidozei (%) la animalele sacrificate n judeul Gorj (C)


n perioada 1999 2004.

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La ovine incidena minim a fost consemnat n anul 2003 (5,80%), la


bovine n anul 2001 (12,55%), iar la suine n anul 2004 (0,97%).
Se poate remarca i pentru acest jude c la suine numrul animalelor
infestate este mai mic. De fapt se situeaz pe locul doi ca procent minim de suine
infestate (4,14%), dup judeul Cara-Severin (0,78%), dar naintea judeului
Timi (5,02%).
Dac s-ar lua n considerare i animalele sacrificate n gospodriile
populaiei sau n diverse puncte de sacrificare mai mult sau mai puin clandestine,
n care controlul sanitar veterinar este sumar sau chiar lipsete, atunci procentul
animalelor infestate ar fi, probabil, mai mare.
Comparnd situaia existent n cele patru judee din vestul i sud-vestul
Romniei cu datele statistice din unele ri europene, mai ales din jurul
Mediteranei, putem spune c n ara noastr, infestaia se menine la o cot medie
fa de aceste ri. Astfel, n Bulgaria, rata infestaiei ovinelor a fost de 30,26% n
1996, iar la bovine, n funcie de zon, a variat ntre 9,2% i 25,9% (8); n Turcia,
30,6% la ovine i 25,9% la bovine, n perioada 2000-2001; n Grecia, 0,23-21% la
ovine i 0-53% la bovine, pe o perioad de 10 ani (1989 - 1998) (1); la nceputul
programului de control n Grecia, n 1984, prevalena infestaiei a fost de 82%
dintre bovine, 80% dintre ovine, 24% dintre capre i 5% dintre porci (6, 7); n
Italia, tot pe o perioad de 10 ani (1990 1999), la ovine a fost de 86,9% i
23,7% la bovine (4). n Spania, la nceputul programului de control, n 1986,
prevalena infestaiei la ovine a fost de 82,3%, iar n anul 2000 aceasta a ajuns la
doar 20% (3).

35
Ovine Bovine Suine Total anim ale
30

25

20

15

10

0
Tim i Cara Gorj

Fig. 4. Incidena cazurilor de hidatidoz (%) la animalele sacrificate


n perioada 1999 2004.

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Dac se analizeaz atent datele prezentate n cele patru tabele i n graficele


aferente fiecrui tabel, se poate observa cu uurin faptul c, n toate cele patru
judee, este nregistrat o scdere lent i progresiv a cazurilor de hidatidoz la
animalele de interes economic. Aceast scdere nu are la baz, ns, un suport
real. Ea se bazeaz, n mare parte, pe reducerea drastic a efectivelor de animale,
att n sistemul de stat, ct i n cel privat. De asemenea, efectuarea unor
tratamente antihelmintice generale cu substane benzimidazolice sau pro
benzimidazolice ar fi putut contribui ntr-o oarecare msur la reducerea
incidenei bolii. Factorii de mediu (temperatur, umiditate) par a fi avut i ei un
rol n limitarea transmiterii bolii. ncepnd cu anul 2002, verile au devenit din ce
n ce mai clduroase i secetoase. n astfel de condiii, supravieuirea oulor de
Echinococcus granulosus pe pune devine problematic, ceea ce duce la
diminuarea transmiterii hidatidozei.
Totui, meninerea infestaiei la cote relativ ridicate, poate fi explicat prin
faptul c toate cele trei specii menionate puneaz, n multe regiuni, mpreun,
iar cinii ciobneti i cei hoinari contribuie la macularea cvasipermanent a
punilor. Pentru zonele submontane i montane din cele trei judee nu trebuie
exclus nici intervenia canidelor slbatice, ca gazde definitive, n lanul
epidemiologic al parazitozei.

CONCLUZII

n toate cele trei judee luate n studiu: Timi Cara-Severin i Gorj,


incidena hidatidozei se menine la un nivel ridicat.
Cea mai mare rat a infestaiei hidatice este ntlnit n judeul Gorj:
7,60%.
Dintre cele trei specii de animale investigate, bovinele sunt cel mai intens
parazitate, cu un maxim n judeul Gorj (48,38%, fa de 24,02% n Timi i
11,75% n Cara-Severin). Aceasta nseamn un dublu de bovine infestate fa de
judeul Timi i de patru ori mai mult comparativ cu judeul Cara-Severin.
Diversele transformri n structura proprietii, tratamentele antihelmintice
cu substane benzimidazolice i presiunea de selecie exercitat de factorii de
mediu au contribuit la reducerea transmiterii hidatice.
Fluctuaiile anuale se pot explica i prin faptul c, n abatoare nu vin n
fiecare an animale din aceleai ferme sau localiti, iar aceast reducere a
incidenei nu poate fi apreciat dect aproximativ.

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BIBLIOGRAFIE

1. Altinta, N., 2003 Past to present: echinococcosis in Turkey. Acta Tropica, 85, 105
112.
2. Damnjanovic, D.; Kulisic, Z.; Pavlovic, I., 1995 - Prevalence hydatidosis in Valjeva
area. XVII International Congress of Hydatidology, 6 10 November, Limassol,
Cyprus.
3. Jimenez, S.; Perez, A.; Gil, H.; Schantz, P.M.; Ramalle, E. ; Juste, R.A., 2002
Progress in control of cystic echinococcosis in La Roja, Spain: decline in
infection prevalences in human and animal hosts and economic costs and
benefits. Acta Tropica, 83, 213 221.
4. Lorenzini, R.; Ruggieri, A., 1987 - Distribution of echinococcosis - hydatidosis in
Italy. J. Helminthol., 61, 3, 261 267.
5. Morariu, S.; Cosoroab, I.; Drbu, Gh.; Florica Morariu; Oprescu, I., 2005
The incidence of hydatid disease in western part of Romania. Lucr. t. Med. Vet.
Timioara, XXXVIII, 598 - 603.
6. Seimenis, A., 2003 Overview of the epidemiological situation on echinococcosis in
the Mediterranean region. Acta Tropica, 85, 191 195.
7. Sotiraki, S.; Himonas, C.; Korkoliakou, P., 2003 Hydatidosis echinococcosis in
Greece. Acta Tropica, 85, 197 201.
8. Zheliaskov, P.; Tanchev, T.; Toncheva, A., 1995 Epizootological investigation an
Echinococcosis Hydatidosis among dogs and animals intermidal hosts in
some parts in Bulgaria. XVII International Congress of Hydatidology, Limassol,
Cyprus.

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THE NEMATODE OF RED FOXES (VULPES VULPES L.)


HUNTED IN BELGRADE AREA
INFESTAIA CU NEMATODE LA VULPILE (VULPES VULPES L.)
VNATE N ZONA BELGRADULUI

I. PAVLOVI1, Z. KULI2, S.STANOJEVI1


1
Scientific Veterinary Institute of Serbia, Belgrade, Yugoslavia
2
Institute of Parasitology, Faculty of Veterinary Medicine, Belgrade

Large number of foxes, known to carry various parasites, lives in the


vicinity of Belgrade. Therefore we conducted a postmortem parasitological
(patho-anatomic) examination of a total of 719 adult and 156 fox puppies
hunted in the Belgrade area. Ten species of nematodes were found:
Toxocara canis, Toxocara mystax, Toxascaris leonina, Ancylostoma
caninum, Uncinaria stenocephala, Spirocerca lupi, Capillaria plica,
Rictularia affinis, Gonglyonema pulchrum, Trichinella spiralis and
Trichuris vulpis. Comparing these results with the results of examination of
stray dogs we concluded that many nematoda species were found in both
populations. This is of great epizootological and epidemiological
significance in terms of expansion of helminthoses, because of possible
urban environment contamination and subsequent human infection,
particularly with Ascaridae and Ancylostomidae species.

Key words: Fox, nematodes, dog, epidemiology

INTRODUCTION

In most European countries and as in many other parts of the world,


research surveys have been carried out to study of helminthes fauna of the fox
(Petrov & Potehina 1953; Irgaov & Sadikov,1966; Lubke,1972; Dent,1974;
Williams,1976; Hinaidy,1976; Kozlov,1977; Jancev&Ridjakov,1977; Edwards et
al.,1979; Nickel et. al.,1980; Borgstede,1984; Petavy et al.,1985; Ballek et
al.,1992; James & Walters,1992; Willingham et al.,1996; Papadopulos et al.,1997;
Shimalov & Shimalov,2003).
In Serbia, in Belgrade area, some information regarding the helminthes of
foxes has been given by Lozani, 1965; Bokovi & Valter,1979; Pavlovi.1994.
and Pavlovi et al.,1997. Those results represented that foxes in Belgrade area
were carriers of many species of nematode, including same species of importance
for human health.

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MATERIAL AND METHODS

In the period 1994 - 2000 a total of 875 red foxes (Vulpes vulpes L.) (719
adult and 156 puppies) were examined in the laboratory of Veterinary institute in
Belgrade. Foxes were hunted at spread area of Belgrade.
After necropsy, we have examined trachea, lung, heart, complete
gastrointestinal tract, liver, kidney and urinary bladder. The stomach, intestine and
the other organs were slit opened and visible helminthes removed. Afther that
(especially to intestine) the contents washing out. The contents and washing were
scened over a gauze sieve, mesh aparature 150 mm, under jet water and the
retained material examined, small quantities at a time, in a large whistle enamel
tray. Found parasites were fixing in 10% formalin, were mounted in lactophenol
for identification, and mounted in Canada balsam. Determination we have done
by keys given by Watson (1963) and Soulsby (1977).

RESULTS AND DISCUSSION

During examination we recorded parasitic infection in 91,08% (797) foxes,


caused by 5 trematoda species, 11 cestoda and 11 species of nematode. Mixed
infection was more common then single ones. Poliparasitic infections were found
at 706 (80.68%) foxes. Infections with nematoda were found at 703 (80.34 %)
adult foxes. Puppy foxes were infected with only one nematoda species.
Toxocara canis was found at 62 (39,74%).
Table 1
Nematode infections of red foxes in spread area of Belgrade1988-2003

Number Prevalence Number of helminthes


Nematode species
infected (%) (min max)
Toxocara canis 383 43,78 2-29
Uncinaria stenocephala 245 28,00 6-112
Trichuris vulpis 184 21,57 4-23
Ancylostoma caninum 166 18,91 2-18
Toxocara mystax 94 10,74 3-8
Trichinella spiralis 54 6,17 -
Capillaria plica 47 5,37 2-71
Rictularia affinis 45 5,14 0-9
Toxascaris leonina 29 3,31 3-15
Spirocerca lupi 9 1,05 2-3
Gonglyonema pulchrum 1 0.01 11

The results we obtained in our examination have shown that a considerable


number of foxes carriers of number of nematoda species. The most prevalent
among the species was Toxocara canis which was observed in 43,78 % of adult
and 39,74 % puppy foxes. Followed by Uncinaria stenocephala (28,00%),
Trichuris vulpis (21.57%), Ancylostoma caninum (18.91%), Toxocara mystax
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(10,74%), Capillaria plica (5,37%), Trichinella spiralis (6,17%), Toxascaris


leonina (3,31%) Rictularia affinis (5,14%), Spirocerca lupi (1,05%) and
Gonglyonema pulchrum at 0,01% of the foxes examined (table 1).

CONCLUSION

Foxes inhabited in Belgrade area are carriers for many nematoda species,
especially Ascaridae and Ancylostomidae, which caused human larval syndrom.

REFERENCES

1. Balek, D., Takla, M., Ising-Voler, S. & Stoye, M., 1992 Zur Helminthenfauna des
Rotfushes (Vulpes vulpes, Linne, 1758) in Nordhessen und Ostewestfalen, Teil
2: Nematoden. Deutsche Tierarztliche Wochenscrift 99, 435-437.
2. Bjotvedt, G., Tomikiewitcz, S.M.Jr. & Goliqhtly, R.T.Jr., 1980 - A description of
helminths of kit fox (Vulpes macrotis). Veterinary Medicine and Small Animal
Clinical 75 (5), 661-884.
3. Borgsteede, F.H.M., 1984 - Helminths parasites of wild foxes (Vulpes vulpes L.) in
the Nederlands. Zentralblat fur Parasitekude 70, 281-285.
4. Bokovi, V.& Valter, D., 1979 - Parazitska fauna lisica i jazavaca sa podruja
Beograda. Veterinarski glasnik 33 (12), 1023-1025.
5. Coman, B.J., 1973 - Helminth parastes of the red fox (Vulpes vulpes) in Victoria.
Australian Veterinary Journal 49, 378-384
6. Deblock, S., Petavy, A.F. & Gilot, B., 1988 - Helminthes intestinaux du renard
commun (Vulpes vulpes L.) dans la Masif Central (France). Canadian Journal of
Zoology 66, 1562-1569.
7. Diebble, E.D., Font, W.F. & Wittrock, D.D., 1983 - Helminths of red fox, Vulpes
vulpes L. in west central Wisconsin. Journal of Parasitology 69 (6), 213-214.
8. Hackett, F. & Walters, T.M., 1980 - Helminths of the red fox in Mid-Wales,
Veterinary Parasitology 7, 181-184.
9. Hinaidy, H.K., 1976 - Ein Weiterer Beitrag zur Parasitenfauna des Rotfuches Vulpes
vulpes (l) in Osrterreich. Zentralblat fur Veterinaremedizine B, 23, 66-73.
10. Irgaov, I.H. & Sadikov, V.M., 1966 - O gelmintofaune lisic Uzbekistana. Materiali k
naunoj konferensii vsesojouznogo obsestva gelmintologov, Tom III, 125-126.
11. Jancev, J. & Ridjakov, N., 1977 - Helminth fauna of the fox (Vulpes vulpes rucigera
Becstein) in Nortwestern Bulgaria. Helminthologia Bulgaria 4, 73-96.
12. Koanov, E.K., 1972 - Gelminti dikih mlekopitajunih Uzbekistana, Materiali
Akademii Nauk Uzbekskoi SSR, Takent, 72-73.
13. Kovaluk, E.S., 1974 - Gelminti mlekopitajunih v Tjomenskom priirtisje,
mlekopitajunih analiz, Izdatelstvo Ministarstvo hojzovstva SSSR, Moskva.
14. Kozlov, D.P., 1977 - Opredeljitel gelmintov hinih mlekopitajunih SSSR. zdatelstvo
Akademii Nauk SSSR, Moskva.
15. Loos-Frank, B. & Zeyhle, E., 1982 - The intestinal helminths of the red foxes and
some other carnivores in south germany. Zentralblat fur Parasitekude 67, 99-
113.
16. Lozani, B., 1961 - Prilog poznavcanju helmintoza lisica, I nematode. Vetrinarski
glasnik 15 (3), 207-209.

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17. Lozani, B., 1965 - Contribution to know of helminthose of foxes (Vulpes vulpes)
from hunting area of spread area of Belgrade. Doctoral thesis, Faculty of
Veterinary Medicine, University in Belgrade.
18. Nickel, S., Hiepe, T., Hansel, U. & Jurke, E., 1980 - Contribution to the parasitic
fauna of the GDR, Investigation of the prevalence of the helminths in red fox
(Vulpes vulpes L.). Angewandt Parasitologie 21 (2), 94-100.
19. Papadopoulos, H., Himonas, C., Papazahriadou, M. & Asntoniado -Satiroadou K.,
1991 - Helinthews of foxes and other wild carnivores from rural area of Greece.
Journal of Helminthology 71, 227-231.
20. Pavlovi, I., 1994 - Helmnthoses of red foxes (Vulpes vulpes L.) hunting at Belgrade
area in period 1988-1992., Doctoral thesis. Faculty of Veterinary Medicine,
University in Belgrade.
21. Pavlovi, I., Kulii, Z. & Milutinovi, M., 1997 - The role of foxes (Vulpes vulpes
L.) in the epizootiology and epidemiology of nematoda parasitic zoonoses. Acta
Veterinaria 47 (2-3), 177-182
22. Petavy, A.F.& Deblock, S., 1980 - Helminthes of the common fox (Vulpes vulpes)
from Masif Central (France). Annales de Parasiotologie Humane et Comparee 55
(4), 379-391.
23. Petavy, A.F., Deblock, S., Cantat, F. & Gilot, B., 1985 - Le renard et ses
helminthes et France. Revue d Ecologie la Terre et la Vie 40, 231-237.
24. Petrov, A.M. & Potehina, L.F., 1953 - K gelmintofaune hinih mlekopitajunih
Tadikistana. Trudi Vsesojouznoga instituta gelmintologov imena Skrajbin 5,
167-172.
25. Ryan, G.E., 1976 - Helminth parasites of the fox (Vulpes vulpes) in West South
Wales, Australian Veterinary Journal 52, 126-130.
26. Richards, D.T., Harris, S. & Lewis, J.W., 1995 - Epoidemiogical studies on
intestinal helminth parasites of rural and urban red foxes (Vulpes vulpes) in the
United Kingdom. Veterinary Parasitology 59, 39-51.
27. Scoffel, I., Schein, E., Wittstadt, U. & Hentsce, J., 1991 - Zur parasitenfauna des
rotefuschs in Berlin (West). Berliner Tierarztliche Wochenschrift 104, 153-157.
28. Soulsby, E.J.L., 1977 - Helminths, arthropods and protozoa of domesticated
animals, Baille, Tindall and Cassell Co, London.
29. Williams, B.M., 1976 - The intestinal parasites of the red fox in South Wales. British
Veterinary Journal, 132 (3) 309-312.
30. Willingham, A.L., Ockens, N.W., Kapel, C.M. & Monrad, J., 1996 - A
helminthological survey of wild red foxes (Vulpes vulpes) from the metropolitan
area of Copenhagen. Journal of Helminthology 70, 259-263.
31. Wessbecher von H., Dalchow W. & Stoye, M., 1994 - Zur helmintenfauna des
Rotfuisches (Vulpes vulpes Linne 1758) in Regierungsbezirk Karlshrue, Teil 2:
Nematoden, Deutsche Tierarztlishe wochenschrift 101, 362-364

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EVOLUTION OF CANINE DEMODICOSIS IN DOGS


WITHOUT ACARICID TREATMENT
Narcisa RADBEA*
U.S..M.V. Timioara

We study 14 dogs diagnosticated with canine demodicosis. The dogs


were diagnosed with LD (9 cases), GD (3 cases) and PD (2 cases). The
therapy consisted in: animal preparation (deparasitation for helmintic
parasites, general antibiotherapy for PD, balanced nutrition, optimal
hospitalising conditions); daily wadding over the whole body with an acid
solution (apple vinegar+honey 100 ml+ sunflower oil 100 ml+ 800 ml
water); no topical or systemic acaricide was administered. Changing of the
microclimate and of the life conditions, ageing, shifting of the skin pH by
daily wadding over with an acid solution (apple. vinegar with
honey+oil+water) are possible causes of healing with LD and of the
circumscribing of the lesions in a dog with GD, in the absence of the
acaricide treatment. In the case of the 2 dogs diagnosed with PD, systemic
antibiotherapy associated with local application of the acid solution (apple
vinegar) determined the exacerbation of the purrulent lesions.

Canine demodicosis treatment has long passed the era of prescribing cedar
or Chaulmoogra oils . At present time, antidemodecic therapy involves a complex
of factors and attitudes: analysis of exach case, exmination of individual factors,
diagnosis of the clinical evolutive form (localised, generalised demodicosis or
piodemodicosis), evaluation of the general clinical state of the animal, the
individualisation of the therapeutic protocols, choosing of the acaricide by the
reactions of the animal, the extension and gravity of the lesions, the cost and the
commodity in administration. Localised demodicosis (LD) is considered an easy
form that heals spontaneously in 80 90% of cases (1, 2), whereas, in the general
non infected form (GD), spontan healings are reported to 20 50% of cases,
especially in dogs aged from 1 to 2 years (6). Piodemodicosis (PD) is
unanymously considered a severe evolution form, difficult to treat (1, 2, 3, 4, 5,
6).
Considering these references, this pioneering study followed the evolution
of canine demodicosis without acaricide treatment.

MATERIALS AND METHOD

Due to the fact that initally we havent had the possibility of follow up for
spontan healings in owned dogs, he have started the experiment with 14
unowned dogs. The dogs were diagnosed with LD (9 cases), GD (3 cases) and

*
E-mail: narcisa.radbea@fmvt.ro

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PD (2 cases). The therapy consisted in: animal preparation (deparasitation for


helmintic parasites, general antibiotherapy for PD, balanced nutrition, optimal
hospitalising conditions); daily wadding over the whole body with an acid solution
(apple vinegar+honey 100 ml+ sunflower oil 100 ml+ 800 ml water); no topical or
systemic acaricide was administered.
All the dogs were registered at the Clinic of Parasitic Diseases of the
University for 6 months. After release, they were clinically and parasitologically
monitored.

RESULTS AND DISCUSSION

At 4 months from the beginning of the treatment I have noticed interesting


results: 2 dogs diagnosed with LD presented the first signs of clinical healing
(apparition of hair in initially alopecic areas); in one dog diagnosed with GD I
have noticed the localisation of initially generalised lesions; in the 2 dogs
diagnosed with PD the clinical signs of healing did not appear, moreover, I have
noticed violent reactions to the application of the acid solution, that behaviour
determined me to stop administering the acid solution and to continue antibiotic
treatment.
At the end of the treatment (after 6 months), the situation was unchanged: 2
dosg with LD clinically healed with no tendance to recurrence, one dog with GD
in which the initially generalised lesions have healed, 2 dogs with PD in which
the lesions were drying and 9 dogs (7 with LD and 2 with GD in which the initial
lesions agravated).
Every dog which, at the end of the initial treatment, received acaricide
(Amitraz, Ivermectine) healed clinically and parasitologically in the next 6
months.
The problem of spontan healing in canine demodicosis is still subject to
controversy. Havrileck et al. (1990) claims that there is no difference between the
evolution of LD with or without treatment, and the time necessary for remissions
and healings is imprevisible. Some authors (Scott et al., 1995) do not find the
tratment with acaricide substances in LD justified, because, in their opinion, this
would favour the development of a population of antiparasitic resistant acariens.
These affirmatios do not answer the main problem of initial parasite proliferation:
why some of the dogs with the dry forms heal spontaneously, while others do not
react to acaricides; from the percent of the healed dogs, how much do the
efficiency of the treatment represent and whch percentage is due to spontan
healing?
We reinforce these opinions with the claim that, from the practical point of
view, it is difficult to foresee the healing or the extending of the disease.
Moreover, the owners are not content with the situation when the physician waits
for spontaneous healing, without administating no medicine to the ill dog. The
same attitude is found at the owner if the physician administered only an adjuvant

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therapy (antihelmintics, antiseptics, antiseborrheics, immunostimulants) in


expectative to spontaneous healings.
In the present study, the apparition of clinical healing in two dogs with LD,
the transformation of generalised lesions present to a dog in circumscrise lesions
and the persistance of the lesions to the dogs with PD support multiple
explanations. The healings in the dogs with the dry form can owe, on one side, to
the changing of the microclimate, of the anterior life conditions (deficitary diet,
crowding, precarious hygiene conditions, intense parasitism) with new life
conditions (balanced diet, rich in vitamins and minerals, internal and external
deparasitations, and individual hosting spaces with the possibility of outdoor
exercise).
The alkalinisation of the skin pH, modifications to the composition of the
lipidic layer, hydrolising oh the sterols and the lipidic esthers, the main
components of the sebum, the increase of the concentration of fatty acids at the
skin surface are reactions (signalled by some authors) that characterise the
demodecic skin (1, 4).
On the other hand, the healing of the dry lesions may be explained also by
the aging of the dogs (from 8 9 months at the beginning of the treatment to 16
18 months at the end). In both situations there is an increase of the canines
immunocompetence against the parasite and its effects.
To all these may be added the changes to the skin microenvironment by
daily wadding over the body with acid solution (apple vinegar) with the purpose
of shifting the skin pH to acid, as it becomes alkaline in the presence of the
acarian Demodex Canis.
In the case of the dogs diagnosed with PD (2 dogs), there is some
confirmations of early data that claim that adjuvant treatment (antibiotics,
antiseborrheics, acid solutions) do not heal the piodemodicosis (5, 6). The
iritation of the skin reduces, the purulent lesions dry, but the main cause of these
skin lesions, Demodex Canis, can not be eliminated only by antibacterial or
antiseborrheic treatment. Moreover, the association of antibiotics with the topic
acid solution has proven not convenient to the 2 dogs with PD; the primary
lesions produced by the acarian Demodex Canis, with the secondary bacterial
ones, have increased in contact with the acid solution and remitted only when I
ceased this therapy and continued with systemic antibiotherapy.
The presented reasons justify the small number of cases in which this last
experiment was carried out. In this situation, no conclusion can be drawn, but the
wise choise is to report the results and to leave place for interpretations.

CONCLUSIONS

Changing of the microclimate and of the life conditions, ageing, shifting of


the skin pH by daily wadding over with an acid solution (apple. vinegar with

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honey+oil+water) are possible causes of healing in 2 dogs with LD and of the


circumscribing of the lesions in a dog with GD, in the absence of the acaricide
treatment.
In the case of the 2 dogs diagnosed with PD, systemic antibiotherapy
associated with local application of the acid solution (apple vinegar) determined
the exacerbation of the purrulent lesions.

REFERENCES

1. Cadiergues, Marie-Christine, Franc, M., 1995 La dmodcie canine, Rev. Md.


Vt., 171, 6/7: 383.
2. Guagure, E., 1995 La dmodcie du chien adulte. A propos de 22 cas. Prat.
Md. Chir. Anim. Comp. 26: 411.
3. Hamann, F., Wedell, H., Bauer, J., 1997 Zur Demodikose des Hundes,
Kleintierpraxis, 42, 9: 745.
4. Havrileck, B., 1990 Contribution a ltude de la dmodcie canine, Laction Vt.,
11: 1150.
5. Kwochka, K.W., Kunkle, G.A., Foil, C.O., 1986 Canine demodicosis, Current
Vet. Therapy., IX, 9: 531.
6. Scott, D.W., Miller, W.H., Griffin, C.E., 1995 Small Animal Dermatology, 5th
edition., W.B. Saunders Co., Philadelphia.

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IMUNOHISTOCHEMICAL RESULTS IN CANINE


PIODEMODICOSIS
Narcisa RADBEA1*, O. MEDERLE2,
Gh. DRBU1, I. OPRESCU1,
S. MORARIU1, M. ILIE1, D. IZVERNARIU2
1
U.S.A.M.V. Timioara
2
U.M.F. Timioara

We study 20 dogs, 10 infected with Demodex canis mite


(piodemodicosis) and 10 control group. We performed
immunohistochemmically method using LSAB2 system with following
antibody: Langerhanh cells, Lymphocyte and citokeratine, visualization
with DAB. Immunohistochemical staining reveals an intense process of
neutrophils and eosinophils migration as well as a hiperplasia of
melanocytes and macrophages. In piodemodicosis, there is a certain
modification of the skin: the number of the keratinocytes and Langerhans
cells decreases and that of the B cells increases as a consequence of the
umoral immune response.

INTRODUCERE

The Immunohistochemical diagnosis reveals only immune response in skin


canine demodicosis. There for he must associated with histopathological and
cytopathological exams to complete diagnosis.

MATERIALS AND METHODS

Skin biopsies from each dog of the two series (one of 10 dogs infected with
Demodex Canis and one of 10 healthy dogs used as control) were harvested and
immunohistochemmically stained using the LSAB2 system. Langerhans cells and
lymphocytes were primary immunophenotypised by the S100 protein. Citokeratin
CK and pan citokeratin with DAB visualization were used to mark the presence
of keratinocytes and melanocytes of the basal layer. Fibroblasts and basal cells
were identified by vimentin and, respectively, CK 20.

RESULTS AND DISCUSSION

Immunohistochemical staining revealed interesting aspects concerning the


presence of immunocompetent cells (Langerhans cells, keratinocytes), and the
reaction of dermis and epidermis of those cells as a response to he foreign body:
Demodex Canis.

*
E-mail: narcisa.radbea@fmvt.ro

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The complication of demodicosis by bacterial infections generate an


immunohistochemical picture characterised by the amplification of celular
reactions: hiperplasia and migration of the Langerhans cells towards the basal
layer and the dermis, intensely stained keratinocytes, vascular proliferations
accompanied by the hypertrophy of the fibeoblasts and of the basal cells, presence
in high numbers of granulocytes, B type lymphocytes and melanocytes. This
acute cellular response is present in all the members of the series with non
signifiant differences between individuals..
In the control series, immunohistochemical staining revealed the presence
of immunocompetent cells (Langerhans cells), of keratinocytes, and also basal
cells and fibrocytes, without any modification of number and cellular shape
(Figure 1, 2, 3, 4).

Fig. 1. - Hyperplasia of keratinocytes


Fig. 2. - Keratinocytes hyperplasia.
immunopositive with pancitokeratina

Fig. 3. - Keratinocytes and Langerhans cel Fig. 4. - Langerhans cel migrations into
hipertrophia horny layer

Immunologic cellular reactions from skin lesions in demodicosis are


studied by a considerable number of authors (1, 2, 3, 4). T type lymphocytes,
keratinoctes and Langerhans cells are most involved in skin immune response:
Langerhans cells and keratinocytes present the antigen Demodex to T cells, they
respond by he inflammatory reactions induced into the folliculary epithelium. By

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interleukin synthesis, the Langerhans cells and the keratinocytes start an


avalanche of messages that lead to migration of immunocompetent and
inflammatory cells.
Most of the authors (3, 4) claim that, besides the dry form of
demodicosis, the intensity of cellular reactions gradually increases and becomes
strong in piodemodicosis. Other authors (1, 2) associate the evolution of
demodicosis, from the localised form to the generalized one and then to
piodemodicosis with an immunosupression, which yields a minimal cellular
reaction that becomes absent during the evolution of the disease.
The immunohistochemical images obtained in the present study show an
immune response of ummoral type, marked by an increased number of B cells, a
simultaneous decrease of T cells, keratinocytes and Langerhans cells number,
and an obvious intervention of granulocytes and melanocytes.
The immunocompetent cells response (reaction of Langerhans cells,
keratinocytes and T cells) as well as the umoral reaction by the B cells,
neutrophils, eosinophils and macrophages are normal reactions to the presence of
a foreign body - Demodex Canis and later the bacterial flora in the dogs skin.

CONCLUSIONS

In piodemodicosis, there is a certain modification of the


immunohistochemistry of the skin: the number of the keratinocytes and
Langerhans cells decreases and that of the B cells increases as a consequence of
the umoral immune response.
Immunohistochemical staining reveals an intense process of neutrophils
and eosinophils migration as well as a hiperplasia of melanocytes and
macrophages.

REFERENCES

1. Caswell J.L., 1995 - Establishment of Demodex canis on canine skin engrafted onto
severe combined immunodeficient / beige mice. In: Investigation of the
immunopathology of canine demodicosis, pp. 96107. DVSc Thesis, University
of Guelph, Guelph, ON, Canada.
2. Caswell J.L., Yager J.A., Ferrer L., Weir J.A.M., 1995 - Canine demodicosis: a re-
examination of the histologic lesions and description of the immunophenotype of
infiltrating cells. Vet Dermatol 6:9-19. 1995
3. Ferrer L., Fondevila D., Rabanal R., Ramis A., 1992 - Detection of T lymphocytes
in canine tissue embedded in paraffin wax by means of antibody to CD3 antigen.
J Comp Path-ol 106:311-314.
4. Nickoloff B.J., Turka L.A., 1994 - Immunological functions of non-professional
antigen-presenting cells: new insights from studies of T-cell interactions with
keratinocytes. Immunol Today 15:464-469, 1994

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DE LA MALARIOTERAPIE LA PARAZITICA (1927-2005):


PROGRESE N PARAZITOTERAPIE I COMBATERE
BIOLOGIC
FROM THE MALARIO-THERAPY TO PARAZITICA (1927-2005): PROGRESS IN THE
PARASITO-THERAPY AND THE BIOLOGICAL CONTROL

E. UTEU

The present project outstands the progress of the past 8 decades


regarding the use of parasites to mans benefit.
We have obtained results in parasitotherapy, consisting of parasite
samples with oncostatic, anti-parasitic and immunomodelling effects in the
treatment of other ailments, such as Crohn disease. Host self-deparasitation
occurs under certain conditions, as a consequence of some immunological
phenomena, of hypersensitivity and so on.
Biological treatment has developed by means of the use of
hyperparasites in fighting parasitosis and interspecific hybrids, reflected in
the control of ticks and hypodermosis.

n medicina popular, utilizarea paraziilor i substanelor parazitare, pentru


efectele lor benefice, se cunoate din vremuri ndeprtate. n terapia
anticoagulant i hipotensiv se aplicau pe corpul bolnavilor lipitori (Hirudo
medicinalis, parazit facultativ), cu aciune hematofag. Apoi din glandele salivare
ale acestora a fost extras hirudina (hemofilina) i utilizat n farmacologie (21).
ns un eveniment medical important, cu impact asupra oamenilor de
tiin, s-a produs n anul 1927, cnd profesorului J. Wagner-Jauregg din Viena i
s-a acordat premiul Nobel pentru descoperirea efectului curativ al hematozoarului
palustru (Plasmodium malariae) la pacieni cu demen paralitic, tabes (sifilis
nervos) (19). Inocularea bolnavilor cu P. malariae era urmat de un sindrom
febril, cu efecte treponemocide. Aceast aciune, recunoscut ca malarioterapie
(5, 13, 19), a reprezentat o nou direcionare a cercetrilor parazitologice, n
scopul valorificrii paraziilor n terapie sau alte ramuri, cu efecte benefice. La
trecerea a opt decenii de la aceast important descoperire, pn la apariia
primului volum - Parazitica, n 2005 (39) - despre folosirea paraziilor cu
aciuni benefice n activitatea uman au avut loc cercetri i rezultate tiinifice
notabile. Din multitudinea rezultatelor obinute n acest interval, n reputate centre
de cercetare tiinific, ne vom referi la cele de: 1) parazitoterapie, 2)
autodeparazitare i 3) combatere biologic.

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1. PARAZITOTERAPIA

S-a dezvoltat i diversificat treptat, echipe de cercettori recurgnd la


utilizarea unor specii de parazii sau extracte parazitare, pentru aciunile lor n
oncoterapie, terapie antiparazitar sau n afeciuni cu alt etiologie (22, 18, 19).
1.1. Oncoterapia. Exist parazii cu aciune cancerigen, ns exist i
specii a cror prezen n organismul gazdelor are influen antitumoral. Keller i
colab. (cit. 39) au observat la oareci i obolani cu boal canceroas, infestai cu
Nippostrongylus brasiliensis sau cu Schistosoma mansoni, c s-a produs involuie
tumoral. Spaldanova i colab. (31) au constatat c la loturi de oareci infestai
experimental cu chiti de Trichinella spiralis sau T. pseudospiralis, crora li s-a
administrat, la 7-25 zile p.i., suspensie de celule de melanom B16, s-a produs
treptat o regresie tumoral, comparativ cu loturi martor, la care dezvoltarea a fost
mai mare. T. pseudospiralis a avut efect antitumoral mai puternic.
n 1978, King (cit. 39) a experimentat extracte de mduv osoas i de
splin, obinute de la taurine infectate cu Babesia bigemina, Theileria mutans
sau/i Anaplasma marginale, pe oareci inoculai cu virus fibrosarcomatos.
Rezultatele obinute, comparativ cu oareci martori, injectai cu aceleai extracte
de la taurine sntoase, au demonstrat c extractele de la animalele infestate au
coninut factori cu aciune citotoxic, antitumoral. Efectul protectiv (antitumoral)
cel mai ridicat (81-91%) l-a avut extractul medular de la cazuri cu babesioz;
urmate de cele cu theilerioz (58-69%) i foarte redus - cele cu anaplasmoz.
Extractele splenice au fost cu aciune citotoxic moderat. Mecanismele n-au
putut fi elucidate.
Diveri cercettori au experimentat lizate de parazii privind efectele
anticanceroase. Miyahara i colab. (23) au obinut un lizat antigenic de
Toxoplasma (Toxoplasma lysate antigen) care a fost injectat la loturi de oareci cu
tumori cutanate (induse prin injectare de metilcolantren). Tratamentul s-a fcut n
dozaje diferite, repetabile la 7 zile. Dimensiunile tumorale cele mai mici, fa de
ale martorilor, s-au obinut la loturile injectate cu 100 mg TLA/oarece. Aceiai
autori (22) au demonstrat, experimental, c oareci sensibilizai iniial cu TLA
induc celulelor splenice inoculate altor animale efecte oncostatice. Autorii
recomand folosirea produsului n oncologie veterinar.
1.2. Terapia antiparazitar. Dezvoltarea paraziilor n organismul gazd,
simultan cu ali germeni, declaneaz fenomene extrem de diferite:
competivitate, potenri patogene, antagonisme .a. Procesele antagonice
demonstreaz c unii parazii sau extracte din acetia pot fi utilizai cu efecte
terapeutice. Astfel, s-a observat c Toxoplasma gondii inoculat la oareci
determin o inhibare a dezvoltrii infeciei cu Plasmodium berghei, de unde
cercetrile au urmrit valoarea antiparazitar a lizatelor de Toxoplasma (TLA), de
Babesia sau de Theileria.

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TLA a avut efect antimalaric la oareci (Omata i colab., cit. de 39).


Administrat la oareci cu babesioz experimental (B. rodhaini), unic sau asociat
cu limfokine de la oareci sensibilizai n prealabil cu Babesia sau Toxoplasma, a
avut efect preventiv antibabesian. Comparativ cu martori, supravieuirea
oarecilor a fost de 66%, iar la martori n-au rmas supravieuitori. Efectele
antibabesian, antimalaric i antitheilerian al TLA se datoresc unor factori multipli
celulari i moleculari (24, 29, 39).
1.3. Terapia altor boli. Observaii recente ale unor medici, cercettori din
SUA, au relevat c n rile neindustrializate populaiile sunt mai frecvent
infestate cu parazii intestinali, ns nu sufer de boala Crohn (ileit, enterocolit
cu etiologie incert). Populaiile din rile hiperindustrializate nu sunt infestate
frecvent, dar sunt mai afectate de boala Crohn (6, 20, 34). De aici premisa c
infestaiile intestinale pot avea aciune antiinflamatoare (42). Khan i colab.
(2002) au dovedit c nematode intestinale la oareci au avut efect ameliorant
asupra colitelor. La Universitatea din Jova, Summers i colab., n 2005 (34), au
experimentat pe pacieni cu boala Crohn, voluntari, Trichuris-terapia. Un numr
de 29 de bolnavi au fost infestai cu ou de Trichuris suis, per os, cte 2500 per
pacient, repetabil la 3 sptmni, timp de 24 sptmni. S-au monitorizat: tabloul
clinic i indicele de activare al bolii (CDAI). Post-terapeutic s-a observat o
descretere semnificativ a CDAI, la 79,3% (la 23 din 29 cazuri), fr a induce
fenomene adverse. Helminii administrai au redus procesele inflamatorii
intestinale, au diminuat rspunsul imun i ameliorat starea, influennd populaiile
biocenozei intestinale cu ameliorarea sntii bolnavilor (6, 34, 42).
Exist rezultate cu alte sindroame la animale imunosupresive, de
hipersensibilitate, n care s-au folosit extracte parazitare cu rezultate favorabile (1,
4, 12, 14, 17, 30).

2. AUTODEPARAZITAREA (SELF-CURE)

Este un fenomen medical foarte important prin semnificaie i din


perspectiv terapeutic. Uneori, att la om ct i la animale infestate, s-au
observat eliminri spontane de helmini intestinali, prin excremente (ascarizi,
strongili etc.) avnd drept cauze factori multipli. Se tie, n prezent, c intervin
trei grupe de factori: intraspecifici parazitali; intrinseci gazdei i exogeni
(ecologici). Unii autori consider c baza antagonismului este de natur imunitar,
o stare de hipersensibilitate sau paraimunitate (Eberman, 1961; Dorchies i colab.,
1994) sau ali factori.
Efectul distrugerii helminilor i al eliminrii spontane de ou degenerate a
fost semnalat, n premier mondial, de Gherman (7, 10), la pacieni cu
Hymenolepis nana, n faza de convalescen, ca manifestare a strii imunitare.
Apoi, Gherman i colab. (8, 9) au urmrit dinamica autovindecrilor spontane n
himenolepidoz la om i importana practic medical a prezenei oulor

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degradate n masa fecal. n acelai sens, Gherman (11) a descris mecanismele


patogenetice n giardioz (lamblioz) la om; precum i semnificaia patogen a
florei microbiene i faunei parazitare din ductele biliare i intestinale asupra
suelor de Giardia (11, 14).
Recent, Dorchies i Alzien (4) au semnalat autodeparazitarea ovinelor cu
trichostrongilidoze, ca efect al reinfestrilor cu Oestrus ovis. n estroz, se induce
o stare imunosupresiv la ovine, dar reinfestarea declaneaz hipersensibilitate
succedat de deparazitarea spontan de ali parazii, precum Trichostrongylidae
(Haemonchus, Ostertagia).
Remarcm faptul c instituirea malarioterapiei (n 1927) a fost urmat peste
timp de parazitoterapie i n cazul altor afeciuni: autodeparazitarea n
himenolepidoz la om 1960 (7, 9, 10), n trichostrongilidoze la ovine,
consecutiv reinfestaiei cu Oestrus ovis (4) i mai recent (2005) prin Trichuris
terapia la om n boala Crohn (6, 34).

3. COMBATEREA BIOLOGIC

Pe plan mondial s-au diversificat metodele de combatere biologic a


parazitozelor i duntorilor la om i animale. Dup decenii de utilizare a
chimiopreveniei, au aprut populaii de parazii rezistente, iar concomitent s-au
resimit efectele negative ale polurii chimice a ecosistemelor. Combaterea
biologic a aprut ca o alternativ necesar (36, 37). Din multitudinea mijloacelor
de combatere ne referim la aplicarea experimental a hiperparazitismului i la
metodele de sterilizare a unor specii parazite. n acest scop, cercetri
premergtoare de biochimie au relevat mijloacele de comunicare intraspecific
feromonale (15, 16, 18, 38, 40).
3.1. Hiperparazii n combaterea parazitozelor. Lucrrile iniiale au
urmrit o mai exact cunoatere a speciilor hiperparazite i a efectelor patogene
ale acestora asupra paraziilor (hipergazde). Apoi s-au experimentat metode de
cultivare n laborator i diseminarea lor n mediul exterior n zone populate cu
hipergazde. Sunt cercetri care au experimentat aciunea patogen a unor
protozoare, n special microsporidii, n controlul trematodozelor la peti (18, 26,
27, 28). La noi n ar s-au fcut experimentri cu microsporidia Unikarion,
hiperparazit al formelor larvare (sporociti, redii i cercari) de Diplostomum
spathaceum, care paraziteaz melcii, Limnaea spp. n controlul diplostomozei
(metacercarioza) petilor. Oel (1991; 1999) a obinut culturi de spori de
Unikarion care dispersate n bazine piscicole, populate cu melci infestai cu forme
larvare de D. spathaceum, au provocat mortalitate ridicat melcilor i, implicit,
ntreruperea ciclului evolutiv al parazitului. Ali autori au experimentat i ali
hiperparazii: Palmieri i colab. (28) au folosit Nosema strigeoidea n controlul
metacercariozei oculare la peti, cu rezultate favorabile.

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n controlul anaplasmozei, babesiozei i ectoparazitismului cu insecte


hematofage la bovine, n combaterea integrat Programul Poseidon s-au
cultivat n laborator himenoptere, Tachininaephagus, parazitoide ale larvelor i
pupelor de Stomoxys. Lansate n preajma fermelor de bovine, n decurs de 3 ani
(1995-1998), au produs scderea populaiilor de stomoxine i reducerea incidenei
anaplasmozei (33).
3.2. Hibridarea interspecific n controlul ectoparazitismului. Dintre
metodele de combatere autocidial s-au experimentat pe mai multe specii de
acarieni i insecte parazite sau duntoare. Rezultatele n combaterea cpuelor
ectoparazite sunt favorabile. n experimentri s-au folosit reproducerea ntre
masculi de Boophilus decoloratus i femele de B. microplus, iar ponta acestora a
fost cu ou infecunde. Introducerea masculilor hibrizi n biotopuri populate de B.
decoloratus a provocat n anii urmtori o scdere semnificativ a numrului
populaiei de cpue din aceast specie (32). Hibrizii masculi dintre aceste specii
de Boophilus au fost sterili pn la a treia generaie. Dup doi ani de competiie
ntre masculi s-a observat, n zon, o reducere important a densitii acestei
cpue (25).
Rezultate pozitive s-au obinut i n experimentri cu insecte. Prin cuplarea
masculilor de Hypoderma bovis cu femele de H. lineatum, masculii hibrizi
rezultai au fost infecunzi. Masculi F1 de H. lineatum, obinui n laborator, au
fost dispersai n zone populate cu H. bovis care, n competiie, au fecundat o
parte dintre femele, a cror pont a fost cu ou sterile. Dup 2-3 ani, n biotopurile
respective s-a constatat o reducere important a populaiei i a incidenei
hipodermozei la taurine (2, 3).
Metodele de combatere biologic sunt mult mai diversificate i se refer i
la antagonismul interspecific i intergenic (41), la sterilizarea masculilor,
combaterea feromonal, mijloace genetice .a. (33, 35, 40, 41), depind limitele
unui referat. Aspectele prezentate privind utilizarea i valorificarea paraziilor n
parazitoterapie i combaterea biologic ne conduc la urmtoarele concluzii.

CONCLUZII

n aproape opt decenii de la aplicarea malarioterapiei n premier mondial


(1927), s-au realizat progrese importante care au finalizat cu elaborarea primului
volum de Parazitica (2005), ramur a parazitologiei avnd scop utilizarea
paraziilor n aciuni benefice activitii umane.
Parazitoterapia s-a dezvoltat i diversificat; s-au descoperit parazii i lizate
parazitare cu efecte: oncostatice, antitumorale, antiparazitare, imunosupresoare,
imunomodelatoare, antiinflamatoare .a.
Autodeparazitarea gazdelor, urmat de eliminarea oulor deformate i
nsntoirea pacienilor a fost semnalat n premier n himenolepidoz la noi n
ar.

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n combaterea biologic, folosirea hiperparaziilor a avut efecte pozitive n


controlul neodiplostomozei (metacercariozei), iar hibridarea interspecific a
paraziilor a avut efecte favorabile n controlul hipodermozei i ectoparazitismului
cu cpue.
Cercetrile i experimentrile autohtone au contribuit la: semnalarea n
premier a fenomenelor de autodeparazitare n himenolepidoz la om; aplicarea
hiperparazitismului n controlul diplostomozei petilor; studii de sintez asupra
parazitoterapiei, hiperparazitismului i elaborarea primului volum de Parazitica.

BIBLIOGRAFIE
1. Anene B.M., 1989 - Immunosuppression of humoral immune response in canine
trypanosomiasis. Microbios Letters, 40, 157, 37-46.
2. Boulard C., 1988 - Hypodermose bovine. Point Vtrinaire, 20, 112, 17-24.
3. Boulard C., Argent G., Hellion E., 1988 - Hypodermose bovine, Diagnostic et
traitement. Point Vtrinaire, 20, 112, 105-115.
4. Dorchies Ph., Alzien J.P., 1997 - Loestrose ovine: revue. Rev. Md. Vt. 148, 7,
565-574.
5. Dulceanu N., Terinte C., Mitrea L.I., Polcovnicu C., 2000 - Dicionar enciclopedic
de Parazitologie. Edit. Academiei Romne, Bucureti.
6. Elliott D.E., Urban J.F jr., Arga C.K., 2000 - Does the failure to acquire helminthic
parasites predisposes to Crohns disease? Faber J. 14, 2848-55.
7. Gherman I., 1960 - Dgnrescence des oeufs dHymenolpis nana observe chez
les personnes en voie de gurison spontane. Ann. Parasit. Humaine et
Compare, Paris, V, 35, 5/6, 755.
8. Gherman I., Filoti P., Tutu Lidia, Plecias M., Dresher E., Garofeanu M., Sedroiu
L., 1961 - Dinamica infestrilor i vindecrilor spontane n himenolepidoz.
Igiena, 2, 135-143.
9. Gherman I., Juvara Ana-Maria, 1962 - Semnificaia i importana practic a apariiei
oulor degradate n himenolepidoz cu Hymenolepis nana. Pediatria, 5, 417-
430.
10. Gherman I., 1962 - Dgnrescence spontane des oeufs dHymenolpis nana
forme de manifestation de limmunit de cette parasitose. Ann. Parasit. Humaine
et Compare, Paris, 4, 513-518.
11. Gherman I., 1963 - Despre unele mecanisme patogenetice i unele manifestri
clinice ntlnite n lambliaz. Rev. de Microb. Parazit. i Epid. 4, 353-359.
12. Gherman I., Angelescu C., 1966 - Observations sur la signification pathologique de
la flore bactrienne et de la faune parasitaire de la bile. Revue Internat.
dHepatologie, Paris, 8.
13. Gherman I., 1993 - Compendiu de Parazitologie Clinic, Edit. All, Bucureti.
14. Gherman I., 1994 - Giardiaza (Lambliaza). Observaii de ultim or. Viaa Medical,
51, 23 dec.
15. Gothe R., 1983 - Pheromones in ixodid and argasid ticks. 1. Ixodid ticks. Vet. Res.
Rev. 16-37.
16. Graf J.F., 1975 - Mise en vidence dune pheromone chez Ixodes ricinus.
Acarologie, 17, 436-441.
17. Holmes P.H., Mammo E., Thomson A., Knight P.A., Lucken R., Murray P.K.,
Junnings F.W., Urquart G.M., 1974 - Immunosuppression in bovine
trypanosomiasis. The Vet. Record, 27, 86-87.
18. Iacob N., Lctuu M., Beratlief C., Mihalache G., Ceianu I., 1975 - Combaterea
biologic a duntorilor. Edit. tiinific, Bucureti.
19. Iftimovici R., 1994 - Istoria medicinei. Edit. All, Bucureti.

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20. Khan W.I., Blennerhasset P.A., Varghese A.K., 2002 - Intestinal nematode
infection ameliorates experimental colitis in mice. Infect. Immun. 70, 5931-37.
21. Licperta E., 1958 - Farmacologie veterinar. Edit. Agro-Silvic, Bucureti.
22. Miyahara K., Tose S., Sakurai H., Igarashi I., Saito A., Hirose T., Suzuki N., 1992
- Role of adherent spleen cells in the induction of cytotoxic activity by
Toxoplasma lysate antigen (TLA). J. Vet. Med. Sci., 54, 4, 629-635.
23. Miyahara K., Yoko N., Sakurai H., Igarashi I., Sakata M., 1992 - Antitumor activity
of Toxoplasma lysate antigen against methylcholanthrene induce tumor-Bearing
rats. J. Vet. Med. Sci., 54, 2, 221-228.
24. Ogawa R., Sato M., Hirose T., Suzuki N., 1985 - Preventive effect of Toxoplasma
lysate antigen (TLA) on fatal infection with mouse piroplasma. Jpn. J. Vet. Sci.,
47, 225-236.
25. Osburn R.L., Knipling E.F., 1982 - The potential use of sterile hybrid Boophilus ticks
as a supplemental eradication technique. J. Medical Entomology, 19, 6, 637-644.
26. Oel V., 1991 - Preliminary data concerning the biological control of fish
th
Diplostomosis by hyperparasiting. In: European Assoc. Fish Pathologist. V
Conf., Budapest.
27. Oel V., 1999 - Cercetri asupra diplostomozei la peti n mediul piscicol amenajat i
natural din Delta Dunrii. Tez de doctorat. U.S.A.M.V. Cluj-Napoca.
28. Palmieri J.R., Cali A., Heckmann R., 1976 - Experimental biological control of the
eye fluke, Diplostomum spathaceum, by a protozoan hiperparasite Nosema
strigeoidea. J. Parasitol. 62, 2, 325-326.
29. Sato M., Hori Z., Hirose T., Suzuki N., 1985 - Immune effect of TLA (Toxoplasma
lysate antigen) on cattle against Theileria sergenti infection. Jpn. J. Vet. Sci. 47,
6, 921-926.
30. Silegem M., Darji A., De Baeltselier P., 1991 - In vitro simulation of
immunosuppression caused by Trypanosoma brucei. Immunology, 73, 246-248.
31. Spaldanova R., Oeleny S., Zemberyova A., 1990 - Trichinella spiralis and
th
Trichinella pseudospiralis as inhibitors of tumors growth. VII Congr. Intern.
Parasitol. Bull Soc. Franc. Parasitol. (resumes), 8, 1, 14.
32. Spickett A.M., Malan J.R., 1978 - Genetic incompatibility between Boophilus
decoloratus and B. microplus and hybrid sterility of australian and south african
B. microplus. Anderstepoort. J. Vet. Res., 45, 3, 149-153.
33. Squarzoni C., 2001 - Lutte biologique contre stomoxes des bovins 0020 la reunion:
lexemple de Poseidon Vtrinaire. Thse de doctorat. Ecole Vt. Toulouse.
34. Summers R.W., Elliott D.E., Urban J.F. jr., Thompson R., Weinstock J.V., 2005 -
Trichuris suis therapy in Crohns disease. Gut, 54, 87-90.
35. uteu E., 1993 - Parazitoterapia probleme actuale i perspective. Simp. Actualiti
n patologia animalelor domestice, XX, Cluj-Napoca, 149.
36. uteu E., 1994 - Consideraii asupra hiperparazitismului ca fenomen biologic i
metod de combatere. Rev. Rom. Med. Vet. 4, 2, 121-126.
37. uteu E., 2003 - Combaterea biologic, posibil alternativ a chimiopreveniei unor
helmintoze la animale. Rev. Rom. Med. Vet. 13, 1, 55-60.
38. uteu E., Cozma V., 2004 - Parazitologie clinic veterinar. Vol. I, II, Edit. Risoprint,
Cluj-Napoca.
39. uteu E., 2005 - Parazitica. Edit. Risoprint, Cluj-Napoca.
40. Toma M., Unguruan I., Costea V., Costea Gheorghina, 2003 - Studiu
comparativ privind combaterea duntorului Hyphantria cunea cu insecticide
biologice i insecticide chimice. Rev. Rom. Parasitol. XIII, 1, 103-105.
41. Yusuf J.N., Piekarski G., Pelster B., 1980 - Concurrent infection of Trichinella
spiralis and Toxoplasma gondii in mice. Z. Parasitenkd., 62, 230-240.
42. Weinstock J.V., Summers R., Elliott D.E., 2004 - Helminths and harmony. Gut, 53,
7-9.

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TREATMENT OF ANAEROBIC INFECTIONS


TRATAMENTUL INFECIILOR ANAEROBE

Gh. CRISTEA
C.S.V. uletea
P. CURA, V. NASTASA
U.S.A.M.V. Iai

Although the existence of anaerobic bacteria has known for over a


century, it is only relatively recently that their importance as agents of
disease in both humans and animals has been realized. Our increased
awareness stem largely from improvements in anaerobic bacteriology and
increases in the number of clinical bacteriologic specimens processed for
isolation of anaerobic bacteria. A number of excellent reviews highlighting
the importance of anaerobic infection in persons have appeared over the
past 15 years. Lately, reports of the microbiologic and clinical features of
anaerobic infection in animals have also appeared. Currently, it is accepted
that anaerobic infection occurs frequently in animals, is associated with
distinctive clinical characteristics, and requires specific treatment that in
many instances differs from that required for treatment of aerobic
infections.

Key words: anaerobic infections, antibiotics, therapy, animals

MEDICAL MANAGEMENT OF ANAEROBIC INFECTION

Antimicrobials are generally administered concurrent with surgery for


treatment of anaerobic infection. Unless surgical debridement is very thorough,
infected tissues and bacteria often remain at the original wound site.
Antimicrobials are administered primarily to eradicate the residual bacteria and to
prevent spread into adjacent healthy tissues. Generally, antibiotics are
administered at high doses and for prolonged times because blood supply and
delivery of antibiotics to devitalized tissues is often unpredictable. High
concentrations of bacteria, a characteristic of abscesses, promote synthesis of
enzymes capable of inactivating antibiotics. High serum antibiotic concentrations
create a favorable concentration gradient for penetration of antibiotics into
chronic, often walled-off, sites of infection.
In most instances, selection of antibiotics for treatment of anaerobic
infection is largely empiric. Results of anaerobic culture, because of difficulties
inherent to culture methodology and to isolation and identification of multiple
species of bacteria, are often slow to return. Therefore, initial selection of

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antimicrobials is based on results of cytologic examination of wound specimens


together with clinical suspicion of anaerobic infection. Technical difficulties also
preclude routine determination of antibiotic selection. Therefore, a priori
knowledge of antibiotic spectrum of activity and likely anaerobic bacterial
resistance patterns becomes particularly important for optimal medical
management of anaerobic infection. Awareness of antibiotic tissue penetrability is
also required, especially when treating infections in relatively inaccessible sites
such as the central nervous system and walled-off abscesses. Table 1 lists
antimicrobial useful for treatment of anaerobic infection as well as their predicted
spectrum of activity against important anaerobic pathogens.
Five classes of antibiotics are generally considered to be consistently
effective against anaerobic bacteria, both in vitro and in vivo. Penicillin remains
the antibiotic used most widely for routine treatment of anaerobic infection.
Alternatives to penicillines are usually selected when antibiotic resistance is
encountered or for increased tissue penetration. These antimicrobials include
choramphenicol, clindamycin, metronidazole and cefoxitin.
Penicillins
Penicillins are effective against most anaerobic pathogens, with one notable
exception, and bactericidal, inhibiting cell wall synthesis in both aerobic and
anaerobic bacteria. Excretion of penicillins occurs primarily via a combination
glomerular filtration and renal tubular secretion. Penicillin/induced toxicity,
uncommon in animals, usually resemble acute anaphylaxis. Nausea and vomiting,
which may represent an acute hypersensitivity reaction, are observed relatively
frequently when concentrated solutions of penicillin G are rapidly administered
intravenously. Dilution of the drug concentration and slowing the infusion rate
usually resolve these signs.
Penicillin G is the prototype penicillin for treatment of anaerobic infection.
For treatment of severe infections, penicillin G should be administered
intravenously in order to achieve effective serum concentrations. For clostridia
infections and infections caused by anaerobic streptococci, penicillin G remains
the drug of choice. Penicillin G also has good activity against most species of
Actinomyces and Fusobacterium and some species of Bacteroides. However,
several species of Bacteroides, especially B. fragilis, are uniformly resistant to
penicillins. In the case of B. fragilis, production of -lactamases is
chromosomally mediated. Penicillin resistance is less predictable in other strains
of Bacteroides but has been observed intermittently in nearly all, including two
important veterinary pathogens, B. melaninogenicus and B. assacharolyticus.
Bacteroides resistance to penicillins appears to be increasing, both in veterinary
and human isolates. B. fragilis is isolated relatively infrequently from animals,
suggesting that penicillin resistance may also be relatively common amongst non-
fragilis isolates. Because antibiotic resistance in Bacteroides is also conferred by
plasmid transfer, antibiotic selection pressure is probably responsible for much of

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the increase in penicillin resistance. Resistance to penicillins also appears to be


increasing among some strains of Fusobacterium and Clostridium isolated from
humans. Several strains of Fusobacterium resistant to penicillin have also been
isolate recently from cattle.
The anaerobic spectrum of activity of aminopenicillins (ampicillin,
amoxicillin) closely parallels that of penicillin G. Because amoxicillin achieves
consistently higher serum concentrations, it generally is preferred over ampicillin
for both parenteral and oral administration. Amoxicillin given orally is more
completely absorbed than ampicillin, resulting in less disruption of normal
gastrointestinal bacterial flora. Antistaphylococcal penicillins, such as oxacillin
and cloxacillin, are considered less effective than penicillin G against most
anaerobic bacteria. Expanded spectrum penicillins, including carbenicillin and
ticarcillin, may in some instances be more effective against B. fragilis but in
general offer little advantage over other penicillins for treatment of anaerobic
infection. Newer broad-spectrum penicillins (for example, azlocillin, piperacillin,
mezlocillin), although very effective against Pseudomonas and other gram-
negative bacilli, are roughly equivalent to ticarcillin and carbenicillin in activity
against anaerobic bacteria, including B. fragilis.
One attempt to expand the antibacterial spectrum of penicillins has been to
combine them with -lactamase inhibitors, since -lactamases mediate most
bacterial resistance to penicillins. Clavulanic acid, which by itself has little
antibacterial activity, has been used most often for this purpose. When amoxicillin
is combined with potassium clavulanate, its antibacterial spectrum is expanded to
include penicillinase-producing staphylococci and many strains of bacteria in the
Enterobacteriaceae family. Activity against obligate anaerobes, including most
strains of B. fragilis, is improved significantly.
Clindamycin
A lincosamide antibiotic1, clindamycin is primarily bacteriostatic, exerting
its antibacterial effect by binding to the bacterial 50S ribosomal subunit and
inhibiting synthesis of peptide bonds. Clindamycin is well/absorbed orally,
reaching peak serum concentrations 2 to 4 hours after administration. Tissue
concentrations of clindamycin generally exceed serum concentrations. Effective
therapeutic concentrations are achieved in most body tissues, including bone and
joints, with the exception of the central nervous system, where concentrations are
low. At least 60 per cent of clindamycin undergoes hepatic metabolism and
biotransformation, producing some metabolites that are more active than the
parent compound. Some active clindamycin is excreted unchanged in urine,
although biliary excretion is the primary route of elimination. For these reason,
clindamycin should be used cautiously in patient with hepatic dysfunction.
Pseudomembranous colitis (a potentially fatal disease of the large bowel caused
by antibiotic-induced colonic overgrowth of cytotoxic strains of Clostridium
difficile).

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An important characteristic of clindamycin is its ability to penetrate to


effective concentrations in walled-of abscesses, surpassing concentrations
achieved by florphenicol and penicillins. In addition, clindamycin is selectively
concentrated within leukocytes by an energy-dependent process, reaching
concentrations 5 to 50 fold greater than serum concentrations. This has important
implications for treatment of chronic, persistent infections characterized by
intracellular localization of bacteria. Bacteria sequestered within leukocytes are
protected, since few antibiotics, with the exception of clindamycin, rifampin, and
to a lesser degree, florphenicol are capable of penetrating into white blood cells.
Berg and coworkers determined experimentally that clindamycin was more
effective than its parent compound, lincomycin, for treatment of anaerobic
infections n cows.
Metronidazole
Metronidazole2 has excellent activity against many anaerobic bacteria.
Reduction of metronidazole`s nitro group by susceptible bacteria possessing
nitroreductases results in formation of short-lived, highly reactive intermediates
that disrupt bacterial DNA. This reaction precedes best under low redox
conditions (typical of anaerobic infection) in susceptible bacteria possessing low
redox electron transport proteins. Under these conditions, electrons are readily
available for nitro reduction; under aerobic conditions, oxygen competes for
electrons, inhibiting the reduction reaction. Onset of antibacterial activity is
abrupt, as opposed to the more gradual onset of action of penicillin and
clindamycin.
Metronidazole is very well absorbed orally, with a serum half-life of 3 to
13 hours; bioavailability after oral administration cows ranges from 50 to 100 per
cent. Tissue distribution is quite wide; effective concentrations are attained in
bone, bile, cerebrospinal fluid, and brain parenchyma. Metabolism occurs
primarily in the liver and major routes of excretion are via urine and feces.
Metronidazole should be used cautiously in animals with hepatic disease.
Adverse reactions (largely gastrointestinal disturbances) from
metronidazole are uncommon in humans. Occasional neurologic abnormalities,
including peripheral neuropathy, seizures, and ataxia, have been observed in
human patients receiving high doses of metronidazole or prolonged treatment.
Previously, few adverse effects were associated with administration of
metronidazole to cows. Recently, however, serious neurotoxicity has been
observed both clinically and experimentally in a number of cows treated with high
doses of metronidazole. In a few cases, doses administered were close to a
currently recommended dose, 66 mg per kg per day. Onset of neurologic signs
was usually acute, often preceded for 12 to 24 hours by anorexia. Characteristic
neurologic abnormalities have included severe ataxia, vertical or rotatory
nystagmus, and occasionally seizures. After drug discontinuation, most cows have
slowly recovered over a period of 1 to 2 weeks.

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Metronidazole is unique in that it is the only antimicrobial currently


available with consistent bactericidal activity against B. fragilis and most other
strains of Bacteroides. In addition, metronidazole is active against other gram-
negative and gram-positive spore-forming anaerobes. Activity is poor against
Actinomyces, Propionobacterium, and microaerophilic streptococci (technically,
not anaerobes). Metronidazole is generally considered to be completely inactive
against aerobic bacteria, though there is evidence to suggest that metronidazole,
when used to treat experimentally created mixed infections, has activity against
coliform bacteria; anaerobic bacteria may convert metronidazole to intermediates
active against coliform bacteria. Effective tissue penetration and consistent
bactericidal activity make metronidazole the drug of choice for severe anaerobic
infections, particularly those involving Bacteroides. Specific indications for use
of metronidazole include anaerobic osteomyelitis, anaerobic endocarditis,
anaerobic brain abscesses, and intra-abdominal abscesses that cannot be
completely eradicated surgically.
Cephalosporins
Cephalosporins3 are a broad class of bactericidal antibiotics possessing a -
lactam ring and exhibiting antibacterial activity similar to that of penicillins. They
are divided into three categories (generations) based on time of development,
structural characteristics, and spectrum of activity. In general, first-generation
cephalosporins are most active against gram-positive bacteria, whereas third-
generation cephalosporins have increased activity against gram-negative
pathogens and reduced effectiveness against gram-positive bacteria. Most
cephalosporins, with the exception of cefaperazone (biliary excretion), are
eliminated primarily via the kidneys. Adverse reactions are relatively uncommon
and tend to vary widely amongst various cephalosporins.
Cephalosporins are relatively active against most species of anaerobic
bacteria except B. fragilis. This is especially true of first-generation
cephalosporins (cephalothin, cephapirin, cefazolin, cefadroxil). Isolates of
Bacteroides from domestic animals have been shown to have high rates of
resistance to first-generation cephalosporins. Bacteroides resistance to
cephalosporins is mediated by a constitutive -lactamase. On the other hand,
cefoxitin, a cephamycin antibiotic classified as a second-generation
cephalosporin, is -lactamase resistant and is effective against most strains of B.
fragilis. Cefoxitin is currently considered the most effective cephalosporin for
treatment of anaerobic infections, especially those caused by Bacteroides. Broad-
spectrum activity, including activity against multiresistant strains of bacteria in
the Enterobacteriaceae family, is another advantage of cefoxitin. For these reason,
cefoxitin is considered useful for monotherapy of mixed infections containing
obligate anaerobes and coliform bacteria. In the authors experience, cefoxitin has
been effective in treatment of mixed infections of bone and lungs in cows.

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Imipenam
Imipenam5, the first of a new class of -lactam antibiotics known as
carbapenams, has significant activity against most medically important species of
bacteria, including multiresistant strains of Pseudomonas. Imipenam undergoes
extensive renal tubular metabolism and is therefore combined with cilastatin, an
inhibitor of renal tubular depeptidase. The combination results in much higher
urine concentrations of imipenam and also reduces nephrotxicity associated with
administration of imipenam alone.
Activity of imipenam against anaerobic bacteria exceeds that of all other -
lactam antibiotics and is considered equivalent to the activity of clindamycin and
metronidazole. Obviously, a potent broad-spectrum, relatively nontoxic antibiotic
such as imipenam offers important advantages for treatment of serious mixed
infections, particularly when culture and susceptibility information is not known.
Although it has good efficacy in human beings, there has been little clinical
experience with this antibiotic in veterinary medicine.
Antibiotics with Limited Effectiveness Against Anaerobes
Anaerobic bacteria are uniformly resistant to aminoglycoside antibiotics4,
due to the fact that transfer of aminoglycosides across the bacterial cell membrane
requires enzymes that are lacking in anaerobic bacteria.
Polymyxin antibiotics are also ineffective, for reasons similar to those for
aminoglycosides.
Widespread resistance to tetracycline has greatly limited its usefulness in
treatment of anaerobic infection. Hirsch and colleagues found tetracycline
resistance in a number of Bacteroides isolates from veterinary patients.
Sulfonamides also have limited efficacy for treatment of anaerobic
infection because in environment typical of anaerobic infection, by product of
cellular breakdown, including para-aminobenzoic acid, are present in abundance.
Many of these substances competitively inhibit the activity of sulfonamides.
Trimethoprim was also shown to be clinically ineffective against anaerobic
bacteria. Sulfonamide with trimethoprim performed poorly in an anaerobic
infection treatment model and failed to reduce abscesses formation or produce
cures. Despite evidence of in vitro activity against obligate anaerobes isolated
from animals, trimethoprim-sulfamethoxazole was found clinically to be an
ineffective treatment for anaerobic infections in calves and foals and was
associated with a significant number of treatment failures.

SUMMARY

Successful management of anaerobic infection first requires an accurate


diagnosis. Cytologic examination of wound exudates and inspection for
characteristic clinical clues greatly facilitates an accurate initial diagnosis of
anaerobic infection. Knowledge of antimicrobial activity against specific

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anaerobic pathogens is essential, since antibiotic susceptibility information is not


routinely available. Whenever possible, antimicrobial and surgical therapy should
be combined in managing anaerobic infections.
Florphenicol, clindamycin, and metronidazole provide the most
consistently reliable activity against pathogenic anaerobes, including Bacteroides.
Penicillins are also generally effective, except for treatment of infections caused
by penicillinase-producing strains of Bacteroides. Cephalosporins are not
considered drugs of choice for anaerobic infections, although cefoxitin may in
some instances be useful as monotherapy of mixed infections containing obligate
anaerobes and coliform bacteria. Aminoglycosides and sulfonamides are
ineffective and should be avoided for treatment of anaerobic infection.

REFERENCES

1. Berg, J.N. et al.,1994 Clical models for anaerobic bacterial infections of cows and
their use in testing the efficacy of clindamycin and lincomycin. Am. J. Vet. Res.,
40:1299-1306.
2. Nastasa, V, 2005 - Farmacologie veterinar I, 145-170
3. Papich, M.G., 2004 Clinical pharmacology of cephalosporin antibiotics. J. Am. Vet.
Med. Assoc., 184:344-347
4. Pechere, J.C., Delisk, R., 1993 Early selection of antibiotic in anaerobic infections.
Surgery, 93:134-144.
5. Tally, F.P., Jacobus, N.V., 2003 Susceptibility of anaerobic bacteria to imipenem.
J. Antimicrob Chemother 12 (Suppl D): 47:51.

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CLASSIC USE OF LOCAL ANAESTHETICS IN A


BALANCED ANAESTHESIA PROTOCOL
UTILIZAREA ANESTEZICELOR LOCALE CLASICE N PROTOCOLUL
DE ANESTEZIE ECHILIBRAT

N. MUNTEANU1, P. CURA2,
Gh. CRISTEA1, V. NASTASA2
1
D.V.S.A. Vaslui
2
U.S.A.M.V. Iai

A combination of local or locoregional anaesthesia with general


anaesthesia has several very important advantages: 1) A satisfactory
surgical plane of anaesthesia can be obtained with much lesser doses of
general anaesthetics (volatile or intravenous agents). This means less
cardiovascular and respiratory depression, a more stable anesthesia,less
metabolisation efforts, quicker recoveries. 2) The phenomenon know as
"winding-up" of the central nervous system (CNS) will be at least less
pronounced. Winding up results in a prolonged sensitization of the CNS and
hyperalgesia. With the combination of the local techniques and a general
(light) anaesthesia the postoperative analgesia will be better/longer or at
least easier to control with additional medication. Complications are few if
the recommended maximum dosis of the local anaesthetics are respected.

Key words: local anaesthetics, analgesia, general anaesthesia.

The advantages of this philosophy cannot be overestimated and are


particularly attractive and easy to apply in practice. The classic agents are the
more short acting (1-2 hrs) lidocaine (1% to 2%)1 with a fast (5 min) onset and the
longer (4-6 hrs) acting bupivacaine2 (0.25%-0.7%) with a slow (20 min) onset.
Both agents can be mixed together.
The possibilities include: a) topical application; b) tissue infiltration; c) ring
block; d) nerve blockade; e) intravenous regional anaesthesia; f) spinal
anaesthesia (epidural, subdural).
Off all these techniques topical application, tissue infiltration, ring blocks
are very easy to do but their potential benefit is often unrecognised for reasons
difficult to explain. Intravenous regional anaesthesia is also an easy procedure.
Epidural anaesthesia is in the reach of each practitioner.

1. Topical application
Even if a general anaesthesia procedure is done, noxious output from the
operated area can be reduced by application of local anaesthetic ointments, liquid
solutions or sprays (lidocaine)3. Remember that the intact skin is not penetrated
except following application of a special formula of prilocaine (EMLA).
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Application on nasal and oral mucous membranes, esophagus, trachea and


urogenital mucosae is possible. Straightforward simple applications during
surgical procedures are e.g., splash of lidocaine on the spermatic cord during male
castration and on the ovarian pedicle during ovariectomy. Local anaesthetics do
not retard wound healing and an increase in the tendency to diffuse bleeding has
not been documented as a problem.

2. Infiltrations/ring block
These remain a very simple but often neglected possibility. Aspiration
before injection will limit the risk for intravascular injection. Examples of a
possible use are e.g., removal of small skin tumors, suturing skin wounds, surgery
on the toes (distal phalanx amputation, interdigital wounds or cysts, wounds etc),
incisional infiltration of subcutis and fascia before a caesarean or a laparotomy in
critical cases.

3. Intravenous regional anesthesia (Bier's block)


Although a simple and effective technique for distal extremity surgery not
exceeding 1.5 hrs this anaesthetic technique is not always favoured by some
surgeons because of the lack of bleeding in the operated area. It can be very useful
for relatively minor surgery on the extremities of critical or debilitated patients.
Using a tourniquet and/or cuff combined with intravenous lidocaine an effective
analgesia after 5-10 minutes in all tissues distal to the tourniquet/cuff can be
obtained, sensibility returns within minutes after release of the tourniquet or cuff.

4. Plexus Brachialis block


This is an excellent technique to be used, e.g., for radius/ulna surgery. It is
a block of the n. axillaris, n. medianus, n. ulnaris, n. musculocutaneus, n. radialis
following successful infiltration of the area of the plexus brachialis situated at the
level of the shoulder joint at the medial side of the scapula. The exact quantities of
local anaesthetic to use are not described and depend on the size of the patient and
the toxic dosis (e.g., 2-20 ml lidocaine 1 or 2% mixture). Electrolocation of the
plexus with a nerve stimulator is a useful way to obtain maximum block with a
small amount of local anaesthetic. If successful there will be analgesia and
motoric paralysis (not appreciated when this block is performed under
anaesthesia). Analgesia extends from the elbow to distal. Successful block will be
apparent when conducting general anaesthesia: low concentration of inhalation
agent necessary, stable vital parameters. Complications are rare but
pneumothorax, nerve trauma, bleeding and intravascular injection are possible.

5. Intercostal blocks
These are used before the start of a thoracotomy (most logic approach for
"preventive") or when starting to close the thorax; the technique can also be used

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for analgesic support when managing rib fractures. The technique consists of:
injection in the intercostal muscles at the level of the caudal aspect of the rib close
to its origin 0.2 to 1 ml 2% lidocaine (or bupivacaine, or a mix of bupivacaine and
lidocaine) and this in two intercostal spaces before and two spaces after the
thoracotomy incision; this can be done from outside or from inside the thorax.

6. Spinal anesthesia
Mostly used is epidural anesthesia. Accidental puncture of the duramater
and collection of cerebrospinal fluid however is possible when attempting an
epidural injection. This will happen more often in the cat were the dural sac is
longer than in the dog. The combination of an epidural anaesthesia and general
anaesthesia is a very good technique for surgery caudal to the ribs (orthopedics,
soft tissues, and caesareans). It allows a light level of general anaesthesia, a stable
plane of anaesthesia, and post operative pain relief. In general local anaesthetics
are used but they can be combined with opioids.
The local anaesthetic can be combined with an opioid for epidural
injection. They will have an interesting and practical synergistic analgesic effect.
This way a good and prolonged postoperative analgesic effect can be obtained.
Addition of 0.1 mg/kg methadone or morphine can give up to 10 hours of
postoperative analgesia (motoric effects having disappeared).
Local anaesthetics offer the opportunity to obtain prolonged postoperative
pain relief by using a long acting preparation like bupivacaine, by repeating the
described applications (e.g., re-doing an epidural), by combining with other drugs
(e.g., epidural with local anaesthetic and an opioid), or by continuous infusion in
the epidural space via an epidural catheter. Incisional and intraabdominal
application of lidocaine preceding closure of laparotomy wounds has been
described in human and in dogs as a mean of postoperative analgesia.

INTRAVENOUS LIDOCAINE IN BALANCED ANAESTHESIA


PROTOCOL AND POSTOPERATIVE PAIN MANAGEMENT

Intravenous lidocaine has been shown to reduce the minimum alveolar


concentration (MAC) of isoflurane (and halothane) in several species including
dogs. For this reason it can be used in a balanced anaesthesia protocol for critical
patients. Furthermore it has distinct analgesic properties and can be used as and
adjunct in iv infusion to manage difficult cases with postoperative pain; it is
claimed particularly useful in protocols for visceral pain and for more chronic
pain. Dose iv: 1.5-3 mg/kg followed by a loading dose of 5-200 mcg/kg/min.

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SUPPLEMENTARY USEFUL ASPECTS


OF LIDOCAINE THERAPY

Lidocaine as an antiarrhythmic1 agent is classically used to treat ventricular


extrasystoles. Dogs presented with gastric volvulus often show these rhythm
disturbances. Lidocaine therapy can both treat/prevent ventricular extrasystoles
and offer at the same additional analgesia during and after surgery. There is also a
large body of evidence from animal experiments that supports the potential of
lidocaine as an anti-oxidant and inflammatory modulator useful in preventing
reperfusion injury. Therefore it seems attractive to consider it for the treatment/
prevention of systemic inflammatory response (SIRS) and multiple organ
dysfunction syndrome (MODS). Although conclusive clinical evidence is lacking
lidocaine infusions are also used for their prokinetic properties to prevent/treat
postoperative ileus (human, horses)4. Propulsive activity at the level of the
proximal duodenum is enhanced.

REFERENCES

1. B.H. Cassuto, R.W. Gfeller, 2003 - Use of intravenous lidocaine to prevent


reperfusion injury and subsequent multiple organ dysfunction syndrome, J vet
emergency and critical care 13 (3), 2003, 137-148
2. R.E. Carpenter, D.V. Wilson, A.T. Evans, 2004 - Evaluation of intraperitoneal and
incisional lidocaine or bupivacaine for analgesia following ovariohysterectomy.
Veterinary Anaesthesia and Analgesia 31 (1), 2004, 46-52.
3. L.J. Smith, E. Bentley, A. Shih, P.E. Miller, 2004 - Systemic lidocaine infusion as
an analgesic for intraocular surgery in dogs: a pilot study Veterinary Anaesthesia
and Analgesia 31 (1), 2004,53-61
4. P. Brianceau, H. Chevalier, A. Karas, M.H. Court, L. Bassage, C. Kirker-Head, P.
Provost, M.R. Paradis, 2002 - Intravenous lidocaine and small-intestinal size,
abdominal fluid and outcome after colic surgery in horses J Vet Intern Med 16,
736-741

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PHARMACOCLINICALS CONSIDERATIONS
ON THE EVOLUTION OF CARDIOVASCULARES
VARIABLES AND ACID-BASE BALANCE
UNDER THE COMBINATION ACEPROMAZINE-
BUPRENORPHINE IN DOG
CONSIDERAII FARMACOCLINICE ASUPRA EVOLUIEI VARIABILELOR
CARDIOVASCULARE I A ECHILIBRULUI ACIDO-BAZIC SUB COMBINAIA
ACEPROMAZIN BUPRENORFIN LA CINE

V. NASTASA1, P. CURA1, N. MUNTEANU2,


FL. CURA1, Mariana GRECU1
1
U.S.A.M.V. Iai
2
D.V.S.A. Vaslui

The objective of this paper was to appreciate the intensity of the


modification of principals variables heart-respirators under the
anaesthesia with a combination between acepromazine - buprenorphine at
dog.

Key words: anesthesia, acepromazine, buprenorphine, dogs.

MATERIAL AND METHOD

The researchers were realised on 16 dogs healthiness from an ordinary


breed. Before starting the experiment on the body of the dog was introduced a
catheter intravenous in the femoral artery and cephalic vein previous brachial for
prelevation the proved of blood for analysis the pressures of blood gases (PaCO2,
PaO2, PvCO2, PvO2), pHa, pHv, HCO-3a, HCO-3v using Compact 2 AVL analyser.
To measuring the arterial blood pressure was used an electronic apparatus
noninvasive (made in Tokyo, Japan). The cardiac frequency was appreciated
using a precordial stethoscope. The respiratory frequency was determinate by
measurement of "trip of thorax".
Acepromazine (Vetranquil, solution 1%)2 was administered after the
recording the basic values in dosage 0.4 mg/kc intravenous during over than 30
seconds. The measurement of cardiorespiratory variables was repeated at 15
minutes after the administration of acepromazine. Immediately after the realising
the second set of measurements was administered intravenous the first dose of
buprenorphine1, respectively 0.02 mg/kc during the same interval by 30 seconds.
The measurements of cardiorespiratory variables were realised afterwards 15
minutes. The second dose of buprenorphine respectively 0.18 mg/kc was
administered after measurements the total doses cumulate were 0.2 mg/kc. The
measurements were realised after 15 minutes that the injection was done.

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The scale used to appreciate the level of sedation was imagined after the
degree of conscious and maintenance of laterally recumbency. So, was noted
with 0-dog conscious, 1-dog semiconscious and laterally recumbency inconstant
and 2-dog unconscious, laterally recumbency permanent.
The statistic evaluation was realised using the ANOVA test (variance
analysis) for repeated measurements. So, for a value of P<0.05, the results were
considerate significant. The data are presented like X SX.

RESULTS AND DISCUSSIONS

The level of sedation was significant more high after the administration the
combination acepromazine-buprenorphine comparatively with the bases values.
This is present in a rapport by 23% after the administration of acepromazine (0.5
mg/kc the level 1st), next in percent by 89% after the administration intravenous
of the fist dose of buprenorphine (0.01 mg/kc, the second level) and into a percent
by 100% after the administration of the next dose of buprenorphine (0.2 mg/kc,
the second level). Instead, was recorded an important reducing of respiratory
frequency into a ratio by 59% after the administration intravenous of
acepromazine (Table 1).
Table 1
The evolution of modifications of cardiorespiratory variables and blood gases
in 16 dogs after administration the combination acepromazine-buprenorphine

Values for ACE Buprenorphine Buprenorphine ANOVA


Variables
references (0.4 mg/kc) (0.02 mg/kc) (0.2 mg/kc) P value
PAS (mm/Hg) 154 15 118 12 117 8 113 6 < 0.001
PAD (mm/Hg) 92 14 75 11 * 72 10 65 10 < 0.001
PAM (mm/Hg) 120 13 97 12 * 90 9 88 9 < 0.001
FC (beats/min.) 98 14 96 21 80 25 69 18 < 0.01
FR
48 35 25 11 * 23 22 17 13 < 0.01
(respirations/min.)
pHa (U) 7.42 0.02 7.42 0.02 7.41 0.04 7.40 0.02 < 0.001
PaCO2 (mm/Hg) 31.8 3.0 28.8 4.7 33.0 3.2 * 33.9 3.7 < 0.01
PaO2 (mm/Hg) 87.8 5.9 90.0 90.2 87.9 5.4 90.9 9.8 NS
H2CO-3a (mEq/l) 21.1 2.0 19.0 2.6 19.6 1.9 21.0 1.7 NS
pHv (U) 7.40 0.01 7.40 0.01 7.38 0.02 * 7.38 0.02 * < 0.001
PvCO2 (mm/Hg) 34.9 3.6 34.6 2.8 38.3 2.5 * 39.6 2.9 * < 0.001
PvO2 (mm/Hg) 41.0 3.8 41.0 5.0 40.9 4.0 40.7 2.3 NS
H2CO-3v (mEq/l) 21.8 1.5 21.0 1.6 21.5 1.5 22.4 1.5 * < 0.05

*-the significant difference (P<0.05) versus the value immediately previously. The data are
presented like X Sx; PAS-the systolic heart pressure; PAD-the diastolic heart pressure;
PAM-the media heart pressure; FC-the heart frequency; FR-the respiratory frequency;
pHa-the arterial pH; PaCO2-the tension of CO2 arterial; H2CO-3a-the concentration in
bicarbonate arterial; pHv-the venous pH; PvCO2-the tension of CO2 venous; PvO2-the
-
tension of oxygen; H2CO 3v-the concentration in venous bicarbonate; NS-insignificant.

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The majority of variables modifications of haemodinamic function are


recorded after the administration of acepromazine. Although, the media heart rate
are reduced into a ratio by 11 % after the administration of acepromazine. This
value doesnt significant different versus the basis values. Also, the arterial
pressure was reduced after the administration of the acepromazine (ACE).
Buprenorphine doesnt induced modifications significant statistic in
variables of haemodinamic function excepting the heart rate that recorded a
reducing with 7% after the administration the high level of buprenorphine (Table
1).

CONCLUSIONS

The results of these experiment proved that the association between


acepromazine-Buprenorphine in a dose clinical underlined it is well tolerate from
point of view of haemodinamic function. The reducing of the data recorded in the
heart pressure had media intensity and could be attributed to draining of
simpatico-activity. This activity could be growled during the period of
measurements of the basic values and after the administration of acepromazine.
Also, the aditioning of Buprenorphine into a dosage until 10 times over than
clinical usually dosage are determinate the obtaining of an important sedative
effect without an essential damage of heart-respirators variables at heartless dogs
unpreanesthesiated. A range of dosage varied between 0.2-0.5 mg/kc
acepromazine associated with 0.02-0.2 mg/kc buprenorphine administered
intravenous could be considered satisfactory after the results obtained in our
experiments.

BIBLIOGRAPHY

1. Short, C.E., 1987 Pain, analgesics, and related medications. In: Short CE, ed.
Principles and practice of veterinary anaesthesia. Boltimore: Williams & Wilkins
Co, 28 46.
2. Both, N.H., 1982 Neuroleptanalgesics, narcotic analgesics and analgesic
antagonists. In: Booth N.E., McDonald LE, eds. Veterinary pharmacology and
therapeutics. 5th ed. Ames, Iowa: Iowa State University Press: 267 - 296.

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EVALUATION OF ETOMIDATE AND ALFENTANYL


IN DOG ANAESTHESIA
EVALUAREA ANESTEZIEI CU ETOMIDAT I ALFENTANIL LA CINE

V. NSTAS, Maria-Magdalena NECHIFOR,


Mariana GRECU
U.S.A.M.V. Iasi

The usefulness of etomidate2 for short periods of anaesthesia (e.g.


perform biopsy, skin suture, radiography) is related both to its short
duration of action and the absence of significant side - effects.
The aim of this study has been to monitor the course of anaesthesia
using a combination of etomidate and alfentanyl1.

Key words: anaesthesia, etomidat, alfentanyl, dog.

MATERIALS AND METHODS

30 apparently healthy dogs were studied. They were allocated to three


groups. Dosage regimes for etomidate and alfentanil were respectively: group 1
(n = 22): 1 mg/kg etomidat and 4 g/kg alfentanyl; group 2 (n = 20): 1 mg/kg
etomidat and 8 g/kg alfentanyl; group 3 (n = 18): 1,5 mg/kg etomidat and 4 g/kg
alfentanyl.
Doses were administered by intravenous route. For each group, the heart
rate and respiratory frequency, the level and duration of muscle relaxation and
analgesia were recorded. Assessment of muscle relaxation was made
qualitatively. It was graded as deep when the animal was immobile and moderate
when some spontaneus movements (tail, legs) were observed.
Analgesia was considered to be marked when no response was evoked by
a painful stimulus (prick with a needle) and when the interdigital reflex was absent
Analgesia was considered to be moderate when the interdigital reflex was again
recorded. In addition, rotation of the eyeball and the times between administration
and the first righting of the head and between administration and the first attempt
to walk were measured.

RESULTS AND DISCUTION

Results for relaxation, analgesia and recovery parameters are summarized


in Table 1. When the 30 anaesthetized dogs were considered, 4 cases of excitation
were observed, which necessitated and additional ketamine administration (3,5
mg/kg). Most of the excitated dogs were very nervous prior the induction of
anaesthesia as a results of prolonged travel. In all three groups, bradycardia
occurred after the first minute postinjection: in the first group, return to control

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values occured before the end of anaesthesia (13th minute). When marked
bradycardia occurred, several dogs displayed arrhytmias. Other side-effects were:
vomiting (n = 1) and recovery excitation (n = 3).
This study confirm previously reported data. The normally recommended
dosage regim (group 1) is generally sufficient for most purposes. In 9,5% of the
cases, doses of etomidate had to be increased by 50 %. This combination of drugs
produces only slight cardiovascular effects and has the shortest duration of action.
In contrast, when the dose of fentanyl is doubled, analgesia is prolonged by 4 min
but the depressant effects on the cardiovascular system become a limiting factor.
The dose levels used in the third group led also to undesirable side-effects (the
depressant effect on the cardiovascular system is the most important and, in
addition, awakening is delayed).
Tabel 1
Respose (duration in minutes or minutes after administration) of dogs to the
administration of a combination of etomidate and alfentanyl

1 2 3
Groups
(n = 22) (n = 20) (n = 18)
Etomidate (mg/kg) 1,0 1,0 1,5
Alfentanyl (g/kg) 4,0 8,0 4,0
Muscle relaxation
Deep 84 73 92
Moderate 11 3 12 2 16 1*
Analgesia
Marked and moderate 98 14 1* 15 3*
Eyball rotation 4 5 67 7 5*
Awakening 11 4 104
First righting of the head 12 9 14 7 19 1*
First attempt to stand 17 2 19 6 25 3*
Walking 21 9 22 8 30 6*
Values are expressed as mean SD
* Significant (p < 0,05) with group 1 as control

CONCLUSSION

The main advantages of etomidate alfentanil combination are the


following:
- a short duration of action;
- sufficient analgesia for minor surgery;
- few side-effects on the cardiovascular system which makes this
combination useful in cases of shock.
The main disadvantages are:
- excitation in about 10% of dogs which necessitates ketamine
administration;

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- the occurrence of panting, which makes radiographic examination


difficult especially in the large dogs;
- the volume of the administered dose is important (6,5 ml/kg) and
necessitates and intravenous catheter;
- the combination s expensive.

REFERENCES

1. Arndt J.O., Bednarski B., Parasher C., 1986 Anesthesiology 64, 345.
2. Janssen P.A.J. et al., 1975 Archives of International Pharmacodynamics and
therapeutics 214, 96.

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OBSERVAII PRIVIND EFECTUL UNOR MEDICAMENTE


ANTIPARAZITARE ASUPRA PARAMETRILOR
HEMATOLOGICI LA OVINE CU FASCIOLOZ
ON THE EFFECT OF SOME ANTIPARASITIC DRUGS ON THE HAEMATOLOGICAL
PARAMETERS IN OVINES WITH FASCIOLIASIS

E. PURICE
C.S.V. Vidra, jud.Vrancea
P. CURA, Geta PAVEL
U.S.A.M.V. Iai

In order to apply an antiparasitic therapy with different drugs, from


a herd of ovines infested with Fasciola hepatica, a number of seven flocks
(n=35) were made up. To establish the degree of the anaemia reduction and
of the influence on leukocytemia and trombocytemia, before administration
and 30 days after therapy, haematological measurements were performed.
Results of the research showed up the fact that the benzimidazolic
substances (Fasciocid Triclabendazol 10%; Ovizol 300 Albendazol
300mg/bolus) had the highest efficiency, implying a complete reduction of
anaemia (leukopenia and trombocytopenia have been observed as well).
Also, it has been observed that the same substance (Albendazol)
administrated as bolus (Ovizol 300) is more efficient than when its
administrated as a drinkable solution (Helmizol 2,5). The compound
Fascioxamid has diminished neither the anaemia degree nor the leukocyte
count. Levamisol has not reduced the anaemia degree, but due to its
immunomodulator effect, determines the restoration of the physiological
leukocytemia.

Key words: fascioliasis, sheeps, benzimidazolic therapy, anaemia, leucopenia,


trombocytopenia

In zonele cu ploi abundente, datorit imposibilitii de eradicare a limneelor


(gazdele de intermediare ale fasciolelor), infestrile i reinfestrile conduc la
instalarea fasciolozei cronice, manifestat prin anemie, cahexie, colangit
hiperplazic i ciroz hipertrofic cu acumulare de lichid ascitic, pierderile
economice fiind importante (3, 9). In aceste condiii, este necesar utilizarea unor
medicamente care s distrug att fasciolele adulte, ct i formele tinere aflate n
migraie prin parenchimul hepatic ctre canaliculele biliare (11).
Eficiena medicamentelor antiparazitare fasciolocide este variabil,
dependent de substana activ, concentraia acesteia, timpul de aciune a
medicamentului i mecanismul de aciune asupra paraziilor. Substanele
antiparazitare fasciolocide produse n ultimii ani sunt mai puin toxice i au
eficacitate crescut asupra fasciolelor n toate stadiile de dezvoltare (9).

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MATERIAL I METOD

Intr-un efectiv de ovine dintr-o regiune cu terenuri fasciologene (Cdimani,


judeul Vrancea) s-a diagnosticat clinic i s-a confirmat morfopatologic fascioloza.
In luna octombrie, 2005, oile au fost supuse terapiei cu diverse substane
antiparazitare fasciolocide, urmrindu-se gradul de reducere a strii de anemie i
influena asupra leucocitemiei i trombocitemiei. Substanele antiparazitare
utilizate i modul lor de administrare sunt prezentate n tabelul 1 (2, 5, 9, 11).
Animalele tratate au fost grupate n 7 loturi de cte 5 animale (35 oi): 6 loturi au
primit medicaie antiparazitar diferit, iar un lot a constituit martorul, nefiind
supus terapiei antiparazitare.
Inainte de admnistrarea medicamentelor i la interval de 30 zile dup
terapie, de la animalele investigate s-au recoltat probe de snge pentru evaluarea
eritronului (numrul de eritrocite, concentraia de hemoglobin, hematocritul), a
leucocitemiei i trombocitemiei (4, 6, 7, 8, 10).
Tabelul 1
Substanele antiparazitare administrate la ovinele cu fascioloz investigate

Stadiile
Nr. Preparatul Substana Forma i modul
Doza folosit parazitare
crt. administrat activ de administrare
distruse
1. Fasciocid Triclabendazol Soluie buvabil, 5 ml/ 50kg g.v. ou,
10% per os fasciole
tinere,
adulte
2. Helmizol 2,5 Albendazol Suspensie, 3 ml/10 kg g.v. trematode
2,5% per os adulte
3. Ovizol 300 Albendazol Boluri, 2 boluri/animal trematode
0,3 g/un bol per os adulte
de 4 g
4. Romavermectin Avermectin Soluie 0,1 ml/5 kg g.v. trematode
B1 plus (prep.n 1% + injectabil, adulte
omologare) Clorsulon 10% s.c.
5. Fascioxamid Rafoxanid Boluri, 1 bol/20 kg g.v. Fasciole
per os tinere mai
mari de 6
spt.
6. Levamisol Levamisol Soluie 0,25 ml/10 kg Imuno-
7,5 % injectabil, s.c. g.v., 3 zile stimulator

REZULTATE I DISCUII

Rezultatele obinute sunt prezentate n tabelul 2.


Preparatul Fasciocid (s.a. Triclabendazol), administrat sub form de
soluie buvabil, a determinat o reducere marcant a gradului de anemie, cu
revenirea parametrilor eritrocitari n limite normale (valori medii): E de la 5,94 la
9,64 x 106/l, Hb de la 8,6 la 11,8 g/dl, iar Ht de la 28 la 32%.

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Se observ totodat un puternic efect leucopenic (leucocitele totale se reduc


de la 8,43 la 3,40 x 103/l) i trombocitopenic (trombocitele se reduc de la 3,6 la
1,1 x 105/l).
Aciunea exercitat de acest medicament asupra oulor de parazit, asupra
fasciolelor tinere i adulte, i asigur eficacitatea (9).
Preparatul Ovizol 300 (s.a. Albendazol), administrat oral sub form de
boluri, 0,3 g Albendazol/bol, a fost cel mai eficient dintre preparatele
administrate, determinnd restabilirea complet a numrului de E (de la 6,43 la
9,94 x 106/l), a Hb (de la 8,8 la 14 g/dl) i a Ht (de la 27 la 39 %); leucocitemia
s-a diminuat, reducndu-se de la 11,20 la 7,50 x 103/l (fiind n limite normale),
iar trombocitemia a cobort sub limita fiziologic inferioar (de la 2,70 la 0,80 x
105/l) (4).
Dei acioneaz doar asupra formelor adulte de Fasciola hepatica, acest
preparat devine eficient prin eliberarea i absorbia treptat a substanei active, cu
distrugerea lent a paraziilor pe msura migrrii n canaliculele biliare, nainte ca
noi forme adulte s devin hematofage.
Preparatul Helmizol 2,5 (s.a. Albendazol), suspensie buvabil 2,5 %,
administrat per os, nu a avut un efect evident asupra eritronului, hematocritul
meninndu-se sczut (28 %), dei E i Hb au nregistrat o uoar cretere (de la
6,43 la 7,54 x 106/l, respectiv de la 9,2 la 10,2 g/dl). Leucocitele totale din
sngele periferic au diminuat uor de la 9,05 la 8,45 x 103/l, iar trombocitele s-au
redus remarcabil, de la 6,7 la 2,5 x 105/l.
Eficiena acestui preparat este redus comprativ cu a preparatului Ovizol
300, dei conine aceeai substan activ.
Preparatul Romavermectin B1 plus, o combinaie ntre un antibiotic cu
aciune asupra nematodelor i artropodelor, i clorsulon, o sulfamid cu aciune
asupra trematodelor adulte, este eficient asupra endo- i exoparazitozelor.
Administrat lotului de ovine cu fascioloz, acest preparat a exercitat un efect
asupra eritronului, diminund uor gradul de anemie (E au crescut de la 5,80 la
8,47 x 106/l, Hb de la 8,6 la 12,2 g/dl, iar Ht de la 28 la 30 %). Totodat s-a
observat o diminuare a numrului de leucocite circulante de la 10,3 la 6,6 x 103/l
i a numrului de trombocite de la 3,5 la 2,1 x 105/l.
Preparatul Fascioxamid (s.a. Rafoxanid), administrat per os, sub form
de boluri, n-a determinat reducerea gradului de anemie; dimpotriv, boala a
evoluat, accentundu-se starea de anemie (valori mai mici pentru E: 6,23,
respectiv 6,17 x 106/l, Hb: 10,3, respectiv 9,3 g/dl i Ht : 28, respectiv 25 %) i
meninndu-se leucocitoza (nr. de L peste media normal a speciei : nainte de
tratament 9,48 x 103/l, dup tratament 9,05 x 103/l). Valoarea medie normal
pentru ovine adulte este de 8 x 103/l (Jain, 1986). De asemenea, s-a constatat o
uoar cretere a numrului de trombocite de la 3,3 la 5,1 x 105/l (rmnnd ns
n limitele normale ale speciei 2,5-7,5 x 105/l) (4).

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Dei acest preparat, derivat al salicil-anilidei, este cunoscut ca eficient


(96%) fa de trematodele tinere de peste 6 sptmni (9), efectul nu a fost
observat la oile luate n studiu, probabil datorit aciunii reduse asupra fasciolelor
adulte hematofage, deja ajunse n canaliculele biliare.
Preparatul Levamisol (s.a. Levamisol, izomerul levogir al Tetramisolului),
dei nu este trematodocid, s-a administrat unui lot de ovine cu fascioloz, avnd
n vedere efectul su imunomodulator. Intr-adevr, Levamisolul nu a redus starea
de anemie (E, Hb, Ht meninndu-se sczute : 6,52 x 106/l, 9,4 g/dl, Ht 27 %) ;
s-a constatat ns o diminuare a numrului de leucocite (de la 9,85 la 6,25 x
103/l) i absena modificrii numrului de trombocite (3,4 x 105/l nainte de
terapie, 3,2 x 105/l dup terapie).
Este cunoscut faptul c Levamisolul restabilete funcionalitatea celulelor
imunocompetente (polimorfonucleare, macrofage, limfocite) (1).
Lotul de ovine cu fascioloz la care nu s-a administrat nici un medicament
antiparazitar, a evideniat o accentuare a gradului de anemie prin diminuarea
valorilor parametrilor eritrocitari (nr. de E a continuat s scad de la 6,60 la 5,02 x
106/l, Hb de la 9,6 la 8,6 g/dl, iar Ht de la 30 la 26 %) i meninerea numrului
de leucocite ctre limita superioar a speciei (10,3 x 103/l la prima examinare i
11,0 x 103/l la a doua examinare, dup 30 zile de evoluie a bolii). Numrul de
trombocite a rmas nemodificat (3,50, respectiv 3,20 x 105/l) (4).

CONCLUZII

1. La ovinele infestate cu Fasciola hepatica, substanele benzimidazolice:


Ovizol 300 (avnd ca substan activ Albendazolul) i Fasciocid (preparat n
omologare, avnd ca substan activ Triclabendazolul) au aciune evident de
restabilire a valorilor normale ale eritronului (E, Hb, Ht) i au ca efect secundar
trombocitopenia.
2. Preparatul Helmizol 2,5, administrat per os, ca soluie buvabil, dei are
aceeai substan activ, nu are aceeai eficien ca Ovizol 300, administrat per os
sub form de boluri.
3. Romavermectin B1 plus (s.a Clorsulon - sulfamid) a determinat o uoar
reducere a gradului de anemie la oile cu fascioloz, precum i o diminuare a
leucocitemiei.
4. Preparatele Fascioxamid (s.a Rafoxanid derivat al salicilanilidei) i
Levamisol (s.a. izomerul levogir al tetramisolului derivat imidazo-tiazolic)
administrate ovinelor cu fascioloz au meninut starea de anemie i chiar au
accentuat gradul de anemie al animalelor bolnave (E, Hb, Ht au continuat s
scad).
5. Aciunea imunostimulatoare a levamisolului s-a evideniat prin
diminuarea numrului leucocite totale, rezultat al accenturii marginaiei i a
diapedezei leucocitare, cu infiltrarea esutului hepatic traumatizat.

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BIBLIOGRAFIE

1. Dobrescu, D., 1989 Farmacoterapie practic, vol.1, Ed. Medical, Bucureti.


2. Dulceanu, N., Terinte Cristina, Mitrea I.L., Polcovnicu Carmen, 2000 - Dicionar
Enciclopedic de Parazitologie, Ed. Academiei Romane, Bucuresti.
3. Iacob, Olimpia, 2002 Diagnosticul bolilor parazitare la animale, Ed. Ion Ionescu de
la Brad, Iai.
4. Jain, N.C., 1986 - Schalm's Veterinary Hematology. Lea-Febiger, Philadelphia.
5. Ingeborg, B., 1985 - Farmacodinamie pentru medici veterinari, Ed. Ceres, Bucuresti.
6. Kerr, M.G., 1989 - Veterinary Laboratory Medicine. Clinical Biochemistry and
Haematology. Blackwell Scientific Publication, Oxford.
7. Kondi, V., 1981 - Laborator clinic de hematologie. Ed. Medical, Bucureti.
8. Manolescu, N. i colab., 1999 Tratat de hematologie animal, vol I, Ed. Fundaiei
Romnia de mine, Bucureti.
9. Mitrea, I.L. 2002 Boli parazitare la animale, Ed Ceres, Bucureti.
10. Parvu Gh., 1992 Supravegherea nutriional-metabolic a animalelor, Ed.Ceres,
Bucureti.
11. Suteu, I., Varctic N., Cozma V., 1996 - Diagnosticul i tratamentul parazitozelor la
animale, Ed. Ceres, Bucuresti.

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OBSERVAII PRIVIND EFECTUL UNOR SUBSTANE


ANTIPARAZITARE ASUPRA ENZIMELOR SERICE DE
ORIGINE HEPATIC LA OVINE CU FASCIOLOZ
ON THE EFFECT OF SOME ANTIPARASITIC SUBSTANCES ON THE SERUM
ENZYMES OF HEPATICAL ORIGIN IN OVINES WITH FASCIOLIASIS

E. PURICE
C.S.V. Vidra, jud.Vrancea
P. CURA, Geta PAVEL, V. BOGHIAN
U.S.A.M.V. Iai

Different antiparasitic drugs were administrated to 6 flocks of ovines


with fascioliasis, each flock being made of 5 sheeps. A flock has been kept
as a non-treated witness (Fasciocid; Ovizol 300; Helmizol 2,5;
Romavermectin B1 plus; Fascioxamid; Levamisol). Before and after 30 days
from drug administration Serum transaminase (AST, ALT and GGT) and
alkaline phosphatase (ALP) were measured in all investigated animals.
Before the application of the therapy with various antiparasitic
substances, high averaged values for AST and ALP and averaged values
between the physiological limits for ALT and GGT have been measured in
ovines with fascioliasis. After the antiparasitic substance administration, in
most of the cases AST (which reflects the hepato-cellular dysfunction), kept
staying at a high value, over the superior limit of reference, although by
comparation with its initial values, it oscillates. The seric level of ALP
(indicator of some moderate destruction of the liver parenchyma, but
mainly of the hepato-biliary dysfunction) kept growing after the therapy
with the following antiparasitic substances: Fasciocid, Ovizol 300,
Helmizol, Romavermectin and Levamisol. An exception makes the therapy
with Fascioxamid which determines the return of the ALP level at its
physiological limit. ALT increased over the initial value, sometimes over the
physiological superior limit, and GGT (chronicity indicator of the hepato-
cellular damage) kept staying within the physiological limits or dropped
bellow the inferior limit of reference (for the therapy with Fasciocid and
Levamisol).

Key words: fascioliasis, sheeps, benzimidazolic therapy, AST, ALT, GGT, ALP

In fascioloza ovin, n faza de stare a bolii, examenul paraclinic evideniaz


o serie de modificri ale profilului biochimic hepatic: creterea transaminazelor,
creterea fosfatazei alcaline i scderea proteinelor serice. Aceste modificri se
datoreaz distruciilor masive ale parenchimului hepatic i ale canaliculelor biliare
(1, 3, 6).

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In prezenta lucrare se urmrete efectul substanelor antiparazitare asupra


transaminazelor serice i asupra fosfatazei alcaline, n condiiile n care acestea
prezint modificri importante n cursul evoluiei fasciolozei.

MATERIAL I METOD

Cercetrile s-au efectuat ntr-un efectiv de ovine cu fascioloz


(diagnosticat clinic i paraclinic). De la 35 de animale grupate pe loturi de cte 5
animale s-au recoltat probe de snge nainte i dup aplicarea terapiei cu diverse
substane antiparazitare. Din probele de snge s-au determinat enzimele: AST
(aspartat aminotransferaza), ALT (alanin aminotransferaza), PA (fosfataza
alcalin) i GGT (gama-glutamiltransferaza), utiliznd analizatorul semiautomat
EOS 880 Plus i chituri specifice.
Lotul 1 a fost tratat cu Fasciocid (s.a. Triclabendazol 10%), administrat sub
form de soluie buvabil, 5 ml/50 kg g.v. Lotul 2 a fost tratat cu Ovizol 300
(Albendazol 0,3mg/bol de 4 g), administrat per os, 2 boluri/animal. Lotul 3 s-a
tratat cu Helmizol 2,5 (s.a. Albendazol 2,5%), administrat per os, 3 ml/10 kg g.v.
Lotul 4 s-a tratat cu Romavermectin B1 plus (s.a. avermectin 1% i clorsulon
10%), soluie injectabil, administrat s.c., 0,1 ml/5 kg g.v. Lotul 5 a fost tratat cu
Fascioxamid (s.a. Rafoxanid), administrat per os, un bol/ 20 kg g.v. Lotul 6 s-a
tratat cu Levamisol 7,5%, administrat sub form de soluie injectabil, s.c., 0,25
ml/10 kg g.v., 3 zile (4, 6, 7). Lotul 7 nu a fost supus terapiei antiparazitare i a
constituit lotul martor.

REZULTATE I DISCUII

Rezultatele obinute sunt prezentate n tabelul 1.


Se remarc faptul c la toate animalele supuse investigaiilor, inainte de
administrarea medicaiei antiparazitare, enzimele serice AST i PA au valori
crescute peste limita superioar a speciei.
AST aspartat aminotransferaza - este o enzim ubicvitar, dar se gsete
predominant n ficat, muchi i eritrocite. La oile cu fascioloz, migraia
fasciolelor tinere prin parenchimul hepatic determin distrucii masive celulare,
astfel c enzimele se elibereaz n cantitate mai mare n snge (2, 5).
PA fosfataza alcalin este, de asemenea, o enzim larg rspndit n
organism, dar se gsete n special n osteoblaste, ficat i peretele intestinului
subire. Dup definitivarea creterii, originea acestei enzime serice rmne
predominant hepatic. PA seric este un indice al funciei hepato-biliare. Bolile
parenchimului hepatic determin creteri moderate ale PA, pe cnd cele ale
canaliculelor biliare, cauzeaz creteri accentuate ale acestei enzime serice (2, 5).
La loturile de animale cu fascioloz luate n studiu, valorile medii pentru
AST au avut creteri moderate sau accentuate (ntre 193,7 U/l i 292 U/l) fa de
valorile de referin (49-123 U/l), n timp ce valorile medii pentru PA au prezentat
i valori n limitele fiziologice (102,1 U/l), precum i creteri (intre 182,3 i 302,4
U/l) fa de valorile de referin (26,9-156,1 U/l) (8).

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In fascioloz, creterea PA se explic prin aciunea mecanic-iritativ a


fasciolelor adulte din canalele biliare, care determin imflamaie cronic,
colangit i ciroz pericanalicular; se poate produce i obstruarea canaliculelor
biliare de dimensiuni mici, creterile PA fiind drastice n acest caz (6).
ALT i GGT, nainte de terapie, la oile cu fascioloz investigate, au
prezentat valori n limitele fiziologice. ALT a prezentat valori medii ntre 32,4 i
43,2 U/l, ctre limita superioar a valorilor de referin ale speciei (14,8-43,8 U/l).
GGT a avut valori medii ntre 19,8 i 38,3 U/l, extrem de variabile, dar
ncadrndu-se n limitele de referin (19,6-44,1 U/l) (8).
ALT, dei este considerat fr semnificaie la erbivore (fiind de mare
valoare diagnostic n afeciunile hepatocelulare la om i carnivore), ea a fost
determinat cu scopul evidenierii efectului substanelor antiparazitare (2).
GGT are semnificaie clinic pentru bolile hepatice, inclusiv la erbivore,
nregistrnd creteri ntr-o faz mai avansat a bolii, cnd celelalte enzime ncep
s scad (2, 5).
Tabelul 1
Valorile medii ale enzimelor serice la ovinele tratate cu substane
antiparazitare fascilocide

Preparatul Momentul Parametrii biochimici investigai


Lot de
antiparazitar exam. AST ALT PA GGT
animale
administrat biochimic U/l U/l U/l U/l
I 258,2 33,6 302,4 19,8
Lot 1 (n=5) Fasciocid
II 179,4 41,9 327,5 10,1
I 215,0 32,4 102,1 29,3
Lot 2 (n=5) Ovizol 300
II 183,3 42,6 222,4 35,4
I 292,3 32,6 233,4 38,3
Lot 3 (n=5) Helmizol 2,5
II 442,8 46,4 312,7 40,9
Romavermectin I 218,3 43,2 189,4 33,3
Lot 4 (n=5)
B1 plus II 225,4 52,4 224,5 24,4
I 276,7 33,5 182,3 26,9
Lot 5 (n=5) Fascioxamid
II 254,0 45,7 126,7 30,2
I 260,3 38,5 281,2 23,6
Lot 6 (n=5) Levamisol
II 258,9 45,6 316,3 18,4
Lot 7 (n=5) I 193,7 41,9 231,0 31,3
-
II 195,7 44,8 107,3 20,5
Valori de referin (Merck, 1998) 49-123 14,8-43,8 26,9-156,1 19,6-44,1
I - Inainte de tratam; II La 30 zile dup tratament

In urma terapiei cu Fasciocid, AST se reduce dar nu revine n limite


fiziologice, PA continu s creasc uor, ALT crete, dar rmne n limitele
fiziologice, iar GGT se reduce sub limitele fiziologice (fig.1a).
Dup terapia cu Ovizol 300, de asemenea, se reduce AST fr a reveni n
limitele fiziologice, iar PA crete de la valori fiziologice peste limita superioar a
speciei; att ALT, ct i GGT nregistreaz creteri reduse fr a depi limitele
fiziologice de referin (fig.1b).

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Dup 30 zile de la administrarea preparatului Helmizol 2,5, AST i PA


continu s creasc mult peste limita superioar a speciei, uoare creteri
nregistrnd i ALT i GGT (GGT nedepind limita superioar de referin)
(fig.1c).

Nivelul seric al enzim elor hepatice la ovine tratate Nivelul seric al enzim elor hepatice dupa terapia cu
cu Fasciocid Ovizol 300

U/l U/l
350 250
300
200
250
Inainte de 150 Inainte de
200
tratament tratament
150 100
Dupa 30 zile Dupa 30 zile
100
50
50
0 0
AST ALT PA GGT AST ALT PA GGT

a b
Nivelul seric al enzim elor hepatice dupa terapia cu Nivelul seric al enzim elor hepatice dupa terapia cu
Helm izol Rom averm ectin B1 plus

U/l U/l
500 250

400 200

300 Inainte de 150 Inainte de


tratament tratament
200 Dupa 30 zile 100 Dupa 30 zile
100 50

0 0
AST ALT PA GGT AST ALT PA GGT

c d
Nivelul seric al enzim elor hepatice dupa terapia cu Nivelul seric al enzim elor hepatice dupa terapia cu
Fascioxam id Levam isol

U/l U/l
300 350
250 300
250
200 Inainte de Inainte de
200
150 tratament tratament
150
100 Dupa 30 zile Dupa 30 zile
100
50 50
0 0
AST ALT PA GGT AST ALT PA GGT

e f
Fig.1. Modificarea nivelului seric al enzimelor hepatice dup terapia cu substane
antiparazitare fasciolocide.

Dup tratamentul cu Romavermectin B1 plus, AST s-a meninut la un nivel


ridicat, ca i PA, peste limitele fiziologice superioare, ALT a crescut de la valori
normale peste limita superioar a speciei, iar GGT a sczut uor rmnnd ns n
limitele fiziologice de referin (fig.1d).

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Dup administrarea preparatului Fascioxamid, s-a nregistrat o uoar


reducere a AST, cu meninerea valorii medii peste limita superioar, precum i
reducerea Pa la valori medii care se ncadreaz n limitele fiziologice de referin.
ALT a crescut, depind limitele fiziologice, iar GGT a crescut uor fr a depi
limita superioar a speciei (fig.1e).
In urma terapiei cu Levamisol, AST s-a meninut la un nivel ridicat peste
limita superioar, PA a continuat s creasc, ALT a crescut, de asemenea, peste
limita superioar de referin, iar GGT a sczut uor sub limita inferioar a speciei
(fig.1f).
Oile cu fascioloz din lotul martor, care n-au fost tratate cu medicaie
antiparazitar, au prezentat o valoare crescut a AST, cu meninerea aceleiai
valori pe parcursul a 30 zile, o uoar cretere a ALT, o uoar reducere a GGT i
o scdere evident a fosfatazei alcaline, cu revenirea n limitele fiziologice (fig.2).
AST, ca i n cazul lotului martor netratat, la cele ase loturi supuse
tratamentului cu substane antiparazitare fasciolocide, se menine la un nivel seric
ridicat, peste limita superioar fiziologic , dei s-au nregistrat i valori mai
sczute, sau creteri accentuate fa de valoarea iniial. Astfel, preparatele
Fasciocid i Ovizol 300 se dovedesc mai eficiente n privina diminurii
distruciilor celulare hepatice, n timp ce Helmizol 2,5, dimpotriv a accentuat
distruciile tisulare.

Nivelul seric al enzim elor hepatice la ovine


cu fascioloza - lot m artor netratat

U/l
250

200

150
Prima investig.
100 Dupa 30 zile
50 Fig.2. Modificarea nivelului seric al
0 enzimelor hepatice la oile cu
AST ALT PA GGT fascioloz nesupuse terapiei cu
medicaie antiparazitar

ALT, avnd iniial valori n limite fiziologice, att la lotul martor netratat,
ct i la loturile supuse terapiei antiparazitare, dup 30 zile, a nregistrat uoare
creteri n limitele fiziologice sau peste acestea, aspect ce se coreleaz cu
modificarea AST.
PA a nregistrat creteri i meninerea peste valorile normale de referin la
toate loturile supuse investigaiei, excepie fcnd lotul martor i lotul tratat cu
Fascioxamid, la care fosfataza alcalin a sczut la valori care se ncadreaz n
limitele fiziologice ale speciei. In acest caz, s-ar putea presupune c Fascioxamid
care distruge formele tinere de Fasciola hepatica aflate n migraie, determin o
diminuare a procesului inflamator hepatic, astfel c nivelul seric al PA se
restabilete. In acelai mod, procesul patologic celular, chiar n absena terapiei,

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urmeaz un proces rezolutiv la nivelul parenchimului hepatic, iniial distrus, i


abia ntr-un stadiu mai avansat al bolii, probabil peste mai mult de 30 zile,
fasciolele adulte produc colestaz, colangit i, ca urmare o nou cretere a PA.
GGT nu a nregistrat la nici unul dintre loturile investigate creteri peste
limitele fiziologice, ceea ce arat c evoluia procesului patologic nu a fost
suficient de ndelungat pentru a determina creterea nivelului seric al acestei
enzime.

CONCLUZII

1. Inainte de aplicarea terapiei cu diverse substane antiparazitare, ovinele


cu fascioloz au nregistrat valori medii crescute pentru AST i PA i valori medii
n limitele fiziologice de referin pentru ALT i GGT.
2. Dup administrarea substanelor antiparazitare diferite la cele 6 loturi de
ovine investigate, AST i PA, n majoritatea cazurilor, rmn la valori ridicate,
peste limitele superioare de referin, dei fa de valorile iniiale se nregistreaz
unele oscilaii (n sensul creterii sau scderii).
3. Creterea nivelului seric al PA se accentueaz dup terapia cu substane
antiparazitare: Fasciocid, Ovizol 300, Helmizol, Romavermectin i Levamisol;
face excepie terapia cu Fascioxamid, care determin revenirea nivelului PA n
limite fiziologice, ca i n cazul lotului martor netratat.
4. Sub aciunea substanelor antiparazitare, dup 30 de zile de la
administrare, ALT nregistreaz o cretere fa de valoarea iniial, uneori peste
limita superioar fiziologic, iar GGT rmne n limite fiziologice sau
nregistreaz o scdere sub limita inferioar de referin (n cazul terapiei cu
Fasciocid i Levamisol).
5. Modificrile variabile observate dovedesc faptul c preparatele
antiparazitare utilizate n experiment au eficien n terapia fasciolozei, dar
prezint i un anumit grad de hepatotoxicitate.

BIBLIOGRAFIE

1. Dulceanu, N., Terinte Cristina, Mitrea I.L., Polcovnicu Carmen, 2000 - Dicionar
Enciclopedic de Parazitologie, Ed. Academiei Romane, Bucuresti.
2. Ghergariu, S., AL. Pop, L. Kadar, Mariana Spnu, 2000 Manual de laborator
clinic veterinar, Ed.All, Bucureti.
3. Iacob, Olimpia, 2002 Diagnosticul bolilor parazitare la animale, Ed. Ion Ionescu de
la Brad, Iai.
4. Ingeborg B., 1985 - Farmacodinamie pentru medici veterinari, Ed. Ceres, Bucuresti.
5. Kerr, M.G., 1989 - Veterinary Laboratory Medicine. Clinical Biochemistry and
Haematology. Blackwell Scientific Publication, Oxford.
6. Mitrea I.L., 2002 Boli parazitare la animale, Ed Ceres, Bucureti.
7. Suteu, I., Varctic N., Cozma V., 1996 - Diagnosticul i tratamentul parazitozelor la
animale, Ed. Ceres, Bucuresti.
8. ***, 1998 - Merck Veterinary Manual, Eight Edition, Merck-Co., Inc. Whitehouse
Station, N.J., USA.

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CASTRATION OF DOG BY SCROTAL SEPTUM ECTOMY


S. CIOBANU, I. BURTAN, M. FNTNARU,
L.C. BURTAN, Roxana TOPAL
U.S.A.M.V. Iai

Castration (orchiectomy) is one of the more frequently performed


surgical procedures in veterinary practice.
The male gonads ablation is the main purpose and can be done in
many ways. One of this is the ectomy of scrotal septum and partial ectomy
af scrotum.
The first step is to identify and distinguish the scrotal septum with
two hemostatic forceps placed anterior and posterior. Vertical traction is
then applied and the scrotum forms a wrincle that will be exciset with
surgical scissor. By this procedure the testis are exteriorizated and operator
is able tu performe an open or close castration. The surgical wound is
sutured with a simple continous or interrupted suture.

Castration of the dog is a routine intervention with many indications.


Although the orhidectomy tehnique is well known the operation has postoperative
complications. Those postoperative complications depends on applied method and
acces way that we chose for testis ablation.
In this paper work we presented the tehnique that involves the ectomy of
scrotal septum. This acces way has many advantages and can be applied on
heathy dogs and on dogs with testis and scrotal diseases.

MATERIAL AND METHOD

The researche was perfomed on 30 dogs that were castrated by this


tehnique in Surgycal Clinic of Veterinary Medicine Faculty.
Equipment: Standard general surgical pack and suture. For general
anesthesia we used xylasin 2% and ketamin 10% in proper doses, and for local
anesthesia we used xilin 2% infiltrated in scrotum.
The animal is placed in dorsal recumbency and prescrotal area is
aseptically prepared.
Operatory tehnique:
- Identify and distinguish the scrotal septum with two hemostatic forceps
placed anterior and posterior (foto 1).
- By excision of the wrinkle that formed under those two hemostatic forceps
we executated the ectomy of scrotal septum (foto 2). The wrinkle is formed by a
part of scrotum and scrotal septum. If the hemostatic forceps are placed in a large
distance one from another, the wrinkle will be bigger. To avoid tension of the
suture we have to leave enough skin after excision. After the extomy the testis are
exteriorizated and we can proced an close castration (foto 3). We assured

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hemostasis using transfixation ligature on the spermatic cord, with absorbable


suture material (foto 4, 5).
- Surgical wound is sutured by using a two-layer suture (foto 6). The first
suture it was a simple interrupted suture with absorbable material and the second
was an intradermic simple continuous suture.

Foto 1. Identify and distinguish the scrotal Foto 2. Ectomy of scrotal septum
septum

Foto 3. Exteriorization of testis Foto 4. Ligature on the spermatic cord and


testis ablation

Foto 5. Aspect of surgical wound afer the Foto 6. Final aspect of surgical wound
first layer suture

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RESULTS AND DISCUSSIONS

By ectomy of scrotal septum and partial ectomy of scrotum we remove the


unnecesary tissue. To avoid tension of the suture we have to leave enough skin
after excision. The purpose of the two-layer suture is to draw the skin near the
subcutaneous tissue, avoiding exsudat apparition.
We recomand intredermic suture to prevent licking or chewing the suture.
The ectomy tehnique is relative simple and can be performed on heathy
dogs and on dogs with testis and scrotal diseases.

CONCLUSIONS

We evaluated a new surgical acces way for orhiectomy at dogs


This new acces way by ectomy of scrotal septum is easy to perform, faster
and more secure avoiding the risks of postoperative complications
Comparing with other tehniques, this one can be performed in any situation
that requires orhiectomy.

BIBLIOGRAPHY

1. Birchard S., Sherding R. Small Animal Practice, W.B. Saunders Co., Philadelphia
2. Bojrab M.J., Birchard S., Tomlinson J., 1990 Current Tehniques in Small Animal
Surgery, Lea and Febiger, Philadelphia
3. Burtan I., 1991 Curs de propedeutica si tehnica chirurgicala. C.M. Inst.Agr. Iasi
4. Cristea I. si colab., 1983 Propedeutica chirurgicala si tehnica operatorie
veterinara. Ed. did. si pedag. Bucuresti
5. Gourley I.M., Clare Gregory, 1992 Small Animal Surgery, Gower Medical
Publishing, New York
6. Grigorescu I., 1973 Propedeutica si medicina operatoare. Ed. did. si pedag.
Bucuresti
7. Igna C., 2005 Tehnici chirurgicale veterinare. Ed. Brumar.
8. Mialot J.P. Pathologie de la reproduction chez les carnivores domestiques. Ed. du
Point Veterinaire.

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POSIBILITI DE PREVENIRE
N NEOPLAZIA MAMAR LA CANINE
PREVENTION POSIBILITIES IN MAMMARY NEOPLASIA AT CANINES

L.C. BURTAN, I. BURTAN,


M. FNTNARU, S. CIOBANU,
Roxana TOPAL
U.S.A.M.V. Iai

The prevention of mammary tumours is easier than the treatment,


therefor we identified some posibilities to reduce or to supress mammary
cells modifications.
Mammary cells are able to multiplicate uncontrolled in cases of
mammary traumatisms, chemical control of heat cycle, unwanted
pregnancy supress and pseudopregnancy lactation.
An suitable treatment of mammary open traumatisms and
ovariectomy are some posibilities to prevent mammary cells revolution.

Key words: canine, mammary cancer, cicatrization, unwanted pregnancy,


pseudopregnancy lactation, chemical control of heat cycle, ovariectomy

Dei profilaxia neoplaziei mamare este o himer totui exist suficiente


momente n viaa unei femele, care, dac sunt evaluate i tratate cu atenie pot
preveni o cancerogenez mamar. n acest sens amintim traumatismele mamare i
revoluiile celulare de la acest nivel (2, 5, 10).
Revoluiile celulare mamare, reprezentate de transformrile care au loc la
acest nivel, se pot ntlni n ntreruperea gestaiei nedorite, n lactaia de
pseudogestaie i n controlul chimic al estrului (1, 3, 4, 6, 8, 9). De asemenea,
ntreruperea brusc a lactaiei, imediat dup parturiie, duce la modificri celulare
mamare (5, 7).
Pentru prevenirea acestor revoluii mamare s-a utilizat controlul chirurgical
al reproduciei prin ovarectomie sau ovareohisterectomie (7, 8, 10).

MATERIAL I METOD

Materialul de studiu a fost reprezentat de cazuistica oferit de Clinica


Chirurgical a Facultii de Medicin Veterinar Iai i Cabinete medicale
veterinare private.
Traumatismele mameloanelor sau esutului mamar, care creeaz soluii de
continuitate, au fost urmrite pentru a putea concluziona asupra apariiei
momentului n care, o celul capabil s se multiplice pentru regenerare deviaz
de la activitatea programat. n acest sens au fost urmrite 15 femele cu plgi
accidentale n regiunea mamar. Plgile nu au fost supuse prelucrrii chirurgicale

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i suturii fiind lsate prad instinctelor terapeutice ale animalului, respectiv


lingerii care irit continuu esutul de reparaie.
ntruct glanda mamar se afl sub controlul secvenial al steroizilor
ovarieni, investigaiile au fost orientate spre stabilirea unor cauze capabile a
produce revoluii celulare n acest esut. n acest sens, au fost luate n studiu 276
femele necastrate, la care, pe baza datelor anamnetice au primit medicaie pentru
ntreruperea gestaiei nedorite, pentru combaterea lactaiei de pseudogestaie
sau pentru controlul chimic al estrului, evenimente care s-au repercutat asupra
esutului mamar.

REZULTATE I DISCUII

Procesul de reparaie tisular, la cele 15 cazuri, s-a desfurat ntr-un timp


relativ lung, evolund spre vindecare prin nmugurire, spre cicatrizare excesiv,
lipsa cicatrizrii sau spre cancerizare, datele fiind prezentate n tabelul 1.
Tabelul 1
Date centralizate privind influena iritaiilor asupra cicatrizrii
plgilor mamare la cea

Rezultatele tratamentului instinctiv


Nr. de plgi Vindecare Plag aton Cheloidizare Tumorizare
Nr. % Nr. % Nr. % Nr. %
15 7 46,7 2 13,3 3 20 3 20

Analiznd datele din tabelul 1 constatm gravitatea lipsei de intervenie


terapeutic n plgile mamare. Iritaiile produse de animal prin toaletarea plgii, la
care se adaug cele determinate de factorii de mediu, tulbur evoluia normal a
procesului de nmugurire, nct numai 46,7% din cazuri nregistreaz o evoluie
normal. Totusi, trebuie s menionm c reparaia s-a produs dup 25-35 de zile.
Dei timpul de vindecare trebuie corelat cu mrimea plgii, putem aprecia c
acesta a fost suficient de mare, comparativ cu cel realizat n practic prin
prelucrare chirurgical i sutur primar care este de 10-12 zile.
La dou cazuri, respectiv 13,3% traumatizarea prin lingere a esutului de
nmugurire a determinat aplatizarea acestuia i lipsa de reacie perifocal,
transformnd plaga ntr-un esut aton. n acest caz s-a intervenit prin extirparea
esutului aton i sutur primar.
Iritaiile produse prin lingere pot determina o tulburare a prezenei celulare
implicate n reparaia tisular.
Ruperea echilibrului ntre factorii care stimuleaz vindecarea i cei care o
inhib, conform reparaiei programate, face ca esutul de nmugurire s creasc n
exces. n final este influenat epidermizarea ducnd la creterea n volum a
esutului de reparaie care se transform ntr-o pseudotumor, respectiv cheloid,
diagnosticat la 20% din cazuri.

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La 3 cazuri, respectiv 20% din femelele urmrite, procesul hiperplazic


cutanat a interesat i esutul mamar care a crescut n exces. Baza mamelonului
avea dimensiunile unei prune, cu consisten crescut, iar pielea cptase aspectul
unei cicatrici hipertrofice. Din anamnez a rezultat c plgile accidentale aveau
vechime de 65-75 zile.
n ceea ce privete rolul modificrilor celulare, din glanda mamar, pentru
declanarea tumorigenezei, datele obinute sunt centralizate n tabelul 2.
Tabelul 2
Corelarea prevalenei tumorilor mamare
cu tulburri ale ciclului sexual la cea

Femele cu tulburri ale ciclului sexual


Nr. Femele cu Din care
femele ciclul Controlul
cu sexual Total Gestaie Lactaie de chimic
tumori normal nedorit pseudogestaie intermitent al
mamare estrului
Nr % Nr % Nr % Nr % Nr %
276 106 38,4 170 61,6 25 14,7 114 67 31 18,3

Analiza detelor nscrise n tabelul 2 evideniaz faptul c ocurile


hormonale aprute n viaa unei femele pot sta la baza neoplaziei mamare.
Glanda mamar, ca organ hormono-dependent, suport aceste ocuri prin
modificri n populaia celular.
Astfel, din cele 276 de femele cu tumori mamare, 38,4% au prezentat anual
o evoluie normal a ciclului sexual, iar 61,6% au inregistrat diferite tulburri ale
ciclului sexual, impunnd administrarea de produse hormonale pentru reglare.
Din cele care au prezentat tulburri 14,7% au primit tratament hormonal
(Sintofolin, Mesalin) pentru a evita instalarea gestaiei nedorite, iar 18,3% au fost
supuse tratamentului hormonal (Medroxiprogesteron) pentru controlul chimic al
estrului.
De asemenea, 67% din femelele cu tulburri ale ciclului sexual au primit
tratament hormonal pentru combaterea lactaiei de pseudogestaie. Tratamentul
hormonal a fost local (Mastoprofen, Progestagel) i general (Covinan,
Bromocriptina), medicaie capabil s determine inhibarea secreiei de prolactin,
reducnd astfel brusc activitatea acinilor glandulari.
Prezentarea acestor posibile cazuri ale tumorigenezei mamare i modul n
care pot fi combtute sunt argumente care conduc la concluzia c neoplazia
mamar poate fi o iatropatie. Neglijena n terapia plgilor regiunii mamare,
abuzul de hormoni pentru combaterea dereglrilor ciclului sexual, sau pentru
stoparea acestuia sunt momente care tulbur dinamica celular a glandei mamare,
expunnd la apariia celulei capabile s se multiplice anarhic. De asemenea,
rezultatele prezentate evideniaz rolul controlului chirurgical al reproduciei la
canine prin ovarectomie.

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Planningul canin prin ovarectomie este o metod tiinific i sigur, care


contribuie la prevenirea tumorilor mamare. n acest sens am efectuat un studiu pe
123 femele care au fost supuse ovarectomiei la vrste diferite pentru a corela
riscul apariiei neoplaziei cu numrul ciclurilor sexuale, datele obinute fiind
prezentate n tabelul 3.
Tabel 3
Corelarea vrstei la castrare cu frecvena tumorilor mamare

Femele Vrsta n ani la castrare


castrate care Nr cazuri 1 2 3 4 5 6 7 8 9
au contractat 123 2 7 17 21 26 33 9 6 2
tumori % 1,6 5,7 13,8 17 21,1 27 7,3 4,9 1,6
mamare

Datele nscrise n tabelul 3 demonstreaz c ovarectomia executat la vrst


timpurie reduce posibilitatea apariiei tumorilor mamare. Astfel, 1,6% din
femelele cu tumori mamare au fost supuse ovarctomiei la vrsta de 1 an, deci
dup primul ciclu sexual, iar 5,7% au fost castrate la vrsta de 2 ani.
Pe msur ce femela nainteaz n vrst crete riscul apariiei tumorilor
mamare, curba fiind ascendent pn la vrsta de 7 ani cnd proprietarul accept
ovariectomia ca soluie de a elimina monta i lactaia de pseudogestaie. Femelele
castrate la 4-6 ani prezint cel mai mare risc de a face tumori mamare, cu 17%
castrate la 4 ani, 21,1% la 5 ani i 27% la 6 ani. Pn la aceast vrst femelele au
fost folosite la reproducie, iar la unele s-a ntrerupt ciclul sexual pe cale chimic.

CONCLUZII

Plgile mamare, dac nu sunt tratate corespunztor pot fi plasate n rndul


factorilor oncoinductori. Consecutiv acestor incidente 20% din plgi pot tumoriza
dup o perioad de 65-75 zile, iar 20% pot evolua spre cheloid cicatriceal care
este o pseudotumor.
Tulburrile ciclului sexual pot interveni n cancerogeneza mamar. Acestea
sunt reprezentate de gestaia nedorit (14,7%), lactaia de pseudogestaie (67%) i
controlul chimic intermitent al estrului.
Ovarectomia executat la vrst timpurie reduce considerabil riscul de a
face tumori mamare. Riscul crete de la 1,6% din femele cu tumori dac
ovarectomia s-a executat la 1 an, respectiv dup 1-2 cicluri sexuale.
Femele castrate dup 4-6 ani pot face tumori mamare n proporie de 21,1%
respectiv 27%.

BIBLIOGRAFIE

1. Avallone, L., Lombardi, P., Florio, S., 1994 Steroid receptors distribution in
canine mammary tumour. Acta Medica Veterinaria, 40, 2, 105-112.

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2. Baba, A.I.r 2002 - Oncologie comparat. Ed, Academiei Romne. Buc.


3. Baker, R.W., 1994 - Use of tamoxifen in the control of canine mammary neoplasia.
Vet. Record 134, 1-24.
4. Broiu, I.A., Seiciu, FL., 2004 Tratat de reproducie la animale. Ed. BIC ALL Buc.
p.254-258.
5. Burtan L.C., 2005 Contribuii la studiul oncopatiilor mamare la cine, Tez de
doctorat, U. S. A. M. V., Iai
6. Bonca, Gh., Cernescu, H., Ardelean, V., Mircu, C, Bedrosian, Ed., 1995 -
Cercetari comparative ale terapiei lactaiei nervoase la cea cu steroizi sexuali,
derivai ergolinici i revulsive locale. Rev. Rom. M.V., vol.5, nr.3, p.265-269.
7. Burtan, I., 2004 - Patologie chirurgical general veterinar. Ed. I. Ionescu de la
Brad Iai 122-131
8. Buff, S., 2001 Sterilisation tres precoce: de nombreux avantages. Le Point
Veterinaire, 221, 52-54.
9. Picavet Sophie, 1992 - Lactation de pseudogestation chez la chiene - Reproduction
du chien et du chat. P.C.M.A.C. 179-195.
10. Tnase, A., 1999 - Tratamentul complex chirurgical, chimio i imunoterapeutic al
cancerelor mamare la canidae i feline. Teza de doctorat U.S.A.M.V. Buc.

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REDUCEREA CHIRURGICAL N HERNIA PERINEAL


A VEZICII URINARE LA UN CINE PEKINEZ:
RAPORT DE CAZ
SURGICAL REDUCTION IN THE PERINEAL HERNIA OF URINARY BLADDER IN A
PEKINGESE DOG: CASE RAPORT

P. CAZACU, M. MARE
U.S.A.M.V. Iai

In this paper is presented a case of perineal hernia of urinary


bladder in a 11- year- old male dog, Pekingese that after anamnestic
investigations and clinical examination, it was diagnosed with perineal
hernia of urinary bladder and subsequently confirmed during the surgery.
We presented the mode of surgical resolution by using of trick in basic
techniques for reconstruction of pelvic diaphragma. The reconstruction of
serous hernial sac, through the torsion and ligature, like in techniques
applied from ombilical hernia and subsequently closure of muscular pelvic
diaphragma.

Key words: male dog, Peckingese, perineal hernia of urinary bladder,


peritoneal closure, muscular pelvic diaphragma closure.

Hernia perineal a vezicii urinare (H.P.V.U.) este o modificare topografic


caracterizat prin retroflexia vezicii urinare i hernierea acesteia, acoperit sau nu
de peritoneu, lateral anusului, prin penetrarea structurilor ce alctuiesc diafragma
pelvin (m. ridictor al anusului, m. coccigieni, m. sfincter anal extern, fascia
perineal).
H.P.V.U. este o afeciune chirurgical rar ntlnit n practica medical
veterinar, prevalena sa fiind cuprins ntre 0,1-0,4 % din cazurile prezentate n
clinicile veterinare (1). Rasele cel mai frecvent afectate, citate n literatur sunt:
Boxer, Boston Terrier, Pekinez, Teckel, Welsh Corgi, Collie i Ciobanesc englez
Bobtail. (1, 2, 6, 13, 14, 19). Totui, rase ca Boston Terrier, Peckinez i Boxer
prezint i un risc familial (12, 14).
H.P.V.U. afecteaz cel mai frecvent cinii masculi, necastrai, cu vrsta
cuprins ntre 7 i 9 ani (1, 6, 16).
n etiopatogenza H.P.V.U., sunt ncriminai mai muli factori: imbalan
hormonal, predispoziia congenital, slabirea structurilor diafragmei pelvine,
bolile prostatei i constipaia cronic (3, 6, 14, 16).
Expresia nalt a receptorilor pentru relaxin n musculatura diafragmei
pelvine sugereaz c aceasta joac un rol important n patogeneza herniei
perineale (H.P.) (15).

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Pentru c nici unul din aceti factori nu a fost demonstrat, originea acestor
tulburri este probabil o combinaie a acestora (3).
La cine, fundul de sac dorso-rectal al peritoneului parietal (excavaia
sacro-rectal), n regiunea pelvin, este foarte profund ajungnd aproape de
diafragma pelvin, foarte aproape de anus, aspect anatomic ce favorizeaz apariia
frecvent a herniilor perineale la aceast specie (10, 17).
Clinic, H.P.V.U. se caracterizeaz prin apariia unei tumefacii n regiunea
perineal. Aceasta poate fi uni- sau bilateral. Hernia perineal unilateral este
observat aproximativ la 2/3 din cazuri, iar 80% dintre acestea se gsesc pe partea
dreapt (2, 3, 6). H.P.V.U. se ntlnete cu o frecven de 29% din cazurile de
H.P. dup Brissot, iar Hosgood o ntlnete cu o frecven de 20%.
Organele pot hernia printre m. coccigien i m. ridictor al anusului sau
frecvent ntre m. sfincter anal extern, m. ridictor al anusului i m. obturator
intern (6, 12, 16).

MATERIAL I METOD

Subiectul este un cine mascul Peckinez, n vrst de 11 ani, criptorhid,


prezentat la consultaie cu un istoric de inapeten, polidipsie, vomituriie i voma
apei imediat dup consum, apariia unei tumefacii n regiunea perineal, anurie i
lipsa defecrii i nrutirea strii generale a animalului. Aceste simptome au
aprut i s-au accentuat nainte cu 24 ore de prezentarea animalului la
consultaie.
n momentul consultaiei, la examenul clinic, animalul prezenta depresie a
sistemului nervos central, ataxie i astazie, schimbndu-i frecvent decubitul
lateral stng cu cel drept. Reflexul palpebral este ntrziat, cinele prezint o
deshidratare medie (6-8%), iar mucoasele aparente sunt colorate normal.
Periodic (la interval de 3-5 minute) animalul prezint o stare de agitaie,
manifestat prin ridicarea capului, nsoit de polipnee, care nceteaz dup 1-2
minute. n regiunea perineal este prezent o tumefacie paraanal dreapt, de
mrimea unei jumti de portocal, anusul este deformat i deviat spre stnga,
iar mucoasa rectal este vizibil la examenul clinic din apropiere. La examenul
transrectal se constat devierea lateral stng a rectului, neputndu-se preciza
alte aspecte legate de cauza tumefaciei. Pielea din regiunea tumefiat este
integr, iar consistena tumefaciei este crnoas, asemntoare muchilor i
nedureroas (fig. 1). Glandul penian este prolabat i congestionat, probabil prin
compresia i staza produs de vezic pe vasele ce irig penisul (fig. 2).
Temperatura rectal a animalului este de 38C, frecvena cardiac 180
contracii/minut, iar fecvena respiraiei este de 65/minut.
Corelaia ntre datele anamnetice i istoricul afeciunii, precum i a
informaiilor obinute la examenul clinic au condus la diagnosticul prezumtiv de
hernie perineal a vezicii urinare. Dup Brissot, H.P. prin retroflexia vezicii urinare
este o H.P. complicat, iar dup Ferreira este o urgen medical.
ntruct hernia perineal a vezicii urinare (prin retroflexia acesteia) a fost
nsoit de anurie i deci insuficien postrenal, am considerat oportun

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intervenia chirurgical de urgen pentru repunerea vezicii urinare n poziie


fiziologic (n cavitatea abdominal) i refacerea diafragmei pelvine.

Fig.1. Hernie perineal unilateral dreapt


Fig. 2. Congestia i prolapsul penisului
(aspect clinic)

Preoperator, cu 2 ore nainte de intervenie, sacii anali au fost vidai (rectul


nu coninea fecale) i am aplicat animalului o linie de perfuzie intravenoas (i.v.)
administrnd soluie de glucoz 5%, hidrocortizon hemisuccinat 25 mg i.v.,
atropin 1% - 0,4 ml subcutanat (s.c.), Etamsilat 2 ml intramuscular (i.m.),
vitamina C sol. 10 % - 8 ml (i.v.), vitamina B-complex (Vitamin B-complex-Veyx)
4 ml i.v.
S-a utilizat neuroleptanalgezia prin administrarea injectabil i.m. a xilazinei
n timpul I i a ketaminei n timpul II (la 15 minute interval, i.v.), infiltraie pe linia
de incizie cu procain 2 %.
Pe masa de operaie am poziionat animalul n decubit lateral stng, cu
membrul posterior stng (dedesubt) deplasat caudal pn se formeaz un unghi
de 45 cu membrul posterior drept. Att membrele anterioare ct i cele
posterioare se fixeaz la masa chirurgical. Coada se trage dorsal i se fixeaz,
de asemenea la mas (fig. 3). n etapa urmtoare am practicat cateterismul
vezical cu scopul de a goli vezica urinar pentru a confirma diagnosticul de
H.P.V.U. ntruct nu am reuit cateterizarea, am decis nceperea interveniei,
evitnd efectuarea cistocentezei.
Pe toat perioada interveniei chirurgicale, animalul a fost perfuzat i.v. cu
soluie de ser fiziologic (1 pictur/ sec.).
O incizie uor curb lateral este aplicat peste masa perineal tumefiat,
de la baza cozii pn la baza masei herniate, avnd grij s nu fie prea profund,
pentru a proteja coninutul acesteia. Apoi am dilacerat bont aderenele dinte piele
i sacul herniat (fig. 4). Diagnosticul prezumtiv de H.P.V.U. devine n acest
moment definitiv. Am secionat peritoneul parietal evideniind vezica urinar
retroflexat n totalitate (fig. 5). Peretele vezical avea un aspect hemoragic (cistit
hemoragic) (fig. 6).

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Fig. 4. Incizia cutanat i evidenierea


Fig. 3. Contenia animalului pe masa de
sacului herniar seros cu vezica urinar i
operaie
desprinderea aderenelor

Fig. 5. Deschiderea sacului herniar seros Fig. 6. Vezica urinar retroflexat

Dup acest timp operator am rencercat cateterizarea vezicii urinare, de


data aceasta cu succes (probabil din cauza scderii presiunii asupra vezicii, s-a
permeabilizat zona retroflexat), extrgnd aproximativ 250 ml urin uor
hemoragic (fig. 7). Dup vidare am repoziionat vezica urinar n cavitatea
abdominal, prin defectul muscular perineal (fig.8, 9). Marginile peritoneului
parietal deschis au fost unite i prinse ntr-o pens Kocher i apoi am rsucit
pensa i peritoneul pn la nchiderea orificiului hernial. Apoi am aplicat o ligatur
transfixic simetrica (8) la baza helixului creat de rsucirea peritoneului pentru a-l
nchide definitiv, cu mtase chirurgical nr. 8 (fig. 10). Excesul de peritoneu a fost
excizat. Bontul peritoneal a fost nfundat i apoi am practicat nchiderea defectului
muscular al diafragmei pelvine (fig. 11).
Astfel am unit prin aplicarea a 2-3 fire de sutur, n fire separate (Maxon 3-
0), m. sfincter anal extern cu m. obturator, apoi ntre m. coccigian i m. sfincter
anal extern i o ultim sutur ntre ligamentul sacrotuberos i m. sfincter anal
extern (fig. 12).

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Fig. 7. Vidarea vezicii prin sondaj urinar Fig. 8. Vezica urinar dup vidare

Fig. 9. Aspectul conductului pelvin herniar Fig. 10.Torsionarea sacului herniar seros
dup repunerea vezicii n poziie fiziologic i sutura transfixic a acestuia

Fig. 12. Sutura diafragmei pelvine


Fig. 11. Bontul peritoneal restant
musculare

Grsimea i pielea n exces au fost excizate. esutul conjunctiv subcutanat


a fost suturat n surjet continuu cu fir sintetic absorbabil Maxon 3-0 (fig. 13). Apoi
am mai aplicat trei puncte de sutur n fire separate prin sutura anterioar a

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esutului conjunctiv, dar mai profund pentru a prinde planul inferior al musculaturii
diafragmei pelvine. Pielea a fost suturat de rutin (cu mtase nr. 8 n fire
separate), dup care s-a administrat spray cu antibiotice (oxitetraciclin) (fig. 14).

Fig. 13. Sutura esutului conjunctiv


Fig. 14. Sutura pielii
subcutanat i adipos

Imediat, postoperator am administrat pacientului antibiotice (amoxicilin i


enrofloxacin) i vitamina C, parenteral. Spasmium (caroverina i novaminsulfon)
0,5 ml i.m. ca spasmolitic, analgezic i antiinflamator, i 20 ml soluie 5 % KCl per
rect preparat extemporaneu, pentru a corecta hipokalemia care se instaureaz
imediat relurii tranzitului urinar consecutiv unei obstrucii urinare.
Pacientul a fost monitorizat timp de cinci zile postoperator, examinndu-se
starea general, aspectul plgii chirurgicale i eventualele complicaii renale. n
acest interval cinelui i s-au administrat antibiotice (amoxicilin i enrofloxacin),
vitamine (complexul AD3E, C, B-complex), lichide hidroenergoelectrolitice
(glucoz 5%, ser fiziologic), pansamente gastro-intestinale (Smecta),
spasmolitice, analgezice i antiinflamator (Spasmium), Hidroxizin (sedativ i
antipruriginos).

Fig. 15. Hrnirea cu alimente lichide, Fig. 16. Aspect clinic al plgii operatorii la
administrate prin sond nazo-gastric 30 zile postoperator

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La 24 ore dup operaie, animalului i-a fost introdus o sond alimentar


naso-gastric pentru hrnirea animalului cu alimente lichide, n vederea prevenirii
formrii de fecale voluminoase i a eforturilor de defecare (fig. 15). Sonda a fost
meninut timp de cinci zile. La sfritul perioadei de monitorizare, cinele
prezenta o stare general bun, cu apetit prezent pentru hran i ap, evoluia
plgii operatorii fiind normal, spre vindecare. Firele de sutur au fost scoase la
14 zile de la intervenia chirurgical, iar la 30 zile plaga operatorie i regiunea
perineal evoluau favorabil (fig. 16).

REZULTATE I DISCUII

Dup Bojrab, animalul se plaseaz n decubit sternal, cu trenul posterior


ridicat, iar Bruhl recomand decubitul lateral n special la animalele n vrst,
pentru c asigur o bun poziie a capului i o bun ventilaiei pulmonar prin
evitarea compresiei exercitate de viscere pe diafragm, un output cardiac i o
presiunea arterial satisfctoare, evitnd astfel posibilele complicaii anestezice.
Din aceste considerente i noi am adoptat poziionarea n decubit lateral a
animalului (fig.3).
Bojrab, Howard, Bruhl i Burtan recomand aplicarea unui tampon n rect i
a unei suturi n burs a anusului, pentru a evita eliminarea de fecale sau a
scurgerilor lichidului de clism, n timpul operaiei. Noi am considerat inadecvat
aceast manoper i chiar clisma, dac animalului i-au fost ndeprtate fecalele
din rect, evitndu-se asfel lezionarea suplimentar a acestuia.
Bruhl, Howard, Bojrab i Burtan recomand cistocenteza n caz de nereuit
a sondajului vezicii urinare, preoperator. Noi am evitat cistocenteza pentru c:
incizia pielii, debridarea peritoneului i incizia acestuia au fost mai facile de
realizat, prezentarea acestora operatorului fiind mult mbuntit; cistocenteza
poate fi un factor de contaminare a plgii operatorii; coagulii creai de hemoragia
intravezical consecutiv punciei ar putea cauza obstrucia vezicii, fapt constatat
anterior de noi n practic.
Pentru prevenirea recurenelor Bojrab i Bongartz recomand imediat dup
refacerea diafragmei pelvine, efectuarea castrrii animalului, iar Howard
recomand castrarea nainte de intervenia de refacere a diafragmei pelvine.
ntruct cinele prezenta criptorhidie bilateral, nu am putut realiza orhiectomia
acestuia prin tehnica clasic. Aplicarea tehnicii chirurgicale pentru castrarea
masculilor criptorhizi, avnd n vedere vrsta i insuficiena postrenal a
pacientului (cu creterea plasmatic a ureei i a concentraiei creatininei,
hiperkalemie, hiperfosfatemie i neutrofilie, dup Howard), precum i alte
afeciuni asociate, am considerat a fi riscant de efectuat.
Cistopexia efectuat cu acelai scop, de a preveni recurena retroflexiei
vezicii urinare, recomandat de Brissot, Howard i Burtan care implic efectuarea
laparatomiei, nu am efectuat-o din aceleai considerente.

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Refacerea diafragmei pelvine cunoate mai multe procedee chirurgicale:


tehnica clasic, prezentat n orice manual de chirurgie a animalelor mici,
presupune nchiderea diafragmei pelvine prin aplicarea unor suturi ntre m.
coccigian i m. sfincter anal extern, apoi ntre ligamentul sacrotuberal i sfincterul
anal extern i o ultim sutur ntre m. obturator ventral i m. sfincter anal extern
(3).
n tehnica prezentat de Earley i Kolata, marginea caudal a m. obturator
intern este incizat i muchiul este folosit pentru acoperirea defectului diafragmei
pelvine. O prim sutur este plasat ntre m. sfincterul anal extern i ligamentul
sacrotuberos (Howard propune suturarea m. sfincter anal extern de m. coccigian),
pentru a se crea suportul de prindere al apexului m. obturator intern. Astfel,
marginea caudo-lateral a m. obturator intern se sutureaz la marginea caudo-
medial a lig. sacrotuberos, marginea caudo-medial a m. obturator intern este
suturat de m. sfincter anal extern.
Bongartz, propune refacerea diafragmei pelvine cu ajutorul unei grefe din
fascia lata, izolat de pe coapsa ipsilateral i direcionat ctre defectul perineal.
Chambers, propune folosirea jumtii proximale a m. semitendinos care se
transpoziioneaz pentru refacerea diafragmei pelvine.
Vnuk et al. a utilizat o me de polipropilen pentru nchiderea defectului
diafragmei pelvice.
Strategia chirurgical adoptat a urmrit efectuarea celei mai simple
tehnicii chirurgicale, celei mai expeditive i raionale tehnici din punct de vedere
terapeutic, avnd n vedere terenul biologic, eliminnd manoperele operatori care
ar fi crescut timpul i traumatismul operator.
Noi am folosit tehnica clasic la care am adugat un artificiu reprezentat de
nchiderea sacului herniar seros (peritoneul parietal) prin torsionarea i ligatura
acestuia, similar tehnicii din hernia ombilical (fig. 10, 11), prin aplicarea unei
ligaturi transfixice simetrice (8). Din tehnicile prezentate anterior, doar Burtan
recomand refacerea sacului herniar seros prin sutura cu catgut n fire separate,
ceilali autori nefcnd nici o precizare n acest sens.
Astfel, pentru uurina ei, tehnica clasic de refacere a diafragmei pelvine,
aceasta implicnd i peritoneul, am considerat-o a fi mai facil i mult mai fiabil
dect celelalte tehnici pe care le-am considerat inoportune prin complexitatea lor,
crescnd timpul i traumatismul operator, dar i riscul unor complicaii
postoperatorii.

CONCLUZII

1. ntr-un cabinet veterinar privat a fost diagnosticat un caz de hernie


perineal a vezicii urinare la un cine Peckinez n vrst de 11 ani.
2. Diagnosticul prezumtiv a fost precizat prin coroborarea datelor
anamnetice cu examenul clinic general i confirmat intraoperator.

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3. Modalitate terapeutic n hernia perineal a vezicii urinare este


intervenia chirurgical de repunere a vezicii urinare n poziie fiziologic i
refacerea diafragmei pelvine.
4. Tehnica chirurgical clasic de refacere a diafragmei pelvine a fost
modificat prin refacerea sacului hernial seros prin nchiderea acestuia prin
torsiune i ligatur, aplicnd o ligatur transfixic simetric.
5. Astfel, pentru uurina ei, tehnica clasic de refacere a diafragmei
pelvine care implic i peritoneul, am considerat-o a fi mai facil i fiabil dect
celelalte tehnici pe care le-am considerat inoportune prin complexitatea lor,
crescnd timpul i traumatismul operator, dar i riscul unor complicaii
postoperatorii.

BIBLIOGRAFIE

1. Barnes R., 2004 Perineal hernias in the dog and cat., www.acvs.org/
AnimalOwners/ HealthConditiond/ SmallAnimalTopics/ Peritoneal Herniasinthe
DogandCat.
2. Bellenger C.R., 1980 Perineal hernia in dogs. Aust vet J. Sept; 56(9): 434-8.
3. Bojrab M.J., Toomey A.A., 1983 Perineal Hernia n Current Techniques in Small
Animall Surgery - Joseph Bojrab, Stephen W. Crane, Steven P. Arnoczky, Lea &
Febiger. Philadelphia pp.401-405.
4. Bongartz A, Carofiglio F, Balligand M, Heimann M, Hamaide A., 2005 Use of
autogenous fascia lata graft for perineal herniorrhaphy in dogs. Vet Surg. Jul-
Aug; 34(4): 405-13.
5. Brissot H.N., Dupre G.P., Bouvz B.M., 2004 Use of laparotomy in a staged
approach for resolution of bilateral or complicated perineal hernia in 41 dogs. Vet
Surg. Jul-Aug; 33(4): 412-21.
6. Brhl-Day R., Mangieri J., 2002 Perineal Hernia, Lateral vs. Caudal Approach,
Scientific Prezentation, 27th World Small Animal Veterinary Association
Congress, Granada, Spain.
7. Burtan I., 2000 Chirurgie regional veterinar, Ed. Ion Ionescu de la Brad, Iai.
8. Cazacu P., Mare M., 2004 Hemostaza cu un nou tip de ligatur transfixic n
ovariectomie la cea i pisic, Lucr. t. Med. Vet., USAMV, Iai; 47(6): 519-526.
9. Chambers J.N., Rawlings CA., 1991 Applications of a semitendinosus muscle flap
in two dogs. J Am Vet Med Assoc. Jul 1; 199(1): 84-6.
10. Ciubar V.C., 1954 Medicina operatoare veterinar, Ed. Agro-Silvic de Stat,
Bucureti.
11. Earley D.T., Kolota J.R., 1983 Perineal Hernia in the Dog: An Alternative Method
of Correction n Current Techniques in Small Animall Surgery - Joseph Bojrab,
Stephen W. Crane, Steven P. Arnoczky, Lea & Febiger. Philadelphia pp.405-
407.
12. Ferreira F., Delgado Esmeralda, 2003 - Hrnias perineais nos pequenos
animaisPerineal hernias in small animals, Revista Portuguesa de Cincias
Veterinrias,98 (545), p. 3-9.
13. Hosgood G, Hedlund CS, Pechman RD, Dean PW., 1995 Perineal herniorrhaphy:
perioperative data from 100 dogs. J Am Anim Hosp Assoc. Jul-Aug; 31(4): 331-
42.
14. Howard B., 2001 Perineal Hernia Repair; Proceedings; Atlantic Coast Veterinary
Conference, Atlantic City, New Jersey, USA.

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15. Merchav R., Feuermann Y., Shamay A., Ranen E., Stein U., Johnston D.E.,
Shahar R., 2005 Expression of relaxin receptor LRG7, canine relaxin, and
relaxin-like factor in the pelvic diaphragm musculature of dogs with and without
perineal hernia. Vet Surg. Sep-Oct; 34(5): 476-81.
16. Mortari Carolina Ana, Rahal Canevese Sheila, 2005 Hrnia perineal em, Cincia
Rural, Santa Maria, v35, n.5, p.1220-1228.
17. Patea E., Coofan V. et al., 1985 Anatomia comparat a animalelor domestice,
Vol. II, Ed. Didactic i pedagogic, Bucureti.
18. Vnuk D., Maticic D., Kreszinger M., Radisic B., Kos J., Lipar M., Babic T., 2006
A modified salvage technique in surgical repair of perineal hernia in dogs using
polypropylene mesh, Veterinarni Medicina, 51, 2006 (3): 111117.
19. www.petsurgery.com/perinealhernias.html

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TRATAMENTUL CHIRURGICAL AL CATARACTEI


LA CINE PRIN EXTRACIA EXTRACAPSULAR
A CRISTALINULUI OPACIFIAT
SURGICAL REMOVAL OF CATARACTS IN DOGS
BY EXTRACAPSULAR EXTRACTION

Fl. BETEG, N. MATES,


A. MUSTE, L. OANA, A. TIMEN

Cataracts in dogs belong to the eye pathology. It occurs whatever


the breed and age of the dog. Its ethiology is equivocal. The clinical sign
consists in the opacities of the lens, resulting in progressive loosing of the
vision on one or both eyes. Men were preoccupied over the years to
reestablish or at least to improve the vision of the affected dog by
conservative treatment or surgery. The results of these treatments and
efforts arent always the desired ones. The efforts were made in an attempt
to reestablish (or at least to improve) the vision of mans unselfish
companion or defender, the dog, knowing the inefficacy of the medical
conservative treatment. It is only the cataract surgery that can be useful.
Although the opinion of the national and international specialists
concerning the ethiopatogeny, the opportunity for lens extraction, the
technique, the costs, the instruments and materials needed are disputed, I
view it as a justified intention because of the aspects stated above.

MATERIAL I METODA

n vederea efecturii extraciei extracapsular a cristalinului(EEC) opacifiat


au fost luate n studiu urmtoarele cazuri:
Cazul 1. Mascul, ras comun, 11 ani cataract bilateral
Cazul 2. Mascul, ras comun, 14 ani cataract bilateral
Cazul 3. Femela, ras Cocker Spaniol, 12 ani cataract bilaterala
Cazul 4. Mascul, ras Ciobnesc German, 7 ani cataract unilateral.
Aceast variant a extraciei extracapsulare a cristalinului (EEC) am
preluat-o din oftalmologia uman i prezint avantajul unei deschideri mai reduse
a camerei anterioare, deoarece n cazul EEC la cine, dup deschiderea camerei
anterioare, apare colabarea corneei i hipotonia globului ocular (6).
Preoperator animalele au fost supuse unui examen clinic, pentru operaie
fiind selectate doar animale clinic sntoase cu stare general bun, fr
afeciuni intercurente ale globului ocular.Dup ce animalele au fost examinate i
au fost selectate pentru tratamentul operator, au fost supuse unor pregtiri
preoperatorii dup cum urmeaz. Cu 3 zile preoperator am administrat un
antibiotic pe cale general, iar topic am focut instilaii zilnice cu colire pe baz de
antibiotice i antiinflamatorii. Am folosit produsul Spersadex un produs al firmei
Novartis pe baz de Cloramfenicol i Dexametazona. n ziua interveniei

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operatorii am efectuat instilatii oculare cu atropin 1% cu dou ore naintea


nceperii operaiei din 15 n 15 minute pentru producerea midriazei necesare
actului operator (3). De asemenea am utilizat si Tropicamid (Mydrium)
Anestezia. Anestezia general pentru efectuarea extraciei cristalinului a
fost de tip N-NLA (narconeuroleptanalgezie) (1, 8). Aceast anestezie am
efectuat-o astfel: prima dat am procedat la administrarea Xilazinei i.m. n doz
de 2 mg/kc, iar la 10 minute dup prima injecie am administrat Ketamina n doz
de 2-5 mg/kc. nainte de nceperea interveniei operatorii am procedat la
administrarea Tiopentalului i.v. n doz de aproximativ 11 mg/kc, administrndu-l
pn n momentul dispariiei reflexelor i apariia akineziei globului ocular (5, 8).
Pentru o akinezie a globului ocular ct mai bun i pentru o propulsare a globului
ocular din orbit am efectuat i o anestezie retrobulbar cu xilin 1%.
Contenia animalului. Pe masa de operaie s-a efectuat n decubit lateral,
capul animalului fiind fixat de ctre un ajutor astfel ncat globul ocular s fie expus
pe ct posibil orizontal pentru a se putea asigura drenajul lichidului de perfuzare a
camerei anterioare.
Timpii operatori.
Meninerea larg deschise a fantei palpebrale am realizat-o prin aplicarea
blefarostatului.
Efectuarea port-side-urilor corneene Port-side-urile corneene reprezint
dou mici bree (incizii) corneene realizate cu ajutorul cuitului de cornee (V-
lance), similare cu breele realizate la nivelul abdomenului n cazul chirurgiei
laparoscopice. Un port-side se efectueaz la nivelul orei 5 sau 6 pentru aplicarea
mainteiner-ului, iar o alta bresa se execut la nivelul orei 9 sau 10 pentru
introducerea acului discizor pentru capsulotomie.
Aplicarea mainteiner-ului. Mainteiner-ul reprezint o canul pe care l-am
introdus la nivelul port-side-ului de la nivelul 5, racordat manual printr-un perfuzor
la o sursa de ser fiziologic. Rolul su este de a perfuza continuu camera
anterioar i practic globul ocular, de a ajuta la expulzarea nucleului opacifiat,
prevenind colabarea corneei dup deschiderea camerei anterioare.
Conjunctivotomia perilimbica. Incizia conjunctivei s-a efectuat la nceput cu
cuitul oftalmologic, iar apoi s-a continuat cu foarfeca de conjunctiv, uor oblic
n raport cu limbul sclerocorneean, i la o distan de 3 mm de limb. Lungimea
inciziei conjunctivei bulbare a fost de aproximativ 120-150 de grade. Ea se
efectueaz cu scopul descoperirii sclerei n vederea deschiderii camerei
anterioare pe de o parte, ct i pentru acoperirea rnii sclerale pentru o mai bun
cicatrizare. Locul de elecie la nivelul globului ocular a fost la nivelul limbului
superior centrat pe ora 12.
Deschiderea camerei anterioare. Deschiderea am realizat-o prin metoda
sclerocorneeana. Pentru aceasta am efectuat incizia sclerei cu ajutorul cuitului
de sclera pe o lungime de 120 grade centrat pe ora 12, iar apoi cu ajutorul
cuitului Crescens (un cuit oblic) s-a continuat tunelizarea sclerei respectiv a
corneei, meninnd tot timpul cuitul cu lama i vrful orientat de jos n sus. n
momentul deschiderii camerei anterioare manoperele se vor efectua cu mare
finee pentru evitarea atingerii irisului respectiv al endoteliului corneean. Rana
sclerocorneean rezultat trebuie s fie ca i un tunel, forma ei trebuie s fie ca
i un trunchi de con pe seciune cu baza mare spre camera anterioar. Aceast

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form este foarte important pentru expulzarea cristalinului opacifiat care se va


mula etan pe marginile rnii i astfel prin presiunea creat de mainteiner prin
perfuzare s-l foreze s progreseze puin cte puin spre exterior, fenomene
similare cu cele derulate n timpul naterii, folosindu-se chiar i termenul de
naterea nucleului. Aceast metod de abord tunelizat sclerocorneean prezint
marele avantaj al unei suprafee de contact mai mare ntre cele dou buze ale
rnii i datorit acesteia pierdera de lichide camerulare sunt minime, iar
etaneitatea globului ocular n vederea cicatrizrii este bun.
Capsulotomia anterioar. Reprezint deschiderea cristaloidei anterioare n
vederea extraciei cristalinului opacifiat din sacul su. Capsulotomia a fost
realizat prin tehnica beer can opener. Capsulotomia anterioar s-a efectuat cu
ajutorul acului discizor, care este un ac de sering special pregtit i modelat.
Acul discizor se introduce n camera anterioar pe la nivelul port-side-ului de la
ora 9 sau 10. Apoi acul perforeaz capsula i efectueaz incizii radiare mici de la
periferie spre centru cu mare grij pentru a nu leziona endoteliul corneean i a nu
deira zonula Zinn care practic ar putea compromite succesul interveniei.
Hidrodisecia capsulei posterioare. Reprezint detaarea capsulei
posterioare a cristalinului n vederea luxrii cristalinului n camera anterioar.
Luxarea cristalinului opacifiat n camera anterioar. Dup ce s-a efectuat
hidrodisecia, cristalinul trebuie adus n camera anterioar n vederea extraciei
finale. Pentru aceasta am procedat la creterea debitului lichidului perfuzat, iar
nucleul cristalinului a fost luxat n camera anterioar cu ajutorul acului Annel.
Luxarea se face cu mare grij pentru a nu deira capsula posterioara.
Evacuarea nucleului cristalinian opacifiat. Dup ce cristalinul este n
camera anterioar, cu ajutorul unei pense sau a unui crlig am procedat la rotirea
lui pentru a ne asigura c nu mai exist legturi ntre nucleu i capsula
posterioar. Dup ce cristalinul a fost rotit se crete debitul de lichid n camera
anterioar astfel nct cristalinul s se angajeze la nivelul inciziei corneosclerale
spre evacuare. Dup ce cristalinul s-a angajat la nivelul inciziei cu ajutorul glide (o
bucata de plexiglas de 3cm/5mm) se depreseaz buza inferioar a inciziei pentru
a permite angajarea progresiv, concomitent cu presiuni sclerale.
Excizia resturilor de cortex i a celor capsulare Acest timp se face prin
irigarea n continuare a camerei anterioare care va duce la mobilizarea acestora
urmat de aspirarea lor cu ajutorul unei seringi prevazut cu canul. Resturile
capsulare se excizeaz cu foarteca oftalmologic curb puin cte puin.
Sutura rnii operatorii. Se refer la nchiderea rnii de la nivelul
conjunctivei bulbare n cteva puncte separate. Sutura conjunctivei s-a efectuat
cu Prolene 8.0 in 3 puncte de sutur separate cu nodurile plasate la interior
pentru a preveni iritaiile conjunctivale.
Refacerea camerei anterioare. Se face scoaterea mainteinr-ului iar apoi cu
o seringa se introduce ser fiziologic, iar la final se introduce puin aer cu rolul de a
poziiona irisul. Pe msur ce umoarea apoas se reface are loc resorbia aerului
fr alte complicaii secundare din cauza prezenei aerului. Introducerea aerului
n camera anterioar are efect benefic meninnd deschis camera anterioar, el
resorbndu-se mai greu, prezena fiind util pn la refacerea umorii apoase care
n aceast perioad postoperatorie se reface mai greu. (4, 7, 10). Zilnic animalele

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au fost examinate clinic, fcndu-se instilaii cu colir Spersadex comp. timp de 7-


14 zile postoperator pn la reducerea fenomenelor inflamatorii.

REZULTATE I DISCUII

n urma examinrii celor 4 cazuri menionate mai sus, EEC a fost efectuat
pe 3 globi oculari.
Cazul 1. Cinele prezenta o cataract bilateral, cu pierderea total a
acuitii vizuale. La examenul atent al globului ocular stng am constatat luxaia
cristalinului opacifiat in vitros, mai exact era prezent o zonuloliz total dar fr
pierdere de vitros. n urma anamnezei proprietarul ne-a spus c, cinele a suferit
un accident i n urma acestui accident au aprut opacifierile cristaliniene. Decizia
terapeutic a fost lensectomia cristalinului drept, proprietarul acceptnd
efectuarea interveniei operatorii. Diagnosticul stabilit a fost de cataract
bilateral traumatic matur, luxaie cristalinian stng.
Cazul 2. Cinele prezenta cataract bilateral, cu pierderea total a acuitii
vizuale, cu dificulti de deplasare n mediu. n urma prezentrii situaiei
proprietarul a acceptat efectuarea interveniei operatorii. n momentul examinrii
diagnosticul stabilit a fost de cataract senil bilateral.
Cazul 3. Cinele prezenta cataract bilateral, dar n urma prezentrii
anselor de reuit, a riscurilor operatorii proprietarul nu a fost deacord cu
efectuarea interveniei operatorii prefernd administrarea de colire topice.
Cazul 4. Cinele prezenta cataract unilateral dreapt matur, proprietarul
neputnd furniza date despre vreun posibil traumatism. Fiind unilateral cinele
descurcndu-se fr dificulti n mediu, proprietarul a decis amnarea
tratamentului chirurgical. Practic EEC a fost efectuat n cazurile 1 si 2, pe 3 globi
oculari, cazul numarul 2 fiind operat bilateral, cazul 1 fiind operat doar la ochiul
drept.
n ceea ce privete anestezia general a animalelor combinaia folosit
Xilazin + Ketamin, completat cu Tiopental Sodic i.v. ofer condiiile necesare
interveniei operatorii, chiar dac n literatura de specialitate se citeaz necesitatea
intubaiei i narcoza gazoas, totui metoda aplicat ne-a permis efectuarea
interveniei fr dificultai. La om EEC se efectueaz doar sub anestezie
retrobulbar i aceasta este suficient pentru confortul operator. n cadrul EEC
metoda mini-nuc, o atenie deosebit trebuie acordat conteniei, mai precis
modului de fixare al capului. Capul trebuie fixat aproximativ orizontal, cu unghiul
extern uor decliv pentru drenarea lichidului de perfuzare a camerei anterioare.
Midriaza operatorie trebuie s fie foarte bun, iar aceasta la cine spre
deosebire de om se obine destul de greu, fiind necesar administrarea
preoperatorie topic a Atropinei 1% cu cteva ore nainte de nceperea actului
operator pentru obinerea unei midriaze adecvate i durabile. n cazul 1 am
ntmpinat dificulti din acest cauz deoarece dup deschiderea camerei

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anterioare n momentul capsulotomiei s-a produs o mioza deosebit de puternic


care nu a permis continuarea interveniei. Instilaiile n acest caz au fost facute
doar cu o jumtate de or preoperator. n cazul al doilea la ochiul drept am utilizat
produsul Mydrium (tropicamida) care a fost instilat tot aa cu o jumtate de or
nainte repetndu-se aceeai mioza n momentul capsulotomiei, cu toate c
tropicamida este un midriatic foarte bun n producerea midriazei. Rezultate foarte
bune am obinut prin efectuarea de instilaii oculare cu atropin 1% cu 5 ore
preoperator, combinate cu tropicamid alternativ rezultnd astfel o midriaza foarte
bun i de durata n cazul EEC de la ochiul stng (2, 9) Postoperator, cinii au
fost inui sub observaie strict n mediu fr praf i gaze iritante timp de 14 zile.
Zilnic s-au fcut instilaii cu colir Spersadex i s-au urmrit ndeaproape evoluia
fenomenelor inflamatorii intraoculare i a reaciei locale postoperatorii.
n prima zi postoperator toate cazurile au prezentat o pronunat reacie
inflamatorie a corneei care era tulbure uor opacifiat cu nuan albstruie.
Aceast intumescent a corneei s-a observat chiar din momentul efecturii port-
side-urilor sclerale, la locul respectiv aprnd instantaneu o opacifiere localizat.
Lichidul din camera anterioar este opalescent usor tulbure. Aceste fenomene
inflamatorii i pstreaz intensitatea pn n ziua a cincea postoperator cnd ele
incep sa regreseze dar transparena corneei si clarificarea lichidului camerular
apare dupa 7-10 zile devenind aproape clare dup 14 zile postoperator. Cu aceast
ocazie acuitatea vizuala a cainelui a inceput sa se amelioreze semnificativ. La 7
zile postoperator din cauza cicatrizrii ranii conjunctivale apare pruritul iar
animalul incepe s se grateze destul de violent, iar dac nu e supravegheat ii
poate produce leziuni oculare grave. Pentru evitarea unor asemenea complicaii
nedorite proprietarii trebuiesc pregatiti in aceasta privinta, si s ne asigurm de
cooperarea lor deplin.

CONCLUZII

Operaia de extracie extracapsular a cristalinului opacifiat prin metoda


mini-nuc este o intervenie operatorie complex care necesit instrumentar,
aparatur adecvat si deprinderi chirurgicale de specialitate.
Nu toi cinii cu cataract se preteaz pentru operaia de extracie a
cristalinului opacifiat. Se justific operatia EEC doar in caz de cataract bilaterala
matur, cnd animalul nu se descurc in mediul i ii modific comportamentul.
Contenia cinilor, in mod special imobilizarea capului intraoperator trebuie
efectuat astfel inct globul ocular s fie expus orizontal n vederea unei abordri
ct mai facile.
Deschiderea camerei anterioare s se fac prin incizie sclerocorneean
tunelizat pentru c altfel intraoperator apare pierderea umorii apoase i
hipotomia globului ocular.

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Corneea este deosebit de sensibil, in momentul inciziei apare instantaneu o


intumescent localizat, iar in momentul deschiderii camerei anterioare dac nu se
asigur perfuzarea continu apare colabarea ei.
Fenomenele inflamatorii postoperatorii sunt semnificative traducndu-se
clinic prin opacifierea corneei si turbiditatea lichidului din camera anterioara care
persist chiar cu tratament antiinfectios si antiinflamator timp de 7-10 zile.
Extractia extracapsular a cristalinului opacifiat, metoda mini-nuc este
avantajoas pentru c se realizeaza o incizie sclerocorneean mai mic dect n
tehnica clasic, reducnd astfel riscul hipotoniei intraoperatorii a globului ocular.
EEC prin metoda mini-nuc este avantajoas comparativ cu extracia
intracapsulara pentru c nu duce la pierderea de vitros, iar comparativ cu
facoemulsificarea este de preferat pentru c nu necesit o aparatur att de
sofisticat, permitnd in acelasi timp efectuarea implantului de cristalin artificial
de camera posterioar.

BIBLIOGRAFIE

1. Birchard S.J., Sherding R.G., 1994 - Manual of Small Animal Practice, W.B.
Saunders Company, p.1208-1212.
2. Douglas H. Slatter, 1981 - Fundamentals of Veterinary Ophthalmology, W.B.
Saunders Company Philadelphia , London, Toronto, p.489 526.
3. Dziezyc J., 1990 - Cataract surgery-current approaches, Vet Clin North Am [Sm
Anim Pract-Sm Anim Ophthalmol] 20:737-754.
4. Gelatt K.N., 1999 - Veterinary Ophtalology, Lippincott Williams &Wilkins, p.797-857.
5. Gelatt K.N., Janice P. Gelatt, 2001 - Small animal Ophtalmic Surgery, p.286-
334.Butterworth Heinemann
6. Gwin R.M., Gelatt K.N., 1981 - The canine lens. In Gelatt KN, Veterinary
Ophthalmology. Philadelphia: Lea & Febiger, p.435-473.
7. Magrane, W.G., 1977 - Canine Ophthalmology, 3rd Ed.Lea&Febiger, Philadelphia,
p.65-84.
8. Moldovan M., Bolte S., 1984 - Oftalmologia veterinara, Ed. Ceres, Bucuresti.
9. Ronald C. Riis, 2002 - Small Animal Ophthalmology Secrets, A Hanley&Belfus
Publication, p.184-192.
nd
10. Severin, G.A., 1976 - Veterinary Ophthalmology, Notes, 2 Ed. Fort Collins,
Colorado. p.77-82, p.134-148.

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OSTEOSINTEZA COLULUI FEMURAL LA CAINE CU TIJE


DE ACID POLIGLICOLIC
FEMORAL NECK OSTEOSYNTHESIS
OF THE DOG WITH POLYGLICOLIC ACID ROD

C. IGNA, Larisa SCHUSZLER,


M. SABU

The purpose of this study was to examine the effects of polyglycolic


acid rod (SR-PGA) and their applicability in the fixation of a femoral neck
fracture in dogs. In 5 dogs a Biofix rod were insert into proximal femoral
and drive into the femoral head to fix a femoral neck fractures. The follow-
up intervals were, 4, 6, 8, 10, and 24 weeks. Solid union of the fracture site
without secondary displacement was seen radiographically 6-8 weeks after
the operation in all dogs. Persistent avascular necrosis of the femoral head
and progressive osteoarthritis does not appear as a complication in dogs
treated.

Fracturile colului femural ridic probleme deosebite, inventariate minuios


de literature de specialitate: reducerea i fixarea dificil a focarului de fractur,
colapsul gtului femurului, lezarea vaselor sangvine responsabile de nutriia
capului femural i consecutiv apariia necrozei avasculare, vicii de calusare,
osteoartrit progresiv (1-8, 10, 12-14, 17-19, 21, 22).
Avnd n vedere faptul c de o parte din dificulti sunt responsabile
mijloacele metalice (broe, uruburi, tije, plci) de fixare a focarului de fractur,
inserate n montaje de compresiune (5-7, 12, 14, 17, 22) i bazai pe studii care au
testat materialele resorbabile sintetice pentru fixarea fracturilor diafizare ale
femurului (15) am considerat util verificarea clinic a tijelor de acid poliglicolic
(Biofix Braun GmbH) pentru stabilizarea fracturilor colului femural la cine.

MATERIAL I METOD

Materialul biologic a fos reprezentat de 5 cazuri clinice, cini de talie mare


cu fracturi unilaterale ale colului femural tabelul 1, fig. 1.
Abordarea operatorie a articulaiei s-a efectuat prin acces lateral cu
osteotomia trocanterului mare al femurului (20). Intervenia chirurgical, la toi
cinii, s-a desfurat sub anestezie general de tip N-NLA (xylazin -1 mg x
kgcorp-1, ketamine - 2 mg x kgcorp-1, inducie cu thiopenthal sodic - autodozaj i
narcoz inhalatorie n circuit nchis cu halothan - 4 2%).
Dup examinarea situaiei patologice locale (amploarea distruciei capsulei,
integritatea capului femural, congruena cavitaii acetabulare) s-a efectuat
reducerea manual-instrumental a focarului de fractur.

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Tabelul 1
Materialul biologic

Diagnosticul
Nr Greutate Vrsta
Rasa Fractura colului Luxaia capului
crt (kg) (ani)
femural femural
1 Brac german 24 2 x x
2 Ciobnesc german 28 2 x -
3 Collie 22 4 x -
4 Ciobnesc mioritic 45 8 x -
5 Rottweiler 36 3 x x

Fig. 1. Radiografie
preoperatorie a cazului cu Fig. 2. Imagine intraoperatorie Fig. 3. Radiografie
nr. crt 5 fractura colului a fixrii fracturii colului femural postoperatorie a cazului
femoral cu dislocaie coxo- cu tije cu nr. crt 5.
femural

Pentru fixarea focarului s-a recurs la forarea cu un burghiu de 2,5 mm a trei


traiecte osoase transfixice dinspre capul femural spre cel de al treilea trocanter al
femurului tehnica triangulaiei (2-4, 8, 17-19, 21, 22) pentru cazurile nsoite i
de luxaia capului femural. La cele trei cazuri care prezentu numai fractura simpl
a colului femural, traiectele osoase au fost forate dinspre cel de al treilea
trocanter al femurului nspre capul femural, nepenetrnd suprafaa articular a
acestuia. n traictele forate s-au introdus forat trei broe de Biofix cu diametrul de
2,6 mm. Asigurarea compresiunii n focar s-a facut prin sertizarea termic a
capetelor broelor, conform indicaiilor productorului Braun GmbH fig. 2. n
cele dou cazuri (cu fractura colului i luxaia capului femural) s-a recurs la
capsulorafie parial urmat de protezarea cu fir de Nylon 4-6 m a marginii
craniale a capsulei articulare (9).

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Fixarea trocanterului mare osteotomizat s-a efectuat cu dou broe


Krschner i o ligatur metalic de tensionare fig. 3.
Postoperator s-a efectuat o radiografie de control, verificnd corectitudinea
reducerii i fixrii. Pentru prevenirea complicaiilor septice, timp de 5 zile, s-a
administrat cefalexin 50 mg x kgcorp-1. Un pansament de tip Ehmer a asigurat
imobilizarea suplimentar a membrului afectat, timp de 14 zile. Evoluia
postoperatorie a fost monitorizat n ambulator de ctre proprietar i de medicul
curant, fiecare individ beneficiind i de 2-4 reevaluri prin examen clinic i
radiografic n intervalul 14 180 de zile postoperator. S-a urmrit: stabilitatea
montajului, modul i calitatea vindecrii osoase, locomoia i mobilitatea
articular.

REZULTATE I DISCUII

Evoluia postoperatorie a celor cinci cazuri a fost excelent. Postoperator,


dup primele 14 zile, trei animale au prezentat o chioptur de gradul 3 i 4 iar
doi cini o schioptur de gradul 2, timp de 10-14 zile. Dup alte 10 14 zile,
locomoia a devenit normal la toi cei cinci indivizi. Nu s-a semnalat nici o
complicaie postoperatorie.
Mobilitatea pasiv a articulaiei coxofemurale operate este redobndit n
totalitate, la toi cei cinci indivizi, dup 3-4 sptmni de la operaie. La 45 de zile
postoperator, analiza mersului nu a relevat, la cazuistica luat n studiu, deosebiri
de mobilitate articular (extensie, flexie, abducie, rotaie) ntre piciorul operat i
cel integru.
Imaginile radiografice au surprins o vindecare osoas complet la 6-8
sptmni postoperator. Integritatea i corectitudinea amplasrii tijelor de acid
poliglicolic nu a putut fi surprins radiografic la nici unul din cele cinci cazuri. La
controalele radiografice efectuate la 4, 6, 8, 10 si 24 de sptmni nu au fost
surprinse modificri care s indice instalarea necrozei avasculare a capului
femural, vicii de calusare i / sau semne de osteoartrit.
Abordarea focarului de fractur prin acces lateral cu osteotomia
trocanterului mare al femurului (20) permite o expunere larg, o identificare i o
manipulare facil a fragmentelor osoase din focarul de fractur-dislocaie,
permind, fa de alte procedee (2-4, 8, 10, 18, 19, 21, 22), efectuarea reducerii i
fixrii.sub controlul vizual deplin a tuturor elementelor anatomice implicate.
Tijele de acid poliglicolic (Biofix) inserate dup tehnica triangulaiei,
practicat pentru inseria broelor metalice (4, 8, 18, 21), asigur o stabilitate
excelent a fracturilor colului femural la cine. Vindecarea focarelor de fractur
are loc cu 1-4 sptmni mai rapid dect n fixrile cu mijloace metalice (1, 3, 6,
14, 17, 18, 21, 22), confirmnd rezultatele raportate n osteosinteza cu tije
resorbabile a fracturilor diafizei femurului (15).
Absena modificrilor radiografice patognomonice pentru necroza
avascular a capului femural i osteoartrit (oz) coxofemural, nregistrat la

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cele cinci cazuri, a fost semnalat i de ali autori care au recurs ns la fixri
metalice n montaje de compresiune (3, 6, 8, 11, 16, 17), atestnd stabilitatea i
eficacitatea utilizrii broelor de acid poliglicolic pentru fixarea fracturilor colului
femural la cine.

CONCLUZII

Fixarea fracturilor colului femural cu tije de acid poliglicolic, inserate dup


tehnica triangulaiei, permite realizarea unui montaj stabil, comparabil cu cel
obinut cu mijloacele metalice, asigurnd o vindecare fr complicaii.
n fracturile colului femural la cine montajele cu tije de acid poliglicolic
asigur o vindecare mai rapid a dect cea obinut prin fixrile cu materiale
metalice.

BIBLIOGRAFIE

1. Beale B., 2004 - Orthopedic clinical techniques femur fracture repair. Clin Tech Small
Anim Pract. 19 (3): 134-50. Pub. Med. Index: 15712460
2. Bojrab M.J., 1990 - Current Techniques in Small Animal Surgery. 3rd ed.,
Philadelphia, Lea & Febiger, 682-685
3. Brinker W.O., Piermattei D.L., Flo G.L., 1990 - Handbook of Small Animal
Orthopaedics and Fracture Treatment. 3rd ed., Philadelphia, W. B. Saunders,
486-488
4. Brinker W.O., Olmstead M.L., Sumner-Smith G., Prieur W.D., 1998 - Manual of
Internal Fixation in Small Animals. 2nd ed., Springer, 155-158
5. Brown B.C., 1972 - Femoral neck fracture in a dog. J Am Vet Med Assoc., 1;161 (7):
800-2. Pub. Med. Index: 5074262
6. DeCamp C.E., Probst C.W., Thomas M.W., 1989 - Internal fixation of femoral
capital physeal injuries in dogs: 40 cases (1979-1987). J Am Vet Med Assoc. Jun
15; 194 (12): 1750-4, Pub. Med. Index: 2753801.
7. Engel E.E., Volpon J.B., Shimano A.C., 1997 - Mechanical testing of the tension
band wire fixation in the proximal femur. Arch Orthop Trauma Surg., 116 (5):
266-70. Pub. Med. Index: 9177801
8. Fossum Theresa Welch, 1999 Manual of Small Animal Surgery. Ed. Mosby, St.
Louis, 689-691.
9. Igna C., 2005 - Contribuii la tratamentul chirurgical al luxaiei coxo-femurale. Lucr.
t. USAMVB Timioara, Med. Vet., vol XXXVIII, 238-241.
10. LEplattenier H.F., Montavon P.M., 1997 - Avulsion fractures of the femoral head:
internal fixation using a ventral approach to the hip joint. VCOT, 1: 23-26
11. Kaderly R.E., Anderson W.D., Anderson B.G., 1982 - Extraosseous vascular
supply to the mature dogs coxofemoral joint. Am J Vet Res, 43: 1208-1214.
12. Kuzma A., Sumner-Smith G., Miller C., Mc Laughlin R., 1989 - A technique for
repair of femoral capital epiphyseal fractures in the dog. J Small Anim Pract, 30:
444-448
13. Matis U., Waibl H., 1985 - Proximal femoral fractures in cats and dogs. Tierarztl Prax
Suppl, 1: 159-78. Pub. Med. Index: 4012774
14. Miller A., Anderson T.J., 1993 - Complication of articular lag screw fixation of
femoral capital epiphyseal separations. J Small Anim Pract 34: 9-12

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15. Miettinen H., Makela E.A., Rokkanen P., Tormala P., 1992 - Fixation of femoral
shaft osteotomy with intramedullary metallic or absorbable rod: an experimental
study on growing dogs. J Biomater Sci Polym Ed., 4 (2): 135-43. Pub. Med.
Index: 1336673
16. Moon M.S., Cho D.Y., 1980 - Trochanteric intra-osseous venography in femoral neck
fractures in the dog. Int Orthop.; 3 (4): 265-9. Pub. Med. Index: 7399765
17. Necas A., Dvorak M., 2003 - Surgical treatment of a saggital intraarticular femoral
head fracture with coxofemoral dislocation in two mature dogs. Acta Vet. Brno,
72: 261265.
18. Nunamaker D.M., 1973 - Repair of femoral head and neck fractures by
interfragmentary compression. J Am Vet Med Assoc., 1, 162 (7): 569-72. Pub.
Med. Index: 4692824
19. Olmstead M.L., 1995 - Small Animal Orthopaedics. Mosby, 228-230
20. Piermattei D.L., Greeley R.G., 2003 - An atlas of surgical approaches to the bones
th
of the dog and cat. Ed. Ed. W.B. Saunders Comp., Philadelphia, 4 ed., 300-
305.
21. Slatter D.H., 1993 - Textbook of Small Animal Surgery. Vol. 2. Philadelphia, W. B.
Saunders, 1805-1808.
22. Vernon F.F., Olmstead M.L., 1983 - Femoral Head Fractures Resulting in
Epiphyseal Fragmentation: Results of Repair in 5 Dogs. Vet Surg, 12: 123-125

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INCIDENA I CONDUITA TERAPEUTIC


N PARODONTOPATII LA CINE
INCIDENCE AND TERAPEUTICAL GUIDANCE
PARODONTOPHATY IN DOGS

A. MUSTE A., N. MATE, L. OANA,


A. TIMEN, Fl. BETEG, C. OBER
U.S.A.M.V. Cluj-Napoca

Parodontal diseases are frequently seen in dogs. From twenty dogs,


fourteen of them were diagnosed with parodontal diseases.In early stages of
parodontal diseases surgical removal of necrotic tissues was performed. In
deep tissues infections the surgical method is more complex using mucosal
membranes to cover the affected area. Complete healing depends on the
surgical technique and the quality of the mucosal membrane from
surrounding area.

La specia canin parodontopatiile sunt frecvent ntlnite, cu repercursiuni


directe asupra strii de sntate. n catalogarea i aprecierea acestor stri, trebuie
s inem cont de particularitile speciei, de tipul masticaiei, felul alimentelor,
regimul alimentar, igiena bucal, tulburri endocrine, boli metabolice, perturbri
i maladii sanguine.
Cauzele parodontopatiilor sunt cu preponderen de origine alimentar,
traumatic i accidental (prin lezionarea tisular bucal) urmat de contaminare
i suprainfectare cu germeni patogeni. Adesea suprainfectarea determin formarea
de granuloame dentare sau pungi gingivale cu urmri directe asupra integritii
esutului osos.
Cauzele apariiei, terenul biologic, incidena acestora i manifestrile
clinice, evoluia, prognosticul, reuita terapiei sunt aspecte deosebit de complexe
tocmai prin faptul c n derularea acestor entiti intervin o serie de factori
favorizani sau determinani. n aceast lucrare am ncercat s urmrim unele
aspecte privind incidena i terapia parodontopatiilor.

MATERIAL I METOD

Cazuistica luat n studiu a fost format dintr-un numr de 20 cini de rase


i vrste diferite (tabel 1) prezentai la Clinica de Patologie i Clinic Chirurgical
a Facultii de Medicin Veterinar Cluj-Napoca, avnd o simptomatologie de
disconfort bucal.
Din cei 20 de cini examinai la 14 dintre ei s-au identificat leziuni de
parodontopatii. La examenul clinic efectuat individual s-a apreciat prezena sau
absena acestor leziuni, localizarea acestora i modificrile locale existente. La
toate cazurile luate n observaie poriunea din zona afectat prezenta semne de

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inflamaie, tumefiere, edem, esuturi devitalizate, pungi parodontale i resorbie


osoas n diferite grade.
Tabelul 1
Incidena parodontopatiilor la cine

Nr.
Rasa Sex Vrsta Parodontopatii
Crt.
1. Ras comun Mascul 8 +
2. Ciobnesc german Mascul 9
3. Tekel Femel 2,5
4. Rottweiler Mascul 7
5. Mastino Napolitano Femel 4 +
6. Ras comun Mascul 7
7. Saint Bernard Femel 7 +
8. Ciobnesc german Femel 4
9. Ras comun Mascul 7 +
10. Rottweiler Femel 6 +
11. Ras comun Mascul 6 +
12. Chow-chow Mascul 4 +
13. Setter irlandez Femel 6 +
14. Cocker spaniel Mascul 6,3 +
15. Caniche Mascul 5,5 +
16. Cocker spaniel Femel 6
17. Brac german Mascul 5 +
18. Brac german Mascul 4 +
19. Cocker spaniol Femel 6 +
20. Caniche Femel 9 +

Pentru cazuistica luat n studiu am folosit dou metode chirurgicale cu


scopul de a ndeprta esuturile devitalizate sau necrozate i crearea de condiii
pentru regenerarea esutului osos.
Prima metod folosit a fost chiuretajul parodontal. Acest procedeu l-am
utilizat la 6 subieci care au prezentat: pungi inactive, leziuni localizate la un dinte,
leziuni parodontale strmpte, pungi supraosoase sau ca metod alternativ n
parodontitele severe. Cinii sunt anesteziai prin neuroleptanalgezie, dup care
sunt aezai n decubit lateral sau sternoabdominal. Chiuretajul const n
eliminarea feei interne a peretelui pungii parodontale cu ajutorul unei chiurete.
Consecutiv se ndeprteaz tartrul subgingival, cementul compromis, se elimin
esuturile inflamate i se realizeaz surfasajul radicular. Prin aceste manopere se
urmrete ndeprtarea pungii parodontale i obinerea unui ataament nou.
A doua metod folosit este tot chirurgical i este cunoscut ca operaie
cu lambou. Principiul metodei const n descoperirea osului alveolar prin
decolarea lambourilor mucoperiostale, ndeprtarea elementelor patologice,

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chiuretajul pungilor subgingivale sau osoase, nivelarea osului, urmate de sutur.


Este metoda care se folosete n cazul pungilor osoase adnci situate la un dinte,
la un numr redus, sau generalizate. Aceast metod s-a aplicat la 8 subieci din
cei 14 luai n tratament.
Tehnica operatorie const n efectuarea a dou incizii vertical oblice pe
versantul vestibular i oral de o parte i alta a grupului de dini care vor fi operai
unite printr-o incizie orizontal de-a lungul marginei libere gingivale, secionndu-
se papilele interdentare, care de obicei se gsesc tumefiate n urma procesului
inflamator. n acest fel inciziile formeaz un trapez, cu baza spre fundul de sac
vestibular. Inciziile de pe versantul vestibular merg n general pn la limita
mucoasei fixe, iar inciziile din bolta palatin vor fi scurte pentru a nu seciona
vasele. n situaiile n care se face intervenia pe toat arcada ntr-o edin se
traseaz nti incizia marginal orizontal de la un capt la cellalt al arcadei i
apoi dou incizii vertical-oblice, de o parte i de alta, n dreptul caninilor sau
primilor premolari n raport cu forma arcadei. Cu ajutorul unui decolator fin se
desprind lambourile gingivoperiostice i se rstoarn vestibular i oral.
Decolarea trebuie fcut cu manopere blnde, pentru a nu perfora lamboul.
Lambourile vestibulare sunt meninute ndeprtate cu deprttorul cu gheare,
descoperindu-se astfel osul alveolar pe ntinderea necesar pentru a se avea sub
control pungile osoase.
Cu chiurete de diferite mrimi se chiureteaz toate fongozitile i esutul
de granulaie, insistnd n mod cu totul deosebit n pungile osoase, de-a lungul
rdcinilor dentare. Se face apoi netezirea osului, aceast regularizare urmrete
transformarea atrofiei verticale ntr-o atrofie orizontal, respectiv obinerea unei
margini alveolare uniforme, netede, cu pante line n jurul dinilor desfinndu-se
pungile osoase n jurul rdcinilor. In finalul manoperelor osul trebuie s rmn
curat, neted, fr poriuni necrotice, fr proeminene i anfractuoziti, iar
pungilor de osteoliz periarticular s li se dea forma de plnie foarte evazat,
care s nu favorizeze nici un fel de retenie. Se completeaz chiuretajul de pe
poriunea decolat a lambourilor mucoperiostatice, pentru a nltura orice urm
de esut de granulaie sau de epiteliu invaginat, de pe peretele intern al fostelor
pungi paradontice. Lambourile periosteomucoase sunt readuse peste plaga
osoas i suturate cu ethicon, prolene, dexon, fire care vor fi trecute prin spaiile
interdentare.
n aceast etap trebuie avut grij s nu rmn spaii moarte care s
favorizeze formarea de colecii, motiv pentru care periosteomucoasa va trebui s
se adapteze intim la suprafaa osoas.
Chiuretajul gingival precum i operaia cu lambou se pot asocia n
tratamentul parodontopatiilor marginale n raport de forma leziunilor i topografia
acestora. Astfel la nivelul poriunii de arcad, n care pungile nu depesc 2-3
mm, se face chiuretajul chirurgical, iar n poriunile n care pungile depesc 3-4
mm se poate face fie gingivectomia fie operaia cu lambou. Cteva zile (5-7)
postoperator la nivelul inciziilor gingivale se aplic o medicaie bazat pe
antibiotice i antiinflamatoare (mibazon unguent, Hidrocortizon unguent, Asocilin
unguent, Oxi-vet spray, Oximanirom pulvis). Dup aproximativ 8-10 zile, firele se
scot, iar mucoasa trebuie s fie complet vindecat.

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REZULTATE I DISCUII

Aa cum s-a menionat, la tehnicile operatorii interveniile chirurgicale de


lambou dau rezultate foarte bune atunci cnd esutul osos afectat este ndeprtat
complet pn la esutul sntos, urmat de aplicarea unei medicaii specifice. n
cazuistica noastr din cei 12 cini la 10 dintre ei am practicat operaie radicular
prin utilizarea lamboului chirurgical, iar la 2 exemplare a fost nevoie doar de
chiuretaj chirurgical. Intervenia chirurgical a decurs normal, iar postoperator nu
am ntreprins complicaii cu excepia a 2 cazuri la care s-a produs deschiderea
rnii cu decolarea lamboului. Acest neajuns s-a remediat prin nlocuirea firelor
vechi. Considerm c este nevoie de atenie la afrontarea lambourilor decolate
pentru chiuretajul osos i realizarea unei hemostaze ngrijite n timpul actului
operator.

CONCLUZII

Leziunile inflamatorii i necrotico-supurative sunt prezente att la nivelul


esuturilor moi ct i la nivelul esuturilor dure ale cavitii bucale indiferent de
vrst (tabel. 1), regim alimentar sau stare de ntreinere, caracterul profilactic
avnd o importan mult mai nsemnat.
Parodontopatiile marginale sau vestibulare mandibulare sau palatine pot fi
tratate chirurgical prin diferite metode sau tehnici cu respectarea a cel puin dou
criterii: acela al ndeprtrii n totalitate a esutului necrozat, devitalizat i
acoperirea complet cu lambou a cmpului operator.
Pentru o vindecare complet i ntr-un timp relativ scurt recomandm ca
inciziile care se practic s fie scurte i paralele pe direcia de cretere a
miofibrilelor.
La nchiderea rnii postoperatorii, se va evita formarea de hematoame i
buzunare care s duc la acumularea de secreii.
n funcie de talie, starea de sntate a lambourilor, lipsa de substan.
distana dintre firele de sutur va fi mic (2-4mm) iar grosimea firelor potrivit.

BIBLIOGRAFIE

1. Baredstein A., Nilveus R., Egelberg J., 1985 Effect of nonsurgical periodontal
therapy. Bleeding, suppuration and probing depth in sites with robing
attachement loss. J. Clin. Periodont 12: 432 440.
2. Bercy P., Tenembaum , 1996 Paradontologie. Du diagnostic la practique.
Bruxelles: De Boeck Universit.
3. Haryey C.E., Emily P.P., 1993 Small Animal Dentistry. Philadelphia, CY Mosby
Co.
4. Holmstrom S.E., 1989 Periodontal disease, Compendium 11: 1485.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

5. Lamont McNeil R., Somerman M., 2000 Development and regeneration of the
periodontium: parallels and contrasts Periodontology, Vol. 19: 8- 20.
6. Morgan J.P., Miyabayaschi T., Anderson J., Klinge B., 1990 - Periodontal bone
loss in the aging bucale dog a radiographia study, J Clin Periodont 17: 630.
7. Muste A., A. Serbnescu, P. Magler, 2003 Implantul dentar la cine, Simpozion
tiinific Internaional 70 de ani ai Universitii Agrare de Stat din Moldova,
Chiinu 7-8 octombrie 2003, pag. 112-113.
8. Muste A., N. Mate, L.Oana, A. Timen, Fl. Beteg, 2004 Tratamentul edentaiei la
cine, Simpozion tiinific Timioara , Vol.XXXVII, pag. 546.
9. Polimeni G., Albandar J.M, and Wikesju M.E., 2002 Prognostic factors for
alveolar regeneration: osteogenic potential of rezistent bone. Journal of clinical
Periodontology 31 (10) pag. 840 844.

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TRANSPOZIIONAREA CANALULUI STENON CU


DESCHIDEREA ACESTUIA N SACUL CONJUNCTIVAL
STENON CHANNEL TRANSPOSITION IN THE CONJUNCTIVAL REGION

L. OANA, A. TIMEN, A. MUSTE,


N. MATE, Fl. BETEG

Keratoconjunctivitis sica in not frequently seen in animals.In dogs


5% of cases with eye problems have also this disease. Tear secretion may
be totally reduced causing keratitis and blindness.
The paper presents the surgical procedure consisting in Stenon
chanel transposition in the eye region. Surgical procedure was performed
in one dog with keratoconjunctivitis sica and the results were very good.

MATERIAL I METOD

Operaia chirurgical s-a efectuat pe un cine, mascul de ras comun


avnd vrsta de 5 ani, prezentnd simptome ale cheratoconjunctivitei uscate
cronice bilaterale, care a fost tratat cu colire i unguente oftalmice dar nu ntr-un
mod susinut, ci doar sporadic, ceea ce nu a dus la nlturarea consecinelor
dezastruoase ale uscrii polului anterior a globului ocular.
Pacientul prezenta urmtoarele semne clinice: corneea s-a uscat progresiv,
aparnd vlurit, acoperit de secreii uscate, micrile pleoapelor fiind dificile.
Aceasta a dus la iritarea suprafeei corneei care mai trziu a dezvoltat pe
suprafaa sa , n mod normal transparent, vase de snge i pigmeni.
Tabolul clinic, alturi de testul Schirmer au permis stabilirea diagnosticului
de cheratoconjunctivit ursac.
Pentru testul Schirmer am folosit o fie de hrtie de filtru Whattmann
N=41, lat de 5 mm i lung de 35 mm. La extremitate, am ndoit 5 mm folosind o
pens din material plastic, ne fiind indicat a se folosi degetele. Partea ndoit a
hrtiei de filtru am introdus-o sub pleoapa inferioar i am lasat-o timp de un
minut. Interpretarea se face citindu-se umezirea pe fia de hrtie de filtru.
Umezirea pe o lungime mai mic de 9 mm este considerat
nesatisfctoare, ntre 9 mm i 22 mm cu media de 14 mm este normal, iar
peste 22 mm este excesiv.
Rezultatul nostru la testul Schirmer a fost de 6 mm, fapt ce dovedete
insuficiena de secreie lacrimal.
Tehnica operatoare
Locul de elecie este reprezentat de regiunea maseterin, pe zona de
proiecie a canalului Stenon.
ncizia pielii se ntinde pe o distana de 6 cm, la mijlocul distanei dintre
glanda lacrimal i papila canalului Stenon care se deschide la nivelul molarului 3
superior.

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esutul conjunctiv subcutanat a fost dilacerat cu ajutorul unei foarfeci


chirurgicale boante, evideniindu-se fascia muchiului maseter, adiacent se
gsete treimea mijlocie a canalului Stenon.
Canalul Stenon este situat ntre nervul facial i nervul bucal dorsal.
Diferenierea se face destul de greu, deoarece au aceeai grosime, fiind totui
difereniate prin coloraie astfel, canalul Stenon are o culoare alb-roz, pe cnd
nervii au o culoare alb.
Dilacerarea canalului Stenon se face cu greutate deoarece este bine ataat
de fascia muchiului maseter. De asemenea se evit lezionarea formaiunilor
nvecinate cum ar fi nervii bucali i multiplele ramuri ale venei faciale. n
continuare se izoleaz n sens oral canalul Stenon, pn la papila ce se
deschide n cavitatea bucal, apoi am secionat mucoasa bucal n jurul papilei.
Dup ndeprtarea papilei, plaga din mucoasa bucal se sutureaz.
Trebuie acordat o mare grij poriunii terminale a canalului din cauza
traiectului su prin submucoasa cavitii bucale, traiect ce poate avea o lungime
de 0,5 1,0 cm.
Urmtoarea etap a operaiei este tunelizarea subcutanat i crearea
noului traiect, astfel cu o sond butonat se construiete un tunel subcutanat
pn n sacul conjunctival.

Sonda este mpins cu presiune n esutul subcutanat pn apare la nivelul


mucoasei conjunctivale, fr a o penetra.
Cu ajutorul bisturiului efectum o mic incizie desupra sondei butonate
crend o deschidere n care se aduce captul canalului Stenon.
Cu ajutorul unui fir de sutur se prinde papila canalului Stenon pentru a fi
dirijat mai uor spre deschiderea din sacul conjunctival. Papila i canalul sunt
dirijate prin tunelul subcutanat creat, pn la apariia papilei canalului Stenon, la
nivelul mucoasei conjunctivale. Papila se sutureaz la mucoas cu ajutorul a trei
fire de sutur (Vicryl 2.0 ).
Plaga operatorie de la nivelul zonei maseterine se sutureaz n dou
straturi. nti fascia superficial a muchiului maseter cu fire resorbabile i apoi
pielea cu a chirurgical.

REZULTATE I DISCUII

Postoperator am urmrit evoluia strii generale a animalului, vindecarea


plgilor ct i evoluia cheratoconjunctivitei uscate.

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I-am administrat pe cale general antibiotice timp de trei zile i am urmrit


evoluia procesului de vindecare a plgilor. Zilnic am efectuat toatetarea marginii
palpebrale pentru ndeprtarea secreiilor i a crustelor formate, aplicnd n sacii
conjunctivali unguent oftalmic pe baz de kanamicin i hidrocortizon.
Secreia salivar a glandei parotidiene se putea observa cu uurin la
nivelul globului ocular, mai ales n timpul mesei, crend falsa impresie de epifor,
cile lacrimale nereuind s canalizeze ntreaga cantitate de secreii.
Tratamentul la nivelul sacilor conjunctivali s-a continuat timp de o lun ,
pn la revenirea transparenei corneei i dispariia complet a semnelor de
cheratoconjunctivit.
Tratamentul medicamentos n cheratoconjunctivita uscat trebuie s
identifice n primul rnd, cauzele care au determinat boala. n prima perioad se
recomand tratament medicamentos simplu, care urmrete lubrefierea i
reumezirea corneei. Se va alege un unguent oftalmic cu pilocarpin pentru a
activa secreia lacrimal, la care se adaug vitamina A uleioas i se poate alterna
cu instalaii de soluie 20% de acetilcistein, de 4-5 ori pe zi, cte 3 picturi.
Se va institui i un tratament general cu vitaminele B1, B2, C, A , D, E, n
caz de infecie se aplic o dat la 2-3 zile unguent iodoformat 3-4%.
Terapia medical la a cheratoconjunctivitei acute include substitueni de
lacrimi , msur alternativ de tratament ce are inconvenientul frecvenei mari cu
care trebuie aplicate acestea (aproximativ 4-6 ori pe zi ). O alt abordare a
tratamentului medicamentos este stimularea secreiilor lacrimale deja existente,
pentu a crete umiditatea ochiului, folosind unguente cu cyclosporine
(Optimmune) de dou ori pe zi, timp de mai multe sptmni (Kirk i col. 1995).
Administrarea oral a pilocarpinei (de 1- 3 ori pe zi) poate stimula
producia de lacrimi, dar pilocarpina cauzeaz hipersalivaie i uneori emez i
diaree.
Cauzele acestei boli sunt mai greu identificabile de aceea tratamentul
vizeaz n special nlocuirea lacrimilor mai degrab dect corectarea cauzei.
Postoperator nu s-au observat disfuncii n masticaie, deglutiie i digestie.
Complicaiile ce ar putea s apar dup efectuarea operaiei sunt
secionarea i rsucirea n jurul propriului ax a canalului Stenon. Rsucirea
canalului Stenon cu mai mult de 180 se manifest prin secreie intermitent,
redus sau absent.
O consecin a transpoziionrii canalului Stenon poate fi apariia la nivelul
globului ocular, la nivelul conjunctivei, al pleoapelor, a unui precipitat cristalin de
sruri minerale ( carbonat de calciu, fosfai, oxalai sau o combinaie dintre
acetia). Se pare c apariia acestui precipitat este strns legat de hiperactivitatea
glandei parotide (5). La cazul nostru nu s-a identificat nici un fel de precipitat.

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CONCLUZII

n cheratoconjunctivitele acute se aplic iniial un tratament medicamentos,


care const n introducerea n sacul conjunctival a unor produse ce urmresc
lubrefierea i reumezirea corneei;
n cheratoconjunctivitele cronice, datorit inconvenientelor tratamentului
medicamentos, se indicat transpoziionarea canalului Stenon la nivelul mucoasei
conjunctivale;
Datorit similitudinii compoziiei secreiei lacrimale cu cea a salivei
parotidiene, saliva este foarte bine tolerat de ctre mucoasa conjunctival i
cornee. Dei unii autori consider c saliva i lichidul sinovial pot da reacie de
corp strin n organism, la nivelul mucoasei conjunctivale noi nu am sesizat nici
un fel de reacie, dect cea dat de traumatismul operator;
Faptul c intervenia operatoare se execut fr necesitatea unei aparaturi i
materiale speciale, preferm tratamentul chirurgical n cheratoconjunctivita uscat
fa de cel medicamentos care trebuie aplicat de 4-6 ori pe zi;
Nu am constatat modificri cauzate de aportul insufucient de saliv,
deglutiia, masticaia i toate procesele digestive i metabolice desfurndu-se
normal.

BIBLIOGARFIE

1. Betts, D.M., 1977 - The surgical correction of parotid duct transposition failures, J.
Am. Anim. Hosp. Assoc. 13: 695-700.
2. Brooks, W.C., 2002 - Dry eye, Keraconjunctivitis sicca, The Pet Health Library.
3. Gelatt, K.N., Peterson Gelatt, J., 2001 - Small Animal Ophthalmic Surgery, pg. 128-
140.
rd
4. Grahn, B.H., 1999 - Diseases and surgery of the canine nasolacrimal system, 3
edition, Baltimore , pg.569-581.
5. Kirk, R.W. and Bonagura, J.D., 1995 - Keratoconjuctivitis sicca. Kirk's Current
Veterinary Therapy XII Small Animal Practice. p. 1231-1239. W.B. Saunders Co.,
Toronto.
6. Lavignette, A.N., 1996 - Keratoconjunctivitis sicca in a dog treated by transposition
of the parotid salivary duct. J. Am. Vet. Med. Assoc. 148: 778-786.
7. Mate, N., 2004 - Patologia chirurgical a animalelor domestice, Editura Medical
Universitar Iuliu Haieganu, Cluj Napoca, pg. 302-367.
8. Moldovan, M. Bolte, S., 1984 - Oftalmologie veterinar, Editura Ceres, pg. 193-205.
9. Moore, C.P., 2000 - Dry eye sindroms: KCS and other tear deficent disease.
Transactions North American Veterinary of 14, pg. 560-562.
10. Ognean, L., Dojan, N., Roioru, C., 2000 - Fiziologia animalelor Vol. I, Editura
Presa Universitar Clujean, pg. 54-57.

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TEHNICA PROSTATECTOMIEI TOTALE LA CINE


TOTAL PROSTATECTOMY IN DOG

L. OANA, A. MUSTE, N. MATE,


A. TIMEN, Fl. BETEG

Prostatic diseases especially the tumours are frequently seen in


dogs.Current medication can not resolve always the problems being
necessary the prostatectomy.
Total prostatectomy with urethral resection was the best procedure
applied.

Animalele de companie de cele mai multe ori sunt privite ca i un membru


al familiei. Datorit acestui fapt, afectivitatea i ngrijirea primit din partea
stpnilor precum i ataamentul fa de acetia, sunt factori care duc la atingerea
unor vrste naintate, uneori chiar impresionante. Din pcate, odat cu naintarea
n vrst apar i o serie de boli grave care la tineret sau n cazul animalelor de
rent nu se manifest clinic. Aa este cazul afeciunilor meoplazice ale prostatei la
cine, afeciune care n multe cazuri nu ced la tratament medicamentos i trebuie
intervenie chirurgical.
n cazul n care carcinomul a fost descoperit nainte de apariia
metastazelor, conduita de urmat este intervenia chirurgical cu ndeprtarea
prostatei n ntregime, fiind recomandat i un tratament cu citostatice.

MATERIAL I METOD

Operaiile s-au efectuat pe 5 cini de vrste diferite, ras comun (tabel 1)


care prezentau semne clinice specifice afeciunilor prostatei (miciuni dese i cu
dificultate, la tueul rectal s-a constatat mrirea n volum a prostatei).
Tabelul 1

Pacientul 1 2 3 4 5
Rasa comun comun comun comun comun
Vrsta 9 ani 15 ani 14 ani 11 ani 12 ani

Instrumentarul i materialele utilizate au fost cele folosite curent n


interveniile chirurgicale.
n experimentul nostru am utilizat metoda prin ablaia prostatei cu rezecia
unei poriuni de uretr urmat de anastomoza termino-terminal a acesteia.
Preoperator. Animalele au fost supuse unei diete alimentare cu 12 ore
nainte de intervenia chirurgical.
nainte de operaie am determinat timpii de sngerare i de coagulare, care
trebuie s se ncadreze n limitele fiziologice.

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Anestezie general prin neuroleptanalgezie i anestezie local.


Tehnica operatoare
Trebuie menionat de la bun nceput c aceast tehnic necesit prezena
a dou ajutoare care s aib un minimum de cunotine de medicin n general i
de chirurgie n special.
n vederea operaiei animalul a fost contenionat n decubit dorsal.
S-a efectuat laparatomie paramedian, de la nivelul cicatricei ombilicale
pn n regiunea prepubian.
Pentru vizualizarea prostatei, marginile plgii sunt ndeprtate cu ajutorul
deprttoarelor simple Farabeuf, iar masa gastrointestinal este izolat ntr-un
cmp steril i mpins cranial, fr a fi scoas din cavitatea abdominal. Acest
lucru se realizeaz cu scopul evitrii periclitrii intestinelor i de asemenea pentru
a oferi condiii de lucru mai bune. De menionat c masa intestinal, vezica
urinar i cavitatea abdominal au fost irigate periodic cu soluii fiziologice pe
toat durata interveniei operatorii.
n urmtoarea etap am evideniat vezica urinar i am introdus un cateter
uretral pentru vidarea acesteia. Dup vidarea complet, pentru evidenierea
prostatei, a gtului vezical i al uretrei am dilacerat esutul adipos periprostatic i
am ncercat mobilizarea ct mai mult a prostatei i a uretrei n sens cranial.
Dup evidenierea gtului vezical i a uretrei i vidarea vezicii urinare
cateterul se retrage uor din vezic pentru a facilita rezecia uretrei. Am aplicat
dou fire de traciune pe poriunea caudal a uretrei pentru a uura realizarea
anastomozei. Se asigur hemostaza la nivelul prostatei dup care cu ajutorul
foarfecelor chirurgicale drepte are loc ndeprtarea prostatei i a unei poriuni de
uretr. Trebuie evitat rezecia unei poriuni mari din uretr pentru a conserva ct
mai mult din gtul vezicii i din poriunea caudal a uretre (Fig. 1).
Dup ndeprtarea prostatei cateterul se redirecioneaz spre vezic.
Capetele secionate ale uretrei se afronteaz i se trece la anastomozarea lor n
puncte separate cu fir resorbabil polidioxanol mrimea 4.0. Anastomozarea s-a
fcut iniial n puncte cardinale urmnd completarea celorlalte puncte de sutur
(Fig. 2). Afrontarea trebuie fcut ct mai bine i mai corect pentru a evita
rmnerea de spaii prin care s aib loc revrsarea urinei n cavitatea
abdominal.

Fig. 1 Ablaia prostatei Fig. 2 Sutura uretrei

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Se verific nc odat dac nu exist hemoragii sau corpi strini la nivelul


cavitii abdominale dup care se face lavajul peritoneal cu soluie fiziologic i
antibiotice dup care se aplic intraabdominal un unguent cu antibiotic pentru a
mpiedica apariia aderenelor i a eventualelor infecii.
Urmtoarea etap este dat de nchiderea peretelui abdominal pe dou
straturi anatomice. Primul strat reprezentat de peritoneu i esutul muscular s-a
suturat n fir continuu cu Catgut mrimea 2.0. Aceast etap trebuie efectuat cu
mare pruden pentru a evita ciupirea unei anse intestinale sau suturarea
acesteia la peretele abdominal. Al doilea strat reprezentat de esutul conjunctiv
subcutanat i piele s-a suturat n puncte separate cu mtase chirurgical lsndu-
se loc pentru dren. Acest loc de dren este necesar pentru a evita formarea unor
colecii datorate dilacerrilor ce au loc.
Pe locul inciziei s-au fcut aspersiuni cu eter iodoformat, iar cateterul s-a
fixat printr-un punct de sutur la pielea prepuului.
Tratamentul postoperator
Postoperator animalele au fost supuse unei antibioticoterapii, la primul
pacient cu penicilin (1.000.000 U.I.) i streptomicin (1 g) zilnic, iar la ceilali cu
enrofloxacin n doz de 1 ml/10 Kg mas corporal, zilnic timp de 5 zile. De
asemenea s-a fcut lichido-terapie timp de dou zile iar cateterul a fost fixat
pentru patru zile.
Animalele au fost supuse unui regim dietetic pentru a evita coprostaza i a
reduce riscul deirrii peretelui abdominal. Dup operaie animalele au fost
termometrate zilnic timp de ase zile.
Tratamentul plgii operatorii a constat n aplicarea zilnic de eter
iodoformat n scopul grbirii cicatrizrii i prevenirii infeciei.

REZULTATE I DISCUII

Evoluia postoperatorie a pacienilor a fost n general bun, n primele patru


zile urinarea s-a efectuat prin cateter dup aceea pe cale natural. Pentru a
mpiedica nchiderea cateterului, zona abdominal a fost protejat printr-un
pansament Alfort.
La doi cini datorit faptului c n primele 2 zile postoperator au refuzat
consumul de hran, le-am administrat glucoz n perfuzie.
La un pacient, la locul plgii operatorii s-a acumulat o secreie purulent
care a necesitat nlturarea a 2 fire i realizarea drenajului.
n medie, vindecarea s-a realizat n 12 zile de la operaie, fr apariia unor
complicaii majore.
La cazurile la care examenul histopatologic pune n eviden o tumor
malign, este necesar completarea tratamentului operator cu citostatice.

CONCLUZII

Interveniile chirurgicale pe prostat n practica medical veterinar se fac


rar i necesit respectarea cu strictee a unor reguli operatorii specifice.
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n urma rezultatelor obinute dup operaia de prostatectomie total am


ajuns la urmtoarele concluzii:
1. Deoarece operaia nu necesit aparatur i instrumentar special aceasta se
poate executa n bune condiii n orice clinic veterinar, cu condiia respectrii
tehnicii operatorii i a tratamentului postoperator specific operaiei de
prostatectomie.
2. n urma acestui experiment s-a constatat c uretra prezint o capacitate
mare de cicatrizare. n urma rezeciei i a anastomozrii ei, vindecarea plgii
uretrale s-a realizat suficient de bine n 12 zile de la operaie.
3. Trebuie evitat rezecarea unei poriuni prea mari din uretr pentru a
reduce riscul apariiei unor complicaii postoperatorii, amintind aici riscul cedrii
suturilor uretrale care duc la uroperitoneu i peritonit, acestea fiind unele dintre
cele mai grave. De asemenea trebuie asigurat un tratament postoperator adecvat n
special cel cu antibiotice i cateterizarea uretrei timp de cteva zile pn cnd se
realizeaz o cicatrice uretral suficient de rezistent.
4. Rezultatele obinute n urma operaiei de prostatectomie total arat c
aceast tehnic aplicat corect duce la rezultate bune i la reducerea riscurilor
majore pentru pacient. Datorit faptului c este o metod radical, duce la
reducerea riscurilor de recidiv, pretndu-se efectuarea ei i n condiii de teren
deoarece nu necesit o aparatur sofisticat, instrumentar special i condiii
deosebite. n unele situaii tratamentul poate fi completat cu medicaie citostatic.

BIBLIOGRAFIE

1. Bbasinger R.R., Rawlings C.A., 1987 - Surgical management of prostatic disease,


Comp Cont Educ Pract Vet 9:993.
2. Berry S.J., et al, 1986 - Effect of age, castrtation and testosterone replacement on
the development and restoration of canine benign prostatic hyperplasia, Prostate
9:295.
3. Bojrab Joseph M., 1998 - Curent tehniques in small animal surgery, fourth edition,
Williams &Wilkins: 461-511.
4. Harari J., Dupuis J., 1995 - Surgical Treatmens for prostatic disease in dog. Semin
Vet Med Surg 10:43-47.
5. Hardie E.M., 1989 - Peritonities from urogenital conditions, probl Vet Med 1:36.
6. Heradie E.M. et al, 1984 - Complications of prostatic surgery, J Am Anim Hosp
Assoc 20:50.
7. Hargiss A.M., Miller L.M., 1983 - Prostatic carcinoma in the dog, Comp Cont Educ
Pract Vet 5:647.
8. Johnston G.R., 1991 - Diagnostic Imaging of the male canin reproductiv organs:
Methods and limitations, Vet Clin North Am 21:553-590.

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UTILIZAREA UNOR MATERIALE SINTETICE


PENTRU NLOCUIREA LIGAMENTELOR NCRUCIATE
LA CINE
SYNTHETIC MATERIALS USED
FOR DOG CRUCIATE LIGAMENTS REPLACEMENT

A TIMEN, A. MUSTE,
L. OANA, Monica COVACIU

There is no material accepted to substitute in perfect conditions the


cruciate ligaments in dogs.
Different surgical techniques depending on the pacient size and
presence or absence of degenarative joint changes were performed.
Every technique have advantages and desadvantages and thats why
is very difficult to choose the perfect one.
The paper presents the results obtained in using Mersilene and
Nailon tape in cruciate ligaments replacement in dogs.

MATERIAL I METOD

Cercetrile au fost efectuate pe 11 cazuri clinice prezentate la consultaie


cu chioptur de gradul I si II la unul sau ambele membre posterioare.
n funcie de talia animalelor, vrsta i caracterul acut sau cronic al
afeciunii am folosit tehnica operatoare pe care am considerat-o optim. Cazurile
urmrite sunt prezentate n tabel.
Vrsta animalelor a fost cuprins ntre 1 i 3 ani. Examenul radiologic
efectuat cu dubl expunere medio-lateral i cranio-caudal a zonei grasetului a
furnizat date importante referitoare la afeciunile intraarticulare. Mrirea spaiului
articular femuro-tibial sau evidenierea artrofitelor pe condilii femurali au fost
indicii importante pentru diagnosticul afeciunilor ligamentelor ncruciate.
Datorit faptului c aceast afeciune apare la pacieni cu mas corporal
i vrste foarte diferite alegerea tehnicii chirurgicale este un pas foarte important
pentru chirurgul veterinar.
Decizia terapeutic trebuie s in seama de talia cinelui, temperamentul
acestuia, caracterul acut sau cronic al afeciunii, eventualele modificri articulare
i nu n ultimul rnd de cooperarea proprietarului pentru recuperarea animalului.

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Tabelul 1
Animalele luate n studiu care au prezentat ruptura
ligamentelor ncruciate

Greutatea
Nr. Varsta Metoda de
Rasa Sex corporala Diagnostic
Crt. (ani) tratament
(kg)
1. Copoi Banda de
2 M 28 Ruptura LI CR. Dr.
ardelenesc Mersilene
2. Mastino
3 M 60 Ruptura LI CR. Dr. Fir de Nylon
napoletano
3. Pittbull 1 M 30 Ruptura LI CR. Stg. Fir de Nylon
4. Schnauzer
2 F 32 Ruptura LI CR. Dr. Banda de Nylon
urias
5. Banda de
Labrador 3 M 35 Ruptura LI CR. Stg.
Mersilene
6. Terra Nova 3 M 50 Ruptura LI CR. Dr. Banda de Nylon
7. Banda de
Dog German 1 F 45 Ruptura LI CR. Stg.
Mersilene
8. Ciobanesc
2,5 M 30 Ruptura LI CR. Stg. Banda de Nylon
german
9. Caniche 5 M 9 Ruptura LI CR. Dr. Fir de Nylon
10. Cocker spaniel 3 F 9 Ruptura LI CR. Stg. Fir de Nylon
11. Ciobanesc
2 M 28 Ruptura LI CR. Dr. Banda de Nylon
german

REZULTATE I DISCUII

n cazul pacienilor de talie mic n cazul ruperii ligamentului ncruciat


cranial este n general suficient excizia resturilor ligamentare. Aceast manoper
se poate face uor prin artroscopie nefiind necesar intervenia chirurgical
clasic.
Rezultatul unei intervenii chirurgicale corect executate poate s fie
compromis de nesupravegherea atent sau dezinteresul proprietarului n perioada
de convalescen a cinelui.
Nu am semnalat procese septice consecutive actului operator iar
recuperarea funcional a membrului a fost foarte rapid. Astfel la 7-8 zile dup
intervenie a reaprut sprijinul pe vrful degetelor membrului operat pentru ca la 3
sptmni chioptura s dispar complet.
Tensionarea prea puternic a firului poate duce la deirri ale ligamentului
patelar. Pentru a asigura o rezisten i stabilitate mai mare recomandm ca firul
s fie trecut prin ligamentul patelar sub forma literei U, perpendicular pe
traiectul fibrelor acestuia.
n plus n cazul unor defecte minore de conformaie ale scheletului
membrului posterior asociate cu rotaia intern minor a tibiei, prin folosirea
acestei metode se realiniaz ligamentul tibio-rotulian.

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n cazul ruperii firului de nylon, chioptura revine dup o perioad de


restabilire complet a pacientului. Leziunile meniscale trebuie suspectate cnd la
micri succesive de flexie-extensie se percepe un uor pocnet intraarticular.
La unul dintre cazuri n zona crestei tibiale am executat dou orificii la
distan de 5 mm unul de altul. Cele dou fire de nylon le-am trecut prin aceste
orificii sub forma literei U iar apoi n jurul sesamoizilor femurali. Prin acest
procedeu s-a realizat o mai bun stabilitate articular dect n cazul cnd firele
sunt trecute printr-un singur orificiu tibial. Contactul permanent al firelor cu
creasta tibial, n timp poate produce erodarea acestora urmat de ruperea lor.
Tehnicile intracapsulare de refacere a ligamentului ncruciat anterior rupt
constau n nlocuirea ligamentului rupt cu o gref provenind din materiale
sintetice (Mersilene, Nylon).
La alegerea tehnicii trebuie s se in cont de greutatea corporal a
animalului i de caracterul acut sau cronic al leziunii, prezena sau absena
fenomenelor degenerative articulare i vrsta animalului.
Chiar dac din punct de vedere operator tehnicile sunt apropiate , rolul
deosebit de important al operatorului este acela de a alege metoda terapeutic
optim pentru fiecare caz n parte.
Ca material de substituire a ligamentului am folosit banda de Mersilene i
banda de Nylon.
Banda de Mersilene utilizat este folosit n chirurgia abdominal uman
pentru uteropexie. Limea acesteia este de 4 mm fiind ambalat steril de firma
productoare.
Banda a fost introdus iniial prin orificiul de la nivelul crestei tibiale unde
captul acesteia a fost nnodat. n continuare banda strbate tunelul tibial n sens
ventro-dorsal iar apoi cel femural, captul ajungnd n zona condilului lateral
femural.
Tensionarea benzii s-a realizat meninnd articulaia grasetului n extensie
iar captul proximal al acesteia l-am suturat n puncte separate cu fir neresorbabil,
Nylon, la esuturile moi din zona periarticular lateral. Dup verificarea
stabilitii articulare, nchiderea plgii operatorii se realizeaz dup tehnica
uzual.
n primele 5-6 zile postoperator dup dispariia edemului operator
vindecarea a decurs normal. Sprijinul pe membrul operat nu s-a reluat ns nici
dup 21 zile de la intervenie la unul dintre cazurile la care am aplicat aceasta
tehnica.
La 4 sptmni de la prima intervenie, pacientul a fost supus unei noi
intervenii chirurgicale prin care am ndeprtat banda de Mersilene i am nlocuit-
o cu un ligament obinut din fascia lata.
Vindecarea a decurs normal, fr complicaii i la 30 zile de la a doua
intervenie sprijinul pe membrul operat a fost reluat. Dup 45 de zile chioptura
a disprut complet.

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Faptul c dup eliminarea benzii de Mersilene i nlocuirea acesteia cu


fascia lata, fenomenele supurative au disprut ne-au condus la concluzia c
materialul din care era fabricat ligamentul artificial a generat in acest caz reacia
de respingere a grefei.
n alte 4 cazuri am folosit banda de Nylon pentru nlocuirea ligamentului
ncruciat cranial rupt .Fixarea benzii am realizat-o n dou moduri. n doua cazuri
traiectul benzii a fost identic cu cel descris la banda de Mersilene iar n 2 cazuri
capetele benzii au fost nnodate pe faa lateral a crestei tibiale. Prin folosirea
acestui material sintetic nu am sesizat reacii de respingere sau ruperea
implanturilor.
Procesele de cicatrizare au decurs normal iar reluarea complet a sprijinului
pe membrul operat s-a realizat n medie dup 30 zile.
Problema gsirii unui material perfect pentru nlocuirea ligamentelor
ncruciate rupte a preocupat i continu s preocupe chirurgii veterinari. Multe
materiale incluznd Teflon , Nylon sau Dexon au fost ncercate dar nici unul nu a
fost considerat perfect.
Acesta ar trebui s aib o rezisten mare, o uoar elasticitate i s suporte
fore de ntindere n interiorul articulaiei timp de cteva luni, totodat nefiind
iritant. De aceea cercetrile pentru gsirea materialului ideal continu..
n cercetrile ntreprinse am ncercat aplicarea benzii sub forma unei anse
astfel nct cele dou capete se reunesc i se nnoad pe faa lateral a crestei
tibiale. Prin folosirea acestei tehnici am observat c reluarea sprijinului pe
membrul operat este mai rapid, dect n cazurile utilizrii metodei clasice.
Probabil acest fapt se datoreaz elasticitii mai mari a ligamentului aplicat astfel
i eliminrii traciunii pe esuturile moi n zona suturii pe faa lateral a condilului
femural lateral.
La cazurile operate i urmrite chiar pe parcursul a 2 ani nu am observat
apariia fenomenelor de respingere a grefei artificiale din Nylon sau erodarea
urmat de ruperea acesteia.
n cazul folosirii benzilor de Nylon pentru nlocuirea ligamentului
ncruciat cranial trebuie avut n vedere masa corporal a pacienilor. Adaptarea
limii benzii n funcie de talia fiecrui pacient este una din etapele cele mai
importante ale interveniei. O band cu lime mic de 3 mm nu va putea suplini
funcia unui ligament ncruciat la un cine de 50-60 kg i se va rupe.
n acelai timp o band cu lime mare de 6-8 mm pentru a putea fi aplicat
la un cine de talie mic (sub10 kg), necesit efectuarea unor orificii n epifiza
distal femural i cea proximal tibial cu diametrul de cel puin 5-6 mm.
Acestea vor micora rezistena osoas n aceste zone mrind riscul unor fracturi
epifizare.

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CONCLUZII

Ruptura ligamentului ncruciat cranial poate s fie ntlnit la cini


indiferent de vrsta i rasa acestora. Totui sunt predispui cinii aparinnd
raselor cu greutate corporal mare.
Afeciunea debuteaz cu chioptur de gradul I care se agraveaz treptat
ajungnd la evitarea efecturii sprijinului pe membrul afectat.
Diagnosticarea precoce limiteaz fenomenele degenerative datorate
instabilitii articulare marcante.
Conduita terapeutic trebuie s in seama de talia, vrsta i de prezena sau
absena unor afeciuni asociate ca luxaia rotulian, leziuni ale meniscurilor
articulare sau fenomene degenerative articulare secundare.
Dei nc nu s-a descoperit un material sintetic perfect pentru nlocuirea
ligamentului ncruciat, prin folosirea benzii de Nylon la rasela de talie corporala
mare si a firului de Nylon la rasele mici am obinut rezultate foarte bune.

BIBLIOGRAFIE

1. Arnoczky S.P., 1990 - Cranial cruciate ligament repair. In: Bojrab MJ, ed. Current
Techniques in small animal surgery. 3rd ed. Philadelphia: Lea&Febiger, 708.
2. Arnoczky S.P., 1981 - Surgery of the stifle: The cruciate ligaments. Compendium on
Continuing Education for the Small Anim Pract 2:106.
3. Butler D.L., et al., 1989 - Mechanical properties of primate vascularized vs.
nonvascularized patellar tendon graft: changes over time. J Orthop Res; 7: 68-
79.
4. Butler H.C., 1964 - Teflon as a prosthetic ligament of ruptured anterior cruciate
ligaments. Am J Vet Res 25:55, 1964.
5. Piermattei D.L., 1993 - An atlas of surgical approaches to the bones and joints of the
dog and cat. 3rd ed. Philadelphia: WB Saunders, 114.

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EPIDEMIOLOGIA OTITEI EXTERNE LA CARNIVORE


EPIDEMIOLOGY OF OTITIS EXTERNA AT CARNIVORES

Roxana TOPAL, I. BURTAN,


M. FNTNARU, S. CIOBANU.,
L.C. BURTAN, M. TOPAL
U.S.A.M.V. Iai

Otitis externa is the most common disease of the canine and feline
ear canal, which involves an acute or cronic inflammation of the epithelium
of the external auditory meatus.
The goal of this study was to determinate the prevalence rate of otitis
externa and the influence of species, breed, age, sex and climatic variation.
The researche was performed on 356 canine and 83 feline which were
atended in Clinics of Veterinary Medicine Faculty and private veterinary
clinics.
Dogs and cats of all breeds and ages may be affected, but some
groups are at higher risk:Cocker, Labrador, Setter, German Shepherd had
the highest incidence. Dogs between 5-8 years of age and cats under 1 year
are commonly affected. There is no apparent sex predisposition to otitis
externa in either species.

Key words: otitis, incidence, prevalence, canines, felines

Otita extern, denumit n literatura de specialitate i catar auricular,


reprezint inflamaia acut sau cronic a epiteliului conductului auditiv extern,
uneori i a pavilionului urechii (1, 3, 6, 8). Este o afeciune cu etiologie
polifactorial i patogenez neelucidat pe deplin.
Baba, 1981, precizeaz c otita extern afecteaz cini i pisici de toate
rasele i vrstele, cu inciden mai mare la unele rase datorit unor particulariti
anatomice si fiziologice. Examenele histologice comparative ale conductului
auditiv, la diferite rase de cini, indic diferene privind numrul i distribuia
glandelor ceruminoase, explicnd astfel predispoziia unor rase la otit (1, 6).
Studiul epidemiologic al otitei externe se impune, att pentru elucidarea
etiopatogenezei, ct i pentru stabilirea corelaiei ntre frecvena otitei externe i
anumii factori, printre care am inclus: rasa, vrsta, sexul i variaiile climatice.

MATERIAL I METOD

Materialul de studiu a fost reprezentat de cazuistica oferit de Clinicele din


cadrul Facultii de Medicin Veterinar Iai i Cabinete medicale veterinare.
Pentru stabilirea frecvenei otitei externe la carnivore, studiile au fost efectuate pe
perioada 2002 2005 inclusiv.

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REZULTATE I DISCUII

Parametrii de baz ai studiului epidemiologic sunt morbiditatea i


mortalitatea. Morbiditatea se refer la stabilirea numrului total de cazuri de otit
nregistrate la carnivore, n perioada luat n studiu. n cadrul morbiditii s-a
operat cu doi indicatori: incidena i prevalena. Incidena se refer la numrul de
cazuri noi nregistrate ntr-un interval de timp, raportat la totalul populaiei
studiate. Prevalena const n stabilirea numrului total de cazuri nregistrate,
respectiv noi i vechi, la un anumit moment, conform studiului la sfritul
perioadei de patru ani (7).
Analiza noastr nu opereaz cu termeni adecvai de morbiditate, inciden,
prevalen datorit lipsei unor populaii mari de referin, respectiv toate
carnivorele de pe un anumit areal, ci cu termenul de frecven relativ, ntruct nu
toate animalele bolnave sunt prezentate la medicul veterinar, astfel nct doar cele
care beneficiaz de asisten sunt supuse anchetei epidemiologice.
Evidenele unitilor care au furnizat datele ne permit stabilirea frecvenei
relative a otitei la carnivore, datele fiind prezentate n tabelul 1.
Tabel 1
Frecvena relativ a otitei externe la carnivore
n perioada 2002-2005

Animale Din care cu otit Masculi Femele


Specia
consultate Nr. % Nr. % Nr. %
Cini 4275 356 8,3 185 52 171 48
Pisici 3864 83 2,15 37 44 46 56

Din analiza datelor din tabel se observ c otita extern are o frecven
relativ mai mare la cini, reprezentnd 8,3% din cazuistic, fa de pisici la care
frecvena relativ este de 2,15%. Frecvena relativ sczut a otitei externe la
pisici se datoreaz poziiei erecte a pavilionului urechii i lipsei prului din
conductul auditiv extern, care permit ventilaia i drenajul (1, 2, 6).
Analiza datelor din tabel evideniaz, de asemenea, faptul c nu exist o
corelaie clar ntre sexul animalului i frecvena relativ a otitei externe.
Afirmaia se bazeaz pe faptul c procentele oferite de cele dou sexe sunt
apropiate sau uor crescute n favoarea masculilor la canine sau femelelor la
feline.
n cadrul studiului epidemiologic am evideniat rolul speciei, rasei, vrstei
i sexului n apariia otitei externe la carnivore, precum i repartizarea lunar a
cazuisticii, datele fiind prezentate n tabelele 2, 3, 4, 5 i 6.
Analiza datelor din tabel evideniaz rolul rasei n apariia otitei externe la
cine, constatndu-se c otita extern apare mai frecvent la unele rase datorit
unor particulariti anatomice i fiziologice. Astfel, 73,2% din cazuri apar la cinii
cu urechile atrnnde i doar 26,8% la rasele cu urechile erecte. Dintre cei cu

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urechile atarnnde o frecvena relativ crescut s-a nregistrat la rasele Cocker


(26%), Labrador (18%) i Setter (13,3%). La rasele cu urechile erecte cea mai
mare frecven se nregistreaz la Ciobnescul german (14%).
Tabelul 2
Frecvena relativ a otitei externe la cini n raport cu rasa

Cu urechile atrnnde Nr. %


Brac 20 5,8
Caniche 13 3,7
Cocker 92 26
Labrador 64 18
Pechinez 12 3,2
Setter 47 13,3
Ras comun 12 3,2
Total 260 73,2
Cu urechile erecte Nr. %
Akita 7 2,1
Boxer 7 2,1
Ciobnesc german 51 14
Doberman 4 1,1
Dog Argentinian 6 1,6
Pit-Bull 6 1,6
Rottweiler 11 3,2
Ras comun 4 3,1
Total 96 26,8
TOTAL GENERAL 356 100

Tabelul 3
Frecvena relativ a otitei la pisici n raport cu rasa

Rasa Nr. %
Birmanez 26 31,3
Persan 21 25,4
Siamez 22 26,5
Ras comun 14 16,8
Total 83 100

La pisici, frecvena relativ a otitei externe n raport cu rasa, nregistreaz


procente asemntoare ca valoare. Mai predispuse la otit sunt pisicile din rasa
Birmanez, la care s-au nregistrat 26 de cazuri, reprezentnd 31,3%, cele din rasa
comun nscriind procentul cel mai mic, respectiv 16,8%.
n ceea ce privete corelaia dintre vrst i frecvena relativ a otitei
externe, observm c exist diferene ntre cele dou specii de carnivore.

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Tabelul 4
Frecvena relativ a otitei externe la cine n raport cu vrsta

Vrst Nr %
sub 1 an 23 6,4
1 2 ani 60 17
2 5 ani 109 30,6
5 8 ani 121 33,8
peste 8 ani 43 12,2
Total 356 100

Din analiza datelor din tabelul 4 remarcm faptul c otita extern afecteaz
cini de toate vrstele. S-au nregistrat 23 de cazuri (6,4%) la cinii sub 1 an, apoi
se observ o cretere progresiv a numrului de cazuri n raport cu vrsta: 60 de
cazuri (17%) la cei cu vrsta cuprins ntre 1-2 ani, 109 cazuri (30,6) la cinii de
2-5 ani i 121 de cazuri (33,8%) la cinii cu vrsta cuprins ntre 5 i 8 ani.
Frecvena relativ a otitei externe scade la cinii mai mari de 8 ani nregistrndu-
se 43 de cazuri (12,2%).
Creterea incidenei este recunoscut de majoritatea autorilor ca fiind
plasat n jurul vrstei de 5-8 ani, deoarece n acest interval apar tulburrile de
cheratinizare i de hipersensibilizare (1, 2, 8).
Tabelul 5
Frecvena relativ a otitei externe la pisic n raport cu vrsta

Vrst Nr %
Sub 1 an 29 35
1 2 ani 19 23
2 5 ani 23 28
Peste 5 ani 12 14
Total 83 100

Analiznd datele din tabelul 5 observm c otita extern afecteaz cel mai
frecvent pisicile tinere, 35% din cazuri nregistrndu-se la pisicile sub un an.
Frecven crescut ntlnim i la pisicile cu vrsta cuprins ntre 2 i 5 ani (28%)
deoarece dup vrsta de doi ani exist o posibilitate mai mare de apariie a otitei
parazitare (5).
n tabelul 6 sunt centralizate datele prezentrii la consultaie a celor 356 de
cini i 83 de pisici cu otit extern.
Analiznd datele din tabel remarcm o concentrare a cazuisticii n perioada
de var: iunie 14,6% la cine, respectiv 13,2% la pisic, iulie 15,7% la cine i
15,6% la pisic, august 13,8% la cine, iar la pisic 14,4%. Aceste procente ne
orienteaz spre o evoluie sezonier a otitei externe la carnivore, influenat de
condiiile de mediu. Creterea temperaturii i a umiditii mediului extern
determin o cretere a frecvenei relative a otitei externe la carnivore (1,6). De

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asemenea, exist i o probabilitate mai mare de apariie a otitei externe datorat


corpilor strini (ariste), dar i a otitei parazitare (2, 6).
Tabelul 6
Repartizarea lunar a cazuisticii

Prezentarea la consultaie n luna


Specia
I II III IV V VI VII VIII IX X XI XI
Nr. 9 13 17 29 38 52 56 49 31 27 20 15
Cine
% 2,5 3,6 4,8 8,1 10,6 14,6 15,7 13,8 8,7 7,6 5,8 4,2
Nr. 2 2 4 7 9 11 13 12 10 6 4 3
Pisic
% 2,4 2,4 4,8 8,4 10,8 13,2 15,6 14,4 12,4 7,2 4,8 3,6

CONCLUZII

Epidemiologia otitei externe la carnivore reprezint un studiu complex care


trebuie corelat cu o serie de factori: specie, ras, vrst, sex, variaii climatice.
Incidena i prevalena otitei externe sunt indicatori orientativi deoarece nu
toi indivizii speciei, de pe un anumit areal, au fost prezentai la medicul veterinar.
Frecvena relativ a otitei externe este mai mare la canine, n comparaie cu
felinele.
Rasa influeneaz frecvena relativ a otitei la canine, procente mari fiind
nregistrate la cinii cu urechile atrnnde (73,2%).
Cele mai multe cazuri au fost nregistrate la caninele cu vrsta cuprins
ntre 5-8 ani i la felinele sub 1 an.
Nu exist o corelaie evident ntre sex i frecvena relativ a otitei externe
la cini i pisici.
Otita extern la carnivore are o evoluie sezonier, nregistrndu-se o
frecven relativ crescut n lunile de var.

BIBLIOGRAFIE

1. August, J.R., 1986 Diseases of the ear canal In The Complete Manual Of Ear
Care, Lawrenceville, New Jersey, Veterinary System Co., Inc., 37-51
2. Baba, E., 1981- Incidence of otitis externa in dogs and cats in Japan, Vet. Rec., 108-
393
3. Burtan I., Eugenia Burtan, Burtan L.C., Mihai M., 1999 Contribuii la studiul otitei
externe la pisic, Lucr.t. vol.42 seria M.V., Ed. I. I. de la Brad Iai, 82-86
4. Burtan, I., 2000 Chirurgie regional veterinar, Ed. Ion Ionescu de la Brad, Iai
5. Cosoroab, I., 2000 Parazitologie veterinar, Ed. Mirton, Tim.
6. Griffin, C.E., 1981 Otitis externa, Comp. Contin. Ed. Pract. Vet., nr. 3, 741-750
7. Savu, Gh., 2001 - Epidemiologie veterinar general, Ed. I.I. de la Brad Iai
8. Solcan, Gh., Miron L., Mitrea, I.L., Carmen Solcan, 2003 Dermatopatologia
animalelor de companie, Ed. Ion Ionescu de la Brad, Iai.

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OBSERVAII PRIVIND INCUBAIA OLOR DE STRU


M. BADEA

Lucrarea trece n revist dinamica procesului produciei de ou,


incubaia i ecloziunea ntr-o ferm de strui i perspectivele de
mbuntire a acestor indicatori.
Studiul a luat n considerare factorii care pot influena indicatorii de
producie ntr-o ferm privat de cretere a struilor, prin cunoaterea
elementelor ce caracterizeaz evoluia, comportamentul i viaa struilor n
vederea unei creteri eficiente.

Struul face parte din clasa Aves, subclasa Palaeognathae, familia


Struthionidae, reprezentat prin singura specie actuala Struthio camelus, care a
dat natere la diferite rase geografice i populeaz natural regiunile de step i
semideert din Asia i Africa. (Deeming, D., 1988, Bonaiu, I., 1996, Vacaru
Opri, I., 2002)
Creterea struilor n fermele de producie mai mari sau mai mici a luat un
avans deosebit n ultimii ani datorit importanei mari i valorii deosebite a
tuturor produselor lor ct i a productivitii lor. (Smith, W.A., 1995)
Observaiile noastre au urmrit dinamica procesului de incubaie i
ecloziune ntr-o ferm privat de cretere a struilor.

MATERIAL I METOD

Cercetrile au fost efectuate pe 8 familii de strui compuse din 24 de indivizi


(16 femele i 8 masculi). Pe parcursul a 5 ani s-au fcut observaii comparative a
mai multor parametri ai incubaiei astfel:
- nr. total de ou;
- ou cu defecte (ou sparte, cu coaja moale, coaja poroas, ou cu
depuneri neregulate de calciu);
- ou puse la incubat;
- ou eliminate la primul i al II lea miraj;
- ou cu pui mori;
- nr. pui eclozionai.

REZULTATE I DISCUII

Avndu-se n vedere faptul c ferma a fost populat la data de 02.07.2001


rezultatele obinute n acest an nu sunt foarte concludente. Astfel s-au obinut 56
de ou, 6 prezentau defecte, majoritatea nefiind fertile. S-au pus la incubat 13 ou
(24%) din acestea 4 ou (31%) eliminndu-se la primul miraj. n total, n 2001 au
eclozionat 9 pui (69%).

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n anul 2002 producia de ou a crescut la 196 din care; 8 ou (4%)


prezentau defecte. S-au pus la incubat 146 de ou (75%), eliminndu-se la
examenul ovoscopic 24 de ou (16%). Datorit furajului slab calitativ,
nembuntit n vitamine i oligoelemente 24 de ou (16%) aveau puii mori n
diferite stadii de cretere. Astfel n 2002 au eclozionat 88 pui (60%) din cele 147
ou puse la incubat.
Tabelul 1
Rezultatul incubaiei pe ani

Ou la Eliminate la Pui mori Pui Ou cu


OU incubat ovoscopie n ou eclozionai defecte
Anul
TOTAL
nr. % nr. % nr. % nr. % nr. %
2001 56 13 24 4 31 - - 9 69 6 11
2002 196 147 75 35 24 24 16 88 60 8 4
2003 339 209 62 98 47 19 9 92 44 14 4
2004 391 244 62 133 55 27 11 84 34 4 1
2005 307 58 19 38 66 4 7 16 28 5 2
Total 1.289 671 52 308 46 74 11 289 43 37 3

n 2003 ncepnd cu jumtatea lunii februarie furajul s-a mbuntit cu


concentrat LIKRA bogat n vitamine i oligoelemente, furaj special pentru
stimularea ouatului la strui. Astfel n 2003 s-a obinut o producie de ou de 339
buc, 14 ou prezentnd defecte (4%). S-au pus la incubat 209 ou (62%),
eliminndu-se la examenul ovoscopic 98 de ou (47%). Din cele rmase 19 ou
(9%) aveau puii mori n diferite stadii, reuind s eclozioneze la finele anului un
numr de 92 pui (44%), aproape jumtate din totalul olor puse la incubat.
Datorit unor afeciuni cronice 2 femele bune outoare au murit n iarna
anului 2003 fapt care a influenat negativ producia de ou din anul 2004.
n anul 2004 s-au obinut 391 ou din care 4 (1%) prezentau defecte
minore. S-au pus la incubat 244 ou (62%) eliminnd dup primul miraj 133 de
ou (55%) i 27 de ou (11%) prezentau puii mori n diferite stadii. Astfel, n
anul 2004 au eclozionat 84 pui (34%) din totalul oulor puse la incubat.
n anul 2005 datorit unor probleme organizatorice i tehnice (defeciuni
ale sistemului de incubaie) valorile acestor parametri nu au fost mai sczute.
Astfel, s-a obinut o producie de ou de 307 buc din care 5 ou (2%)
prezentau defecte de structur. S-au pus la incubat 58 ou (19%) eliminndu-se la
examenul ovoscopic 38 ou (66%) iar 4 ou (7%) aveau embrionii mori n ou.
Au eclozionat 16 pui, doar 28% din totalul olor puse la incubat.
Fcnd un studiu centralizat al incubaiei pe toat perioada de 5 ani
constatm urmtorii indicatori de producie :
- - total ou obinute = 1.289 buc ;
- - din care ou cu defecte = 37 buc (3%) ;
- - ou puse la incubat = 671 buc (52%) ;

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- - ou eliminate la examenul ovoscopic = 308 buc (46%) ;


- - ou cu puii mori n ou = 74 buc (11%)
- - pui eclozionai = 289 cap. (43%).

CONCLUZII

Din studiile efectuate pentru evaluarea indicatorilor de producie i


incubaie n perioada de cercetare constatm urmtoarele :
1. nceputul i continuarea perioadei de ouat este dependent de factori de
mediu: condiiile climatice existente, factori de stres, calitatea furajului
(vitaminele i oligoelementele necesare acestei perioade). Lumina influeneaz
ouatul i stimularea glandei pituitare de a produce hormoni, maturarea ovulei i
stimularea produciei de sperm la mascul. Corelaia pozitiv ntre lumin i
temperatur favorizeaz fecunditatea i eclozionabilitatea.
2. Alimentaia complet, dar nu excesiv, evitnd ngrarea acestora
(punnd la dispoziie sfrmtur de scoici, iarb, fn i combinatele s aib
valori optime de aminoacizi, vitamine i oligoelemente) influeneaz pozitiv
fecunditatea.
3. Formarea familiilor selectnd indivizi de mare productivitate (F113 = 67
ou, F140 = 67 ou, F148 = 53 ou) cu masculi buni reproductori este
important n exploatare.
4. Procentul total de fecunditate obinut pe aceast perioad este de 52%
5. Procentul de ecloziune (43%) este influenat de unii parametri tehnici de
incubaie (t umed 24,8 26,2 : t uscat 36,2 36,6).
6. Din observaiile efectuate de noi, procentul de ou cu deformaii se
ncadreaz ntre 1 i 4.

BIBLIOGRAFIE

1. Blescu, M., 1999 Creterea struilor.


2. Bonaiu, F., 1996 Farmecul columbofiliei. Ed. POPAS ART Timioara.
3. Deeming, D.C., 1988 Ostrich Chick Rearing. United Kingdom.
4. Smith, W.A., 1995 Feeding and Feed Management. Rep. South Africa.
5. Vacaru, Opri, I., 2002 Tratat de avicultur. Ed. Ceres Bucureti.

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OBSERVAII PRIVIND DINAMICA RETENIILOR


NVELITORILOR FETALE I INFLUENA ACESTORA
ASUPRA UNOR PARAMETRI DE REPRODUCIE
OBSERVATIONS CONCERNING THE DYNAMIC OF FETAL MEMBRANES
RETENTIONS AND THEIR INFLUENCE ON SOME REPRODUCTION PARAMETERS

M. CANTEMIR

The fetal membranes retentions and endometritis are frequent


affections of milk cows bred in intensive sistem.
The presence of these affections is influenced by milk production, the
monthly dynamic of retentions and endometritis and by the animals age.
An increasment frequence of fetal retentions rep until 36, 25% is
observed at productions over 5000 l on normal lactation. In the cold season
the most frequent cases : 47,5% are registred unlike 7,5% during the
summer season.

La vacile pentru lapte crescute n sistem intensiv, reteniile nvelitorilor


fetale i endometritele sunt afeciuni ce apar destul de frecvent (Boitor, I., 1987;
Gluhovschi, N., 1971; Gluhovschi, N., Drugociu, Gh., Seiciu, Fl., Dumitrescu, I.,
1972; Grunert, E., 1978; Hricu Valentina, 1982; Runceanu, L., 1995; Scheideger,
G.A., Melendez, R.P., 1994; Seiciu, Fl., Drugociu, Gh., Boitor, I., 1989; Broiu, I.,
Seiciu, Fl., 2004; Cernescu, H., 2004).
Prezena acestor afeciuni este influenat de producia de lapte, de
dinamica lunar a reteniilor i a endometritelor, precum i de vrsta animalelor.
Se constat c frecvena reteniilor fetale crete pn la 36,25%, la
produciile de peste 5000l pe lactaie normal. Cele mai frecvente cazuri se
nregistreaz n anotimpul rece 47,5%, fa de numai 7,5% n anotimpul de var.

MATERIAL I METOD DE LUCRU

Observaiile noastre au fost efectuate n cadrul unei ferme de cretere


industrial a vacilor pentru lapte.
Diagnosticul cazurilor cu retenia anexelor fetale s-a efectuat clinic lundu-
se n considerare mai multe aspecte:
- momentul parturiiei
- perioada eliminrii anexelor fetale
- eliminarea total sau parial a acestora
n acest sens s-a considerat fa de media de 12 ore (eliminarea normal a
placentei la vac), c supravegherea parturientei se efectueaz pn la 24 ore
postpartum. La acest termen intervenia medicului veterinar este obligatorie.

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REZULTATELE OBINUTE I INTERPRETAREA LOR

Procentul mare de retenii ale anexelor fetale, la produciile de peste 5000


litri lapte anual, poate fi explicat prin factorul de stres pe care-l reprezint nivelul
ridicat al produciei, acionnd prin sistemul feed-back negativ suprarenal-
hipotalamus-adenohipofiz, ca un factor al centrilor responsabili de producia
lactat. Totodat, aceti factori acioneaz negativ asupra dinamicii uterine
postpartum, contraciile uterine fiind unul din cei mai importani factori care
asigur eliminarea nvelitorilor fetale.

Fig. 1. Frecvena endometritelor n funcie de producia de lapte.

Din analiza graficului din fig. 1 constatm influena pe care o are producia
de lapte asupra apariiei endometritelor la vaci. Astfel, n cazul vacilor cu
producii de 2500-3000 litri lapte pe an , cazurile de endometrite sunt rare, de
9,2-10%. n cazul vacilor cu producii mari de lapte , de 4000-5000 litri pe an,
numrul cazurilor de endometrit crete considerabil, atingnd valori de 25-
37,4%.
Datorit factorilor care condiioneaz creterea ponderii reteniei anexelor
fetale n funcie de producia de lapte, observaiile noastre reliefeaz i o cretere
a infeciilor endometriale puerperale.
Cercetrile noastre arat c nu numai nivelul produciei de lapte
influeneaz apariia reteniilor nvelitorilor fetale i a endometritelor, ci i unii
factori de mediu.
Analiza datelor din tabelul 1 arat c cele mai frecvente cazuri se
nregistreaz n anotimpul rece 47,5%, fa de numai 7,5% n anotimpul de var i
12,5% n anotimpul de toamn. De asemenea , reteniile nvelitorilor fetale sunt
diagnosticate cu o frecven mare n anotimpul de primvar (32,5%).
Ponderea ridicat a reteniilor nvelitorilor fetale n cele dou anotimpuri,
iarna i primvara, se datoreaz unor carene alimentare, unor condiii zooigienice

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necorespunztoare i stabulaiei permanente. Odat cu schimbarea alimentaiei


(furaje verzi), asociat cu micarea la pune, tonusul general al organismului se
revigoreaz, dinamica uterin este mai bun i n consecin frecvena reteniilor
nvelitorilor fetale scade foarte mult.
Tabelul 1
Frecvena reteniilor nvelitorilor fetale pe luni i anotimpuri

Luna I II III IV V VI VII VIII IX X XI XII


Nr. cazuri 4 8 6 3 4 2 1 2 3 2 2 2
% fa de
nr. total de 10 20 15 7,5 10 5 2,5 5 7,5 5 5 7,5
cazuri
Anotimp Primvar Var Toamn Iarn
Nr. cazuri 13 3 5 19
%fa de
nr. total de 32,5 7,5 12,5 47,5
cazuri

Frecvena reteniilor nvelitorilor fetale pe luni ale anului este prezentat n


tabelul 1, unde se constat c n lunile februarie i martie se ntlnesc cele mai
sczute cazuri, 20% respectiv 15%, n timp ce n luna iulie am ntlnit un singur
caz de retenie a nvelitorilor fetale, reprezentnd 2,2% din numrul total de
cazuri.
n cazul apariiei reteniilor, constatm i o cretere a I.G. (indice de
gestaie), numrul de nsmnri artificiale pentru obinerea unei gestaii
ajungnd pn la 3,8, de unde concluzia c productivitatea i rentabilitatea sunt
mult diminuate.

CONCLUZII

1. Reteniile nvelitorilor fetale sunt influienate de producia de lapte. La


vacile cu producii mari de lapte, frecvena acestor afeciuni crete foarte mult,
ajungnd pn la 36,25%, n timp ce la o producie de 2500 litri pe an frecvena
reteniilor nvelitorilor fetale scade considerabil ajungnd pn la 8,45%.
2. Reteniile nvelitorilor fetale i endometritele puerperale apar cu o
frecven mai mare iarna 47,5%, respectiv 40% i primvara 32.5%, respectiv
33.3% i mai rar vara 7,5%, respectiv11,1% i toamna 12,5% respectiv 15,5%.
3. Ponderea reteniilor nvelitorilor fetale n cele dou anotimpuri, iarna i
primvara, se datoreaz unor carene alimentare, unor condiii zooigienice
necorespunztoare i stabulaiei prelungite.
4. Frecvena reteniilor nvelitorilor fetale n lunile februarie i martie este
mult mai crescut ntlnindu-se n 20% din cazuri n luna februarie i 15% din
cazuri n luna martie, n timp ce n luna iulie s-a ntlnit un singur caz de retenie
a nvelitorilor fetale reprezentnd 2,2% din numrul total de cazuri.

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5. Analiznd variaiile principalilor indici de reproducie concluzionm


faptul c n cazul vacilor cu o retenie a anexelor fetale, S.P. (service-period)
ajunge la o medie de 115,3 zile, constatndu-se totodat o prelungire a C.I.
(calving-interval), care are o valoare medie de 410,2 zile.

BIBLIOGRAFIE

1. Broiu, A., Seiciu, Fl., 2004 Tratat de reproducie la animale. Ed. All. Buc.
2. Boitor, I., 1987 Diagnosticul i tratarea bolilor obstetrico-ginecologice i ale glandei
mamare la bovine, vol. I i II. Ed. Dacia, Cluj-Napoca.
3. Cernescu, H., 2004 Ginecologie veterinar. Ed. Mirton. Timioara.
4. Gluhovshi, N., 1971 Reproducia animalelor domestice, vol. II, Ed. Ceres,
Bucureti.
5. Gluhovschi, N., Drugociu, Gh., Seiciu, Fl., Dumitrescu, I., 1972 Biologia i
patologia reproduciei, Ed. Didactic i Pedagogic, Bucureti.
6. Grunert, E., 1978 Etiology of Retained Bovine Placenta Tenogenology, Curent
Therapy.
7. Runceanu, L., 1995 Reproducia i patologia reproducie, U.A.M.V., Iai.
8. Seiciu, Fl., Drugociu, Gh., Boitor, I., 1989 Reproducia patologic la animalele de
ferm, vol. II, Ed. Ceres, Bucureti.
9. Scheiger, G.A., Melendez, R.P., 1994 Retainde fetal membranes and other
puerperal reproductive disorders their effect on postpartum fertility in Holstein
catlle, Veterinary Bulletin.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

CERCETRI PRIVIND APRECIEREA VALORILOR


REZISTENEI SPERMATOZOIZILOR DE VIER

SEARCHES ABOUT BOAR SPERMATOZOA VALUES OF RESISTANCE

t.G. CIORNEI, L. RUNCEANU


U.S.A.M.V. Iai
G. AGAPE
S.C. Suinprod Roman S.A.

The spermatozoides resistance at the noxious action of a 1% NaCl


electrolites solution it is confered by the seminal cell lipoproteic membrane.
If this lipoproteic membrane is not enough structuraly and functionaly
consolidated, the spermatozoids will have a less longiviability and
fecundity, because of the aggression done by Na+ and Cl- ions.The
literature of specialty does not say which are the limits of variations for the
spermatozoids resistance; its only stating that the minimal limit must be
1000. Thats way this work want to examine this parameter relating him to
the temperature of examination and preservation. Taking in observation a
number of 23 boars and examining the spermatozoids resistance of the
fresh, the diluted and the preserved sperm, shows that the parameters are
much higher than those mentioned in the literature of specialty.

Cuvinte cheie: vier, spermatozoizi, rezisten

Rezistena spermatozoizilor este dat de proprietatea acestora de a rezista la


aciunea nociv a unui toxic. Soluia de clorur de sodiu, concentraie 1% este
folosit n acest scop. Rezistena la aciunea nociv a electroliilor din soluia de
NaCl 1% este conferit de membrana lipoproteic a celulei seminale [6, 8].
Celulele seminale o dobndesc in epididim, in cursul procesului de maturare, ce
dureaz la vier in medie 8 zile [1, 4, 5, 7, 8]. Dac membrana lipoproteic este
insuficient consolidat structural i funcional, spermatozoizii sub agresiunea
ionilor de Na+ i Cl-, vor fi intoxicai de un volum mic din aceast soluie i i
vor pierde proprietatea de micare. Din contr, spermatozoizii care au membran
lipoproteic integr, vor avea capacitatea de a rezista la un volum mai mare din
soluia electrolitic, n acest caz rezistena fiind mai ridicat [6, 8].
Unii autori afirm c longiviabilitatea i fecunditatea spermatozoizilor este
cu att mai ridicat cu ct spermatozoizii i menin mobilitatea ntr-un volum mai
mare de soluie NaCl 1% [6, 8].
Rezistena se exprim prin numrul de ml de soluie NaCl 1% care opresc
micarea de naintare (rectilinie) a spermatozoizilor ce se gsesc ntr-un ml de
material seminal [2, 6, 8]

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Valoarea rezistenei unui material seminal de vier, pentru a putea fi folosit


la nsmnri artificiale, trebuie s aib minim 1000 [2, 3, 6, 8, 9].
Literatura de specialitate nu precizeaz limitele de variaie ale acestui
parametru, ci doar valoarea minim; fapt ce constituie scopul acestei lucrri.

MATERIAL I METOD

Cercetrile s-au efectuat pe un numr de 23 vieri, clinic sntoi, de rase,


linii i vrste diferite. Vierii sunt crescui n condiiile corespunztoare ale
complexului industrial S.C. SUINPROD ROMAN S.A.
Programul individual de recoltri a variat de la 3 la 10 zile de pauz. Dup
recoltare materialul seminal este supus examenelor generale i speciale.
Rezistena spermatozoizilor se determin la microscop pe sperma brut i / sau
diluat.
ntr-un pahar Berzelius se depun cu ajutorul unei pipete Sahli un volum de
0,02 ml sperm, peste acesta se adaug 10 ml solutie NaCl 1%. Se
omogenizeaz uor, se ia cu o baget o picpur de amestec i se examineaz
la microscop n pictur liber. Dac se ntlnesc spermatozoizi cu micri
rectilinii se continu titrarea adugndu-se cte 10 ml soluie, pn cnd
micrile rectilinii dispar. Fiecare titrare a fost urmat de examinarea la microscop
a 2-3 picturi din paharul de amestec. n cmpul microscopic s-au observat,
spermatozoizi cu diferite tipuri de micri: anormale - micri n manej, micri
retrograde, micri vibratorii precum i spermatozoizi fr micri; Aprecierea s-a
efectuat n funcie numai de micrile rectilinii.
Pentru calculul s-a utilizat formula: R= V/v; n care: R - este rezistena
spermatozoizilor, V - este volumul soluiei de Na Cl 1% folosit n titrare i v
volumul de material seminal (0,02).
Precizm c soluia de NaCl de concentraie 1% a fost preparat
extemporaneu folosind ap bidistilat deionizat. Deionizarea apei bidistilate s-a
fcut cu ajutorul unui aparat Seradest USF ce are la baz un sistem modern de
desalinizare total a apei, printr-un schimb mixt de anioni i cationi.
Observaiile s-au efectuat pe sperma brut, pe sperma diluat proaspt,
dar i pe sperma diluat i conservat n dozele necesare repetrii nsmnarii
artificiale la 24 de ore.
n condiii de izotermie s-a utilizat baia de ap reglat la temperatura de
37C, iar examinarea s-a fcut la un microscop cu fond ntunecat dotat cu platin
nclzitoare, n pictur liber i obiectiv de puterea 10. Examenele au fost
efectuate i la temperatura de 25C.

REZULTATE I DISCUII

Vierii ce au fost luai n studiu aparineau unor rase i linii diferite: 18 din
liniile sintetice: (L 62, L CN, L 19, L 64, L RK) i 5 din rasa Mangalia (graf.1).
Vrstele lor sunt cuprinse ntre 12 i 48 de luni. Programul de utilizare a vierilor
la recoltare a fost ntre 4 i 11 zile (Tabel 1).

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Valorile rezistenei spermatozoizilor obinute la cei 23 de vieri au fost


cuprinse ntre 4500 i 17000, cu o valoare medie de 11564. Valorile minime i
maxime sunt puse pe baza variaiei individuale i a a rasei.
Se observ c la rasa Mangalia valorile rezistenei sunt mult crescute fa
de a celelorlalte rase studiate, iar valoarea cea mai mic s-a nregistrat la vierii din
linia sintetic CN a rasei Landrace.

Tabelul 1
Caracteristicile biotehnolgice ale vierilor luai n studiu

Programul Rezistena S.B.


Rezistena S.D.
Nr. Vrsta de (ml)
Rasa / Linia
Crt. (luni) recoltare Conservat
(zile) 37C 25C Proaspt
24 h
1. Mangalia 18 5 15500 10500 9000 7500
2. Mangalia 13 5 17000 12500 - -
3. Mangalia 31 6 11000 - 9500 -
4. Mangalia 18 6 12000 10000 7500 -
5. Mangalia 15 10 12000 - -
Media 19 6,4 13500 11000 8666 7500
6. Pietrain L62 41 4 13000 - 6000 -
7. Pietrain L62 42 7 10500 - 9500 8500
Media 41,5 5,5 11750 - 7750 8500
8. PIC L62 26 4 13000 - 6000 -
9. PIC L62 22 4 10000 4500 6000 6000
10. PIC L62 18 7 10000 - 8500 8500
11. PIC L62 21 5 11000 - 7000 -
12. PIC L62 25 7 12000 9500 9000 7500
Media 22,4 5,4 11200 7000 7300 7333
13. Pietrain L64 48 4 9000 8500 7500 7000
14. Pietrain L64 21 4 11500 - 7000 -
Media 34,5 4 10250 8500 7250 7000
15. Landrace LCN 13 5 10500 7500 7500 7000
16. Landrace LCN 13 9 10500 - 6500 -
17. Landrace LCN 13 6 8000 6000 5000 -
18. Landrace LCN 12 5 10000 - - -
Media 12,75 6,25 9750 6750 6333 7000
19. Pietrain LRK 15 8 13000 - 11000 9000
20. Pietrain LRK 24 11 14000 - 12500 -
Media 19,5 9,5 13500 - 11750 9000
21. Duroc L19 24 9 10500 8000 7000 6500
22. Duroc L19 24 7 14500 12000 7500 -
23. Duroc L19 24 5 8000 6000 7000 -
Media 24 7 11000 8667 7167 6500

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Duroc L19
Mangalia
13%
21%
Pietrain LRK
9%

Pietrain L62
9%
Landrace
LCN
17%

Pietrain L64
9% PIC L62
22%

Grafic 1 - Distribuia vierilor luai n studiu n funcie de ras/linie

Lucrndu-se n condiii de izotermie (37C), cele mai ridicate valori au fost


obinute pe sperma brut (proaspt recoltat) cu o medie de 11664; valoarea
acesteia a sczut cnd s-a lucrat pe sperm dup diluare, (cu o medie de 8031).
Valorile cele mai sczute au fost obinute pe sperma diluat i pe cea conservat
24 de ore, (media fiind de 7548). Cu ct sperma este examinat ntr-un timp ct
mai scurt dup recoltare cu att valoarea rezistenei este mai ridicat. Prin
conservarea materialului seminal timp de 24 de ore s-a observat o uoar scdere
a valorilor rezistenei spermatozoizilor, dar neconcludent.
Tabelul 2
Media rezistenei spermatozoizilor n funcie de programul de recoltare la
temperatura de 37C

Nr. Programul de Sperm Sperm Sperm


Rasa / Linia
ctr. recoltare (zile) brut diluat conservat 24 h
1. Mangalia 6,4 13500 8666 7500
2. Pietrain L62 5,5 11750 7750 8500
3. PIC L62 5,4 11200 7300 7333
4. Pietrain L64 4 10250 7250 7000
5. Landrace LCN 6,25 9750 6333 7000
6. Pietrain LRK 9,5 13500 11750 9000
7. Duroc L19 7 11000 7167 6500

X Media 6,3 11564 8031 7548

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6500
Duroc L19 7167 11000
9000 11750
Pietrain LRK 13500
7000
Landrace LCN 6333 9750
7000
Pietrain L64 7250 10250
7333
PIC L62 7300 11200
8500
Pietrain L62 7750 11750
75008666
Mangalita 13500
0 2000 4000 6000 8000 10000 12000 14000

Sperma bruta Sperma diluata Sperma conservata 24 h

Grafic 2 - Media rezistenei spermatozoizilor pe categorii de sperm

n condiiile n care temperatura de lucru a soluiei de NaCl 1% a fost de


25C, valorile obinute la determinrile efectuate pe sperma brut au fost n
medie de 8667, constatndu-se o scdere constant la toate rasele/liniile. Diferena
dintre valorile rezistenei determinate la 37C i la 25C poate fi pus pe seama
faptului c temperatura sczut grbete procesul de intoxicare al
spermatozoizilor
Tabelul 3
Influena temperaturii de examinare asupra mediei indicilor de rezisten

La temperatura de La temperatura de
Nr.Crt. Rasa / Linia
37C 25C
1. Mangalita 13500 11000
2. PIC L62 11200 7000
3. Pietrain L64 10250 8500
4. Landrace LCN 9750 6750
5. Duroc L19 11000 8667

X - 11140 8338

Vierii care au avut programul de recoltare la 4 zile au avut valori n medie


de 10250. la un program de 9,5 zile valoarea medie a rezistenei a fost de 13500;
cu ct programul de recoltare a vierilor are mai multe zile de pauz cu att
valoarea rezistenei spermatozoizilor crete.

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16000
14000 13500
12000
11000 11200 11000
10000 10250 9750
8500 8667
8000
7000 6750
6000
4000
2000
0
Mangalita PIC L62 Pietrain Landrace Duroc L19
L64 LCN
La temperatura de 37C
1 2 3 4 5
La temperatura de 25C
Grafic 3 - Influena temperaturii asupra indicilor de rezisten

14000
12000
10000
8000
6000
4000
2000
0
4 zile 5,4 zile 5,5 zile 6,25 6,4 zile 7 zile 9,5 zile
Sperma bruta zile
Sperma diluata
Sperma conservata 24 h

Grafic 4 - Rezistena spermatozoizilor n funcie de media zilelor de recoltare

CONCLUZII

La rasa Mangalia s-au ntlnit cele mai mari valori ale rezistenei
spermatozoizilor la soluiea de NaCl 1%, astfel: valoarea medie a spermei brute a
fost de 13500 la temperatura de 37C i de 11000 la temperatura de 25C; 8666
pe sperma dup diluare; i de 7500 dup conservarea timp de24 de ore.
Diluarea i conservarea materialului seminal scade valoarea rezistenei
spermatozoizilor, dar rmne mult peste limita superioar descris n literatura de
specialitate;

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Optimizarea programului de recoltare a materialului seminal n direcia


creterii lui influeneaz n sens favorabil rezistena spermatozoizilor.

BIBLIOGRAFIE

1. Bogdan A.T., Mantea St., Bogdan Dorina, 1999 Tratat de reproductie si


insamantari artificiale la suine , Ed. Tehnica Agricola, Bucuresti
2. Drugociu D., 2001 Reproducie, nsmnri artificiale i ginecologie veterinar,
Ed.Venus, Iai.
3. Gary C., Althause, 2000 - Optimizing productivity of the AI boar, Department of
veterinary Clinical Medicine University of Illinois Urbana, IL 6802.
4. Manzatu I., Bogdan A.T., 2003 Dictionar explicativ de Biotehnologie , vol. I,
Ed. Global, Bucuresti
5. Nacu G., 2005 Cercetari privind unele posibilitati de optimitare a functiei de
reproductie la suine , Texa de doctorat, USAMV, Iasi;
6. Runceanu L., Nicorici R., 1986 - Patologia reproduciei i clinic
obstetrical , lucrri practice, Iai.
7. Runceanu L., Cotea C., 2001 Reproducie, obstretic i ginecologie
veterinar , Ed. Ion Ionescu de la Brad, Iasi;
8. Runceanu L., Drugociu D., Roca P., Anton C., 2002 - Reproducie,
obstetric i andrologie clinic , Ed. Venus.
9. Kevin J. Rozeboom - Evaluating boar semen quality, Extension Swine specialist
department of animal science North Carolina State University.

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METODE DE DETERMINARE A MOMENTULUI OPTIM


PENTRU MONT SAU NSMNARE LA CEA
OPTIMAL BREEDING TIME AND ARTIFICIAL INSEMINATION MOMENT IN BITCHES,
ESTABILISHED BY DIFFERENT METHODS

Ana- Monica COVACIU*, I. PAPUC**


U.S.A.M.V. Cluj- Napoca

Ovulation appears in the 1-4 estrus day, two days after LH peak
involving the progesterone level increasing before ovulation approximate
value being 5,4 ng/ml. Behavioural changes also appear at this time.
Reproductive tract diseases are caused by many factors concerning genital
tract diseases and also behavioural problems. I tis very important to
determine the ovulation moment to estabilish the optimal mating time or
artificial insemination to increase the number of puppies and decrease the
costs and the stress caused by long distances transport.

Key words: ovulation, estrus, bitch.

INTRODUCERE

Importana reproduciei la specia canin, n ultimii ani, a cunoscut o


cretere spectaculoas, datorit interesului acordat de ctre proprietari acestei
specii, n vederea obinerii unui numr sporit de pui pe un cuib. Acest fapt a
impus specialitilor ca pe lng metodele clasice de depistare a momentului
ovulaiei la cea, cum ar fi: examenul clinic, examenul frotiului vaginal,
examenul ecografic, s fie identificate i utilizate noi metode a cror aplicabilitate
s fie superioar celor clasice. (1, 3) Cteva din noile metode utilizate sunt
reprezentate de : dozarea hormonilor progesteron i LH i msurarea rezistenei
electrice a secreiei vaginale, n scopul supravegherii corecte a estrului la cea.
(4, 6)

MATERIAL I METOD

Lucrarea de fa i propune s determine prin diverse metode, faza


ciclului estral la cea, n vederea unei sincronizri ct mai exacte a momentului
ovulaiei cu monta natural sau nsmnarea artificial (I.A.), precum i
evaluarea performanele metodelor hormonale n comparaie cu celelalte metode
utilizate n mod curent n supravegherea estrului la cea

*
E-mail: covaciumonica@yahoo.com
**
E-mail: i_papuc@yahoo.com

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n scopul efecturii acestui experiment s-au utilizat un numr de 4 cele


adulte din rasele: Rotweiller (1 ), Labrador (1 ), Amstaff (1 ), Ciobnesc
german (1 ), cu vrste cuprinse ntre 3 i 5 ani, care au fost aduse la consultaie
n clinica de reproducie a facultii. Experimentele s-au efectuat n perioada
Aprilie 2003 Mai 2005.
Ca metode de lucru au fost utilizate:
- Observaii clinice i comportamentale asupra femelelor n clduri
- Efectuarea de frotiuri vaginale n fiecare etap a ciclului sexual
- Determinarea concentraiei hormonilor: LH i progesteron.

REZULTATE I DISCUII

Pe baza examenului clinic s-au urmrit modificrile de la nivelul aparatului


genital i s-a ntocmit etograma comportamentului sexual n perioada de clduri.
La debutul ciclului, n primele 4-7 zile, femela devine nelinitit, agitat, sociabil
fa de masculi, apoi vulva ncepe s se tumefieze progresiv. ncepnd cu ziua a
opta, femela prezint o congestie puternic a mucoasei genitale aparente, nsoit
de un mucus clar, filant, uneori sangvinolent.
Etograma comportamentului sexual la cea i modificrile organelor
genitale externe:

Fig. 1, 2 - Modificrile organelor genitale externe

Efectuarea de amprente vaginale i examenul frotiului vaginal:

Fig. 3. Frotiu vaginal, debut Fig. 5. Optim pentru mont


Fig. 4. Frotiu vaginal estru.
de estru sau .A.

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Realizarea frotiului citovaginal include urmtoarele etape:


- Prelevarea
- Etalarea
- Colorarea
- Metoda de colorare cu albastru de metilen
- Metoda de colorare cu albastru policrom
La debutul estrului, frotiul prezint numeroase celule poliedrice cu nuclei
picnotici sau fr nucleu, dispuse n grmezi (celule Scholen). Nu se gsesc celule
parabazale sau intermediare. E posibil ca n frotiu s fie surprinse i cteva
hematii.
n estru frotiul prezint numeroase celule poliedrice cu nuclei picnotici sau
fr nuclei, dispuse n grmezi, lipsesc celulele parabazale i intermediare mici.
Dozri hormonale
Momentul optim al ovulaiei poate fi determinat i prin dozarea hormonilor,
mai ales a LH-ului (care indic exact stadiul n care se afl ovula) i a
progesteronului
Concentraia celor doi hormoni este sczut n timpul proestrului. FSH ul
i LH ul ating vrful concentraiei la nceputul estrului, inducnd ovulaia
Determinrile hormonale au fost efectuate la Laboratorul SYNEVO din
cadrul Institutului Oncologic Cluj-Napoca
Probele au fost prelevate n zilele 1, 3, 5, 7, 9, 11, 13 ale perioadei de
clduri
Toate dozrile s-au efectuat dup o supraveghere a ciclului estral prin
metoda frotiului citovaginal
Tabelul 1
Cazuri luate n studiu

Nr.crt Nume femel Ras Vrst Debutul cldurilor


1 Kimba Amstaff 4 ani 19.03.2005
2 Alfa Ciobnesc german 3 ani 10.02.2005
3 Rita Labrador 4 ani 22.04.2005
4 Linda Rottwailer 5 ani 04.02.2005

Tabelul 2
Valori obinute

Ziua Valorile progesteronului


1 0,19 ng/ml
3 0,308 ng/ml
5 1,96 ng/ml
7 2,34 ng/ml
9 3,08 ng/ml
11 7,86 ng/ml
13 7,92 ng/ml

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Fig. 6 - Buletin de analiz, caea 3 ani, ziua 11 a ciclului

CONCLUZII

Stabilirea momentului optim de ovulaie la celele de mare valoare


biologic, att din canise ct i de la cresctorii individuali, constituie o prioritate
pentru a evita pierderile de gestaie.
Depistarea momentului optim pentru mont la cea, este recomandat a se
face prin coroborarea datelor obinute la examenul comportamentului sexual, pe
baza frotiurilor vaginale efectuate seriat (din prima zi pn n ultima zi a
cldurilor) i a determinrilor hormonale.
Utilizarea dozrilor hormonale n depistarea momentului ovulaiei la cea
sunt superioare celorlalte metode utilizate la aceast specie, fapt ce determin
creterea numrului de pui/mont
Metodele permit aprecierea concentraiei serice de LH i progesteron, n
consecin ofer date importante n ceea ce privete dinamica funciei ovariene,
alturi de examenele clinice clasic.
Pe baza frotiurilor citovaginale i a determinrii progesteronului i LH-ului
se recomand efectuarea montei naturale sau I.A. n zilele 9-13 ale ciclului estral .

BIBLIOGRAFIE

1. Botrel, S., C., Cotea, M., Gaboreanu, 1982 Histologie i embriologie, Editura
Didactic i Pedagogic Bucureti.

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2. Boichard, G., Youngquist, R.S., et al, 1991 - Seasonality and variability of the
interestrous interval in the bitch Therio 36: 41-50.
3. Cociu, M., 1999 - Etologia i comportamentul animal, Editura.All Bucureti.
4. Feldman, Nelson, 1996 - Ch 22 Infertility, associated breeding disorders and
disorders of sexual development, In: Canine and Feline Endocrinology and
Reproduction, W.B. Saunders, Philadelphia P.A., (619- 648).
5. Groza I., Muntean, M., 2002 - Elemente de fiziologia reproduciei la animale, Editura
AcademicPres Cluj- Napoca.
6. Kolb, E., 1981 - Despre viaa i comportamentul animalelor domestice, Editura
Ceres Bucureti.

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OBSERVAII PRIVIND INFLUENA SALPINGITELOR


ASUPRA UNOR PARAMETRI DE REPRODUCIE LA VACI
OBSERVATIONS REGARDING SALPINGITIS INFLUENCE ABOUT CERTAIN
REPRODUCTION PARAMETERS OF COWS

I. DOROBANU, C. GALAN

The investigations have been performed on 215 cows regarding the


salpingitis weight in reproduction pathology and their influence on certain
synthetic reproduction parameters. A number of 18 cases were diagnosed
with tubar inflamation (8,4%).
The regestred Service Period had a value between 170 and 500
days. Accordingly increased values of Calving Interval Indicator (until 785
days) and Pregnacy Index were rgistred.

Key words: reproduction, salpingitis, cow

MATERIAL I METOD DE LUCRU

Observaiile efectuate pe 215 femele crescute n sistem industrial au


urmrit: anamneza, examenul ginecologic extern i examenul clinic intern
colposcopic i transrectal, n vederea stabilirii ponedrii salpingitelor.
Examenul clinic intern s-a refrit la:
- inspecia vulvei i regiunii perivulvare
- eliminarea i caracterul secreiilor
- permanena sau intermitena scurgerilor uterine.
Examenul clinic intern, manual sau colposcopic.
La vac, examinarea segmentelor componente ale tractusului genital s-a
efectuat ntr-o anumit ordine : gtul uterin, corpul uterin, coarnele uterine,
oviductele i ovarele. n aceai manier, s-a acordat atenie ligementelor largi,
mai ales n cazurile n care se constat deformri ale zonei perisalpingiene.
n mod normal, la femelele sntoase, gtul uterin este destul de lung i
plasat pe planeul cavitii pelvine; este uor deplasabil i consistent. Pierderea
n elasticitate i consisten poate fi identificat n caz de gestaie, pe cnd
creterea consistenei denot o cervicit cronic.
Coarnele uterine s-au examinat sub aspect topografic, al consistenei,
sensibilitii i a tonusului. n cazul endometritelor s-a observat lipsa reaciei la
palpaie i o uoar indurare.
Oviductele, la cazurile examinate de noi s-au prezentat mult ngroate,
pn la dimensiunea de aproximativ 0,5 cm, urmnd traectul sinuos caracteristic
(mai ales n formele acute). Alteori au avut aspectul unor cordoane de consisten
dens, cu unele aderene (n salpingitele cronice).
Se apreciaz:
- caracterele mucoasei vaginale

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- forma pliurilor, care n general sunt terse, dar pot deveni evidente n
inflamaiile cronice uterine
Dat fiind c cervicitele sunt adeseori cauze de infecunditate s-a procedat la
explorarea segmentului vaginal al gtului uterin. n acest sens s-a acordat o
atenie deosebit la volumul gtului uterin, topografia, forma, abundena i
caracterul secreiilor.

REZULTATE I DISCUII

Onservaiile noastre privind incidena ginecopatiilor au scos n eviden


mai multe entiti patologice localizate att la nivelul gonadelor ct i a tractusului
genital.
Tabelul 1
Incidena ginecopatiilor

Femele examinate Diagnostic prin tehnica


Specificare
Nr. % insuflrii de aer
Endometrite 65 30,1
Distocii 20 9,4
CL persistent 20 9,4
Chiti ovarieni 40 18,5
Salpingite unilaterale 31,43
Salpingite 18 8,4
Salpingite bilaterale 17,14
Cervicite 17 8,0
Retenia anexelor fetale 35 16,2
Total 215

Examinarea a 215 femele a scos n eviden faptul c cel mai ridicat


procent a fost ntlnit n cazul endometritelor (65 cazuri, reprezentnd 30,1%). n
ordinea frecvenei urmeaz boala chistic ovarian (40 cazuri, reprezentnd
18,5%), urmate de retenia anexelor fetale (35 cazuri, reprezentnd 16,2%).
Afeciunile ovariene au reprezentat un procent egal cu distociile 9,4%.
Salpingitele, diagnosticate numai prin examen clinic intern transrectal, au
reprezentat 8,4% (18 cazuri). Remarcm faptul c diagnosticul clinic nu
precizeaz n totalitate existena unor salpingite i mai ales a obstruciilor
salpingiene considernd c este necesar aplicarea i a altor metode n vederea
precizrii diagnosticului.
n acest sens, datele comunicate de ctre Khasatya 1999 arat c
diagnosticul prin tehnica insuflrii de aer n vederea precizrii permeabilitii
oviductelor pune n eviden un procent mai mare dect examenul clinic de rutin.
Astfel, autorul evideniaz prin aceast tehnic salpingite unilaterale ntr-o
proporie de 31,43% iar salpingitele bilaterale ntr-o proporie de 17,14%.
Considerm n acest caz c diagnosticul numai prin examen clinic nu
evideniaz n totalitate cazurile de salpingit sau obstrucie tubar.

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Observaiile noastre au urmrit i evoluia ciclurilor sexuale n diverse


ginecopatii. Au fost investigate mai multe elemente:
- intervalul de repetare a estrului (zile)
- SP service period
- CI calving interval
- IG indice de gestaie
Tabelul 2
Evoluia ciclurilor sexuale n ginecopatii

Interval de repetare a
Specificare SP CI IG
estrului (zile)
Endometrite acute 70 204,3 489 6
Endometrite cronice 70-160 249 534 14
CI persistent - 200 485 -
Chiti foliculari 9-18 90-120 375-405 12
Salpingite acute 21-28 170 445 16
Salpingite cronice Neregulat ntre 40-300 500 785 8
Retenia nvelitorilor fetale 48-60 68 353 4

Legend: S.P. = Service Period ; C.I. = Calving Interval ; I.G. = Indice Gestaie

Apreciate pe baza parametrilor de reproducie influena diverselor


ginecopatii este diferit n funcie de forma clinic. Astfel, observaiile noastre
scot n eviden urmtoarele:
- intervalul parturiie fecundaie este foarte variabil n cazul
salpingitelor, n unele cazuri (cronice) depind 500 zile.
- intervalul dintre dou parturiii este cu mult peste asigurarea unei
rentabiliti a exploatrii acestor female: cel mai mic (salpingitele
acute 445 zile).
- indicele de gestaie ajunge la valori foarte mari mai ales n formele
clinice acute (16).
- intervalul de repetare a estrului este foarte variabil, n unele cazuri,
funcionabilitatea ovarian fiind cel puin parial pstrat.

CONCLUZII

1. Observaiile noastre effectuate pe 215 femele au artat o inciden de


8,4% salpingite, diagnosticate prin examen clinic. Procentul observat de noi prin
metodele clinice este mult mai mic dect diagnosticul prin metode de verificare a
permeabilitii tubare asociat cu examenul clinic.
2. Analiza unor parametric de reproducie privind intervalul de repetare a
estrului, SP, CI, i IG arat unele diferene ntre formele clinice diagnosticate.
3. Intervalul de repetare a estrului n cazul salpingitelor a fost cuprins ntre
21-28 zile (formele acute) i 40-300 zile (n formele cornice).

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4. SP a fost cuprins, n funcie de forma de evoluie, ntre 170-500 zile cu


maximum n formele cronice.
5. CI a realizat un interval de 785 de zile n formele de salpingit cronic,
nregistrndu-se tot odat un IG cuprins ntre 8-16.
6. Cervicitele s-au caracterizat printr-un SP de 118 zile, un CI de 403 zile i
IG de 11.

BIBLIOGRAFIE

1. Ardelean, V., 1998 Reproducia animal. Ed. Agroprint. Timioara.


2. Bogdan, A.T., 1984 Fertilitatea, natalitatea i prolificitatea n zootehnie. Ed. Dacia,
Cluj.
3. Boitor, I., 1981 Sterilitatea la femelele animalelor de ferm. Ed. Dacia, Cluj-
Napoca.
4. Cernescu, N., 2004 Ginecologia veterinar. Ed. Cecma, Timioara.
5. Drugociu, D., 2004 Bolile obstetrical ginecologice la animale.
6. Drugociu., Gh., 1977 Patologia reproduciei i clinic obstetrical. Ed. Didactic i
Pedagogic Bucureti.
7. Gluhovschi, N., 1970 Ginecologie veterinar. Ed. Ceres, Bucureti.
8. Hafez, E.S.E., 1987 Reproduction in farm animals.
9. Runceanu, L., Cotea, C., 2001 Reproducie, obstetric i ginecologie veterinar.
Ed. Ion Ionescu de la Brad Iai.
10. Seiciu, Fl., i col., 1987 Reproducia normal i patologic la animalele domestice.
Ed. Ceres, Bucureti.
11. Thibault, Ch., 1991 La reproduction chez les mamiferes et lhome. Ed. Marketing
Paris.

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THE DIAGNOSTIC IMPORTANCE OF OVARIAN AND


ADENOHYPOHYSIS DOSAGE IN DAIRY COWS OVARIAN
FUNCTIONAL DISORDERS
D. DRUGOCIU, L. RUNCEANU, P. ROCA
U.S.A.M.V. Iai

The results of the hormonal dosages in females having ovarian


hypotrophy of I degree, emphasized an ovarian hypofunction by
hyposecretion adenohypophyseal, having a F.S.H., L.H. and progesterone
secretory insufficiency and an increase of the prolactin level.
The same situation was registered in the cows presenting ovarian
hypotrophy of II degree.
In the females diagnosticated with persistant luteal corps the value
of F.S.H./L.H. ratio is almost fair. In the females presenting the I.A.
repetition syndrome, the F.S.H./L.H. ratio was of 1.29/l for F.S.H. so, we
can say that the main factor of repeated I.A. is an adenohypophyseal
hyposecretion of L.H.

Key words: cows, reproduction, hormonal, dosage.

Lately there were many concerns in elucidating the possibilities and limits
of using hormonal therapy in animal reproduction proces (1, 3, 7).
Some of the practitiones specialists are sceptical in using hormonal therapy,
because of the female individual reactions after the therapy.
These reactions may appear due to the fact that there is no differential
therapy, depending on the degree of the functional disorder (2, 6).
In order that these negative aspects of hormonotherapy not to appear it is
indicated thet the clinical diagnosis be corroborated with the laboratory one (the
hormonal).
Once the titre of the gonadotropic hormones and of the sexual hormones is
established, depending on their level the hormonal therapy can be applied (4, 5,
8).

MATERIAL AND METHODS

In order to perform the hormonal dosage, blood samples have been taken,
depending on the diagnosis, after the gynaecology investigation and the clinical
diagnosis are made.
In accordance with the clinical diagnosis, the females have been grouped
in desease categories: dairy cows with ovarian hypotrophy (stage I and II),
females with repeated artificial insemination and females with persistent corpus
luteum.

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The blood sample has been proccesed after the usual technique in order to
obtain the blood serum.
To establish the hormonal diagnosis it was followed the progesterone
dosage, LHs (luteinizing hormone), FSHs (follicle stimulating hormone) and the
seric prolactins.
The seric progesterone tests determination was performed with the ELISA
test, using streptanidina technology. The dosage of the LH, FSH and seric
prolactin levels was performed through the ELISA sandwich test in one step, with
amplified signal.

RESULTS AND DISCUSSIONS

The hormonal dosage at the females diagnosticated with stage I ovarian


hipotrophy, shows an ovarian hypofunctions through adenohypophysis
hyposecretion. This is caused by a secretory insufficiency of the adenohypophysis
hormones, FSHs and LHs.
The titre of the gonadotropic hormones, produces an inhibation of the
gonadotropin releasing factors (gonadotropin LH/FSH Releasing Hormons).
Due to this fact the low level of the LH at dairy cows with stage I ovarian
hypotrophy was 2,12 m UI/ml, and FSHs was 4,02 m UI/ml at the inferior limit
of the basic level. At the same females it was established a low medium level of
progesterone and a high level of prolactin.
6,24 mg/ml. Because of the high titre of prolactin and a
hypoprogesteronemie (an average of 1,07 mg/ml) it causes the block of the sexual
cicle, with the instalation of the anestrus.
A similar situation was found at a group of females clinical diagnosticated
with stage II ovarian hypotrophy. So at these females the medium level of seric
progesterone is low (0,23 mg/ml), the FSHs at the basic minimum level (1,08 m
UI/ml) adn the prolactins of 2,7 mg/ml. Clinical, these females diagnosticated
with stage II ovarian hypotrophy, showed an prolonged state of anestrous.
Comparing these results with the ones from the witness lot it was noticed
that at the females with stage II hypotrophy, the seric concentration of the
progesterone (0,23 mg/ml), prolactins and titres (1,08 m UI/ml) was significant
smaller, producing this way severe modifications alterations to the genital
apparatus.
In case thet corpus luteum persists the decisive factor is the value of the
medium raport FSH/LH (1,28/1m UI/ml) that is almost equal to the high medium
level of the prolactin (3,59 mg/ml). To all these factors it adds a secretory
insufficiency of PGF2 (prostaglandin 2 alpha), that causes the installation of an
prolonged state of anestrous. That is why it is recommended that at the females
diagnosticated with an persistent corpus luteum to be administrated a dose of
PGF2 .

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At the females, clinical diagnosticated with artificial insemination


repeated sindrom, was noticed that the statistic value media of the FSH is 3,65,
and LHs is 2,81. Following the statistic calculation, the value of the FSH/LH
report is 1,29/1 in favour of the FSH. So it can be estimated thet the primary
cause of the repeated A.I. is an adenohypophysis hyposecretion of LH. Because of
this restoring the physiological report between FSH and LH (1/3, so thet the
ovulation can take place) it has to be administrated, to the females, products with
LH effect, a few hours before the A.I.

CONCLUSIONS

1. At the dairy cows, clinical diagnosticated with stage I and II ovarian


hypotrophy, was established an ovarian hypofunction with adenohypophisis
hyposecretion.
2. To resume the genital activities it is recommended thet to the dairy cows
with ovarian hypotrophy be administrated products with FSH or PMSG in dose of
1000 UI/animal.
3. In case of the females with A.I. repeated sindrom because of the
hormonal dosage, it was established an adenohypophysis hyposecretion of LH.
4. To restore the normal titre of LH and start the ovulation it is
recommended thet those females be administrated products with LH effect (HCG
1500 U.I.; Gn-RH 2,5 ml)

BIBLIOGRAPHY

1. Aguer, D., 1981 Les progestagenes dans la matrise des cycles sexuels chez les
bovins. Rec. Med. Vet., 157(1) : 53-60.
2. Broiu, A., Seiciu, Fl., 2002 Ginecologie i andrologie veterinar. Ed. Fundaiei
Romnia de mine.
3. Brueuel, K.F., Lewis, P.E., 1990 Endocrine profiles and follicular development in
early weaned postpartum cows. Journal of Reprod. and Fertility, vol. 97, no. 1.
4. Galvin, J., Ryan, D.P., Boland, M.P., Roche, J.F., Yaakub, H., Snyders, S.,
OFarrel, K.Y., 1995 Ovarian follicular development following progestiv and
gonadotropin treatments at varying stages of the follicular wave and oestrus
cycle. J. Reprod. Fertil. Abstract, 15, 13.
5. Gong, J.K., Campbell, B.K., Bramley, T.A., Peters, A.R., Webb, R., 1955
Evolution of the requirements for F.S.H. and L.H. during ovarian follicle growth
and development in cattle. J. Reprod. Fertil. Abstract, 15,7.
6. Nelis, P.C., 1995 Compendium of Animal Reproduction Publisher Intervet
International B.V. Second edition.
7. Paraipan, V., 1982 Hormonoterapia n reproducia animal. Ed. Ceres, Bucureti.
8. Parez, V., 1994 Matrise de la reproduction dans lespece bovine. Laction
veterinaire LHV. Supplement An. Nr. 1277.

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OBSERVAII ASUPRA FRECVENEI ENDOMETRITELOR


I CONSECINELE ACESTORA N PATOLOGIA
REPRODUCIEI LA VAC
ENDOMETRITIS FREQUENCE OBSERVATIONS AND THEIR RESULTS ON
REPRODUCTION PATHOLOGY OF COWS

C. GALAN, I. DOROBANU

The acute endometritis diagnosed especially in the puerperal period


were represented by two clinically dominant forms : catarrhal and purulent.
The observations were about the diseases dynamic in three years
period, the ratio of chronic and acute forms and the evolution of the chronic
endometritis as a continuation of undiagnosed acute inflammations or
inadequate treated.
It comes out that the acute forms of purulent endometritis represents
8,03% cases and catarrhal endometritis 5,12%. The recoveries of acute
forms have been catharrhal forms and 76,3% in purulent forms.

Cuvinte cheie: Reproducie, vac, endometrit.

Infertilitatea femelelelor nu reprezint o boal propriu-zis, ci un sindrom


corespunztor diferitelor tulburri ale fucniei de reproducie. (Drugociu, Gh.,
1977, Gluhovschi, N., 1971, Seiciu, Fl., 1987, Runceanu, L., Cotea, C., 2001,
Cernescu, H., 2004, Broiu, A., 2004, Huba, I., 1997).
Infertilitatea la vac poate fi o consecin a numeroase cauze (plgi,
anomalii, inflamaii, etc), ncepnd cu perioada puerperal.
Endometritele sunt inflamaii ale mucoasei uterine, ce pot apare primar, sau
ca urmare a infeciei unui segment genital nvecinat, de cele mai multe ori pe cale
ascendent prin contiguitate.
Infeciile uterine la bovine, grupeaz toate strile patologice, produse de
agenii patogeni n perioada puerperal, respectiv, pn la 21 zile (Seiciu, Fl.,
1987). Infeciile ce depesc aceast perioad sunt (Martius, H., 1986) considerate
a fi trecut n forma cronic de evoluie.
n funcie de intensitatea reaciei inflamatorii, apare o gam variat de
exprimri clinice. Endometrita cataral acut este diagnosticat de cele mai multe
ori n perioada puerperal. Boala apare la 8-12 zile de la parturiie, n funcie de
ritmul involuiei uterine, puterea de aprare local i gradul de patogenitate al
germenilor. Evoluia endometritei catarale acute poate urma calea vindecrii n
decurs de 8-10 zile, timp n care secreia scade cantitativ, se clarific treptat, iar
involuia uterin progreseaz. n multe cazuri ns endometrita acut se
cronicizeaz, iar forma cronic evolueaz extrem de insidios sau se transform n
endometrita purulent.

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Endometrita purulent acut se caracterizeaz prin inflamaia endometrului


i acumularea de mucopuroi n cavitatea uterin. n aceste cazuri, starea general
se altereaz, apare febra, diminuarea apetitului i a secreiei lactate. Pe ovare se
palpeaz corpul luteal persistent (Muntean, C., 2005)

MATERIAL I METODE DE LUCRU

Observaiile noastre au fost efectuate n perioada 2003-2005 lundu-se n


eviden un numr de 578 femele, apreciindu-se frecvena endometritelor acute
(cataral i purulent), forma morfoclinic a acestora precum i posibilitatea
cronicizrii inflamaiei.
Diagnosticul a fost stabilit prin examen ginecologic i completarea fiei
ginecologice : anamneza, examenul clinic extern (modificri ale organelor
genitale extern i caracterul secreiilor), examenul clinic intern (colposcopic i
transrectal). Prin examen colposcopic s-au evideniat modificrile mucoasei
vaginale i cervicale, culoarea, forma i aspectul pliurilor, natura secreiilor.
Examenul transrectal a urmrit evidenierea componentelor tractusului
genital (forma, mrimea, consistena, topografia, precum i starea funcional a
ovarelor).
Tabelul 1
Apariia endometritelor cronice ca o continuare a endometritelor acute

Endometrita acut Endometrita cronic Endometrita


acut vindecat
Anul Cataral Purulent Cataral Purulent dup tratament
Nr. %1 Nr. %1 Nr. %2 Nr. %2 Nr. %2
2003 31 5,38 5 16,1 1 3,2 25 83,3
59 10,26 8 13,5 6 10,1 45 76,4
2004 40 5,97 7 17,5 2 5 31 77,5
60 8,95 8 13,3 3 5 49 81,7
2005 20 4,08 4 20 1 5 15 75
24 4,89 5 20,8 2 8,3 17 70,9
Media 27 5,12 5 17,8 1 4,4 23 78,6
pe cei 3
ani 47 8,03 7 15,4 3 7,8 37 76,3

REZULTATE I DISCUII

Din analiza datelor, s-a nregistrat un numr de 75,3 cazuri (pe cei trei ani):
114 n anul 2003, 123 n anul 2004 i 60 n anul 2005.
Numrul crescut de cazuri n anii 2003-2004 se datoreaz n special
alimentaiei necorespunztoare din punct de vedere cantitativ i calitativ.
Din datele tab. 1 constatm c numrul mediu de cazuri cu endometrit
cataral acut a fost de 27, reprezentnd 5,12%. Se observ un numr relativ

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constant al acestei forme n anii 2003 i 2004: 5,38% i respectiv 5,97%, fiind
ceva mai sczut n anul 2005.
Forma acut purulent nregistreaz o medie de 47,6 cazuri. Procentual,
fa de valorile medii (8,03%), se constat variaii ntre 10,26 n anul 2003, 8,95
n 2004 i 4,89 n anul 2005.
Urmrind evoluia endometritelor acute care se pot croniciza, constatm, n
dinamica anilor 2003-2005 c n unele cazuri formele acute catarale sau
purulente, se cronicizeaz.
Astfel, n anul 2003 au fost nregistrate un numr de 31 cazuri cu
endometrit acut cataral , reprezentnd 5,38% din totalul femelelor examinate.
Dintre acestea 83,3% s-au vindecat, 16,1% au trecut n forma cataral cronic, iar
3,2% n forma purulent cronic.
n anul 2004 au fost diagnosticate 40 cazuri (5,97%) cu endometrit
cataral acut ; dintre acestea 31 cazuri (76,4%) s-au vindecat, 7 cazuri (17,5%)
au trecut n forma cataral cronic, iar 2 cazuri (5,0%) s-au transformat n
endometrit cronic purulent.
Rezultate asemntoare sunt nregistrate i n anul 2005.
Observaiile privind modificrile formei clinice de endometrit acut
purulent arat c n anul 2003 au fost diagnosticate 59 cazuri (10,26%), dintre
care 45 s-au vindecat, 8 (13,5%) au trecut n forma cataral cronic, iar 6 cazuri
(10,1%) s-au cronicizat.
Date asemntoare se nregistreaz i n anul 2004: 8,95% femele
diagnosticate, 81,7% vindecri iar 5,0% s-au cronicizat. n anul 2005, dinamica
evoluiei clinice este apropiat de anii anteriori.

CONCLUZII

1. Forma morfoclinic de endometrit purulent acut a fost cea mai


frecvent diagnosticat (8,03%), urmat de endometrita cataral acut (5,12%).
2. Dinamica evoluiei acestor afeciuni n anii 2003-2005 arat o asemnare
n procentul de vindecri sau recidive.
3. Trecerea formei clinice din acut n cronic (cataral sau purulent)
arat, importana deosebit a precizrii diagnosticului i instituirea unui tratament
judicios, ntruct n medie 21,8% din formele catarale acute i 23,2% din formele
purulente acute, se transform n forme cronice.
4. Tratamentul femelelor cu endometrit dup aceiai formal terapeutic,
fr a ine seama de variabilitatea florei patogene i individualizarea cazurilor,
complic unele forme acute fr o mare gravitate, n forme cronice, uneori
ireversibile.

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BIBLIOGRAFIE

1. Broiu, A., 2004 Tratat de reproducie la animale. Ed. ALL Bucureti


2. Cernescu, H., 2004 Ginecologie veterinar. Ed. CECMA Timioara.
3. Drugociu, Gh., 1977 Patologia reproduciei i clinic obstetrical. Ed. Didactic i
Pedagogic Bucureti.
4. Gluhovschi, N., 1971 Reproducia animalelor domestice. Ed. Ceres, Bucureti.
5. Huba, I., 1997 Corelation between milk yield and fertility of dairy cows. Zivocisna
282 Slovakia.
6. Mialot, I.P., 1989 Pathologie de la Reproduction chez les carnivores domestiques.
Ed. Du Point Veterinaire Paris.
7. Runceanu, L., Cotea, C., 2001 Reproducie, obstetric i ginecologie veterinar.
Ed. Ion Ionescu de la Brad Iai.
8. Runceanu, L., 1995 Patologia reproduciei i clinic obstetrical. I.A. Iai.
9. Seiciu, Fl., 1987 Reproducia normal i patologic la animalele domestice. Ed.
Ceres, Bucureti.
10. Vaissaire, I.P., 1977 Sexualite et reproduction des mammiferes domestiques et de
laboratoire. Ed. Maloine, Paris.

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STUDIU PRIVIND INCIDENA MAMITELOR CLINICE


LA VACILE CU LAPTE DE RAS BNR
STUDY REGARDING THE INCIDENCE OF CLINICAL MASTITIS
AT DAIRY COWS BNR BREED

N. HROMEI
A.N.S.V.S.A. - PIF Sculeni
Mariana SOFRONIE, M. PNTEA,
Elena RUGINOSU, C. BDELI,
Anca LUPU
S.C.D.B. DANCU Iasi

The study made in a farm of BNR breed milk cows, belonging to


SCDB Dancu, during the period 2001-2005, on a number of 2326 cows. Out
of the total of 588 sacrificed cows, it was observed that a percentage of 8 %
was represented by the the chronic (clinical) mastitis.
The analysis made was related to the percentage of representation of
clinical mastitis depending on the season, stabulation season and lactation
range.
Thus the highest incidence of clinical (chronic) mamitis takes place
in spring (38%) and the lowest in winter (17%). The highest values (23,7%)
are record for the 4th and 5th lactation and the lowest (10,5%) for the 2th
lactation.
The preventing sanitary-veterinary actions, precocious diagnosing
and treatment administering according to optimum timing lead to
diminishing loses in cow farms.

Words key: dairy cows, chronic (clinical) mastitis, season, loses.

Pstrarea nemodificat a compoziiei chimice a laptelui i mai ales


asigurarea salubritii lui, constituie o condiie esenial pentru biosecuritatea
alimentar, cu privire la sntatea uman.
Laptele este un mediu favorabil dezvoltrii unei game variate de
microorganisme. Unele dintre acestea, care pot fi patogene i contagioase se
elimin la nivelul glandei mamare, mai ales cnd aceasta prezint leziuni i
exercit o aciune nociv asupra consumatorilor. n cazul mamitelor clinice s-a
constatat c, frecvent vacile rmn purttoare de germeni , cu un grad foarte mare
de risc de diseminare a bolii n efectivul de vaci din ferm, reprezentnd un
pericol deosebit de grav pentru sntatea animalelor, salubritatea produselor
lactate i sntatea consumatorilor umani (1, 2, 3).
Intensivizarea produciei de lapte presupune o cretere proporional a
capacitii individuale de lactogenez. Pentru a se asigura eficiena economic a

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fermei, cresctorii de vaci, n colaborare cu serviciul sanitar veterinar trebuie s


acioneze n direcia creterii produciei de lapte, ridicarea calitii laptelui i s
supravegheze starea de sntate a glandei mamare a vacilor cu lapte printr-un
program adecvat de prevenire i tratament (4, 5).
Numeroase studii de specialitate evideniaz faptul c, mamitele la vacile n
lactaie produc pierderi economice deosebit de pgubitoare, estimate la 100-140
$/vac, prin scderea produciei de lapte, eliminarea laptelui vacilor tratate cu
antibiotice, scderea calitii laptelui, costul medicamentelor, costul forei de
munc suplimentare, serviciile veterinare i reformarea timpurie a vacilor (1, 2, 3,
5, 9).
Diferite studii de specialitate au indicat o frecven variabil a mamitelor
clinice la vacile cu lapte, de la o ferm la alta, n relaie cu aria geoclimatic,
practicele manageriale i cu ali muli factori tehnologici, terapeutici, de
zooigien, fapt ce impune o monitorizare permanent la nivelul fiecrei ferme de
vaci pentru a se lua msurile cele mai potrivite de reducere a acestor pierderi att
de costisitoare din punct de vedere economic.
Scopul lucrrii este de a efectua o analiz mai complex la nivelul unei
ferme de cretere i exploatare a taurinelor de rasa BNR privind dinamica
manifestrii mamitelor clinice la vacile n lactaie, n relaie cu unii factori de
variaie.

MATERIAL I METOD

Studiul a fost efectuat n perioada 2001-2005 n ferma de cretere i


exploatare a vacilor de lapte, de rasa BNR a SCDB Dancu Iasi, pe un efectiv rulat
de 2326 capete.
Din analiza efectuat a rezultat c animalele au beneficiat de condiii n
general bune de furajare i exploatare, starea de ntreinere fiind apreciat ca
bun i foarte bun.
Nucleul existent n ferm este reprezentat de vaci de nalt valoare
genetic, care nregistreaz producii de 5000-8000 l / vac/ lactaie normal i o
producie medie de lapte n anul 2005 de 6700 l. / vac / lactaie.
Activitatea de prevenire i control a mamitelor a fost bine organizat,
primvara i toamna efectundu-se controlul strii de sntate a glandei mamare
i a calitii laptelui printr-un test rapid de grajd cu reactiv R. Mastitest (produs de
Institutul Pasteur Bucureti). Cu toate aceste msuri s-au nregistrat unele
deficiene n sistemul de exploatare, ct i n cel de colectare a laptelui, care
direct sau indirect au condus la apariia i manifestarea unor afeciuni ale glandei
mamare cu implicare n cantitatea de lapte muls, calitatea laptelui, genernd
unele pierderi la nivel economic.
Pentru studiul iniiat s-a efectuat o anchet epidemiologic, lundu-se n
considerare datele oficiale din unitate. Atfel, pentru perioada 2001-2005 s-au
ierarhizat ieirile din efectiv a vacilor sacrificate pe diferite cauze i s-au analizat

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cele datorate afeciunilor glandei mamare stabilind dinamica incidenei mamitelor


clinice n funcie de variate cauze, cum sunt: anotimpul, sezonul, rangul lactaiei.

REZULTATE I DISCUII

Rezultatele obinute sunt prezentate n tabelele i graficele de mai jos.


Analiznd datele din tabelul 1 se constat c la efectivul rulat de 2326 vaci
n perioada 2001-2005, un numr de 588 capete, reprezentnd 25,3% au fost
sacrificate din diverse cauze. Din totalul sacrificrilor un numr de 47 capete
(8,0%) au avut drept cauze mamitele clinice, entiti patologice care nu au rspuns
favorabil la tratamentele aplicate sau care au recidivat la anumite intervale de
timp (tab.1, fig. 1).
Tabelul 1
Date privind sacrificrile de necesitate la vacile de lapte din diverse cauze
(2001-2005)

Cauza sacrificarii
Total Afectiuni Afectiuni
Efectiv animale
Anul matca sacrificate ale podale si Mamite Afectiuni
Diverse
mediu aparatului ale clinice medicale
genital membrelor
n % n % n % n % n % n %
2001 522 77 14,7 32 41,6 23 29,9 6 7,8 12 15,6 4 5,2
2002 446 84 18,8 43 51,1 18 21,5 7 8,3 11 13,1 5 6,0
2003 451 171 37,9 52 45,6 78 45,6 11 6,4 21 12,3 9 5,3
2004 447 139 31,1 51 36,7 43 31,0 15 10,8 25 18,0 5 3,6
2005 460 117 25,4 42 35,9 47 40,2 8 6,8 20 17,1 0 0
Total 2326 588 25,3 220 37,4 209 35,5 47 8,0 89 15,1 23 3,9

Fig. 1 Dinamica sacrificrilor de necesitate ntr-o ferm de vaci cu lapte (2001-2005)

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Fig. 2. Valorile medii ale sacrificrilor de necesitate cauzate de mamitele clinice la vaci
(2001-2005).

n figura 2 sunt prezentate valorile medii ale sacrificrilor de necesitate


datorate mamitelor cronice n raport cu alte cauze care au determinat ieirile din
efectiv a vacilor. Din totalul iesirilor din efectiv, mamitele clinice ocup locul
patru, dup afeciunile aparatului genital (37,4%), afeciunile membrelor (35,5%)
i afeciunile medicale (15,1%).
Analiznd incidena mamitelor clinice n perioada 2001-2005, se constat o
valoare medie de 8,0%, limitele de variaie fiind cuprinse ntre 6,4% (2003) i
10,8% (2004), (fig. 3).
Rezultatele obinute sunt similare cu cele ale altor autori (7, 9).

Fig. 3 Dinamica incidenei mamitelor clinice la vacile cu lapte (2001-2005)

n anul 2005, incidena mamitelor clinice n efectivul matc de 460 capete a


fost de 32,2% (148 vaci), repartiia pe sezoane fiind de 96 vaci (64,8%) n cel cald
i de 52 vaci (35,2%) n cel rece. n urma tratamentelor aplicate cu antibiotice
94,6% (140 vaci) s-au vindecat, iar 6,8% (8 vaci) au fost sacrificate.

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Studiind incidena mamitelor clinice la vaci n funcie de rangul lactaiei


s-au constatat valori maxime la a-IV-a i a-V-a lactaie (23,7%), perioad care
coincide cu vrful lactaiei, cnd vacile ajung pn la 8000 l / lactaie normal i
valori minime la a-VI-a lactaie (5,3%), cnd curba lactaiei este n scdere (fig.
4). Aceast situaie a fost constatat i de ali autori (9). Astfel, se confirm faptul
c vacile cu producii mari de lapte de peste 5000- 6000 l / lactaie normal
prezint o inciden a mamitelor clinice mai mare comparativ cu vacile care au
producii mai mici.

Fig. 4 Dinamica incidenei mamitelor clinice la vaci n funcie de rangul lactaiei


(2001-2005).

n funcie de anotimpuri incidena mamitelor clinice a nregistrat valori


minime iarna (17%) i valori maxime primvara (38,0%). Aceast situaie se
explic prin faptul c, spre sfritul iernii i nceputul primverii se nregistreaz
un vrf al ftrilor, cnd se manifest i o scdere a reaciilor de autoaprare n
perioada postpartum, cnd femelele prezint o susceptibilitate crescut la infecii
pe fondul unei imuniti sczute, dup o perioad de hrnire cu furaje din stoc,
furaje conservate, deficitare n principii vitamino-minerali (fig. 5).

Fig. 5 Incidena mamitelor clinice n funcie de anotimpuri (2001-2005)

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Fig. 6 Incidena mamitelor clinice n funcie de sezonul de stabulaie (2001-2005).

n funcie de sezon incidena mamitelor clinice a prezentat valori maxime


n sezonul de stabulaie de primvar-var (62%) i valori minime n cel de
toamn- iarn (38%), (fig. 6).
n sezonul de var temperaturilor caniculare i ventilaia necorespunztoare
din adposturile cu vaci acioneaz ca factori de stress scznd rezistena la
infecii .
Factorii tehnologici, cu referire la lungimea patului la animale, modul de
evacuare a dejectiilor din adpost, felul aternutului (paie, covor de cauciuc, nisip)
trebuie luai n considerare n programul general de prevenire a mbolnvirii
animalelor, deoarece ei reprezint de asemenea factori importani care
influeneaz starea de sntate a glandei mamare i salubritatea laptelui obinut,
fapt constatat i de alti autori (4, 5, 6).
Pe lng efectele negative asupra produciei de lapte, vacile cu mamite
clinice prezint o rat de concepie sczut i o rat crescut a avorturilor, ceea
ce afecteaz funcia de reproducie a femelelor cu repercusiuni negative asupra
indicilor de producie i eficien economic a fermei (8).
n condiiile n care incidena mamitelor n ferma de vaci cu lapte prezint
valori ridicate, care variaz n funcie de diveri factori, se impun msuri de
prevenire a acestor afeciuni, cu referire la modernizarea tehnologiilor de
ntreinere, exploatare, furajare a animalelor, mbuntirea ventilaiei n
adposturi, igiena glandei mamare, igiena mulsului, nrcarea corect a vacilor
gestante n ultimele dou luni, nainte de ftare, aplicarea de tratamente de
prevenire a infeciilor mamare n perioada de repaus mamar, tratarea corect i
rapid a cazurilor clinice i reformarea cazurilor cronice, care recidiveaz,
stopnd rspndirea bolii n efectivul de animale.

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CONCLUZII

1. Incidena mamitelor clinice n perioada 2001-2005 a fost cuprins ntre


6,4% (2003) i 10,8% (2004), media multianual fiind de 8,0% ;
2. Mamitele clinice la vacile n lactaie sunt pe locul patru, cu o medie de
8% n clasamentul cauzelor de ieiri din efectiv prin sacrificare, dup afeciunile
aparatului genital (37,4%), afeciunile membrelor (37,5%) i afeciunile medicale
(15,1%);
3. Incidena mamitelor clinice raportat la rangul lactaiei a prezentat valori
maxime de 23,7% la lactaiile IV-V i valori minime la a VI-a lactaie (5,3%);
4. n funcie de anotimp, incidena mamitelor clinice nregistreaz valori
maxime primvara (38%) i valori minime iarna (17%), iar n funcie de sezon,
valori maxime n sezonul primvar var (62%) i valori minime n sezonul
toamn- iarn (38%);
5. Pentru a reduce pierderile cauzate de manifestarea mamitelor la vacile n
lactaie se impune aplicarea unui program complex de msuri de prevenire,
monitorizare a strii de sntate a glandei mamare, igien a mulsului, exploatare
i furajare corespunztoare, nrcarea corect a vacilor gestante i reformarea
cazurilor de mamit cronic.

BIBLIOGRAFIE

1. Cengiz, Yalin, 1999 - Cost of Mastitis in Scottish Dairy Herds with Low and High
Subclinical Mastitis Problems - Department of Livestock Economics, Faculty of
Veterinary Science;
2. Hillerton J.E., E.A. Berry, 2005 - Treating mastitis in the cow a tradition or an
archaism- Journal of Applied Microbiology- Vol. 98, Pg 1250, England;
3. Hurley W.L, D.E Mori, 2000 Mastitis - Lactation Biology, Illinois, USA;
4. John H. Kirk, 1999 - Principle- based mastitis prevention - Nature Biotechnology, 70
DVM, MPVM, Veterinary Medicine, Extension, University of California- Davis,
Veterinary Medical Teaching and Research Center, Tulare, CA, USA;
5. Krznari, Marko, 1995 - Evaluation of damage caused by mastitis in cows -
SVIBOR - Veterinarski Institut, Zagreb (48);
6. Rosca P., L. Runceanu, D. Drugociu, Liliana Rosca, 2002 - Clinical mastitis dairy
cows: Frequency and variations in function of season and breeding condition -
Lucrri tiintifice, vol. 45(4), Edit. Ion Ionescu de la Brad, Iasi, pg. 422-426;
7. Rosca P., L. Runceanu, D. Drugociu, Liliana Rosca, 2002 - Researches regarding
the correlation between the frequency of clinical mastitis and level of milk-
production at dairy cows - Lucrri tiintifice, vol.45(4), Edit. Ion Ionescu de la
Brad, Iasi, pg. 418-421;
8. Santos J.E.P., R.L.A. Cerri, M.A. Ballou, G. Higginbotham, J.H. Kirk, S.O.
Juchem, 2001 - Effect of clinical mastitis incidence on lactational and
reproductive performance of Holstein dairy cows. - J. Anim. Sci. Vol. 80, Suppl.
1/J. Dairy Sci. Vol. 85, Suppl. 1 373, Sherbrooke University, Quebec, Canada;
9. Winston Ingalls, 2003 - Clinical Mastitis - West Agro, Inc., Aug 2, Kansas City,USA.

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STUDII PRIVIND INCIDENA MAMITELOR SUBCLINICE


LA VACI N FUNCIE DE UNII FACTORI DE VARIAIE
THE STADIES REGARDING THE INCIDENCE OF SUBCLINICAL MASTITIS
AT DAIRY COWS IN FUNCTION OF SOME VARIATION FACTORS

N. HROMEI
A.N.S.V.S.A. - PIF Sculeni
Elena RUGINOSU, Mariana SOFRONIE,
M. PNTEA, C. BDELI, Anca LUPU
S.C.D.B. DANCU Iasi

The researches were made into a dairy farm, Black and White
Spoted breed, exploited in an intensive system, the milk production being of
6700 milk l.cow/ year. The objective of the study was to establish the
dynamic incidence of the subclinical mastitis according to some variation
factors: season, lactation range and milk production /day.
In this purpose were taken samples of milk from 90 lactation cows,
which were tested with a reactive R. Mastitest in a fast stable test and from
the interpretation of the date (positive and doubtful test) was established
the state of udder health regarding the incidence of subclinical mastitis.
The results obtained were indicated the following variations of the
values regarding the incidence of subclinical mastitis at cows: the higher
values with 11,7% on the posteriore mammare quartiers, comparative with
the anteriore mammare quartiers (44,6% and 32,9%, respectively) and with
3,2% on the left mammare quartiers, comparative with the right side of the
udder (40,3% and 37,1%, respectively); higher values with 19,4% in the
summer, comparative with the winter season, significantly difference (P <
0,05), (54,54 and 35,07%, respectively); in raport with lactation range,
smaller values with 14,1%, 16,7% and 19,9% at the first lactation (24,4%),
comparative with the following lactations: the second, the third and the four
lactation (38,5% , 41,1% and 44,3% , significantly differences (P < 0,05);
in raport with milk production were smaller values with 7,1% and 26,7%
(significantly differences, P < 0,01) at the cows with 10-20 l. milk/day
(31,2%), comparative with the cows with 21-30 l. milk/ day (38,3% and
respective 57,9% ).

Key Words: dairy cows, R. Mastitest, subclinical mastitis, incidence,


variation factors.

Mamitele la vacile cu lapte sunt cele mai importante afeciuni ale glandei
mamare cu implicaii zooeconomice deosebite, fiind dintre cele mai pgubitoare
afeciuni.

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S-a constatat c n efectivele de vaci cu mamit se nregistreaz pierderi n


producia de lapte estimate la 100-140 $ / vac, ceea ce reprezint 70% din costul
total al mamitelor (costuri cu medicamentele, fora de munc suplimentar,
serviciile veterinare, eliminarea laptelui dup tratamentele cu antibiotice, scderea
calitii laptelui i n consecin preuri mai mici ale acestuia, reformarea timpurie
a animalelor). Din totalul costurilor, 10-26% sunt datorate mamitelor subclinice
(5).
Lund n considerare aceste aspecte economice, dar i cele ce privesc
biosecuritatea consumatorilor umani, pstrarea strii de sntate a glandei mamare
la vacile cu lapte trebuie s fie un obiectiv important al programului general de
prevenire i supraveghere a sntii animalelor din ferm, condiie pentru
asigurarea eficienei fermei i salubritatea produselor obinute. Respectarea unei
igiene corespunztoare a mulsului, asigurarea de condiii optime de microclimat
n adposturi, controlul periodic al parametrilor de funcionalitate a aparatelor de
muls, supravegherea strii metabolice i imunologice a animalelor, un sistem
mbuntit de ntreinere, furajare i exploatare a vacilor pentru lapte, precum i
controlul periodic al mamitelor subclinice, tratarea preventiv la nrcare i
tratarea formelor clinice de mamit n perioada de lactaie sunt cteva din
aspectele ce privesc acest program complex de prevenire a mamitelor (2, 4, 6).
Numeroasele studii efectuate au evideniat o frecven variabil a
mamitelor subclinice la vacile cu lapte, datorit complexitii i diversitii
factorilor implicai n etiopatogenia acestei entiti patologice i deseori datorit
unei slabe eficiene a msurilor de prevenire i tratament (1, 2, 7, 8).
Scopul studiilor a fost acela de a stabili dinamica incidenei mamitelor
subclinice la vacile cu lapte n funcie de unii factori de variaie: sezon, numrul
lactaiei i producia de lapte/ vac /zi.

MATERIAL I METOD DE LUCRU

Cercetrile au fost efectuate n perioada ianuarie 2004 - decembrie 2005,


n ferma de cretere i exploatare a vacilor cu lapte SCDB Dancu Iasi, indemn
de boli infecto-contagioase, LEB i TBC.
Materialul de studiu a fost reprezentat de vaci, ras Blat cu Negru
Romneasc, la care efectivul matc a fost n medie de 400 vaci cu lapte.
Sistemul de exploatare a fost de stabulaie permanent, cu excepia
sezonului cald cnd s-a practicat un sistem mixt, cu meninerea animalelor n
tabara de var, n sistem liber, cu padoc de pmnt, cu front de furajare betonat
i prevzut cu umbrar.
Furajarea animalelor s-a fcut cu furaje obinute n propria ferm i cu
reziduri industriale provenite de la fabricile F.N.C. i fabrica de vin-alcool
(borhoturi de bere). Raia furajer a fost alctuit difereniat n funcie de starea
fiziologic a animalelor, producia de lapte, sezon, vrst.
Mulsul vacilor s-a efectuat mecanic cu instalaii la bidon, tip Banat, de
dou ori pe zi (dimineaa i seara). Repausul mamar de dou luni nainte de

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ftare a fost respectat cu strictee pentru refacerea rezervelor corporale a


animalelor i susinerea viitoarei lactaii la parametrii ridicai. Producia medie de
lapte realizat pe lactaie normal a fost de 6700 l. / vac.
n vederea depistrii mamitelor subclinice la vacile n lactaie s-au aplicat
teste rapide de grajd cu ajutorul reactivului R. Mastitest produs de Institutul
Pasteur Bucureti). S-a stabilit incidena mamitelor subclinice n funcie de unii
factori de variaie, cum ar fi sezonul, rangul lactaiei, producia de lapte, precum i
repartiia afeciunii pe sferturile mamare.
Interpretarea rezultatelor s-a fcut dup aspectul amestecului lapte -
reactiv R. Mastitest (1):
- reacie negativ (-) = amestecul rmne fluid, omogen i subire ;
- reacie dubioas () = n amestec apar filamente i flocoane;
- reacie pozitiv (+) = amestec uor consistent, cu flocoane i fluiditate
sczut;
- reactie intens pozitiv (+ +) = amestec foarte consistent i vscos.
Dac se nclin placa amestecul rmne prins de plac sau se scurge
ca un lichid filant.
Rezultatele au fost prelucrate statistic i au fost prezentate n grafice i
tabele.

REZULTATE I DISCUII

Rezultatele investigaiilor efectuate n cadrul acestui studiu referitoare la


incidena mamitelor subclinice, prezentate sintetic n tabele i grafice evideniaz
unele variaii n raport cu diferii factori. Analiza datelor indic o inciden a
mamitelor subclinice mai mare cu 11,7% la sferturile situate n partea posterioar
a glandei mamare, comparativ cu cele din partea anterioar (44,6% fa de 32,9%)
i cu 3,2% la sferturile din partea stng, comparativ cu cele din partea dreapt a
ugerului (40,3% fa de 37,1%), (tab. 1a, tab. 1b, fig. 1). Rezultate similare au
fost evideniate i de ali autori (7).

Partea anterioar posterioar dreapta stnga

Fig. 1 Incidena mamitelor subclinice la vaci pe sferturi mamare

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n raport cu sezonul incidena mamitelor subclinice la vaci nregistreaz


valori cu 19,4% mai mari n sezonul de var, comparativ cu cele din iarn,
diferena fiind semnificativ (P < 0,05), (54,54% fa de 35,07%), (tab. 2, fig. 2).

Tabelul 2
Rezultate privind manifestarea mamitelor subclinice la vaci n funcie de sezon

Sezonul
Interpretarea reactiei R. Total probe
Mastitest Var Iarn
n % n % n
Probe negative ( - ) 30 45,50 187 64,90 217
Total probe mamit subclinic
36 54,54 101 35,07 137
( ) si ( + ) din care:
- dubioase ( ) 17 25,80 74 25,70 91
- pozitive ( + ) 10 15,20 17 5,90 27
- intens pozitive ( ++ ) 9 13,60 10 3,50 19
Total sferturi mamare testate 66 288 354

Fig. 2 Dinamica incidenei mamitelor subclinice la vaci n funcie de sezon

Analiznd manifestarea mamitelor subclinice n funcie de numrul lactaiei


se constat valori mai mici cu 14,1%, 16,7% i 19,9% la prima lactaie (24,4%),
comparativ cu lactaiile a-II-a, a-III-a i a-IV-a (38,5%, 41,1% i respectiv
44,3%), diferenele fiind semnificative (P< 0,05), (tab.3, fig. 3).

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Tabelul 3
Rezultate privind manifestarea mamitelor subclinice la vaci
n funcie de numrul lactaiei.

Numrul lactaiei
Total
Interpretarea a-IV-a i probe
reaciei R. Mastitest I-a a-II-a a-III-a
peste a-IV-a
n % n % n % n % n
Probe negative ( - ) 31 75,6 96 61,5 46 58,9 44 55,7 217
Total probe
mamit subclinic 10 24,4 60 38,5 32 41,1 35 44,3 137
( ) si ( + ), din care:
- dubioase ( ) 10 24,4 38 24,4 24 30,7 19 24,1 91
- pozitiv ( + ) - - 12 7,7 - - 15 18,9 27
- intens pozitiv ( ++ ) - - 10 6,4 8 10,2 1 1,3 19
Total sferturi
41 156 78 79 354
mamare testate

Fig. 3 Dinamica incidenei mamitelor subclinice la vaci n funcie de numrul lactaiei

n funcie de producia de lapte /zi se constat valori ale incidenei


mamitelor subclinice mai mici cu 7,1% la vacile cu producii de 10 - 20 l / zi
(31,2%), comparativ cu vacile cu producii de 21- 30 l/ zi (38,3%) i mai mici,
respectiv cu 26,7% (diferen distinct semnificativ, P < 0,01 ) fa de vacile cu
producii de 31-35 l/ zi (57,9%), (tab.4, fig. 4).
Literatura de specialitate indic faptul c, probele pozitive la testele de
control (R.Mastitest, CMT), ct i probele dubioase pun n eviden creteri ale
numrului de celule somatice peste limitele permise, precum i a numrului total
de germeni din lapte, ceea ce indic existena unei infecii la nivelul glandei
mamare (7).
Analiza rezultatelor din acest studiu prezint faptul c, incidena mamitelor
subclinice variaz n funcie de unii factori, care acioneaz ca supresori ai
sistemului imunitar scznd rezistena organismului la diferii ageni patogeni din
mediul extern sau intern. Astfel, n sezonul de var temperaturile caniculare au

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indicat un efect negativ asupra strii de sntate n general i a glandei mamare, n


mod special, fapt ce a determinat o cretere a susceptibilitii la diferite infecii.
Acest fapt a fost confirmat i de ali autori (6, 7).
Tabelul 4
Rezultate privind manifestarea mamitelor subclinice la vaci
n funcie de producia de lapte

Producia de lapte
Total
Interpretarea reaciei l./ zi
probe
R.Mastitest 10 - 20 21 - 30 31 - 35
n % n % n % n
Probe negative ( - ) 42 69,8 169 61,6 8 42,1 217
Total probe mamit subclinic
19 31,2 105 38,3 11 57,9 137
( ) si ( + ), din care:
- dubioase ( ) 12 19,7 74 27,0 7 36,8 91
- pozitive ( + ) 5 8,2 15 5,5 3 15,8 27
- intens pozitive ( ++ ) 2 3,3 16 5,8 1 5,3 19
Total sferturi mamare testate 61 274 19 354

Fig. 4. Dinamica incidenei mamitelor subclinice la vaci n funcie de producia de lapte/zi.

Factorii de zooigien a mamelei au un rol deosebit de important n


asigurarea sntii glandei mamare. O deficien n aceast direcie i mai ales la
nivelul posterior a sferturilor mamare are efecte negative, crescnd incidena
mamitelor, aa cum s-a constatat n studiul de fa i cum a fost confirmat i de
ali autori (7).
Datele nregistrate n acest studiu indic faptul c, vacile cu producii
ridicate de lapte aflate n faza de vrf a lactaiei sunt mai susceptibile la infecii,
datorit metabolismului lor mai intens i a necesarului crescut de nutrieni energo-
proteici i vitamino-mineral, care n anumite situaii este greu de asigurat la
nivelul cerinelor. La vacile cu mai multe lactaii (peste a- IV-a) s-a constatat de
asemenea o inciden crescut a mamitelor datorit scderii sistemului imunitar

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pe fondul unor deficiene metabolice i a predispozitiei spre mamite prin


recidivarea unor infecii anterioare.
n vederea reducerii incidenei mamitelor n general i a celor subclinice n
mod special se impune aplicarea unui program de supraveghere permanent a
strii de sntate a glandei mamare, n aceast direcie acest test rapid de grajd cu
reactiv R. Mastitest poate fi de un real ajutor.
Cunoscnd factorii de variaie a acestor afeciuni se poate interveni n
sensul minimalizrii aciunii lor, contribuind la reducerea pierderilor economice i
la creterea eficienei exploatrii vacilor cu lapte.

CONCLUZII

1. n funcie de repartiia pe sferturile mamare s-a constatat inciden a


mamitelor subclinice mai mare cu 11,7% la sferturile situate n partea posterioar
a glandei mamare, comparativ cu cele din partea anterioar (44,6% fa de 32,9%)
i cu 3,2% la sferturile din partea stng, comparativ cu cele din partea dreapt a
ugerului (40,3% fa de 37,1%);
2. n raport cu sezonul incidena mamitelor subclinice la vaci nregistreaz
valori cu 19,4% mai mari n sezonul de var, comparativ cu cele din iarn,
diferena fiind semnificativ (P < 0,05), (54,54% fa de 35,07%);
3. n funcie de numrul lactaiei se constat valori mai mici cu 14,1%,
16,7% i 19,9% la prima lactaie (24,4%), comparativ cu lactaiile a-II-a, a-III-a i
a-IV-a (38,5%, 41,1% i respectiv 44,3%, diferenele fiind semnificative (P<
0,05)
4. n funcie de producia de lapte /zi se constat valori ale incidenei
mamitelor subclinice mai mici cu 7,1% i respectiv cu 26,7% (diferene distinct
semnificative, P < 0,01) la vacile cu producii cuprinse ntre 10-20 litri lapte/ zi
(31,2%), comparativ cu vacile cu producii de 21-30 l. lapte/ zi i respectiv 31-35
l. lapte / zi (38,3% i respectiv 57,9%)

BIBLIOGRAFIE

1. Broiu A., 2002 - Diagnosticul mamitelor la vacile de lapte- Buletin informativ ,vol. I,
nr. 2;
2. Gheorghiu Anna, Statov C., Harmat Eugenia, 1979 - Calitatea igienic a laptelui n
raport cu sistemul de evacuare a dejeciilor i tipul de aternut din adposturile
de vaci de lapte- Lucr. St. Taurine, ICDB Balotesti, pg. 119- 125;
3. Groza I., Bogdan L., Simona Ciupe, R. Pop, Moser F., Cenariu M, Trenisan T.,
Filip D., 2004 - Cercetri privind incidena i tratamentul mamitelor subclinice i
clinice la vac - Lucrari stiintifice, USAMV, Seria C., vol. XLVI-XLVII, Facultatea
de Medicina Veterinara Cluj-Napoca;
4. Jones G.M., T.L. Bailey, Jr., 1998 - Mastitis Control in Heifers and First Lactation -
Maryland College of Veterinary Medicine, and Extension Veterinarian, Virginia
Tech Publication Number 404-281
5. Hurley W.L, D.E Mori, 2000 Mastitis - Lactation Biology, Illinois, USA

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Universitatea de tiine Agricole i Medicin Veterinar Iai

6. Ken E. Leslie, 1997 - Fine-Tuning Mastitis Control Programs Department of


Population Medicine, University of Guelph, Guelph, Ontario, Canada N1G 2W;
7. Kikkers B.H., L. zsvri, F.J.C.M. Van Eerdenburg, L. Brzsnyi, O. Szenci,
2004 - The incidence of mastitis treated with antibiotics in large- scale Hungarian
Holstein-Frisian dairy farms - Acta Veterinaria Hungarica 52 (1), pp. 1932;
8. Mi McFadden, 1996 - Influence of Dry Cow Management on Mastitis Michigan Dairy
Review 1(4):12, USA.

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HOOP STRUCTURES DESIGN FOR GESTATING SOWS:


REVIEW
STRUCTURI UOARE PENTRU NTREINEREA SCROAFELOR GESTANTE:
RECENZIE

I. HUTU*
U.S.A.M.V.B. Timioara

Structurile uoare semicirculare pot fi alternative ieftine, practice i


aplicabile n sectorul creterii suinelor din Romnia. Structurile hoop se
caracterizeaz prin investiii reduse, ntreinere n grupuri i pe aternut
permanent. Conform citrilor bibliografice parametri bunstrii, unele
caractere productive i reproductive sunt, n bun msur, superioare sau
comparabile altor sisteme de producie.
Scopul acestei lucrri este derularea unui parteneriat cercettor
extensionist - fermieri i nceperea cercetrii, extensiei i practicii
productive n astfel de strcucturi.

Key words: hoop, structures, gestation and, sows

The hoops, which originated in the prairie provinces of Canada, are low
cost and easy to erect. They rely on the use of large amounts of bedding to absorb
the pig feces and urine and also for the pig sleeping areas. During the past fifteen
years, hoop structures or hoops have become a popular alternative for housing
finishing pigs in Australia, Canada, Korea, Nederland, Ukraine and USA. Hoops
are large tentlike structures that are deep bedded with cornstalks or straw. [3, 6, 7]
For Romanian transition period hoop structures can be a cheaper alternative at
expensive conventional structures.
In 2005, February 8th in the Agroinfo Forum (http://forum.agroinfo.ro/) was
start hoop structure topic and till 2006 February 8th 63 replies and 21.812 views
was done. [11] We believe that Romanians farmers' attitudes will be open for low-
input technology as well as other farmers. [9]
The aims of this paper it to start on a partnership relation researcher-
extensionit-farmers in order to promote and use these structures.

*
E-mail: office@unitate-extensie.org.ro

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MATERIALS AND SYSTEM DESIGN OF HOOP


FOR GESTATING SOWS

Hoop structures, or hoops, consist of arched pipes or trusses in a Quonset-


shaped structure covered with an ultraviolet-resistant polyethylene fabric tarp.
The hoops are attached to wooden posts with wood sidewalls that are 1.22 m
high. The basic particularities of hoop are:
- the system uses group housing of sows in deep straw during breeding
and/or gestation.
- the stalls have front access gates that open into the feeding alley.
- the group housing helps stimulate estrus and reduce stress on the
sows by allowing them to live in groups, which they prefer.
- fighting is minimized by using feeding stalls and introducing new sows
to the groups at optimum times, such as at farrowing.
Examples of design: Pen layout is dependent on feeding system. Several
layouts are shown by Honeyman and all (1997). The gestation hoops sown in
figure 1 are designed for two groups of 32 sows. Feed stalls are placed on a
raised (+0.40 cm) concrete pad at the end of the hoop. Feeding stalls are placed
so that only one group of sows at one time may be allowed to feed. This design
minimizes the equipment cost by allowing feeding stalls to be shared but some
space is wasted to accommodate the feeding stalls. Shared feeding stall usage
on a daily feeding system will require some added labor.

Fig. 1. Hoop housed gestation with shared feeding stalls for two groups of 32 sows
(Honeyman, Harmon, Lay and Richard, 1997, Iowa State University)

Figure 2 illustrates a relatively simple feeding stall system for 64 sows. This
system is efficient and simple. The main disadvantage is the cost of the feeding
crates.
Figure 3 shows four groups of 25 that share a feeding area in the center of
the building.

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This design minimizes the area devoted for feeding but animals will be
exposed to different conditions depending on where in the system they are
housed. Added labor may be required because more than one group may need to
be fed in a day.

Fig. 2. Hoop housed gestation with 64 feed stalls


(Honeyman, Harmon, Lay and Richard, 1997, Iowa State University)

Fig. 3. Hoop housed gestation with interval feeding for four groups of 25 sows
(Honeyman, Harmon, Lay and Richard, 1997, Iowa State University)

PRACTICAL RESULTS AND DISCUSSIONS

The performance efficiency of gestating and breeding pigs may be affected


by the management and environment in which they are housed.
Layout and management considerations: Depends of the type, the floor
plan includes lockable feeding stalls with front access doors, a feeding and service
alley and a deep bedded area for the group-housed sows. Number of sows per pen
or group is a factor of farrowing room/house capacity and conception rate. Pens
should be at least 4.60 or 4.90 m wide to minimize control by aggressive sows

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and allow timid sows to pass freely. All floor areas of the hoop structure are
concrete or partial concrete and partial soil. Most of the floor is earthen and
covered with bedding.
Swedish guidelines for bedded space of 2.25-2.50 m2/ gestating sow are
recommended.
Temperature. Reproduction efficiency of boars and sows is generally
unaffected by cold temperatures unless scrotal frost-bite occurs with boars or
gestating sow feeding level is too low. Sows and boars housed in colder
conditions will require additional feed. The recommendation is 0.33 kg of
additional feed per every 5C colder than 13C. Hoop structures are designed to
serve as cold housing. Heat is not added to the structure and the indoor air
temperature will generally remain 3 to 6C above the outdoor temperature.
Sows should be maintained with effective temperatures below a daily
average of 26C and below 30C as a maximum. To achieve this, less bedding
will be used and an interval sprayer or dripper system will probably be necessary.
This may prompt the sow to burrow into moist places in earthen floors, resulting
in a problem during clean-out.
Ventilation. The structure is naturally ventilated and drippers were installed
over the feeding stalls to aid in cooling the sows on hot days.
During summer months both ends of the hoop should be left open to allow
air flow through the structure.
During winter the north end of the hoop will generally remain closed. Air
comes in through slots on either side of the frame walls, rises to the top of the
hoop, and moves out the south end, which remains open. If the structure is closed
too tightly the humidity will build up and cause possible respiratory problems.
Sows remain comfortable in cold conditions because they can modify their
environment with the use of bedding and huddling with other animals. They will,
however, require more feed because they will use more energy to maintain
homeothermic conditions. A high quality bedding source is essential to the
success of this structure.
Feeding and sow management. Usually, the feeding stalls and energy-free
waterers are place on a raised (+40 45 cm) concrete platform. During gestation
the sows are fed daily in individual feeding stalls with rear gates that can be
closed after the sows have entered the stalls. The feeding stalls offer the
advantages of individual gestation crates: individual feeding, control of the sow
for vaccinations and artificial insemination, and easy sorting of sows. The group
housing helps stimulate estrus and reduce stress on the sows by allowing them to
live in groups. Fighting is minimized by using feeding stalls and introducing new
sows to the groups at optimum times, such as at farrowing.
Bedding and manure management. Bedding can be cornstalks, straw, wood
shavings, or other absorbent organic material. Materials (kg per sow turn) are:

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corn stalks (300 to 450), barley straw (360 to 1200), oat straw (600 to 550), wheat
straw (340 to 510) or pine shavings (380 to 570). [1, 2]
Animals bed and dung on a bedded manure pack, which generates heat and
contributes to animal comfort during winter months. Access for adding bedding is
from the south end. The north end is closed in winter with a fabric curtain. The
south end is partially closed in winter. Large, round bales of straw or cornstalks
can be used for bedding. The bedded-pack is normally cleaned out after each
group of sows is sent to farrowing house, 3 to 4 times a year. [8]
Initial investment cost. In hoop 2.20-2.50 m2 per sow and in confinement
crate 1.30-1.50m2. In those cases the cost for on sow in gestation sector is $180 to
$325 per sow in hoop structures and $300 to $550 per sow in confinement crate.
The confinement facilities are depreciated over 16 years (6.25% annually), while
the hoops are at 10 years (10% annually). [5]
The reproductive performances in comparison hoops vs. individual
gestation crates are comparable. Lammers and Honeyman (2004) communicated
preliminary trends 0.7 more pigs per liter (10.9 vs. 10.1) and lower sows removal
from hoop structures (7% vs. 14%). In conclusion the conception rates are high,
sow longevity is long, and sow mortality and culling rates are low.
Animal welfare. Weng et al., (1998) concluded that in group housed sows
and floor space a minimum of 2.4 to 3.6 m2 required promoting good welfare.
It was demonstrated that hoops sows: performed fewer abnormal behaviors,
had lower plasma cortisol concentrations, greater rate of play behavior and fewer
leg injuries. Thus indicating they had better welfare.

CONCLUSIONS

The hoops have the advantages of low initial cost, versatility, and
simplicity.
Hoops offer an alternative to conventional confinement housing systems.
Hoops are a sustainable alternative for producing pigs because they are 1)
environment-friendly with solid manure that can be composted and, 2) farmer-
friendly with low capital cost and competitive cost of production.
The Romanian family farmer can compete and be a viable entity in the
Romanian swine industry

REFERENCES

1. Honeyman, M.S., Harmon, J.D., Kliebenstein, J.B., Richard, T.L., 2001 -


Feasibility of hoop structures for market swine in Iowa: Pig Performance, Pig
Environment, and Budget Analysis. Applied Engineering in Agriculture
17(6):869874.
2. Honeyman, M.S., Kent, D, - Reproductive Performance of Young Sows from Various
Gestation Housing Systems In: 1999 ISU Swine Research Report. ASL-R1678
Iowa State University, Ames Iowa State University Management/Economics.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

3. Honeyman, M.S., Harmon. J., Lay, D., Richard, T. - Gestating Sows in Deep-
bedded Hoop Structures ASL-R1496 Management/Economics Iowa State In:
1997 ISU Swine Research Report. AS-638. Dept. of Animal Science, Iowa State
University, Ames
4. Lammers, P., Honeyman, M. - Effects of bedded gestation housing on litter size and
culling rates: preliminary results National Conference of Hoop Barns and Bedded
th
Systems for Livestock Production. September 14 2004, Ames, Iowa State
University, abstract
5. Lawrence, J., Vontalge, A. - Livestock Enterprise Budgets for Iowa - 1998. Iowa
State University, University Extension, Ames.
6. Maltman J. - Manitoba Agriculture, Food and Rural Initiative. Hoop barns in Canada.
National Conference of Hoop Barns and Bedded Systems for Livestock
Production. September 14th 2004, Ames, Iowa State University, presentation
7. Richard, T.L. - Alternative swine housing opportunities for innovation. National
Conference of Hoop Barns and Bedded Systems for Livestock Production.
th
September 14 2004, Ames, Iowa State University, abstract
8. Richard, T.L., Harmon, J., Honeyman, M., Creswell, J., 1998 - Hoop Structure
Bedding Use, Labor, Bedding Pack Temperature, Manure Nutrient Content, and
Nitrogen Leaching Potential. ASLR1499. In: 1997 ISU Swine Research Report.
AS-638. Dept. of Animal Science, Iowa State University, Ames, IA. pp 71-75
9. Sharp, Julie T., Hinrichs, C.C., 2001 - Farmer support for publicly funded
sustainable agriculture research: The case of hoop structures for swine.
American Journal of Alternative Agriculture 16(2): 81-88.
10. Weng, R.C., Edwards, S.A., English, P.R., 1998 - Behaviour, social interactions and
lesion scores of group-housed sows in relation to floor space allowance. Appl.
Anim. Behav. Sci. 59 4, 307-316
11. http://forum.agroinfo.ro/

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CORELAIILE EVIDENIATE NTRE NIVELUL


PRODUCIEI DE LAPTE, PERFORMANELE
REPRODUCTIVE I NIVELURILE SERICE ALE UNOR
HORMONI N PUERPERIUMUL VACILOR DE LAPTE DIN
RASA BLAT CU NEGRU ROMNEASC
CORRELATIONS BETWEEN MILK YIELD, REPRODUCTION AND HORMONAL
STATUS DURING POST PARTUM PERIOD IN ROMANIAN BLACK SPOTTED
DAIRY COWS

C. MIRCU, H. CERNESCU, G. OTAV,


Ilinca FRUNZ, Timea CZAPP,
M. MOISII, A. ARDELEAN
U.S.A.M.V.B. Timioara

First ovulation and pregnancy rate at first service are influenced by


a lot of factors during puerperal period of lactating dairy cows. On medium
milk yielding (G1, n=10) and high yielding (G2 n=10) Romanian Black
Spotted dairy cows we followed serum progesterone, thyroid homones (T3
and T4), glucose,insulin,IGF1 variations and BCS loss during first 60 days
and corellations to reproductive performance. We found that progesterone
and thyroid hormones levels arround insemination are crucial for embryo
survival and low T3 and T4 after 45 post partum days exert a negative
influence on pregnancy rate subsequent first insemination. We also found a
constant serum glucose and insulin level rises during puerperal, compared
to parturition moment, in both groups and also that cows with high milk
yield displays IGF1 serum values lower than values registered in medium
milk yielding cows. We concluded that in Romanian Black Spotted dairy
cows, irrelevant to milk yield, thyroid hormones, glucose, insulin and IGF1
levels, but no BCS loss do influence reproductive performances during post
partum period.

Pe parcursul perioadei puerperale, rezervele energetice ale organismului


matern trebuie s satisfac att cerinele produciei de lapte ct i pe cele ale
instalrii unei noi gestaii. Creterea necesarului energetic pentru producia de
lapte ar putea genera un bilan energetic negativ de-a lungul primelor sptmni
post partum, conducnd la tulburarea funciei de reproducie. Au fost evideniate
relaiile existente ntre nivelul produciei de lapte, bilanul energetic, aprecierea
conformaiei corporale (BCS) i rata gestaiei la prima inseminare, fiind de
asemenea identificai factorii care influeneaz n mod direct aceste procese
(ritmul eliberrii LH, nivelurile glicemiei, insulinei, IGF1, progesteronului i
hormonilor tiroidieni). Glucoza reprezint o surs major de energie pentru ovar
(Beam i Butler, 1999), n timp ce insulina influeneaz secreia GnRH i a LH

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(Butler, 2000). Efectele exercitate de IGF1 se regsesc n procesul dezvoltrii


foliculare (Lucy, 2000) i n cel al steroidogenezei (Whates i col., 2003). Dei
hormonilor tiroidieni li s-a relevat implicarea n termoreglare i homeostazia
metabolismului energetic, studii recente au evideniat implicarea lor n reglarea
funciei ovariene la rumegtoare, cu precdere la vacile aflate n puerperium
(Huszenicza i col., 2002). Reluarea ciclicitii sexuale post partum este nsoit
de creterea corespunztoare a progesteronului seric, nivelul atins de acest
hormon pe parcursul ciclului sexual fiind relevant pentru fertilitate (Lucy, 2001).
Evoluia bilanului energetic se reflect corespunztor n modificrile BCS, fiind
demonstrat relaia existent ntre bilanul energetic, BCS i fertilitate.
n studiul nostru am luat n considerare factorii relevani pentru
metabolismul energetic (glucoza, insulina, T3 i T4), pentru steroidogenez i
dezvoltarea folicular (IGF1 i progesteronul) i am urmrit modificrile BCS,
ncercnd s evideniem prezena unor relaii ntre aceti parametri i
performanele reproductive.

MATERIAL I METOD

n acest studiu desfurat n ferma de vaci a staiunii didactice a USAMVB


Timioara, n perioada aprilie octombrie 2005, au fost incluse 20 de vaci din
rasa Blat cu Negru Romneasc (BNR), aflate la a doua sau a treia ftare (n
medie 2,8). Femelele nu au avut antecedente obstetrical-ginecologice, au ftat
normal i nu au nregistrat retenia nvelitorilor fetale. Vacile au fost incluse n lotul
1 (L1, n = 10, vaci cu producie medie de lapte, 4172182,4 litri) i n lotul 2 (L2, n
= 10, vaci cu producie mare de lapte, 5418114,2 litri pe lactaie). Pentru
determinrile hormonale i ale glicemiei, probele de snge au fost recoltate la 7,
15, 30, 45 i 60 de zile post partum n ideea de a acoperi ntreaga perioad
puerperal. Pentru determinarea glicemiei i a hormonilor tiroidieni, au fost
recoltate probe de snge i la momentul parturiiei. La 20 de zile post partum a
debutat recoltarea sngelui, n vederea depistrii momentului primei creteri a
nivelului progesteronului. Probele de snge au fost recoltate prin puncia venei
jugulare, de dou ori pe sptmn (n zilele de luni i joi, dup mulsul de
diminea). Dup coagularea spontan la temperatura camerei, serul a fost
separat prin centrifugare (5 minute la 3000 rpm) i congelat (-20C) pn la
efectuarea determinrilor.
Nivelurile IGF1, progesteronului i hormonilor tiroidieni au fost determinate
prin tehnica ELISA, utiliznd truse de uz uman DRG Diagnostics (Germania),
truse care, opernd cu anticorpi policlonali, pot fi utilizate la analizarea serului
provenit de la vaci. Determinrile au fost executate n duplicat, citirea fcndu-se
pe cititorul ELISA PR 1100 (BIO RAD). Glicemia a fost determinat prin
intermediul metodei spectrometrice, cu ajutorul analizatorului VET SCREEN.
Valorile medii i deviaiile standard au fost calculate pentru fiecare lot, la fiecare
dintre momentele de referin anterior stabilite. Diferenele statistice au fost
stabilite prin intermediul testului ANOVA.

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Nivelurile insulinei au fost determinate cu ajutorul trusei DRG Diagnostics


(Germania) pentru insulin bovin. Pentru IGF1 a fost utilizat o trus de uz
uman, bazat pe anticorpi policlonali. Deoarece structura IGF1 este identic la
om, bovine i suine, n cazul utilizrii anticorpilor policlonali, procedeul este
validat pentru toate cele trei specii (Nikolic i col., 1996). Determinarea corect a
IGF1 presupune ndeprtarea proteinelor de cuplare a IGF1, datorit afinitii lor
similare pentru ligandul anticorpilor specifici utilizai n acest tip de reacie.
Probele au fost analizate dup o prealabil extracie cu acid-etanol, conform
metodei prezentate de Nikolic i col. (1996). Valorile BCS (conformaia corporal)
au fost estimate cu ajutorul metodei descrise de Ferguson i col. (1994) la
momentul parturiiei, i apoi la fiecare 15 zile, pn la 60 de zile postpartum.
ncepnd cu cea de-a 40-a zi post partum, vacile au fost atent urmrite
pentru depistarea estrului (de trei ori pe zi, cte 20 de minute). Vacile au fost
nsmnate la 12 ore de la depistarea strii de estru, urmrind regula AM/PM.
Femelele au fost urmrite n continuare, pentru a depista i utiliza estrul aprut n
cazul neinstalrii gestaiei. Diagnosticarea gestaiei s-a fcut prin examen
transrectal executat la 45-60 de zile de la nsmnare.

REZULTATE I DISCUII

Conform rezultatelor inserate n tabelul 1, prima cretere post partum a


progesteronului seric peste pragul de 1 ng/ml (nivel care indic prezena esutului
luteal i implicit ovulaia anterioar) a fost nregistrat la 29 de zile (pentru L1) i
la 37 de zile pentru L2, diferena fiind semnificativ (p<0,05). n cazul femelelor
din L1, ciclicitatea sexual a fost reluat cel trziu pn la cea de-a 34-a zi post
partum, iar pentru vacile lotului 2, nu mai trziu de ziua 41; de menionat c
niciuna dintre femele nu a manifestat anestru post partum.
Tabelul 1
Variaia nivelurilor serice ale progesteronului (XSx)

Prima cretere Nivelul mediu Nivelul mediu al


Prima IA (zile
Lotul nregistrat a P4 (zile al P4 dup 7 P4 la 7 zile post
post partum)
postpartum) zile (ng/ml) IA (ng/ml)
L1 29 2,38 3,8 0,35 46,6 3,11 5,11 1,37
L2 37,85 2,34 3,1 0,29 53 3,21 3,75 0,80

Staples i col. (1990) evideniaz prezena primei ovulaii la 22 de zile post


partum la vaci cu producii medii de lapte, n timp ce Opsomer i col. (2000)
semnaleaz prima cretere a nivelului P4 la 37 de zile post partum la vacile cu
producii mari de lapte.
La interval de o sptmn de la creterea iniial a nivelului P4, valorile
progesteronemiei nu au diferit semnificativ ntre cele dou loturi (3,8 ng/ml
pentru L 1, comparativ cu 3,1 ng/ml pentru L 2), situaie schimbat radical la 7
zile consecutiv nsmnrii artificiale, cnd diferena a fost semnificativ
p<0,01 - (5,11 ng/ml pentru L 1, fa de 3,75 ng/ml pentru L 2).

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Este interesant de menionat c dac, n cazul lotului 1, s-a evideniat o


diferen semnificativ (p<0,01) ntre nivelurile P4 luate n discuie (dup prima
ovulaie i consecutiv primei nsmnri artificiale), nu acelai lucru poate fi
observat n cazul lotului 2, unde concentraia seric a progesteronului nu a crescut
semnificativ. Acest aspect se coreleaz cu rata concepiei la prima IA, care este
complet nesatisfctoare la lotul 2. Meninerea gestaiei este condiionat i de
suportul progesteronic corespunztor, iar n cazul unor concentraii sczute ale P4
se poate presupune existena unor corpi luteali deficieni, a cror secreie nu poate
susine pstrarea gestaiei.
Capacitatea de a produce i menine concentraii optime ale P4 este
important pentru fertilitate datorit efectului reglator pe care acest hormon l
exercit asupra funciei endometrului; n zilele 4-7 post IA, P4 a fost crescut la
vacile care au rmas gestante, comparativ cu cele care nu au rmas (Lucy, 2001).
Este posibil ca nivelul relativ sczut al P4 n zilele 4-5 s permit recunoaterea
prezenei gestaiei i extinderea perioadei luteale, dar gestaia nu va progresa
deoarece embrionul a fost compromis pe parcursul dezvoltrii sale.
Dac n mod normal se observ o cretere progresiv a progesteronemiei
ntre primul i al doilea ciclu sexual post partum, precum i ntre al doilea i al
treilea (Staples i col., 1990), valorile medii obinute n prezentul studiu indic o
scdere progresiv a P4 ntre primul i al doilea ciclu n cazurile a dou femele
(25%) din L 1, i a dou femele (28,5%) din L 2. Trebuie menionat c aceste
femele au necesitat 3,5 nsmnri artificiale (L 1), respectiv 2,5 (L 2) pentru a
rmne gestante.
Cele dou femele mai sus menionate ale lotului 1 au nregistrat pe
parcursul primelor 60 de zile post partum niveluri ale T3 (tabelul 2) cuprinse ntre
0,2 i 0,8 ng/ml, iar n cazul celor dou femele din lotul 2 la care am fcut anterior
referire, valori cuprinse ntre 0,6 i 1,1 ng/ml. Chiar dac este normal ca n
primele sptmni consecutiv parturiiei vacile n lactaie s manifeste
hipotiroidism (Pezzi i col., 2003), studiul lui Jooritsma i col. (2003) a artat c
nivelurile serice ale T3 i T4 sunt mai sczute n cazul vacilor fr activitate
ovarian, precum i faptul c valori ale T3 situate sub 1,4 ng/ml sunt asociate cu
concentraii sczute ale estrogenilor i cu o slab exprimare a estrului. Analiznd
nivelurile T3, sesizm c n cazul femelelor aparinnd lotului 2, intervalul de
valori cuprinde concentraii mai crescute, ceea ce denot o eficient selecie a
vacilor pentru producia de lapte, cunoscndu-se faptul c declanarea secreiei
lactate este urmat de scderea nivelului T3.
Analiznd valorile incluse n tabelul 2, se evideniaz un hipotiroidism
marcant, persistent de-a lungul ntregii perioade considerate. Valorile medii ale
T3 sunt cele care ne atrag atenia, n cazul T4 scderea nefiind aa de accentuat.
Acest aspect este explicabil prin nsui mecanismul producerii T3 i al
intensificrii eliberrii rT3 n lapte (Pezzi i col., 2003).

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Tabelul 2
Variaia parametrilor investigai de-a lungul puerperiumului

Parturiie 7 zile 15 zile 30 zile 45 zile 60 zile


T3 (ng/ml)
L1 0,930,49 - 0,900,54 1,050,41 1,200,45 1,120,46
L2 1,010,36 - 0,780,34 0,800,45 1,140,44 1,240,32
T4 (ng/ml)
L1 25,753,69 - 21,36,93 27,06,23 26,875,20 28,258,86
L2 23,144,74 - 21,282,75 21,715,7 23,143,71 24,204,27
Glicemia (mmol/l)
L1 3.030.3 3.060.3 3.110.34 3.280.45 3.330.48 3.320.53
L2 2.670.3 2.600.3 2.800.37 2.920.28 2.950.49 2.950.43
Insulina (U/l)
L1 - 8.200.5 8.460.56 9.110.79 9.751.20 -
L2 - 7.871.0 8.251.19 9.181.06 10.051.64 -
IGF 1 (ng/ml)
L1 - 74.8311.43 81.6112.8 88.9812.59 94.4311.83 -
L2 - 68.179.06 71.4712.2 88.1012.47 94.9512.87 -
BCS
L1 3.030.1 - 2.870.18 2.780.16 2.620.23 2.650.26
L2 3.170.2 - 2.960.22 2.820.37 2.640.31 2.640.3

n cazul T3, nu exist o opinie unanim privind valorile minime i maxime


(Pezzi i col., 2003, 0,28-2,02 ng/ml; Pusta 2004, 0,2-2; Pechova i col., 2000,
0,68-1,68; Pavlata i col., 2004, 0,71-1,88), ns valoarea medie se situeaz n
jurul a 1,1 1,2 ng/ml.
Pentru tiroxin, valoarea minim admis pare a fi de 20 ng/ml (Pusta,
2004), regsind i n cazul acestui parametru diversitatea de opinii menionat
anterior (31,6 ng/ml Pezzi i col., 2003; 26,25 ng/ml Pechova i col., 2000;
33,2 ng/ml Pavlata i col., 2004).
Pe parcursul primelor 60 de zile post partum de-a lungul crora au fost
urmrite nivelurile T3 i T4, nu au fost evideniate diferene semnificative ntre
valorile medii ale celor dou loturi, ns a putut fi observat o corelaie ntre
nivelurile T3 i T4 la 45 de zile post partum i instalarea gestaiei consecutiv
primei nsmnri artificiale. Raportul T3:T4 a variat ntre 0,035 i 0,044 pentru
L1, i ntre 0,036 i 0,051 pentru L2, valori similare cu cele raportate de ali
autori. Astfel, Pusta (2004) nregistreaz valori ale raportului T3:T4 cuprinse
ntre 0,026 i 0,035, iar Pavlata i col. (2004), valori cuprinse ntre 0,039 i 0,055.
Dac pentru femelele lotului 1 rmase gestante n urma primei I.A., valoarea
medie a T3 la 45 de zile post partum a fost de 1,66 ng/ml, iar pentru cele din lotul
2 de 1,60 ng/ml, pentru cele care nu au rmas gestante consecutiv primei I.A.,
nivelul mediu al T3 la 45 de zile a fost de 0,92 ng/ml n cazul femelelor din L1 i
de 0,96 ng/ml pentru cele din L 2.

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Valorile glicemiei au variat n limite fiziologice (2,1 la 3,9 mmol/l) la toate


cazurile luate n studiu. La vacile lotului 1 se observ o cretere uniform
ncepnd cu parturiia i ajungnd la 60 de zile post partum (tabelul 2),
nregistrndu-se diferene semnificative (p>0,05) ntre valorile nregistrate la
momentul parturiiei i cele de la 30 i 60 de zile, dovendindu-se n acest mod
disponibilitatea organismului pentru neoglucogenez precum i o bine echilibrat
homeostazie a glucozei.
Valorile medii ale glicemiei (3,56 mmol/l) nregistrate la 60 de zile pp. la
acele vaci care au avut nevoie de 1-2 inseminri pentru a rmne gestante sunt
superioare celor care au necesitat 3 sau 4 inseminri pentru instalarea gestaiei.
Dac n cazul L1 doar dou vaci au necesitat 3 inseminri / gestaie, n cazul L2,
patru femele au avut nevoie de cte 3 inseminri pentru instalarea gestaiei.
Femela din L1 care a necesitat 4 inseminri pentru a rmne gestant a avut cea
mai sczut valoare a glicemiei (2,5 mmol/l) nregistrat pe parcursul ntregii
perioade de supraveghere, semnificnd necesitatea unei perioade mai lungi pentru
refacerea energiei ovariene disponibile.
Chiar dac variaiile s-au ncadrat n limitele fiziologice, vacile lotului L2
au nregistrat la parturiie niveluri ale glucozei semnificativ sczute , comparativ
cu cele ale lotului 1. La vacile L2, chiar dac glicemia a crescut pe msura
naintrii n puerperium, valoarea medie nregistrat la 60 de zile pp este mai
sczut dect valorile nregistrate de-a lungul ntregii perioade analizate pentru
L1.
Glucoza reprezint o surs major de energie pentru ovar, iar scderea
disponibilului de glucoz la nceputul lactaiei poate afecta negativ reluarea
activitii ovariene post partum (Rabiee i col., 1999). Nivelurile sczute ale
glicemiei n anumite momente post inseminare pot afecta cel puin parial
supravieuirea embrionului (Diskin i col., 2001). Din acest motiv am considerat
util urmrirea nivelului glucozei chiar i la 60 de zile pp., ns nu am gsit nicio
corelaie ntre valorile glicemiei i fecunditatea nregistrat la prima inseminare.
Concentraia plasmatic a glucozei poate fi utilizat n vederea estimrii statusului
energetic la vacile lactante, iar nivelurile sczute nregistrate de-a lungul
puerperiumului au fost corelate cu o scdere semnificativ a parametrilor
reproductivi (Harrison i col., 1990).
n studiul nostru am nregistrat att n cazul L1, ct i n cel al L2, valori ale
insulinei (INS) situate ntre limitele fiziologice. n ambele loturi, valorile INS au
crescut semnificativ (p<0,05) de la momentul parturiiei, la 30 i 45 de zile pp. La
nivel individual, trebuie valorile sczute ale INS nregistrate la vacile cu index de
inseminare inferior, att pentru L1 (7,33U/l), ct i pentru L2 (7,5 U/l). Aceste
valori corespund punctului de vedere expus de Wathes i col. (2003). Astfel, n
puerperiumul timpuriu, cnd vacile se parcurg bilanul energetic negativ,
receptorii pentru somatotropin sunt inactivi, consecina fiind un nivel crescut al
hormonului somatotrop, precum i unul sczut al INS i al IGF1. Chiar dac nu au

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fost evideniate diferene semnificative ntre valorile medii nregistrate la ambele


loturi, indiferent de momentul de referin, parametri reproductivi
necorespunztori (rata gestaiei i indexul de inseminare) ar putea fi determinai
de nivelurile INS.
Slaba rat a concepiei la prima inseminare ar putea fi influenat de
nivelurile INS la 30 i 45 de zile pp, dup cum am observat n acest studiu.
Rezultatele noastre sunt n acord cu cele prezentate de Beam i Butler (1999) i
Butler (2000). Aceti autori afirm c fertilitatea necorespunztoare a vacilor de
lapte este determinat de bilanul energetic negativ nregistrat n puerperium
precum i de nivelurile sczute ale INS i IGF1.
Nivelurile serice ale IGF1 au nregistrat curbe similare celor parcurse de
nivelurile INS, la ambele loturi. Concentraiile IGF1 cresc pe msura derulrii
puerperiumului, dovedind capacitatea ficatului de a rspunde necesitilor
energetice crescnde impuse de producerea laptelui. Chiar dac nivelurile IGF1
sunt mai sczute la L2 comparativ cu L1 la 7 i 15 zile pp., nu exist diferene
semnificative (p>0,05) ntre valorile acestor parametri nici la momentele
menionate i nici la 30 sau 45 de zile pp. Rezultatele noastre sunt contrare celor
prezentate de Snijders i col. (2001), conform crora valorile IGF1 sunt
semnificativ sczute la vacile cu producii mari de lapte comparativ cu cele cu
producii medii, n intervalul cuprins ntre 2 i 10 sptmni post partum.
Trebuie subliniat faptul c nivelurile sczute ale glicemiei, insulinei i IGF1
determin reducerea producerii estrogenilor n foliculul dominant, inhibnd
ovulaia. Este probabil cazul celor dou femele din L1 i al celor trei din L2, care
au avut nevoie de trei sau mai multe inseminri pentru a rmne gestante. Dac n
primele patru sptmni ale puerperiumului nivelul optim al IGF1 este esenial
pentru creterea i dezvoltarea folicular, ulterior este benefic pentru
steroidogenez.
n cazul vacilor cu producie medie de lapte (L1), valoarea medie a BCS la
60 de zile pp a sczut cu 0,38 de uniti comparativ cu momentul parturiiei,
indicnd un efort metabolic echilibrat, suportat corespunztor de ctre organism.
Scderea valorilor acestui parametru a fost de 0,53 uniti pentru L2, ambele
valori fiind inferioare celor prezentate de ali autori. Astfel, Ling i col. (2003) au
raportat o scdere de 1,51 a BCS la acele vaci care au avut antepartum un BCS
mai mare de 3,5 i o scdere de 0,89 n cazul unui BCS antepartum inferior valorii
de 3,5 .
La momentul parturiiei, valorile medii ale BCS au fost de 3,03 (L1) i 3,17
(L2), scderea acestor indici fiind semnificativ n ambele loturi att la 30 ct i la
60 de zile pp, moment la care au atins valori similare (2,65 pentru L1 i 2,64
pentru L2). Femelele care nu au rmas gestante consecutiv primei inseminri au
pierdut ntre momentul parturiiei i 60 de zile pp ntre 0,5 i 0,75 uniti BCS
(L1) i maximum o unitate BCS (cazul unei femele din L2). Avnd n vedere
rezultatele obinute putem afirma c slabele performane reproductive nu sunt

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influenate de variaiile BCS, n special dac ne raportm la rezultatele lui Ling i


col. (2003), conform crora vacile care pierd mai mult de o unitate BCS la debutul
lactaiei nregistreaz o rat a concepiei la prima inseminare cuprins ntre 17 i
38%.
Modificrile BCS reprezint un bun indicator al bilanului energetic (Britt,
1992). Dac bilanul energetic negativ timpuriu influeneaz rata concepiei la
prima inseminare, pierderea BCS ar trebui s fie mai nsemnat la vacile care nu
rmn gestante, comparativ cu cele care rmn, dup cum am observat c s-a
petrecut cu femelele lotului 1, dar nu i cu cele ale lotului 2.
Bilanul energetic negativ este corelat de asemenea cu valori sczute ale
glicemiei i ale IGF1. Harrison i col. (1989) au semnalat faptul c glicemia
sczut se regsete la femelele cu producii mari de lapte i performane
reproductive modeste. Rezultatele noastre, conform crora nivelul glicemiei nu
este corelat cu performanele reproductive se nscriu pe direcia celor prezentate
de Kapel i col. (1984).
Aceste valori, corelate cu nivelul sczut al progesteronului dup prima I.A.
ofer o explicaie obiectiv pentru procentul sczut al fertilitii rezultat
consecutiv primei nsmnri artificiale.
n cazul ambelor loturi (tabelul 3) rata concepiei la prima I.A. (40% pentru
L1 i 30% pentru L2) se situeaz mult sub nivelul acceptabil n cadrul
rentabilitii economice, ns se ncadreaz n tendina actual de descretere a
fertilitii evideniat pe plan mondial (Bousquet i col., 2004).
Tabelul 3
Principalii parametri reproductivi ai celor dou loturi (X Sx)

Prima IA (zile post Rata concepiei Indicele de Service period


Lotul
partum) la prima IA (%) inseminare (zile)
L1 46,6 3,11 40 2 70,221,875
L2 53 3,21 30 2,1 77,120,425

Chiar dac studiul nostru a fost ntreprins pe un numr redus de femele


(L1, n=10 ; L2, n=10), diferena semnificativ dintre valorile ratei concepiei la
prima I.A. evideniaz relaia de invers proporionalitate existent ntre nivelul
produciei de lapte i fertilitate.
Indicele de inseminare se afl la cote mult peste cele acceptate i semnalate
n literatur (Van der Lende, 2002). n studiul menionat, la un service period de
71,7 zile a corespuns indice de inseminare de 1,28, iar la un service period de 84,7
zile, un indice de inseminare de 1,49.
Dei intervalul dintre parturiie i instalarea unei noi gestaii, att pentru
lotul 1, ct i pentru lotul 2 se ncadreaz n limitele acceptate, exist o diferen
semnificativ (p>0,05) ntre valorile medii ale acestui parametru nregistrate
pentru L 1 (70,2 21,875 zile) i L 2 (77,1 20,425 zile).

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CONCLUZII

Nivelurile progesteronului i ale hormonilor tiroidieni nregistrate n jurul


momentului inseminrii sunt decisive pentru soarta produsului de concepie.
Nivelurile sczute ale T3 i T4 dup 45 de zile post partum pot influena negativ
instalarea gestaiei consecutiv primei nsmnri artificiale.
Glicemia crete constant pe msura avansrii n puerperium, valorile medii
nregsitrate fiind mai sczute n cazul vacilor cu producii mari de lapte.
Nivelurile relativ sczute nregistrate n ambele loturi pot fi corelate cu slaba rat
a concepiei la prima inseminare.
Nivelul insulinei crete semnificativ pentru ambele loturi la 30 i 45 de zile
post partum comparativ cu momentul parturiiei. Valorile insulinemiei la 30 i 45
de zile post partum influeneaz rata concepiei.
Vacile cu producie mare de lapte din rasa BNR au valori serice ale IGF1
mai sczute comparativ cu femelele cu producie medie de lapte, diferenele
nefiind ns semnificative. Pe msura scderii valorii IGF1 se constat c sunt
necesare tot mai multe inseminri pentru instalarea gestaiei.
Raporat la momentul parturiiei, scderea BCS a fost semnificativ la 30 i
60 de zile post partum, pentru ambele loturi.
Referitor la momentul apariiei post partum a primului estru precum i la
durata intervalului parturiie-inseminare fecund, exist diferene semnificative
ntre vacile BNR cu producie medie de lapte i cele cu producie mare.
Rezultatele obinute ne determin s afirmm c n cazul vacilor de lapte
din rasa Blat cu Negru Romneasc, indiferent de nivelul produciei de lapte,
performanele reproductive n perioada puerperal sunt influenate de nivelurile
hormonilor tiroidieni, ale glicemiei, insulinei i IGF1, dar nu i de variaiile BCS.

BIBLIOGRAFIE

1. Beam S.W., W.R. Buttler, 1999 - Effects of energy balance on follicular development
and first ovulation in postpartum dairy cows. JRF Suppl. 54:411-424.
2. Buttler W.R., 2000 - Nutritional interactions with reproductive performance in dairy
cattle. Anim. Reprod. Sci. 60-61:449-457.
3. Bousquet D., Bouchard E., DuTremblay D., 2004 - Decreasing Fertility in Dairy
Cows: Myth of Reality? , 23rd World Buiatric Congress 2004, 239-247
4. Britt, J.H., 1992 - Influence of nutrition and weight loss on reproduction and early
embryonic death in cattle, Proceedings of the XVII World Buiatrics Congress, St.
Paul, MN, 2, 143 - 149.
5. Diskin, M.G., Boland, M.P., Dunne, L.D., Sreenan, J.M., 2001 - The effect of
changes in freed intake around the time of insemination on systemic
progesterone and embryo development and survival in cattle. In: Diskin, M. G.,
(Ed.): Fertility in the high-producing dairy cow. British Society of Animal Science,
Edinburgh, Occasional publications, 26, 2, 211-217
6. Ferguson, J.D., David, T., Galligan, T., Thomsen, N., 1994 - Principal description
of body condition score in Holstein cows, J. Dairy Sci, 77, 2695 - 2703.

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7. Harrison, R.O., Ford, S.P. Young, J.W., Conley, A.J., Freeman A.E., 1990 -
Increased milk production versus reproductive and energy status of high
producing dairy cows, Journal of Dairy Science, 73, 2749 - 2758
8. Harrison, R.O., Young, J.W., Freeman, A.E., Ford, S.P., 1989 - Effects of
lactational level on reactivation of ovarian function, and interval from parturition to
first visual oestrus and conception in high producing Holstein cows, Anim. Prod.,
49: 23.
9. Huszenicza, Gy., Kulcsar, M., Nikolic, J.A., Schmidt, J., Korodi, P., Katai, L.,
Dieleman, S., Ribiczei-Szabo, P., Rudas, P., 2001 - Plasma leptin
concentration and its interrelation with some bloode metabolites, metabolic
hormones and the resumption of cyclic ovarian function in postpartum dairy cows
supplemented with Monensin or inert fat in feed. In: Diskin, M. G., (Ed.): Fertility
in the high-producing dairy cow. BritishSociety of Animal Science, Edinburgh,
Occasional publications, 26, 2, 405 - 409.
10. Jooritsma, R., Wensing, Th., Kruip, TAM., Petu, LAM., Noordhuizen, J.P., 2003 -
Metabolic changes in early lactation and impaired reproductive performance in
dairy cows, Vet. Res., 34, 11 - 26.
11. Kapel, L.C., Ingraham, R.H., Morgan, E.B., Zeringue, L., Wilson, D., Babcock,
D.K., Stat, M., 1984 - Relationship between fertility and blood glucose and
cholesterol
12. Ling, K., Jaakson, H., Samarutel, J., Leesmae, A., 2003 - Blood metabolites of
Estonian Holstein cows and their relation to some fertility parameters,
Veterinarija ir Zootechnika. T. 24 , 46, 94 101.
13. Lucy M.C., 2000 - Regulation of ovarian follicular growth by somatotropin and
insulin-like growth factors in cattle. J. Dairy Sci. 83:1635-1647.
14. Lucy M.C., 2001 - Reproductive loss in high-producing dairy cattle where will it
end? J Dairy Sci 84:1277-1293
15. Nikolic, Judith Anna, Ratkovic Marija, Nedic Olgica, 1996 Determination of
insulin-like growth factor I by radioimmunoassay, J. Serb. Chem. Soc. 61, 1149
1157.
16. Opsomer G., Grohn Y.T., Hertl J., Coryn M., Deluyker H., de Kruif A, 2000 - Risk
factors for post partum ovarian dysfunction inhigh producing dairy cows in
Belgium: a field study. Theriogenology 53:841-857.
17. Pavlata L., Prasek J., Filipek J., Pechova A., 2004 - Influence of parenteral
administration of selenium and vitamin E during pregnancy on selected metabolic
parameters and colostrums quality in dairy cows at parturition. Vet. Med. Czech,
49(5):149-155
18. Pechova A., Illek J., Liska I., Halouzka R., Pavlata L., 2000 - Concentration of free
amino acids in blood plasma of dairy cows with developing hepatic steatosis.
Acta Veterinaria Brno 69(2):93-99
19. Pezzi C., Accorsi P.A., Vigo D., Govoni N., Gaiani R., 2003 - 5-deiodinase activity
and circulating thyronines in lactating dairy cows. J Dairy Sci 86:152-158
20. Pusta Dana-Liana, 2004 - Influena radiaiilor calorice solare asupra vacilor de lapte
n condiiile climatice din Transilvania. Universitatea de tiine Agricole i
Medicin Veterinar Cluj-Napoca, Tez de doctorat, pag. 158-169
21. Rabiee, A.R., Leon I.J., Gooden, J.M., Miller, B.G., 1999 - Relationships among
metabolites influencing ovarian function in the dairy cow, Journal of Dairy
Science, 82:39-44.
22. Snijders, S.E.M., Dillom, P.G., OFarrel, K.J., Diskin, M., Wylie A.R.G.,
OCallaghan, D., Rath, M., Boland, M.P., 2001 - Genetic merit for milk
production and reproductive success in dairy cows, Animal Reproduction
Science, 65, 17 31.

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23. Staples C.R., Thatcher W.W., Clark J.H., 1990 - Relationship between ovarian
activity and energy status during the early postpartum period of the high
producing dairy cow. J Dairy sci 73:938-947
24. Van der Lende T., 2002 - Physiological aspects of reproduction and fertility in dairy
cows. J Dairy Sci 85 (6): 1578-1585
25. Wathes D.C., V.J. Taylor, Z. Cheng, G.E. Mann., 2003 - Follicle growth, corpus
luteum function and their effects on embryo development in postpartum dairy
cows, Reproduction Supplement 61:1-19

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INTRAUTERINE INSEMINATION OF SOWS


NSMNAREA ARTIFICIAL INTRAUTERIN LA SCROAF

P. ROCA, L. RUNCEANU,
D. DRUGOCIU, V. PACHIANU
U.S.A.M.V. Iai

For obtaining high and constant fecundity and natality at sows, in


our country are being used doses containing between 3 and 5 billion
spermatozoids. Our research was oriented into a new insemination method,
called intrauterine, which consist in depositing the seminal material in
uterus, thus reducing the number of inseminated spermatozoids. For this
was used the DeepgoldenpigTM device. This method was compared with the
classical artificial insemination, which implies the use of the Goldenpig
device and depositing the seminal material at cervical level. Doses
containing 3, 2, and 1 billion spermatozoids, diluted with X Cell gel were
packaged in 80 ml Cochette bags. The females with a weaning to estrus
interval of 2 11 days were inseminated twice at an interval of 12 hours.
Pregnancy was determined at 28 30 days from insemination using an
ultrasound scan. The standard inseminating method produced fecundity
rates of 90.6%, 87.9%, and 60.5% per doses with 3, 2, and 1 billion
spermatozoids while the intrauterine method obtained rates of 91.6%,
90.8% and 59.5%. In similar way the farrowing rates was 90.2%, 87.4%,
and 59.5% in case of classical method and 90.9%, 90.3%, and 81.1% for
intrauterine method. Significant differences were obtained using 1 billion
spermatozoids doses. In conclusion intrauterine insemination is simple, safe
and effective, and allows the sperm dose to be reduced to 1 billion
spermatozoa.

Key words: swine, gestation, fertility, prolificacy.

Practiced for many years, artificial insemination of pigs implies the use of 3
5 billions spermatozoa per dose for obtaining a high fecundity. Seminal material
can be crude, freshly diluted, diluted and conserved for 3 7 days at 17oC or
frozen (1, 3, 4).
For insemination can be used various kinds of catheters, some of them who
simulates the corkscrew tie of boars penis but all of them allow seminal material
to be deposited at cervical level. For overcome the barrier represented by the
cervix in the way of spermatozoids, is necessary to deposit a large amount of
seminal material at cervical level to ensure a sufficient number of spermatozoa in
the isthmus of the oviduct. Overcoming this barrier and depositing the seminal
material directly in uterus allows the reducing of the number of spermatozoids per
dose, assuring a sufficient number of spermatozoids at the place of fecundity (2,
5).

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Knowing these aspects, the scope of this study was the comparative testing
of the two methods of insemination, intrauterine and classical, with the reducing
of the number of spermatozoids per dose.

MATERIALS AND METHOD

The study was performed on 2685 PIC Camborough sows with a weaning
to estrus period of 2 11 days. The seminal material used was diluted with X
Cell packaged in 80 ml Cochette bags. The sows were reinseminated in 12 hours.
The intrauterine method was tested comparative with the classical method
of insemination. In the classical method is being used the Goldenpig standard
device, which allows seminal material to be deposited at cervical level. The
intrauterine method implies the use of the DeepgoldenpigTM inseminating device,
who is in fact a DeepgoldenpigTM standard catheter, through which is inserted a
narrow inner tube with a diameter of 4 mm and which extends 200 mm beyond
the tip of the outer catheter, reaching the body of the uterus, where the seminal
material is deposited. In parallel with the testing of the two methods, was reduced
the number of spermatozoids per dose, sows being inseminated with doses of 3,
2, and 1 billion spermatozoids in both methods. Gestation was established with
ultrasounds at 28 30 days from insemination.

RESULTS AND DISCUSSIONS

The comparative results for the two methods and the three concentrations
are presented in Table 1.
Tabel 1
Comparison between fecundities and fertilities obtained after the
Inseminations with the standard device Goldenpig and the newer device
Deepgoldenpig

Dose Inseminated Gestation Parturition Litter size


Device
(billions) No. % No. % No. % No. %
3 450 408 90.6 406 90.2 5034 12.3
Goldenpig 2 415 365 87.9 363 87.4 4428 12.1
1 200 123 60.5 119 59.5 1167 9.8
3 650 596 91.6 591 90.9 7388 12.5
DeepgoldenpigTM 2 620 563 90.8 560 90.3 6886 12.2
1 350 292 83.4 284 81.1 3378 11.8
Total 2685 2347 2323 28281

The rates of pregnancy in case of using doses with 3 billions spermatozoids


were, with the classical method, 90.6% and 91.6 with the intrauterine method and
they did not differ significantly from the 2 billion spermatozoids doses. This
demonstrates that, for a good fertility, are necessary at least 2 billions
spermatozoids per dose. For doses of 1 billion spermatozoids, inseminated with
the classical method, the fertility rates are only of 60.5%, which means that this

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concentration is insufficient to assure the necessary amount of functional


spermatozoids in utero-tubal junction and implicit in the isthmus of the oviduct.
With intrauterine insemination of doses with 1 billion spermatozoids were
obtained a higher fecundity rate, 83.4%. The obtained results demonstrates that
the intrauterine insemination with a reduced number of spermatozoids is not an
disadvantage.
Analyzing the parturition, with the classical method of insemination, the
rates are 90.2%, 87.4% and 59.5% for doses of 3, 2 and 1 billion spermatozoids,
while, for the same doses, with the intrauterine method the rates of the parturition
are 90.9%, 90.3% and 81.1%. Only in case of insemination of 1 billion
spermatozoids with the classical method the differences are significant from the
doses of 2 and 3 billions spermatozoids inseminated with the same method.
A similar evolution had and the litter size. With the classical method the
litter size was 12.3, 12.1 and 9.8, while with the intrauterine method the litter size
was 12.5, 12.2 and 11.8.
Considering the results can affirm that intrauterine insemination at sows is
simple, safe and effective, allowing reducing the number of spermatozoids per
dose at 1 billion.

CONCLUSIONS

With the classical insemination method the fecundity rates were 90.6%,
87.8% and 60.5%, for doses with concentrations of 3, 2 and 1 billion
spermatozoids.
With the intrauterine insemination method the fecundity rates were 91.6%,
90.8% and 83.4%, for doses with the same concentrations.
Similar results were obtained for the farrowing and the litter size.
Only with the use of 1 billion spermatozoids doses the differences were
significant in favor of the intrauterine method.
The use of the intrauterine insemination method, associated with the
reducing of the concentration of the doses, offer important earnings by the more
rational usage of the genetically superior boars.

BIBLIOGRAPHY

1. Bogdan, A.T., Mantea, t., Bogdan Dorina, 1999 Tratat de reproducie i


nsmnri artificiale la suine. Ed. Tehnic Agricol, Bucureti.
2. Krueger, C., Rath, D., 2000 Intrauterine insemination in sows with reduced sperm
number. Reprod. Fertil. Dev. 12 : 113-7.
3. Runceanu, L., Cotea, C., 2001 Reproducie, obstetric i ginecologie veterinar.
Ed. Ion Ionescu de la Brad Iai.
4. Runceanu, L., Drugociu, D., Roca, P., Anton, C., 2002 Reproducie, obstetric
i andrologie clinic. Casa de editur Venus, Iai.
5. Watson, P.F., Behan, J.R., 2002 Intrauterine insemination of sows with reduced
sperm number: results of a commercially based field trial. Theriogenology. 57 :
1683-1693.

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CERCETRI ASUPRA UNOR CONSTANTE SANGUINE


N ENDOMETRITELE SCROAFELOR
RESEARCHES ON SOME SANGUINE CONSTANTS IN SOWS ENDOMETHRITIS

L. RUNCEANU, D. DRUGOCIU, P. ROCA


U.S.A.M.V. Iai

Researches made shows that approximative 25.84% from females


presents modified puerperal remaining with mucopurulent aspect, in bigger
rate at multipares .Females with genital infections presents a significant
decrease of erythrocytes average values in comparison with witness lot
(5,93 0,92 witness lot in comparison with 4,91 0,15 experimental lot
phenomenon meet and in the case of hemoglobin. Leucocytes number shows
a growth at the lot with endomethritis in comparison with the witness lot
(12,5 2,46 of 11,36 2,92)

INTRODUCERE

Perioada puerperal la scroaf dureaz 2-8 zile, loiile eliminndu-se


complet n 2-5 zile (Bogdan, A.T., i col., 1999). Persistena loiilor peste aceast
limit de timp, constituie o stare patologic, atestnd o stare de infecie
consecutiv traumatizrilor tractusului genital n perioada parturial sau a
reteniilor placentare pariale i contaminrilor ascendente (Berner, 1980, Mandiu
i col., 1980, Boitor, 1981, Runceanu Margareta, 1992, Runceanu, L., Cotea, C.,
2001, Broiu, A., Seiciu, Fl., 2004, Thibault, 1991, Tarocco, 1986, Vaissaire, J.P.,
1977, Bilrot, P., i col., 1980, Drugociu, D., 2005).
Cu toate c boala poate evolua uneori fr apariia unor semne generale
evidente, se produc unele modificri decelabile numai prin examenele paraclinice.

MATERIAL I METOD DE LUCRU

Cercetrile au fost efectuate pe un numr de 35 scroafe ce prezentau


endometrit puerperal. Femelele au fost n vrst ntre 1,5 i 3 ani iar condiiile
de ntreinere au fost corespunztoare.
Probele de snge au fost recoltate n primele zile post partum, cnd a fost
diagnosticat afeciunea. Prelevarea s-a efectuat prin puncia confluentului
jugular iar ca anticoagulant a fost folosit EDTA.
Examinarea seriei roii i a numrului de leucocite s-a realizat folosind
tehnicile uzuale. Cantitatea de hemoglobin a fost determinat prin metoda Sahli.
Lotul martor a fost constituit din 30 scroafe n aceiai faz puerperal, dar
la care nu a fost diagnosticat endometrita.
Rezultatele cercetrilor au fost prelucrate statistic: media eroarea standard
a mediei i semnificaia.

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REZULTATELE I DISCUII

Analiznd variaia constantelor hematologice luate n studiu constatm


pentru unele din ele, valori ce pun n eviden existena fenomenului de boal cu
repercusiuni asupra strii generale a organismului.
Tabelul 1
Valorile medii ale constantelor hematologice

Nr. Scroafe cu
Denumirea constantei Lot martor
Crt. endometrit
1 Globule roii mil/mm3 5,93+0,92 4,91+0,15
2 Hemaglobulina mil/mm3 14,4+0,93 10,02+0,19
3 Leucocite mil/mm3 11,36+2,92 12,5+2,46
4 Euzinofile % 3,5+0,22 3,82+0,14
5 Neutrofile % 44,1+1,40 49,12+0,96
6 Bazofile % 0,6+0,1 0,7+0,04
7 Limfocite % 36+1,37 40,97+1,05
8 Monocite % 5,4+0,26 4,99+0,21

Din datele prezentate constatm c la scroafele cu endometrit valorile


medii ale eritrocitelor (4,91+0,15) au sczut foarte semnificativ n comparaie cu
valorile medii ale lotului martor (5,92+0,22).
n mod asemntor scade i cantitatea de hemoglobin : de la 0,02+0,19 la
scroafele cu endometrit, la 14,4+0,93 la lotul martor. Modificrile aprute n
evoluia elementelor seriei roii, se coreleaz cu starea particular a animalelor n
perioada post partum, la care se adaug procesul patologic.
Cercetrile referitoare la numrul total de leucocite i la elementele
formulei leucocitare atest urmtoarele aspecte:
- la femelele cu endometrit valoarea medie a lotului este de 12,5+2,46,
mai crescut, dar nesemnificativ fa de valoarea medie a lotului
martor (11,36+2,92).
- evoluia elementelor albe n formula leucocitar pledeaz pentru
existena unui proces inflamator, o reacie de sensibilizare a
organismului. Astfel, cresc semnificativ neutrofilele (49,12+8,96) la
femelele bolnave, fa de (44,1+1,40) la lotul martor, limfocitele
(40,97+1,05) la lotul cu endometrit fa de (36+1,37) la lotul martor
i eozinofilele (3,82+0,14) la animalele bolnave fa de (3,5+0,22) la
lotul martor.
Creterea neutrofilelor atest inflamaiile septice de la nivelul organelor
genitale, dup cum creterea semnificativ a limfocitelor poate fi asociat cu
natura infecioas a procesului inflamator conferind astfel organismului
capacitatea de a lupta mpotriva agentului patogen.

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Monocitele au avut o valoare de 5,4+0,26 la lotul martor fa de 4,99+0,21,


iar bazofilele o cretere nesemnificativ la lotul experimental (0,7+0,04) fa de
lotul martor (+0,1).

CONCLUZII

1. Valorile medii ale numrului de eritrocite la femelele cu endometrit


scad semnificativ n comparaie cu lotul martor (4,91+0,15 fa de 5,92+0,22).
2. Cantitatea de hemoglobin prezint o scdere la femelele cu endometrit
(10,02+0,19 fa de lotul martor).
3. La femelele cu endometrit, valoarea medie a numrului total de
leucocite este de 12,5+2,46, mai crescut dect la lotul martor.
4. Evoluia elementelor albe n formula leucocitar pledeaz pentru un
proces inflamator, crescnd semnificativ neutrofilele, limfocitele i eozinofilele.

BIBLIOGRAFIE

1. Berner, K., 1980 Reasons for culling of gilts in relation to age at first mating. The
Vet. Bul. 50.
2. Bilrot, P., Dekeyser, P., 1980 Rol of infections in the SMEDI and abortion
syndrome on Sows. The Vet. Bul. 50.
3. Boitor, I., 1985 Fiziologia i fiziopatologia reproduciei animalelor de ferm. Ed.
Ceres, Bucureti.
4. Broiu, A., Seiciu, Fl., 2004 Tratat de reproducie la animale. Ed. ALL, Bucureti
5. Bogdan, A.., i col., 1999 Tratat de reproducie i I.A. la suine. Ed. Cerces,
Bucureti.
6. Drugociu, D., 2005 Bolile obstetrical ginecologice la animale. Ed. Ion Ionescu de
la Brad, Iai.
7. Runceanu Margareta, 1992 Cercetri privind influena afeciunilor genitale asupra
fertilitii suinelor crescute n sistem industrial. Tez de doctorat.
8. Runceanu, L., Cotea, C., 2001 Reproducie, obstetric i ginecologie veterinar.
Ed. Ion Ionescu de la Brad, Iai.
9. Tarrocco, C., 1986 Tulburri ale aparatului de reproducie femel produse de cauze
neinfecioase (la suine). Med. Vet. XII, Bucureti.
10. Thibault, C., Levasseur, M.C., 1991 La reproduction chez les mammiferes et
lhome. Ed. Marketing. Paris.
11. Vaissaire, J.P., 1972 Sexualite et reproduction des mammiferes domestiques et de
laboratoire. Ed. Maloine, Paris.

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THE THERAPY OF PLACENTA RETENTION WITH PGF2


TERAPIA RETENIEI PLACENTARE CU PGF2

M.I. VIZMAN

The parturition mechanism is very complex and it consist, generally,


of passing the uterus muscle from a relative stillness state to an intense
contraction state, but only after the cervix is opened.
The parturition mechanism has at his base two theories: a
neuroendocrine theory and an imunologic theory. According to the first
theory the neurohormonal complex represented by the cortical centres,
throughout the hipotalamus-pituitary-uterine axe and the hipotalamus-
pituitary-corticosuprarenalian foetal, axe conditions the parturition.
The gestagen, estrogen and prostaglandins hormone levels, and
specially the hormons from PGF2 influence the parturition. According to
these neurohormonal theory, the beginning and the maintenance of the
foetal and the foetal annexes expulsion is accomplished as a consequent of
the disruption of the balance between the gestagen and estrogen hormones.

In cow the parturition is harder than in the rest of the farm animals because
of the bone structure in the pelvic aria that is unfavourable to birth doe to the next
particularises:
- the anterior opening is lining to vertical axe and is hardening the foetal
engage in the birth canal
- the subcotiledonian crest are very well developed
- the insertion of the prepubian ligament is under the anterior margin of
the ischiopubian simphise, fact that is making the pelvic axe to look
like a broken line.
- the ischiadic arcade is narrow and the ischiadic tuberoses have three
ends and are high is the last part of the pelvic aria
The parturition is easier in the sternal-abdominal or coast-abdominal
position. The uterine involution is starting right after the closer of the parturition,
and at 10-15 minutes

MATERIAL AND METHOD

The clinical studies were conducted on 20 cows at which after the


parturition there was the retention of the foetal annexe. These cows were treated
according to the next protocol: at 24 hours from parturition there was
administrated 500 mg of cloprostenol (synthetic analogue of PGF2), 500 ml from
a suspension containing saline solution, oxitetraciline 5g and 30 ml of ethylic
alcohol 70 volume, was administrated inside the uterus. In some cases the
treatment was repeated.

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This treatment was applied to the females that suffer from foetal annexes
retention and which represented the experimental lot.
The objectives that we followed were:
- the time interval from parturition to the elimination of the foetal annexe
- the vaginal secretions dynamics
- the time interval in which the uterine involution was completed
(monitored throw trans-rectal exam)
- the time interval from the resume of the sexual activity
The cows that were threaded with prophylactic treatment represented the
witness lot with SEL-E-VIT, used to reduce the incidence of foetal annexe
retention
The mean duration of pregnancy at the studied cows was 285 days. After
the foetal annexe retention there was no alteration of the general being of the
cows, there was no febrile state registered.

RESULTS AND DISCUSSION

The results regarding the elimination of the foetal annex after the
parturition, obtained after the clinic study conducted at cows with placenta
retention after the PGF2 treatment are presented in the table 1
Tabel 1
Results attained

Parturition-
No. Registration Fecund Artificial Parturition Treatment fetal annexe
crt. no. Insemintion Data data data elimination
interval
1. 01008 16.02.2003 10.12.2003 11.12.2003 9 days
2. 010082 10.02.2003 11.12.2003 12.12.2003 9days
3. 010076 02.03.2003 21.12.2003 22.12.2003 9 days
4. 010084 25.03.2003 12.01.2004 13.01.2004 7 days
5. 010092 02.04.2003 21.01.2004 22.01.2004 9 days
6. 010016 04.04.2003 25.01.2004 26.01.2004 9 days
7. 010030 21.04.2003 05.02.2004 06.02.2004 8 days
8. 010086 15.05.2003 20.02.2004 21.02.2004 8 days
9. 743 17.05.2003 27.02.2004 28.02.2004 9 days
10. 640 25.05.2003 02.03.2004 03.03.2004 7 days
11. 652 15.06.2003 07.03.2004 08.03.2004 8 days
12. 749 28.06.2003 20.03.2004 21.03.2004 8 days
13. 546 10.07.2003 03.04.2004 04.04.2004 9 days
14. 010038 21.07.2003 12.04.2004 13.04.2004 9 days
15. 010046 23.07.2003 21.04.2004 22.04.2004 8 days
16. 010062 27.07.2003 28.04.2004 29.04.2004 7 days
17. 757 29.05.2003 27.03.2004 28.03.2004 9 days
18. 610 15.05.2003 26.03.2004 27.03.2004 8 days
19. 010070 01.07.2003 13.04.2004 14.04.2004 7 days
20. 010048 29.06.2003 08.04.2004 09.04.2004 7 days

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The percent representation of the dynamic of the foetal annex elimination is


represented in the table 2.
Tabel 2
The dynamics of the foetal annexes elimination

Parturition-fetal annexe elimination interval


No. Cows 7 days 8 days 9 days
Nr. % Nr. % Nr. %
20 5 25 6 30 9 45

From the table we can see that at the majority of the cows the placenta
elimination was accruing at 9 days after the parturition. The percent of
elimination in the 7,8 and 9 day after the parturition is presented in the table 2.
The second objective that was monitored after the administration of the
treatment is the dynamic of the genital secretion.
The genital secretions in the first 10 days are more abundant at the treated
cows, they are reddish in colour. The clearing and the reduction of the volume of
the genital secretions started in the 12-day after the parturition.
The genital secretions was clear in the 16-day, and at 21 days after the
parturition the vaginal secretions stopped.
In regarding to the uterine dynamic, there was observed a rapid involution
of the uterus immediately after the elimination of the conception product. The
uterus is at 60% from the volume before the parturition.
From this moment until the elimination of the foetal annexe the uterine
involution wasnt noticed or it was much reduced, undetectable at the transrectal
exam. From the moment of the foetal annexe elimination, the involution was
resumed, and in the 15-16 days form the parturition the uterus was at 50% volume
from the volume at parturition. The uterine involution was completed at 22 days
after the parturition.
In respect to the resuming of the ovarian activity, the results were varied.
The following of the ovarian activity was made at 100 days after the parturition.
The period until the resuming of the oestrus cycling was extended in the majority
of the cases; this fact has as consequence the extending of the service period
duration and at the calving interval. The presenting of the resuming of the ovarian
activity after parturition is presented in the table 3.
Table 3
The resume of the ovarian activity

Nr.crt. Nr. matricol Nr. de zile scurse pn la reluarea ciclului estral


1. 01008 Peste 100
2. 010082 96
3. 010076 82
4. 010084 70
5. 010092 64

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Nr.crt. Nr. matricol Nr. de zile scurse pn la reluarea ciclului estral


6. 010016 38
7. 010030 68
8. 010086 85
9. 743 48
10. 640 Peste 100
11. 652 68
12. 749 64
13. 546 48
14. 010038 43
15. 010046 50
16. 010062 82
17. 757 Peste 100
18. 652 75
19. 010070 56
20. 00148 52

CONCLUSIONS

After clinical study of the efficiency of the foetal annexe retaining


treatment after the protocol described by Robertson, the following conclusions
can be made:
- the foetal annexes were eliminated in average 8,33 days
- the clearing of the genital secretions is starting in the 12 day
postpartum, and they become clear in the 16 day and they stop in the
21 day after parturition
- the uterine involution was completed in the 22 day after the parturition
- the resuming of the ovarian activity was between the 38 and 100 days
- after the treatment the foetal annexes were eliminated with out
complications, although the elimination was completed in 9 days
- also it can be noticed that the period of time when the resuming of the
ovary activity took place was extended (even over 100 days), fact that
led to the prolonging of the service period.
- because of these drawback the protocol of treatment throw
Robertsons method is not applicable in the case of the milking cows,
at which the milking period must be 305 days and at which the goal is
to attain one calf from one cow every year
- also the long remanence of the placenta in the uterus can determine
the apparition of endometrites which could lead to the dropping of the
productive potential of the females.

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BIBLIOGRAPHY

1. Ardelean, V., 2003 - Fiziologia si patologia gestaiei si parturiiei la animale. Ed.


Mirton Timioara
2. Broiu, A., Robertson, Ed., Seiciu, Fl., Negoia, D., Petcu, M., Ghencioiu, C.,
2000 - Studii privind o noua terapie a reteniei nvelitorilor fetale la vac, Rev.
Rom. Med. Vet.., vol. XIII, nr. 2, pag. 60-64. Bucureti
3. Bonca, Gh., Cernescu, H., Cristescu, E., Ardelean, V., 1989 - Cercetari privind
profilaxia reteniei anexelor fetale la vac. Sem. Actualiti in patologia animalelor
domestice, vol. XIII, Cluj-Napoca.
4. Cernescu, H., Bonca, Gh., Mircu, C., Ardelean, V., Farca, I., Mari, M., 1992 -
Cercetri privind combaterea endometritelor la vaci cu Germisan si fluide
magnetice. Rev. Rom. Med. Vet., vol.II., Bucureti.
5. Cernescu, H., 2004 - Ginecologie veterinara, Cecma Partner, Timioara.

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ARMONIZAREA CU LEGISLAIA EUROPEAN


SOLUIA COMPETITIVITAII PE PIAA EUROPEAN

ASOCIAIA OAMENILOR DE AFACERI IAI 2003

Contientiznd importana politicii agricole comune, Asociaia Oamenilor


de Afaceri (AOA) Iai 2003, n parteneriat cu Direcia Agricol i Dezvoltare
Rural Iai, a lansat proiectul AVI-RO n EUROPENE, proiect finanat de
UNIUNEA EUROPEAN prin programul Phare 2003 Consolidarea Societii
Civile n Romnia.
Obiectivul general al acestui proiect este accelerarea procesului de
armonizare i implementare a acquis-ului comunitar n domeniul agriculturii, ca
ramur integrant a politicii agricole comune.
Obiectivele specifice constau n:
1. nfiinarea unui departament de afaceri europene n cadrul AOA Iai 2003
care s asiste firmele din domeniul agriculturii si industriei alimentare din Iasi i pe
cele din aria creterii psrilor i procesrii crnii de pasre din ara, n demersul lor
de armonizare i implementare a acquis-ului comunitar.
2. Creterea gradului de contientizare a importanei armonizrii cu legislaia
european i implementarea ct mai rapid a acquis-ului comunitar nc din
perioada de pre-aderare a Romniei la UE.
3. Realizarea unui studiu pilot la una din unitile importante din domeniul
avicol.
4. Valorificarea rezultatului studiului pilot prin elaborarea unei metodologii
de armonizare i implementare a acquis-ului comunitar
n cadrul Departamentului Afaceri Europene, realizm documentare i
cercetare n vederea acumulrii de informaii referitoare la acquis-ului comunitar,
norme i standarde europene, acestea centralizndu-se ntr-o baz de date
disponibil gratuit grupului int al proiectului
De asemenea, desfaurm o campanie de contientizare a importanei
implementrii legislaiei europene, distribuind materiale informative (brouri,
pliante, CD-uri) celor din grupul int. Broura InfoACQUIS Informaii
privind adoptarea i implementarea acquis-ului comunitar este disponibil n
format electronic, pe pagina web a proiectului, http://www.aoai.ro/aviro/, i poate
fi procurat n format tiprit, gratuit, la sediul asociaiei.
Prin acest proiect se va edita i un manual metodologic cu algoritmul de
urmat n demersul implementrii acquis-ului comunitar, avnd la baz un studiu
pilot ce se va efectua la o societate comercial din Iai, pentru diagnosticarea
situaiei actuale a acesteia i trasarea msurilor corective necesare implementarii
acquis-ului comunitar, de-a lungul fluxului tehnologic.

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Manualul se va distribui gratuit, ntr-un tiraj de 150 de exemplare i un


numar de 500 de CD-uri cu varianta electronic a studiului, fiind un exemplu de
cum trebuie sa functioneze o firma performant la nivel comunitar.
Mai adaugm c popularizarea rezultatelor finale ale acestui proiect se va
face i prin organizarea unui seminar de 2 zile (7-8 septembrie 2006) la Iai, care
va avea ca participani reprezentani ai grupului int i ca invitai speciali,
consultani i specialiti din domeniul aviculturii.

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OPORTUNITATEA IMPLEMENTRII
MANAGEMENTULUI SIGURANEI ALIMENTELOR
N NTREPRINDERI IEENE
APPROPRIATENESS OF FOOD SAFETY MANAGEMENT IMPLEMENTATION
IN IASI ENTERPRISES

Gh. BRDAN
U.S.A.M.V. Iasi

This study was carried out to evaluate the opportunity and the
necessity to implementation a management system for guarantied the food
safety in two enterprises of Iasi, Romania. The risks for food chain at
various stages of processing were assessed and the reduction of the risk
analyzed. The paper presents the advantages of controlled food dangers for
internal and external medium of alimentary enterprise.

Key words: European laws, food safety, management system.

INTRODUCERE

Stilul de via contemporan este mult diferit fa de cel al generaiilor din


trecut. n ultimii 15 ani, societatea romneasc s-a transformat profund, fiind
influenat de trsturile epocii post-industrializrii i a informatizrii, ce fac parte
din fenomenul complex de globalizare (5). Pe plan social se remarc o schimbare
a conceptului de familie, considerat nc unitatea de baz a societii. Asistm la
o cretere a numrului de familii monoparentale, de persoane celibatare i de
femei care sunt angajate n cmpul muncii. Lipsa de timp i modificarea relaiilor
dintre membrii familiei au condus la apariia de noi obiceiuri culinar. Domeniul
agro-alimentar s-a adaptat noilor cerine, pentru a rspunde schimbrilor profunde
ale nevoilor consumatorilor n prepararea alimentelor (3).
Un rezultat pozitiv al acestor schimbri a impus apariia de noi tehnologii
prin care industria agro-alimentar s poat asigura cantitile necesare de
alimente. n ciuda a numeroase progrese, riscul sanitar legat de alimente nu a
disprut i se propag fie n mod natural, fie accidental att prin contaminare ct
i prin practici greite de fabricare (4). Calitatea i sigurana alimentelor depinde
de eforturile depuse de fiecare actor al lanului alimentar, de la agricultor la
consumator. Sigurana sanitar a alimentelor este afacerea tuturor, de la ferm la
farfurie (6).
Crizele alimentare succesive ale anilor 1990 au alarmat consumatorii i au
incitat puterile publice s pun n practic noi dispozitive de control a siguranei
alimentelor. n majoritatea rilor europene au fost create structuri adecvate

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responsabile de analiza riscurilor i de gestiunea lor. Politica de siguran a


alimentelor este bine ncadrat de norme comunitare i internaionale (2, 5, 7).
Punerea n practic a prevederilor legale referitoare la calitatea i sigurana
sanitar pe ntregul lan alimentar necesit proceduri bine puse la punct i aplicate
n mod permanent, pentru a garanta c alimentul destinat consumatorului este
lipsit de factori care s i pun n pericol sntatea (1).
Obiectivul acestui studiu este de a analiza condiiile de punere n practic a
viitoarelor prevederi legale din domeniul siguranei sanitare a alimentelor, n
uniti agro-alimentare din Iai. Pornind de la cerinele legale i de la situaia
existent, la cteva luni nainte de aderarea Romniei la Uniunea European, se
discut necesitatea implementrii unui sistem de management, eficient, care s
garanteze consumatorului sigurana sanitar a alimentelor oferite.

MATERIAL SI METODA

Studiul a fost efectuat n perioada februarie - mai 2006. Prima etap a


constat din identificarea aspectelor legislative, generale, referitoare la garantarea
siguranei sanitare a alimentelor, aplicate n ara noastr, i mai ales a noilor
hotrri de guvern (2) care transpun n dreptul naional regulamentele europene
din pachetul igien (7). Acestea vor deveni obligatorii ncepnd cu 1 octombrie
2006. Analiza minuioas a prevederilor textelor legislative a fost urmat de
parcurgerea informaiilor prezente pe web-siturile instituiilor naionale i europene
(1, 2, 5). n etapele preliminare de implementare a sistemului HACCP n dou
ntreprinderi din municipiul Iai, un abator i o brutrie, care au ca obiect de
activitate producerea i comercializarea de produse alimentare, s-a efectuat o
analiz a situaiei existente att din punct de vedere a infrastructurii existente ct
i a punerii n practic de proceduri operaionale, n care implicarea personalului
s fie elementul central. Prin coroborarea prevederilor legislative cu posibilitile
de modernizare a ntreprinderilor respective, s-a evideniat importana
implementrii sistemului de management al siguranei alimentului, modalitate de
continuare cu succes a activitii economice.

REZULTATE I DISCUII

Cadrul legal european. Ameliorarea sntii i a ncrederii cetenilor


reprezint elemente foarte importante ale politicii Uniunii europene. Consumatorii
au dreptul de a dori ca alimentele pe care le consum s fie lipsite de pericole
pentru sntatea lor i proprii consumului (6). Sigurana alimentar este un
demers raional efectuat pentru a produce alimente lipsite de riscuri biologice,
chimice sau fizice (4).
Reguli generale n materie de igien a produselor alimentare au fost stabilite
nc din 1993. Experiena a artat c aceste principii au constituit o baz solid
pentru a ine sub control sigurana sanitar a alimentelor. n cadrul politicii
agricole comune, au fost adoptate numeroase alte directive i regulamente pentru

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a stabili obligaii sanitare specifice pentru producerea i punerea pe pia a


produselor alimentare. Ele au redus obstacolele comerului cu astfel de produse
ceea ce a contribuit la realizarea unei piee interioare, asigurndu-se n acelai
timp un nivel ridicat de protecie a sntii publice (6).
Totui, experiena crizelor alimentare din ultimii douzeci de ani a artat c
lanul alimentar trebuie luat n globalitatea sa dac se dorete cu adevrat s se
garanteze sigurana alimentelor.
n prezent politica de siguran a alimentelor din Uniunea european are ca
element distinctiv responsabilitatea productorilor i furnizorilor de a menine
calitatea alimentelor, regulamentele europene fiind cele mai stricte din lume (5).
Abordarea integrat n materie de siguran alimentar vizeaz msuri
coerente pe ntreg lanul alimentar i o supraveghere adecvat, care s garanteze
funcionarea eficace a pieei interioare. Punerea n practic a acestui concept
implic punerea n practic de dispoziii legislative care s asigure controlul
eficace i s evalueze respectul normelor comunitare, s gestioneze relaiile
internaionale cu rile tere i organizaiile internaionale.
Legislaia european n materie de siguran a alimentelor a fost complet
rescris ncepnd cu anul 2000, cnd a aprut Cartea Alb ce prevedea revizuirea
tuturor legilor existente, pentru a le face mult mai transparente i cldite pe o baz
tiinific (6). Primul fruct ale noii viziuni a fost Regulamentul 178/2002, cel care
a introdus noi noiuni ca: analiza riscurilor, principiul precauiei i protecia
consumatorului n materie de siguran alimentar. n plus s-a stabilit nfiinarea
unui organism independent care s lucreze n colaborare cu numeroasele agenii
tiinifice i instituii ale statelor membre. Aceast nou creat Agenie European
pentru Sigurana lanului alimentar (EFSA) furnizeaz date tiinifice asupra
domeniile ce au o relaie direct sau indirect cu sigurana alimentelor, de la
producia primar la livrarea ctre consumator. EFSA se ocup de asemenea cu
evaluarea riscurilor din interiorul lanurilor alimentare i evaluarea tiinific a
tuturor materiilor n raport cu alimentaia (1).
Alt treilea pas important al refacerii legislative a fost publicarea n Jurnalul
Oficial al UE, n data de 30 aprilie 2004, a unui ansamblu de texte oficiale,
regulamentele 852, 853, 854 i 882, numite i Pachetul igien (7). Astfel, au
fost regrupate obligaiile exploatanilor sectorului alimentar n materie de igien a
alimentelor i apoi s regrupate obligaiile n materie de controale oficiale pe care
Statele membre trebuie s le respecte.
Toate aceste noi reguli generale i specifice n materie de igien au ca
principal obiectiv s asigure consumatorului un nivel de protecie ridicat n
materie de siguran sanitar a alimentelor. Fiecare exploatant al sectorului
alimentar, inclusiv cel situat n Iai, la extremitatea estic a UE, trebuie s
supravegheze s nu fie compromis sigurana alimentelor, posibil de realizat doar
printr-o abordare integrat.

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Avantaje ale garantrii siguranei alimentelor. n spiritul


consumatorilor, calitatea i sigurana sunt noiuni primordiale pentru produsele
alimentare. Sigurana alimentelor reprezint conceptul esenial al existenei
unitilor productoare, transformatoare sau distribuitoare de produse alimentare.
Dei vehiculat doar de civa ani, sintagma de sigurana alimentar este prezent
n vocabularul tuturor productorilor dar i al consumatorilor. n contextul
economiei de pia care a devenit funcional n ara noastr, productorii de
alimente din Iai au neles deja c singura cale spre succesul economic, este de a
garanta consumatorului livrarea de produse lipsite de orice pericol ce ar putea
prejudicia sntatea.
Pentru societile analizate s-a constatat c exist dou grupe de factori care
impun introducerea unui sistem de management al siguranei sanitare a
alimentului.
a) Mediul extern al organizaiei este reprezentant de constrngerile legale,
de satisfacerea nevoilor clienilor i a altor organisme. Intrarea n Uniunea
european deschide calea unei confruntri de tip concurenial care presupune
exigene la care trebuie s se adapteze productorii. Pentru a-i putea pune pe
pia produsele, ntreprinderile trebuie s demonstreze c respect cerinele
impuse de legi, directive i regulamente referitoare la produsele respective.
Satisfacia clientului este esenial pentru supravieuirea economic a
ntreprinderilor. Dup convingerea autoritilor c sunt ndeplinite constrngerile
legale, trebuie convini clienii. Clientul este cel care alege produsul, asigurnd
vnzrile. Client este numele de cod pentru unde sunt banii. Acetia vnd
bani cu care cumpr satisfacie; mereu mai mult satisfacie. Nu a satisfacerii
cerinelor sale ci a satisfaciei. Conceptul de management al calitii este centrat
pe creterea continu a satisfaciei clienilor iar managementul siguranei
alimentare permite garantarea continu a fabricrii de produse care s nu duneze
sntii. Prin urmare, n pia, pe terenul pragmaticilor, al oamenilor ordonai,
orientai spre succes, nu este loc pentru vechile metode de organizare.
ntreprinderile analizate depind i de alte organisme care sunt foarte
interesate de viabilitatea afacerilor, mai ales de bncile de credit ce supravegheaz
ndeaproape att evoluia financiar ct i noile constrngeri. Persoanele situate n
imediata vecintate a ntreprinderilor dar i comunitatea local, cea din care fac
parte clienii direci sunt sensibili la variaiile de credibilitate ale organizaiilor.
b) Angajaii ntreprinderilor analizate nzuiesc de asemenea spre
conceptul de siguran alimentar. Nu doar prevenirea amenzilor, a pedepselor, a
pierderilor i motiveaz pe angajai ci mai ales creterea mulumirii sufleteti
generate de lucrul organizat, n care managementul siguranei alimentare este
proiectat, implementat, verificat i actualizat periodic. Pentru a deveni un mediu
compatibil cu stimulii din exterior, principiile legale ale siguranei alimentelor au
fost tratate cu respect, att n cadrul relaiilor verticale ct i orizontale din
interiorul ntreprinderilor.

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Pornindu-se de la realitatea c oamenii adaug valoare din valoarea pe care


o au, n cadrul procesului de implementare a sistemului de management al
siguranei sanitare a alimentului, au fost efectuate instruiri periodice ale
personalului, tocmai pentru a ridica nivelul de pregtire profesional. S-a dovedit
c aceast investiie uman este minim n termeni financiari dar foarte
important pentru ndeplinirea obiectivelor stabilite.
Implementarea conceptului de siguran sanitar a alimentelor. Punere
n practic a unui sistem de management care s garanteze sigurana produselor
alimentare ncepe ntotdeauna cu abordarea igienic a problemei (4). Patru puncte
sunt n mod particular importante: respectul legislaiei; luarea n considerare a
programelor prealabile (GMP); cunoaterea punctelor tari i punctelor slabe ale
ntreprinderilor n materie de inere sub control global a igienei; angajamentul i
motivarea ntregului personal. S-a dovedit c nu este oportun s se lanseze
demersul HACCP dac nu sunt ndeplinite toate prealabilele operaionale i de
infrastructur. Respectarea scrupuloas a bunelor practici de igien i de producie
(GMP) condiioneaz eficacitatea sistemului HACCP. Aceste bune practici de
includ ansamblul msurilor de igien definite pentru fiecare sector de producie.

CONCLUZII

Evoluia regulilor comerului i exigenele crescute ale consumatorilor au


fcut din sigurana alimentar o preocupare major a actorilor filierei alimentare,
inclusiv n ntreprinderi alimentare din Iasi. Sigurana alimentelor nu este
garantat dect atunci cnd responsabilitatea este partajat de toi membrii
ntreprinderii alimentare, de la managementul de vrf i operatori pn la
muncitori i chiar stagiari.
Pentru ca alimentul ce ajunge la masa consumatorului s fie propriu pentru
a fi mncat i c riscurile de contaminare s fie minimalizate, este absolut necesar
s fie implementat un sistem de proceduri i mecanisme de control. Deoarece nu
exist riscul zero, doar o bun implementare pe baza principiilor de baz a
respectrii igienei i bunelor practici de producie poate s garanteze alimente
care s nu afecteze sntatea celor care le consum.

BIBLIOGRAFIE

1. Autoritatea European de Siguran a Alimentelor: http://efsa.eu.int.


2. Autoritatea Naional Sanitar veterinar i pentru Sigurana Alimentelor - Legislaie
specific, http://www.ansv.ro/legislatie.php?cmd=index&tip=HOTA
3. Brdan Gh. - note de curs Sigurana alimentelor, USAMV Iasi (manuscris).
4. Codex Alimentarius CAC/RCP 1-1969, Rev. 4-2003. - Code d'usages international
recommande - principes gnraux d'hygine alimentaire,
http:// www.codexalimentarius.net
5. Comisia european - Sigurana alimentelor:
http://europa.eu.int/comm/food/index_fr.htm

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6. Livre blanc sur la securite alimentaire Commission des Communautes europeenes,


http://ec.europa.eu/dgs/h ealth_consumer/library/pub/pub06_fr.pdf.
7. Pachetul igien Regulamente ale Parlamentului european i ale Consiliului
(29.09.2004) http:// europa.eu.int/ eur-lex/ pri/ fr/ oj/ dat/ 2004/ l_226/
l_22620040625fr00030021.pdf
8. http://europa.eu.int/eur-lex/pri/fr/oj/dat/2004/l_226/l_22620040625fr00220082.pdf
9. http://eur-lex.europa.eu/LexUriServ/site/fr/oj/2004/l_226/l_22620040625fr00830127.pdf
10. http://eur-lex.europa.eu/LexUriServ/site/fr/oj/2004/l_165/l_16520040430fr00010141.pdf

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AGRICULTUR DURABIL I SECURITATE


ALIMENTAR N UNIUNEA EUROPEAN
PROGRAME DE ACIUNE ADOPTATE DE ROMNIA N
DOMENIUL AGRICULTURII DURABILE I A SIGURANEI
ALIMENTELOR
Teofilia BANU, O. LUCHIAN

n perioada actual, politica agricol a Uniunii Europene are ca obiective


prioritare asigurarea securitii alimentare i crearea condiiilor de punere n
practic a principiilor unei agriculturi durabile la nivel continental.
Reformele majore ntreprinse la nivel european n politica agricol, de
mediu i macroeconomic, a rilor dezvoltate sau a celor n curs de dezvoltare,
au creat condiii propice pentru promovarea unei agriculturi durabile i a
dezvoltrii rurale.
Scopul dezvoltrii agricole i rurale durabile este acela de a determina o
cretere susinut a produciei alimentare i de a asigura securitatea alimentar.
Crizele sanitare care s-au succedat n domeniul alimentar n ultimii ani au
diminuat ncrederea consumatorilor europeni n sigurana produselor alimentare.
Rspunsul autoritilor europene la aceast provocare s-a concretizat ntr-o
strategie global de asigurare a unui nivel ridicat de siguran alimentar. Aceasta
se bazeaz pe un control riguros al produselor alimentare i o supraveghere
permanent a drumului lor de la ferm pn pe masa consumatorilor.
Politica agricol la nivel european are n vedere gestionarea pe baze
tiinifice a riscurilor n domeniul alimentar, promovarea diversitii produselor
alimentare i a dreptului consumatorilor de a alege n cunotin de cauz.
n vederea integrrii n Uniunea European i a implementrii acquis ului
comunitar principalul instrument financiar de preaderare pentru agricultur i
dezvoltare rural de care dispune Romnia este Programul SAPARD.
Operaionalizarea acestuia n ara noastr se realizeaz prin Programul
Naional pentru Agricultur si Dezvoltare Rural, ce pune accent pe prioritile
identificate pentru perioada de pre-aderare, respectiv : mbuntirea structurilor
de prelucrare i marketing a produselor agricole i piscicole; controlul calitii
alimentelor; controlul veterinar i fito-sanitar; mbuntirea infrastructurii pentru
dezvoltare rural si agricultur; dezvoltarea economiei rurale si a resurselor
umane.
Programele elaborate de Autoritatea Naional Sanitar Veterinar i pentru
Sigurana Alimentelor au menirea s supravegheze implementarea practicilor
Uniunii Europene n domeniul siguranei alimentelor i s creeze condiiile pentru
adoptarea de reacii adecvate n situaii de criz pentru a asigura protejarea
intereselor i sntii cetenilor.

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CONSIDERAII ASUPRA CALITII IGIENICE A UNOR


PROBE DE LAPTE DESTINATE CONSUMULUI PUBLIC
CONSIDERATIONS REGARDING THE HYGIENIC QUALITY
OF SOME MILK SAMPLES FROM PUBLIC CONSUMPTION

Ioana DALEA, G. ZEGREAN


U.S.A.M.V. Cluj-Napoca

A verification of the hygienic quality of high temperature pasteurized


milk from sale was conducted, from A provider, in the way of which, the
following methods were applied: total mesophillic germ number
determination, impurities determination and somatic cell presence
determination. 20% of the 20 examined samples were situated outside the
maximum limit of consumption and sold like this in a market with a fridge
that didnt have any thermometers. These samples had a much higher total
germ number that the legal one (8.9 times higher). 10% of the samples had
impurities in the sediment and also somatic cells, and only 55% didnt
contain any mesophillic germs.

INTRODUCERE

Supravegherea continu i constant a calitii igienice a laptelui din


consum este deosebit de important pentru sntatea public. Un numr excesiv
de mare de germeni poate produce alterarea calitilor organoleptice ale laptelui
dar n acelai timp pericliteaz sntatea consumatorilor. Pe de alt parte, celulele
somatice pot adposti germeni patogeni, fiind ele nsele un indicator al eficienei
metodelor de igienizare a produsului. n laptele pasteurizat nalt teoretic, nu ar
trebui s mai existe celule somatice iar NTG m.a., conform normelor din ara
noastr, ar trebui s fie de maxim 300.000/ml. [1, 2, 3, 4]

MATERIAL I METOD

Au fost luate n lucru 20 de probe de lapte igienizat prin pasteurizare nalt,


procurate din comer, respectiv lapte de la furnizorul notat A, ambalat la cutie de
carton. S-a determinat numrul total de germeni mezofili aerobi (NTG m.a.), prin
obinerea de diluii succesive n ser fiziologic steril i nsmnare pe agar steril.
Citirea s-a fcut la 24 de ore, dup termostatare la 37C. De asemenea, laptele a
fost centrifugat la 3000 rpm timp de 10 minute, n vederea obinerii
lactosedimentului, din care s-au executat frotiuri colorate cu albastru de metilen i
Gram. Frotiuri s-au executat i din coloniile care au crescut n plcile
nsmnate, colorate prin metoda Gram.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

REZULTATE I DISCUII

Din cele 20 de probe luate n lucru, 5 se aflau n afara limitei maxime de


consum i comercializate astfel (20% dintre probe), din care 3 probe cu dou zile,
iar restul, cu trei zile. Din punct de vedere organoleptic, doar probele care
depiser limita maxim de consum cu 3 zile, au prezentat miros i gust acid.
n cazul celor 15 probe care se situau n perioada de consum, NTG m.a. s-a
situat n limitele admise n ara noastr, respectiv sub 300.000 germeni/ml lapte.
55% dintre probe nu prezentau germeni mezofili aerobi.
n cazul probelor cu termenul limit de consum depit, valorile NTG m.a.
se situau cu mult peste limita admis, valorile fiind prezentate n tabelul 1. Flora
microbian prezent n toate probele a constat stafilococi i streptococi Gram
pozitivi, cocobacili Gram negativi i bacili Gram pozitivi i negativi. Impuriti i
celule somatice au fost prezente doar n dou dintre probe (10%), acestea din
urm fiind reprezentate de neutrofile, limfocite i macrofage activate (Fig 1, 2), n
numr mic, n medie 1020 celule pe frotiu. Sedimentul acestor probe a avut o
culoare glbuie, cu impuriti la examinarea cu obiectivul x40 i era cantitativ
crescut fa de al celorlalte probe de lapte.
Tabelul 1
Parametrii calitii igienice a probelor de lapte luate n lucru destinate consumului public

Proba NTG m.a./ml Celule somatice Impuriti


1 - - -
2 23.000 Prezente Prezente
3 - - -
4 4.000 Prezente Prezente
5 1.000 - -
6 - - -
7 - - -
8 - - -
9 - - -
10 - - -
11 82.000 - -
12 - - -
13 - - -
14 - - -
15 - - -
16* 1.230.000 - -
17* 480.000 - -
18** 2.140.000 - -
19* 749.000 - -
20** 2.680.000 - -
* n afara limitei de consum (2 zile)
** n afara limitei de consum (3 zile)

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Este de menionat faptul c vitrina frigorific a unitii de comercializare a


produselor lactate nu era prevzut cu un sistem de monitorizare electronic
vizibil a temperaturii.

Fig. 1. Neutrofile i macrofage n laptele Fig. 2. Neutrofile n laptele de la furnizorul


de la furnizorul A, coloraie cu albastru A, coloraie cu albastru de metilen,
de metilen, x1000 x1000

CONCLUZII

Deoarece s-a constatat prezena celulelor somatice i mai ales a


impuritilor n laptele pasteurizat nalt (10% din probe), este evident c metodele
de igienizare, respectiv filtrarea i pasteurizarea nu sunt supravegheate constant i
corespunztor n cazul furnizorului A.
Pstrarea necorespunztoare i peste limita menionat pe ambalaj, duce la
multiplicarea excesiv a germenilor mezofili aerobi cu mult peste limitele
admise (chiar de aproape 10 ori n cazul probei 20), care produc alterri de gust i
miros ale laptelui.
Exist uniti care comercializeaz lapte i produse lactate pstrate n
condiii precare de temperatur i care nu in cont de recomandrile
productorilor, nscrise pe ambalaj.
Concluzia clar care se desprinde din datele obinute este aceea c
supravegherea calitii igienice a laptelui destinat consumului public necesit
mbuntiri, ncepnd de la procesarea primar a laptelui pn la desfacere.

BIBLIOGRAFIE

1. http://www.promediapublishing.it/CMS/PDF/articoli/Art_pdf_164_DossierLatte
2. Rotaru O., Gu Camelia, M. Mihaiu, 1999 - Controlul sntii produselor de origine
animal, Editura Hipparion, Cluj-Napoca
3. Rotaru O., M. Mihaiu, 2004 - Igiena veterinar a produselor alimentare, Vol 2,
Editura Risoprint, Cluj-Napoca
4. The Raw Milk: International Journal of Food Sciences and Nutrition, Volume 52, nov
2001

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INFLUENA TEMPERATURII ASUPRA CRETERII


TULPINILOR DE BACILLUS CEREUS IZOLATE
DIN ALIMENTE
TEMPERATURE INFLUENCE ON GROWTH CHARACTERISTICS
OF FOOD ISOLATES OF BACILLUS CEREUS

V. FLORITEAN1, Elena IAN2,


C. SRBU2, M. CARP CRARE1
1
U.S.A.M.V. Iai
2
L.S.V. Iai

Refrigerated foods have a relevant potential in the food market,


although the potential risk posed by sporulated emerging psycrotrophs
pathogens has to be evaluated. Bacillus cereus is one of these pathogenic
microorganisms. In this paper, the ability to grow of several food isolates of
Bacillus cereus was evaluated.
Twenty strains of Bacillus cereus isolated from milk and meat
products have been used to study growth characteristics on solid and liquid
media in detail. The analysis revealed a large biodiversity within the
Bacillus cereus food strains. Fourteen strains were able to grow at 7C and
could be considered as psychrotolerant. All the milk and dairy isolates
could grow at 7C. However, none of these strains could grow at 48C.
Only five of the strains (20%) have grown at 48C. All strains grew well at
20C and 42C.

Key words: Bacillus cereus, Psychrotolerant, Growth

Bacillus cereus este un microorganism sporulat, Gram pozitiv aerob, larg


rspndit n mediu cunoscut att pentru aciunea s-a spoliatoare ct mai ales
pentru capacitatea acestuia de a produce, n anumite condiii toxiinfecii
alimentare.
Bacillus cereus produce dou tipuri de toxiinfecii alimentare sindromul
diareagen i cel vomitiv denumite astfel dup manifestrile clinice
predominante ale celor dou entiti. (7, 8)
Abilitatea de a sporula face ca bacteria s reziste tratamentelor termice
uzuale aplicate alimentelor n procesul tehnologic de obinere iar prezena n
cadrul speciei a tulpinilor psichrotolerante permite acesteia s se dezvolte la
temperatura de refrigerare. Aceste caracteristici fiziologice permit supravieuirea
i chiar dezvoltarea bacteriei n alimente mai ales n cazul alimentelor
semipreparate ceea ce constituie un risc permanent pentru consumatori i o
preocupare permanent a specialitilor din industria alimentar pentru a se reduce
pierderile. (1, 5, 6, 9)

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Lucrarea de fa investigheaz abilitatea de a se dezvolta a unor tulpini de


Bacillus cereus izolate din alimente de origine animal pe medii solide i lichide
la diferite temperaturi. Cunoaterea comportamentului tulpinilor de Bacillus
cereus cu origine alimentar sub aciunea unor factori de mediu este important
fiind util att n evaluarea riscului prezenei bacteriei n alimente ct i n
stabilirea msurilor adecvate pentru reducerea incidenei acestuia n alimente.

MATERIAL I METOD

Influena temperaturii asupra creterii tulpinilor din specia Bacillus cereus a


fost testat pe un numr de 20 tulpini de Bacillus cereus izolate din produse
alimentare de origine animal. Testarea s-a efectuat att pe mediu solid ct i n
mediu lichid.
Pe mediul solid. Pentru a determina capacitatea de dezvoltare la diferite
temperaturi tulpinile izolate au fost cultivate iniial pe bulion tripton-soia (TSB),
incubate la 30C pentru 24 ore. Din culturile tinere un inocul de 0,1 ml s-a
nsmnat, prin striere, pe agar tripton-soia (TSA).
Plcile nsmnate au fost incubate la temperaturi diferite (7, 20, 42 i
48C) timp de 6 zile.
Plcile au fost examinate zilnic pentru a se depista momentul apariiei
coloniilor vizibile pe mediu. Coloniile dezvoltate au fost confirmate conform
protocolului impus prin SR ISO 7932/1997.
n mediul lichid. Pentru o mai bun caracterizare a creterii tulpinilor de
Bacillus cereus sub influena temperaturii s-au efectuat i teste pe medii lichide.
Tulpinile testate au fost cultivate iniial pe bulion creier cord (BHI) i
incubate timp de 24 ore la 30C. Pentru fiecare tulpin testat s-au pregtit cte
patru baloane Erlemeyer cu cte 30 ml bulion creier-cord i cte patru eprubete
cu cte 10 ml bulion creier-cord, toate fiind nsmnate cu cultur tnr de
Bacillus cereus pn la aprox. 103 104 UFC/ml.
Pentru testarea creterii la temperatur sczut, baloanele au fost
incubate, cte dou, la temperatura de 7C timp de 14 zile i la 20C timp de 5
zile. Monitorizarea creterii s-a efectuat prin metoda diluiilor seriate la intervale
de timp variabile (n funcie de temperatura de incubare). Diluiile seriate s-au
preparat pe ap peptonat salin, din fiecare diluie fiind prelevat cte 0,1 ml care
cu care s-a nsmnat prin striere plci Petri cu agar creier-cord care au fost
incubate la 30C timp de 24 ore.
Pentru testarea creterii la temperatur ridicat eprubetele inoculate cu
cultur tnr i incubate la 42C timp de 48 ore i la 48C timp de 10 zile.
Monitorizarea creterii s-a fcut prin metoda diluiilor seriate la intervale de timp
variabile.
Curbele de cretere au fost generate pe baza rezultatelor obinute cu
ajutorul unui program statistic folosindu-se o funcie Gompertz modificat
recomandat i/sau utilizat i de ctre ali autori (3, 10, 11, 12, 13).
Pentru caracterizarea creterii tulpinilor de Bacillus cereus s-a estimat
timpul de generaie, rata de cretere i durata fazei de lag folosindu-se parametrii
matematici (A,B,C,M) calculai n prima etap.

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Y= A + C*EXP(-EXP(-B*(X-M)))
n care:
Y - populaia bacterian la momentul X (log UFC/ml)
X - timpul (ore)
A - inoculul iniial (log UFC/ml)
B - rata de cretere relativ la momentul M (generaii/or)
C - maximul statistic atins de populaia bacterian (log UFC/ml)
M - momentul la care se atinge creterea maxim (ore)
RCE (Rata de cretere exponenial) (log UFC/ml/or) = B*C/e
DFL (Durata fazei de lag) (ore) = M-(1/B)
TG (Timp de generaie) (ore) = (log102)e/B*C

REZULTATE I DISCUII

Rezultatele obinute sunt prezentate n tabelele 1-3 i fig. 1.


Pe mediu solid la 70C tulpinile de Bacillus cereus s-au dezvoltat lent,
coloniile fiind vizibile ncepnd cu ziua a treia de incubaie. Din cele 20 tulpini
testate doar 14 au fost capabile s se dezvolte, n condiii care s conduc la
colonii vizibile n cele 6 zile ct a durat experimentul. La temperatura de 20C
toate tulpinile testate s-au dezvoltat dnd colonii vizibile nc din primele 24 ore
de incubaie. La 42C creterea s-a produs n bune condiii, toate tulpinile au dat
colonii vizibile dup dou zile de incubaie. Temperatura ridicat s-a dovedit a fi
un factor care a inhibat dezvoltarea unui numr mare din tulpinile testate (15), iar
la tulpinile la care s-au evideniat colonii, acestea au aprut dup 48 de ore de
incubaie (2 tulpini) i dup 72 de ore (3 tulpini). (Tabelul 1)
Tabelul 1
Dezvoltarea tulpinilor de Bacillus cereus pe medii solide la diferite temperaturi

Nr. de tulpini care au dat colonii


Tulpini testate 20 din care:
Temperatura vizibile/timp (ore)
C nu au
24 48 72 96 120 144 au crescut
crescut
7 - - 2 9 3 - 14 6
20 20 - - - - - 20 -
42 16 6 - - - - 20 -
48 - 2 3 - - - 5 15

n ceea ce privete comportamentul tulpinilor de Bacillus cereus n funcie


de originea acestora se poate observa o predominan a tulpinilor care au
abilitatea de a se dezvolta la temperaturi sczute (psichrotolerante) n lapte i
produse lactate n vreme ce tulpinile izolate din carne i produse din carne s-au
dezvoltat mai bine la temperaturi ridicate, fiind singurele tulpini care s-au
dezvoltat la 48C. (Tabelul 2)
Pe medii lichide au fost testate doar tulpinile la care s-a evideniat cretere
pe mediul solid. Pe mediul lichid creterea a fost mai rapid, ncepnd dup o

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perioad de lag medie cuprins ntre 1,037 ore i 74,53 ore. Cea mai lung
perioad medie de lag a fost constatat la temperatura de 7C iar cea mai scurt la
temperatura de 42C. Comparativ cu tulpinile izolate din carne, cele din lapte s-
au dezvoltat bine la temperatura de 7C dar mult mai lent la temperatura de 42C.
Tabelul 2
Dezvoltarea tulpinilor de Bacillus cereus pe medii solide la diferite temperaturi n
funcie de originea lor

Tulpini dezvoltate/origine
Temperatura Carne i produse din
Lapte i produse lactate
C carne
Total (n =10)
(n = 10)
Nr. % Nr. %
7 14 10 100 4 40
20 20 10 100 10 100
42 20 10 100 10 100
48 5 - - 5 50

La temperatura de 20C toate tulpinile, indiferent de origine s-au dezvoltat


foarte bine ceea ce nseamn c meninerea la temperatura camerei a alimentelor
contaminate cu Bacillus cereus constituie un risc, alimentele fiind expuse
degradrii rapide iar n cazul tulpinilor toxigene apare riscul apariiei
toxiinfeciilor alimentare la consumatori.
Cea mai rapid cretere s-a estimat la tulpinile din carne la 42C urmate de
tulpinile din lapte la 20C i respectiv 42C, tulpinile din carne la 20C.
La 48C pe mediul solid tulpinile au nceput creterea dup o faz de lag
relativ scurt (aprox. 22 ore) i au ajuns la faza staionar n timp scurt, rata de
cretere exponenial fiind destul de ridicat.

Fig. 1 Comportamentul tulpinilor de Bacillus cereus la diferite temperaturi n funcie de


originea lor

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Nivelul maxim atins de tulpinile de Bacillus cereus n condiiile studiului


au fost cuprinse ntre 107 i 109 fiind dependent, n principal, de temperatura de
incubaie, de tulpina bacterian i mai puin cu dimensiunea inocului iniial.
(Tabelul 3 i Fig. 1)
Rezultatele obinute sunt comparabile cu cele publicate de ali autori dar cu
unele diferene generate de condiiile n care s-au desfurat experimentele i de
modelul matematic utilizat pentru prelucrarea datelor. De asemenea, n
interpretarea rezultatelor obinute pe medii lichide trebuie s inem cont de faptul
c metoda folosit (funcia Gompertz modificat), dei elocvent n caracterizarea
creterii bacteriene, are tendina de a supraestima rata de cretere exponenial (cu
pn la 15% dup unii autori). (2, 4, 11, 12)

CONCLUZII

Rezultatele obinute reflect marea biodiversitate care se manifest n


cadrul speciei Bacillus cereus, tulpinile cu origine alimentar caracterizate prin
acest studiu fiind capabile s se dezvolte ntr-un interval larg de temperatur;
Tulpinile care au abilitatea s se dezvolte la temperaturi sczute (7C) au
fost evideniate cu o frecven mai mare n lapte i produse din lapte, ceea ce
denot posibilitatea unei selecii a acestor tulpini pe parcursul procesului
tehnologic de prelucrare i depozitare a laptelui i a produselor lactate;
Tulpinile izolate din carne s-au dezvoltat mai bine la temperaturi medii i
ridicate n comparaie cu tulpinile din lapte. Prezena bacteriei n carne i produse
din carne este rezultatul contaminrii cu tulpini din mediu iar n cazul produselor
din carne i cu tulpini de origine vegetal cunoscute pentru abilitatea lor de a
crete la temperaturi ridicate;
Dei au origine comun, n sol, tulpinile de Bacillus cereus din alimente
manifest diferene apreciabile n modul de reacie la aciunea factorilor de mediu
i aceasta datorit faptului c aceste tulpini se adapteaz, n funcie de specificul
fiecrui aliment, la anumite condiii supravieuind n/pe instalaiile din industria
alimentar i constituie, astfel, surse perpetue de contaminare a alimentelor.
Abilitatea bacteriilor din specia Bacillus cereus de a sporula, combinat cu
adaptabilitatea mare a formelor vegetative la cele mai variate nie ecologice fac
din aceast specie o problem pentru industria alimentar i pentru sigurana
alimentar n special.
Chiar dac datele din prezenta lucrare au la baz doar unul din multitudinea
de factori care influeneaz creterea bacteriilor din specia Bacillus cereus n
alimente, astfel de modele sunt utile pentru evaluarea riscului reprezentat de
prezena acestei bacterii n alimente i n stabilirea unor msuri eficiente prin care
s se previn atingerea unui numr de bacterii capabil s produc tulburri la
consumatori.

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BIBLIOGRAFIE

1. Andersson, A., Rnner, U., Granum, P.E., 1995 - What problems does the food
industry have with the spore-forming pathogens Bacillus cereus and Clostridium
perfringens? International Journal of Food Microbiology 28: 145-155
2. Benedict, R.C., Partridge, T., Wells, D., Buchanan, R.L., 1993 - Bacillus cereus:
aerobic growth kinetics. Journal of Food Protection 56: 211214.
3. Chorin, E., Thuault, D., Cleret, J.J., Bourgeois, C.M. (1997) Modelling Bacillus
cereus growth. International Journal of food Microbiology 38: 229234.
4. Delignette-Muller M.L., Rosso L., 2000 - Biological variability and exposure
assessment. International Journal of Food Microbiology 58: 203212
5. Dufrenne, J., Soentoro, P., Tatini, S., Day, T., Notermans, S., 1994 -
Characteristics of Bacillus cereus related to safe food production. International
Journal of Food Microbiology 23: 99-109.
6. Korn, R., 1981 - Incidena speciei Bacillus cereus n unele alimente de origine
animal i proprietile tulpinilor izolate. Tez de doctorat, I.A.N.B., Bucureti
7. Kramer, J.M., Gilbert, R.J., 1989 - Bacillus cereus and other Bacillus species.
Foodborne Bacterial Pathogens, Ed. by M. P. Doyle, Marcel Dekker,INC. New
York and Basel 21-70
8. McKillip, J.L., 2000 - Prevalence and expression of enterotoxins in Bacillus cereus
and other Bacillus spp., a literature review. Antonie van Leeuwenhoek 77, 393-
399
9. Mureanu, I., 2000 - Cercetri privind incidena n unele alimente de origine animal
i particularitile morfologice ale speciei Bacillus cereus, potenial patogen
pentru om, n zona central a Transilvaniei. Tez de doctorat U..A.M.V.
Bucureti
10. Pinon, A., Zwietering, M., Perrier, L., Membr, J.M., Leporq, B., Mettler, E.,
Thuault, D., Coroller, L., Stahl, V., Vialette, M., 2004 - Development and
Validation of Experimental Protocols for Use of Cardinal Models for Prediction of
Microorganism Growth in Food Product. Applied and Environmental
Microbiology, 70:1081-1087
11. Rajkowski, K.T., Mikolajcik, E.M., 1987 - Characteristics of selected strains of
Bacillus cereus. Journal of Food Protection 50: 199-205.
12. Zaika, L.L. Scullen, O.J., 1996 - Growth of Shigella flexneri in foods: comparison of
observed and predicted growth kinetics parameters, International Journal of
Food Microbiology, 32: 91-102
13. Zwietering, M.H., De Koos, J.T., Hasenack, B.E., De Witt, J.C., Van'T Riet, K.,
1991 - Modeling of bacterial growth as a function of temperature. Applied
Environmental Microbiology, 57: 1094- 1101

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IDENTIFICAREA PRINCIPALELOR SURSE


DE CONTAMINARE CU MICROORGANISME
A CARCASELOR DE PASRE N ABATOR
IDENTIFICATION OF THE MAJOR SOURCES OF MICROBIAL CONTAMINATION
OF CHIKEN CARCASSES IN ABATTOIRS

Carmen CREU, V. FLORITEAN,


M. CARP-CRARE
U.S.A.M.V. Iai

The study was carried out in a poultry slaughterhouse from Iai


county. A total number of 270 swab samples were taken from different sites
of 54 poultry carcasses in order to evaluate the microbiological status and
to identify the most common sources of carcass contamination during
various unit operations. The total viable counts and two important indicator
microorganisms (coliforms and Escherichia coli) were monitored.
According to the region of carcasses the total viable counts varied between:
6.2-6.6 (log cfu/cm2) after bleeding, 3.8-4.5 after scalding, defeathering
and washing, 5.3-6.1 after evisceration, 4.35.1 after washing and 4.14.8
after chilling. The indicator microorganisms (coliforms and E. coli)
prevalence was quite high. Legs showed the maximum level of
contamination and high prevalence of indicator microorganisms being one
of the most critical carcass region. Proper monitoring of each step in
poultry slaughterhouse processing it is important in order to establish
control measures in meat quality assurance process.

Key words: poultry, poultry microbiology, coliforms, E. coli, poultry quality

Consumul de carne de pasre, datorit valorii nutritive i a digestibilitii


ridicate, este ntr-o continu cretere. La ora actual exist o preocupare continu
pentru a se asigura calitatea crnii de pasre, prin controlul strii de sntate a
psrilor destinate sacrificrii i prin mbuntirea metodelor de control a strii
de igien n abatoarele de psri (implementarea HACCP). Carnea de pasre
sufer pe parcursul procesrii manipulri multiple, contaminarea fiind posibil pe
tot fluxul tehnologic. Sursele microorganismelor contaminante sunt diverse i
numeroase, cea mai important fiind psrile nsi, care pot purta n special n
tractul digestiv o microflor bogat i diversificat (1, 2, 3, 8, 9, 11).
Prezena pe carcasele de pasre, n numr mare, a unor microorganisme,
indic lipsa de igien pe parcursul procesrii n abator sau o serie de deficiene
legate de transportul i/sau depozitarea crnii (4, 5 , 6, 7 ,11).
Prezentul studiu i propune s evalueze, prin intermediul unor
microorganisme indicator al strii de igien, nivelul contaminrii cu
microorganisme n diferite stadii ale prelucrrii psrilor n abator i s identifice,

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n acest fel, etapele unde calitatea microbiologic a crnii de pasre poate fi


ameliorat prin stabilirea prin msuri corective adecvate.

MATERIAL SI METODA

Au fost examinate un numr de 270 de probe. Probele au fost recoltate de


pe fluxul tehnologic de la un abator din judeul Iai dup cum urmeaz: dup
sngerare, dup oprire, deplumare i splare, dup eviscerare, dup splare i
dup rcire. Probele s-au recoltat cu ajutorul unor tampoane de vat sterile
umezite cu ap peptonat steril, prin tergerea repetat (timp de 15-30 sec.) a
unei suprafee de aprox. 100 cm2 din diferite zone ale carcasei (picioare, piept i
aripi). Tampoanele de vat au fost introduse n 10 ml ser fiziologic peptonat 1%,
n tuburi cu capac sterile. Tuburile au fost transportate la laborator n condiii
corespunztoare.
Tuburile au fost agitate 30 sec. cu ajutorul unui Vortex dup care din serul
fiziologic peptonat s-au preparat diluii seriate pn la 10-6.
Numrul total de germeni aerobi mezofili a fost determinat pentru diluiile
10-4 i 10 -5 pe agar nutritiv prin nglobarea a 0,1 ml inocul. Pentru fiecare diluie s-
au efectuat cte dou plci. Plcile nsmnate au fost incubate la 37C timp de
24 - 48 ore.
Pentru determinarea microorganismelor indicator s-au folosit medii
selective specifice. Pentru determinarea numrului probabil de bacterii coliforme
s-au efectuat diluii zecimale, din fiecare diluie introducndu-se cte 1 ml n cte
3 eprubete cu mediu BBLV (bulion lactozat cu sruri biliare i verde briliant) i tub
Durham. S-a incubat 24 48 ore la 37 C. Interpretarea s-a fcut dup producia
de gaz i calcularea mediei de 3 cifre (funcie de nota primit de fiecare din cele 3
eprubete cu mediul BBLV), media obinut fiind interpretat cu ajutorul tabelului
Mac Grady.
n paralel s-a utilizat i metoda de determinare a numrului de bacterii
coliforme prin numrarea coloniilor (ISO 4832). Din diluiile zecimale se
repartizeaz cu pipete sterile cte 1 ml n cte 2 plci Petri. n fiecare plac se
toarn mediu VRBL (geloz cu lactoz sruri biliare cristal violet i indicator) topit
i rcit la 45 C. Plcile pregtite se incubeaz la 35 C timp de 24 ore. Dup
incubare se numr coloniile rou violacee i se calculeaz media cu formula: N
= c / (n1 + 0,1 n2) d [n care c reprezint suma coloniilor numrate; n1 este
numrul de cutii de la prima diluie reinute pentru numrare; n2 este numrul de
cutii de la a doua diluie reinute pentru numrare; d este rata diluiei care
corespunde primei diluii folosite]
Pentru identificarea bacteriei Escherichia coli STAS ISO 4832, pentru
determinarea numrului probabil de E. coli SR ISO 7251 Conform acestui
standard se nsmneaz serii de cte trei eprubete, coninnd mediu de
mbogire selectiv i tuburi Durham, cu proba de analizat i cu diluiile decimale
ale acesteia. Acest mediu este reprezentat de bulion cu tripton i lauril sulfat.
Eprubetele astfel pregtite se incubeaz la 35C sau 37C, pentru 24 h - 48 h.
Dup acest interval, din fiecare eprubet n care s-a observat degajare de
gaz, se nsmneaz cel de-al doilea mediu de mbogire selectiv, respectiv

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bulionul E.C. Eprubetele se incubeaz la 45C pentru 24 h - 48 h. n continuare,


din fiecare cultur care a prezentat degajare de gaz se nsmneaz cte o
eprubet cu ap triptonat care se incubeaz la 45C, pentru 48 h.
Dup incubarea eprubetelor cu ap triptonat, n fiecare se adaug 0,5 ml
reactiv Kovacs pentru identificarea prezenei indolului. Se consider reacie
pozitiv dac la suprafaa culturii apare un inel rou. Calculul numrului cel mai
probabil de E. coli se realizeaz, n funcie de numrul eprubetelor care conin
cultur considerat pozitiv, folosind tabelul Mac Grady.

REZULTATE I DISCUII

Prezena microorganismelor pe i n carne poate fi n anumite condiii


periculoas pentru consumator. Evaluarea microflorei crnii, calitativ i cantitativ,
precum i identificarea etapelor fluxului tehnologic care prin specificul acestora
pot contribui la modificarea acesteia este important n stabilirea msurilor
preventive i corective prin intermediul crora evoluia microflorei poate fi
controlat.
Tabelul 1
2
Valorile NTG/cm pe diferite regiuni ale carcaselor de pasre pe parcursul fluxului
tehnologic de prelucrare n abator

2
NTG (log UFC/cm eroarea standard)
Etapa Regiunea Nr. probe
tehnologic carcasei examinate Valoare Valoare
Media
minim maxim
Aripi 54 5,2 6,8 6,2 0,19
Sngerare Picioare 54 5,5 6,5 6,3 0,13
Piept 54 5,8 6,8 6,6 0,23
Aripi 54 3,4 5,1 3,8 0,12
Oprire -
Picioare 54 3,8 5,6 4,5 0,21
deplumare
Piept 54 3,6 5,8 4,2 0,14
Aripi 54 4,5 6,3 5,3 0.20
Eviscerare Picioare 54 5,6 7,1 6,1 0,17
Piept 54 5,3 6,8 5,5 0,26
Aripi 54 3,1 5,1 4,3 0,22
Splare Picioare 54 4,2 6,3 5,1 0,20
Piept 54 4,1 5,3 4,8 0,04
Aripi 54 3,1 4,5 4,1 0,19
Rcire Picioare 54 3,8 5,2 4,8 0,32
Piept 54 3,7 4,8 4,5 0,27

Numrul total de germeni (NTG), determinat pe parcursul fluxului


tehnologic al prelucrrii psrilor n abator, a furnizat valori cuprinse ntre 3,1
0,1 log ufc/cm2 i 7,1 0,02 log ufc/g. Valorile cele mai mari au fost depistate
dup efectuarea sngerrii i a eviscerrii, la polul opus, valorile cele mai mici
fiind obinute dup oprire i n cursul etapelor finale ale prelucrrii, dup splare

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i respectiv dup rcirea carcaselor. Diferene au fost constatate i n cazul celor


trei regiuni examinate, astfel, NTG a avut valorile cele mai mari n mod constant
pe suprafaa picioarelor i n anumite situaii pe suprafaa pieptului (Tabelul 1).
Valorile obinute sunt asemntoare cu cele publicate, n literatura de
specialitate, de ali autori (5, 10, 11).
Valorile mari ale NTG dup sngerare sunt consecina unei ncrcturii
mari de microorganisme cu care ajung psrile n abatoare iar dup eviscerare
valorile mari ale NTG se explic prin specificul acestei operaiuni cnd carcasa
poate veni n contact cu masa gastro-intestinal i chiar cu coninutul acesteia,
situaie care este mai frecvent atunci cnd operaiunea se efectueaz defectuos.
Coliformii i E. coli, doi parametri microbiologici utilizai n industria
alimentar pentru aprecierea strii de igien a produselor alimentare i a spaiilor
tehnologice, au fost identificai la 24,8% respectiv 18,57% din carcasele
examinate.
Valorile obinute pentru coliformi au fost cuprinse ntre 2,04 i 3,98 log
cfu/cm2, distribuia acestora fiind strns corelat cu cea evideniat n cazul
valorilor NTG. n ceea ce privete regiunile corporale examinate, coliformii au
fost prezeni n mod constant pe suprafaa picioarelor, tot aici nregistrndu-se i
valorile cele mai mari (Tabel 2).
Tabelul 2
2
Valorile indicatorului coliformi/cm pe diferite regiuni ale carcaselor de pasre
pe parcursul fluxului tehnologic de prelucrare n abator

Probe
Etapa Regiunea Nr. Probe pozitive
negative Media*
tehnologic carcasei probe
Nr. % Nr. %
Aripi 54 33 61,1 21 38,8 3,69 0,12
Sngerare Picioare 54 36 66,6 18 33,3 2,91 0,16
Piept 54 21 38,8 33 61,1 2,85 0,24
Aripi 54 - - 54 100 -
Oprire -
Picioare 54 48 88,8 6 11,2 2,42 0,44
deplumare
Piept 54 45 83,3 9 16,7 2,21 0,26
Aripi 54 27 50,0 27 50 3,3 0,23
Eviscerare Picioare 54 36 66,6 18 33,3 3,9 1,04
Piept 54 36 66,6 18 33,3 3,2 0,52
Aripi 54 - - 54 100 -
Splare Picioare 54 18 33,3 36 66,6 2,11 0,32
Piept 54 9 16,7 45 83,3 1,86 0,46
Aripi 54 - - 54 100 -
Rcire Picioare 54 6 11,1 48 88,8 1,75 0,19
Piept 54 - - 54 100 -
* log UFC/cm2 eroarea standard

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Escherichia coli, specie ncadrat n categoria coliformilor, care cuprinde i


tulpini cu potenial patogen, a fost identificat mai frecvent pe suprafaa
picioarelor i a aripilor imediat dup eviscerare. Valorile obinute s-au ncadrat
ntre 2,03 i 3,1 log ufc/cm2 fiind n marea majoritate a cazurilor cu valorile
coliformilor (Tabelul 3).
Tabelul 3
Valorile indicatorului E. coli/cm2 pe diferite regiuni ale carcaselor de pasre pe
parcursul fluxului tehnologic de prelucrare n abator

Probe
Etapa Regiunea Nr. Probe pozitive
negative Media*
tehnologic carcasei probe
Nr. % Nr. %
Aripi 54 9 16,7 45 83,3 1,23 0,18
Sngerare Picioare 54 9 16,7 45 83,3 1,51 0,12
Piept 54 9 16,7 45 83,3 1,31 0,42
Aripi 54 - - 54 100 -
Oprire -
Picioare 54 6 11,1 48 88,8 1,12 1,41
deplumare
Piept 54 - - 54 100 -
Aripi 54 18 33,3 36 66,6 1,40 0,22
Eviscerare Picioare 54 36 66,6 18 33,3 1,82 0,63
Piept 54 6 11,1 48 88,8 1,33 0,09
Aripi 54 - - 54 100 -
Splare Picioare 54 9 16,7 45 83,3 1,10 0,10
Piept 54 6 11,1 48 88,8 1,02 0,09
Aripi 54 - - 54 100 -
Rcire Picioare 54 3 5,5 51 94,5 1,12 0,02
Piept 54 - - 18 100 -
2
* log UFC/cm eroarea standard

Rezultatele obinute n cazul celor doi indicatori - coliformi i E. coli -


relev un procent uor mai ridicat de probe pozitive dect cele publicate de ali
autori. Acest fapt se explic prin condiiile de prelucrare i folosirea unor utilaje
mai puin performante n abatorizarea psrilor.

CONCLUZII

Studiul a relevat c din punct de vedere microbiologic etapele care


contribuie cel mai mult la contaminarea carcaselor de pasre n abatorul sunt
sngerarea i eviscerarea, etape care necesit o atenie deosebit pentru a reduce
gradul de contaminare a carcaselor n abatoare.
Dintre regiunile corporale analizate, picioarele reprezint regiunea cu cele
mai mari deficiene din punct de vedere microbiologic, furniznd cel mai des
rezultate pozitive la parametrii examinai.

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Rezultatele obinute nu au permis stabilirea unei corelaii pozitive constante


ntre NTG i cei doi indicatori ai strii de igien (coliformi i E. coli).
Stabilirea unor msuri, pornind de la instruirea permanent a personalului
din abatoare privind punctele critice, aplicarea unor tehnologii performante i
ntreinerea n stare bun de igien i de funcionare a instalaiilor i a spaiilor
sunt absolut necesare pentru reducerea contaminrii carcasei i mbuntirea
calitii microbiologice a crnii de pasre.

BIBLIOGRAFIE

1. Anand, S.K., Mahapatra, C.M., Pandey, N.K., Verma, S.S., 1989 - Microbial changes
on chicken carcasses during processing. India Journal of Poultry Sciences 24:
203-209.
2. Brzoi, D., Apostu, S., 2002 - Microbiologia produselor alimentare Ed. RISOPRINT,
Cluj-Napoca, 124-133.
3. Berrang, M.E., Ladley, S.R., Buhr, R.J., 2001 - Presence and level of
Campylobacter, coliforms, E. coli and total aerobic bacteria from broiler parts
with and without skin. Journal of Food Protection 64: 184-188.
4. Carp-Crare M., indilar E., Guguianu Eleonora, 1991 - Unele aspecte privind
portajul cu Yersinia enterocolitica i Campylobacter jejuni la gin. Lucr.
tiinifice U..A.M.V. Iai, 33-34: 156-159.
5. Kotula, K.L., Pandya, Y., 1995 - Bacterial contamination of broiler chickens before
scalding. Journal of Food Protection 58: 1326-1329.
6. indilar E., Grecianu Al., Verde Elena, Carp-Crare M., 1988 - Izolarea i
identificarea microflorei psichrotrofe de pe carcasele de pasre conservate prin
frig. Lucr. tiinifice U..A.M.V.Iai, 31: 81-83.
7. indilar E., Grecianu Al., Carp-Crare M., Negrea A., Bondoc I., 1994 - Unele
aspecte privind reducerea gradului de poluare microbian a crnii de pasre,
Lucr. tiinifice, U..A.M.V.Iai, 37: 101-106.
8. indilar E., Bondoc I., Moroan .Z., indilar E.V., 1997 - Unele aspecte privind
sursele de poluare microbian a carcaselor de pui broiler I, Lucr. tiinifice
U..A.M.V.Iai, 40: 80-93.
9. indilar E., Bondoc I., indilar E.V., 1997 - Unele aspecte privind sursele de poluare
microbian a carcaselor de pui broiler II, Lucr. tiinifice U..A.M.V.Iai, 40, 94-
99.
10. Vayda, V.M., Paturkar, A.M., Waskar, V.S., Zende, R.J., Rawool, D.B., 2005 -
Detection of indicator microorganisms on poultry carcass sites in an organized
slaughterhouse. Journal of Muscle Foods 16: 289-297.
11. Yashoda, K.P., Sachindra, N.M., Sakhare, P.Z., Rao, D.N., 2001 - Microbiological
quality of broiler chicken carcass processed hygienically in a small scale poultry
processing unit. Journal of Food Quality 24: 249-259.

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APLICAREA SISTEMULUI HACCP NTR-UN ABATOR DE


PORCI DIN REGIUNEA DE NORD-EST A ROMANIEI
IMPLEMENTATION OF THE HACCP SYSTEM
IN THE PORK SLAUGHTERING HOUSE IN NORD-EAST REGION OF ROMANIA

Gh. BRDAN, B. PSRIN


U.S.A.M.V. Iasi

From November 2005 to July 2006, the hazard analysis, critical


control points (HACCP) system was implemented to the benefit of the swine
abattoir in Nord-Est region of Romania. The technological process was
initially analyzed and the samples for bacteria indicators were realized.
Microbiological analyses revealed a poor sanitary quality of samples from
all the production lines tested. Implementation of HACCP method resulted
in reduction of microbiological contamination, but this decrease was not
significant compared with other steps of process. This method, which in
effect seeks to plan out unsafe practices, differs from traditional "produce
and test" quality assurance methods which are less successful and
inappropriate for highly perishable foods.

Key worlds: HACCP, pig, slaughtering house, management system.

INTRODUCERE

Politica siguranei alimentelor din cadrul Uniunii Europene ia n


considerare ntregul lan al alimentelor/furajelor destinate consumului uman,
respectiv furajrii animalelor (4). Reglementrile aprute n ultimii ani subliniaz
responsabilitatea productorilor i a furnizorilor n ceea ce privete participarea
lor la asigurarea calitii alimentelor (6).
Sistemul HACCP este un demers raional preventiv pentru a garanta
sigurana sanitar a alimentelor, prin supravegherea deosebit i inerea sub
control a acelor etape ale procesului tehnologic care sunt eseniale pentru
eliminarea contaminanilor periculoi pentru sntatea consumatorilor (1). n timp
ce programele tradiionale de supraveghere a siguranei alimentelor erau
concepute pentru a identifica problemele n produsul finit, HACCP este
implementat pentru a identifica riscurile poteniale i a garanta inerea lor sub
control n timpul procesului de producere sau de transformare a alimentelor (2).
Pentru carnea de porc, cele mai multe cauze raportate ale mbolnvirilor
datorate alimentelor obinute pe baza acesteia sunt de origine microbiologic (5).
Microbii sunt prezeni pretutindeni i pot intra n lanul alimentar n orice punct al
acestuia.

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Lucrarea de fa are ca obiectiv s prezinte eficacitatea implementrii


sistemului HACCP ntr-un abator de porci, pentru a garanta a sigurana sanitar a
crnii, evalund riscul contaminrii microbiologice.

MATERIAL I METODA

Studiul s-a desfurat ntr-un abator de dimensiuni medii situat n regiunea


de Nord-Est a Romniei, n trei etape, din luna noiembrie 2005 i pn n iulie
2006. Conducerea ntreprinderii a decis efectuarea acestui demers, att pentru a
respecta cerinele legislative (6, 7), ct i din dorina de a garanta clienilor o
carne de calitate i sigur din punct de vedere sanitar.
n prima etap, dup realizarea unei diagrame de producie, preciznd
manipulrile necesare fiecrei etape a fluxului tehnologic, timp de trei sptmni
a fost evaluat nivelul de igien al muncitorilor, curenia spaiilor de producie i
condiiile de lucru din ntreg abatorul. Au fost efectuate prelevri sanitare
sptmnale din numeroase puncte de lucru pentru a stabili gradul de
contaminare bacteriologic, cuprinznd cinci markeri: flora total mezofil,
Escherichia coli, Staphylococcus aureus, Clostridium perfringens, Salmonella sp.
n a doua etap s-a analizat i s-a conceput un sistem bazat pe principiile
HACCP, n dousprezece etape, conform recomandrilor lui Codex alimentarius
(3, 5). Pentru implementarea sistemului s-a dispus realizarea de investiii
materiale, prin schimbarea unor mijloace de producie dar i instruiri ale
personalului, mai ales formare pe tema bunelor practici de producie i a
respectrii regulilor de igien personal i tehnologic.
Dup analiza pericolelor i stabilirea riscurilor, echipa desemnat de
conducerea ntreprinderii pentru realizarea planului HACCP, a identificat etapele
din fluxul tehnologic n care aceste riscuri pot fi inute sub control. Stabilirea
punctelor critice de control a fost realizat prin schema de ntrebri succesive ale
arborelui de decizie (3). Fiecrui punct critic de control i s-au stabilit anumii
parametrii tehnologici, limite critice, care permiteau supravegherea precum i un
sistem de monitorizare i de nregistrare a lor.
Fiecrui post de lucru i mai ales celor considerate ca fiind critice pentru
inerea sub control a siguranei sanitare a alimentelor, au fost concepute
instruciuni de lucru i proceduri care au fost discutate cu cei implicai. Au fost
desemnate responsabiliti clare att pentru aciunile de monitorizare ct i pentru
activitile preventive, corective i de verificare.
n ultima etap a studiului s-a controlat stabilitatea sistemului prin
supravegherea ntregului flux tehnologic. Pentru verificarea eficienei din punct de
vedere bacteriologic, timp de trei sptmni au fost efectuate prelevri,n regim
matinal, de pe suprafee ale ustensilelor i ale carcaselor, prin metoda clasic a
testelor de sanitaie, care au fost trimise aceluiai laborator de bacteriologie,
acreditat, cu respectarea cerinelor referitoare la evitarea contaminrii. Toate
analizele bacteriologice au fost iniiate n maxim o or din momentul sosirii n
laborator, pentru aceiai cinci markeri din prima etap.
Analizele bacteriologice au utilizat metodele normalizate, oficiale.
Rezultatele au fost exprimate n uniti formatoare de colonii (ufc) pentru unitatea

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de suprafa. Interpretarea global a analizelor a inut cont de nivelul valorilor


admisibile, aprobate de reglementrile legale n vigoare.

REZULTATE I DISCUII

Prima etap, caracterizarea stadiului de igien a personalului i al spaiului


de producie, a prezentat numeroase neajunsuri, mai ales n cazul echipamentelor
care erau vechi, improprii sectorului agro-alimentar, mai ales conveierul i pereii,
tavanul i pardoseala halelor. Toate eantioanele prelevate au prezentat o depire
a nivelului acceptabil pentru flora mezofil total la 300C. Au fost identificate
microorganisme potenial patogene, mai ales Escherichia coli i Salmonella spp.
Investiiile materiale au constat din renovarea halelor de producie, a
sistemelor de evacuare a efluenilor i de ventilaie, readaptarea fluxurilor de
materiale i de personal pentru evitarea contaminrilor ncruciate i separarea
zonelor murdare de cele curate ale abatorului. A fost achiziionat o nou
camer frigorific cu sisteme de monitorizare a temperaturii, umiditii i
curenilor de aer. La fiecare post de lucru au fost instalate sterilizatoare pentru
cuite, cu ap fierbinte peste 830C, i o chiuvet pentru igiena minilor, dotat cu
robinet acionat cu genunchiul. Personalul a primit un echipament adecvat precum
i o instruire corespunztoare.
Dup punerea la punct a acestor prealabile de infrastructur i operaionale,
s-a trecut la conceperea sistemului HACCP de sigurana alimentar. S-a pornit de
la caracterizarea produsului cu evidenierea tuturor caracteristicilor importante
pentru sntatea consumatorului. Au fost evideniate constrngerile legale i
contractuale ale ntreprinderii, precum i destinaia prevzut pentru carcasele de
carne. De asemenea au fost evideniate caracteristicile materiilor prime porcii
destinai pentru sacrificare i materialele care vin n contact cu alimentul, mai ales
apa potabil. S-a constatat c la nivelul fermelor nu exist tehnici de gestiune care
s permit reducerea populaiilor de ageni patogeni; doar o bun igien i
respectarea biosecuritii la nivelul fermei precum i pstrarea strii de sntate a
animaleor sunt exigenele inconturnabile pentru reducerea populaiilor de
microorganisme.
Principalele manipulri care se efectueaz n abator, importante pentru
garantarea siguranei crnii i organelor interne, au fost descrise, insistndu-se
asupra celor care ar permite inerea sub control a anumitor pericole n etape ale
fluxului tehnologic. Analiza riscurilor asociate abatajului de porcine a fost
realizat innd cont de gradul de periculozitate al acestora dar i de probabilitatea
lor de realizare. Au fost identificate ase puncte critice (CCP) n care se pot ine
sub control riscurile biologice, chimice i fizice: recepia porcilor, splarea de
dup oprire i prlire, eviscerarea, controlul veterinar al crnii i organelor
interne, toaletarea carcaselor i rcirea crnii i organelor.

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Pentru fiecare punct critic de control au fost stabilite elemente de


monitorizare i limite critice pentru a garanta inerea sub control a riscurilor. Au
fost precizate i aciunile corective necesare a fi luate n cazul depirii limitelor
critice. Procedurile i instruciunile de lucru au fost documentate iar nregistrrile
arhivate pentru a demonstra viabilitatea sistemului iar verificarea funcionrii sale
a fost realizat periodic.
Dup implementarea sistemului HACCP s-a realizat cea de-a treia etap a
acestui studiu, respectiv verificarea funcionrii acestuia, insistndu-se pe analiza
microbiologic a carcaselor i a suprafeelor de lucru. Igiena mediului de lucru a
fost considerat corect n toate etapele de fabricaie, la fel ca i starea de igien a
materialului utilizat (cuite, crlige). Carcasele de porc obinute au avut o
ncrctur microbian mai redus comparativ cu cea descris nainte de
implementarea sistemului HACCP, fr s fie semnificativ statistic. Constatarea
unei reduceri a nivelului de ncrctur bacterian la nivelul suprafeelor
carcaselor a confirmat datele literaturii de specialitate (5).
Implementarea sistemului HACCP a fost corect perceput i total acceptat
de ctre angajaii abatorului, participarea i implicarea activ a acestora permind
buna desfurare a produciei n condiiile sanitare nou create, fiind vorba i de un
produs alimentar supus unor norme reglementare stricte (6, 7).

CONCLUZII

Analiza rezultatelor obinute dup implementarea unui sistem de siguran


a alimentelor bazat pe principiile HACCP ntr-un abator de porci demonstreaz c
garantarea producerii de carne i de organe interne de porc sigure din punct de
vedere sanitar se realizeaz doar prin implicarea activ a productorilor i a
transformatorilor, de la ferme i din abator. Riscurile microbiologice amintite n
acest articol trebuie s motiveze autoritile pentru a ntri supravegherea sanitar
a unor astfel de ntreprinderi, conform reglementrilor actuale, n care
autocontrolul este principiul esenial. Educaia sanitar a personalului i
promovarea igienei personale sunt componente de baz pentru implementarea cu
succes a principiilor analizei i inerii sub control a riscurilor sanitare.
Igiena alimentar din abatoarele de porci trebuie considerat de manier
integrat n programul general de supraveghere sanitar a zonei n care se afl
ntreprinderea (5), considernd-o ca o raportare fa de furnizarea apei potabile,
de tratarea apelor uzate, de ntreinerea mediului nconjurtor etc., probleme care
merit toat atenia pentru a garanta populaiei i mediului o stare de sntate
durabil.

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BIBLIOGRAFIE

1. Apostu S., 2004 Managementul calitii alimentelor. Sisteme moderne pentru


asigurarea calitii alimentelor, Editura Risoprint, Cluj-Napoca, pp.16-17.
2. Brdan Gh., 2006 Sigurana alimentelor, lucrri practice, FMV Iai, sept.- dec.
3. Codex alimentarius - Systme d'analyse des risques - Points critiques pour leur
matrise (HACCP) et Directives concernant son application (Appendice)
4. Comisia european - sigurana alimentelor: http://europa.eu.int/comm/food/
index_fr.htm
5. Generic HACCP Model for Pork Slaughter http://www.fsis.usda.gov/OPPDE/
nis/outreach/models/HACCP-14.pdf.
6. Rglement (CE) N 852/2004 du Parlement Europen et du Conseil du 29 avril 2004
relatif l'hygine des denres alimentaires.
7. Rglement (CE) 853 du Parlement Europen et du Conseil du 29 avril 2004 fixant
des rgles spcifiques dhygine applicables aux denres alimentaires dorigine
animale.

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EVALUAREA I CONTROLUL RISCURILOR ASOCIATE


PROCESRII CRNII PENTRU IMPLEMENTAREA
SISTEMULUI HACCP
CONTROL AND EVALUATION OF RISKS ASSOCIATED WITH MEAT PROCESSING
IN THE CONTEXT OF HACCP SYSTEM IMPLEMENTATION

L. ILIE, C. SAVU, Carmen PETCU


U.S.A.M.V. Bucureti

The industrial meat processing leads to the decrease of vitamin,


macro elements and trace elements content, which are followed by a
significant lower intake and the appearance of nutritional diseases.
Likewise, due to a prolonged technological processing of meat, there can be
noticed a decrease of certain mineral levels caused by the excessive use of
polyphosphates, accompanied by a visible increase of other mineral levels.
The evaluation of the risks associated with prime materials and ingredients
fabrication, processing, handeling, storing, distribution, dishes preparation
and consumption is considered the first step in HACCP (Hazard Analysis
and Critical Control Points) evaluation procedure. This system aims at
identifying and monitoring certain critical control points of the
technological process.

Key words: quality, risk, processing, quality maintenance.

Specia uman, pe parcursul existenei sale i-a dezvoltat un metabolism i


un echipament enzimatic complex care i permit s digere alimente procesate
termic, alimente care pentru diferite specii de animale omnivore sau carnivore
sunt greu de utilizat. Prelucrarea alimentului a determinat o serie de modificri
structurale ale acestuia (prelucrarea alimentului duce la distrugerea compoziiei
materiilor prime sau chiar la dispariia unor componeni), iar prelucrarea
industrial a determinat reducerea la minimum a celulozei i a fibrelor alimentare
din structura alimentelor, compui considerai iniial indezirabili i lipsii de
valoare trofic, ns lipsa acestora a determinat n timp apariia tulburrilor
intestinale i a altor afeciuni (constipaii cronice, diskinezii biliare,
hipercolesterolemii, afeciuni cardiovasculare etc.)
Procesarea industrial a crnii duce la scderea coninutului n vitamine,
macroelemente i microelemente, situaie care determin reducerea aportului i
apariia unor afeciuni careniale. De asemenea, datorit tehnologizrii crnii se
constat un nivel sczut de anumite sruri minerale ca urmare a folosirii n exces a
polifosfailor i creterea altor categorii de sruri minerale. Fosfatul tricalcic n
exces determin scderea absorbiei fosforului la nivel intestinal i totodat reduce
capacitatea metabolic de utilizare a acestuia la nivel organic.

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Evaluarea riscurilor asociate cu obinerea materiilor prime i a


ingredientelor, prelucrarea, manipularea, depozitarea, distribuia, prepararea
culinar i consumul produselor alimentare reprezint primul pas n
implementarea sistemului HACCP (Hazard Analysis and Critical Control Points),
sistem care i propune s identifice i monitorizeze anumite puncte critice de pe
traseul procesului tehnologic. inerea sub control a parametrilor care
caracterizeaz aceste puncte critice i msurile corective aplicate, garanteaz
securitatea alimentelor obinute.

MATERIAL I METOD

Evaluarea riscurilor, primul principiu al sistemului HACCP, vizeaz


identificarea pericolelor i modului n care acestea pot fi reduse sau eliminate.
Analiza riscurilor este asociat cu monitorizarea aciunilor de obinere i recoltare
a materiilor prime i auxiliare, de prelucrare, manipulare, depozitare i distribuie,
inclusiv prepararea culinar i consumul produselor alimentare.
Un potenial risc este considerat un agent biologic, fizic sau chimic, care
prin proprietile sale poate compromite sigurana unui aliment.
Evaluarea riscurilor se realizeaz n dou etape:
- evaluarea tipului de produs n funcie de riscurile asociate acestuia;
- evaluarea riscurilor n funcie de severitate.
Includerea produsului ntr-o anumit categorie de periculozitate se face pe
baza urmtoarelor caracteristici:
- produsul conine sau nu ingrediente sensibile;
- fluxul tehnologic presupune sau nu o etap n care este posibil
distrugerea eficient a microorganismelor periculoase sau a celorlalte
riscuri identificate;
- exist un risc major de contaminare a produsului dup ncheierea
procesului de fabricaie;
- exist pericolul unei manipulri necorespunztoare n timpul
transportului, vnzrii i pregtirii culinare care s fac produsul
periculos pentru consum;
- dup ambalare, produsul necesit tratamente termice sau pregtire
culinar.

REZULTATE I DISCUII

Analiza riscului: activitatea de colectare i evaluare a informaiilor privind


riscurile i condiiile care duc la apariia lor pentru a decide care dintre acestea
sunt relevante pentru sigurana alimentului i care vor fi abordate n planul
HACCP.
Pericolele biologice sunt clasificate n urmtoarele categorii: bacteriene,
virale sau parazitologice. Majoritatea produselor alimentare sunt un substrat ideal
pentru dezvoltarea microorganismelor, n special atunci cnd acestea sunt pstrate
perioade ndelungate la temperatura mediului ambiant.
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Principalul obiectiv va fi aplicarea de msuri preventive n scopul


distrugerii, eliminrii sau reducerii riscurilor, prevenirii recontaminrii i inhibrii
dezvoltrii microorganismelor precum i a producerii de toxine.
Microorganismele pot fi distruse prin tratament termic (pasteurizare sau
sterilizare) sau inhibare prin refrigerare, congelare, uscare, adugarea unor
conservani. Dup eliminarea microorganismelor, trebuie luate msuri pentru
prevenirea recontaminrii. n situaia n care microorganismele nu pot fi total
eliminate din produsele alimentare, vor fi luate msuri de inhibare a dezvoltrii
microorganismelor remanente i a producerii de toxine. Dezvoltarea
microorganismelor poate fi inhibat prin caracteristicile intrinseci ale produselor,
prin adugarea de sare sau ali conservani sau alegerea unor metode de ambalare
i temperaturi de depozitare corespunztoare.
Pericole chimice: dei se cunoate faptul c toate substanele chimice pot fi
toxice la o anumit concentraie, producia modern de alimente nu poate fi
conceput fr utilizarea substanelor chimice. n tabelul 1 este prezentat
clasificarea substanelor chimice prezente n produsele alimentare, care pot
constitui un risc la adresa sntii consumatorilor.
Tabelul 1
Categorii de risc chimic

Substane chimice Micotoxine (ex. aflatoxine)


naturale Scombrotoxina (histamina)
Ciguatoxina
Toxine din ciuperci
Toxine din crustacei (toxina crustaceica paralitica, diareica,
neurotoxica, amnezica)
Alcaloizi pirolizidinici
Fitohemaglutinine
Fenoli policlorurati
Substane chimice Substane chimice agricole
adugate Pesticide, fungicide, ngrminte, insecticide,
antibiotice, hormoni de cretere
Substane interzise
Metale toxice i combinaii ale acestora
Pb, Zn, As, Hg, cianuri
Aditivi alimentari
Direci conservani, poteniatori de arom,
aditivi nutriionali, colorani
Indireci substane provenite de la utilaje :
lubrifiani, ageni de curire i
dezinfecie, substane de acoperire,
vopseluri
Substane chimice adugate intenionat (sabotaje)

n funcie de doza care a fost ingerat, aceste substane chimice pot


determina mbolnvirea sau moartea imediat a consumatorului sau n timp, prin

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acumularea lor n organism. n tabelul 2 sunt prezentate cteva modaliti de


prevenire a apariiei riscurilor chimice la procesarea crnii.
Tabelul 2
Modaliti de meninere sub control a pericolelor chimice

Control nainte Precizarea compoziiei materiei prime;


de recepie Certificate de calitate emise de ctre furnizor;
Teste de recepie.
Control nainte Stabilirea scopului utilizrii substanelor chimice;
de utilizare Verificarea calitii substanelor utilizate;
Verificarea cantitii utilizate.
Controlul condiiilor de Meninerea substanelor chimice n condiii conforme cu
depozitare i manipulare precizrile din avizele de nsoire ale acestora.
Inventarierea Efectuarea unui inventar al substanelor chimice necesare,
substanelor chimice al dozelor i modului de utilizare a acestora.
existente

Pericole fizice: orice particul sau corp fizic ce nu se gsete n mod normal
n aliment i care poate provoca mbolnvirea sau rnirea unei persoane,
constituie un risc fizic. Acest lucru se explic nu att prin periculozitatea pe care o
prezint corpurile strine, ci mai ales prin faptul c ele pot fi uor detectabile i
observabile cu ochiul liber. Cele mai rspndite corpuri strine sunt prezentate n
tabelul 3.
Tabelul 3
Principalele pericole fizice

Efecte asupra
Material Surse
consumatorului
Ambalaje, corpuri de iluminat, ecrane ale
Sticla Tieturi, sngerare
aparatelor de msur
Lemn Tieturi, nepturi Terenuri, palei, cutii, cldiri
Pietre Rniri Terenuri, cldiri
Achii metalice Tieturi, infecii Utilaje, terenuri, cabluri, lucrtori
Insecte mbolnviri, traume Terenuri, mediul de lucru
Oase Rniri, traume Terenuri, prelucrare neglijent
Plastic Rniri, tieturi, infecii Terenuri, ambalaje, palei, angajai
Efecte Rniri, tieturi, spargerea
Angajai
personale dinilor

n general, prin practici bune de lucru aceste riscuri fizice pot fi prevenite
cu uurin.
Monitorizarea este o secven planificat de observaii i msurtori,
realizate pentru a putea aprecia dac un punct critic de control este sub control i
se finalizeaz printr-o nregistrare exact, utilizat ulterior n procesul de
verificare.

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Monitorizarea este esenial n managementul siguranei alimentelor. Dac


monitorizarea indic o tendin de pierdere a controlului n punctele critice ale
procesului de fabricaie, atunci se poate aciona pentru readucerea procesului sub
control nainte de apariia unor abateri de la inocuitate.
Monitorizarea este utilizat pentru a se determina momentul n care s-a
pierdut controlul i au aprut abateri ntr-un punct critic de control i n acest
moment, se vor aplica aciunile corective.
Monitorizarea trebuie s fie capabil s detecteze orice pierdere a
controlului i s furnizeze informaii n timp util, astfel nct s poat fi aplicate
msuri corective, evitnd sau reducnd la minimum riscul producerii unor
produse neconforme.
Monitorizarea poate fi realizat prin msurtori efectuate asupra unor
eantioane prelevate conform unui plan de eantionare realizat pe baze statistice
sau pe baza cerinelor legale i de reglementare.
Exist cinci modaliti principale de monitorizare a unui punct critic de
control, i anume: observarea vizual, aprecierea senzorial, msurtori fizice,
testri chimice i analize microbiologice.
Pstrarea nregistrrilor este o parte integrant a monitorizrii i va
demonstra c au fost urmate ntocmai procedurile de-a lungul procesului
tehnologic. Existena acestor nregistrri ofer produsului i procesului o
caracteristic foarte important, trasabilitatea.

CONCLUZII

1. Specificitatea sistemului HACCP este dat de faptul c, n vreme ce


nivelul calitativ al produselor fabricate poate fi la libera alegere a productorului
sau poate face obiectul negocierii, inocuitatea produselor este un element
obligatoriu.
2. Beneficiile acestui sistem sunt: sigurana produselor alimentare fabricate,
utilizarea mai bun a resurselor i un rspuns mai rapid la problemele aprute n
producie.
3. Este de importan vital, pentru societate i pentru consumatori,
cooperarea ntre industrie, comer, organizaii ale consumatorilor i autoritile
responsabile. Programele de instruire comun pot ncuraja i menine un dialog
continuu ntre prile menionate i pot crea un climat de nelegere n aplicarea
practic a sistemului HACCP.

BIBLIOGRAFIE

1. Barzoi D., Apostu S., 2002 - Microbiologia produselor alimentare, Editura Risoprint,
Cluj-Napoca.
2. Carol W., Sara Mortimore, 2001 - The HACCP Training Resource Pack, Publisher
Springer.

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3. Deniss, R.A., 2002 - Quality Audits for Improved Performance, ASQ Quality Press.
4. Kevin, M.Q., 2005 - A good guide to the HACCP Auditor Test, Illinois United States.
5. Mitrea, I.S., Carmen Petcu, Savu, Gh., 2003 Sigurana alimentelor prin aplicarea
sistemului HACCP, Editura Bogdana, Bucureti.
6. Paster, T., 2006 - HACCP Food Safety Employee Manual, Publisher Wiley, John &
Sons.
7. Russell, J.P., 1999 - The Quality Audit Handbook, ASQ Quality Press.
8. ***, 2001 - American Society for Quality - The Certified Quality Auditor's Haccp
Handbook: Asq Food, Drug, and Cosmetic Division, ASQ Quality Press, 2001.

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OBSERVAII PRIVIND MICROFLORA CRNII


DE PASRE PROASPETE I REFRIGERATE
OBSERVATION CONCERNING FRESH
AND REFRIGERATED POULTRY MICROFLORA

Elena Narcisa NECULI1, V. FLORITEAN2,


Elena IAN3, M. CARP-CRARE2
1
D.S.V.S.A. Vaslui
2
U.S.A.M.V. Iasi
3
L.S.V.S Iai

A study was carry out in order to examine the microflora of poultry


carcasses before, during and after refrigeration storage on some common
spoilage bacteria in the native microflora of broiler carcasses. Chilled
carcasses, obtained from a commercial processing facility, were examined
prior, during and after 7 days of storage at 4C. The total aerobic plate
cont, coliforms, E. coli and Pseudomonas spp. in the bacterial flora were
enumerated.
The size of the population of spoilage bacteria on processed
carcasses stored at 4 degrees C for 7days was determined. The total
aerobic plate count /g, about log 3.5 on the beginning of the storage,
increased by cca. 2.5 log after 7 day. Coliforms and Escherichia coli
counts/g slightly decrease over this period of storage. The increase in the
population of bacteria on the carcasses was the effect of phychrotrophic
bacteria, Pseudomonas spp. being bacteria that have revealed the most
important increasing values during the refrigeration storage.

Key words: Poultry meat, Meat refrigeration, Meat storage

Refrigerarea este la ora actual una dintre cele mai folosite metode de
conservare a produselor animale de origine animal i n special a crnii, ntruct
ofer consumatorului carne cu caracteristici organoleptice i nutritive foarte
apropiate de cele ale crnii proaspete. Durata de conservare a crnii de pasre prin
refrigerare este limitat n principal de microorganismele care contamineaz
carcasa. Carnea de pasre, imediat dup prelucrarea n abator, are o ncrctur
bacterian mic de ordinul a 3 - 4 log ufc/cm2 care ajunge, uneori, n reeaua de
comercializare la 5 i chiar 7 log ufc/cm2, mai ales n cazul cnd intervin abuzuri
ce expun carnea la aciunea nefavorabil a unor factori ce stimuleaz dezvoltarea
bacterian. (2, 3, 4, 7, 9, 8, 10, 14)
n plus microflora crnii de pasre refrigerat poate fi reprezentat uneori
de microorganisme patogene (Salmonella spp., Listeria monocytogenes, Yersinia
enterocolitica, Aeromonas hydrophila, Bacillus cereus etc.) fapt care constituie un
risc potenial pentru consumatori. (6, 8, 12, 13)

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Prezenta lucrare are drept scop monitorizarea dinamicii microflorei de la


suprafaa i din profunzimea crnii de pasre pstrat la temperatura de 4C,
temperatur caracteristic pentru pstrarea crnii de pasre n reeaua de
desfacere.

MATERIAL I METODA

Cercetrile au fost efectuate n perioada iulie - octombrie 2005 pe un


numr de 25 carcase de pasare provenite de la o unitate de abatorizare a
psrilor din judeul Vaslui. De la carcase, imediat dup rcire, sau excizat
aseptic probe de esut muscular cu si fr piele din regiunea pieptului de
aproximativ 200g. Probele au fost ambalate aseptic i transportate la laborator n
condiii corespunztoare pentru a se preveni modificarea caracteristicilor
microbiologice ale acestora.
In laborator fiecare prob a fost mprit n 8 porii mai mici (aprox. 25g)
care au fost puse n plci Petri sterile i pstrate la termostat la 4C timp de 7 zile.
Probele au fost examinate bacteriologic la scurt timp dup recoltare i apoi
zilnic, pe toat perioada pstrrii la temperatura de refrigerare. S-a urmrit
numrul total de germeni aerobi mezofili, bacteriile coliforme, E.coli i bacteriile
din genul Pseudomonas.
Prima diluie din care s-au preparat ulterior diluiile necesare examenului
bacteriologic s-a obinut din 15 g esut muscular amestecat cu 135 ml apa
peptonat steril n pungi speciale sterile i omogenizare la Stomacher timp de 5
minute.
Determinarea NTG s-a efectuat pentru diluiile 10-2, 10-3 i 10-4, pe agar
nutritiv, prin nglobarea a 0,1 ml inocul. Pentru fiecare diluie s-au efectuat cte
dou plci. Plcile nsmnate au fost incubate la 37C timp de 24 - 48 ore.
Pentru determinarea bacteriilor coliforme s-a utilizat metoda ISO 4832.
n cazul ncrcturilor mici de coliformi (< de 2 log/ml) determinarea s-a
efectuat prin diluii zecimale, din fiecare diluie introducndu-se cte 1 ml n cte 3
eprubete cu mediu BBLV (bulion lactozat cu sruri biliare i verde briliant) i tub
Durham. S-a incubat 24 48 ore la 37 C. Interpretarea s-a fcut dup producia
de gaz i calcularea mediei de 3 cifre (funcie de nota primit de fiecare din cele 3
eprubete cu mediul BBLV), media obinut fiind interpretat cu ajutorul tabelului
Mac Crady.
Pentru identificarea bacteriei Escherichia coli s-a utilizat metoda ISO 4832
iar pentru determinarea numrului probabil de E. coli s-a utilizat metoda SR ISO
7251.
Identificarea i numrarea bacteriilor din genul Pseudomonas s-a efectuat
din diluiile seriate conform ISO 13720 folosind mediul CFC (cephaloridine-
fucidin-cetrimide) cu incubare la 25C timp de 48 ore.

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REZULTATE I DISCUII

Studiile efectuate, pn n prezent, pentru a evalua microflora crnii de


pasre proaspt i refrigerat au demonstrat marea diversitate a acesteia. Mai
mult de 25 de genuri de microorganisme au fost evideniate ca fcnd parte din
microflora crnii de pasre proaspete, majoritatea acestora fiind microorganisme
de poluare dar i microorganisme patogene. (2, 7, 8, 10, 11, 13)
Numrul total de germeni prezeni pe carcasele examinate a avut valori de
3,53 0,12 (log UFC/g eroarea standard) la scurt timp de la recoltarea probelor,
valori asemntoare cu cele publicate de ali autori, dar care este mare dac inem
cont c am efectuat determinrile preponderent pe esut muscular (6, 7, 10).
Numrul total de germeni a crescut progresiv pe toat perioada de
meninere la temperatura de 4C atingnd valori de 6,2 0,21 log ufc/g n cea de-
a aptea zi, moment n care am constatat i uoare modificri ale caracterelor
organoleptice, carnea avnd suprafaa uor lipicioas i un miros slab acid.
Coliformii sunt microorganisme Gram-negative, nesporulate, care
fermenteaz lactoza i produc colonii negricioase cu luciu metalic pe mediul
Endo. Coliformii sunt microorganisme ubicvitare care se folosesc, nc, n
industria alimentar ca microorganisme indicator a strii de igien a fluxului
tehnologic al alimentului. (2, 3, 8)
n studiul efectuat coliformii au avut, iniial valoarea de 2,73 0,35, iar pe
parcursul refrigerrii au sczut progresiv, lent, ajungnd la finalul celor apte zile
la 2,46 0,44. Evoluia coliformilor nu este surprinztoare deoarece acetia sunt,
n marea lor majoritate, microorganisme mezofile, creterea lor la temperaturi
sczute fiind disgonic. (2, 3, 8)
Evoluia numrului de E.coli a fost intens descresctoare fiind parametrul
cel mai afectat de temperatura sczut. Astfel, dac iniial numrul de E. coli/g a
fost de 2,12 0,11, reprezentnd cca. 78% din totalul coliformilor, la sfritul
perioadei de pstrare a ajuns la 1,16 0,05 fiind aprox. 47% din numrul
coliformilor. Dei marea majoritate a tulpinilor de E. coli izolate de la psri sunt
inofensive, n cadrul speciei exist i tulpini cu patogenitate ridicat pentru om.
Bacteriile din genul Pseudomonas, alturi de alte genuri (Acinetobacter,
Moraxella, Enterococcus, Alcaligenes. etc), fac parte din categoria
microorganismelor psichrotrofe i formeaz microflora predominant a
produselor refrigerate. (1, 2, 3, 8)
Aa cum era de ateptat, dei numrul iniial de Pseudomonas spp. a fost
mai mic de 1 log ufc/g, acestea s-au dezvoltat bine la temperatura de 4C
ajungnd la finalul testului la 3.13 0,42 ceea ce reprezint aprox. 50% din NTG.
(Tabelul 1)
Dup cum se poate observa, creterea NTG s-a realizat n special prin
aportul microorganismelor psichrotrofe i mai puin a altor microoganisme.
(Fig. 1)

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Datele din literatur susin c la temperatura de 4-5C carnea de pasre i


pstreaz nealterate caracterele organoleptice pn la 9 zile, perioad ce poate fi
mrit prin reducerea temperaturii de pstrare i ambalarea carcaselor.
Carnea de pasre devine inapt pentru consum datorit modificrilor
organoleptice atunci cnd numrul total de germeni de la suprafaa crnii atinge
valori mai mari de 7 log ufc/cm2.
n condiiile studiului efectuat carnea s-a depreciat rapid datorit
ncrcturii microbiene iniiale mari. Bacteriile psichrotrofe din genul
Pseudomonas la care se adaug i unii coliformi din genul Enterobacter, se pot
dezvolta la temperaturi sczute i prin metabolismul diversificat care induc
degradarea crnii de pasre.
Tabelul 1
Evoluia parametrilor microbiologici in carnea de pasre la temperatura de 4C

Durata de pstrare la 4C (zile)


Parametrul
0 1 2 3 4 5 6 7
NTG 3,51 3,61 3,84 4,27 4,35 4,66 5,48 6,22
Coliformi 2,73 2,75 2,78 2,64 2,61 2,53 2,52 2,46
Escherichia coli 2,14 2,22 2,15 2,11 1,83 1,55 1,35 1,16
Pseudomonas spp. <1 1,22 1,48 1,92 2,33 2,64 2,91 3,10

5
Log CFU/g

0
-1 0 1
NTG 2 3 4 5 6
Coliformi 7
Escherichia coli Pseudomonas spp.

Fig. 1 Dinamica creterii microorganismelor din carnea de pasre la temperatura de 4C

CONCLUZII

Microorganismele care formeaz microflora normala a crnii de pasre se


pot dezvolta la temperatura de refrigerare conducnd la degradarea crnii de
pasre ntr-un timp relativ scurt;

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Microorganismele din specii psichrotrofe (genul Pseudomonas) se gsesc


frecvent i n numr suficient de mare pentru a induce alterarea precoce a crnii
de pasre conservate prin refrigerare;
Un rol important n prelungirea perioadei de conservabilitate a crnii de
pasre l are ncrctura microbian iniial, ca urmare este necesar respectarea
tuturor msurilor de reducere a numrului de microorganisme prin folosirea unor
tehnologii de prelucrare a psrilor n abatoare corespunztoare i prin respectarea
msurilor de igien pe tot parcursul fluxului tehnologic;
Se impune rcirea imediat a carcaselor de pasre, ambalarea acestora i
respectarea temperaturii de refrigerare 2-4C pe tot fluxul, de la abator pn la
consumator;
n condiiile actuale este necesar reducerea timpului de pstrare la
temperatura de refrigerare la 3 zile fapt care ar asigura calitatea crnii de pasre i
ar reduce posibilitatea apariiei de neplceri consumatorilor

BIBLIOGRAFIE

1. Ayres, J.C., 1960 - The Relationship of Organisms of the Genus Pseudomonas as


the Spoilage of Meat, Poultry and Eggs. Journal of Applied Bacteriology 23, 471-
486
2. Barnes, E.M., Thornley, M.J., 1966 - The spoilage flora of eviscerated chickens
stored at different temperatures. Journal of Food Technology 1:113-119
3. Brzoi D., 1986 - Microbiologia produselor alimentare de origine animal Editura
Ceres, Bucureti
4. Brzoi D., Apostu S., 2002 - Microbiologia produselor alimentare Ed. Risoprint, Cluj-
Napoca
5. Blood, R.M., Jarvis, B., 1974 - Chilling of Poultry: The Effects of Process
Parameters on the Level of Bacteria in Spin-Chiller Waters Journal of Food
Technology 9, 157-169
6. Cox, N. A. (1990) "The Presence of Yersinia enterocolitica and Other Yersinia
Species on the Carcasses of Market Broilers" Poultry Science 69, 482-485
7. Elliott, R.P., Michener, H.D., 1961 - Microbiological Standards and Handling Codes
for Chilled and Frozen Foods. A Review Applied Microbiology 9, 452-468
8. Jay, J.M., 2000 - Fresh meat and poultry in Modern Food Microbiology sixth ed.
Aspen Publishers, Inc. Gaithersburg, Maryland, 59-87
9. Kraft, A.A., 1971 - Microbiology of Poultry Products Journal of Milk and Food
Technology 34, 23-29
10. May, K.N., Irby, J.D., Carmon, J.L., 1961 - Shelf life and bacterial counts of excised
poultry tissue. Food Technology 16, 66-68
11. McMeekin, T.A., 1975 - Spoilage association of chicken breast muscle. Applied
Microbiology 29, 44-47
12. Sum, A.R. Editor, 2001 - Poultry Meat Processing CRC Press, 121-135
13. Sindilar E., Greceanu Al., Carp-Carare M., Elena Verdes, 1992-1993 - Investigatii
experimentale privind modificarile produse de microflora psihrotrofa pe carnea
refrigerata si congelata de pasare. Lucrari stiintifice, vol. 35-36, 416-420
14. Waldroup, A.L., 1996 - Contamination of Raw Poultry with Pathogens, World Poultry
Science Journal 52, 7-25

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INFLUENA CONDIIILOR DE CRETERE


I A TRANSPORTULUI ASUPRA CALITII
MICROBIOLOGICE ALE CRNII DE PASRE
INFLUENCE OF WELFARE AND TRANSPORTATION CONDITIONS
ON MICROBIOLOGICAL QUALITY OF POULTRY MEAT

Elena Narcisa NECULI


D.S.V.S.A. Vaslui
V. FLORITEAN, M. CARP-CRARE
U.S.A.M.V. Iai

A study was conducted to determine effects of welfare, and


transportation conditions on poultry carcasses microflora. Samples were
taken from farm, from transportation vehicles and from chickens before
transportation, prior to abattoir processing and from carcasses short time
after defeathering. The aerobic plate count, coliforms and Escherichia coli
were estimate. Under the conditions of this experiment, it appears that
welfare and transportation condition do affect the level of contamination on
the exterior of birds entering the processing facility. The flora identified
was mainly the same and the level of contamination of carcasses was
positive correlate with farm microflora and stress transportation of
chickens.

Key words: Poultry meat, Poultry quality, Transportation, Welfare

Calitatea crnii de pasre este afectat de numeroi factori ce acioneaz


att nainte ct i dup sacrificare. Factorii care acioneaz antemortem pot fi
factori cu efect pe termen lung, pentru c acioneaz pe parcursul ntregii viei
economice a psrilor (factori genetici, fiziologici, nutriionali, patologici, etc) i
factori cu aciune de scurt durat care acioneaz cu preponderen pe parcursul
ultimelor 24 de ore de via a psrilor (condiii de recoltare, de transport, dieta
alimentar, imobilizarea, asomarea, sngerarea, etc.). Aceti factori influeneaz
att greutatea carcasei i capacitatea metabolic a esutului muscular ct i
calitatea microbiologic a carcasei. (1, 2, 3, 6)
Aspectele care privesc factorii cu aciune de scurt durat, datorit
importanei lor, au captat atenia procesatorilor de carne de pasre ntruct prin
managementul corect al acestora se poate obine carne de pasre de calitate lipsit
de pericole pentru consumator. (2, 5, 4)
n acest studiu am urmrit modul n care unii factori de cretere din fermele
de psri i transportul psrilor de la fermele de cretere la abator, influeneaz
calitatea microbiologic a crnii precum i stabilirea unor msuri de corectare a
deficienelor care pot conduce la deprecierea calitii crnii de pasre.

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MATERIAL I METOD

Studiul a fost efectuat n perioada martie septembrie 2005 pe un numr


de 90 pui destinai sacrificrii n cadrul unei ferme de profil din judeul Vaslui i n
abatorul acesteia. Determinrile microbiologice au fost efectuate n laboratorul
uzinal al unitii de tiere i n Laboratorul Sanitar Veterinar Judeean Vaslui. Puii
au fost grupai n nou loturi n funcie de data atingerii greutii de sacrificare.
Puii au fost crescui cte trei loturi n hale separate n condiii de microclimat
diferite. Transportul s-a efectuat cu mijloace de transport autorizate prin gruparea
a cte trei loturi (cte unul din fiecare hal de cretere).
S-au monitorizat:
- microclimatul (temperatura i umiditatea) din halele de cretere pentru
o perioad de 10 zile anterior sacrificrii;
- microbiota din halele de cretere (probe din ap, furaje, aternut i
din aerul din adposturi), n ziua livrrii psrilor pentru sacrificare;
- durata perioadei de diet alimentar pentru fiecare lot n parte (timpul
scurs din momentul sistrii furajrii n ferma de cretere pn n
momentul sacrificrii);
- ncrctura microbian din mijlocul de transport nainte i dup
transportarea psrilor la abator,
- calitatea microbiologic a carcaselor provenite de la pui, dup
sngerare i dup oprire-deplumare.
Temperatura i umiditatea s-a msurat i nregistrat cu aparatur adecvat
de 4 ori pe zi, halele de cretere fiind dotate cu aparatur modern de
monitorizare a parametrilor de microclimat cu posbilitatea corectrii deficienelor.
Examenele microbiologice au constat n determinri ale urmtorilor
parametri: NTG, bacterii coliforme, E. coli i bacterii sulfito-reductoare.
Recoltarea probelor s-a efectuat cu respectarea procedurilor impuse prin
standarde specifice pentru fiecare determinare.
Pentru determinrile microbiologice s-au utilizat metode uzuale, impuse
prin stasuri interne i internaionale, dup cum urmeaz:
- Pentru determinarea N.T.G - SR ISO 6610/1997;
- Pentru bacterii coliforme - SR ISO 4832 i tehnica MPN cnd a fost
necesar;
- SR ISO 4832 i SR ISO 7251/1996 pentru stabilirea numrului de
Escherichia coli;
- Determinarea numrului de bacterii sulfito-reductoare s-a efectuat pe
mediul cu sulfit de sodiu i citrat de fier, topit i rcit la 45C n care s-
a nsmnat cte 1 ml din diluiile zecimale. S-a incubat n
anaerobioz la 37C timp de 24 48 ore, dup care au fost numrate
coloniile specifice.
Rezultatele au fost interpretate statistic, calculndu-se media pentru fiecare
lot i parametru determinat i coeficienii de corelaie caracteristici evoluiei
parametrilor n condiiile studiului.

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REZULTATE I DISCUII
n perioada studiului, microclimatul din halele de cretere s-a pstrat n
limite relativ constante. Temperatura, dei a nregistrat unele variaii, s-a meninut
n medie, la cele nou loturi (grupate cte trei in hale diferite) ntre 20,4 i 24,6C.
Umiditatea relativ a adposturilor, ca factor de confort, s-a meninut ntre 72 i
78% cu variaii relativ mari ntre zi i noapte. Cele nou loturi au fost supuse
naintea sacrificrii unei diete difereniate ca timp pentru a evidenia efectul
acesteia asupra calitii microbiologice a crnii. (Tabelul 1)
Tabelul 1
Variaia temperaturii i umiditii pe perioada analizat

Temperatura C Umiditatea relativ % Durata


Lot dietei
Minim maxim media minim maxim media (ore)
1 18,1 22,3 20,4 48 86 78 6
2 18,1 22,3 20,4 48 86 78 8
3 18,1 22,3 20,4 48 86 78 10
4 19,6 23,5 22,1 46 85 76 6
5 19,6 23,5 22,1 46 85 76 8
6 19,6 23,5 22,1 46 85 76 10
7 20,2 25,1 24,6 44 82 72 6
8 20,2 25,1 24,6 44 82 72 8
9 20,2 25,1 24,6 44 82 72 10

n ceea ce privete microbiota adposturilor valorile cele mai mari au fost


evideniate la nivelul pardoselii unde NTG a variat, n medie, ntre 5,9 i 7,5,
numrul bacteriilor coliforme, ntre 3,5 i 4,9 iar bacteriile sulfito-reductoare
ntre 3,1 i 3,4. Aerul i apa au fost mai puin contaminate, apa avnd un numr de
coliformi ntre 1,1 i 1,6 (log UFC/cm2, g sau m3). (Tabelul 2)
Tabelul 2
Microbiota halelor de cretere a puilor destinai sacrificrii
2 3
Media (log10 ufc/cm , m , ml)
Lot Pardoseal Aer Apa
B. B B.
NTG B colif. NTG NTG B colif. E.coli
sulfitored Colif. sulfitored
1-3 6,2 4,2 3,1 4,8 2,5 2,1 2,1 1,6 1,1
4-6 7,5 4,9 3,4 5,6 3,4 2,7 2,4 1,2 <1
7-9 5,9 3,5 3,3 4,3 2,1 2,6 2.3 1.1 <1

Mijloacele de transport i cutile cu care s-a efectuat transportul puilor la


abator, pregtite (curate i decontaminate), au avut valori mici ale parametrilor
microbiologici monitorizai, ns la determinrile fcute la abator, dup
descrcarea puilor, ncrctura microbian a crescut n medie cu aproximativ 2
log. (Tabelul 3)

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n tabelul 4 sunt prezentate valorile parametrilor microbiologici determinai


dup sngerarea puilor, pentru a se putea evalua evoluia acestora. Determinrile
au fost repetate i dup operaiunea de oprire deplumare, valorile obinute fiind
semnificativ mai mici.
Tabelul 3
Rezultatele examenului microbiologic al mijloacelor de transport

ncrctura microbian a mijloacelor de transport


Lot nainte de transport Dup transport
B. B. B. B.
NTG E. coli NTG E. coli
colif. sulfitored. colif. sulfitored.
1 3,5 2,2 1,4 2,1 5,9 4,7 3,3 2,7
2 4,1 2,8 1,5 2,3 6,1 4,8 3,7 2,5
3 5,2 2,4 1,1 3,1 6,3 4,6 3,4 -
4 3,5 2,2 1,4 2,1 5,9 4,7 3,3 2,7
5 4,1 2,8 1,5 2,3 6,1 4,8 3,7 2,5
6 5,2 2,4 1,1 3,1 6,3 4,6 3,4 -
7 3,5 2,2 1,4 2,1 5,9 4,7 3,3 2,7
8 4,1 2,8 1,5 2,3 6,1 4,8 3,7 2,5
9 5,2 2,4 1,1 3,1 6,3 4,6 3,4 -

Tabelul 4
Calitatea microbiologic a carcaselor de pasre n abator
2
Calitatea microbiologic a carcaselor (log10 ufc/cm )
Lot Dup sngerare Dup deplumare
B. B.
NTG B. colif. E. coli NTG B. colif. E. coli
sulfitored. sulfitored.
1 6,8 4,3 1,5 2,4 5,1 3,6 2,1 1,7
2 6,5 3,8 1,4 2,8 5,2 2,4 1,2 2,2
3 6,2 3,7 1,4 2,1 4,6 2,4 1,5 1,9
4 7,5 4,5 2,1 2,2 6,3 2,8 2,2 1,5
5 7,1 4,2 1,7 2,9 6,1 2,6 1,8 1,2
6 6,7 4,1 1,5 2,3 5,5 3,1 1,4 1,6
7 6,2 3,6 1,2 2,7 4,8 2,1 1,1 1,1
8 6,3 3,5 1,1 1,2 4,1 2,2 1,6 -
9 5,9 3,9 1,3 1,5 4,2 2,4 1,6 -

Interpretarea statistic a rezultatelor obinute, pe parcursul tuturor punctelor


enumerate, a relevat o corelaie pozitiv evident, n special, ntre valorile
obinute n urma determinrilor fcute pe carcase i valorile obinute la
determinrile efectuate n adposturi - pardoseal i aer (coeficient de corelaie
apropiat de valoarea unitar). Rezultatele microbiologice de la carcase s-au
corelat negativ dar mai puin evident cu variaiile temperaturii, cu durata dietei i
cu valorile rezultatelor obinute la mijloacele de transport. (Tabele 5 i 6)

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Relaia invers proporional cu durata dietei este explicabil prin faptul c


psrile care au fost supuse unei diete mai lungi au tubul digestiv golit iar
contaminarea pe timpul transportului este mult redus. Corelaie negativ cu
temperatura se explic prin faptul c psrile consum mai puin hran atunci
cnd temperatura este mai mare.
Rezultatele obinute ne permite s afirmm c sursa cea mai important de
contaminare cu microorganisme a carcaselor de pasre sunt psrile nsi. n
ceea ce privete umiditatea, corelaia evideniat nu este elocvent ntruct
variaiile constate pe parcursul studiului au fost prea mici.

CONCLUZII

Principalii factori cu aciune de scurt durat care afecteaz calitatea crnii


de pasre, aa cum se desprinde din rezultatele obinute, sunt cei legai de igiena
halelor de cretere a puilor destinai sacrificrii;
Durata dietei alimentare are un rol important n obinerea unor carcase cu
un grad redus de contaminare. Se recomand o dieta care s permit evacuarea
coninutului intestinal, dar care s nu afecteze greutatea psrilor sau rezistena
intestinelor la aciunea factorilor mecanici din cursul eviscerrii;
Temperatura i umiditatea halelor de cretere, ca factori cu aciune de
scurt durat, influeneaz indirect calitatea microbiologic a carcaselor prin
disconfortul pe care l creeaz psrilor;
Transportul puilor de la fermele de cretere la abator afecteaz calitatea
microbiologic a carcaselor, mijlocul de transport fiind locul unde contaminarea
ncruciat este frecvent, intensitatea acesteia fiind variabil;
n contextul actual, avnd n vedere importana calitii microbiologice a
crnii de pasre, se impune cu necesitate cunoaterea tuturor factorilor care pot
influena acest aspect i stabilirea unor programe de supraveghere i control
pentru un bun management al calitii crnii.

BIBLIOGRAFIE

1. Duke, G.E., Basha, M., Noll, S., 1997 - Optimum duration of feed and water removal
prior to processing in order to reduce the potential for fecal contamination in
turkeys, Poultry Science 76, 516
2. Hargis, B.M., Caldwell, D.J., Byrd, A.J., 2001 - Microbiological pathogens: live
poultry considerations, in Poultry meat processing CRC Press Boca Raton
3. Kotula, K.I., Wang, Z., 1994 - Characterization of broiler meat quality factors as
influenced by feed withdrawal time. Journal of Applied Poultry Research, 3, 103
4. Northcutt, J., 2001 - Preslaughter factor affecting poultry meat quality in Poultry
meat processing CRC Press Boca Raton, London, New - York, 5-19
5. indilar E., Greceanu Al., Carp-Crare M., Negrea A., Bondoc I, 1993 - Unele
aspecte privind reducerea gradului de poluare microbiana a crnii de pasre.
Lucrari stiintifice I.A.I., vol. 35-36, 101-106
6. Warriss, P.D., Wilkins, L.J., Knowles, T.J., 1999 - The influence of ante-mortem
handling on poultry meat quality in Poultry Science Symposium, vol. 25, CABI
Publishing, U.K.

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IMPLEMENTAREA PROCEDURILOR DE IGIENIZARE


N VEDEREA CERTIFICARII SISTEMULUI
DE MANAGEMENT AL SIGURANEI ALIMENTELOR
N UNITI DE PRELUCRARE A PETELUI
ADOPTING SANITATION PROCEDURES IN ORDER TO CERTIFY
FOOD SAFETY MANAGEMENT SYSTEM IN FISH PROCESSING UNITS

Carmen PETCU, C. SAVU, L. ILIE,


Elena MITRNESCU, Camelia PAPUC
U.S.A.M.V. Bucureti

The sanitation of production areas in fish processing units holds an


important role in assuring safe products for the consumers. In order to
prove that one has the knowledge of sanitation practices, there must be
developed a specific protocol or a handbook for good hygienic practices,
which must include the following: production and storing areas; equipment
and packaging materials; personnel as well as working and protection
equipment; facilities (looker rooms, sinks, toilets, working equipment);
means of transportation auto vehicles; infrastructure (buildings, water
supply network, and for evacuating residual, technological and disposable
water); yard [1, 3]. Also, proper personnel instruction must be done and
proved through providing a signed form; the proof of strict implementation
of this procedure consists of registered adequate statements.

MATERIAL I METOD

Procedura general ntocmit pentru meninerea sub control a strii de


igien, vizeaz toate locaiile i cuprinde modul de igienizare pentru: spaii de
producie i depozitare; utilaje i ambalaje; personal i echipament de lucru i
protecie; faciliti (vestiare, chiuvete, toalete, ustensile); mijloace auto de
transport; infrastructur (cldire, reelele de alimentare cu ap i de evacuare ape
reziduale tehnologice i menajere); curte [1, 3].
n vederea ntocmiri unei proceduri de igienizare sunt necesare o serie de
documente de referin i conexe.
Documente de referin pot fi reprezentate de: SR EN ISO 9000 : 2001 -
Sisteme de management ale calitii. Principii fundamentale, vocabular; SR EN
ISO 9001 : 2001 Sisteme de managementul calitii. Cerine; DS 3027 E : 2002
Cerine privind sistemul de securitate alimentar conform principiilor HACCP;
SR EN ISO 22000 : 2005 - Sisteme de management al siguranei alimentelor.
Cerine pentru orice organizaie din lanul alimentar [5, 6].
Documentele conexe vizeaz o serie de normative legale i de
reglementare care includ H.G. 1198/oct 2002 Privind aprobarea Normelor de
igien a produselor alimentare, OMS 976/1998 pentru aprobarea normelor de
igien privind producia, prelucrarea, depozitarea, transportul i desfacerea

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alimentelor [7]; Ghidul privind curenia, dezinfecia i sterilizare n unitile de


asisten medical Institutul de Sntate Public Bucureti; OMSF 933/2002;
H.G. 924/2005 privind aprobarea Regulilor generale pentru igiena produselor
alimentare; H.G. nr. 954/2005/CE privind aprobarea Regulilor specifice de igien
pentru alimente de origine animal care transpune n legislaia naional
Regulamentul 853/2004/CE, publicat n Monitorul Oficial nr. 805/05.09.2005 etc.
Prin documentarea i aprobarea unei proceduri privind igienizarea sunt
stabilite responsabiliti i limite de execuie a manoperelor necesare inerii sub
control a condiiilor de igien.
Toate procedurile din unitatea de producie vor fi codificate conform
Procedurii de control a documentelor din organizaie. Procedura privind
igienizarea va fi identificat astfel: PG (Procedur General) IG (Igienizare)
09 (numrul de ordine al procedurii), iar formulare vor prelua codul procedurii
care l-a introdus, urmat de litera F (Formular) i dou cifre ce vor reprezenta
numrul de ordine al formularului ntocmit. Ex.: PG-IG-09-F-01 este primul
formular introdus de procedura de igienizare cu numrul 09 din cadrul Sistemului
de management al siguranei alimentelor HACCP.
Managementul unitii stabilete, prin decizie sau prin trecerea
responsabilitilor n Fia de post, un responsabil care urmrete buna
funcionare a reelelor de evacuare a apelor reziduale tehnologice i menajere.
Acesta completeaz fia de urmrire a reelelor de evacuare ape reziduale i
dispune corecii sau aciuni corective n cazul apariiei defeciunilor, precum i
aciuni preventive pentru evitarea apariiei acestora.
La recoltarea probelor de ape uzate este necesar prezena unui
reprezentat al unitii care completeaz registrul de recoltri, iar buletinele de
analiz se arhiveaz i se pstreaz un timp stabilit conform procedurii care
reglementeaz pstrarea nregistrrilor.
Igiena infrastructurii se va realiza conform programului de igienizare, iar
persoana desemnat ca responsabil, va urmri realizarea acestuia i va stabili
materialele, modul i momentul igienizrii infrastructurii astfel nct s nu fie
afectat procesul de producie.
Igiena utilajelor, a instalaiilor tehnologice i a ambalajelor se realizeaz cu
respectarea Regulilor de buna practica pentru igienizare a acestora. Se va stabili
frecvena cu care se igienizeaz fiecare utilaj, responsabilul cu igienizarea,
substana folosit, concentraia substanei, modalitatea de utilizare (timp de
aciune, timp de contact cu suprafeele, temperatura apei folosit la diluii,
conduita de dup aplicare etc).
Medicul veterinar al unitii de producie organizeaz i verific fia de
urmrire zilnic a strii de igien a utilajelor i facilitilor, cod PG-IG-09-F-01,
completat de responsabilul desemnat, instruit n acest sens i dispune corecii i
aciuni corective n cazul n care utilajele nu sunt igienizate corespunztor; asist
i nscrie n registrul de recoltare a testelor de sanitaie prelevarea de ctre
organele de control abilitate a acestora i va urmri rezultatele [2, 3].
Este necesar dovada de inere sub control a strii de igien i sntate a
personalului prin ncheierea unui contract cu un medic de medicina muncii i prin
efectuarea analizelor medicale periodice, dovedite prin meninerea la zi a
carnetelor de sntate pentru fiecare angajat n parte. Mai mult dect att, un

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responsabil stabilit i instruit corespunztor, verific starea de igien i sntate a


personalului i completeaz fia zilnic de urmrire, cod. PG-IG-09-F-02. n cazul
n care se constat nereguli, lucrtorul respectiv nu va fi admis la lucru n spaiile
de producie sau depozitare.
inerea sub control a strii de igien a mijloacelor de transport auto se
realizeaz conform programului de igienizare a mijloacelor auto i cu respectarea
prevederilor OMS 976/1998-art.43 [4, 7]. oferul are obligaia de a prezenta
mijlocul auto la rampa de igienizare conform programului de igienizare, iar dup
efectuarea igienizrii, va prezenta executantului igienizrii carnetul cu evidena
igienizrilor, pentru al tampila. Se poate accepta orice modalitate de lucru care
s dovedeasc faptul c aciunile de igienizare sunt efectuate, iar corectitudinea
manoperei este verificat.
Este necesar stabilirea unei persoane care face selecia furnizorilor i
substanelor dezinfectante, recepia substanelor dezinfectante i apoi
completeaz registrul intrrilor de substane chimice de igienizare. Aceeai
persoan sau un alt reprezentat este instruit cu privire la modul de preparare a
soluiilor (conform instruciunilor de folosire) pe care le distribuie ori de cte ori
este nevoie, verificarea acestuia fiind efectuat de un specialist.
Ustensilele necesare aciunilor de igienizare din spaiile de producie sunt
identificate i depozitare ntr-un loc special amenajat. De asemenea, nu trebuie
omis o instruciune de lucru care sa stabileasc modalitatea de igienizare a
ustensilelor folosite la igienizare.

REZULTATE I DISCUII

Meninerea strii de igien a spaiilor de lucru sau depozitare, a facilitilor,


utilajelor, ustensilelor, precum i starea de igien a personalului angajat poate fi
meninut i dovedit prin documentarea unei proceduri generale de igienizare
(PG-IG-09) i meninerea unor nregistrri care s fac dovada efecturii
aciunilor ntreprinse.
n toate unitile de producie unde se dorete implementarea i certificarea
sistemului de management al siguranei alimentare sunt stabilite atribuii i
responsabiliti pentru toi angajaii, ncepnd cu managementul de vrf i pentru
toate activitile necesare.
Managementul de vrf aprob programul de igienizare curte i vecinti
ale tuturor locaiilor; aprob programul de igienizare infrastructur; aprob
programul de igienizare a utilajelor i ambalajelor; necesarul de substane chimice
de igienizare i ustensile de igienizare; aprob programul de igienizare faciliti;
aprob programul de igienizare spaii de depozitare i producie; aprob
programul de igienizare mijloace auto i aloc fondurile necesare bunei
desfurri a acestor activiti.
Echipa de igienizare sau un responsabil desemnat, ntocmete programul
de igienizare curte i vecinti ale tuturor locaiilor; distribuie n toate locaiile
programul aprobat de igienizare curte i vecinti; solicit necesar de ustensile i

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substane pentru realizarea programului de igienizare curte i vecinti; numete,


instruiete i verific realizarea programului de igienizare a curii i vecintilor
seciei.
Medicul veterinar ntocmete programul de igienizare a utilajelor i
ambalajelor; ntocmete programul de igienizare al mijloacelor auto; stabilete
substanele chimice de igienizare de achiziionat; vizeaz i supune aprobrii
necesarul de substane chimice de igienizare i ustensile de igienizare; ntocmete
programul de igienizare faciliti; ntocmete programul de igienizare spaii de
depozitare i producie; instruiete personalul i verific realizarea programului de
igienizare a utilajelor i ambalajelor; ine sub control carnetele de sntate a
personalului,verific starea de igien i sntate a personalului i completeaz fia
de urmrire etc.
eful de secie organizeaz activitatea de igienizare a echipamentelor de
lucru i protecie; verific zilnic starea de igien a facilitilor i completeaz fia
de urmrire zilnic a strii de igien a facilitilor; instruiete i verific efectuarea
programului de igienizare spaii de producie i depozitare;.
Responsabilul cu igienizarea gestioneaz i completeaz registrul
intrrilor de substane chimice de igienizare; completeaz registrul preparrilor
soluiilor de igienizare; completeaz fia de urmrire zilnic a strii de igien a
utilajelor i ambalajelor; solicit necesar de substane chimice de igienizare i
ustensile de igienizare etc.
Pentru a exista dovada ca ceea ce este scris n proceduri i instruciuni de
lucru se respecta n practic, se fac nregistrri ale tuturor activitilor efectuate,
fiind ntocmite fise specifice fiecrei activiti n parte.

CONCLUZII

1. Pentru implementarea sistemului de management al siguranei alimentare


n uniti de prelucrare a petelui se documenteaz o procedur general de
igienizare.
2. Managementul de vrf al organizaiei trebuie s susin i s aloce
resurse materiale, umane i de timp, necesare implementrii prevederilor
procedurii stabilite.
3. Toi angajaii din unitatea de prelucrare a petelui trebuie s fie instruii,
s cunoasc i s respecte prevederile procedurii de igienizare din societate.
4. Pentru fiecare unitate de industrie alimentar, procedura de igienizare
este specific, iar nregistrrile care se efectueaz sunt o dovada a respectrii celor
nscrise n procedura documentat.

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BIBLIOGRAFIE

1. Bratu Iuliana, Spulber Graziella, Ioarga A., 2002 Controlul calitii i HACCP n
industria alimentar, Editura Universitii Lucian Blaga, Sibiu;
2. Mitrea I.S., Carmen Petcu, G. Savu, 2003 Sigurana alimentelor prin aplicarea
sistemului HACCP, Ed. Bogdana;
3. Savu, C., Carmen Petcu, 2002 Inspecia i controlul produselor i alimentelor de
origine animal, Ed. Semne;
4. DS 3027E, 2002 Securitatea alimentului conform HACCP (Analiza riscului i
punctele critice de control) cerine care trebuie satisfcute de companiile
productoare i contractanii lor;
5. H.G. 924/2005 - privind aprobarea Regulilor generale pentru igiena produselor
alimentare;
6. H.G. nr. 954/2005/CE - privind aprobarea Regulilor specifice de igien pentru
alimente de origine animal care transpune n legislaia naional Regulamentul
853/2004/CE, publicat n Monitorul Oficial nr. 805/05.09.2005
7. SR EN ISO 9000, 2000 - Sisteme de management al calitii Noiuni fundamentale
i vocabular;
8. R EN ISO 22000, 2005 - Sisteme de management al siguranei alimentelor. Cerine
pentru orice organizaie din lanul alimentar

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EVOLUIA I SEMNIFICAIA NUMRULUI


I CONFIGURAIEI CELULELOR DIN LAPTE
LA OAIE I CAPR
THE EVOLUTION AND SIGNIFICANCE OF THE SOMATIC CELL NUMBER
AND CONFUGURANCE FROM GOAT AND SHEEP MILK

Diana SABU*, O. ROTARU


U.S.A.M.V. Cluj-Napoca

After somatic cells in milk have been found with almost a century
ago, they have been considered as being part of the small milk components.
The cytomorphological study of the milk sediment stained May-Grunwald-
Giemsa, and the somatic cell count using ISO-13366-1 method applied on
50 goat and 50 sheep raw milk samples, gathered thru the whole lactating
period, permitted us to establish with a great accuracy the hygienic quality
of it. The somatic cell count differs from the European Community
standards, in the case of 78% of goat milk samples and 72% for the sheep
milk samples. The cytologic configuration of the sediment smears made
possible to differentiate the following milk categories: healthy, normal milk,
milked and presented in hygienic conditions; highly microbiologically
polluted milk; milk from animals in the final period of lactation and
mammary pause; milk from animals with chronic or subacute mastitis,
animals which are carrying and eliminating germs.

INTRODUCERE

Atenia asupra configuraiei populaiei celulare din lapte i mai ales variaia
lor ca numr, a suscitat un mare interes atunci cnd s-a demonstrat o relaie
direct ntre numrul lor i evoluia la nivelul mamelei, a unor procese
inflamatorii (mamite) predominent exudative, care pot compromite grav calitatea
igienic a laptelui, ca produs alimentar. n prezent, sunt stabilite limite ale
numrului de celule pe mililitru lapte, pe baza crora se stabilete calitatea
laptelui i care se iau n considerare la stabilirea preului de achiziie al acestuia,
de la productor (3, 4).

MATERIAL I METOD

Fr a minimaliza importana determinrii numrului total de celule, inclusiv


prin metode electronice, studiul citologic al laptelui, att pe frotiuri realizate din
lapte ca atare, ct mai ales din lactosediment, poate releva mult mai multe detalii

*
E-mail: didisab@clujnapoca.ro

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legate de msurile care se impun n legtur cu valorificarea laptelui n consumul


uman.
Pentru a testa valoarea examenului citologic n supravegherea strii de
sntate al acestuia, s-a procedat la recoltarea de probe de lapte proaspt (crud)
de la o populaie de caprine i ovine din judeul Cluj. Au fost prelevate 50 probe
lapte capr i 50 probe de lapte oaie, pe ntreaga perioad de lactaie (1 aprilie
20 august 2005), la care s-a determinat numrul celulelor somatice prin metoda
ISO-13366-1, rezultatele fiind corelate cu examenul citologic pe frotiuri fixate i
colorate prin metoda May-Grnwald Giemsa.

REZULTATE I DISCUII

Trebuie s precizm de la bun nceput, c toate probele examinate au avut


nsuiri senzoriale i fizico-chimice, caracteristice de lapte normal, sntos, care
se admite n consum fr restricie, sub rezerva tratamentului termic (pasteurizare,
fierbere) nainte de a fi consumat.
Metoda de referin la noi n ar pentru numrarea celulelor somatice din
lapte este metoda microscopic ISO-13366-1, standard identic cu norma
internaional ISO-13366-11996 Lait-Denombrement des cellules somatiques,
Partie 1: Methode au microscope (Methode de reference). Prezentul standard
corespunde de asemenea i Directivei CE 91/180/CE din 14.02.1991.
Numrarea celulelor somatice este o metod de baz n programele de
control a laptelui. Comunitatea European a stabilit limita legal pentru laptele de
vac, dar fr a face referiri i la laptele de oaie i capr, n Directiva EEC/92/46
(1992). Aceast situaie a fost temporar, deoarece UE a sugerat stabilirea pentru
viitor a unei limite pentru celulele somatice i la rumegtoarele mici.
n SUA, limita legal pentru numrul celulelor somatice la laptele de capr
a fost stabilit la 1.000.000 celule/ml i pentru oaie la 750.000 celule/ml, iar
cresctorii frecvent nu pot respecta aceast limita (2). La Simpozionul
internaional Somatic Cells and Milk of Small Ruminants (Bella, Italia, 1993), s-a
sugerat autoritilor Comunitii Europene ca pragul maxim al celulelor somatice
pentru laptele de colectur de capr i oaie s fie de 1.500.000 celule/ml (1).
Numrul de celule somatice (SCC Somatic cells count) obinut, depete
limita maxim admis la 78% din probele de lapte de capr i la 72% la cele de
oaie. Cu toate acestea, examenul citomorfologic a pus n eviden o proporie mai
mic a laptelui de oaie cu caracter mamitic, fapt corelat i cu rezultatele
numrtorii celulare unde valorile celulelor somatice sunt doar uor crescute peste
standardul de 750.000 celule/ml.
Stadiul lactaiei influeneaz numrul de celule somatice si indic faptul c
SCC crete odat cu vrsta i stadiul lactaiei, dar modificrile sunt reduse la
animalele sntoase.
Argumentele citomorfologice care au permis identificarea urmtoarelor
categorii de lapte:

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Lapte cu aspect normal, unde populaia celular este puin reprezentat,


celulele monocite macrofage nu sunt activate, unele fiind transformate n
lipofage; PMN sunt rare, vrstnice, cu nucleul intens segmentat, iar celulele
epiteliale se gsesc n diferite stadii de necrobioz (Fig. 1).

Fig. 2 Citosediment lapte de capr


Fig. 1 Citosediment lapte de oaie sntos
normal, sntos: rare lactocite i resturi de
monocit macrofag (coloraie MGG, x1000)
celule epiteliale (coloraie MGG, x1000)
Totui, la unele probe, s-au obinut valori ale SCC peste limitele admise,
dei nu au fost prezente semnele citomorfologice caracteristice unui proces
inflamator, examenul morfologic evideniind un numr mare de celule sferice, cu
nucleul periferic, semilunar - lactocitele caracteristice acestor specii.
Preponderena acestor celule n frotiurile de lapte, cu modificarea semnificativ a
numrului de celule somatice, dar fr reacia celular caracteristic procesului
inflamator, rmne o curiozitate, dar i un aspect care trebuie elucidat. Prezena n
laptele de capr, dar i n cel de oaie, a particulelor citoplasmatice, anucleate sau
nucleate, de diferite dimensiuni, mascheaz i fac dificil interpretarea
rspunsului leucocitelor la infecie, astfel nct diferenierea dintre celule ar putea
fi de interes n diagnosticul mastitei neclinice (fig. 2). Metodele de identificare a
leucocitelor sunt bazate n mod obinuit pe identificarea nucleelor.
Lapte poluat microbiologic, unde componenta celular nu reacioneaz
pentru fagocitarea germenilor, demonstrndu-se c este vorba de o contaminare a
laptelui n timpul mulsului i/sau recoltrii, dei la numrtoarea celular nu s-au
determinat abateri de la limitele maxime admise (Fig. 3). Se permite astfel
evidenierea i aprecierea riscului microbiologic prin identificarea pe frotiu a
microflorei prezente n lapte.
Lapte de retenie, unde populaia celular se concentreaz ntr-un volum
mic de lapte, cum este cel din finalul lactaiei, caracteristicile morfologice ale
celulelor nu difer n mare msur de cele prezente n laptele normalO
particularitate la laptele de la sfritul lactaiei, la ambele specii, este prezena
unui numr mare de monocite macrofage, dar care au activitate fagocitar fa de
structurile proprii, transformndu-se n lipofage (Fig. 4).

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Fig. 4 Citosediment lapte de capr, lapte


Fig. 3 - Citosediment lapte de oaie
de retenie Monocit macrofag hiperactivat
poluare microbiologic polimorf (coloraie
pe care ader o celul epitelial (coloraie
MGG, x1000)
MGG, x1000).

Laptele mastitic, a format dup centrifugare un sediment mai abundent,


vscos, cu aspect filant, populaia celular fiind crescut numeric, predominnd
celulele polimorfonucleare i prezena de microparticule de exudat purulent n
mamitele cu evoluie acut i subacut si celulele monocite-macrofage activate i
hiperactivate n cele cu evoluie cronic. n frotiurile din acest lapte, celulele au
tendina de a se grupa n conglomerate, cte 2-3, stabilindu-se legturi celulare
prin adevrate puni citoplasmatice (Fig. 5). n citoplasma multor
polimorfonucleare i macrofage, sunt prezeni microbi fagocitai, uurnd astfel
stabilirea i a unui diagnostic etiologic al procesului inflamator (Fig. 6).

Fig. 6 Citosediment lapte de capr,


Fig. 5 Citosediment lapte de capr, mamit supurat strepto-stafilococic
mamit supurat cu microcoaguli de puroi numeroase PMN i macrofage hiperactivate
(coloraie MGG, x1000) i microconglomerat purulent (coloraie
MGG, x1000).

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CONCLUZII

Alturat metodei de numrare a celulelor, citodiagnosticul pe lactosediment


poate constitui o metod sigur de diagnostic, pentru supravegherea calitii
igienice (sntii) laptelui. Citodiagnosticul are fa de numrarea celulelor i
cteva avantaje, printre care mai importante sunt: posibilitatea de a diferenia un
lapte mastitic fa de un lapte colostral sau la final de lactaie, identificarea
laptelui contaminat microbiologic i nu n ultimul rnd, poate s precizeze dac
laptele este obinu sau nu n condiii igienice. Mai mult, chiar examenul citologic
poate de multe ori stabili etiologia procesului inflamator naintea rezultatului
examenului microbiologic prin identificarea morfologic a microflorei prezente n
microparticolele de exudat fibrinopurulent i mai ales a microbilor fagocitai n
citoplasma polimorfonuclearelor neutrofile i monocitelor macrofage.

BIBLIOGRAFIE

1. Barbosa M., Miranda R., 1984 Addendum to special publication No. 49. Changes
to Contemporary Dairy Analytical Techniques, Royal Society of Chemistry,
London, UK
2. Droke E.A., Paape M.J., Di Carlo A.L., 1993 Prevalence of high somatic cell
countc in bulk tank goat milk, Journal Dairy Sci. 76: 1035 1039
3. Park Y.W., 1991 Interrlationships between somatic cell counts electrical
conductivity, bacteria counts, percent fat and protein in goat milk, Small
Ruminant Research 5; 367-375. Elsevier Science Publishers B.V.Amsterdam.
4. Rotaru O., L. Ognean, 1998 Morfologia i fiziologia populaiei celulare din lapte.
Ed.Casa Crii de tiine, Cluj-Napoca.
5. Directiva CE 91/180/CE din 14.02.1991
6. Directiva EEC / 92 / 46 1992

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CALITATEA LAPTELUI - PARTE A CONCEPTULUI


,,SIGURANA ALIMENTELOR N PERSPECTIVA
INTEGRRII EUROPENE
THE QUALITY OF RAW MILK - PART OF ,,FOOD SAFETY CONCEPT,
PROSPECT FOR E.U. INTEGRATION

Claudia Gabriela DUMINIC


D.S.V.S.A. L.S.V.S.J. Iai
V. FLORITEAN, E. INDILAR
U.S.A.M.V. Iai

Milk is sterile at secretion in the udder but is contaminated by


bacteria even before it leaves the udder. Except the case of mastitis, the
bacteria at this point are almost always harmless and few in number.
Further contamination of the milk with microorganisms can take place
during milking, handling, storage, and other pre-processing activities.
Information on the microbial content of milk can be used to judge its
hygienic quality and the conditions of production. If permitted to multiply,
bacteria in milk can cause spoilage of the product.
Milk is potentially susceptible to contamination with pathogenic
microorganisms. Precautions must be taken to minimize this possibility and
to destroy pathogens that may gain entrance. Milk produced from
conventional milk works, implying the production of a more hygienic
product.

Key words: raw milk quality, milk hygiene, somatic cell count, E.U. integration,
food safety

Riscul alimentar pentru om, conform evalurii experilor, este determinat,


n proporie de 90% de alimentele de origine animal. (2, 3, 5, 7)
Experiena comunitar a demonstrat c economia de pia postindustrial,
caracterizat prin ncurajarea iniiativei individuale, dezvoltarea unui sistem
concurenial i stabilirea prghiilor economico-financiare, conduce la realizarea
de producii animaliere peste nivelul necesar (cantitativ) al populaiilor umane din
rile respective i creeaz disponibiliti mari pentru export. (1, 8)
Nu s-a rezolvat ns, problema siguranei alimentelor pentru populaie, fapt
ce a determinat crearea i dezvoltarea unei autoriti reale - Autoritatea
Europeana pentru Sigurana Alimentelor (E.F.S.A.) cu competene n: evaluarea
riscului alimentar, gestiunea (managementul) riscului alimentar, comunicarea
riscului alimentar organizarea sistemul rapid de alert (S.R.A.), care presupune:
prevenirea ptrunderii pe pia sau returnarea din reeaua comercial a alimentelor
de origine animal cu risc major pentru sntatea consumatorilor, prevenirea

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ptrunderii n consumul animalelor a unor furaje care pot afecta indirect sntatea
consumatorilor, informarea rapid, la cele 3 nivele (central, judeean i local)
asupra problemelor sau riscurilor cu privire la producerea alimentelor de origine
animal i la furajele care nu ndeplinesc cerinele impuse de sigurana alimentar.
Prin intermediul S.R.A. pot fi transmise urmtoarele categorii de risc:
modificri organoleptice, limite microbiologice depite, infestaii parazitare,
contaminri cu reziduuri i elemente legate de falsificri, ambalare sau etichetare.
Astfel, s-a concis c gestiunea riscului alimentar are cele mai mari anse s
fie realizat sub coordonarea serviciilor veterinare, conform principiului ,,de la
furc la furculisau de la ferm la masa consumatorului, pentru a se asigura
trasabilitatea produselor i pentru a se garanta n cele din urm eliminarea riscului
alimentar.
Guvernul Romniei a adoptat strategia Comunitii Europene n domeniul
siguranei alimentare i a nfiinat Agenia Naional Sanitar Veterinar i pentru
Sigurana Alimentelor - Instituie public cu personalitate juridic Organism de
specialitate al administraiei publice centrale - Autoritate a statului n domeniul
veterinar i al siguranei alimentelor.
Date citate de A.N.S.V.S.A. relev faptul c numai 6% din totalul de
uniti sunt conforme cu normele U.E., n vreme ce 17% dei nu ndeplinesc
condiiile impuse au ansa, ca prin investiii, si desfoare activitatea i dup 1
octombrie 2006.
Unitile care proceseaz lapte au beneficiat de derogri de pn la 2 ani,
cei care nu au posibilitatea de investiii n profilul de activitate desfurat, -i pot
schimba profilul de activitate n uniti cu vnzare direct sau pot devenii fabrici
de produse tradiionale.
n domeniul igienei i sntii publice, unul dintre obiective l reprezint
alinierea standardelor de igiena ale unitilor din industria alimentara la
cerinele comunitare.
n lucrarea de fa ne-am propus s evideniem unele corelaii ntre
numrul de celule somatice i valorile unor parametri microbiologici pentru
probele de lapte analizate de la centrele de colectare sau de la unitile de
procesare lapte i evoluia acestora n vederea restructurrii, modernizrii ca scop
al ncadrrii n uniti din categoria A, care ndeplinesc condiiile pentru
desfurarea activitii conform normelor impuse de legislaia U.E., n uniti din
categoria B care s-au angajat ca pn n decembrie 2006 vor finaliza programele
de modernizare restructurare, n uniti de categoria C, care au primit derogare
pn n 2009 pentru a-i realiza programul de modernizare restructurare i
uniti de categoria D, care vor fi supuse lunar evalurilor din partea instituiilor
cu atribuii n acest sens.

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MATERIAL I METOD

n perioada ianuarie decembrie 2005 au fost investigate, n scopul


evalurii calitii, un numr de 485 probe lapte materie prim, recoltate n cadrul
Programului strategic (304 probe) i n cadrul Programului de autocontrol (181
probe).
Probele investigate provin cu preponderen de la 15 centre de colectare
lapte materie prim din judeul Iai, 8 aparinnd societii A, 3 aparinnd
societii B, 3 aparinnd societii C i unul aparinnd societii D. La acestea s-
au adugat un numr mai mic de probe care au fost recoltate direct de la unitile
de procesare, din cisternele de transport, i probe recoltate de ctre medicii
concesionari de la animale din gospodriile populaiei.
Probele recoltate din centrele de colectare i cele de la unitile de
procesare lapte au fost, cu precdere, probe de lapte amestec n timp ce probele
recoltate din gospodriile populaiei au fost reprezentate de probe individuale.
Probele investigate au corespuns din punct de vedere al aspectelor
senzoriale i a strii de prospeime.
Pentru determinrile efectuate s-au respectat toate exigenele referitoare la
condiiile de recoltare, reprezentativitate, manipularea i pstrarea probelor.
Determinarea numrului total de celule somatice s-a efectuat n
conformitate cu SR ISO 13366/1/1997 cu ajutorul analizorului CombyScope,
alctuit din: unitate de procesare prob Sample Preparation Unit, Soma Scope
MK 2 unitate de evideniere numrare i imprimanta matricial.
Pentru determinrile microbiologice s-au utilizat metodele impuse de
Asociaia Internaional de Standardizare, dup cum urmeaz:
SR ISO 6610/1997 pentru determinarea N.T.G. stabilirea numrului
de microorganisme, metoda prin numrarea coloniilor obinute la 30C,
SR ISO 5541-1/94 pentru determinarea bacteriilor coliforme din lapte
i produse lactate,
SR ISO 5541-2/94 pentru stabilirea numrului de bacterii coliforme -
Metoda prin numrarea coloniilor la 30oC i tehnica numrului cel mai probabil
(MPN) la 30oC,
SR ISO 7251/1996 pentru stabilirea numrului de Escherichia coli
prezumtiv - Tehnica numrului cel mai probabil,
SR EN ISO 6888/1/2/2002 pentru determinarea stafilococilor
coagulazo-pozitivi din lapte i produse lactate,
SR ISO 6579/2003 pentru determinarea prin metod orizontal a
bacteriilor din genul Salmonella, din lapte i produse lactate,
SR ISO 7923/2003 pentru determinarea bacteriilor din genul Bacillus
cereus, din lapte i produse lactate - Stabilirea numrului de B. cereus, metoda
numrrii coloniilor la 30oC,
SR ISO 11290/1/2/2000 pentru determinarea bacteriilor din genul
Listeria, din lapte i produse lactate, metoda orizontal pentru detectarea i
numrare.
Identificarea bacteriilor sporulate anaerobe din lapte sa efectuat prin
proba Weinzirl, conform metodei descrise n STAS 6349-5/80.

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Pentru efectuarea analizelor s-au utilizat medii de cultur deshidratate,


medii cu adaosuri de reactivi sau ingrediente specifice conforme cu metodologia
impus de standardele menionate , reactivi i antiseruri specifice provenite din
import, dar i produse biologice autohtone.
Interpretarea rezultatelor obinute s-a fcut n conformitate cu Ordinul
Ministerului Sntii 975/1998 si Ordinul M.A.P.M 1106/2003 (etapa a II a
perioada 1 ianuarie 2005 31 decembrie 2006)

REZULTATE I DISCUII

Au fost efectuate, n total, un numr de 2144 analize constnd n examene


microbiologice i determinarea numrului total de celule somatice (N.C.S.) din
lapte.
Determinarea numrului total de germeni (N.T.G.) s-a efectuat la un
numr de 416 probe de lapte materie prim (din care 304 n Programul strategic i
112 probe n cadrul Programului de autocontrol)
Rezultatele examenelor de laborator au relevat ca necorespunztoare un
numr de 107 probe din probele examinate n cadrul Programului strategic
(35,2%) i 43 probe examinate n cadrul Programului de autocontrol (62,3%).
Probele ncadrate ca fiind necorespunztoare au valori ale parametrului
N.T.G./ ml ncadrate ntre 1.000.000 i 2.480.000 germeni/ml.
Determinarea bacteriilor coliforme /ml s-a efectuat la un numr de 375
probe lapte materie prim (304 probe recoltate n cadrul Programului strategic
i 71 probe n cadrul Programului de autocontrol) din care un numr de 141
37,6% (112 probe 29,9% recoltate n cadrul Programului strategic i 29 probe
7,7% n cadrul Programului de autocontrol) sunt ncadrate ca fiind
necorespunztoare, valorile obinute fiind superioare celor admise de
O.M.S./975/1998.
Analiza probelor pentru decelarea Escherichia coli/ml relev c dintr-un
numr de 366 probe lapte materie prim, (294 probe recoltate n cadrul
Programului strategic i 72 probe n cadrul Programului de autocontrol), un
numr de 103 28,14%, (77 probe 21,03% recoltate n cadrul Programului
strategic i 26 probe 7,1% n cadrul Programului de autocontrol) au fost
necorespunztoare, valorile obinute fiind superioare celor admise de
O.M.S./975/1998
Laptele recoltat de la animale sntoase, exploatate n condiii de igien,
muls n condiii de igien desvrite, conine un numr de microorganisme mic,
ntre 300 i 500/ml n timp ce laptele muls de la animale cu diferite infecii
mamare, exploatate n mod necorespunztor, la care manoperele de muls sunt
efectuate n condiii precare de igien numrul de microorganisme crete pn la
ordinul sutelor de mii i chiar al milioanelor/ml.
n ceea ce privete parametrul microbiologic Stafilococi coagulazo-
pozitivi/ml, examinarea probelor a relevat c dintr-un numr de 344 probe lapte

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materie prim (294 probe recoltate n cadrul Programului strategic i 40 probe


n cadrul Programului de autocontrol), un numr de 8 probe 2,32% (6 probe
1,74% recoltate n cadrul Programului strategic i 2 probe 7,1% n cadrul
Programului de autocontrol) au fost ncadrate ca fiind necorespunztoare,
valorile obinute fiind superioare celor admise de O.M.S./975/1998
Analizele pentru decelarea parametrului Bacillus cereus/ml relev c
pentru un numr de 122 probe lapte materie prim, recoltate n cadrul
Programului strategic rezultatele au fost corespunztoare limitelor admise de
O.M.S./975/1998.
Analizele pentru decelarea parametrului Salmonella/25 ml relev c pentru
un numr de 304 probe lapte materie prim, recoltate n cadrul Programului
strategic rezultatele sunt ncadrate ca fiind corespunztoare, limitelor admise de
O.M.S./975/1998.
Analizele pentru decelarea parametrului Listeria/25 ml c relev c pentru
un numr de 128 probe lapte materie prim, recoltate n cadrul Programului
strategic rezultatele sunt ncadrate ca fiind corespunztoare, limitelor admise de
O.M.S./975/1998.
Bacteriile anaerobe sporogene, productoare de gaz, au fost identificate la
un nivel ridicat n 13 (8,55%) probe din cele 152 probe materie prim la care s-a
efectuat acest parametru i au fost ncadrate ca fiind necorespunztoare, conform
condiiilor impuse de STAS 6349-5/80. (Tabel 1)
Tabelul 1
Investigaii privind calitatea microbiologic a laptelui materie prim

Anul Parametrii microbiologici analizai


2005 NTG B.colif. E.coli. Staph.cp. B.cereus Salm.* Listeria* Proba
Numr /ml /ml /ml /ml /ml /25ml /25ml Weinzirl
probe C N C N C N C N C N C N C N C N
P.S.
197 107 192 112 217 77 298 6 122 0 304 0 128 0 72 8
304
P.A.
69 43 42 29 46 26 38 2 0 0 0 0 0 0 67 5
181
*determinarea face referire la parametrii care nu au fost decelai, din motive tehnice,
pentru toate probele (referire la programul de autocontrol)

Numrul total de celule somatice N.C.S./ml a fost determinat la 485 de


probe, din acestea la 21,03% (85 probe recoltate n Programul strategic i la 17
probe recoltate n cadrul Programului de autocontrol) valorile obinute s-au
ncadrat n intervalul 600.000. 3.400.000 NCS/ml, fiind necorespunztoare, la
53,20% din probe s-a obinut valori cuprinse ntre 400.000 i 600.000 N.C.S./ml
iar la 25,77% din probe celulele somatice au fost sub 400.000/ml limita actual
fiind de maxim 600.000/ml. (Tabel 2)

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Tabelul 2
Investigaii privind valoarea numeric a celulelor somatice
din laptele materie prim

NCS**/ml
Anul Numr
2005 probe Corespunztoare
Necorespunztoare
a b
56 163
P.S. 304 85
219
69 95
17
P.A. 181 164
383 102

** valori care se ncadreaz astfel: a -<400.000, b - 400.000 - 600.000, c - >600.000 i se


raporteaz conform etapizrii specificate n Ordinul 1106/2003 al MAPM
** ncepnd cu anul 2004 valorile se consider necorespunztoare pentru probele care au
NCS/ml: >600.000 cu raportare la condiiile impuse de Ordinul 1106/2003 al MAPM
PS program strategic, PA program autocontrol

Numrul de celule somatice /ml se situeaz n jurul valorii de 80.000 /ml.


pentru o vac sntoas, intervalul dintre 80.000/ml. i 300.000/ml. ncadreaz
laptele ntr-un lapte dubios iar limitele cuprinse ntre 300.000/ ml. i 1.000.000/
ml. duce la aprecierea laptelui respectiv ca fiind suspect. (2, 6, 9)
Elementele celulare sunt reprezentate de celulele epiteliului mamare sau
fragmente ale acestora n procent de 60%- 80%, celule de origine sangvin n
procent de 20% -30%, limfatic i microorganisme. (2, 6, 9)
Celulele epiteliale de origine mamar se regsesc n laptele animalelor
sntoase, recoltat n condiii corespunztoare de igien n cantitate neglijabil,
numrul acestora crescnd considerabil n cazul evoluiilor proceselor inflamatorii
de la nivelul glandei mamare. (2, 6, 9)
Numrul de celule din laptele vacilor cu uger sntos variaz n intervalul
de valori maxime - minime n funcie de ras, vrst, stare fiziologic a glandei
mamare i chiar de la acelai sfert de la o recoltare la alta. La vacile aflate la
prima lactaie valoarea numeric a celulelor somatice este mai mic, aceasta
crescnd progresiv cu numrul de lactaii.
Se consider c numrul de celule somatice crete ncepnd cu a III a
lactaie i datorit stresului de muls, iar colostrul i laptele din primele sptmni
de dup lactaie conine peste 10.000/ ml., la nceputul mulsului peste 100.000/
ml. i crete n primele 2-4 ore de la muls.
Valoarea N.C.S./ml lapte de amestec probele recoltate de la centrele de
colectare i unitile de procesare lapte, a prezentat variaii considerabile,
rezultatele fiind ncadrate ntre 186.000/ml i 824.000/ml.
Valoarea N.C.S./ml pentru laptele individual probe recoltate de la
animalele din gospodriile populaiei este considerabil mai mic, aceasta
ncadrndu-se ntre limitele, minim 3000/ml i 126.000/ml.
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Se constat de asemenea c valoarea numeric a parametrului N.C.S./ml


este mai crescut n perioada de var, fapt care se asociaz cu evoluia mamitelor
subclinice, cnd sau nregistrat valori ncadrate n intervalul 400.000 - 600.000
N.C.S./ml. Tot n sezonul cald sau determinat pentru un numr mic de probe,
valori ale N.C.S./ml cuprinse n intervalul 1.343.000 2.797.000, probe care
proveneau de al animale cu mastit.

Fig. 1 - Harta Unitilor de procesare i a Centrelor de colectare lapte - judeul Iai.


Unitii de procesare lapte
- Categoria B: Promilch, Strunga,
- Categoria D: SC Nicodan SA, SC Romlacta, SC, Trans -Gigel
Centre de colectare lapte:
- SC ISPA ECO SRL Romneti, Mirosloveti, Vereni, Erbiceni., Moca,
Cristeti, Lespezi, Cozmeti
- SC ILVAS Vaslui Probota, Crniceni, Trifeti,
- SC Lactis SA Crniceni, Cotnari, Ceplenia,1- Camitex Cristeti

CONCLUZII

Centraliznd rezultatele necorespunztoare putem concluziona c la nivel


de uniti de procesare lapte, probele necorespunztoare sunt n procent de
30,92% pentru parametrul N.T.G./ml, 20,97% pentru parametrul
Bacterii.coliforme./ml, 21,93% pentru parametrul Escherichia coli/ml, 1,64%
pentru parametrul Stafilococi coagulazo-pozitivi/ml, 21,03% pentru parametrul
N.C.S./ml, 2,68% pentru parametrul Bacterii anaerobe gazogene/ml, dintr-un
total de 485 probe analizate, ceea ce denot existena unor deficiene, n modul
su de recoltare i de manipulare a laptelui materie prim, n modul n care se
monitorizeaz starea de sntate a animalelor productoare de lapte, n modul n

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

care sunt respectate condiiile de igien i nu n cele din urm n modul n care
sunt aplicate msurile impuse de legislaia sanitar veterinar aflat n vigoare.
Evaluarea neconformitilor care s-au constatat la nivel de recoltare a
laptelui materie prim, corelate cu aprecierea amplasrii i compartimentrii, a
fluxului existent n centrele de colectare i n unitile de procesare a laptelui, i
nu n cele din urm a performanelor utilajelor, n perioada analizat, a condus la
ncadrarea n judeul Iai doar a dou uniti de procesare a laptelui n categoria B,
a unui numr de trei uniti n categoria D iar pentru un numr de patru uniti de
procesare lapte s-a procedat la emiterea ordonanelor de suspendarea definitiv a
activitii.

BIBLIOGRAFIE

1. Apostu S., 2004 - Managementul calitii alimentelor, Ed. Risoprint, Cluj Napoca
2. Brzoi D., Apostu S., 2002 - Microbiologia produselor alimentare, Ed. Risoprint, Cluj
Napoca
3. Enache T., Paul I., Stnescu V., Popescu O., Iordache I., 2001 - Tratat complet de
medicin legal veterinar, Ed. All, Bucureti.
4. Enache T., Paul I.,, Stnescu V., Popescu O., Iordache I., 1995 - Medicin legal
veterinar. Ed.All, Bucureti.
5. Headrick M.L., Korangy S., Bean N.H., Angulo F.J., Altekruse S.F., Potter M.E.,
Klontz K.C., 1998 - The epidemiology of raw milk-associated foodborne disease
outbreaks reported in the United States, 1973 through 1992, Am. J. Public
Health; 88:1219-21.
6. Petkow P., Gergovkas J., and Radulova P., 2001 - The influence of some factors
on somatic cells count in cows milk. Zhivotnov deni-Nauki, 38 (2) 86-89.
7. Potter M.E., Kaufmann A.F, Blake P.A, Feldman R.A., 1984 - Unpasteurized milk:
the hazards of a health fetish. JAMA 252:2048-52.
8. Savu C., Georgescu N., 2004 - Sigurana alimentelor, riscuri i beneficii, Ed.
SemnE,. Bucureti
9. indilar E., 2000 - Controlul igienic al produselor i subproduselor de origine
animal, Vol. II, Ed. I.N.S.C.R. Iai,
10. **** Ordinul 975/1998 al M.S., Publicat n Monitorul Oficial, Partea I nr. 268 din
11/06/1999
11. **** Ordinul 1106/2003 al M.A.P.A.M., Publicat n Monitorul Oficial, Partea I nr. 61 din
23.01.2004
12. **** Standarde de stat, norme tehnice de calitate i de metode de analiza pentru
laptele integral materie prim: Stas 6349/ 5-8- 1980, SR ISO 6610/1997, SR
ISO 7923/2003, Norm sanitar veterinar preluat dup Directiva CEE
91/180,77/99, SR ISO 13366-1/ 98;
13. **** Standarde de stat, norme tehnice de calitate i metode de lucru pentru produse
lactate din lapte netratat termic: SR EN 12824/2001, SR ISO 11290/1/2000, SR
ISO 5541-1-2 /94, SR ISO 7251/96, SR ISO 6888/02, SR ISO 6579/2003

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Universitatea de tiine Agricole i Medicin Veterinar Iai

CONTROLUL STARII DE IGIEN I A CALITII APEI


N UNELE UNITI IEENE DE PRELUCRARE A
LAPTELUI, EFCTUAT PRIN TEHNICI CLASICE I RAPIDE
THE QUALITY HYGIENE CONTROL OF WORK, EQUIPAMENT OR HAND
SURFACES, PLAN ENVIROMENT AND WATER, IN SOME MILKWORKS FROM
IAI CITY BY USUAL AND FAST MICROBIOLOGICAL METHODS

Claudia Gabriela DUMINIC


D.S.V.S.A. L.S.V.S.J. Iai
E. INDILAR
U.S.A.M.V. Iai

This system, called Hazard Analysis and Critical Control Points -


HACCP, is a focus on critical milk safety areas as part of total quality
programs. It involves a critical examination of the entire milk
manufacturing process to determine every step where there is a possibility
of physical, chemical, or microbiological contamination of the food which
would render it unsafe or unacceptable for human consumption.
Colilert/Enterolert is an easy, rapid, and accurate method for the
detection of coliforms and E.coli/ enterococci in water.Colilert- 18/
Enterolert-E was developed for the European market and correlates with
the EU Drinking Water Directive standard method for coliforms and
enterococci, that gives you results in just 18 hours.
A new Positive Control Kit for validation of a number of their
hygiene and water monitoring devices, with systemSURE II ATP Hygiene
Monitoring System and Snapshot Universal ATP swab for facilities which
were previously using other luminometers. The Positive Control Kit allows
users to check accuracy of devices. A crucial part of Food Safety Progra. A
simple and fast way to validate the hygiene of a surface and demonstrate
due diligence in a HACCP program.
Slides for food and Water Screening, for the fast detection of
coliforms, faecal coliforms, faecal Streptococi and Pseudomonas in water.

Key words: quality hygiene control, work or equipament surfaces and waterin,
milkworks, etc.

Implementarea i aplicarea principiilor Sistemului HACCP i


contientizarea cresctorilor particulari individuali sau a celor organizai n ferme,
asupra importanei asigurrii asistenei sanitar veterinare pentru animalele
productoare de lapte, asupra recoltrii, manipulrii i procesrii laptelui n
condiii igienico-sanitare corespunztoare sunt condiii absolut eseniale pentru
obinerea unui produs finit de calitate, care s ndeplineasc cerinele
microbiologice impuse de U.E.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Starea de igien a materiei prime, a apei utilizate n procesul tehnologic,


starea de igien a utilajelor, ustensilelor i a personalului implicat se rsfrng
asupra calitii produsului finit.
n lucrarea de fa s-a urmrit evidenierea unor aspecte legate de calitatea
microbiologic a apei, a strii de igien pe fluxul tehnologic la nivelul a trei
uniti de procesare lapte.

MATERIAL I METOD

ntre ianuarie martie 2006 au fost investigate cu scopul determinrii


ncrcturii microbiologice a apei un numr de 63 probe ap.
Un numr de 9 probe recoltate n cadrul Programului strategic, la care sau
efectuat 45 analize i un numr de 28 probe pentru care 78 analize au fost
efectuate conform metodei de lucru impuse de Stas 3001.
Un numr de 16 probe, recoltate n conformitate cu Programul de
autocontrol, au fost investigate conform modului de lucru impus de testele rapide,
7 pe teste Colilert respectiv 9 pe teste Enterolert.
Un alt numr de 10 probe recoltate n conformitate cu Programul de
autocontrol, au fost investigate pe Slide food and water screening, cu un numr
de 30 determinri microbiologice.
Testele rapide Colilert respectiv Enterolert reprezint o metod
colorimetric/fluorimetric care permite detecia unor enzime int specifice
microorganismelor.
Substratul nutritiv indicator este metabolizat de ctre enzima int,
specific microorganismului i determin apariia unei culori caracteristice.
Metabolizarea microoganismelor asociate este inhibat.
Tehnica este rapida, rezultatele se obin n 24 de ore i se bazeaz pe
utilizarea din substratul nutritiv a indicatorului ONPG n cazul coliformilor, a
indicatorului MUG n cazul E.coli, i a indicatorului 4-methyl umbelliferyl -
D-glucoside pentru enterococi.
Se evideniaz prezena sau absena coliformilor prin apariia culorii
galbene, a E.coli. prin apariia culorii galbene fluorescente i a enterococilor
fecali prin apariia culorii albastre fluorescente.
Se poate executa i determinarea numrului cel mai probabil pentru
coliformi sau enterococi prin utilizarea godeurilor Quanti-Tray, a sigilatorului
Quanti-Tray sealer i a unei lmpi UV, cu interpretarea realizat prin utilizarea
tabelului MPN .

A B
Fig. 1. - Aprecierea calitativ a apei

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A B C

D E

Fig. 2 - Aprecierea cantitativ a ncrcturii apei n coliformi sau enterococi prin MPN

Pentru aprecierea calitii apei raportarea s-a efectuat la Legea 458/2003


completat cu Legea 311/2004 i un numr de 10 probe cu 30 de analize
raportate conform instruciunilor de lucru cu Slide food and water screening.
Slide food and water, screening sunt utilizate sub formule diverse de
detectie microorganisme cu: cod 71L52521 pentru NTG, drojdii i mucegaiuri,
Enterobacteriaceae, cod 71L52523 pentru NTG, coliformi, staphylococi, cod
71L52524 pentru coliformi, Pseudomonas, streptococi fecali, cod 71L52525
pentru coliformi, staphylococi, drojdii i mucegaiuri,

LOW (+) MEDIUM (++) HIGH (+++)

Fig. 3 - Aprecierea concentraiei microbiene colonii/ml, pe Slide food and water

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Tabelul 1
Indicatorii testului IDEXX Colilert/ Enterolert,

Microorganism Reagent Enzima int Substratul nutritiv


Coliformi Colilert sau - o- nitrophenyl
Colilert 18 galactozidaz galactopyrinozidaz -ONPG
E.coli Colilert sau - 4-methyl umbelliferyl - D-
Colilert 18 glucuronidaz glucuronide - MUG
Enterococi Enterolert -glucozidaz

Tabelul 2
Interpretarea caractere culturale evideniate de Slide food and water

Pseudomona

Staphylococi
Streptococi
Klebsiella
E.coli

Caractere
Mediu

s
morfologice

Aspect rou rou cretere cretere


-
Agar colonie purpuriu purpuriu inhibat inhibat
V.R.B.G. Culoare rou rou cretere cretere
-
mediu purpuriu purpuriu inhibat inhibat
Aspect cretere cretere inhibiie cretere cretere
Agar roz colonie inhibat inhibat parial inhibat inhibat
bengal Culoare
- - - - -
mediu
Aspect
Agar TCC rou rou rou rou rou
colonie
(triptic
Culoare
soia) galben galben galben galben galben
mediu
Aspect cretere cretere cretere cretere
Agar negru
colonie inhibat inhibat inhibat inhibat
Vogel
Culoare
Jonson - - - - galben
mediu
Aspect cretere inhibiie cretere cretere
alb
Agar bil - colonie inhibat parial inhibat inhibat
esculin Culoare
negru
mediu
Aspect cretere cretere cretere cretere
verde
Agar colonie inhibat inhibat inhibat inhibat
Cetrimide Culoare
- - alb - -
mediu

Pentru teste de sanitaie suprafee s-au utilizat metodele clasice admise,


raportate la OMS 976/1998 i metoda rapid bazat pe utilizarea Ultrasnap -uri
ATP Swab interpretate cu Luminat SystemSure II-uri , raportate i interpretate
conform instruciunilor de utilizare ale aparatului.

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Ultrasnap -uri ATP Swab, conin lichidul stabil luciferin/ luciferaz care
induce i catalizeaz reacia de bioluminescen, la contactul cu ATP-ul
molecular, energia regsit la nivelul celulelor bacteriene, a drojdiilor i
mucegaiurilor.
Reacia de bioluminescen are loc prin transformarea energiei chimice n
care sunt implicate legturile pirofosforice ale ATP bacterian n energie
luminoas.

Fig. 4 - Catalizarea n doi pai oxidativi, prin utilizarea ATP ului ,a luciferinului n
oxiluciferin cu producerea unei luminiscene de 560 nm.

REZULTATE I DISCUII

Determinarea calitii microbiologice a apei prin metoda clasic impus


de Stas 3001/91 a fost efectuat pe un numr de 37 probe, din care 9 recoltate n
conformitate cu programul strategic pe 2006, la care s-au analizat 45 de
parametrii i 28 probe recoltate n cadrul programului de autocontrol la care s-au
analizat 78 de parametrii, respectiv determinarea unitilor formatoare de colonii
(UFC/ml), B. colif./100 ml, C. fecali./100 ml, Strept. fecali/100 ml i B. sulfito
red./20 ml
Determinarea calitii microbiologice a apei prin teste rapide de tip
Colilert/ Enterolert 13 probe, la 7 dintre ele solicitarea a fost pentru determinarea
bacteriilor coliforme iar la 5 pentru determinarea enterococilor.
Determinarea calitii microbiologice a apei prin utilizarea Slide food and
water screening s-a realizat pe un numr de 10 probe la care fost investigai
parametrii Bacterii coliforme, Streptococi fecali i Pseudomonas
Consecutiv elaborrii rezultatelor de laborator prin metoda impus de Stas
3001/91, cu raportate la Legea 458/2002, pentru cele 37 de probe se constat c
un numr de 6 probe din cadrul Programului strategic, reprezentnd 16,21% sunt
necorespunztoare - pentru parametrii NTG/ml B.colif./100ml, C.fecali /100ml,
procentul reprezentnd pentru fiecare n parte cte 5,40%.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Pentru analizele din cadrul Programului de autocontrol s-au decelat 7 probe


necorespunztoare, reprezentnd 18,9%, dupa cum urmeaz: cte 5,40% pentru
parametrii NTG/ml B.colif./100ml, i 8,1% pentru parametrul C.fecali /100ml.
Investigaiile rapide privind calitatea microbiologic a apei prin tehnica
impus de modul de lucru al testelor Colilert/ Enterolert au evideniat un numr
de 2 probe necorespunztoare pentru Bacterii coliforme reprezentnd 15,38 %.
Investigaiile rapide privind calitatea microbiologic a apei prin tehnica
impus de modul de lucru cu Slide food and water screening au evideniat un
numr de 2 probe necorespunztoare, pentru parametrul Bacterii coliforme,
reprezentnd 20% i o prob pentru parametrul Streptococi fecali reprezentnd
10%.
Aprecierea strii de igien s-a efectuat prin recoltarea de teste de sanitaie
de pe suprafee de lucru, de pe suprafee utilaje i ustensile, i prin recoltarea de
teste de microaeroflor.
n principal suprafeele de recoltare au fost reprezentate de suprafee de
10/10 cm ale cisternelor de transport lapte tancurilor de rcire, ale vanelor
colectoare de lapte smntnit, conductelor de evacuare lapte smntnit,
conductelor de la instalaiile de pasteurizare, ale crintelor, cuelor, harfelor,
cuitelor, ambalajelor pentru produse finite etc.
Testele de microaeroflor au fost recoltate cu precdere din spaiile
destinate operaiilor tehnologice de presare, maturare i din depozitele produselor
finite.
Determinarea strii de igien s-a efectuat prin recoltarea unui numr de 63
teste de sanitaie din care 52 de pe suprafee utilaje, ustensile, ambalaje i 11 teste
de microaeroflor. Din acestea 25 recoltate n cadrul Programului strategic cu 19
teste de sanitaie suprafee i 6 teste de microaeroflor i alte 38 recoltate n cadrul
Programului de autocontrol, cu 33 teste de sanitaie suprafee i 5 teste de
microaeroflor.
Numrul testelor necorespunztoare este de 32 61,53%, din care 13 teste
reprezentnd 25% pentru parametrul NTGMA, 2 teste pentru enterobacterii
reprezentnd 3,85% pentru enterobacterii, 9 teste reprezentnd 17,3% pentru
Bacterii coliforme i 4 teste pentru parametrul Levuri i fungi reprezentnd
7,69%.
Testele investigate prin utilizarea system Sure II TM ATP relev 2 dintre
ele au afiat o valoare a ATP ntre 11 i 29 ceea ce indic c suprafeele testate
sunt inadecvate i alte 2 au afiat o valoare a ATP peste 30 ceea ce indic c
suprafeele testate sunt murdare.

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Tabelul 3
Investigaii clasice privind calitatea microbiologic a apei utilizate
n procesarea brnzei telemea

Anul B.sulfito
NTG B.colif. C.fecalii. Strept.fecali
2006 Numr red.
/ml /100ml /100ml /100ml
Ianuarie- probe /20ml
Martie C N C N C N C N C N
P.S. 9 7 2 7 2 7 2 9 0 9 0
P.A. 28 20 2 25 3 15 2 8 0 3 0

Tabelul 4
Investigaii rapide privind calitatea microbiologic a apei utilizate
n procesarea brnzei telemea - Colilert / Enterolert

Anul 2006 Numr Coliformi Enterococi


Ianuarie - probe
Martie C N C N
P.A. 13 7 2 5 0

Tabelul 5
Investigaii rapide privind calitatea microbiologic a apei utilizate
n procesarea brnzei telemea Food slide and water 3

Anul 2006 Numr Coliformi Streptococi fecali Pseudomonas


Ianuarie - Martie probe C N C N C N
P.A. 10 8 2 9 1 10 0

Tabelul 6
Investigaii microbiologice privind controlul strii de igien
n unitile de procesare lapte, prin teste de sanitaie

Anul Strept.
2006 Numr NTGMA Enterobacterii Staph.cp. hemolitici B.colif Salm.spp.
Ianuarie probe
C N C N C N C N C N C N
- Martie
P.S. 19 9 4 4 2 4 0 4 0 9 4 4 0
P.A. 26 10 2 3 2 3 0 0 0 9 5 0 0

Tabelul 7
Investigaii microbiologice privind controlul strii de igien
n unitile de procesare lapte, prin teste de microaeroflor

NTGMA D+M
Anul 2006 Numr 3
/600m /300m3
Ianuarie - Martie probe
C N C N
P.S. 6 3 3 4 2
P.A. 5 5 2 5 2

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Tabelul 8
Investigaii microbiologice privind controlul strii de igien
n unitile de procesare lapte, prin utilizarea systemSure II TM ATP

Anul 2006 Numr Valori ATP afiate


Ianuarie - Martie probe < 10 11-29 >30
P.A. 7 3 2 2
** rezultatele necorespunztoare au fost repetate n laborator prin metode clasice

Diagrama 1 - Investigaii privind calitatea apei prin metoda


25 clasica

20

15 15

9 9
7 7 7

2 2 3 3
2 2 2
0 0 0 0
NTG B.colif C.fecali Strept.fecali B.s.red

Program Strategic corespunzatoare Program Strategic necorespunzatoare

Program Autocontrol corespunzatoare Program Autocontrol necorespunzatoare

Diagrama 2 - Investigaii privind calitatea apei prin tehnica


Coililert /Enterolert

Colilert C Colilert N Enterolert C Enterolert N

Colilert C Colilert N Enterolert C Enterolert N

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Diagrama 3 - Investigaii privind calitatea apei prin utilizarea


Slide Food and Water Screening

10
9
8

2
1
0

Coliformi Streptococi fecali Pseudomonas

corespunzatoare necorespunzatoare

Diagrama 4 - Investigaii microbiologice privind controlul


strii de igien n unitile de procesare lapte, prin teste de
10 sanitaie
9 9 9

5
4 4 4 4 4 4
3 3
2 2 2

0 0 0 0 0 0 0 0
NTG B.colif Enterobacterii Staphilococ Streptococ Salmonella
cp. hemolitic spp.

Program Strategic corespunzatoare Program Strategic necorespunzatoare


Program Autocontrol corespunzatoare Program Autocontrol necorespunzatoare

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Diagrama 5 - Investigaii microbiologice privind controlul strii


de igien n unitile de procesare lapte, prin teste de
microaeroflora
5 5 5 5

2 2

NTGMA NTGMA D/M D/M


Program Strategic corespunzatoare Program Strategic necorespunzatoare
Program Autocontrol corespunzatoare Program Autocontrol necorespunzatoare

Diagrama 6 - Investigaii microbiologice privind controlul strii


de igien n unitile de procesare lapte, prin systemSure II ATP

ATP < 10
ATP 11-29
3 ATP >30

2 2

CONCLUZII

Procentul de 26,98% probe de ap necorespunztoare aparin unor uniti


care nu sunt racordate la surs centralizat de ap i se alimenteaz prin reele
proprii fntni prevzute cu hidrofor, lipsite de filtre i amplasamente
necorespunztor n incita unitii.

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Determinarea strii de igien a suprafeelor de lucru, a utilajelor, a minilor


personalului i a microaeroflorei din spaiile de procesare lapte denot existena
unor deficiene majore n ceea ce privete asigurarea igienei pe fluxul tehnologic.
Procentul de 61,53% probe necorespunztoare, asociat cu lipsa implementrii
sistemului HACCP, sau gestionare incorect a punctelor critice n documentele
HACCP n unitile de procesare lapte indic, un management defectuos - lipsa de
fonduri sau consiliere corespunztoare, i o lips a contientizrii procesatorilor
asupra importanei inocuitii produselor, sunt elemente cu repercusiuni nefaste
supra calitii produsului finit i nu n cele din urm asupra supravieuirii pe
,,piaa agroalimentar n condiiile impuse de U.E., pentru libera circulaie a
produselor.

BIBLIOGRAFIE
1. Brzoi D., Sanda Lzrescu, Natalia Maier, Lidia Tulu, R. Korn, Olga Nuc,
Florina Dumitrescu, 1978 - Metode Microbiologice pentru examenul de
laborator al produselor alimentare de origine animal, M.A.I.A., D.S.V.,
L.C.C.A.Bucureti
2. Barzoi D., S. Apostu, 2002 - Microbiologia produselor alimentare, Ed.Risoprint, Cluj
Napoca.
3. Chintescu G., 1980 - ndrumtor pentru tehnologia brnzeturilor, Editura tehnic,
Bucureti,
4. Coman I., Helena-Maria Brhal, Rodica Florescu, 1997 - Decontaminarea vol I,
Ed. Satya, Iai
5. Cpri Rodica, A. Cpri, T. Vintil, 2004 - Aplicaion of the luciferin/luciferase
bioluminescent system in hygiene monitoring of raw milk, Facultatea de chimie
industrial i Protecia mediului, Timioara.
6. Enache T., Paul I., Stnescu V., Popescu O., Iordache I., 1995 - Medicin legal
veterinar. Ed.All, Bucureti.
7. Enache T. Paul I., Stnescu V., Popescu O., Iordache I., 2001 - Tratat complet de
medicin legal veterinar, Ed.All, Bucureti.
8. Mark K., Mitchelland W., Stapp B., 1996 - Field Manual of Water Quality Monitoring
Kendall Hunt Publishess, p.304,
9. Stender H. i colab., 2001 Rapid detection, identification and numeration oe E.Coli
cells in water by chemiluminiscent in situ Hybridization, Apl. Environ.Microbiol.,
67,1,142,
10. Standarde de stat, norme tehnice de calitate i metode de lucru: Stas 3001/1991,SR
ISO 11290/1/2000,
11. SR ISO 4833/1991, SR ISO 5541-1-2 /1994, SR ISO 7251/1996, SR ISO 6888/2002,
SR ISO 6579/2003, SR ISO 7402/1996
12. OMS 976/98,
13. Legea privind calitatea apei potabile 458/2002, Monitorul Oficial al Romniei, partea I
nr. 552/ 29.07.2002, completat cu Legea privind calitatea apei potabile nr.311/
2004, Monitorul Oficial al Romniei 582/ 30. 06.2004.
14. http://www.orgenics.com/files/products/UTI. - Food slide and water,
15. www.idexx.com/water/colilert / enterolert detection of coliforms, E.coli and
enterococci in water.

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NIVELUL UNOR NITROZAMINE VOLATILE


N DIVERSE PRODUSE DIN CARNE I PETE AFUMAT
THE LEVEL OF SOME VOLATILE NITROSAMINES IN VARIOUS MEAT PRODUCTS
AND SMOKED FISH

E.V. INDILAR, M. TOTOLIN, E. INDILAR


U.S.A.M.V. Iai

The research regarding the three nitrosamines NDMA, NDEA,


NPYR in some meat and fish products trough gascromatographic method,
have shown that NDMA and NDEA are present in different quantities in all
seven products that were examined and NPYR was present only in three
sortiments.
The average values of NDMA were: 0,058 g/kg in Choriso
sausages; 0.114 g/kg in Kaiser; 0.120 g/kg in Bnean salami;
0,135 g/kg in Cabanos; 0,160 g/kg in Sibiu salami; 0,193 g/kg in
bacon; 0,331 g/kg in smoked fish.
The average values of NDEA were: 0,043 g/kg in Choriso
salami; 0,045 g/kg in Cabanos; 0,050 g/kg in Bnean salami;
0,051 g/kg in smoked fish; 0,099 g/kg in Sibiu salami; 0,150 g/kg in
Kaiser.
NPYR wasnt found in Sibiu, Choriso, or Bnean salami;
it was present and had average values of: 0,163 g/kg in bacon; 0,260
g/kg in Kaiser and 0,762 g/kg in smoked fish.

Key words: meat products, level, volatile nitrosamines.

Nitrozaminele fac parte din marea grup a nitrozocompuilor. Ele conin


gruparea N-nitrozo, iar radicalii pot fi reprezentai de grupri arilice, alchilice,
aril-alchilice sau heterociclice. Pot fi n stare lichid sau solid i unele din ele
sunt volatile (1, 2, 3, 7, 9). n organism sunt relativ repede biodegradate, dnd
produi reactivi i instabili, dar care constituiesc substane cancerigene. Faza
final a conversiei este un ion de carbeniu nalt electrofil, n cutare de locuri
bogate n electroni cum sunt bazele purinice i piriminidice ale moleculelor de
ARN, ADN, proteinele (2, 3).
Structura lor chimic complex le permite s participe la diverse reacii
chimice de reducere, oxidare, hidroliz i transformare fotochimic (3, 7).
Nitrozaminele se pot forma exogen i endogen (n organism) din precursori
reprezentai de nitrai, nitrii, prioritar amine secundare i aminoacizi (2, 7, 8, 11,
12). Viteza nitrozrii aminelor depinde de natura aminei, concentraia NaNO3 i
NaNO2 i a altor nitrozani i prezena sau absena unor factori catalizatori sau
inhibani (3, 7, 8). ntre factorii catalizatori se nscriu: pH-ul acid (2,5-3,5)

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comparabil cu cel al sucului gastric, temperaturile ridicate, timpul de contact ntre


amine i compuii nitrozani i activitatea metabolic a unor bacterii (2, 7, 10).
ntre factorii inhibani ai formrii NA se nscriu: unii protectori coloidali
(amidonul), acidul galic, acidul ascorbic i ascorbaii, tocoferolii (2, 7, 10).
n alimentele bogate n substane proteice, supuse stocrii, maturrii,
proceselor termice ridicate, fermentrii se pot elibera aminoacizii, care prin
decarboxilare formeaz amine care n prezena nitriilor formeaz NA (7, 10).
In vivo cea mai mare cantitate de NA se formeaz n mediul acid al
stomacului i prin activitatea metabolic a unor bacterii n intestin i cile urinare
(7, 10).
nafara formrii NA n alimente se pot forma i n diverse buturi, ape,
medicamente, pesticide, tutun, cosmetice, colorani, uleiuri, lubrefiani (2, 7).
Rezult c expunerea uman la riscuri NA este dat de diverse modaliti,
iar aciunea lor nociv este foarte divers.
n prezent, pe baza observaiilor la om i animale, i a experimentelor pe
animale, efectele nocive ale NA se pot manifesta prin toxicitate acut,
embriotoxicitate, mutagenitate, carcinogenitate (4, 5, 6, 9, 10, 11, 12, 13, 14, 15).
n lucrare ne-am propus s stabilim nivelul unor NA volatile n unele
produse din carne i pete afumat.

MATERIAL I METOD

Determinrile s-au fcut pe un numr de 6 sortimente de produse din carne


i un sortiment de pete afumat totaliznd 35 probe. S-au determinat nivelurile de
NDMA, NDEA i NPYR ca NA volatile.
Determinrile acestor NA s-au fcut prin metoda gazcromatografic,
respectnd metodologiile adecvate privind extracia, purificarea distilatelor,
dozarea NA prin metoda gazcromatologic, confirmarea rezultatelor prin
spectrometrie de mas i interpretarea cromatogramelor.

REZULTATE I DISCUII

Rezultatele obinute sunt redate n tabelul nr. 1 i diagramele 1, 2, 3, 4.


n cele 6 sortimente de produse din carne i sortimentul de pete afumat s-a
pus n eviden dimetilnitrozamina (NDMA) n urmtoarele cantiti (diagrama
1):
- 0,134-0,263 g/kg la bacon i o valoare medie de 0,193 g/kg;
- 0,102-0,218 g/kg la salamul de Sibiu i media de 0,160 g/kg;
- 0,104-0,160 g/kg la crnai Cabanos, iar media de 0,135 g/kg;
- 0,100-0,150 g/kg la salamul Bnean i medie de 0,120 g/kg;
- 0,092-0,147 g/kg la Kaiser cu valoare medie de 0,114 g/kg;
- 0,040-0,070 g/kg la salamul Choriso cu media de 0,058 g/kg;
- 0,200-0,512 g/kg la petele afumat i media de 0,331 g/kg.

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Tabelul 1
Valori medii ale coninutului de NDMA, NDEA i NPYR n unele produse din carne
i pete afumat

n = 5 : NDMA n = 5 : NDEA n = 5 : NPYR


Produsul din
micrograme/kg micrograme/kg micrograme/kg
carne sau pete
Limite Media Limite Media Limite Media
Bacon 0,134-0,263 0,193 0,041-0,060 0,052 0,104-0,206 0,163
Cabanos 0,104-0,160 0,135 0,030-0,058 0,045 urme urme
Kaiser 0,092-0,147 0,114 0,113-0,175 0,155 0,200-0,337 0,260
Salam Bnean 0,100-0,150 0,120 0,040-0,060 0,050 0 0
Salam Choriso 0,040-0,070 0,058 0,030-0,060 0,043 0 0
Salam de Sibiu 0,102-0,218 0,160 0,088-0,108 0,099 0 0
Pete afumat 0,200-0,512 0,331 0,040-0,060 0,051 0,559-1,002 0,762
Limite/Media 0,040-0,512 0,159 0,030-0,175 0,071 0-1,002 0,169

0,35 0,331

0,3

0,25
0,193
0,2
g/kg 0,16
NDMA 0,135
0,15 0,12
0,114
0,1
0,058
0,05

0
S. Choriso Kaiser S. Bnean C. Cabanos S. Sibiu Bacon Pete
afumat

Diagrama 1. - Valori medii ale coninutului de NDMA n unele produse din carne i pete
afumat (g/kg)

Nivelul de dietilnitrozamin (NDEA) n produsele de carne i pete a fost


relativ mai mic ca cel al NDMA, cu excepia sortimentului Kaiser. NDEA a avut
urmtoarele valori limit i medii (diagrama nr. 2):
- 0,030-0,060 g/kg i media de 0,043 g/kg la salamul Choriso;
- 0,030-0,058 g/kg cu media de 0,045 g/kg la crnai Cabanos;
- 0,040-0,060 g/kg i valoare medie de 0,050 g/kg la salamul Bnean;
- 0,040-0,060 g/kg i valoare medie de 0,051 g/kg la petele afumat;
- 0,041-0,060 g/kg i media de 0,052 g/kg la bacon;
- 0,088-0,108 g/kg i media de 0,099 g/kg la salamul de Sibiu;
- 0,113-0,175 g/kg i media de 0,155 g/kg la Kaiser.

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0,2

0,155
0,15

0,099
g/kg
0,1
NDEA

0,05 0,051 0,052


0,043 0,045
0,05

0
S. Choriso C. Cabanos S. Bnean Pete Bacon S. Sibiu Kaiser
afumat

Diagrama 2. Valori medii ale coninutului de NDEA n unele produse de origine animal

1
0,9
0,762
0,8
0,7
0,6
NPYR
g/kg

0,5
0,4
0,26
0,3
0,163
0,2
0,1 0 0 0
0
0
C. Cabanos S. Bnean S. Choriso S. Sibiu Bacon Kaiser Pete
afumat

Diagrama 3. Valori medii ale coninutului de NPYR n unele produse de origine animal

- 0,088-0,108 g/kg i media de 0,099 g/kg la salamul de Sibiu;


- 0,113-0,175 g/kg i media de 0,155 g/kg la sortimentul Kaiser.
NPYR, cea de a treia NA volatil cercetat n produsele din carne i pete
(diagrama 3) a fost:
- absent n salamul Bnean, salamul Choriso i salamul de Sibiu;
- sub form de urme n crnaii Cabanos;

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- prezent cu valori medii de - 0,163 g/kg (0,104-0,206 g/kg) la Bacon;


0,260 g/kg (0,200-0,337 g/kg) la Kaiser; 0,762 g/kg (0,559-1,002 g/kg) la
petele afumat.
Cumulate nivelurile celor trei NA (NDMA, NDEA i NPYR) din produsele
cercetate, s-a putut constata urmtoarele valori medii (diagrama nr. 4): 1,114
g/kg la petele afumat; 0,529 g/kg la Kaiser; 0,408 g/kg la bacon; 0,259 g/kg
la salamul de Sibiu; 0,180 g/kg la Cabanos; 0,170 g/kg la salamul bnean i
0,101 g/kg la salamul Choriso.

1,2 1,114
1,1
1
0,9
0,8
0,7
g/kg 0,6 0,529
0,5 0,408
0,4
0,259
0,3 0,18 0,17
0,2 0,101
0,1
0
Pete Kaiser Bacon S. Sibiu C. S. S. Choriso
afumat Cabanos Bnean

Diagrama 4. Valori medii cumulate de NDMA, NDEA i NPYR n unele produse de origine
animal (g/kg)

CONCLUZII

Investigaiile privind nivelul de NDMA, NDEA i NPYR n unele produse


din carne i pete afumat au evideniat:
1. Prezena NDMA la toate cele 7 sortimente cercetate la nivel mediu de
0,058 g/kg la salam Choriso i de 0,331 g/kg la petele afumat.
2. NDEA s-a identificat la fel n toate cele 7 sortimente, dar n cantiti
inferioare fa de NDMA cu excepia produsului Kaiser.
3. NPYR a avut o valoare medie de 0,762 g/kg la petele afumat, 0,260
g/kg la Kaiser, 0,163 g/kg la bacon, ca urme n Cabanos i absent n salamurile
de durat.

BIBLIOGRAFIE

1. Andrade R., Reyes G.R., Rath Susanne, 2005 A method for the determination of
volatile N-nitrosamines in food by HS-SPME-GC-TEA. Food Chemistry, 91, 1,
173-179.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

2. Hura Carmen, 1997 Poluarea chimic a alimentelor i sntatea. Ed. Socom


Hermes, Iai.
3. Hendrickson B.J., Cram J.D., Hamonand S.G., 1976 Chimie organic. Ed. t. i
Enciclopedic, Bucureti.
4. Jgerstad Margaretha, Skog K., 2005 Genotoxicity of heat procesed foods.
Mutat. Reserche, 574, 1-2, 156-172.
5. Montesano R., Bartsch H., 1976 Mutagenic and carcinogenic N-nitrocompouds:
Posibil environmental hazard. Mutat. Res., 32, 179-227.
6. Mirvish S.S., 1995 Role of N-nitroso compounds and nitrosation in etiologi of
cancer gastric, oesophageal, nasopharyngeal and bladder cancer and
contribution to cancer of known exposure to NOC. Cancer Lettres 93, 17-25.
7. Popa G., Segal B., Dumitrache S., Segal Rodica, Apostol C., Teodoru V., 1986
Toxicologia produselor alimentare. Ed. Academiei Romne, Bucureti.
8. Stnescu V., Stnescu Undina, Ciocrlie Nicoleta, 1996 Semnificaia igienico-
sanitar a nitrozaminelor n produsele alimentare. Rev. Rom. de Med. Vet., 6, 3,
347-359.
9. indilar E., Bondoc I., 1994 Efectul toxic i cancerigen al nitrozaminelor din
alimente. Tezele Conf. jubiliare 20 de ani de nvmnt vetrinar n R. Moldova
105.
10. indilar E.V., 2005 Contribuii la studiul reziduurilor de nitrai, nitrii i nitrosamine
n unele produse de origine animal. Tez de doctorat, F.M.V. Iai.
11. Ukava-Ishikava S., Seki M., Mochizuki M., 1999 Mutagenity of potasium
Alkanediazotates in Chinese Hamster V79 Cells and their Alkilanting Activity.
Biol. Pharm. Bull., 22, 6, 577-581.
12. Yurchenko S., Mlder U., 2005 Volatile N-nitrosamines in various fish products.
Food Chemistry, Abstract, Article in Press.
13. Environmental Protection Agency-IRIS, 2005 N-nitrosodimethylamine.
http://www.epa.gov/subst/0045.htm
14. Environmental Protection Agency-IRIS, 2005 N-nitrosodiethylamine.
http://www.epa.gov
15. Environmental Protection Agency-IRIS, 2005 N-nitrosopyrolidine.
http://www.epa.gov/iris/subst/0081.htm

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NIVELUL UNOR NITROZAMINE VOLATILE


N UNELE SORTIMENTE DE BRNZETURI
THE LEVEL OF SOME VOLATILE NITROSAMINES IN VARIOUS CHEESES

E.V. INDILAR, M. TOTOLIN, E. INDILAR


U.S.A.M.V. Iai

Trough gascromatographic method we are examined the level of


NDMA, NDEA and NPYR in varius cheese.
In cheese, the following aspects regarding nitrosamines were shown:
- Neither NDMA, NDEA and NPYR were found in Balkan
cheese;
- in Ceddar cheese there were found only traces of the three
nitrosamines;
- only NDMA was found in Schweitzer cheese, with the
average value of 0,069 g/kg;
- the three nitrosamines were present in smoked Cacaval
cheese as follows: 0,111 g/kg for NDMA; 0,063 g/kg for
NDEA; 0,025 g/kg for NPYR.

Key words: cheese, nitrosamines, level

Nitrozaminele sunt compui chimici toxici de risc cancerigen indirect,


clasate n categoria toxicelor dup bioactivare (metabolizare). n laptele i
brnzeturile proaspete, iaurt, unt, smntn sunt absente sau sub form de urme.
n laptele pasteurizat s-au gsit NA n cantiti de 0,05-0,17 g/kg, iar n laptele
praf n limite de 0-4 g/kg i o medie de 1,9 g/kg (4, 6).
n brnzeturi, n special n cele maturate s-a stabilit niveluri de pn la 20
g/kg (6).
Se consider c nivelul de nitrozare din aceste brnzeturi este dat de
procesul de maturare i intervenia unor microorganisme. Astfel, Penicillium
camemberti i ali micei favorizeaz formarea de NA. n brnza Ceddar i
Camembert s-au evideniat niveluri cuprinse ntre 0,9-1,7 g/kg. n brnzeturile
de tip Roquefort s-au identificat NDMA i NDEA, iar brnza Schweitzer
NDMA, NDEA i NPIP, dar la niveluri relativ mici la unele probe (4, 6).
n unele brnzeturi daneze, NA la niveluri de 0,1-0,3 g/kg s-a stabilit la
26% din 156 de probe cercetate (6, 7).
Aciunea NA asupra oamenilor i animalelor se manifest extrem de diferit
n raport de doz, structur chimic inducnd de la oc toxic la mutagenitate,
carcinogenitate i embriotoxicitate (1, 2, 5, 8, 9, 10).
Aciunea carcinogen a NA poate fi explicat prin ipoteza alchilrii. n
organism NA sunt supuse unui proces de activare biologic printr-o reacie de

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alchilare cu formare de alfa hidroxinitrozamin, care n final conduce la un agent


alchilant-diazoalcan, care va aciona la nivelul sediilor nucleofile ale ADN i
ARN (1, 3, 6, 9).
n lucrare s-a urmrit stabilirea nivelului de NDMA, NDEA i NPYR la
patru sortimente de brnzeturi: Ceddar, Schweitzer, Telemea i Cacaval afumat.

MATERIAL I METOD

Pe 20 probe de diverse sortimente de brnzeturi s-au determinat prin


metoda gazcromatografic trei NA volatile: NDMA, NDEA i NPYR.
Determinrile s-au fcut respectnd cerinele complexe impuse de
metodologia extraciei, purificarea extractelor, etalonarea corect, dozarea NA
gazcromatografic, confirmarea rezultatelor prin spectrometrie de mas i
interpretarea cromatogramelor.

REZULTATE I DISCUII

Rezultatele obinute sunt redate n tabelul nr. 1 i diagrama 1.


Tabelul 1
Valori ale coninutului de NDMA, NDEA n unele brnzeturi

n=5: n=5: n=5:


Produsul
NDMA g/kg NDEA g/kg NPYR g/kg
din lapte
Limite Media Limite Media Limite Media
Cedar urme urme urme urme urme urme
vaier 0,055-0,075 0,069 0 0 0 0
Telemea 0 0 0 0 0 0
Cacaval
0,100-0,125 0,110 0,054-0,075 0,063 0,020-0,037 0,025
afumat
Media 0-0,125 0,044 0-0,075 0,016 0-0,0,37 0,006

Datele obinute au evideniat urmtoarele valori ale nitrozaminelor n cele


patru sortimente de brnzeturi cercetate:
- coninut 0 g/kg de NDMA, NDEA i NPYR n brnza telemea;
- urme de NDMA, NDEA i NPYR n brnza Ceddar;
- coninut ntre 0,055-0,075 g/kg i o medie de 0,069 g/kg de NDMA
la brnza vaier i coninut 0 g/kg de NDEA i NPYR;

- coninut ntre 0,100-0,125 g/kg ( x = 0,110 g/kg) de NDMA, ntre

0,054-0,075 g/kg( x = 0,063 g/kg) de NDEA i ntre 0,020-0,037

( x = 0,025 g/kg) de NPYR n cacavalul afumat.

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Datele au evideniat cteva aspecte interesante:


- absena celor trei NA cercetate n brnza telemea ca produs lactat
apreciat de consumatori;
- prezena numai a NDMA n brnza Schweitzer cu un nivel mediu
0,069 g/kg;
- prezena NDMA, NDEA i NPYR n cacavalul afumat.
Aceste valori sunt sub LMA prevzut n normative.

0,11
0,11
0,1
0,09
0,08 0,069
0,063
0,07
g/kg 0,06
0,05
0,04
0,025
0,03
0,02
0,01 0 0 0 0 0 0 0 0
0
NDMA NDEA NPYR

B. Cedar B. telem ea B. vaier Cacaval afum at

Diagrama 1. Valori medii ale coninutului de NDMA, NDEA i NPYR n unele brnzeturi

CONCLUZII

Investigaiile privind stabilirea nivelurilor de NA volatile din unele


sortimente de brnzeturi ne-au condus la stabilirea urmtoarelor concluzii:
1. La toate sortimentele de brnzeturi, nivelul de NDMA, NDEA i NPYR
s-a situat sub LMA normat.
2. n brnza telemea nu s-au evideniat NDMA, NDEA i NPYR.
3. Prezena de urme de NDMA, NDEA i NPYR n brnza Ceddar.
4. Prezena numai de NDMA cu valoare medie de 0,069 g/kg n brnza
schweitzer.
5. Prezena NA volatile n cacavalul afumat cu nivel mediu de 0,110 g/kg
de NDMA; 0,063 g/kg de NDEA i 0,025 g/kg de NPYR.

BIBLIOGRAFIE

1. Bartsch H., Ohsihima H., Shuker D.E., Pignatelli B., Calmels S., 1990 Exposure
of humans to endogenous N-nitroso compounds implications in cancer etiology.
Mutat. Res., 238, 255-267.
2. Bruning-Fann C.S., Kaneene J.B., 1993 The effects of nitrate, nitrite and N-nitroso
compounds on human health: a rewiew. Vet. Human Toxicology, 35, 521-538.

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3. Corne Jaqueline, 1992 Estimations de la concentration des nitrites, nitrates et


nitrosamine dans quelques denre en France. Science des aliments, 12, 156-
197.
4. Hura Carmen, 1997 Poluarea chimic a alimentelor i sntatea. Ed. Cermi, Iai.
5. Mavelle T., 1991 Dosage de composs N-nitross. In: Technique danalyse et
control dans les industries alimentaires. Technique et Documentation, Paris.
6. Popa G., Segal B., Dumitrache S., Segal Rodica, Apostol C., Teodoru V., 1986
Toxicologia produselor alimentare. Ed. Academiei Romne, Bucureti.
7. Renner E., 1983 Milck and Dairy Products in human nutrition W. Gmb. H,
Volkswirtschaft Pichar Verlag, Mnchen.
8. indilar E.V., 2005 Contribuii la studiul reziduurilor de nitrai, nitrii i nitrosamine
n unele produse de origine animal. Tez de doctorat, F.M.V. Iai.
9. Vermeer T.M. Ingrid, Pachen Danielle, Dallinga W.J., Kleijas C.S.J., Maanen
M.S.J., 1998 Volatile N-nitrosamine Formation after Intake of Nitrat at the ADI
Level in Combination with an Amine-rich Diet. Environ Heaakth Perspect., 106,
459-463.

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APRECIEREA NIVELULUI DE NITRAI N CARNEA


PROASPT DE BOVIN, OVIN, SUIN I PETI
THE ESTIMATION OF NITRATE LEVEL IN COW, EWE, PIG AND FISH RAW MEAT

E.V. INDILAR, E. INDILAR, I. BONDOC


U.S.A.M.V. Iai

We are examined 106 samples of raw meat and 20 samples of raw


fish regarding the nitrate level trough spectrocolorimetric method.
The results regarding the residual level of nitrates in raw meat and

raw raw fish has the next values in ppm NaNO3: 0-24,00 ( x =8.72) for cow

meat; 0-19,00 ( x =9,50) for ewe meat; 6,00-15,00 ( x =10,59) for fish meat;

0-48,00 ( x =17,25) for pig meat.

Key words: nitrates, level, fresh meat, fish.

Nitraii din sol provenii din biodegradare gunoiului i/sau din utilizarea
substanelor nitrice ca fertilizani intr n lanul trofic alimentar sol plant
animale om, produse animale om (9, 11).
Principalele surse de nitrai pentru om sunt reprezentate de: unele vegetale
care cumuleaz n anumite condiii nitraii n rdcini, frunze, tulpini, etc.; apa cu
un coninut de peste 45 mg NO-3/l (1, 2, 7).
Produsele animale procesate i carnea proaspt constituie surse secundare
de nitrai. Dei animalele rumegtoare pot ingera cantiti relativ mari de nitrai
odat cu furajele i apa, n carnea lor dup sacrificare i n laptele lor se regsesc
cantiti relativ mici de nitrai (11, 12).
Rumegtoarele folosesc azotaii ca materie prim pentru sinteza
substanelor azotoase cu ajutorul simbionilor rumenali (11). La monogastrice ,
azotaii sunt absorbii n intestinul subire, trec n circulaia sanguin i sunt
excretai prin rinichi (7, 11).
n carnea procesat n produse din carne la care srarea se face din amestec
de NaCl, NaNO3 i NaNO2 nivelul rezidual al nitrailor rareori depete LMA
(11, 12).
Nitraii sunt redui enzimatic n nitrii att exogen ct i endogen:
- pe baza nitratreductazelor proprii alimentelor i a celor bacteriene din
alimente;
- reducere enzimatic pe baza nitratreductazelor bacteriene din tractusul
digestiv buco-gastro-intestinal (3, 4, 7, 11).

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Nitraii per se au aciune toxic relativ redus (9, 11). Convertii n nitrii
induc efecte hemoglobinizante mai ales la copii (5, 7). Convertii n nitrii ei pot
produce nitrozamine (7, 8, 10).
Att nitraii ct i nitriii perturb la nivelul tubului digestiv, procesele
metabolice ale vitaminelor B1, B6, A, betacarotenului, reduc absorbia proteinelor
i lipidelor din raii, scad activitatea acetilcolinesterazelor, inhib captarea iodului
de ctre tiroid, induc depresia sistemului imunitar i perturb fixarea fierului n
hematii (6, 8, 10, 11).
n lucrare ne-am propus s stabilim nivelul de nitrai din carnea proaspt
de vit, oaie, porc i pete proaspt neprelucrat.

MATERIAL I METOD

Pe 106 probe de carne proaspt de bovine, ovine i suine i 20 probe de


pete proaspt s-a determinat nivelul nitrailor.
Pentru determinare s-a folosit metoda colorimetric cu M-xilenol n care
nitraii i nitriii s-au stabilit ca azot total, iar nitriii s-au determinat prin metoda
Griess.

REZULTATE I DISCUII

Sinteza investigaiilor este redat n tabelul nr. 1 i diagrama 1.


Tabelul 1
Valori ale nivelului de nitrai n carnea proaspt a unor animale de mcelrie i
pete neprelucrat

Specia de la care mg NaNO3/kg


Nr. probe Obs.
provine carnea Intervale Media
Bovine 41 0-24,00 8,72 -
Ovine 25 0-19,00 9,50 -
Suine 40 0-48,60 17,25 -
Pete afumat 20 6,00-15,00 10,69 -
Total/Media 126 0-48,60 11,90

Analiza datelor arat c nivelul nitrailor reziduali a variat n limite relativ


largi n carnea celor trei specii cercetate.
La cele 41 probe de carne de bovin, nitraii au avut valori limit ntre 0
ppm NaNO3 i 24,00 ppm NaNO3, iar valoarea medie de 8,72 ppm NaNO3.
La cele 25 probe de carne de ovin nitraii au avut intervale cuprinse ntre
0-19,00 ppm NaNO3, cu valoare medie de 9,50 ppm NaNO3.
La carnea de suine, constituit din 40 probe, nitraii au avut limitele
cuprinse ntre 0-48,60 ppm NaNO3 i o medie de 17,25 ppm NaNO3.
Nivelul nitrailor la cele 20 probe de pete proaspt a avut limite cuprinse
ntre 6,00 i 15,00 ppm NaNO3 i media de 10,69 ppm NaNO3.

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La carnea mamiferelor bovine, ovine i suine s-a observat absena


nitrailor la 11 probe reprezentnd 10,37%. La carnea de bovine 6 probe (14,63%)
din 41 nu au coninut nitrai. n carnea de ovine nitraii au avut valoarea 0 ppm la
8,00% din probe, iar carnea de porc a fost liber de nitrai la 7,5% din probe.
n carnea petilor proaspei provenii din apele dulcicole din zona Iai,
coninutul nitrailor a avut limite de variaii ntre 6,00 i 15,00 ppm NaNO3 i o
medie de 10,69 ppm NaNO3.

17,25
18

16

14
11,9
12
10,69
9,5
10 8,72
mg/kg
8

0
Carne bovine Carne ovine Carne suine Carne pete Media

Carne bovine Carne ovine Carnea suine Carne pete Media

Diagrama 1. - Valori medii a reziduurilor de nitrai n carnea proaspt i pete neprelucrat

CONCLUZII

n carnea proaspt de bovin i ovin coninutul nitrailor reziduali au avut


valori medii sub 10 ppm NaNO3, dar a fost aproape dublu n carnea de porc
(17,25 ppm NaNO3).
Au fost libere de nitrai 11 probe de carne cercetat reprezentnd 10,37%
din probe.
Nitraii reziduali n carnea petilor proaspei au aprut uor crescui fa de
carnea de bovine i ovine, avnd valoarea medie de 10,93 ppm NaNO3.

BIBLIOGRAFIE

1. Banu C., Preda N., Vasu S., 1982 Produsele alimentare i inocuitatea lor. Ed.
Tehnic, Bucureti.
2. Bara C., Laslo C., 1997 Elemente de ecotoxicologie i protecia mediului
nconjurtor. Ed. Universitii Oradea.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

3. Bruning-Fann C.S., Kaneene J.B., 1993 The effects of nitrate, nitrite and N-nitroso
compounds on human health: a rewiew. Vet. Human Toxicology, 35, 521-538.
4. Cotru M., 1991 Toxicologie. Ed. Did. i Ped. Bucureti.
5. Environmental Protection Agency (E.P.A.U.S), 2005 Nitrates
http://www.epa.gov/iris/subst/0076.htm
6. FRASER P., 1985 Nitrates: epidemiologic evidence. IARC Sci. Publ., 65, 183-185.
7. Hura Carmen, 1997 Poluarea chimic a alimentelor i sntatea. Ed. Socom
Hermes, Iai.
8. Hura Carmen, 2001 Contaminani chimici n produsele alimentare 1980-2000. Ed.
Cermi, Iai.
9. Mitrnescu Elena, Savu C., 1998 Din riscurile polurii mediului i alimentelor. Ed.
MAST, Bucureti.
10. Pignatelli B., 1994 Nitrates, source indirectes de composs N-nitross (CNO).
Risque pour la sant houmain. Soc. Fr. Sant Publique, 1, 62-78.
11. Popa G., Segal B., Dumitrache S., Segal Rodica, Apostol C., Teodoru V., 1986
Toxicologia produselor alimentare. Ed. Academiei Romne, Bucureti.
12. indilar E.V., 2005 Contribuii la studiul reziduurilor de nitrai, nitrii i nitrosamine
n unele produse de origine animal. Tez de doctorat, F.M.V. Iai.

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EVALUAREA NIVELULUI DE NITRII


N UNELE PRODUSE DIN CARNE
THE ESTIMATION OF THE NITRITE LEVEL IN SOME MEAT PRODUCTS

E.V. INDILAR, E. INDILAR


U.S.A.M.V. Iai

We are investigated 1029 samples of meat products regarding the


level of residual nitrites through spectrocolorimetric method.
In all samples examined we are observed wide limits of nitrite level
(4 ppm NaNO2 of smoked products and 121 ppm NaNO2 of fresh salami).
We are established over MRL the level of residual nitrites in:
29,38% for fresh sausages; 8,99% for salami; 6,00% for length sausages;
4,24% for semismoked sausages and 3,09% for smoked products.
For all meat products the residual nitrites over MRL was been in
9,23% of samples.

Key words: meat products, level, residuale nitrites.

Nitriii se folosesc ca adjuvani n produsele din carne n scop tehnologic i


antibacterian (1, 2, 4, 5, 8, 12).
Sub raport tehnologic, mai ales n procesrile care reclam tratamente
termice ale produselor, nitriii alturi de sare induc cteva efecte benefice privind
aroma, gustul i culoarea prin apariia pigmenilor roii de Mi i Hb (1, 2, 4, 5, 8).
Aroma i gustul acestor produse sunt determinate de procesele biochimice
din carne i nitrii (1, 2).
Apariia pigmenilor roii azooximiocromogen i azooxihemocromogen
d aspect plcut produselor asigurndu-le o culoare roie (1, 8, 12). Nitriii sunt
redui n oxid de azot de ctre nitritreductaz, care la cald se combin cu
mioglobina i hemeglobina dnd pigmenii roii (1, 2, 12).
Efectul antibacterian al nitriilor, dei nu n totalitate explicat se manifest
att asupra florei saprofite, dar cu precdere asupra florei patogene anaerobe: Cl.
botulinum, Cl. perfringens (2, 4, 12).
Efectul cid al nitriilor asupra clostridiilor trebuie privit ca o interaciune
ntre temperatur, concentraia NaCl i a NaNO2, care induc scderea Aw-ului,
pH, formarea de substane inhibitoare a clostridiilor de tip Efect Perigo, sau alte
substane cu rol antimicrobian (1, 2, 4, 8).
ntruct nitriii utilizai ca aditivi n produsele din carne s-au dovedit a fi
precursorii formrii de nitrozamine cu efecte cancerigene prin biodegradare, s-au
impus msuri restrictive asupra cantitilor folosite n inducerea efectelor scontate
sub raport tehnologic i asigurarea stabilitii i securitii produselor (1, 2, 3, 4).

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La fel s-a recomandat de ctre organismele internaionale (OMS, FAO) ca


pe lng reducerea cantitilor de nitrii adugai produselor din carne s se
foloseasc concomitent alturi de sare i nitrii, acidul ascorbic sau srurile lui
care reduc nitriii n oxid de azot, coboar potenialul redox, protejeaz
nitrozocromogenii formai i inhib formarea nitrozaminelor (6, 11, 12, 13, 14).
Fiecare ar, innd cont de recomandrile mai sus artate i-a stabilit
cantitile de nitrii adugai n produse n aa fel ca nivelul nitriilor reziduali din
produse s fie ct mai mic, care pentru ara noastr apare ca LMA de 70 ppm
NaNO2. Aceast cerin s-a impus pentru a se respecta recomandarea FAO-OMS
ca ADI pentru NaNO2/kilocorp/zi s fie maximum 0,2 mg i pentru substana
activ ca NO-2/kilocorp/zi de maximum 0,13 mg.
ntruct nitriiiau un potenial toxic ridicat determinnd
methemoglobinemie i sunt precursorii formrii de nitrozamine cancerigene prin
combinarea lor cu aminele secundare prioritar, fiecare ar are obligaia s
monitorizeze i s supravegheze permanent prin laboratoare agreate nivelul
reziduurilor de nitrii din produse (7, 9, 10, 15, 16).
n lucrare s-a urmrit nivelul rezidual al nitriilor la unele sortimente de
mezeluri i afumturi comercializate pe piaa municipiului Iai n perioada a mai
muli ani (2001-2004).

MATERIAL I METOD

Prin tehnica spectrocolorimetric, folosind metoda Greiss s-au determinat


nitriii reziduali din 177 probe de diverse sortimente de prospturi, 189 probe de
diferite sortimente de crnai, 259 porbe de diferite sortimente de salamuri
semiafumate, 354 probe de diverse afumturi i 50 probe de diferite sortimente
de salamuri de durat.

REZULTATE I DISCUII

n tabelul nr. 1 i figura nr. 1 se red nivelul rezidual al nitriilor din patru
sortimente de prospturi indigene i trei sortimente de prospturi din import.
Datele obinute la sortimentele indigene au evideniat faptul c nitriii
reziduali prezint limite de variaii relativ largi pentru fiecare sortiment cercetat i
la dou sortimente s-au observat depiri ale LMA, adic la 7,69% din probele de
crenwurti i la 2% din probele indigene de parizer clasic.
La cele trei sortimente de prospturi din import valorile limit dei au
prezentat variaii relativ strnse, au avut depiri ale LMA la 93,33% din probele
de parizer de curcan, la 95,00% din probele de parizer de porc i la 93,33% din
probele de parizer de pui.

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Tabelul 1
Valori ale reziduurilor de nitrii n cteva sortimente de prospturi
n perioada 2001-2004 i procentul depirii LMA

Frecvena
Nr. Nr. mg NaNO2/kg depirii
Sortimentul
crt. probe LMA
Limite Media Abs. Rel.
A. Prospturi indigene
1 Crenwurti 52 10,20-121,00 29,64 4 7,69
2 Parizer clasic 50 10,00-72,80 26,98 1 2,00
3 Parizer pui 10 29,00-45,00 36,60 0 0
4 Polonez 15 10,80-46,00 26,45 0 0
Total/Medie 127 10,00-121,00 28,76 5 3,93
B. Prospturi import
1 Parizer curcan 15 70,00-82,00 74,53 14 93,33
2 Parizer porc 20 70,00-90,00 83,10 19 95,00
3 Parizer pui 15 70,00-94,00 85,73 14 93,33
Total/Media 50 70,00-94,00 81,32 47 94,00
Total gen./Media 177 10,00-121,00 43,61 52 29,38

93,33 95 93,33
100 94
90 Crenwurti
80
Parizer
70
P. pui
60
% 50 Polonez

40 Media
30
P. curcan
20
7,69
10 2 3,93
00
0
sortimente sortimente
indigene import

Figura 1. - Procentul depirii LMA de nitrii n unele sortimente de prospturi indigene


i din import

Pentru cele 189 probe de crnai, aparinnd la nou sortimente, rezultatele


sunt redate n tabelul nr. 2 i figura nr. 2.
Analiza datelor a evideniat c nitriii reziduali prezint largi limite de
variaii la circa 78% din cele nou sortimente cercetate cu valori cuprinse ntre 15
ppm NaNO2 la crnaii de porc i 111 ppm NaNO2 la crnaii Trandafir.

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Cinci sortimente din cele nou cercetate (circa 55%) au prezentat depiri
ale LMA de 70 ppm NaNO2. Aceste depiri ale LMA s-au ntlnit n procent de
3,70% la crnaii de Cluj; 14,28% la crnaii Cabanos; 14,28 la crnaii Trandafir;
20,00% la crnaii de porc i 20,00% la crnaii populari. La aceast categorie de
produse s-au nregistrat depiri ale LMA la 8,99% din probe.
Tabelul 2
Valori ale reziduurilor de nitrii din unele sortimente de crnai
i procentul depirii LMA n perioada 2001-2004

Frecvena
Nr. Nr. mg NaNO2/kg depirii
Sortimentul
crt. probe LMA
Limite Media Abs. Rel.
1 Crnai Cabanos 42 18,00-88,00 49,37 6 14,28
2 Crnai de cas 32 29,00-52,00 38,71 0 0
3 Crnai Cluj 27 23,00-75,00 44,71 1 3,70
4 Crnai Mnzteti 10 36,00-53,00 43,60 0 0
5 Crnai Muntenia 10 38,00-50,00 46,10 0 0
6 Crnai Olteneti 10 35,00-42,00 37,20 0 0
7 Crnai popularii 5 31,00-101,00 54,80 1 20,00
8 Crnai de porc 25 15,00-93,00 49,16 5 20,00
9 Crnai Trandafir 28 20,00-111,00 45,52 4 14,28
Total/Medie 189 15,00-111,00 45,33 17 8,99

70

60

50

40
%
30
20 20
14,28 14,28
20
8,99
10 3,7

0
C. Cluj C. Cabanos C. Trandafir C. populari C. porc Media

Figura 2. - Procentul depirii LMA de nitrii n unele


sortimente de crnai

Rezultatele privind nivelul reziduurilor de nitrii n unele sortimente de


salamuri semiafumate sunt redate n tabelul nr. 3 i figura nr. 3.
La cele 259 de probe de salamuri semiafumate, aparinnd la 14 sortimente
diferite, valoarea medie a nitriilor reziduali a fost de 40,57 ppm NaNO2. i la

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aceast categorie de produse s-a remarcat variaii largi ale valorilor limit
cuprinse ntre 4 ppm NaNO2 la salamul de porc i 104 ppm NaNO2 la salamul de
vit+porc.
Tabelul 3
Valori ale reziduurilor de nitrii n unele sortimente de salamuri semiafumate
cu evidenierea depirii LMA

Frecvena
Nr. Nr. mg NaNO2/kg
Sortimentul depirii LMA
crt. probe
Limite Media Abs. Rel.
10 Salam Bicaz 10 32,80-40,50 39,03 0 0
11 Salam Brate 8 30,00-65,00 50,50 0 0
12 Salam Bucureti 50 25,00-53,00 41,58 0 0
13 Salam Debrein 10 35,50-72,10 41,76 1 10,00
14 Salam Dumbrava 18 36,00-42,00 38,57 0 0
15 Salam Franuzesc 5 41,00-46,00 42,80 0 0
16 Salam Italian 40 11,00-55,00 39,71 0 0
17 Salam Mistreul 10 38,80-40,50 40,40 0 0
18 Salam Picant 10 33,00-41,00 38,30 0 0
19 Salam de porc 12 4,00-68,00 34,42 0 0
20 Salam de Vara 56 7,00-102,00 40,11 4 7,14
21 Salam Voinea 10 35,50-47,70 38,00 0 0
22 Salam vit+porc 8 14,00-104,00 56,87 2 25,00
23 Salam Vntoresc 12 7,30-79,00 36,53 1 8,33
Total/Medie 259 4,00-104,00 42,57 8 3,09

70

60

50

40
%
30 25

20
7,14 8,33 10
10 3,09

0
S. Var S. Vntoresc S. Debrein S. vit+porc Total depiri
LMA

Figura 3. - Procentul depirii LMA de nitrii n unele


sortimente de salamuri semiafumate

Depiri ale LMA de nitrii reziduali s-au nregistrat pe sortimente n


procent de: 7,14% la salamurile de var; 8,33% la salamul Vntoresc; 10,00% la

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salamul Debrein i 25,00% la salamul vit+porc. Pentru toat gama sortimental


de salamuri semiafumate depirea LMA s-a ntlnit la 3,09% din probele
cercetate.
La salamurile de durat, media nitriilor reziduali a fost de 56,96 mg
NaNO2/kg, dar i la aceast categorie de produse s-au ntlnit variaii relativ largi
ale nitriilor reziduali i depiri ale LMA (tabelul nr. 4 i figura nr. 4).
Tabelul 4
Valori ale reziduurilor de nitrii n unele sortimente de salamuri de durat
i procentul depirii LMA

Frecvena
Nr. mg NaNO2/kg
Sortimentul depirii LMA
probe
Intervale Media Abs. Rel.
S. Bnean 15 38,00-62,00 49,33 0 0
S. Choriso 15 32,00-81,00 53,93 1 6,66
S. Sibiu 20 48,00-74,00 64,95 2 10,00
Total/Medie 50 32,00-81,00 56,96 3 6,00

15

13

11 10

9
6,66
% 7 6

1 0

-1
S. Bnean S. Choriso S. Sibiu Media

Figura 4. - Procentul depirii LMA de nitrii n unele


sortimente de salamuri de durat

La aceste produse nitriii reziduali au prezentat limitele cuprinse ntre 32


ppm i 81 ppm NaNO2, iar depiri ale LMA s-au stabilit n procent de: 6,66% la
probele de salam Choriso; 10% la probele de salam de Sibiu i 6% pentru toate
probele cercetate.
La cele 354 probe de produse din carne ca afumturi constituite din 16
sortimente s-au ntlnit depiri ale LMA de nitrii la 11 sortimente (68,75%).
Depiri ale LMA la toate probele s-a stabilit la 4,24% din probe (tabelul nr. 5 i
figura nr. 5).

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Tabelul 5
Valori ale reziduurilor de nitrii n unele sortimente de afumturi
n perioada 2001-2004 i procentul depirii LMA

Frecvena
Nr. Nr. mg NaNO2/kg depirii
Sortimentul
crt. probe LMA
Limite Media Abs. Rel.
1 Bacon 5 28,00-84,00 46,40 1 20,00
2 Costi afumat 29 23,00-108,00 45,90 1 3,45
3 Kaiser 10 10,00-50,10 29,02 0 0
4 Muchi file 40 28,00-79,00 41,66 1 2,50
5 Muchi ignesc 15 29,00-79,00 49,27 1 6,66
6 Muchiule umplut 10 39,00-60,00 48,50 0 0
7 Muchiule Montana 30 30,00-80,00 38,87 1 3,33
8 Pastram Kosarom 10 23,00-28,00 25,60 0 0
9 Pastram porc 55 29,60-88,00 47,36 3 5,45
10 Pastram pui 20 5,00-79,00 39,45 1 5,00
11 Pastram vit 50 36,00-80,00 47,27 2 4,00
12 Rasoale porc 10 24,00-80,00 50,10 1 10,00
13 Rulad extra 10 20,00-40,00 33,30 0 0
14 unc Kosarom 10 21,00-34,00 26,90 0 0
15 unc porc 40 28,00-98,00 48,90 2 5,00
16 unc pui 10 5,00-79,00 36,30 1 10,00
Total/Medie 354 5,00-108,00 43,35 15 4,24

2,5
3,33
3,45
4
5
5
5,45

6,66

10

10
20

0 5 10 15 20
Bacon unc pui Rulad M. ignesc
Pastram porc mg
unc porc NaNO2/kg
Pastram pui Pastram vit
Costi afumat M. Montana Muchi file

Figura 5. - Procentul depirii LMA de nitrii n unele afumturi

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CONCLUZII

Investigaiile ne-au permis s formulm urmtoarele concluzii:


1. La toate sortimentele de produse din carne s-au observat variaii n limite
foarte largi ale nivelului de nitrii reziduali (4 ppm NaNO2 la salamurile afumate
i 121 ppm NaNO2 la prospturi).
2. Depiri ale LMA de nitrii s-a stabilit n procent de 29,38% la
prospturi; 8,99% la crnai; 6,00% la salamurile de durat; 4,24% la afumturi i
3,09% la salamurile semiafumate.
3. Procentul depirii LMA de nitrii la toate produsele s-a cifrat la 9,23%
din produse, mrete riscul asumat prin LMA pentru consumatori.

BIBLIOGRAFIE

1. Banu C., Alexa P., Vizireanu Camelia, 1998 Procesarea industrial a crnii. Ed.
Tehnic, Bucureti.
2. Casens R.G., 1995 Use of sodium nitrite in cured meats today. Food Technol. 49,
72-75.
3. Casens R.G., 1997 Residual nitrite in cured meats. Food Technol. 51, 53-55.
4. Dykhuisen R.S., Frazer R., Duncan C., Smith C.C., Golden M., Benjamin N.,
Leifert C., 1996 Antimicrobial effect of acidified nitrite on gut pathogens:
Importance of dietary nitrite in host defense. Antimicrob. Agents Chemother., 40,
1422-1425.
5. Haldan J., 1901 The red color of salted meat. Journal Hyg. Camb., 1, 115-118.
6. Hansson L.E., Nyren O., Bergstrom R., Wolk A., Lindgren A., Baron J., 1994
Nutrients and cancer gastric risk. A population based caze control study in
Sweden. Int. Journal Cancer, 57, 638-644.
7. Hura Carmen, 1992 Coninutului de nitrai i nitrii din meniurile n Moldova. Rev.
Med. Chirurgical. Iai, 96, 1-2, 41-43.
8. Hura Carmen, 2001 Contaminani chimici n produsele alimentare 1980-2000. Ed.
Cermi, Iai.
9. Ian Elena, 2001 Observaii privind calitatea i salubritatea preparatelor din carne
comercializate pe teritoriul jud. Iai. Lucr. t. USAMV Iai, 44, Fasc. 2, 714-721.
10. Michalski M.M., 1996 Rezidues of nitrites and nitrates in selected meat products
examined in veterinary laboratories within the framework of sanitary-hygiene
supervision in the year 1995. Bull. of the veterinary Institute in Pulawy, Poland,
40, 2, 117-120.
11. Perciballi M., Conboy J.J., Hotchkiss J.H., 1989 Nitrite cured meats as a source
of endogenously and exogenously formed N-nitrosoproline in the ferret. Food
Cosmet. Toxicol., 27, 111-116.
12. Popa G., Segal B., Dumitrache S., Segal RODICA, Apostol C., Teodoru V., 1986
Toxicologia produselor alimentare. Ed. Academiei Romne, Bucureti.
13. Sarasua S., Savitz A.D., 1994 Cured and broiled meat consumption in relation to
childhood cancer. Cancer Causes Control, 5, 141-148.
14. Schuddeboom L.J., 1993 Nitrates and nitrites in Foodstuffs. Council of Europe
Press, Belgium.
15. indilar E.V., indilar E., 2004 Cercetri privind nivelul de nitrai i nitrii n unele
produse din carne. Lucrri t. USAMV Iai, 47, 6, 319-323.
16. indilar E.V., 2005 Contribuii la studiul reziduurilor de nitrai, nitrii i nitrosamine
n unele produse de origine animal. Tez de doctorat, F.M.V. Iai.

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NOI MOLECULE UTILIZATE N TERAPIA ANTIFUNGIC


NEW MOLECULES DESIGNED FOR ANTIFUNGAL THERAPY

M. MARE, Luminia MALIC,


Ingrid APETREI, P. CAZACU, L. MIRON
U.S.A.M.V. Iai

The fungal infections management implies a quick therapy started


very soon after the detection of fungal aetiology. In opposition with
antibiotics designed for bacterial infections, the arsenal of antifungals used
for systemic mycoses is very poor only flucytozine, amphotericin B and
few azoles like fluconazole, itraconazole and voriconazole complied the
necessities of this kind of infections. The use of other new antifungal
compounds like caspofungin micafungin, anidulafungin, posaconazole,
ravuconazole is actually in a period of clinical trials. Antifungal resistance
represents an actua and acute problem because of new trends of significant
increase in strains isolated from superficial and systemic mycoses. In this
paper we discuss four of the most important new antifungals recently
marketed worldwide. The high rate of fungal resistance to current
antifungal compounds has stimulated the research in order to obtain of new
efficient molecules: posaconazole, ravuconazole, caspofungin,
anidulafungin, albaconazole, propiconazole derivatives, mycafungin, FX
0685.

Cuvinte-cheie: antifungice, voriconazol, posaconazol, caspofungin, micafungin.

Micologia medicala este o stiinta relativ noua, a carei dezvoltare in ultimele


decenii este strans legata de cresterea receptivitatii gazdelor la infectia fungica.
Interesul crescand pentru fungii cu potential patogen este relevat si de numarul de
articole publicate in lume pe aceasta tema; astfel, daca in anul 1980 pe PubMed se
puteau inventaria doar 627 de articole despre Candida, in 2001, numarul
articolelor citate de aceeasi retea informationala ajunsese la 2533, adica
aproximativ un articol publicat la fiecare 3 ore. In plus, proportia articolelor
stiintifice care semnaleaza diminuarea sensibilitatii la antifungice a izolatelor
clinice este in continua crestere.
Problematica terapiei antifungice este dominat n zilele noastre de dou
aspecte majore: numrul redus de antifungice disponibile comercial (cca. 25
substane active), pe de o parte, i creterea rezistenei la aceste molecule datorit
remarcabilei plasticiti adaptative a fungilor, pe de alta parte. De remarcat faptul
ca unele micoze emergente (zygomicozele) nu pot fi controlate eficient prin
terapie antifungic, existnd doar o singur molecul activ fa de acestea-
posaconazolul. Costul ridicat al tratamentului cu antifungicele de ultim generaie
destinate omului i animalelor, spectrul selectiv de aciune al acestora, precum i

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toxicitatea rezidual uneori considerabil a celor clasice, reprezint argumente


irefutabile care impulsioneaz cercetarea privind obinerea unor noi substane
active. Un argument n plus la acest demers, l constituie i lipsa oricror mijloace
de profilaxie specific (vaccinare). Dac la acestea adugam variabilitatea extrem
de mare a surselor de infecie (endogene si exogene) i reactivitatea diferit a
indivizilor, care conduc la limitarea drastic a posibilitilor de profilaxie
nespecific, vom constata c, de fapt, arsenalul de lupt mpotriva micozelor este
extrem de srac. n plus, este destul de dificil s luptm cu un microorganism
eucariot (fungul) existent n interiorul unui macroorganism eucariot (omul si
animalele) fr a-i aduce i acestuia prejudicii n urma interaciunii cu aceste
molecule xenobiotice antifungicele. Managementul infeciilor fungice
presupune interventia terapeutic prompt i susinut iniiat ct mai precoce
dup precizarea etiologiei fungice a acestora. Spre deosebire de agenii
antibacterieni (antibiotice), arsenalul terapeutic antifungic utilizabil n infeciile
sistemice este extrem de redus. Alturi de amfotericina B si 5-flucitozina, doar
civa compui azolici ntrunesc rigorile impuse de terapia infeciilor fungice
sistemice: fluconazolul, itraconazolul i voriconazolul. Utilizarea celorlali
compui antifungici de ultim generaie (caspofungina, posaconazol, ravuconazol)
se afl nc n faza de studiu clinic. Rezistena la antifungice reprezint o
problema actual, din ce n ce mai acut, n special pentru clinicieni, datorit
noilor tendine de cretere semnificativ a frecvenei tulpinilor fungice rezistente,
responsabile att de infecii sistemice ct i localizate. Pe lng rezistena genetic
fa de unii agenti antifungici (Candida lusitaniae la amfotericina B, Candida
krusei la fluconazol), actualmente se constat o rezisten dobndit fa de
agentii antifungici n general i fa de unii derivai azolici n special. Aceast
rezisten este mai important n cazul speciilor non-albicans ale genului Candida
(C. glabrata, C.krusei, C. tropicalis, C.parapsilosis, etc.) i aproape
nesemnificativ n cazul tulpinilor de Candida albicans - 2,7% . ntr-un studiu
recent efectuat in Statele Unite pe 2000 tulpini izolate din fungemii, este
semnalat o rat relativ ridicat de rezisten la derivai azolici a izolatelor
aparinnd speciilor non-albicans ale genului Candida comparativ cu cele de
Candida albicans. Astfel, 8% dintre tulpinile de C.glabrata, 8% dintre cele de
C.tropicalis si 34% dintre cele de C.krusei prezentau rezistena la fluconazol, n
timp ce rata rezistenei la acelai antifungic a tulpinilor de C.albicans era de 5%.
Fenomenul se manifest la fel de pregnant i n cazul itraconazolului, fa de care
51% dintre tulpinile de C.glabrata, 12% dintre cele de C.tropicalis i 20% dintre
cele de C.krusei erau de asemenea rezistente, spre deosebire de cele aparinnd
speciei C.albicans la care rata rezistenei era de 8%. n Europa, studii
multicentrice ale susceptibilitii la fluconazol a tulpinilor de Candida albicans
implicate n apariia fungemiilor indic o rata mai redus a rezistenei (2,0-2,7%)
comparativ cu situaia existent in SUA. Fenomenul de rezisten primar i
secundar nu este doar apanajul fungilor levuriformi, el ntlnindu-se i la fungii

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filamentoi de interes clinic (Aspergillus, Zygomicete, etc.). Astfel, s-au semnalat


tulpini aparinnd speciilor genului Aspergillus cu rezisten la itraconazol,
amfotericina B i ali agenti clasici, proporia acestora variind ntre 2-5%.
Proporia crescnd a tulpinilor rezistente a impulsionat cercetarea n domeniul
obinerii de noi molecule dotate cu capacitate antifungic: posaconazol,
ravuconazol, caspofungina, anidulafungina, albaconazol, propiconazol,
micafungina, FX 0685, etc.
Voriconazolul
Voriconazolul este un antifungic triazolic sintetic, de generaia a III-a,
destinat terapiei infeciilor sistemice cu levuri i fungi filamentoi. (Fig.1) A fost
sintetizat de firma Pfizer i n prezent este comercializat sub denumirea de Vfend.
Mecanism de aciune
Ca i ali derivai triazolici, voriconazolul
acioneaz prin inhibiia lanosterol 14- -
demetilazei, blocnd astfel sinteza ergosterolului
necesar edificrii structurilor membranare ale
fungilor.
Spectrul de aciune
Voriconazolul este un antifungic cu
spectru larg de aciune, ce include att fungi
filamentoi, ct i levuri de importan medical.
Trial-urile clinice au indicat eficacitate
terapeutic fa de infecii cu Aspergillus spp.
(A.fumigatus, A.flavus, A.terreus, A.niger,
Fig. 1. Structura chimic a A.nidulans), Scedosporium spp. (S.apiospermum
moleculei de voriconazol i S.prolificans), Fusarium spp. (F.solani,
F.proliferatum .a.) i Candida spp. (C.albicans i alte specii cu rezisten
crescut la fluconazol precum C.glabrata, C.dubliniensis, C.krusei, C.
parapsilosis, C.inconspicua, C.tropicalis .a.). n cazul altor specii (Alternaria
spp., Blastomyces dermatitidis, Blastoschizomyces capitatus, Cladosporium spp.,
Coccidioides immitis, Conidiobolus coronatus, Cryptococcus neoformans,
Exserohilum spp., Exophiala spinifera, Fonsecaea pedrosoi, Madurella
mycetomatis, Paecilomyces lilacinus, Penicillium marneffei, Phialophora spp.,
Scopulariopsis brevicaulis, Trichosporon spp.) s-au obinut rezultate
ncurajatoare, ns datorit numrului redus de cazuri, indicaia terapeutic cu
voriconazol nu este nc bine documentat.
Fa de zygomicete (genurile Absidia, Mucor, Rhizopus etc.), voriconazolul
nu manifest nici un efect terapeutic; n plus, s-a constatat c terapia cu
voriconazol pentru aspergiloza pulmonar invaziv la pacienii neutropenici crete
semnificativ riscul de apariie a zygomicozelor rino-cerebrale.

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Absorbie
Dup administrare oral, voriconazolul se absoarbe rapid i n proporie
ridicat, variaiile pH-ului gastric neinfluennd absorbia sa. Concentraia seric
maxim este atins n 1-2 ore, iar biodisponibilitatea absolut n urma
administrrii orale este estimat la 96%.
Metabolizare
Voriconazolul este metabolizat la nivel hepatic, de ctre izoenzimele
CYP2C19, CYP2C9 i CYP3A4 ale citocromului P450, dar farmacocinetica sa
prezint o variabilitate mare la subiecii umani, din cauza diferenelor genetice.
Toxicitate
n urma efecturii unor studii de toxicitate pe animale de laborator, s-a
demonstrat capacitatea teratogen i embriotoxic a voriconazolului, prin
expunere la doze similare celor terapeutice recomandate la om. Hepatotoxicitatea
se manifest prin creterea nivelului transaminazelor serice, ns ea mbrac doar
arareori un caracter sever. n caz de supradozaj, excesul plasmatic de voriconazol
se poate elimina prin dializ.
Interaciuni medicamentoase
Aceste interaciuni medicamentoase se refer la creterea, respectiv
diminuarea concentraiei plasmatice a voriconazolului ca urmare a influenei
diverselor molecule farmaceutice asupra izoenzimelor citocromului P450.
Compuii cu rol inhibitor pentru aceste enzime (cimetidina, omeprazolul)
determin creterea concentraiei plasmatice maxime a voriconazolului, n timp ce
inductorii enzimatici (rifampicina, carbamazepina, fenobarbitalul, fenitoina,
rifabutina) o diminueaz cu pn la 40-50%.

Posaconazolul
Posaconazolul este un nou derivat triazolic de sintez, cu spectru larg de
aciune, destinat terapiei i preveniei infeciilor fungice invazive (fig. 2). Este
comercializat sub denumirea de Noxafil de ctre firma Shering Plough Inc.
Mecanism de aciune
Inhibiia multiplicrii fungice se explic n cazul posaconazolului, ca i n
cazul altor derivai azolici, prin blocarea unei enzime implicate n sinteza
ergosterolului - lanosterol 14--demetilaza.
Spectru de aciune
Posaconazolul are un spectru larg de aciune care include majoritatea
speciilor genului Candida, Cryptococcus neoformans, Aspergillus spp., Fusarium
spp., zygomicete, feohifomicete i fungi endemici.
n ceea ce privete rezistena la acest nou antifungic, s-au semnalat cazuri
de rezisten ncruciat cu ali derivai azolici (fluconazol, itraconazol) n cazul
unor tulpini de C. albicans (nu ns i n cazul celor de C. krusei) i Aspergillus
fumigatus. Un studiu multicentric desfurat pe parcursul a 10 ani, privind profilul
de sensibilitate la antifungicele de interes major a peste 18000 izolate clinice

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att levuri, ct i fungi filamentoi a relevat efectul salutar al posaconazolului n


cazul majoritii tulpinilor din genurile Candida i Aspergillus care prezentau
rezisten microbiologic la voriconazol, fluconazol i amfotericin B.

Fig. 2. Structura chimic a moleculei de posaconazol

Farmacologie
Posaconazolul este disponibil comercial sub form de preparate
administrabile oral. Absorbia sa este mult mbuntit de co-administrarea
alimentelor i suplimentelor nutritive (de cca. 2-6 ori). Concentraiile plasmatice
terapeutice (cca. 1 g/ml) sunt de obicei atinse n urma administrrii n doze
zilnice 6 mg/kg corp.
Metabolizarea sa are loc preponderent n ficat unde sufer procese de
glucurono-conjugare; metaboliii rezultai sunt inactivi i se elimin n proporie
de peste 70% prin fecale. Excreia renal sczut permite utilizarea
posaconazolului i n cazul pacienilor suferind de diverse nefropatii, la care
administrarea amfotericinei B este contraindicat datorit nefrotoxicitii acesteia.
Dei difuzeaz destul de greu prin bariera hemato-encefalic, studiile
clinice au demonstrat eficacitatea posaconazolului n terapia micozelor sistemului
nervos central.
n cazul acestui antifungic, plasma nu este epurabil prin hemodializ.
Interaciuni medicamentoase
Au fost semnalate interaciuni cu molecule medicamentoase al cror rol de
inhibare a CYP3A4 este binecunoscut. Astfel, clearance-ul posaconazolului se
dubleaz n cazul co-administrrii rifabutinului un inductor al CYP3A4. De
asemenea, administrarea concomitent cu fenitoina sau cimetidina va fi evitat
datorit diminurii concentraiei plasmatice a antifungicului cu 50%, respectiv
40%. n alt sens, sunt necesare precauii cnd posaconazolul este administrat
concomitent cu ritonavirul, indinavirul (le crete concentraia plasmatic).

Caspofungina
Caspofungina este o substan cu rol antifungic obinut prin semi-sintez
n urma modificrii structurale a pneumocandinei B0 o lipopeptid elaborat de

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Glarea lozoyensis (fig. 3). Este comercializat sub denumirea de Cancidas


(caspofungin acetat) de ctre Merck Sharp & Dohme Ltd.
Mecanism de aciune
Caspofungina i exercit efectul antifungic prin inhibarea sintezei de -
(1,3)-D-glucan, o polizaharid-cheie din compoziia peretelui fungilor.
Spectru de aciune
Numrul speciilor inhibate este relativ restrns. Se recomand n terapia
candidozelor i aspergilozelor invazive la aduli. C. parapsilosis i C. krusei sunt
mai puin sensibile la aciunea caspofunginei, iar Cryptococcus neoformans este
rezistent. Este activ i fa de Pneumocystis carinii, fiind singurul antifungic
capabil s-l inhibe.

Fig. 3. - Structura chimic a molecului de caspofungin

Farmacocinetic
Dup administrarea parenteral a unei doze uzuale (70 mg), concentraiile
plasmatice terapeutice se ating n cca. 1 or. Aproximativ 10% din doz persist
n snge timp de 36-48 ore, iar restul se distribuie n diferite proporii n organe i
esuturi. Concentraia cea mai mare se regsete n ficat, la polul opus situndu-se
lichidul cefalo-rahidian n care nu se detecteaz.
Metabolizarea are loc la nivel hepatic, iar metaboliii inactivi rezultai sunt
excretai prin bil (35%) i urin (40%). Doar 1% din doz se elimin
netransformat prin urin. Caspofungina nu poate fi ndeprtat prin hemodializ,
de aceea se impun precauii privind supradozarea la pacienii cu hepatopatii.

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Interaciuni medicamentoase
Doar ciclosporina interfereaz farmacocinetica caspofunginei determinnd
creterea nivelului seric al acesteia, de aceea asocierea celor dou substane nu se
recomand dect n cazuri n care beneficiile obinute exced riscurile.

Micafungina
Micafungina este un antifungic din categoria echinocandinelor, derivat al
unui produs natural izolat din mediul de cultivare al fungului Coleophoma
empetri, recent lansat pe pia sub denumirea de Mycamine (micafungin sodic)
de ctre Astellas Pharma Inc. (fig. 4).

Fig. 4. - Structura chimic a moleculei de micafungin

Mecanism de aciune
Micafungina blocheaz sinteza peretelui celular al fungilor prin inhibiia
sintezei -(1,3)-D-glucanului; acest mecanism particular de aciune permite
inclusiv inactivarea tulpinilor fungice aparinnd genurilor Candida i
Aspergillus, care s-au dovedit rezistente la derivai azolici.
Spectru de aciune
Studiile clinice au confirmat efectul terapeutic al micafunginei n infecii
produse de levuri aparinnd genului Candida (C.albicans, C.glabrata,
C.parapsilosis, C.tropicalis, C.krusei, C.pelliculosa, C.inconspicua, C.lusitaniae)
i fungi filamentoi din genul Aspergillus (cu excepia celor produse de A.
terreus). Efectul profilactic al micafunginei a fost demonstrat n cazul infeciilor
experimentale cu Pneumocystis jiroveci la oareci SCID.

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Farmacocinetica
Concentraiile plasmatice sunt direct proporionale cu doza administrat, iar
timpul mediu de njumtire este de cca. 13,6 ore. Micafungina se distribuie cu
predilecie n pulmoni, ficat, splin i rinichi. Nu este prezent n lichidul cefalo-
rahidian, dei realizeaz concentraii detectabile n esutul nervos.
Proprietile farmacocinetice ale micafunginei sunt similare la pacienii cu
i fr afectarea funciei renale, neexistnd diferene semnificative ale clearance-
ului acesteia. Incidena hepatotoxicitii dup administrarea ndelungat (6 luni)
este mult mai redus (16,6%) n cazul micafunginei dect al amfotericinei B
(100%).
Micafungina este metabolizat de ctre arilsulfataz i catechol-O-
metiltransferaz, iar excreia sa este predominant digestiv (peste 70%). Datorit
gradului nalt de cuplare cu proteinele serice, micafungina nu este o molecul
dializabil.
Interaciuni medicamentoase
Administrarea concomitent a micafunginei cu sirolimus i nifedipin
conduce la creterea concentraiei plasmatice a acestora cu peste 15-18%. De
aceea, pacienii crora li se administreaz concomitent sirolimus sau nifedipin i
micafungin vor fi atent monitorizai, iar dozele celor dou produse
medicamentoase se vor ajusta corespunztor.

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6. Greenberg R.N., Mullane K., Burik J.A., Raad I., Abzug M.J. et al., 2006
Posaconasole as salvage therapy for zygomycosis; Antimicrob. Agents
Chemother; 50(1):126:133
7. Valica V., 2003 - Studiu privind elaborarea unor produse farmaceutice
antiinfecioase. Tehnologie, analiz i standardizare; Tez de Doctor habilitat n
tiine Farmaceutice; Institutul Naional de Farmacie; Chiinu; 2003
8. Miller J.L., Schell W.A., Wills E.A., Toffaletti D.L., Boyce M.et al., 2004 - In vitro
and in vivo efficacies of the new triazole albaconazole against Cryptococcus
neoformans; Antimicrob. Agents Chemother; 48(2):384-387
9. Playfort E.G., Webster Angela, Sorell T.C., Craig C., 2006 - Antifungal agents for
preventing fungal infection in non- neutropenic critically ill surgical patients:

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systematic review and meta-analysis of randomized clinical trials; J. Antimicrob.


Chemother. 57:628-638
10. Morera-Lopez Yolanda, Torres- Rodriguez J.M., Jimenez- Cabello Teresa, Baro-
Tomas Teresa, 2005 - Cryptococcus gattii: in vitro susceptibility to the new
antifungal albaconazole versus fluconazole and voriconazole; Medical Mycology;
43(6): 505-510
11. Takahata S., Okutomi T., Ohtsuka K., Hoshiko S., Uchida K., Yamaguchi H.,
2005 - In vitro and in vivo antifungal activities of FX 0685, a novel triazole
antifungal agent with potent activity against fluconazole- resistant Candida
albicans; medical Mycology; 43(3): 227-234
12. Gonzalez G.M., Fothergill W.A., Sutton D.A., Rinaldi M.G., Loebenberg D., 2005 -
In vitro activities of a new and established triazoles against opportunistic
filamentous and dimorphic fungi; Medical Mycology; 43(3): 281-284
13. Torres H.A., Hachem R.Y., Chemaly R.F., Kontoyiannis D.P., Raad I.I., 2005 -
Posaconazole: a broad spectrum triazole antifungal; Lancet Infect. DIS., 5:775-
785
14. Jarvis B., Figgitt D.P., Lesley J.S., 2005 - Micofungin; Drugs; 64(9): 969-982
15. Di Domenico B., 1999 - Novel antifungal drugs; Current opinion in Microbiology,
2:509-515

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GRADUL DE CONTAMINARE CU MICETI I


MICOTOXINE A UNOR SUBSTRATURI VEGETATALE
FOLOSITE N HRANA OMULUI I A ANIMALELOR
I. COMAN, Rodica CUCIUREANU,
Delia BULEA, Maria BOBUAC,
Alina TEODOR, Luminita MALIC
U.S.A.M.V. Iai

Au fost analizate micologic si micotoxinic 82 de probe de substraturi


vegetale reprezentate de: arahide neprajite vrac(9), cafea verde vrac(6),
porumb boabe (38), soia boabe (8), seminte de dovleac (4), seminte de
floarea soarelui (6), arahide prajite in cutii vidate (8) si furaje combinate
(3).
Investigatiile intreprinse au evidentiat o contaminare fungica extrem
de variata,numarul unitatilor formatoare de colonii osciland intre 0-
1400g/produs ij functie de natura sortimentului,de conditiile de stocare si
tehnologiile de procesare.Astfel,incarcatura micotica a inregistrat valori
cuprinse intre 910 si 280 10 in probele de arahide neprajite,intre
1510 si 11010 in esantioanele de cafea verde,intre 0 si 13410 in
boabele de soia,intre 4 10-140010 in probele de porumb ,intre 1110-
14910 in seminte de dovleac si intre 510 si 28010 respectiv 9710
si 46910 in seminte de floarea soarelui si nutret combinat.
Examenul micotoxinic efectuat pe toate cele 82 de probe a permis
izolarea,separarea si identificarea unui numar de 11 micotoxine dintre care
2 au fost confirmate ca aflatoxina B1 si aflatoxina G1,in una din probele de
furaj combinat,5 dintre acestea s-au dovedit a fi precursorul acestora,si
anume sterigmatocistina in boabe de cafea verde(3) si boabe de porumb(2)
iar 4 spoturi fluorescente au fost incadrate pe baza fluorescentei si al Rf-
ului in clasa achratoxinelor;acestea au fost prezente in probele de porumb
boabe(2) si boabe de soia (2)

Amploarea fenomenului de contaminare micotoxinic este att mare nct


se poate afirma fr teama de a fi greit c, micotoxinele au devenit un poluant
planetar de vreme ce ele au fost evideniate pe toate meridianele lumii, din
inuturile reci i umede ale Rusiei (Siberia) pn n zonele aride ale Africii de
Sud, din Regatul Unit al Marii Britanii pn n arhipelagul Japoniei, n Australia
i Noua Zeeland.
Aceste afirmaii, ndrznee poate sunt susinute de cercetri riguroase care
au evideniat micotoxine n boabele de cereale (gru, orz, porumb, ovz, org,
secar) i leguminoase (soia, arahide, mazre, fasole), n toate produsele i
subprodusele acestora (pine, aluat, prjituri, franzelue de mlai), n seminele de
oleaginoase ( floarea soarelui, bostan, miez de nuc), n legume i fructe (morcov,
ardei, tomate, ptrunjel, mere, pere, gutui, citrice, smochine, kiwi, piersici, caise),

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n preparatele acestora (gem, dulcea, compot, peltea, sucuri naturale), n bere,


must, vin, cidru, n condimente (piper verde, negru, alb), paprica, currey, oregano,
cimbru, boia de ardei i n plantele medicinale (frunze de ptlagin, suntoare).
Opinia public din rile civilizate a fost informat i chiar cu amanunte
mai puin semnificative,asupra pericolului pe care agentii conservani, aditivii,
insecticidele, fertilizanii chimici, pesticidele n general, l reprezint pentru
sntatea animalelor i implicit a omului. Din pcate se trece sub tcere n mod
inexplicabil posibilitatea contaminrii alimentelor i a furajelor cu micromicei
toxigeni i riscurile pe care infestaiile respective le incumb.
La ora actual micotoxinele nu mai pot fi considerate doar factori poteniali
de risc pentru sntatea omului intrucat ele reprezint adevraii ageni cauzatori
ai unor entiti morbide grave, deseori cu sfrit letal,considerate deja ca boli cu
statut propriu precum aflatoxicoza, ochratoxicoza, nefrita endemic micotoxinic,
sindromul Reyes, carcinomul hepatocelular primar, Aleukia toxic alimentar,
boala Kershin-Berk, cancerul esofagian fumonisinic, Akakabi-byo,etc.

MATERIALE I METODE DE LUCRU

Au fost analizate prin examen micologic cantitativ i calitativ precum i prin


investigaii micotoxinice 82 probe recoltate din reeaua de comercializare sau din
ferme de animale, reprezentnd substraturi vegetale folosite n hrana omului i a
animalelor.
Pentru izolarea i evaluarea numeric a micromicetelor s-au folosit tehnici
uzuale de laborator i medii de cultivare adecvate (PDA, Sabouraud), iar pentru
identificarea i ncadrarea taxonomic a speciilor dominante de micromicei s-au
coroborat aspectele culturale ale coloniilor cu particularitile morfostructurale ale
corpilor fructificani. Izolarea, separarea i identificarea micotoxinelor fluorescente
s-au realizat prin tehnica cromatografiei n strat subire (CSS).

REZULTATE I DISCUII

Au fost examinate micologic i micotoxinic 82 probe de substraturi


vegetale reprezentnd arahide neprjite vrac (9), cafea verde vrac (6), porumb
boabe (38), soia boabe (7), semine de dovleac (4), semine de floarea soarelui (6),
arahide prjite n cutii vidate (8), furaj combinat (3).

Examenul micologic cantitativ


Investigaiile ntreprinse au evideniat o contaminare fungic absolut
inegal, numrul unitilor formatoare de colonii (UFC) oscilnd ntre zero i
1400103/g produs, n funcie de natura sortimentului, de condiiile de stocare i
tehnologiile de procesare (tabelul 1)

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Tabelul 1
Rezultatele examenului micologic cantitativ si calitativ

Proportia genurilor de micei identificate


Nr. Limitele de
Nr. n probele analizate
Sortiment probe variatie ale
crt Alte
analizate UFC(10/g) P A F Cl Lv
genuri
Arahide
1 neprajite 9 9-280 33,3 66,6 11,1 11,1 100,0 55,5
vrac
Cafea verde
2 6 15-110 100,0 66,6 0 66,6 0 83,3
vrac
Porumb
3 38 4-1400 76,3 60,5 73,6 26,3 17,9 63,1
boabe
4 Soia boabe 7 0-134 57,1 71,4 14,8 28,5 42,8 71,4
Seminte
5 4 11-149 50,0 25,0 75,0 50,0 50,0 100,0
dovleac
Seminte
6 floarea 6 5-280 66,6 66,6 50,0 50,0 16,6 50,0
soarelui
Arahide
7 prajite in 8 0-50 0 100,0 0 0 0 0
cutii vidate
Furaj
8 3 97-469 66,6 66,6 66,6 33,3 33,3 33,3
combinat
TOTAL
9 82 0-1400
PROBE

Legenda:
P Penicillium; A Aspergillus; F Fusarium; Cl Cladosporium; Lv - Levuri

ncrctura micotic a nregistrat valori cuprinse ntre 9103 i 280103/g


n probele de arahide vrac neprjite, ntre 15103/g i 110103/g n probele de
cafea verde vrac, ntre zero i 134103/g n probele de soia , ntre 4103/g i
1400103/g, respectiv 11103 /g i 149103 /g n probele de porumb si seminele
de dovleac, ntre 5103/g i 280103/g n cele de floarea soarelui, ntre zero i
50103/g n probele de arahide prjite i comercializate n cutii ermetizate i ntre
97103 /gi 469103/g n nutreurile combinate. Cele mai poluate eantioane din
punct de vedere micologic au fost cele de porumb boabe, dar se cuvine s
accentum n acelai timp faptul c, arahidele prjite i comercializate n cutii
vidate au fost lipsite de fungi levuriformi i filamentoi, cu excepia unei singure
probe care la examenul organoleptic s-a dovedit de asemenea necorespunztoare,
cutia fiind bombat. Cu siguran ca aceasta a prezentat o fisur la nivelul creia
s-a i produs contaminarea.

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Examenul micologic calitativ


Prin protocolul iniial de lucru s-a prevzut s fie identificate i evaluate
catitativ genurile de micromicei implicate mai frecvent n patologia omului i a
animalelor. Aceast conduit intuitiv i-a dovedit ntr-adevr valabilitatea, ea
fiind confirmat de rezultatele obinute i prezentate n tabelul 1. Singurul
sortiment care a fcut excepie i a etalat o alt palet micotic au fost seminele
de dovleac, a cror contaminare fungic a fost dominat de genuri neidentificate,
care au atins cota procentual maxim 100%. Se impune deci, n cazul acestui
parametru biologic, extinderea investigaiilor micologice, pentru catagrafierea
corect a speciilor de micromicei ce-l caracterizeaz, cu att mai mult cu ct,
seminele de dovleac s-au comportat diferit comparativ cu boabele de soia i
seminele de floarea soarelui la aciunea unor ageni chimici utilizai ca elemente
conservante (Coman i col. date n curs de publicare).
Exprimarea procentual a genurilor dominante de micromicei a evideniat
limite de variaii extrem de largi, n funcie de natura substratului i apartenena
taxonului.
Speciile aparinnd genului Penicillium au fost prezente n toate probele de
cafea verde i au oscilat procentual ntre zero i 76,3% n celelalte substraturi.
Micromiceii ncadrai n genul Aspergillus au fost contaminani solitari ai
arahidelor prjite i pstrate n cutii vidate i au avut valori cuprinse ntre 25,0%
i 71,4% n probele celorlalte sortimente vegetale. Extrem de interesant este
repartiia genului Fusarium n eantioanele analizate, deoarece, n cafea, arahide i
soia micromiceii acestui gen au fost abseni sau numrul lor a fost foarte mic
(11,1%, respectiv 14,8%), n timp ce n boabele de porumb sau n seminele de
dovleac gradul de reprezentabilitate a atins valori superioare de 73,6% sau 75,0%.
Aceast repartiie inegal este justificat foarte probabil de gradientul de
umiditate, psichrofilia fungilor ncadrai n genul Fusarium fiind recunoscut i
dicteaz, iat, intensitatea fenomenului de multiplicare a formelor vegetative, a
aparatului micelian n totalitate.
Cladosporium s-a dovedit nc o dat un contaminant discret dar constant al
substraturilor vegetale i aceasta datorit supleei sale ecologice care i permite s
supravieuiasc n condiii de mediu foarte diferite.
Fungii levuriformi de tipul Candidei i Rodotorulei i-au fcut simit
prezena n unele sortimente, dar au lipsit din cafea i arahidele vidate. Surprinde
ns gradul foarte ridicat de contaminare (100%) al probelor de arahide neprjite,
care au fost meninute probabil n spaii de conservare improprii.

Examenul micotoxinic
Micotoxinele sunt metabolii fungici secundari cu aciune nociv extrem de
sever pentru toate sistemele biologice cunoscute. Ele afecteaz metabolismul
trofinelor, fiind implicate direct n tulburrile funcionale i structurale ale unor

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organe vitale (ficat, rinichi), dar i statusul imunitar ce vizeaz deopotriv


elementele celulare i umorale.
Rezultatele examenului micotoxinic au evideniat n mod surprinztor
prezena n substraturile analizate a unor micotoxine care pot afecta grav sntatea
omului i a animalelor (Tabelul 2). Vom sublinia n primul rnd prezena n cafea
a sterigmatocistinei, care este precursorul aflatoxinei B1, ambele micotoxine fiind
implicate frecvent n stri morbide grave la om i animale. Faptul c 50% dintre
probe au fost contaminate cu aceste micotoxine demonstreaz gradul ridicat de
contaminare micotoxinic al boabelor de cafea verde i necesitatea introducerii
unui program strategic preventiv, prin care se vor impune condiiile optime de
recoltare transport i conservare a boabelor de cafea, controlul calitativ al acestora
,prin examene micologice i micotoxicologice periodice precum i perfectionarea
tehnologiilor de procesare a boabelor verzi de cafea.
Tabelul 2
Rezulatele examenului micotoxinic

Proportia genurilor de miceti


Nr. Nr.probe Nr. Spoturilor
Sortiment identificate inprobele analizate
crt. analizate fluorescente
AF ST O ZEA N.I.
Arahide
1 9 0-2 - - - - -
neprajite vrac
Cafea verde
2 6 2-6 - 3 - - -
vrac
3 Porumb boabe 38 0-5 - 2 2 - -
4 Soia boabe 7 0-3 - - 2 - -
5 Seminte dovleac 4 0-2 - - - - -
Seminte floarea
6 6 0-4 - - - - -
soarelui
Arahide prajite in
7 8 0 - - - - -
cutii vidate
8 Furaj combinat 3 1-5 2 - - - -
9 TOTAL PROBE 82 0-6 2 5 4 - -

Legenda: AF Aflatoxine; ST Sterigmatocistina; O Ochratoxina;


ZEA Zearalenona; N.I. - Neidentificate

Boabele de porumb s-au dovedit sortimentul cel mai poluat micotoxinic,


dou probe fiind contaminate cu strigmatocistin iar altele dou cu ochratoxina A.
Avnd n vedere faptul c aceste substraturi vegetale intr n proporie mare n
compoziia nutreurilor combinate, riscul ca micotoxinele respective s ajung n
hrana animalelor devine iminent. Ipoteza este susinut i de faptul c, una dintre
micotoxine i anume sterigmatocistina se regsete n furajul combinat sub forma
sa final de aflatoxin.
n boabele de soia au fost puse n eviden spoturi fluorescente dintre care
dou au fost identificate ca fiind ochratoxina A.

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CONCLUZII

Au fost examinate micologic i micotoxinic 82 probe de substraturi


vegetale recoltate din reeaua de comercializare i fermele de animale.
1. Examenul micologic cantitativ a evideniat o contaminare fungic
inegal, numrul unitilor formatoare de colonii (UFC) oscilnd ntre 0-
1400103/g n funcie denatura sortimentului i tehnologia de procesare a
acestuia. Cele mai poluate au fost boabele de porumb, iar cel mai puin
contaminate arahidele prjite n cutii vidate.
2. Flora micotic evideniat a fost dominat de fungii filamentoi
aparinnd genurilor Penicillium, Aspergillus, Fusarium, Cladosporum, dar i de
micromiceii levuriformi de tipul Canidei albicans.
3. Investigaiile micotoxinice au permis izolarea, separarea i identificarea
unor micotoxine fluorescente de tipul sterigmatocistinei n boabele de cafea verde
i boabele de porumb, a ochratoxina A n boabele de porumb i soia, i a
aflatoxinelor n probele de furaj combinat.

BIBLIOGRAFIE

1. Akyama, H., Kikuchi Y., Chen, D., Goda, Y., Takatoki, K, Ichinoe, M., Toyoda, M.,
1998 A rapid analytical method of ochratoxin A (OTA) in food and natural
contamination in read pepper. Revue Med. Vet., 149, 6:493
2. Azemar, B., Pinelli, E., Plante, P., Escourron, G., Petkova-Bocharova, T., Pfolil-
Leszkowicz, A., 1998 Some human kidney tumors in France exhibited a
specific ochratoxin A DNA adduct pattern. Revue Med. Vet., 6:653
3. Battilani, P., Pietri, A., 2002 Ochratoxin A in grapes and wine. Eur. J. Plant
Pathol., 108:638
4. Blanc, M., Pittet, A., Munoz-Box, R., Viani, R., 1998 Behavior of ochratoxin A
during green coffee roasting and soluble coffee manufacture. J. Agric. Food.
Chem., 46:673
5. Castegnaro, M., Mc Gregor, D., 1998 Carcinogenic risk assessment of
mycotoxins. Revue Med. Vet., 149,6:671
6. Coman, I., Mare, M., Ancua Apreutesei, 2002 Rolul i importana epidemico-
sanitar a micotoxinelor. Lucr. t., fascicula II, Ed. Ion Ionescu de la Brad, Iai,
Vol. 45 (4):457
7. Coman, I., Popescu, O., 1985 Micotoxine i micotoxicoze. Ed. Ceres, Bucureti.
8. Council for Agricultural Science and Technology cost, 1989: Mycotoxins:
economic and health risks. Task Force Report, Anes, Iowa, 91
9. Majerus, P., Bresch, H., Ottender, H., 2002 Ochratoxin A in wines, fruit juices and
seasonings. Achiv. Fur Lebensmittelhygene, 51:95
10. Pittet, A., 1998 Natural occurrence of mycotoxins in food and feeds an update
review. Revue Med. Vet., 149:479
11. Tanaka, T., Hasegawa, A., Yamamato, S., Lee, U., Sugiura, Y., Ueno, Y., 1988
Worldwide contamination of cereals by the Fusarium mycotoxins nivalenol,
deoxinivalenol and zearalenone. Survey of 19 countries, J. Agric. Food. Chem.,
36:979

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THE PREVENTION OF FUNGI AND MYCOTOXIN


CONTAMINATION OF OILSEEDS BY CHEMICAL
CONSERVATION USING VOLATILES REAGENTS
I. COMAN, Alina SRBU,
Luminia MALIC, Maria BOBUAC
U.S.A.M.V. Iai

INTRODUCTION

Oilseeds, like soybeans, sunflower seeds and pumpkin seeds, are exposed
to the mycotoxins attack, as much as the grains are.
It is enough to remember the dbut of the turkey X disease in England,
which result was the intoxication and the death of more than 100.000 turkeys,
after the ingestion of soy grist contaminated with Aspergillus flavus and aflatoxin
B1.
The local and international researches, concerning the conservation
possibilities of oilseed with high levels of moisture, are few, incomplete and
convinctionless.
Nearby the important factories of mixed feed there are laboratories, which
have pharmaceutical formula with mycostatic activity. This formula inhibits the
mycets development, but its effect is only on the final product until its
consumption. The raw materials used in mixed feed or in silo preparation
represent the true critical mass, which has to be conserved in order to prevent the
mycets development and the mycotoxin production.
This was the aim of this study, to prevent the fungi contamination by
conservation of oilseeds with high levels of humidity, using unpolluant, volatile
chemical substances in order not to affect their nutritive value.

MATERIALS AND METHODS

1. Oilseed samples, micromycets strains, medium


Soybeans, sunflower seeds and pumpkin seeds were taken from the field
or silo spaces and were analyzed immediately in the lab in order to establish the
humidity level and their nutritive value by usual procedures.
The ten strains, most commonly found on grains, were selected from the
collection of the Mycology and Mycotoxicology Laboratory of Veterinary Medicine
Faculty from Iassy. These dominant strains were: Aspergillus flavus, Aspergillus
niger, Aspergillus fumigatus, Alternaria alternata, Fusarium graminearum,
Fusarium culmorum, Stachybotris alternans.
As medium for these cultures development were used Sabouraud and
Potates Dextrose Agar.
2. Experimental methods

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Inocul preparation
To obtain the biological material, the 10 strains of fungi were seeded in test
tubes with inclined medium for 8 days at 25C, then they were passed-by 10
Roux-dishes in 200 ml Sabouraud medium and incubated in darkness at 25C.
Every three days, the cultures were analyzed in progress to observe their
phenotypic particularities and the development of spore, conidia and
chlamydospore.
After 25-28 days, when the sporulation was at its high level, in each Roux
dish were introduced 50 ml physiological serum, which helped the obtaining of
strains.
These were centrifuged at 2500 rot/min. for 30 min. and the sediment was
resuspended into an equivalent quantity of physiological serum. All the contents
were introduced into an Erlenmeyer balloon and then kept at 4C. That
represented the source of the biological material.
Experimental model
33 samples, each of 100 grams, were analyzed in the experiment. Three
samples of each assortment represent the witness: the first one with seeds only,
the second with seeds and 2 ml sterile physiological serum and the third one with
seeds contaminated with 2 ml of biological material.
The next 3 samples were contaminated with 2 ml of selected strains and
with 1 ml pure acetic acid at different concentration: 1%, 0,75% and 0,50%.
Other 3 samples, contaminated with 2 ml inocul were treated with 1 ml
formic acid at the same concentrations.
The last 3 samples, also contaminated with 2 ml of biological material were
treated with pure acetic acid and formic acid aa, the mixture being at 1%, 0,75%
and 0,50% concentrations. The pumpkin seeds were not treated with this last
mixture.
All the 33 samples were introduced in boxboards and kept at room
temperature for 9 months.
The fungi dynamics of each sample was observed every month through
quantitative and qualitative mycological exams.

RESULTS AND DISCUSSION

Table 1 shows the results of mycotical charge of soybeans, sunflower seeds


and pumpkin seeds treated with pure acetic acid, formic acid and their
combination at different concentrations during the 9 months experiment.
The pure acetic acid, which the soybeans were treated with, even at its
weakest concentration (0,50%), had a severe destructive effect upon the spore of
micromycets.
Thus, at 1% acid concentration, the number of CFUs were among 10-760,
at 0,75%, between 10-830 and at 0,50% oscillated among 170-1600.
The percentage reduction of mycotical charge of soybeans treated with
formic acid at 0,50, 0,75% and 1% concentration was extremely severe, having
values among 99,66% and 100%.

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The number of the cells which survived the chemical treatment at 1%


formic acid was between 0-40 and at 0,75% and 0,50% among 0-210.
The cid effect of pure acetic acid and formic acid mixture was powerful,
being in proportion of 98, 85%-100%.
The efficiency of sunflower seed treatment with pure acetic acid was
remarkable and it was between 96,26%-100%, the number of CFUs oscillating
among 0-410 at 1% concentration, 80-4760 at 0,75% acid and 180-2230 at 0,50%.
The formic acid reduced the CFUs in proportion of 99,87%-100% and the
mixture with pure acetic acid demonstrated their compatibility, results revealed
also by the destructive effect of this formula upon the micromycets growth. Thus,
the number of CFUs oscillated among 0-605, depending on mixture
concentration, the percentage being between 99,76% and 100%.
It seems that the formic acid is more effective than pure acetic acid,
because of its destructiveness upon the entire fungal charge, which was observed
in 22,20% sunflower samples treated with formic acid and only in 3,6% samples
treated with pure acetic acid.
The reduction of CFUs from pumpkin seeds treated with pure acetic acid
was only of 49,08%-90,90% at 1% concentration, between 31,36%-91,91% at
0,75% acid and 18,32%-73,08% at 0,50% concentration.
After formic acid treatment, the number of CFUs was between 35,73%-
99,23%, depending on its concentration, subordinate those from soybeans and
sunflower seeds. Also, the seeds coat became grey or dark, sometimes with red
spots. Thus, these seeds could not be protected against fungi and mycotoxin
contamination with these chemical reagents, which were successfully used upon
soybeans and sunflower seeds.
The qualitative mycological investigations revealed that, from all 10 strains
of fungi, every month were observed constantly Aspergillus fumigatus,
Aspergillus niger, Aspergillus flavus, Penicillium spp., alternating with the other
species of Fusarium, Cladosporium, Alternaria, Stachybotris and Mucor-
unselected strains.
The fungal mosaic, in its variety, is shown below in the Petri dishes from
figures 1, 2 and 3.
Witness+ inocul Acetic acid 1% Acetic acid 0.5%

Formic acid 1% Formic acid 0,5% Mixture 1% Mixture0,5%

Fig.1 Inhibitory effects of chemical treatment of soybeans

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Witness+ inocul Acetic acid 1% Acetic acid 0.5%

Formic acid 1% Formic acid 0,5% Mixture 1% Mixture0,5%

Fig. 2 Inhibitory effects of chemical treatment of sunflower seeds

Witness+ inocul Acetic acid 1%

Acetic acid 0.5% Formic acid 1% Formic acid 0,5%

Fig. 3 Inhibitory effects of chemical treatment of pumpkin seeds

CONCLUSIONS

The researches intended to preserve soybeans, sunflower seeds and


pumpkin seeds with high levels of moisture (16-27%) by acid treatment at 1%,
0.75%, 0.50% concentration, in order to destroy the mycotical charge and to
prevent the mycotoxin contamination.
The pure acetic acid and formic acid assured a great conservation in time of
soybeans and sunflower seeds, which remained unalterable as aspect, odor, color
and shape during the entire period of experiment.
The qualitative mycological exams revealed the severe destructive effect of
these two acids single or mixed, which reduced the number of CFUs to 98.77%-
100% on soybeans and to 96.26%-100% on sunflower seeds. These values were
stable during the entire experiment.
The most acetic and formic acid resistant were the Aspergillus fumigatus,
Aspergillus niger, Aspergillus flavus and Penicillium notatum. These species were

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

constantly found in the analyzed samples, but their activity during the 9 months
was not explosive.
The chemical treatment is not efficient on pumpkin seeds, because of its
reduced proportion of destroyed fungi and the alteration of seeds coat, which
became grey or dark with red spots on it.

REFERENCES

1. Ainsworth, G.C., 1971 - Dictionary of fungi, Ken Surrey.


2. Alexopoulos, C.J., 1979 - Introductory mycology, 2end Ed., John Wiley and
sons(ed), New York.
3. Battilni, P., Pietri, A., 2002 - Ochratoxin A in grapes and wine. Eur. J. Plant Pathol.
4. Blanc, P., Pittet, A., Minoz-Box, R., Viani, R., 1998 - Behaviour of ochratoxin A
during green coffee roasting and soluble coffee manufacture. J. Agric. Food
Chem.
5. Bontea, Vera, 1985-1987 - Ciuperci parazii n Romnia, Ed. Acad. R.S. Romania,
Vol.1 and 2, Bucureti.
6. Booth, C., 1971 - The genus Fusarium. Commonwealth Mycological Institute. Kew,
Surrey, England.
7. Coman, I., Popescu, O., 1985 - Micotoxine i micotoxicoze, Ed. Ceres, Bucureti.
8. Coman, I., Mare, M., Apreutese, Ancuta, 2003 - Aspecte inedite privind
patogeneza i exprimarea clinic a intoxicaiei cu fumonisine. Rev. Romn de
Med. Vet.
9. Coman, I., Mare, M., Apreutese, Ancuta, 2002 - Rolul i importana epidemico-
sanitar a micotoxinelor. Lucr. t. Fascicula II, Ed. Ion Ionescu de la Brad, Iai.
10. Coman, I., Popescu, O., Bazgan, Olimpia, 1983 - Micotoxinele, problem actual a
patologiei veterinare. Cercet. Agron. n Moldova.
11. Coman, I., Popescu, O., Grundberg, N., 1978 - Separarea i identificarea
cromatografic a unor micotoxine din materii prime furajere. Rev. De Creterea
anim.
12. Coman, I., Popescu, O., Halga, P., Grundberg, N., 1977 - Evidenierea
cromatografic a unor micotoxine din furaje. Simpozion Micotoxicozele la
animale, Iai.
13. Coman, I., Popescu, O., Halga, P., 1987 - Aprecierea igienic a furajelor prin
investigaii micotoxicologice. Cercet. Agron. n Moldova.
14. Costa, L.L.F., Scussel, V.M., 1998 - Toxigenic fungi in beans. Symposium
International Mycotox, Toulouse.
15. Drochner, M., 1989 - Prezena fusariotoxicozelor n furaje. Dtsch. Tierarztl, Wschr.
16. Gilbert, J., 1989 - Current views on the occurance and significance of Fusarium
toxins. J. Appl. Bacteriol. Symp. Suppl.
17. Holzapfel, C.W., 1968 -The isolation and structure of cyclopiazonic acid and toxin
metabolite of Penicillium cyclopium Westling Tetrahedron.
18. Hulea, Ana, Iliescu, P., 1994 - Determinator pentru identificarea mucegaiurilor
potenial patogene. Societatea de Med. Vet. Din R.S. Romnia, Bucureti.
19. Iacob, Viorica, 1994 - Fitopatologie, curs, Iai.
20. Lillhoy, E.B.R., Garcia, W.J., Lambrow, M., 1974 - A. flavus infection and aflatoxin
production. Influence of trace elements. Appl. Microbiol.
21. Mititiuc, M., 1995 - Micologie. Ed. Univ. Al. I. Cuza, Iai.
22. Yazdanpanah, H., Daraci, B., Esmaeil-Zadeh, F. - Natural occurence of Fusarium
mycotoxins in 21 infants grain/ based food samples from Jiranian market. Revue
Med. Vet

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CERCETRI PRIVIND EVALUAREA BUNSTRII


GINILOR OUTOARE EXPLOATATE N SISTEM
ALTERNATIV, NTR-O FERM DIN ZONA DE SUD
A ROMANIEI
RESEARCHES CONCERNING THE WELFARE ASSESSMENT OF THE FREE-RANGE
LAYING HENS RAISED IN A UNIT FROM SOUTHERN ROMANIA

F. FURNARIS, Elena MITRNESCU,


L. TUDOR, B. TABAC, C. TOGAN

The researches had in view the assessment of animal welfare level in


a farm of laying hens raised in alternative system, in the southern area of
Romania - Olt County. The assessment consist of a single farm visit, that
took place in 2005, the 16th of September, being applied the ANI (Animal
Needs Index) 35L system in order to establish the welfare of 3500 hens
housed in one shelter, with outdoor access.
The main argument of choosing the above mentioned assessment
method among others integrative numerical systems of hens welfare
assessment (e.g. swede system, english system launched in Bristol Welfare
Assurance Program, german system ANI 200) was its rapidity. The method
consists in combining as a unique result engineering-based parameters
(details concerning shelter architecture and endowments) with animal-
based parameters (physiological e.g. feathers condition, skin condition or
ethological e.g. number of cocks in the flock of hens). The parameters,
gathered in 5 area of influence locomotion, social interaction, flooring,
light and air, stockmanship were established either by measurements and
direct observation of the flocks, or by last generation devices as BK 2250
sonometer (used to assess noises level) and Drager Miniwarn gas-meter
(used to assess air quality). In order to assess the parameters feathers
condition and skin condition were used up-to-date methods, which are
based on awarding scores comparing the on-the-field aspects with photo
guides.
Every parameter receives a score which are added so that the final
ANI 35 score can be established. The hens' welfare final score at the farm
studied recorded 23.5 points. This value shows that housing conditions and
flock management are fairly suitable with respect of welfare (final score
between 21 and 24 points), a favorable and beneficent situation.

Problema bunstrii animalelor nu mai reprezint pentru societatea uman


contemporan un deziderat sau o perspectiv, ci, din ce n ce mai acut, o
necesitate stringent.
n acest context, Romnia i-a crescut n ultimul timp exigenele fa de
problema bunstrii animalelor, att din perspectiva aderrii la Uniunea

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European, ct i din necesitatea meninerii n comerul internaional cu animale


vii i cu produse de origine animal.
Cercetri similare celor care fac obiectul acestei lucrri trebuie s se
realizeze la scar larg n ara noastr, pentru a se crea n timp o baz de date cu
suficiente nregistrri, fr de care nu poate fi stabilit pe baze obiective nivelul de
bunstare a animalelor acceptat n mod oficial i nu poate fi creat un sistem
propriu de evaluare a bunstrii animalelor, adaptat necesitilor din teren.
Lucrarea de fa i propune dou obiective: s testeze aplicabilitatea unui
sistem austriac de evaluare a bunstrii animalelor n condiiile Romniei i s
demonstreze nivelul ridicat de bunstare a ginilor outoare exploatate n sistem
alternativ, n condiiile unei productiviti comparabile celei din sistem intensiv.

MATERIALE I METOD

Deoarece Romnia nu dispune nc de un sistem numeric propriu de


evaluare a bunstrii ginilor outoare, am utilizat un sistem de evaluare
oficializat n Austria, respectiv sistemul ANI 35L Animal Needs Index 35L.
Principalul argument care a condus la alegerea acestuia din ntreaga gam
a sistemelor numerice integrative (sistemul suedez, sistemul englez din cadrul
Bristol Welfare Assurance Program, sistemul german ANI 200) a fost reprezentat
de rapiditatea n aplicare. Sistemul ANI 35 const n combinarea sub forma unei
unice rezultante a factorilor de bunstare engineering based (detalii privind
arhitectonica adposturilor i dotrile acestora) cu factorii animal based
(fiziologici: starea penajului, starea pielii sau comportamentali: numrul de cocoi
din efectiv).
Factorii relevani pentru bunstarea psrilor, clasificai conform ANI 35L n
5 grupe de factori, s-au apreciat fie prin observaii directe n efectivele de psri i
msurtori metrice, fie apelnd la aparatur modern de tipul sonometrului BK
2250 (utilizat pentru determinarea factorului zgomotele din adpost) sau a
gazometrului electronic Drager Miniwarn (utilizat pentru determinarea factorului
calitatea aerului). Pentru aprecierea unor factori fiziologici (starea penajului,
starea pielii) s-au folosit metode numerice moderne de apreciere, care au la baz
acordarea de note prin comparare cu ghiduri foto.
S-au acordat note pentru fiecare factor al grilei de evaluare, iar prin
nsumare s-a obinut punctajul ANI final pentru efectivul de psri evaluat. Pe
baza punctajului s-a interpretat nivelul bunstrii psrilor exploatate n sistem
alternativ.

REZULTATE I DISCUII

Punctajul final ANI obinut a fost de 23,5 puncte; ca urmare bunstarea


ginilor outoare exploatate n sistem alternativ se nscrie n categoria bunstrii
satisfctoare.

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Notele acordate pentru fiecare factor din cadrul grilei de evaluare ANI 35L
pentru gini outoare sunt prezentate n tabelele nr. 1-5.
Tabelul 1
Tabelul de evaluare a primei grupe de factori libertatea de micare

Calificativul sau valoarea Notele


Factorii evaluai
msurat/observat acordate
2
Suprafaa minim disponibil 0,18 m /gin 0,5 puncte
Raportul dintre suprafaa de relaxare i 25,03% - 0,5 puncte
scormonit i suprafaa pardoselii
2
Suprafaa de micare n padoc 0,353 m /gin 0,5 puncte
Accesul n padoc n zile/an 255 zile 0,5 puncte
2
Suprafaa de teren nierbat 3,18 m /gin 0 puncte
Distana maxim ntre adpost i limita 130 m 0 puncte
zonei nierbate
PUNCTAJUL GENERAL/GRUP 1 punct
LIMITELE DE VARIAIE ALE PUNCTAJULUI GENERAL/GRUP - 2 9 puncte

Tabelul 2
Tabelul de evaluare a grupei a II-a de factori interaciunile sociale

Calificativul sau valoarea Notele


Factorii evaluai
msurat/observat acordate
Mrimea lotului 1698 gini/arc 0 puncte
Suprafaa minim disponibil 0,18 m2/gin 0,5 puncte
Asigurarea hrnitorilor, adptorilor i Medie 0,5 puncte
cuibarelor conform cerinelor psrilor
Stinghiile Nu exist stinghii propriu - 0 puncte
zise
Numrul de cocoi din efectiv Nu exist cocoi n efectiv 0 puncte
Frontul de acces n exterior 0,64 cm/gin 1 punct
Distana ntre centrul adpostului i 4,195 m 0,5 puncte
deschiderile ctre exterior
Calitatea facilitilor existente n padoc Medie 0,5 puncte
PUNCTAJUL GENERAL/GRUP 3 puncte
LIMITELE DE VARIAIE ALE PUNCTAJULUI GENERAL/GRUP - 2 9 puncte

Aa cum se observ n tabele, cele mai mari punctaje au fost nregistrate de


grupele de factori condiiile de ngrijire i ntreinere respectiv 8,5 puncte din
maxim posibil de 9 i de tipul i caracteristicile pardoselii i suprafeelor n aer
liber respectiv 7 puncte din maxim posibil de 9. Explicaia rezid n aceea c
sistemul alternativ implic att posibiliti mrite de supraveghere a efectivelor i

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de executare a aciunilor sanitare veterinare, ct i accesul la suprafee mari de


padoc i teren nierbat, n care ginile i pot manifesta comportamentul natural.
Tabelul 3
Tabelul de evaluare a grupei a III-a de factori tipul i caracteristicile pardoselii i
suprafeelor n aer liber

Calificativul sau valoarea


Factorii evaluai Notele acordate
msurat/observat
Lungimea stinghiilor 0,354 m/gin 1punct
Calitatea stinghiilor Medie 0,5 puncte
Tipul pardoselii n zona de eliminare
Grtar de lemn 1 punct
a dejeciilor
Calitatea aternutului n zona de
Strat de 15 cm grosime - tala 1 punct
relaxare i scormonit
Starea igienic a aternutului n zona
Aternut uscat, afnat 1,5 puncte
de relaxare i scormonit
Tipul de aternut utilizat pentru
Covoare din plastic 0 puncte
cuibare
Tipul de padoc i starea igienic a Padoc nebetonat, cu stare
1 punct
padocului igienic corespunztoare
Calitatea terenului nierbat Bun 1 punct
PUNCTAJUL GENERAL/GRUP 7 puncte
LIMITELE DE VARIAIE ALE PUNCTAJULUI GENERAL/GRUP - 2,5 9 puncte

Tabelul 4
Tabelul de evaluare a grupei a IV-a de factori iluminarea, calitatea
aerului i zgomotele

Calificativul sau valoarea


Factorii evaluai Notele acordate
msurat/observat
I = 1/41
Iluminarea 1,42 W/m2 0 puncte
Coef. unif. = 0,36
Calitatea aerului Necorespunztoare 0 puncte
Curenii de aer n zona de odihn V = 0,05 m/s 1 punct
Zgomotele din adpost 20 dB 1 punct
Accesul la suprafee exterioare n
255 zile 0,5 puncte
zile/an
Accesul la suprafee exterioare n
2,92 ore/zi 0 puncte
ore/zi
Efectivul pentru care sunt alocate
74,23% 1,5 puncte
umbrare
PUNCTAJUL GENERAL/GRUP 4 puncte
LIMITELE DE VARIAIE ALE PUNCTAJULUI GENERAL/GRUP - 2,5 9 puncte

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Tabelul 5
Tabelul de evaluare a grupei a V-a de factori condiiile de ngrijire i ntreinere

Calificativul sau valoarea


Factorii evaluai Notele acordate
msurat/observat
Starea igienic a hrnitorilor, adptorilor
Foarte bun 1,5 puncte
i cuibarelor
Funcionarea sistemelor i instalaiilor din
Bun 1 punct
dotarea adpostului
Numr mic, moartea s-a
Numrul i starea cadavrelor 0,5 puncte
produs de puin timp
Starea penelor Foarte bun 1,5 puncte
Starea pielii Foarte bun 1,5 puncte
Registrele de sntate Riguros completate 1 punct
Starea de sntate a psrilor Foarte bun 1,5 puncte
PUNCTAJUL GENERAL/GRUP 8,5 puncte
LIMITELE DE VARIAIE ALE PUNCTAJULUI GENERAL/GRUP - 3 9 puncte

Cele mai mici punctaje au fost nregistrate de grupele de factori libertatea


de micare respectiv 1 punct din maxim posibil de 9 i interaciunile sociale
respective 3 puncte din maxim posibil de 9. Aceste punctaje sunt generate de
suprafaa prea mic alocat pe gin i de ponderea redus a suprafeei de relaxare
i scormonit n raport cu suprafaa adpostului.

CONCLUZII

Cercetrile care fac obiectul lucrrii de fa au demonstrat aplicabilitatea


sistemului ANI 35L n condiiile rii noastre. n consecin, sistemul poate fi
propus, n urma verificrilor ulterioare cu rezultate ale evalurilor prin metode de
laborator, spre a reprezenta metoda de evaluare oficial a bunstrii ginilor n
Romnia.
Punctajul final ANI obinut pentru adpostul de gini outoare exploatate n
sistem alternativ a fost de 23,5 puncte; ca urmare bunstarea ginilor se
ncadreaz n categoria bunstrii satisfctoare. Acest punctaj demonstreaz
faptul c sistemul alternativ de exploatare a psrilor a fost conceput avnd ca
principiu satisfacerea necesitilor acestora n condiiile unei productiviti
comparabile sistemului intensiv. De aceea, se recomand aplicarea la scar larg a
acestui sistem de exploatare a ginilor n Romnia, pentru creterea nivelului de
bunstare a efectivelor.

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BIBLIOGRAFIE

1. F. Furnaris, 2004 Evaluarea bunstrii ginilor outoare i a ginilor de


reproducie din rase grele n unele state membre ale Uniunii Europene; Buletinul
Informativ al Societii Medicilor Veterinari n Patologia Aviar i a Animalelor
Mici din Romnia; Volumul 15; Nr. 23
2. F. Furnaris, C. Togan, Elena Mitrnescu, 2005 The assessment of the breeding
hen welfare in a farm from Vrancea country; Lucrri tiinifice Medicin
Veterinar; Seria C; XLVIII; Vol. I; pp. 337-342
3. Teudea V., 2003 Igien veterinar. Adpostirea animalelor. Vol. II, Editura Omega
Print, Bucureti
4. Teudea V., 2005 Bunstarea i protecia animalelor, Editura Omega Print,
Bucureti
5. *** - http://agriculture.de/acms1/conf6/ws5aindexing.htm
6. *** - http://www.bal.bmlf.gv.at/publikationen/hens/ani01h.pdf

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UNELE ASPECTE PRIVIND ACIUNEA AEROIONIZRII


ARTIFICIALE NEGATIVE ASUPRA PROCESULUI
INFECIOS INFLAMATOR CUTANAT INDUS LA
ANIMALELE DE LABORATOR
SOME ASPECTS REGARDING THE NEGATIVE AIRIONIZATION ACTION
ON THE CUTANEOUS INFECTIOUS INFLAMMATION PROSESS INDUCED
AT LABORATORY ANIMALS

Ileana GEORGESCU1, L. ENACHE1,2,


C. FILIPESCU3, G. GEORGESCU2,
I.U. SIMIONCA1, Sanda PENARIU
1
I.N.R.M.F.B. Bucureti
2
U.S.A.M.V. Bucureti
3
I.N.C.D.D.P..B. Victor Babe, Bucureti

The environment is characterized by many parameters, as air


ionization, that is important by its biological effects. In the frame of
artificial ionization application, the action of the air ions could have
therapeutical effects.
The aim. The work intend to present the researches connected with
the influence of the artificial negative ionization on time evolution of the
wound surfaces produced by excoriation and infected with a pure culture of
Staphilococcus aureus at laboratory animals, submited to these
environment conditions.
Materials and methods. It were selected 18 white rats Wistar line
(males) alloted in 2 homogeneousness lots. The generation of the air ions
was made with a Medico type apparatus. The determinations of the air
ionization concentratios were made with ions counter Ebert type (classical
method).
Conclusions. The negative air ions exert a favorable effect on cure
process evolution of wounds. The cure process take places at
concentrations bigger than natural concentrations, beginning with values
over 10,000 negative ions / cm3. The cure process takes place more rapidly
in the case of the negative ions with concentrations of 230,000 ions / cm3.

Key words: airions, cutaneous, infection, macroscopy, microscopy.

Aerul pe care l respir organismele vii se distinge prin o serie de


proprieti fizice, printre care i cele de natur electric, cum este, de exemplu,
ionizarea aerului [1, 9, 10].
Un numr nsemnat de cercetri efectuate de medici, fizicieni, biologi au
evideniat faptul c prezena ionilor atmosferici este una indispensabil vieii,
ntruct aeroionii, n anumite condiii, produc efecte biologice i exercit influene

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directe i indirecte asupra structurilor sau / i funciilor diferitelor nivele de


organizare ale organismelor vii (n funcie de intensitatea ionizrii, polaritatea
ionilor, durata expunerii) [2, 3, 5, 8].
n condiiile unei atmosfere ionizate artificial, cu un regim aeroelectric
crescut, prin expuneri controlate, se pot asigura, relativ uor, condiii de
valorificare a acestui factor de mediu prin inducerea unor influene potenial
favorabile, terapeutice, pentru o serie de afeciuni specifice.
Acest aspect aplicativ a cunoscut o larg rspndire, ndeosebi n occident,
ns, din pcate, aproape de loc n ara noastr [4, 6, 7].

MATERIAL I METOD

Au fost selecionai un numr de 18 obolani albi de laborator linia Wistar


(masculi) repartizai n 2 loturi omogene: lotul 1 - inflamat ionizat (9 animale),
supus aciunii aeroionilor negativi n concentraie de 15.000 ioni / cm; lotul 2 -
inflamat ionizat (9 animale), supus aciunii aeroionilor pozitivi n concentraie de
230.000 ioni / cm.
Animalele au fost anesteziate cu pentobarbital n concentraie de 12 mg /
ml soluie n heparin i apoi, pe partea dorsal a acestora, li s-au provocat o
plag prin grataj care a fost infectat cu o cultur pur de Stafiloccocus aureus n
concentraie etiologic de 109 / ml.
Din ziua a 2-a de la debutul procesului inflamator n cazul lotului 1 i
respectiv din ziua a 4-a pentru lotul 2, animalele au fost supuse cte 4 ore / zi
aciunii aeroionilor, timp de 3 sptmni, fiind crescute n condiii standard de
biobaz.
Generarea ionilor aerieni s-a fcut cu un aparat Medicor (Ungaria) care a
permis obinerea de ioni de ambele polariti (pozitivi sau negativi), ale cror
concentraii se modific proporional cu distana fa de generator.
Determinarea valorilor de concentraie ale ionizrii aerului s-a fcut cu un
contor de ioni de tip Ebert, bazat pe principiul descrcrii unui condensator
electric cilindric (metod clasic).

REZULTATE I DISCUII

Cercetrile privind influena aeroionizrii artificiale s-au concentrat asupra


urmtoarelor aspecte: variaia n timp a suprafeei plgilor induse i a numrului
animalelor vindecate.
Msurarea suprafaei plgilor / lot este prezentat n tabelul 1.

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Tabelul 1
Suprafaa plgii produse / lot la animalele supuse aciunii aeroionizrii de
concentraii diferite

Suprafaa plgii / lot


(cm2)
Tipul de lot
Aeroioni negativi (15.000 Aeroioni negativi (230.000
ioni / cm) ioni / cm)
Inflamat ionizat 3,75 16,333

1. Dup cum rezult i din tab.1, n cazul animalelor supuse aeroionizrii


negative n concentraie de 15.000 ioni / cm (lotul 1), suprafaa plgii produse /
lot a fost de 3,75 cm. Crustele ce se formeaz sunt subiri, sero-hemoragice, cu
margini uor sau marcat decolate.
Evoluia n timp a suprafeei plgii / lot, n cazul animalelor din lotul 1
supuse aciunii aeroionizrii negative n concentraie de 15.000 ioni / cm, se
poate urmri n graficul urmtor (fig.1).

4
3,5
3
Inflam . Ion.
2,5
2
Inflam .
1,5
Neion.
1
0,5 Fig. 1 - Evoluia n timp
0 a suprafeei plgii / lot, n
cazul animalelor din lotul
1 supus aciunii
th y
16 day

y
8t a y
10 da y
5t ay

12 da

da
d
td

aeroionizrii negative
th

th
h

h
1s

concentraie de 15.000
ioni / cm

Evoluia n timp a procesului de vindecare al plgilor provocate la


animalele din lotul 1 a fost urmrit zilnic i este ilustrat prin urmtoarele
imagini macroscopice. Astfel, n ziua 1-a de inflamaie, ionizare din ziua a 2-a,
aspectul macroscopic al plgilor la lotul inflamat ionizat este ilustrat n fig.1.1.
n ziua a 4-a de la inducerea procesului inflamator, ziua a 3-a de
aeroionizare negativ, plgile prezint urmtorul aspect macroscopic, ilustrat n
fig.1.2.
n ziua a 15-a de la inducerea procesului inflamator, ziua a 14-a de
aeroionizare negativ, procesul de reepitelizare prezint urmtorul aspect
macroscopic, ilustrat n fig.1.3.

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n ziua a 20-a de la inducerea procesului inflamator, ziua a 19-a de


aeroionizare negativ, ntreg lotul inflamat ionizat este vindecat, fapt ilustrat i n
fig. 1.4.
Evoluia n timp a procesului de vindecare al plgilor la lotul 1, supus
aciunii aeroionilor negativi n concentraie de 15.000 ioni / cm, poate fi urmrit
n graficul din fig. 2.

Fig. 1.1 Aspectul unor plgi de la lotul Fig. 1.2 Aspectul unor plgi de la lotul
inflamat ziua 1-a, ionizat ziua a 2-a inflamat ziua a 4-a, ionizat ziua a 3-a
(concentraie de 15.000 ioni negativi / cm) (concentraie de 15.000 ioni negativi / cm)

Fig. 1.4 Aspectul unor plgi de la lotul


Fig. 1.3 Aspectul unor plgi de la lotul
inflamat ziua a 20-a, ionizat ziua a 19-a
inflamat ziua a 15-a, ionizat ziua a 14-a
ntreg lotul este vindecat (concentraie de
(concentraie de 15.000 ioni negativi / cm)
15.000 ioni negativi / cm)

10
9
8
7
6
Inflam.
5
Ion.
4
3
2
1
Fig. 2 - Evoluia n timp a
0
procesului de vindecare la lotul
inflamat ionizat supus aciunii
y

y
y

y
da

da

da
da

da

da

aeroionilor negativi
h

th

th

th
4t

6t

8t
10

12

20

concentraie de 15.000 ioni /


cm3

Din experimentul efectuat pe animalele din lotul 1, se observ c procesul


de vindecare este rapid, acesta aprnd nc din ziua a 5-a, cnd se nregistreaz 1
animal vindecat n acest lot. n ziua a 20-a de la inducerea procesului inflamator
ntreg lotul inflamat ionizat este complet vindecat. innd cont de capacitatea de

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regenerare foarte mare a animalelor i de suprafaa prea mic a plgii produse, la


urmtorul lot suprafaa plgii provocate a fost mrit.
Din punct de vedere microscopic, analiznd lamele obinute de la animalele
supuse aciunii aeroionilor negativi n concentraie de 15.000 ioni / cm (lot
inflamat ionizat), se observ la nivelul tegumentului recoltat din locul unde a fost
produs plaga, prezena unui epiderm total regenerat, cu keratin prezent (fig.
1.5), cu o grosime de 0,02592 0,04752 mm, cu limita dermic liniar. n dermul
superficial se constat prezena fasciculelor ngroate de fibre conjunctive
orientate paralel cu suprafaa epidermului i formarea unui esut de granulaie
alctuit dintr-un infiltrat inflamator cronic moderat sau intens, dispus difuz n
zona subiacent epidermului, neocapilare n numr moderat, discret hiperemie,
frecvente fibroblaste active dispuse n jurul capilarelor, unele cu aspecte de
miofibroblaste, cu o intens activitate de sintez a matricei extracelulare i
fibrocite n numr moderat. n dermul profund se observ o textur de fibre
conjunctive groase orientate n toate direciile i un infiltrat inflamator cronic,
discret sau moderat, dispus izolat sau n mici grupuri n special n hipoderm.
La 2 animale din acest lot se remarc existena unei mici zone de
hiperplazie epidermic, a crei grosime este de 0,0648 0,0864 mm (fig. 1.6, 1.7).

Fig. 1.7 - Detaliu la zona cu


Fig. 1.5. - Epiderm regenerat; Fig. 1.6 Zon cu
hiperplazie epidermic: esut
esut de granulaie n dermul hiperplazie epidermic.
de granulaie n dermul
superficial. H&E, 20x H&E, 10x
superficial. H&E, 40x

2. Animalelor supuse aciunii aeroionilor negativi n concentraie de


230.000 ioni / cm (lotul 2), li s-au indus plgi cu suprafee mrite comparativ cu
lotul 1, de 16,333 cm. Crustele ce se formeaz la animalele din lotul 2 sunt mai
mari, mai groase, sero-hemoragice, cu margini nedecolate sau parial decolate.
Evoluia n timp a suprafeei plgii / lot, n cazul animalelor din lotul 2,
supuse aciunii aeroionilor negativi n concentraie de 230.000 ioni / cm, se poate
urmri n graficul din fig. 3.
Evoluia n timp a procesului de vindecare al plgilor provocate la
animalele din lotul 2 fost urmrit zilnic i este ilustrat prin urmtoarele imagini
macroscopice. Astfel, n ziua 1-a de inflamaie, ionizare din ziua a 4-a, aspectul
macroscopic al plgilor la lotul inflamat ionizat este ilustrat n fig.3.1 .
n ziua a 4-a de la inducerea procesului inflamator, ziua 1-a de aeroionizare
negativ, plgile prezint urmtorul aspect macroscopic, ilustrat n fig.3.2.

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n ziua a 19-a de la inducerea procesului inflamator, ziua a 16-a de


aeroionizare negativ, ntreg lot inflamat ionizat este complet vindecat (fig.3.4).

18
16
14
12
10 Inflam.
8 Ion
6
4
2
0
Fig. 3 - Evoluia n timp a suprafeei
plgii / lot, n cazul animalelor din
5t day

th y
th y
th y
y
8t day

th y
10 da
12 da
14 da
19 da
da

lotul 2 supus aciunii aeroionilor


t
h
h
1s

negativi - concentraie de 230.000


ioni / cm

Fig. 3.4 Aspectul unor plgi de la lotul


Fig. 3.1 Aspectul unor plgi de la lotul
inflamat ziua a 19-a, ionizat ziua a 16-a
inflamat ziua 1-a, ionizat ziua a 4-a
ntreg lotul este vindecat (concentraie de
(concentraie de 230.000 ioni negativi / cm)
230.000 ioni negativi / cm)

Evoluia n timp a procesului de vindecare la lotul inflamat ionizat supus


aciunii aeroionilor negativi n concentraie de 230.000 ioni / cm, poate fi
urmrit n graficul din fig. 4. Aa cum rezult i din grafic, n ziua a 13-a apare 1
animal vindecat n lotul inflamat ionizat, iar n 19-a zi ntreg lotul este complet
vindecat.
Din determinrile efectuate pe cele dou loturi se poate observa c, n cazul
animalelor supuse aeroionizrii negative n concentraie de 230.000 ioni / cm, lot
inflamat ionizat, dei suprafaa plgii provocate a fost mrit comparativ cu cea
de la animalele supuse la 15.000 ioni / cm, vindecarea ntregului lot este mai
rapid (a 19-a zi), fa de cea din lotul 1 (a 20-a zi). Rezult din acest context
faptul c, procesul de vindecare se produce la concentraii mai mari dect cele
naturale, ncepnd cu valori de peste 10 000 ioni negativi / cm3, fiind mai rapid n
cazul aeroionilor negativi n concentraie de 230.000 ioni / cm.
Din punct de vedere microscopic, analiznd lamele obinute de la animalele
supuse aciunii aeroionilor negativi n concentraie de 230.000 ioni / cm (lot
inflamat ionizat), se observ la nivelul tegumentului recoltat din locul unde a fost
produs plaga, prezena unui epiderm total regenerat, cu keratin prezent (fig.

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3.5), cu o grosime de 0,02542 0,04968 mm, cu limita dermic liniar. n dermul


superficial se constat prezena fasciculelor ngroate de fibre conjunctive
orientate paralel cu suprafaa epidermului i formarea unui esut de granulaie
alctuit din neocapilare n numr moderat, hiperemie moderat, infiltrat
inflamator cronic moderat dispus difuz n zona subiacent epidermului,
fibroblaste active rare, moderate, sau frecvente, unele cu caractere de
miofibroblaste i frecvente fibrocite. n dermul profund se observ o textur de
fibre conjunctive groase orientate n toate direciile i un infiltrat inflamator
cronic discret sau moderat dispus izolat sau n mici plaje i grupuri n special n
hipoderm.

10
9
8
7
6
Inflam.
5
4 Ion.
3
2
1
0
Fig. 4 - Evoluia n timp a procesului
th y
th y
th y
th y
y
15 da
18 da
14 da

19 da
da

de vindecare la lotul inflamat ionizat


th
13

supus aciunii aeroionilor negativi


concentraie de 230.000 ioni / cm3

Spre deosebire de lotul 1, n cazul lotului 2, se constat la 4 animale


apariia unei zone de hiperplazie epidermic (fig. 3.6, 3.7) cu o grosime de 0,0648
mm (comparabil cu cea obinut la cel 2 animale din lotul 1).

Fig. 3.5 - Epiderm Fig. 3.7 - Detaliu la zona cu


Fig. 3.6 - Zon cu
regenerat; esut de hiperplazie epidermic: esut
hiperplazie epidermic.
granulaie n dermul de granulaie n dermul
H&E, 40x
superficial. H&E, 20x superficial. H&E, 40x

CONCLUZII

Aeroionii negativi exercit un efect favorabil asupra evoluiei procesului de


vindecare a plgilor.

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Procesul de vindecare se produce la concentraii mai mari dect cele


naturale, ncepnd cu valori de peste 10.000 ioni negativi / cm3 .
Procesul de vindecare se produce mai rapid n cazul aciunii aeroionilor
negativi n concentraie de 230.000 ioni / cm, dei suprafaa plgii provocate a
fost mrit, comparativ cu acelai tip de lot supus aciunii aeroionilor negativi cu
concentraie de un ordin de mrime mai mic.

BIBLIOGRAFIE

1. Ardelean I., Barnea M., 1972 - Elemente de biometeorologie medical, Bucureti.


2. Buckalew L.W., Rizzuto A., 1984 - Negative air ion effects on human performance
and physiological condition, Aviation, space and environmental medicine, 55, 8,
p. 731 734.
3. Charry L., 1984 - Biological effects of small ions: a review of findings and methods,
Environmental Research, 34, p. 351 389.
4. Enache L., Andrian C., 1990 Determinri privind influena aeroionizrii asupra
polurii aerului, Lucrri iinifice I.A.N.B., seria E, mbuntiri funciare, vol.
XXXIII, Bucureti, p.35.
5. Enache L., 1999 - Ionizarea aerului i efectele sale biologice, Sntatea plantelor,
nr. 16 (9), Bucureti.
6. Krueger A.P., 1972 - Are air ions biologically significant? A review of a controversial
subject, Int. J. Biometeor., 16, p. 313 322.
7. Inbar O., Rotstein A., Dlin R., Dotan R., Sulman F.G., 1982 - The effect of negative
ion on various physiological functions during work in a hot environment, Int. J.
Biometeor., 26 (2), 153 - 163.
8. Moraru S., Barne M., 1981 Aeroionii, cmpul electric i organismul, Ed. tiinific
i Enciclopedic, Bucureti, 1981.
9. Sulman F.G., 1976 - Health, Weather and Climate, Karger, Baswel, 160pp.
10. Tromp S.W., 1974 Progress in biometeorology, Swets & Zeitlinger B.V.,
Amsterdam.

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CONINUTUL AZOTAILOR I AZOTIILOR N RAIE I


CONCENTRAIA LOR N CARNE I OU LA RAE
THE CONTENT OF NITRATE AND NITRITE IN RATION
AND THEIR CONCENTRATION IN DUCK MEAT AND EGGS

G. IACUB, Viorica GURDI

We made investigations with the goal to determine the pollution with


nitrate and nitrite of meat and eggs from ducks of domestic growth.Though
the forage and the drinking water passed the admitted limit of nitrate, the
poultry received a little quantity of those toxic,nitrate 103 -650 mg /
kilocorpus and nitrite 0,6 4,0 mg / kilocorpus. The daily receive of those
substances did not affect the ducks. Their eggs and meat had organoleptic
propieties. The content of nitrate and nitrite was very reduced, so these
products can be realised without any restrictions.

n ultimii ani s-a micorat considerabil coninutul de azotai n furaje.


Aceast deminuare se datorete ncorporrii n sol a cantitilor mici de
fertilizante (6). ns nectnd la aceasta, problema azotailor rmne actual din
motivul polurii imense a surselor acvatice cu aceste substane (1, 5).

MATERIAL I METOD

Investigaiile au fost efectuate n sectorul particular pe un lot de 83 de rae


(8 masculi i 75 femele), cu vrsta de 10 luni. Psrile sau aflat sub
supravegherea noastr n timp de 9 luni de zile (vara toamna i iarna), cu
determinarea modului de comportare, apetitul i starea clinic, componena raiei,
calitatea sanitar a furajelor i a apei. n furaje i apa potabil o dat n 10 zile s-a
efectuat determinarea coninutului de azotai i azotii, i apoi s-a calculat
coninutul lor n raie pe fiecare sezon aparte. La sfritul fiecrui sezon de la lotul
de rae pentru analiz s-au colectat cte 10 ou i 3 probe de carne din muchii
coapsei.
n furaje, ap potabil, ou i carne coninutul de azotai i azotii s-a
determinat prin metoda electrocolorimetric.

REZULTATELE OBINUTE

n diferite perioade de timp coninutul de azotai i azotii n furaje i apa


potabil variaz considerabil, tabelul 1.
Datele tabelului 1 demonstreaz c n boabele de porumb, gru i amestecul
de concentrate coninutul de azotai i azotii nu depesc limita admis ns
aceast limit admis este depit n rotul de floarea soarelui, verdeaa tocat i
mai cu seam n apa potabil.

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Tabelul 1
Coninutul de azotai i azotii n furaje (mg/kg) i apa potabil (n mg/l)

Limitele de variaie
Specia furajului
Azotai Azotii
Porumb (boabe) 50 - 245 0,90 - 1,75
Gru (boabe) 75 - 195 1,75 - 1,80
Amestec de concentrate 295 - 330 2,50 - 2,75
rot de floarea soarelui 260 - 295 1,20 - 3,50
Verdea tocat 380 - 1900 0,90 - 15,00
Apa potabil 685 - 1300 2,50 - 25,00

Pe parcursul investigaiilor sau efectuat 11 sacrificri de necesitate cu


diagnoza leziunilor traumatice (la cap, gt, aripi, regiunea abdominal i a
membrelor). Traumele au fost cauzate probabil de plasa metalic care avea unele
defecte. Alte careva patologii sau semne de intoxicaie cu azotai i azotii nu sau
constat. Raele aveau un comportament adecvat, faciesul exprim voeciune,
penajul era bine dezvoltat i lucios. Psrile deseori se scldau n bazinul cu ap
sau se odihneau n poziie sterno-abdominal, ele consumau energic hrana i
aveau o productivitate de ou bun.
Furajele utilizate n raia psrilor ct i apa potabil posedau de proprieti
organoleptice bune sau satisfctoare. Coninutul de azotai i azotii n raie i
apa potabil s-a determinat o dat n 10 zile. Datele obinute sau utilizat pentru
calcularea coninutului de azotai i azotii n raie pe fiecare sezon n parte, ct i
cantitatea acestor substane ngerate de rae care revin la 1 kg de greutate
corporal, tabelul 2.
Tabelul 2
Coninutul de azotai i azotii n raie i cantitatea lor ngerat de rae

Coninutul total n raie Greutatea Cantitatea ngerat de


Perioada de
(mg) corporal a rae n mg/kilocorp
investigaie
Azotai Azotii raelor(kg) Azotai Azotii
Vara 269,0 4,80 1,2 224,2 4,00
Toamna 912,0 2,84 1,4 651,5 2,30
Iarna 150,5 0,97 1,5 103,3 0,65

n dependen de varietatea i cantitatea furajelor utilizate coninutul de


azotai n raie constituia n timpul de var 269 mg, toamna 912 i iarna 150,5
mg. Coninutul de azotai era mai mic. Referitor la 1 kg greutate corporal raele
ngerau vara azotai cte 224,2 mg i azotii cte 4,0 mg, n timpul de toamn
corespunztor cte 651,5 i 2,30 i n timpul ernii azotai cte 103,3 mg i azotai
cte 0,65 mg.
Proprietile organoleptice a crnii i a oulor erau bune i foarte bune,
coninutul de azotai i azotii este redat n tabelul 3.

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Tabelul 3
Coninutul de azotai i azotii n carne i ou n mg/kg

Perioada de Carne Ou
investigaie Azotai Azotii Azotai Azotii
Vara 0,6 0,75 1,44 0,51
Toamna 9,8 1,20 0,72 0,43
Iarna 2,0 3,75 1,62 1,10

Datele prezentate n tabelul 3 mrturisesc c n carne i ou se conin


cantiti mici de azotii i azotai. n perioada de var carnea coninea azotai 0,6
mg/kg de product i azotii cte 0,75 mg/kg. Toamna cantitatea de azotai era mai
majorat i constituia 9,8 mg/kg i azotii cte 1,20 mg/kg. n timpul iernei n
carne se conineau azotai cte 2 mg/kg i azotii cte 3,75 mg/kg. Aa dar
coninutul de azotai n carne este mult mai mic ca cantitatea permis care
constituie 200 mg/kg.
n ou cantitatea acestor substane tot este mic. Astfel n timpul de var
cantitatea azotailor constituia 1,44 mg/kg i azotiii 0,51 mg/kg. Toamna oule
conineau azotai cte 0,72 mg/kg i azotii cte 0,43 mg/kg. La finalul
investigaiilor n ou se conineau azotai cte 1,62 mg/kg i azotii cte 1,10
mg/kg n melanj.

DISCUII

Investigaiile s-au efectuat cu scopul de a determina gradul de poluare cu


azotai i azotii a crnii i oulelor obinute de la raele crescute n condiiile
obinuite de cas. Determinarea coninutului de azotai i azotii n furaje
demonstreaz c n majoritatea cazurilor concentraia lor nu depea limita
admis, excepie fiind doar rotul de floarea soarelui i verdeaa tocat n care
limita admis n unele cazuri este depit pn aproape de patru ori. Apa potabil
este impurificat cu aceste substane considerabil. Astfel coninutul de azotai
depete limita admis (45 mg/l) de 15 28 ori i concentraia de azotai
depete limita admis (3,3 mg/l) pn la 75 de ori. Despre poluarea
considerabil a surselor acvatice cu azotai i azotii mrturisesc i sursele literale
(1, 2, 5). n diferite anotimpuri n dependen de cantitatea i asortimentul
furajelor raele ngerau zilnic azotai de la 0,65 pn la 4,0 mg/kilocorp. Aceste
cantiti sunt mici i nu au declanat careva dereglri n starea sntii animalelor
la intoxicaia cronic cu azotai i azotii, ct i despre un coninut redus a acestor
substane n produsele animaliere n literatura de specialitate sunt mai multe
relatri (3, 4 etc).
Investigaiile efectuate pe parcursul a 9 luni de zile au demonstrat c n
carne i ou concentraia de atotai i azotii este foarte mic. Astfel coninutul de
azotai n carne este de 22 77 ori mai mic ca doza permis. Lund n

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consideraie c proprietile organoleptice sunt bune sau foarte bune noi


considerm c aceste produse pot fi realizate fr restricii.

CONCLUZII

Admnistrarea cu hrana azotai de la 103 pn la 651 mg/kilocorp i azotii


de la 0,6 pnla 4 mg/kilocorp nu provoac intoxicaii cu manifestri clinice la
rae.
La intoxicaia cronic a raelor cu azotai de la 103 pn la 651 mg/kilocorp
i azotii de la 0,6 pn la 4 mg/kilocorp, carnea i oule posed propieti
organoleptice bune i foarte bune iar coninutul acestor substane este mic i deci
aceste produse pot fi realizate fr restricii.

BIBLIOGRAFIA

1. Bumbu Ib., Eugenia Tropoel, 1999 - Ocrotirea mediului conform principiului


ecologiei geochimice. Lucrri tiinifice 25 ani de nvmnt superior medical
veterinar n Republica Moldova. Chiinu, p.20.
2. Iacub G., Chiosa A., Slobodeniuc N., Rodidel V., Policiuc I., 2004 - Intensitatea
de poluare cu azotai i azotii a apei potabile din zona central a Codrului.
Simpozion tiinific internaional30 ani de nvmnt superior medical veterinar
din Republica Moldova. Chiinu 1-2 octombrie, p.173 174.
3. Iacub G., Balov S., Moraru I., Ceban R., Bejenaru M., 2004 - Coninutul de azotai
i azotii n raie i concentraia lor n carne la suine. Simpozion tiinific
internaional 30 ani de nvmnt superior medical veterinar din Republica
Moldova. Chiinu 1-2 octombrie, p. 172 173
4. Iacub G., Chiosa A., Balov S., Poliiuc I., 2003 - Coninutul de azotai i azotii n
carne i oule ginelor cu intoxicaie cronic. Simpozion tiinific internaional70
ani ai Universitii agrare de Stat din Moldova Medicin Veterinar, 7 8
octombrie 2003, Chiinu, p. 145.
5. Litjens J.B., Eijkelenburg, F.P. Van., 1987 - Enkele gevallen van nitrietvergiftiging
bij slachtknikenmolderieren. Tijdschr. diergeneesk, 112, N1, 27 28.
6. Rusnac I., Renia E., 1999 - Nivelul nitrailor n nutreuri i profilaxia intoxicaiilor la
animale. Lucrri tiinifice. 25 ani de nvmnt superior medical veterinar n
Republica Moldova. Chiinu, p.32.

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OBSERVATIONS CONCERNING THE AIR QUALITY IN


THE INFLUENCE AREA OF A CHEMICAL UNIT
OBSERVAII PRIVIND CALITATEA AERULUI N ZONA DE INFLUEN
A UNUI COMBINAT PETROCHIMIC

Elena MITRNESCU, Dana TPLOAG,


L. IONI, Violeta SIMION,
Fl. FURNARIS, Cristina DINU

In the area of influence of Arpechim Pitesti Chemical Unit there was


watched the air quality from samples harvested from five checkpoints:
Pitesti, Bradu, Oarja, Prundu and Topoloveni.
In these samples, the following parameters were measured: HCl,
CO, H2CO, NO2, SO2, NH3 and phenols. Analyzes and the results
interpretation were done on the basis of up-to-date standards.
The measurements reach to the following conclusions:
- all the assessed parameters have recorded smaller values than
maximum admitted limits stipulated in STAS 12574/87, in all samples and
all checkpoints;
- the phenols concentrations was constant in all the watched area,
which proves that these chemical compounds are not generated by the
industrial activity of oil processing;
- ammonium and HCl variation of concentrations indicates that
there are others pollutants generators beside the industrial activity of oil
processing.
Industrial activity of Arpechim Pitesti was planed in such manner for
reducing the environmental pollution and its objective is mostly
accomplished.

Pollution became a reality for the most industrial areas and in the condition
of lacking some strong measures of reducing it could produce dangers to the
whole society.
During the campaign for diminishing the pollution, information has got an
essential role. Thus, the pollutant sources must be identified, described and
monitored and also the main pollutants elements, their action, the interdependence
between nixes, the permitted doses, their testing and measuring methods.
The mediating of the pollutant phenomena led to the understanding of the
phenomena and the drafting of their diminishing methods.
The chemical unit Arpechim Piteti, being considered a pollutant source of
the environment, has a permanent activity of monitoring the influence of the oil
industrial processing activity on the environment, conformingly the aspects had in
view by GEMS-Ro. The up-to-date study is a specific one to the environment
protection activity and followed the pollutants elements impact on their air quality

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in the area. The monitoring of the air quality in the influence area of Arpechim
Piteti Chemical Unit is necessary to underline easily and in time of some non-
function during the technologic processes, of some negative phenomena in the
specific substances evolution, the sources and correction modality of these
deficiencies, for avoiding the negative influence on the environment.

MATERIALS AND METHODS

In the influence area of Arpechim Piteti Chemical unit there was followed
the air quality by sampling in five points: Piteti (Environmental Protection
Agency), Bradu, Oarja, Prundu and Topoloveni.
By sampling there were carried out the next chemical parameters: HCl, CO,
SO2, NO2, NH3, phenols and aldehyds. There were chosen the pollutants
mentioned above because they cold have as emit tent the industrial activity of oil
processing and they have major negative effects on plant development.
The determination were carried out conformingly the law stipulated
standards: for NH3 STAS 10712/76; NO2 STAS 10329/75; SO2 STAS
10194/75; HCl STAS 10943/77; fenoli STAS 11027/76 and H2CO STAS
11332/79.
Analyze of the obtained date was made according to the limits stipulated in
12574/85 STAS.

RESULTS AND DISCUSSIONS

The results of the determination regarding the chemical parameters of the


air are shown in table 1.
Table 1
Air quality in the influence area of a Chemical Unit
3
Sampling No. of Average values of the indicators (mg/dm )
point samples HCl H2CO NH3 NO2 SO2 CO fenoli
Piteti
Environmental
199 0,0087 0,0025 0,0022 0,048 0,044 1,576 0,0008
Protection
Agency
Bradu 167 0,013 0,0033 0,028 0,0147 0,040 1,603 0,0011
Oarja 167 0,014 0,0022 0,030 0,0192 0,054 1,582 0,0017
Prundu 178 0,014 0,0014 0,020 0,0142 0,046 1,628 0,0010
Topoloveni 178 0,012 0,0010 0,0193 0,0101 0,049 1,421 0,0010
CMA STAS
- 0,1 0,012 0,1 0,1 0,25 2,0 0,03
12574-87

According to the data in the table, HCl recorded values under the limit of
0,1 mg/dm3 in all samples and all sampling points. Being into the atmosphere in
concentrations under the limits of toxicity, with a maximum admitted

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concentration of 0,1 ppm, HCl has a great effects on plant development, but in
high concentration induce the chlorosis of the leaves and even the appearance of
some necrotic points.
According to the obtained results, the aldehyds didnt exceed the stipulated
limit by STAS 12574/87 in all samples and also all sampling points.
Aldehyds (photochemical oxidant pollutants) results into the air by oil
catalytic transformation in explosive engines and exhausted gases.
Ammonium, even it is not a specific toxic substance for the oil industrial
processing activity and it could react with some specific compounds, constituting
smog. The ammonium content in the atmosphere, being uniform spreaded in the
tested area (table 1) situated more under the maximum admitted limit, along the
whole monitored period.
Nitrogen dioxide recorded values under the maximum admitted limit in all
samples and sampling points.
A nitrogen oxide into the air over the maximum admitted limit provoke
chronic poisoning and leaves scale, and over 10 ppm appear general scales.
Sulph dioxides, according to the tabular data touched concentrations under
the maximum admitted limits by standards in all sampling points. It has a priority
place in atmospheric pollutant group; they could come from different sources, the
petrochemical industry and waste scale and also transportation.
Sulph dioxide concentrations always vary due to the different moving of
the air mass. People and animals, and also plants are sensible to its action,
changing their enzyme systems and essential physiologic processes.
Carbon monoxide, according to the obtained results recorded values under
the maximum admitted limit in all samples.
The amount of phenols in the air were situated at very low levels, more
under the maximum admitted limit in all sampling points.

CONCLUSIONS

The concentration of the carried out parameters were place dat low levels
according to the admitted limit (STAS 12574/87).
The amount of phenols, along the monitored space, shows that they dont
come from the oil industrial processing activity.
The variation of ammonium and hydrogen chloride shows the existence of
other generators than the oil industrial processing activity.
The Chemical Unit oriented itself as the environmental pollution, be more
reduce recording a permanent qualitative evolution of the environment.

REFERENCES

1. Ayres R.U., Ayres, Leslie, 1996 Industrial ecology Towards closing the materials
cicle, Eduard Elgar Publishers, London

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

2. Brown L.R. i colab., 1999 Probleme globale ale omenirii, Editura Tehnic,
Bucureti
3. Marinescu D., 1996 Dreptul mediului nconjurtor, Editura ansa SRL, Bucureti
4. Mitrnescu Elena i col., 2002 Observaii privind poluarea industrial a aerului n
judeul Teleorman, Lucrri tiinifice Med. Vet., Vol XXXV, Timioara, pp. 589-
591
5. Mitrnescu Elena i col., 2002 Observaii privind poluarea industrial a aerului din
zona Trgovite, Revista Romn de Medicin Veterinar, Vol. XII, nr. 1-2, pp.
163-167
6. Negulescu M. i col., 1995 Protecia mediului nconjurtor, Editura Tehnic,
Bucureti
7. Teudea V., 2004 Protecia mediului, Ediia a III-a, Editura Omega Print,
Bucureti
8. Tumanev S., 1989 Calitatea aerului, Editura Tehnic, Bucureti
9. *** STAS 12574/87 Parametri de calitate ai aerului
10. *** Legea nr. 137/1995 Legea proteciei mediului

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OBSERVATIONS CONCERNING COPPER AS A


POLLUTANT ELEMENT IN SOL AND ITS INFLUENCE ON
ANIMAL RAISED IN A LIMITROPH AREA OF A
CHEMICAL UNIT IN THE SOUTH OF ROMANIA
OBSERVAII PRIVIND CUPRUL CA ELEMENT POLUANT DIN SOL I INFLUIENA
ACESTUIA ASUPRA ANIMALELOR DINTR-O ZON LIMITROF UNUI COMBINAT
CHIMIC DIN SUDUL ROMNIEI

Elena MITRNESCU, A. BUTARU,


Dana TPLOAG, L. TUDOR
Carmen PETCU, Elena STAICU

Copper concentration was carried out by samples of soil and organs


(liver, kidney, spleen), sampled from cattle and swine, in form locations:
Isalnita, Almaj, Breasta and Mofleni situated at 2, 6, 8 and 12 km to the
Chemical Unit Isalnita.
Copper analyze was made by atomic absorption spectrophotometry.
Following the researches it can be conclude these:
Copper concentration in soil exceeded the normal and alert limits in
all samples and sampling points, indifferently the distance to the pollutant
source, respective Isalnita Chemical Unit.
The highest copper concentration exceeded the admitted limit by 10
12 times in liver, by 7 19 times in kidney and spleen.
Copper concentration in samples from swine in the influenced area
recorded values over the admitted limit by 6 18 times conformingly the
Health Minister 975/1998.
The exceeding was lower in the sampling points situated at farther
distances to the pollutant source.
The light copper concentrations carried out in the soil and organ
samples, correlated with the clinical and anatomic and pathologic
examination confirm a chronic copper poisoning of animals in the area of
the Chemical Unit.

Chemical industry development led to appearance of some recognized


industrial pollution with implications on health status of environment and
organism.
Such a zone is represented by the village of Isalnita, due to the Chemical
Unit, which pollutes large areas with (pollutant) elements and also copper.
Copper is in the some time an essential and also a toxic element. The
absorbed copper is vehiculated by plasmatic albumins and then fixed in some
organs. The metabolism of copper into the animal organism could be disturbed by
high zinc concentrations, leading to nephrocalcinosis, rib fractures, osteoporosis.

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MATERIALS AND METHODS

The researches had in view the establishing of the copper concentration in


soil, ash and organs, respective liver, spleen and kidneys, sampled in four
locations. Isalnita, Almaj, Breasta and Mofleni, placed at different distances to the
Unit, 2 km, 6 km, 8 km and 12 km. The soil was sampled from 0 20 cm depth.
The organs were sampled form cattle, sheep and swine in the form villages.
The method of emphasizing the copper concentration in samples was the
atomic absorption spectrophotometry.
Analyze of the results was made conformingly the Order of the MAAPPM
184 / 1997 for soil samples and the Order of the Health Minister 975/1998.

RESULTS AND DISCUSSIONS

The results of the analyze regarding the copper concentration in soil are
shown in the table 1.
Table 1
Copper level in the soil samples in Isalnita Chemical Unit area

Sampling Sampling Distance to the Average copper


Sample type
points depht Chemical Unit concentrations (ppm)
Isalnita soil 175,4
0 20 cm 2 km
ash 477,2
Almaj soil 142,1
0 20 cm 6 km
ash 264,2
Breasta soil 112,1
0 20 cm 8 km
ash 164,4
Mofleni soil 69,8
0 20 cm 12 km
ash 88,4
Normal value 20,0
Alert limit - Ord. MAPPM 18 /1997 100,0

Analyzing the data in the table it could notice that the recorded values in
the soil and also ash samples exceeded the normal limits in all sampling points,
indifferently the distance to the Chemical Unit.
The exceeding beside the normal limits was 3.45 8.77 times higher for
soil and 4.42 24 times for ash. The alert limits are exceeded in three sampling
points, respective Isalnita, Almaj, Breasta and Mofleni the values are under these
limits.
The copper concentrations both in soil ash decrease at the some time to the
increasing distance to the Clinical Unit. The average concentration in liver,
kidney and spleen form cattle raised around the Chemical Unit is shown in
table 2.

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Table 2
Average copper concentration in organ samples in cattle in Chemical Unit area

Distance to the Average copper concentration (ppm)


Sampling point
Chemical Unit spleen liver kidney
Isalnita 2 km 54,00 62,42 58,24
Almaj 6 km 51.00 61,00 52,45
Breasta 8 km 44.15 58,75 46,12
Mofleni 12 km 21,14 30,12 21,04
Admitted limit
3,0
Conformingly Health Minister Order 97/1998

Analyzing the data in the table it notice an increasing of the copper


concentration in liver for 10 12 times beside the maximum admitted limit
conformingly the Order of Health Minister Order 975/1998, in spleen 7 -18 times
and in kidneys 7 19,4 times.
This exceeding recorded different values depending on the distance of
sampling to the main pollutant source, the Isalnita Chemical Unit.
In the table 3 there are shown the main concentrations of copper in sheep
samples. As in cattle, in sheep samples it could notice an exceeding of the
admitted limits in all sampling points, an indirect ratio with the distance to the
pollutant source.
Table3
Copper average concentration in organ samples of sheep in Chemical Unit area

Average copper concentration


Distance to the
Sampling point (ppm)
Chemical Unit
spleen liver kidney
Isalnita 2 km 50,12 56,22 52,04
Almaj 6 km 43,22 45,12 42,00
Breasta 8 km 36,12 38,00 31,05
Mofleni 12 km 17,00 19,12 17,82
Admitted limit
3,0
Conformingly Health Minister Order 975 / 1998

Thus, there were carried out the concentrations which exceed the admitted
limit by 6 19 times in liver samples, and by 6 17 times in kidneys and spleen
samples.
Copper is centered upon liver in the central region of his lobule structure
and upon kidneys at the proximal tube cells level.
Bires (1983) [1], following a study, concluded that the copper level in sheep
liver could be asses depending the copper concentration in wool, this increasing
depending on copper liver concentration and Williams (1983) [6] considers the
copper in wool a better indicator for copper liver assessment than plasma.

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Copper concentration in swine organs is shown in table 4.


Table 4
Copper average concentration in organ samples from swine in Chemical Unit area

Distance to the Average copper concentration (ppm)


Sampling point
Chemical Unit spleen liver kidney
Isalnita 2 km 49,.05 59,04 55,0
Almaj 6 km 41,18 53,15 47,92
Breasta 8 km 42,08 47,72 42,14
Mofleni 12 km 16,25 20,02 18,24
Admitted limit
3,0
Conformingly Health Minister Order 97 /1998

Conformingly the tabular date it could notice an exceeding by 20 times in


liver samples in Isalnita, by 18 times in Almaj, 17 times in Breasta and Mofleni.
In kidneys samples, the exceeding beside the admitted limits were 18,6 times
higher in Isalnita, 16,3 times in Almaj, 14 times in Breasta and 6,47 times in
Mofleni. The copper concentration in spleen was over the admitted limit
conformingly the Health Minister Order 975 /1998 by 16,6 times in Isalnita,
14 times in Almaj and Breasta and 6 times in Mofleni.
The highest copper concentrations were established first in liver, then in
kidney and spleen.
These high copper concentrations, met in swine organ samples could be
explained by the fact that the swine food consists of the forages raised in these
influence area.
These high copper concentrations in swine liver didnt allow the iron to be
absorbed in the gastrointestinal tractus, leading to anemia by the lack of iron. The
high copper concentration in soil, correlated to the ones in spleen, liver and
kidneys could demonstrate a chronic copper poisoning.

CONCLUSIONS

Copper concentration in soil and samples exceeded the admitted limits in


all sampling points, indifferently the distance to the Chemical Unit.
Copper concentration in cattle organ samples exceeded the admitted limit
by 10 12 times in liver and 7 19 times in kidney and spleen.
The copper admitted limits in sheep organ samples were exceeded by 6
19 times in liver and 6 17 times in kidney and spleen.
The average copper concentration in swine organ samples exceeds the
admitted values conformingly the Order of Health Minister 975/1998 by 6 18
times, depending on the distance to the pollutant source and also on organ.

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The high copper concentration in soil and organ samples and the clinic
anatomopathologic examination confirm the chronic copper poisoning of the
animals in the Isalnita Chemical Unit.

REFFERENCES

1. Bires J., Vrzgula L., 1983 The content of copper iron, zinc, arsenic, cadmium and
lead in the wool of sheep bred in the vicinity of a copper processing plant. Folia
Vet 27, 2, 103 110
2. Hatch, R. .a., 1979 Chronic copper toxicosis in growing swine. I.A.V.M.A., 6, p
174
3. Elena Mitrnescu i col., 1994 Efectele poluante ale fluorului, cuprului i arsenului
asupra animalelor din zona Valea Clugreasc. Lucrri tiinifice, U.S.B., seria
C, vol XXXVII, (81 86)
4. Elena Mitrnescu i col., 1997 Observaii privind efectele substanelor poluante
asupra bovinelor din jurul Combinatului de ngrminte Chimice Valea
Clugreasc. Lucrri tiinifice, U.S.B., seria C, vol XXXVII, 1994.
5. Elena Mitrnescu, D. Butaru i col. Researches concerning copper pollution
effects on animals in Isalnita Chemical unit area: Animals and Enviroment vol. 2,
290 293, Warsaw, Poland
6. Walliams I.A. i col., 1983 The copper content of wool in relation to breed and the
concentration of copper in the liver and plasma I. Agric. Sci.Camb 100, 505
507.
7. *** Ordinul MS 975/1998 Norme igienico sanitare pentru alimente
8. *** Ordinul MAPPM 184/1997 Proceduri de realizare a bilanurilor de medii.

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CONTROLUL EFICIENEI DERATIZRII


I. IBRU, C. LZRESCU
U.S.A.M.V. Timioara

Rodents are a significant source of Salmonella enteritidis (SE)


exposure for chickens. A single mouse produces 100 droppings a day and
each can contain up to 230.000 SE bacteria. By defecating in feed troughs,
on egg belts and in other areas, rodents can spread infection throughout the
chicken house and contaminate eggs. It appears mice become infected with
SE when exposed to contaminated manure. Placement of contaminated
manure or manure containing infected mice near poultry facilities provides
an additional source for SE exposure. In order to control Salmonella, new
programs must be adopted at all stages of production, processing and
preparation of foods.

Key words: Salmonella enteritidis, rodent control, epidemiology.

Contaminarea cu Salmonella este considerat principala cauz a


toxiinfeciilor alimentare. Cresctorii i comercianii de oua trebuie s colaboreze
pentru a realiza, n continuare, procesul de reducere a ncrcturii bacteriene i
pentru a asigura calitatea produselor specifice.
Pentru a putea controla contaminarea cu Salmonella, trebuie adoptate
programe de supraveghere aplicabile n toate fazele de producie, procesare i
preparare a oulor sau a alimentelor pe baz de ou. La fel n fermele de cretere a
puilor de carne ct i n continuare pe fluxul tehnologic, trebuie s lucreze
mpreuna pentru a se asigura c procesul de reducere a bacteriilor i asigurarea
calitii produselor nu este ntrerupt.
n continuare sunt expui trei factori de risc identificai (din 1995 de ctre
Pennsylvania SE Pilot Study) ca factori de risc semnificativi pentru contaminarea
cu Salmonella enteritidis, SE. Aceste riscuri sunt: contaminarea cu SE a fermelor
avicole; a roztoarelor; a puicuelor pentru reproducie.
n prezent, n fermele acreditate ca libere de SE, foarte multe ou sunt
produse sub autoritatea Programului pentru asigurarea calitii oulor (PACO),
care stpnete cele trei puncte critice incluznd curenia i dezinfecia halelor
pentru puicue i gini outoare ntre fluxuri, controlul roztoarelor i plasarea, n
halele de cretere, doar a puilor indemni de SE.
PACO este un program tip HACCP monitorizat i ajutat de ctre
departamentul pentru agricultur din ara respectiv. Toate persoanele care iau
parte la producerea, procurarea i procesarea alimentelor trebuie s contientizeze
c oule trebuie tratate ca i alimente i nu ca altceva. Totui programele de tip
HACCP nu pot garanta lipsa riscului de contaminare a cojilor de ou cu SE,

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asigurnd consumatorii c n procesul de obinere al oulor se aplica toate


msurile necesare pentru meninerea siguranei oulor.

1. Biosecuritatea factorilor de risc

Biosecuritatea const n folosirea tuturor msurilor de control posibile


pentru a monitoriza rspndirea bolilor cauzate de microorganisme, cum ar fi SE.
Aceste msuri includ: controlul circulaiei oamenilor n fermele avicole; izolarea
pasrilor de animalele sau echipamentele contaminate; controlul roztoarelor i
insectelor, vaccinarea; dezinfecia; pstrarea cureniei n hale.
Scopul biosecuritii este de a menine agenii etiologici la distan fa de
psri. Aceste microorganisme sunt vehiculate dintr-un loc n altul odat cu
gunoiul, praful i penele, prin intermediul aerului i de ctre oameni,
echipamente, vehicule, animale i alte psri. De aceea cel mai mare factor de risc
n introducerea bolilor cauzate de microorganisme sunt oamenii, echipamentele i
animalele contaminate.
Cele mai bune metode de management pentru un bun sistem de
biosecuritate trebuie s cuprind urmtoarele cerine: pentru personal; pentru
echipamente; pentru animale. Succesul programului de biosecuritate depinde n
totalitate de fermier.

2. Curenia i dezinfecia dintre doua populri

Materia organica constituie primul punct de control critic deoarece poate


adposti bacterii, cum ar fi SE. Scopul cureniei i al dezinfeciei este de a reduce
materia organica i bacteriile din mediu, n acest mod se reduce riscul de
contaminare cu SE.
Perioada tranzitorie cuprins ntre doua repopulri permite o buna
curenie, dezinfecie, uscare i inspecia halei. Este de preferat s existe un
interval de dou sptmni ntre dou populri.

3. Controlul roztoarelor

Roztoarele sunt o surs semnificativ de SE pentru pui i constituie al


doilea punct de control critic deoarece un singur oarece poate produce pn la
100 de crotine n fiecare zi, i fiecare crotin poate conine pn la 230000 de
bacterii de SE. Prin defecare n hrnitori, pe banda transportoare a oulor sau n
alte zone, roztoarele pot rspndi infecia n halele de cretere i pot contamina
oule.
Roztoarele se reproduc cu rapiditate n halele avicole, unde mncarea, apa
i adpostul sunt disponibile. Civa oareci intrai ntr-o hal nou pot prolifera
pn la un numr foarte mare (pn la 10000 sau chiar mai muli) pe durata de

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exploatare a unei singure serii de psri. Roztoarele consum furajul, distrug


izolaiile materialelor i slbesc structurile cldirilor prin sparea galeriilor i a
cuiburilor.
Un oarece de casa consuma 5.67 g de furaj de pui zilnic, deci 2000 de
oareci consuma zilnic 11.339 kg de furaj.

3.1. Msuri de prevenire a ptrunderii roztoarelor n interiorul adpostului


Meninerea aleilor betonate din jurul adposturilor ntr-o perfect stare de
curenie pentru a permite observarea roztoarelor de ctre dumanii lor naturali
i pentru a mpiedica accesul lor n interior
Raticidele (momelile sau prfuirile de urmrire) sunt adesea folosite n
programele de control al roztoarelor. Modul de administrare al momelilor este n
funcie de tehnica de preparare a acestora.
Staiile de intoxicare sunt cu succes folosite n multe programe destinate
controlului populaiilor de roztoare. Acestea sunt amplasate de-a lungul aleilor
dintre baterii la intervale de 3-6 metri.
Roztoarele pot fi active pe tot timpul anului n poduri sau pot migra pe
perioada cureniei i a dezinfeciei anexelor. Daca principalul duntor l
constituie obolanul, staiile de intoxicare pot fi amplasate la intervale mai mari n
locuri specifice adposturilor sau locurilor de hrnire, bazate pe tiparul lor de
hrnire. Folosii Formularul de Evaluare al Roztoarelor pentru a le localiza
poziiile.

3.2. Monitorizarea roztoarelor cu ajutorul Indexului pentru Roztoare


Monitorizarea numrului de roztoare dintr-o hal de cretere a puilor este
o parte important a procesului de control al roztoarelor.
Indexul Roztoarelor (IR) este un instrument de nepreuit n monitorizarea
roztoarelor. Indexul folosete, ca metod, att evaluarea vizual ct i un numr
de 12 curse, de capturare a oarecilor vii, pentru aprecierea numrului relativ de
roztoare din hal, ct i pentru verificarea calitii programului de control.
Suplimentar IR poate fi folosit pentru evaluarea unei posibile contaminri cu SE a
unei populaii de psri datorit oarecilor. Un numr crescut de contaminri a
fermelor avicole poate fi pus pe seama oarecilor.
Tabelul 1
Indexul roztoarelor

Numrul de oareci capturai Indexul roztoarelor


0-10 Sczut (1)
11-22 Moderat (2)
Peste 26 Ridicat (3)

Pentru a aprecia populaia de roztoare dintr-o ferm pe baza IR, trebuie


completat Formularul de Evaluare al Roztoarelor, pe timpul zilei cu ajutorul unei

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lanterne. n conformitate cu formularul, se vor amplasa capcane n zona de


circulaie a roztoarelor, cum ar fi de-a lungul pereilor bateriilor, gurilor de
canalizare i n apropierea sistemului de ventilaie. Se vor folosi, n total, 12
capcane. Partea de canalizare aflat dedesubtul zonelor de furajare poate constitui
un loc bun de amplasare a capcanei. Se vor momi capcanele cu aproximativ 28-30
g de furaj pentru psri i apoi vor fi lsate timp de o sptmn, timp n care se
vor verifica de dou ori. Capcanele n care nu au fost capturate roztoare, dup
prima verificare (la 2-5 zile dup amplasare) trebuie s fie mutate n zone unde
prezena acestora este evident. Distanta de mutare trebuie a fi de minimum 5 m.
Scopul sau limita critica este de a menine numrul oarecilor sczut
(IR=1). Dac IR are o valoare mai mare de 1, programul de control ar trebui supus
unei reevaluri. Indexul Roztoarelor ar trebui fcut cel puin o data pe lun
(pentru fiecare populaie de psri). Se va menine o eviden exact a eficienei
fiecrei aciuni.
Inspecia vizuala pe timpul nopii, dup lsarea ntunericului, cu luminile
stinse sau diminea devreme, cu ajutorul unei lanterne, poate fi eficienta pentru
detectarea roztoarelor. Roztoarele pot avea perioade reduse de activitate pe
timpul nopii, de aceea sunt necesare mai multe verificri.

4. Verificarea programului

In mod normal este nevoie de 2 8 sptmni pentru a atinge un IR sczut


in halele care au fost puse in evident populaii de roztoare ridicat sau
moderat. Pentru ca programul de control s fie corect aplicat este necesar
ndeplinirea urmtoarelor cerine:
- ntreinerea corespunztoare a cldirilor;
- efectuarea reparaiilor la nivelul comunicrii cu exteriorul;
- amplasarea staiilor de intoxicare la intervale de 3 15 m n jurul
gurilor de canalizare si a altor zone din adpost;
- complectarea periodic a staiilor de intoxicare cu momeal proaspt;
- verificarea permanent a calitii momelilor folosite;
- evacuarea periodic a dejeciilor din adpost.
Anexa 1
JURNAL DE CONTROL AL ROZATOARELOR

Data Capcane Verificarea Nr. de Substana Produsul


Iniiale IR Momeala
Luna/zi/an amplasate capcanelor roztoare activa utilizat

IR: sczut (1)=1-10 oareci; mediu (2)=11-25 oareci; mare (3)= 26 sau mai muli.

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Anexa 2
FISA DE EVALUARE A ROZATOARELOR
Identificarea locaiei
LOT Luna Ziua Anul

Verificai urmtoarele locuri


Spaii greu Indexul roztoarelor
accesibile,
Nr. Spaiul supus Absena
sau care nu Sczut Moderat Mare
crt. evalurii roztoarelor
au fost (1-5) (6-10) (>10)
verificate
1 Prezena gurilor
n alei i pereii
laterali
2 Prezena gurilor
n pereii frontali
3 Prezena gurilor
lng sistemul de
ventilaie
4 Prezena urmelor
pe ui sau n jurul
acestora
5 Guri sau spaii n
ui
6 Prezena gurilor
n structura
cuibarelor
7 Prezena de guri
i tuneluri n
stratul de izolaie
8 Prezena de guri
i tuneluri n
depozitul de ou
9 Prezena de guri
i tuneluri n
camera de sortare
a oulor
10 Prezena de
ierburi, vegetaie
i arbuti n jurul
adpostului

Populaia de roztoare:
Roztoare prezente ____ oareci ______ obolani _______ ambele
Estimarea populaiei de roztoare zero sau sczuta. medienumeroas ..necunoscut.
Estimarea populaiei de roztoare s-a fcut prin:.inspecie .capturare ambele
Note/comentarii:

Verificare Evaluator

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BIBLIOGRAFIE

1. Baker, R., 1996 Animal Pest management Service, Inc., Chino. 1996
2. Foster, S., Opitz, H.M., Dineen D., 1994 Traning and Reference Manual for
Stapmling of Comercial Pullet and Layer House for Salmonella enteritidis. Univ.
Maine.
3. Henzler D.J., 1993 Determining the number of mice on farms is a dificult task.
Poultry Times
4. Henzler D.J., Opitz, H.M., 1992 The role of mice in the epizootiology of infection
on chiken layer ferms. Avian Diseases nr. 36. p. 625 631
5. Schlosser, W., Henzler D.J., Mason, J., 1995 Salmonelle enteritidis. Pilot project
progress Raport.

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GHIDANCE FOR OPERATORS ON SLURRY


AND MANURE MANAGEMENT
GHID PENTRU OPERATORII N MANAGEMENTUL DEJECIILOR
SOLIDE I LICHIDE

I. IBRU
U.S.A.M.V. Timioara

n prezenta lucrare se prezint un model de calcul al modului de


operare pentru integrarea dejeciilor animalelor. Pornind de la cantitatea
de dejecii, volumul acestora, capacitatea de stocare i n final suprafaa de
care este nevoie pentru a putea fi reintegrarte.

Slurries and solid manures from intensive livestock production are


potentially valuable sources of plant nutrients, but may also be the cause of
pollution. An IPPC installation should be able to demonstrate that the best use is
being made of slurries and manures emanating from the farm and that pollution
risks are being minimized. This can be achieved by preparing and following a
Slurry and Manure Management Plan.
The process of producing a manure management plan has been divided into
eight steps. Where the operator is spreading on his or her own land then s/he is
responsible for all the steps. When the guidance is being used where the manure
or slurry is spread on third party land then Step 1 is the responsibility of the
Operator, who should provide information on the volumes of manure generated
by the installation to the landowner. The landowner/ Planner may then carry out
the remaining steps.
Step 1. Calculate manure and liquid waste production from the installation
Step 2. Allow for other imports of manure effluents or liquid wastes on the
land in the plant.
Step 3. Calculate total manure, effluent and liquid wastes to be
accommodated on the land in the plan.
Step 4. Identify manure spreading areas.
Step 5. Produce a suitable map.
Step 6. Calculate maximum spreading rates.
Step 7. Application methods and timing.
Step 8. Contingency planning

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Calculating manure and liquid waste production


A - calculate total volume of excreta produced by Livestock
Measured production figures can be used, or be taken from Table 1. You
should take account of the number of stock on the unit, and the number of days
the animals are housed to give the total volume produced per year.
If the slurry/manure store is not emptied every year, the base volume
should be taken into account to prevent application rates being higher than
anticipated.
Table 1
Typical volumes of excreta produced

Livestock unit Body weight kg Moisture % Typical vol. L/d


Gilt 90 130 94 10.7
Sow & litter 130 225 94 10.9
Weaner 7 18 90 1.3
Grower 18 35 90 2.7
1000 Laying hens 2200 70 115
1000 broilers $ litter 2200 40 60
1000 broiler breeders 3400 70 217

B- calculate total volume of bedding material used


Measured production figures can be used, or be taken from Table 2.
Table 2
Typical volumes of bedding material used

Tipical Amount used


Livestock Unit Housing System Litter used
per place per year kg
Pigs Pens Straw 102
Wood shavings or
Poultry Deep Litter 1.0
chopped straw of paper
Wood shavings or
Broiler Deep Litter 0.5 per bird per crop
chopped straw of paper

C- calculate the volume of cleaning water used


Measured production figures can be used, or be taken from Table 3.
Table 3
Typical volumes of cleaning water

Livestock unit Cleaning system Range litres Typical litres


Cleaning out pens
Pig after each batch 10 16 24 per batch 18 per batch
pigs per pen
2
Cleaning houses 6.8 per m house
Poultry ( Broilers)
after each crop area per crop

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D- calculate the volume of rainfall to be disposed of


You should measure all areas draining to the slurry/manure store/system, in
order to estimate:
Any roof water not completely separated to the clean drain age system.
Any contaminated yard water.
Areas where livestock have regular access or food.
Areas around feed storage and disposal systems.
Where possible, allowance should also be made for
Any water draining off adjacent land.
Areas where leakage from hoses enter the system.
You should measure the area of the storage system itself, unless the store is
covered and rainwater separated to the clean system.
This total calculated area should be multiplied by the rainfall onto that area
to give the total volume of contaminated rainfall. Rainfall should be calculated on
a minimum of a 5 year return basis (M5) and a 4 month period (1st Novto 28 Feb)
(as a guide the M5 for this period is generally 45% - 50% of the long-term annual
rainfall).

Total Volume produced by the installation = A+ B + C + D

REFERENCES

1. Carton, O.T., Cuddihy, A., Lenehan, J.J., 1994 - Effect of slurry-spreading


technique on silage dry matter yields. Proceedings of the 7th Technical
Consultation on Animal Waste Management. Bad Zwischenahn, Germany.
pp.231-236.
2. Larney, F.J., Olson, B.M., Janzen, H.H., 1993 - Wheat responses to nutrient levels
imposed by simulated erosion and soil amendments. 30th Ann. Alberta Soil
Science Workshop Proc., February 22-24, 1993, Edmonton, AB. pp.131-136.
3. Stevens, R.J., Loughlin, R.J., Frost, J. P., Anderson, R., 1992 - Evaluation of
separation plus acidification with nitric acid and separation plus dilution to make
cattle slurry a balanced efficient fertiliser for grass and silage. Journal of
Agricultural Science, Cambridge, 119: 391-399.
4. Tanaka, D.L., Aase, J.K., 1989 - Influence of topsoil removal and fertilizer
application on spring wheat yields. Soil Sci. Soc. Am. J. 53: 228-232.
5. Teagasc, 1994 - Soil analysis and fertiliser, lime, animal slurry and trace element
recommendations. 36 pp. Teagasc, Johnstown Castle, Co. Wexford.

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IMPLICAIILE ECOMONICE I SANITATR VETERINARE


ALE DEZVOLTRII COMENRULUI CU ANIMALE
I PRODUSE DE ORIGINE ANIMAL
THE ECONOMIC IMPLICATIONS OF GREATER GLOBAL TREDE IN LIVESTOCK
AND PRODUCTS
I. IBRU
U.S.A.M.V. Timioara

GATT (Uruguay Round of the General Agreement on Tarrifs and


Trade), care a fondat World Trade Organisation (WTO), a ajutat la
uniformalizarea presiunilor internaionale pentru liberalizarea comerului,
prin intermediul reducerii tarifelor, restricii cantitative referitoare la
accesul pe pia i reducerea competiiei din partea exporturilor
subvenionate. n acest context, reglementrile tehnice referitoare la
sntatea animalelor i controlul sanitar al importurilor sunt considerate
bariere non-tarif, ceea ce poate conferi posibilitatea unor ri, n special a
rilor cu venituri mici, s profite de un mediu comercial mai liber. Aadar,
aceste reglementri tehnice trebuie supuse unei analize atente, pentru a se
stabili utilitatea lor real. n acelai timp, integrarea grupurilor de naiuni
n zonele de comer liber sau n pieele comune necesit armonizarea
msurilor sanitar veterinare de control al sntii animalelor. n ambele
cazuri, trebuie s se pun n balan ctigurile economice, obinute n
urma comerului liber, cu potenialele costuri ale relaxrii msurilor de
control al bolilor i a msurilor de siguran i de sntate.

Key words: General agreement on tariffs and trade, international trade meat,
sanitary regulations

PRESSURES FOR TRADE LIBERALIZATION

With the development of improved communications of all kinds, recent


decades have seen rapid growth of international trade. It is estimated that the total
value of world trade is, as at 1998, equivalent to roughly one sixth of the
aggregate income of the world; we all live and work in a global economy (1).
However, the growth of international trade has suffered set-backs, such as the
great economic depression of the 1930s. Among the lessons learned was that
removal of trade barriers is likely to encourage economic recovery and growth.
To this end the GATT was established in 1947, with the objective of liberalizing
trade between the 23 countries which were party to the agreement: Since then, the
volume of world trade has increased 16-fold, far more rapidly than aggregate
income; while membership of GATT has risen to 132, with 30 other countries
waiting to join, including Russia and China. The member nations are responsible

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for over 90% of world trade (4). Over a series of rounds of negotiation, tariffs
have been reduced from 40% by value in 1947, to less than 4% in 1998 (1).
The growth of trade in agricultural products has allowed a greater diversity
of diets throughout the world and the establishment of international agricultural
processing industries. Trade in livestock products was largely limited to cross
border movements of animals, until the development of refrigerated transport in
the 1890s. This lead to the creation of major livestock trading routes from the
New World of the Americas, Australia and New Zealand to the Old World of
westem Europe. More recent patterns of livestock trade are discussed below.
Price support for agriculture; including livestock, has long been the norm in
the high income economies, although some countries, notably New Zealand and
Australia, have abandoned the practice. In this context, the Common Agricultural
Policy (CAP) of the European Union (EU) has become notorious for creating
`mountains' of surplus produce (9). Meanwhile, many developing countries,
before and after independence, taxed agricultural exports to finance the general
economic development of the country. Over the 1970s and early 1980s many of
these countries used direct taxes on agriculture, including livestock, and levied
tariffs on exports, whilst others taxed exports indirectly, through the exchange
rate, by maintaining an overvalued domestic currency. The latter results in foreign
exchange being artificially cheap, and imports and exports being undervalued. In
effect, this means that agricultural and other export producers are effectively
taxed by receiving an artificially low price, in the local currency converted at the
official exchange rate. Similarly, the prices of domestic food crops are taxed in
the sense that they are depressed by competition from artificially cheap imports
(10).
These sets of conditions, whereby some countries subsidize agriculture,
while others do not, provide scope for the former group to export surplus produce
at a price below the cost of production in those countries where agriculture is not
protected. This practice, known as dumping, provides unfair competition and a
disincentive to domestic producers.
In the latest round of GATT negotiations; known as the Uruguay Round
(1986-1994), negotiations were extended to cover the problems of protectionism
in the agricultural and textile industries. Under the Agreement on Agriculture,
which was 'signed in April 1994, member countries were committed, by the year
2000, to:
- improve access to imports through the removal of non-tariff barriers
such as quotas the reduction of tariff levels and the agreement of an
upper limit or bound on tariff levels;
- reduce export subsidies by at least.36% (a measure linked with the
anti-dumping code of GATT)
- reduce aggregate measures of support for domestic agricultural
producers by at least 20%.

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These are the main commitments which were agreed, but there are many
detailed exemption clauses and modifications. In particular, less stringent rules
are to apply to developing countries, which have until 2004 to complete
implementation. Furchermore, there is a safeguard clause which allows any
country to apply import restrictions or suspend tariff concessions if products are
imported in such quantities and under such conditions that they may cause severe
harm to domestic producers. However, there is speculation as to the extent that
LICs will be able to benefit from these changes in market access given the
stringency of technical regulations (12).

THE EUROPEAN UNION AND OTHER TRADING BLOCKS

World trade in agricultural products has been greatly affected by the


formation and expansion of regional trading blocks. These range in closeness of
integration, from the free trade area to the economic union, as follows:
- a free trade area has no trade barriers between member states, which
remain independently responsible for trade relationships with the rest
of the world;
- a customs union has no internal trade barriers, but has a mutually
agreed set of tariffs and trade arrangements with the rest of the world;
- a common market, like a customs union, but has freedom of
movement of factors of production between member countries;
- an economic union has even closer monetary, fiscal, social and legal
integration.
The European Common Market became the EU after ratification of the
Maastricht Treaty in 1993. Except for the EU, regional integration has not
generated a high proportion of total trade for member countries. The EU has
succeeded in raising trade within the community to over 60% of all trade, and in
achieving rapid growth of agricultural production. This has been achieved by
imposing quotas and taxes (variable import levies) on imports to the community,
and by subsidising exports. These policies have raised food prices to EU
consumers, created budgetary problems for the community and, it is argued,
encouraged over-intensive levels of agricultural production, with consequential
environmental problems. World market prices for agricultural commodities have
been depressed and destabilized by EU policies, while many non-EU producers
have been denied access to European markets (1, 12). Recent reforms have
reduced the levels of protection for EU producers.
The Lome Agreement (first signed in 1975 and renewed on four occasions),
allows for special, favoured nations agreements with former colonies of the
African, Caribbean and Pacific (ACP) group. Certain commodity exports are
allowed to enter the EU without tariffs, although are generally subject to quotas.
Special arrangements exist for agricultural products, such as sugar and beef

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(Protocol 7), which are also produced within the EU and are therefore govemed
by the CAP (11).
Other Lome provisions include the European Development Fund which
provides aid to ACP countries, and the stabilization of export earnings (STABEX)
scheme. Under the latter scheme, floor arid ceiling prices are agreed for cotton,
cocoa, groundnuts, coffee and sisal, in regular negotiations.. Exporters are
compensated, with cash or credit, when the export price falls below the agreed
floor. Funds are recovered from the CAP exporters, when the ceiling (2).
Over the miid-1990s, the ACP countries supplied 12% of agricultural goods
imported by the EU. However, these agricultural exports accounted for over 40%
of the total ACP exports to the EU.

GLOBAL TRADE IN LIVESTOCK PRODUCTS

Although initially, most world trade and exports from the developing world
consisted of agricultural products, these products have steadily declined as a
proportion of the total. Many' of the LICs which were largely dependent on
agricultural exports, now rely more heavily on exports of oil, other minerals or
manufactured goods (24). This tendency is to be expected, partly because human
food requirements are limited. Thus, the income elasticity of demand for food (the
percentage change in food purchases for a 1 % change in income) is generally
well below unity. In addition, synthetic substitutes have been found or are
emerging for many other non-food agricultural products, such as fibres. These
tendencies have been diagnosed as potential causes of declining terms of trade for
agricultural producers.
Livestock products, by comparison with staple food crops, are relative
luxuries. The value per tonne is much greater than for staple food crops, while the
average consumption per person in the high income countries is several times
higher than in the L1Cs (Table 1).
Table 1
Per capita consumption of livestock products (kg/year) in 1996 (10)

Conrtry Meat Milk


Developing countries in Africa 12.18 33.22
Developing countries in Asia 22.56 34.68
All developing countries 23.92 42:37
All developed countries 75.78 196.64

The income elasticities of demand for meat, dairy products and eggs are
generally close to unity. This means that growth or decline, of incomes has a
substantial effect on the quantities demanded. As populations and per capita
incomes have risen in many parts of the world, so too has the demand for
livestock produce. World production has expanded at a sufficient rate to raise

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average consumption per capita, except possibly in the case of milk (8). However,
growth in production has not occurred equally across countries or products. Pig
and poultry production has expanded much more detail below. At the same time,
trade in pig and poultry meat has grown faster than production of these
commodities (7).
In fact only a small proportion of the livestock production of the world is
marketed internationally. For example, world beef exports in 1996 were 1,67
million tones. This may be explained by the high costs of transporting livestock
and animal products from one country to another, except through cross-border
trade. Generally animal products must be frozen to permit intercontinental
transport. Furthermore, existing health and hygiene regulations limit the extent of
trade. The importation of feed: grains and oilseeds to raise pigs and poultry
domestically may be cheaper than the importation of the equivalent quantity of
meat. Nonetheless, trade in these products greatly exceeds the trade in bovine
meat as a percentage of total production (7,5% for pig meat (of 79.4 million
tonnes produced annually) and 14% for poultry meat (of 56 million tonnes
produced annually).
Until a few years ago, bovine meat (including beef and veal represented the
largest component of world meat trade in terms of both absolute quantity (in
metric tonnes) and value. However, between 1986 and 1996, bovine meat trade
declined in relative terms, growing by 3% annually while total trade in meats
increased by over 5%. Worldwide production stagnated, with 0.4% growth.
Quantities of ovine meat (mutton and lamb) produced (7.3 million metric tonnes)
and traded (approximately 1.2 million tonnes), are much smaller than those of
beef, and have declined by 0.39% annually over the period 1986-1996, although
production has grown by a similar percentage. Pig meat production worldwide is
growing by almost 2% per year, trade in pig meat is growing by 5.4% annually.
Total world poultry meat production has grown rapidly, by more than 5.5%
annually, over the period 1986 1996.

THE BENEFITS AND COSTS OF TRADE

The international enthusiasm and pressure for trade liberalization stem from
the supposed advantages of trade to all participants. These advantages are
illustrated by the theory of Comparative Advantage, originally formulated by the
English politician and economist, David Ricardo in 1817 (5). He argued that if
two countries (or regions, or continents) are considered, each with different
relative costs of production of different commodities, then both countries will
gain from trade, even if one has an `absolute advantage' over the other, i.e. lower
costs of production for all commodities (3).
This is illustrated by the simple hypothetical example given in Table 2.
According to the figures presented here, Europe has an `absolute advantage' over

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Latin America, in that productivity per worker is higher in both meat production
and motor car manufacture. This implies that unit costs (at least for labour) are
lower in Europe. However, while Europe also has a comparative advantage in:
manufacturing motor cars Latin America has a comparative advantage in meat
production This may be expressed in terms of opportunity cost, which is the cost
of producing an item in terms of the alternatives foregone Thus the opportunity
cost of producing a motor car in Europe is only 3.75 tonnes of meat, while in
Latin America it is 5 tonnes of meat. By the same token, the opportunity cost of
meat production, in terms of motor cars foregone, is lower in Latin America than
in Europe (0.2 cars rather than 0.267).
Table 2
Comparative advantage and the benefits from trade

1. Production per worker in Europe and Latin America


Continent Meat (tonnes) Motor cars Tonnes of meat/ car
Europe 15 4 3.75
Latin America 10 2 5,0
2. Effects of transferring six European workers into motor car production
and ten Latin American workers into meat production
Continent Meat (tonnes) Motor cars
Europe -90 +24
Latin America +100 -20
Total production +10 +4
3. Trade occurs with Europe selling 22 motor cars for a price of 4:5 tonnes
of meat each
Continent Meat (tonnes) Motor cars
Europe +9(-90+99) +2(+24-22)
Latin America +1 (+100 -99) +2(-20+22)

REFERENCES

1. Heffernan S., Sinclair P., 1990 Modern international economics. Basil Blackwell,
Oxford, p. 334
2. Leslie J., 1997 -The comparative cost of the-1973 of Newcastle disease in Northern
Ireland under economic and industry conditions of 1973 and 1996. Veterinary
Epidemiology and Economics Research Unit, Department of Agriculture,
University of Reading, p. 50
3. Leslie J., Barozzi J., Otte J., 1997 - The economic implications of a change in FMD
policy a case study in Uruguay. Epidemiol. Sante anim., 31/32 (2).
4. Leslie J.,Upton M., 1999 The economic implication of greater global trade in
livestock and livestock products. Rev. sci. tech. Off. int. Epiz., 18, (2) p. 440
457.
5. MacDiarmid S.C., 1993 - Risk analysts and the importation of animals and animal
products. In Risk analysis, animal health and trade (R.S. Morley, ed.). Reu. sci.
tech. Off. int. Epiz., 12 (4), p. 1055-1092.
6. Paula Lyra T.M. de, 1995 - Sanitary barriers to the beef trade. Agroanalysis, 15 (12),
p. 18- 22.

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7. Raffaelli M., 1995 - Rise and demise of commodity agreements: an investigation into
the breakdown of international commodity agreements. Woodhead Publishing
Limited, Cambridge, p. 240
8. Sere C., Steinfeld H., 1996 - World livestock production systems: current status,
issues and trends. FAO Animal Production and Health Paper, No. 127. FAO,
Rome, p. 82.
9. Suvibank A., 1996 - Capping the CAP? Implementation of the Uruguay Round
Agreement by the European Union. Food Pol., 21 (4/5), p. 393-407.
10. Upton M., 1996 - The economics of tropical farming systems. Cambridge University
Press, Cambridge, p. 374.
11. *** Food and Agriculture Organization (FAO) (1994) - The Uruguay Round Final Act
and its implications for the world ivestock and meat economy. l5th Session of
the Intergovernmental Group on Meat. CCP:ME/94/4, October. FAO, Rome,
p.49.
12. *** Food and Agriculture Organization (FAO) (1996)- Technical barriers to imports of
livestock and meat products from developing countries and progress in their
reduction under the provision of the Uruguay Round Agreements. l6th Session of
the Intergovernmental Group on Meat. FAO, Rome, p.12.

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POSIBILITI DE DETECIE RAPID A


ENTEROVIRUSURILOR I TESCHOVIRUSURILOR SUINE
POSSIBILITYS FOR RAPID DETECTION OF PORCINE ENTEROVIRUSES AND
TESCHOVIRUSES

Adriana Elena ANI, D.C. ANI


U.S.A.M.V. Iai

Porcine picornaviruses comprising at least 23 serotypes grouped


into six species were described as causative agents of neurological
disorders, reproductive failure, and aphthae-like dermal lesions of swine.
Other viruses such as classical swine fever virus (CSFV), African swine
fever virus, pseudorabies virus (PRV), vesicular stomatitis virus, vesicular
exanthema virus, porcine respiratory and reproductive syndrome virus and
porcine parvovirus (PPV) may cause diseases with similar clinical
symptoms. Therefore, rapid and reliable PCR detection of the most frequent
porcine picornaviruses is of interest. A nested RT-PCR protocol was
described suited to detect all known porcine enterovirus and teschoviruses
serotypes using three sets of primer pairs. These primer pairs were
designed to amplify either highly conserved sequences of the 5-
nontranslated region (5-NTR) or the polymerase gene region of the
relevant virus species.

Key words: porcine enterovirus, teschovirus, PCR, nested-PCR

In afar de virusul febrei aftoase i virusul encefalomiocarditei porcine au


fost descrise nc 15 serotipuri de picornavirusuri porcine. Datorit prezenei
genomului ARN i stabilitii virionilor n mediul acid, aceste virusuri au fost
generic denumite enterovirusuri. Iniial au fost descrii agenii cauzali ai
tulburrilor nervoase i de reproducie. Ulterior enterovirusurile porcine au fost
izolate din leziuni cutanate de tip vezicular. Pe baza rezultatelor examenelor
serologice, efectului citopatic (CPE) i proprietilor de replicare pe diferite linii
celulare s-au identificat trei grupe citopatogene: (Zell, 2001, 2003)

Grupa
Genul Virus Serotipuri Boala Simptome
CPE
Enterovirus Enterovirus 2 - Leziuni
porcin dermice de tip
tipul 9, 10 vezicular III

virusul bolii 1 Boala veziculoas Leziuni


veziculoase porcin dermice de tip
porcine vezicular
(SVDV)

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Grupa
Genul Virus Serotipuri Boala Simptome
CPE
Teschovirus Teschovirus 11 Boala Teschen- Tulburri
porcin Talfan Tulburri de nervoase I
(PTV) reproducie Sindromul
Miocardit SMEDI
Tulburri
cardiace
Sapelovirus Enterovirus 1 polioencefalomielit Tulburri II
porcin nervoase
type 8

Pentru depistarea rapid i de certitudine a infeciilor cu aceste virusuri au


fost dezvoltate tehnici de biologie molecular : PCR, nested RT-PCR i mai nou
aprecierea cantitativ a materialului viral folosind real time-PCR. Specificitatea
tehnicilor de biologie molecular se bazeaz pe faptul ca probele cu rezultat
pozitiv la RT-PCR au fost gsite pozitive i n urma examenelor serologice
specifice de depistare a PEV i PTV. (Krumbholz, 2003) In schimb tehnica PCR
este o metod rapid i mai puin laborioas dect izolarea virusului pe culturi
celulare ( prin pasaje succesive) i serotipizarea lui.
Prin PCR sunt amplificate secvenele de material genetic cele mai
conservate n cursul evoluiei filogenetice i ntre serotipuri. In cazul virusurilor
porcine ncadrate n grupul CPE I, II i III, cercetrile au demonstrat c regiunea
5-NTR are cel mai crescut procent de conservabilitate. (Lukashev, 2003) Aceast
secven este format dint-un numr crescut de perechi de baze (neobinuit de
lung)i conine multipli codoni AUG cu rol important n iniierea translaiei
virale. In plus aceste virusuri conin un IRES (internal ribosomal entry site n
locul 5-cap structure. (Zell, 2005)

Reprezentarea schematica a regiunii 5NTR


(Witwer, 2001)

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Structurile conservate ale regiunii 5-NTR ale teschovirusurilor


(Witwer, 2001)

PEV-8 PTV
Omologia celor doua secvene ale regiunii 5-NTR ale enterivirusurilor (PEV-8) i
teschovirusurilor (PTV) (Krumbholz, 2002)

Protocoalele descrise pentru depistarea acestor virusuri porcine prin PCR i


nested RT-PCR difer n ceea ce privete secvena perechilor de primeri.
Perechile de primeri caracteristice fiecrei grupe de citopatogenitate I, II si II
amplific secvene diferite ale 5-NTR, n funcie de regiunea cea mai conservat:
(Zell, 2003)
- Pentru CPE I: secvena de nucleotide 76396 (321 pb)
- Pentru CPE II: secvena de nucleotide 23435 (413 pb)
- Pentru CPE III: secvena de nucleotide 96556 (491 pb)

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Amplificarea teschovirusurilor cu formarea segmentului caracteristic de 321 pb utiliznd


cADN ( obinut prin revers-transcripie) ale serotipurilor PTV (Zell, 2000)

Amplificarea enterovirusurilor serotipul 9 i 10 cu formarea segmentului caracteristic de


491 pb utiliznd cADN ( obinut prin revers-transcripie) ale serotipurilor PEV 9 i 10.
Deasemenea se observ amplificarea segmentelor de cADN ale virusului bolii veziculoase
porcine (Zell, 2000)

Celelalte etape de lucru ale protocolului sunt identice. Deasemenea pentru o


mai mare acuratee a diagnosticului au fost elaborate protocoale de lucru pentru
nested RT-PCR ( n cazul n care cantitatea de material viral din eantion este
redus).

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Datorit faptului c infeciile cu PTV-1, dar nu i cu alte serotipuri de PTV


sunt mai des ntlnite, este necesar serotipizarea. In acest scop a fost formulat o
pereche de primeri ce amplific un segment de 683 pb caracteristic pentru tulpina
PTV-1. Acest fragment corespunde secvenei de nucleotide 1967-2652 i
amplifica regiunea genomicVP3/VP1. (Krumbholz, 2003)
Concluzionnd putem afirma c necesitatea unei tehnici specifice de
detecie a teschovirusurilor i enterovirusurilor porcine se explic prin faptul c
dup epizootiile devastatoare din Europa din prima jumtate a secolului XX,
infeciile cu teschovirusuri au devenit epizootice n multe ri. Dei infeciile cu
aceste virusuri se exteriorizeaz clinic ntr-un procent foarte sczut, prezena unui
agent patogen secundar (bacterii sau virusuri: PRRSV, circovirus porcin)
transform o infecie inaparent intr-o boal clinic.

BIBLIOGRAFIE

1. Kuppeveld F.J.M. van, A.S. de Jong, W.J.G. Melchers, P.H.G.M. Willems, 2005 -
Enterovirus protein 2B po(u)res out the calcium: a viral strategy to survive?;
TRENDS in Microbiology ,Vol.13, nr.2, February, pg. 41-44;
2. Krumbholz A., R. Wurmb, O. Scheck, E. Birch-Hirschfeld, R. Egerer, A. Henke,
P. Wutzler, R. Zell, 2003 - Detection of porcine teschoviruses and enteroviruses
by LightCycler real-time PCR; Journal of Virological Methods nr.113, pg. 5163;
3. Zell R., A. Krumbholz, A. Henke, E. Birch-Hirschfeld, A. Stelzner, M. Doherty, E.
Hoey, M. Dauber, D. Prager, R. Wurm, 2000 - Detection of porcine
enteroviruses by nRTPCR: differentiation of CPE groups IIII with specific
primer sets; Journal of Virological Methods nr. 88, pg. 205218;
4. Krumbholz A., M. Dauber, A. Henke, E. Birch-Hirschfeld, N. J. Knowles, A.
Stelzner, R. Zell, 2002 - Sequencing of Porcine Enterovirus Groups II and III
Reveals Unique Features of Both Virus Groups; Journal of Virology, Vol. 76, nr.
11, June, pg. 58135821;
5. Zell R., M. Dauber, A. Krumbholz, A. Henke, E. Birch-Hirschfeld, A. Stelzner, D.
Prager, R. Wurm, 2001 - Porcine Teschoviruses Comprise at Least Eleven
Distinct Serotypes: Molecular and Evolutionary Aspects; Journal of Virology, Vol.
75, nr. 4, Feb., pg. 16201631;
6. Lukashev A.N., V.A. Lashkevich, O. E. Ivanova, G.A. Koroleva, A.E. Hinkkanen,
J. Ilonen, 2003 - Recombination in Circulating Enteroviruses; Journal of
Virology, Vol. 77, nr. 19, Oct., pg. 1042310431;
7. Doherty M., D. Todd, N. McFerran, E.M. Hoey, 1999 - Sequence analysis of a
porcine enterovirus serotype 1 isolate: relationships with other picornaviruses;
Journal of General Virology, nr. 80, pg. 19291941;
8. Kaku Y., A. Sarai, Y. Murakami, 2001 - Genetic reclassification of porcine
enteroviruses; Journal of General Virology, nr. 82, pg. 417424;
9. Witwer C., S. Rauscher, I.L. Hofacker, P.F. Stadler, 2001 Conserved RNA
secondary structures in Picornaviridae genomes; Nucleic Acids Research, nr. 29,
vol. 24, pg. 5079-5089;
10. Zell R., S. Seitz, A. Henke, T. Munder, P. Wutzler, 2005 - Linkage map of protein
protein interactions of Porcine teschovirus; Journal of General Virology, nr. 86,
pg. 27632768.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

OBSERVATII PRIVIND PREVALENTA UNOR VIROZE


RESPIRATORII LA BOVINE
PROLUSIONS REGARDING THE PREVALENCE OF SOME BOVINES VIRAL
RESPIRATORY DISEASES

D.C. ANI, Adriana Elena ANI


U.S.A.M.V. Iai

Serological screening using blocking-ELISA assay on 114 serums


taken from bovines that havent been vaccinated had positive results for
BVD-MD in a percentage of 43% and for IBR-IPV of 68.42%.
The presence of positive results can be explained by the presence of
the viruses in bovine units (herds) in which the diseases have a subclinical
evolution.

Key words: ELISA ,BVD-MD, IBR-IPV

Infeciile respiratorii ale bovinelor prezint o mare importanta att ca


urmare a etiologiei polifactoriale ct i a susceptibiliti acestora. Dintre toate
microorganismele incriminate n etiologia infeciilor respiratorii ale bovinelor,
virusurile au un rol major, fiindu-le atribuit ceea mai mare importan. Ele
produc infecii primare, cu particulariti epidemiologie i clinico-anatomice, care
de regul sunt complicate cu infecii bacteriene secundare.
Cercetrile efectuate au avut ca obiectiv stabilirea prin examen serologic a
prevalentei infeciei cu virusul BDV-MD si IBR-IPV, n cteva uniti de cretere
a bovinelor din Moldova.

MATERIAL I METOD

Pentru stabilirea prevalentei infeciei cu IBR-IPV si BVDV au fost prelevate


114 probe de snge din trei ferme de bovine situate in Moldova.
Pentru detecia anticorpilor specifici glicoproteinei B a virusului rinotraheitei
bovine si ai proteinei P80 a virusului diareii virale bovine s-a utilizat testul blocking
ELISA .

REZULTATE I DISCUII

Examenul serologic efectuat pe cele 114 probe de ser recoltate de la bovine,


in vederea stabilirii prevalentei infeciei cu virusurile IBR-IPV i BVD-MD au
dus la obinerea urmtoarelor rezultate: (tabelul 1 i 2)

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Tabelul 1
Rezultatele testrilor serologice pentru BVD-MD

Nr.
Unitatea Probe Pozitive Negative Dubioase % Pozitive
crt.
1 F1 20 4 16 0 20 %
2 F2 80 30 45 5 37,5 %
3 F3 14 6 7 1 42,85 %
Total 114 40 68 6 35,08 %

Rezultatele testrilor serologice pentru BVD-MD


80
80

70

60

50
45

40

30
30

20
20 16
14

10 6 7
4 5
0 1
0

F1 F2 F3

Total probe Pozitive Negative Dubioase

Din analiza rezultatelor obinute se constata ca din cele 114 probe


examinate, un numr de 40 (35.08 %) au reacionat pozitiv pentru BVD.
Tabelul 2
Rezultatele testrilor serologice pentru IBR

Nr.
Unitatea Probe Pozitive Negative Dubioase % Pozitive
crt.
1 F1 20 4 16 0 20 %
2 F2 80 60 19 1 75 %
3 F3 14 14 0 0 100 %
Total 114 78 35 1 68,42 %

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Rezultatele testrilor serologice pentru IBR


80
80

70

60
60

50

40

30

20 19
20 16
14 14

10
4
0 1 0 0
0
F1 F2 F3

Total probe Pozitive Negative Dubioase

Procentajul ridicat in cazul probelor pentru IBR-IPV, 78 (68.42%) de


rezultate pozitive din cele 114 ne arata caracterul enzootic al bolii, cu mare
difuzibilitate in focar.
Din tabelul 1 i tabelul 2 se observ c la toate probele examinate s-au
depistat animale seropozitive pentru IBR-IPV si BVD-MD. Procentele reactiilor
pozitive difer de la o unitate la alta. Trebuie precizat c rezultatele reflect
situaia existent n anul 2005.
Rezultatele obinute in urma testrilor serologice a probelor se arat c cele
dou viroze evolueaz concomitent. In marea lor majoritate probele au fost
declarate pozitive att la BVD-MD ct i la IBR-IPV.

CONCLUZII

1. Ancheta serologica prin intermediul metodei blocking ELISA a stabilit


seroprevalenta infeciilor cu BVD-MD si IBR-IPV la trei ferme din Moldova,
prevalenta ceea mai ridicata nregistrndu-se in cazul IBR-IPV.
2. Prezenta reaciilor pozitive, colaborata cu lipsa fenomenelor clinice in
efectiv se poate explica prin existena unor infecii latente. Acestea sub aciunea
unor factorii abiotici sau biotici favorizani pot declana apariia boli in efectiv.
3. Rezultatele obinute sugereaz c este necesar o extindere a
investigaiilor serologice in ct mai multe ferme pentru a depista animalele cu
forme subclinice.

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BIBLIOGRAFIE

1. www.oie.int, 2004 - Manual of Diagnostic Tests and Vaccines for Terrestrial Animals
2. Merk Veterinary Manual, seventh edition;
3. Perianu T. i colab., 2005 Boli infecioase ale animalelor, Viroze, Vol II, Ed.
UniversitasXXI, Iai;
4. Ctan N., 1998 Infeciile respiratorii ale tineretului bovin, Editura Brumar,
Timisoara;
5. Carp-Crare M., 2001 Microbiologie veterinar. Virusologie, Casa de editur
Venus, Iai;
6. Sandvik T., 1999 - Laboratory diagnostic investigations for bovine viral diarrhoea
virus infections in cattle, Veterinary Microbiology
7. Thiry E., 2002 Maladies Virales des Ruminants, Collection Virologie clinique, Ed.
Le Point Veterinaire;

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Universitatea de tiine Agricole i Medicin Veterinar Iai

INFECIILE LATENTE CU BHV-1:


MECANISM I IMPORTAN
THE LATENCY OF BHV-1: MECHANISM AND IMPORTANCE

D.C. ANI, Adriana Elena ANI


U.S.A.M.V. Iai

Following primary infection of the eye, oral cavity, and/or nasal


cavity, bovine herpesvirus 1 (BHV-1) establishes latency in trigeminal
ganglionic (TG) neurons. Virus reactivation occur after natural or
corticosteroid-induced stress. Reactivation from latency occurs
sporadically, resulting in virus shedding and transmission to uninfected
cattle. Infection of calves with BHV-1 leads to infiltration of lymphocytes in
TG and expression of IFN- (interferon-gamma), even in latently infected
calves. During latency, virus antigen and nucleic acid positive non-neural
cells were occasionally detected in TG suggesting there is a low level of
spontaneous reactivation.

Key words: herpesvirus bovin -1, IBR-IPV, stare de laten.

Herpesvirusul bovin-1 (BHV-1) este agent cauzal al rinotraheitei


infecioase bovine (IBR), vulvovaginitei pustuloase (IPV) i balanopostitei
infecioase (IBP) .
Infeciile latente reprezint una din cele mai importante probleme asociate
infeciei bovinelor cu virusul IBR-IPV: herpesvirusul bovin 1. Att tulpinile
virulente circulante cat si cele vaccinale atenuate pot produce infecii latente ce
pot fi reactivate de diferii stimuli precum utilizarea glucocorticoizilor (ex:
dexametazona).
Influena latentei BHV-1 asupra epidemiologiei infeciei: animalele rmn
purttoare latente dup prima infecie cu tulpini virulente; tulpinile atenuate
rmn latente dup vaccinare; nu se poate controla diseminarea tulpinilor
atenuate; vaccinarea nu previne fixarea (infecia) n stare latent a tulpinilor
virulente; poate fi posibil recombinarea ntre o tulpin atenuat i una circulant
pe un animal cu infecie latent infectat cu ambele tulpini; statusul imun al
animalului cu infecie latent influeneaz nivelul reexcreiei.
Infeciile latente permit persistena BHV-1 ntr-o unitate fr a fi necesar
ptrunderea unui virus exogen.
n urma infeciei primare cu BHV-1 boala poate evolua sub form de
rinotraheit sau conjunctivit, urmat de generalizarea infeciei pe diferite ci:

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

- viremie;
- pe cale neuronal;
- prin intermediul punilor intercelulare evitnd lichidul extracelular
(aa se explic diseminarea virusului de la o celul la alta n prezena titrurilor
crescute de anticorpi) = principala cale de diseminare dup reactivare.
Herpesvirusul bovin 1 (BHV-1) face parte din subfamilia
Alphaherpesvirinae i prezint unele proprieti biologice asemntoare cu ale
herpes simplex virus tipul 1 i 2 (HSV-1 i HSV-2) (Jones et al., 1990; Jones,
1998). BHV-1 prezint o perioada de laten destul de lung n ganglionii
sistemului nervos periferic nainte de replicarea iniial n mucoasa epitelial. De
cele mai multe ori virusul se reactiveaz i este eliminat dup un stres natural sau
indus cu hormoni corticoizi. Cercetrile recente au demonstrat latena virusului
rinotraheitei infecioase bovine (BHV-1) n neuronii senzitivi din ganglionul
trigemen (TG) i in centrii germinativi ai tonsilelor faringiene. (Winkler et al.,
2000)
Reacia caracteristic la nivelul ganglionilor trigemeni n urma infeciei este
reprezentat de infiltraie limfocitar. Cercetrile au artat ca limfocitele T
controleaz infecia cu herpesvirus-1 n ganglionii senzitivi (Simmons et al.,
1992) i s-a putut observa ca celulele neuronale infectate cu antigen viral sunt
nconjurate de celule care elaboreaz IL-2, IL-6, IL-10 i IFN-. (Shimeld et al.,
1997) In infecia acut a bovinelor cu BHV-1, zone de infiltraie limfocitar pot fi
rapid detectate. Numeroase studii au dus la concluzia c celulele mediate ale
rspunsului imun persist n ganglionii trigemeni la bovinele cu infecii latente
produse de BHV-1. (Winkler et al., 2002). Persistena celulelor imune n TG este
dat de expresia proteinei virale ntr-un neuron sau celul satelit. Examinarea
atent a neuronilor TG a bovinelor cu infecii latente a artat c ntr-un neuron se
gsete o cantitate crescut de material viral transcris, proteine virale i AND viral
replicativ. (Feldman et al., 2002) Aceti neuroni care exprim material viral
transcris i proteine sunt nconjurai de zone de infiltraie limfocitar i au fost
descrii ca fiind locul de reactivare molecular spontan (spontaneous
molecular reactivation) a infeciei. (Feldman et al., 2002) Persistena sistemului
imun n TG pe parcursul infeciilor latente are rolul de reactivare a ciclului
infecios. Limfocitele T care produc interferon- joac un rol important n
prevenirea reactivrii infeciei latente. (Liu et al., 2001) In plus limfocitele
citotoxice pot inhiba difuzarea virusului n TG prin inducerea apoptozei. IL-2
este o citotoxin neuroprotectiv elaborat de marea majoritate a neuronilor i a
celulelor gliale (Lovato L. 2003) care stimuleaz expansiunea limfocitelor T i
secreia de interferon- (Jones C., 2006)

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Infiltraie cu celule mononucleare la bovine cu infecie BHV-1. Col. HEA, x 250 (la 6 zile
post infecie i la 14 zile postinfecie) (Jones C., 2006)

In urma infeciei pe cale nazal sau ocular cu BHV-1 are loc transportul
virusului n TG i instalarea infeciei latente. In cazul unei infecii acute n TG se
observ exprimarea genelor virale urmate de instalarea strii de laten. (Jones,
2006) Astfel are loc o inflamaie cronic infiltrativ cu limfocite n ganglionii
trigemeni ca urmare a infeciei cu BHV-1. Exprimarea citokinelor inflamatorii n
timpul infeciilor latente este rezultatul exprimrii reduse a genelor virale sau a
nivelului redus de reactivri spontane. (Shimeld et al., 1997).
Leziunea caracteristic: infiltraii focale interstiiale cu celule
mononucleare poate fi detectat dup 7 zile de la infecie. In cazul infeciei latente
se pot observa discrete zone cu infiltraii cu mononucleare ca i la scurt timp de la
debutul tratamentului cu dexametazon: dup 6 i 24 de ore. Dup 48 de ore de la
administrarea de dexametazon zonele de infiltraie sunt mult mai largi i mai
greu de localizat. Secreia de citokine de ctre celulele inflamatorii (IL-2, IFN- i
IFN-) determin o inhibiie a infeciei productive n TG, deci pot media infecia
cu BHV-1, dirijnd instalarea i reactivarea strii de laten. (Winkler M., 2002)
Datorit faptului c s-a putut detecta in infeciile latente prezena acidului
nucleic si a expresiei genelor virale in celulele non-neurale (celule satelit) din TG
s-a concluzionat ca nivele sczute de reactivri spontane promoveaz prezena de
limfocite pe perioada fazei de laten. Replicarea viral in acest numr sczut de
celule nu a dus la excreia de virus pe care nazala sau ocular. (C. Jones, 2006)
Este bine cunoscut faptul ca glucocorticoizii au capacitatea de a reactiva o
infecie latent cu BHV-1, avnd efect imunosupresiv i putnd cauza apoptoza
limfocitelor. (Schynts F., 2003). innd cont c secreia de citokine are rol in
represia viral, odat cu moartea celulelor care secret cytokine este favorizat
reactivarea infeciei. n schimb aciunea glucocorticoizilor de reactivare a infeciei
este mai sczut in neuroni n comparaie cu cea din celulele non-neurale (satelit).
De asemenea aciunea citokinelor secretate de celulele infamatorii ce nconjoar
in cazul infeciei latente celulele infectate este mult mai puternica fa de neuroni
dect fa de celulele satelit (ce conin ARN replicativ i expresii ale genelor
virale). (Lovato L., 2003)

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Infiltraie cu celule mononucleare n TG la bovine infectate cu BHV-1 Col. HEA, x250


A - infecie acuta: 7 zile de la infecie; B - infecie latent: 60 zile de la infecie;
C - infecie latent: 6 ore de la administrarea dexametazonei; D - infecie latent: 24 ore de
la administrarea dexametazonei; E - infecie latent 48 ore de la administrarea
dexametazonei; F - infiltraie cu celule mononucleare; (Winkler M., 2002)

Pn n prezent este demonstrat c BHV-1 are capacitate de a infecta


limfocitele T CD4+ n cazul unei infecii acute (Winkler et al., 1999) i c
limfocitele conin ADN viral n infeciile latente. (Winkler et al., 2002)
Concluzionnd se poate afirma c dei rspunsul imun este pronunat,
virusul nu este niciodat ndeprtat (distrus complet) din organismul infectat ci el
rmne intr-o stare de laten prelungit, putnd fi oricnd reactivat sub aciunea
anumitor factori. Pentru depistarea i eradicarea bolii este strict necesar
identificarea i eliminarea obligatorie a animalelor cu infecii latente, care sunt de
fapt purttori sntoi de virus. Din pcate utilizarea vaccinurilor este doar o
soluie temporar i limitat. Astfel inct aceast strategie de eradicare a
purttorilor de virus presupune costuri ridicate pentru depistarea serologic.

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Att controlul ct i eradicarea IBR nu sunt dect inte. Este nevoie de


mbuntirea a metodelor de lucru: vaccinuri mai bune i mai bune posibiliti de
testare.

BIBLIOGRAFIE

1. Jones, C., 1998 - Alphaherpesvirus latency: its role in disease and survival of the
virus in nature. Adv. Virus Res., nr. 51, pg. 81133.
2. Jones, C., Delhon, G., Bratanich, A., Kutish, G., Rock, D., 1990 - Analysis of the
transcriptional promoter which regulates the latency-related transcript of bovine
herpesvirus 1. J. Virol. nr. 64, pg. 11641170.
3. Winkler, M.T., Schang, L.S., Doster, A., Holt, T., Jones, C., 2000 - Analysis of
cyclins in trigeminal ganglia of calves infected with bovine herpesvirus-1. J. Gen.
Virol. nr. 81 (Pt. 12), pg. 29932998.
4. Winkler, M.T.C., Doster, A., Jones, C., 2000 - Persistence and reactivation of
bovine herpesvirus-1 (BHV-1) in tonsils of latently infected cattle. Persistence
and reactivation of bovine herpesvirus 1 in the tonsil of latently infected calves. J
Virol. nr. 74, pg. 53375346.
5. Winkler, M.T., Doster, A., Jones, C., 1999 - Bovine herpesvirus 1 can infect CD4(+)
T lymphocytes and induce programmed cell death during acute infection of
cattle. J. Virol. nr.73, pg. 86578668.
6. Winkler, M.T., Doster, A., Sur, J.H., Jones, C., 2002 - Analysis of bovine trigeminal
ganglia following infection with bovine herpesvirus 1. Vet. Microbiol. Nr. 86, pg
139155.
7. Simmons, A., Tscharke, D., Speck, P., 1992 - The role of imune mechanisms in
control of herpes simplex virus infection of the peripheral nervous system. Curr.
Top. Microbiol. Immunol. nr. 179, pg. 3156.
8. Shimeld, C., Whiteland, J.L.,Williams, N.A., Easty, D.L., Hill, T.J., 1997 - Cytokine
production in the nervous system of mice during acute and latent infection with
herpes simplex virus type 1. J. Gen. Virol.nr. 78, pg. 33173325.
9. Feldman, L.T., Ellison, A.R., Voytek, C.C., Yang, L., Krause, P., Margolis, T.P.,
2002 - Spontaneous molecular reactivation of herpes simplex virus type 1
latency in mice. Proc. Natl. Acad. Sci. U.S.A. nr. 99, pg. 978983.
10. Liu, T., Khanna, K.M., Carriere, B.N., Hendricks, R.L., 2001 - Gamma interferon
can prevent herpes simplex virus type 1 reactivation from latency in sensory
neurons. J. Virol. nr. 75, pg.1117811184.
11. Lovato, L., Inman, M., Henderson, G., Doster, A., Jones, C., 2003 - Infection of
cattle with a bovine herpesvirus 1 (BHV-1) strain that contains a mutation in the
latency related gene leads to increased apoptosis in trigeminal ganglia during the
transition from acute infection to latency. J. Virol. nr. 77, pg. 48484857.
12. Jones C., V. Geiser, G. Henderson, Y. Jiang, F. Meyer, S. Perez, Y. Zhang, 2006 -
Functional analysis of bovine herpesvirus 1 (BHV-1) genes expressed during
latency, Veterinary Microbiology nr.113, pg. 199210.
13. Ackermann M., M. Engels, 2006 - Pro and contra IBR-eradication, Veterinary
Microbiology, nr.113, pg. 293302
14. Schynts F., F. Meurens, B. Detry, A. Vanderplasschen, E. Thiry, 2003 - Rise and
survival of bovine herpesvirus 1 recombinants after primary infection and
reactivation from latency, Journal of Virology, Dec. 2003, Vol. 77, nr. 23 pg.
1253512542

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EPIDEMIOLOGY OF EXPOSURES WITH HIGH RISK FOR


RABIES INFECTION AND RABIES POSTEXPOSURE
PROPHYLAXIS
P. CAZACU, M. MARES,
U.S.A.M.V. Iai
DANIELA CAZACU
Center Medical, Trgu Frumos, Iai

The aim of this study is to present the epidemiology of exposures


with high risk for rabies infection (EHRRI) occurred in human population
from Iasi county (Romania) between 1999 2003 and to emphasized the
features of rabies postexposure prophylaxis.
Information about EHRRI from St. Parascheva Infectious Diseases
Hospital database was summarized for a period of five years. The incidence
of EHRRI for county, various age and sex groups was calculated. Also,
animal species involved in these EHRRI, location of wound and surgical
procedures performed, features of rabies postexposure prophylaxis and
period of hospital-stay were evaluated.
In the period mentioned above, 925 individuals were involved in
EHRRI. From age-groups, the highest incidence was in children less than 5
year-old (60/100,000 person-years) and from sex-groups, the highest
incidence occurred in males (126/100,000 person-years). From 925
involved in EHRRI 920 received rabies postexposure prophylaxis (43.91%
rabies serum, 23.36% rabies vaccine, 32.60% both and 0.13% no data
about the administered product). The total duration of hospital-stay was
4,093 days and the number of individuals that required surgical procedures
(sutures, amputations and reconstructive techniques) was 154 (16.64%).
From 975 injuries (bites and scratches), 24.53% from lesions occurred on
head, face or neck, 9.96% on trunk, back or buttocks, 50.43% on arm, hand
or finger and 13.92% on leg, foot or toe. The exposures were due to dogs
66.16%, cats 15%, horses 8.32%, rats 4.21%, goats 1.83%, foxes
1.08%, other species 3.67%.
Interventions such as education in order to prevent animal bite,
vaccination of carnivores against rabies, children surveillance, reduction of
stray-dogs and rodents control must be instituted, enhanced or better
enforced in communities.

Key words: epidemiology, exposures with high risk for rabies infection,
rabies postexposure prophylaxis, Iai-Romania.

Veterinarians are an important source of information for the public and


their role in prevention of zoonoses is unanimously recognized.

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Postexposure prophylaxis (PEP) is given in response to a perceived need


for rabies prophylaxis in the event of a bite, scratch, or other human exposure to a
potentially or definitively rabid animal. Often, PEP is administered without
confirmation of rabies exposure (to err on the side of caution) even though
guidelines for treatment exist. Rabies PEP is expensive and not without potential
adverse effects. Local reactions at the injection site or systemic problems, such as
muscle aches, fever, or vomiting may develop.
Animal bite or exposure reports may be one source of information about
PEP.
The need for PEP is determined by obtaining an accurate history and
decisions to administer PEP often are made in counsel with a rabies exposure
expert.
Surveillance data are important for identifying conditions that require
further investigation and for providing sensible solutions from imperfect data to
facilitate public health action (4).
The aim of this study is to present the epidemiology of exposures with
high risk for rabies infection (EHRRI) occurred in human population from Iasi
county (Romania) between 1999 2003 and to emphasized the features of rabies
postexposure prophylaxis.

MATERIALS AND METHODS

Informations about EHRRI from St. Parascheva Infectious Diseases


Hospital database was summarized for a period of five years (1999-2003). Copies
of paper exposures with high risk for rabies infection reports were reviewed for
Iasi county, and fields common to most reports were entered into a computerized
database.The incidence of EHRRI for county, various age and sex groups was
calculated. Also, animal species involved in these EHRRI, location of wound and
surgical procedures performed, features of rabies postexposure prophylaxis and
period of hospital-stay were evaluated.
EHRRI consist in any penetration of the skin by teeth and resultant
contamination of the wound with potentially infectious saliva, scratches,
abrasions, open wounds, mucous membranes with saliva (lick) or other potentially
infectious materials such as brain tissue from a rabid suspect animal (i.e.
veterinarians, veterinary students, certain laboratory workers, spelunkers) and
ingestion of meet belonging to contaminated animals or rarely in transplantation,
casual contact such as petting a rabid suspect animal (1,2).
Victim information was entered into a computerized database which
included residence, age, location of wound and surgical procedures performed,
and postexposure treatment.
Incidences (numbers of new cases/population at risk) of EHRRI were
calculated, using 1999 2003 populations. Data were analyzed, using a statistical
software program (Excel for Windows Professional).

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RESULTS

Exposures with high risk for rabies infection (EHRRI) report were colected
from Iasi county (Romania) between 1999 2003. In this period have been
registered 925 EHRRI, with an average incidence of 22.36 exposures/100,000
person-years, from which only 920 individuals have received PEP. The highest
incidence of EHRRI occurred in 2001 (i.e. 29.80 exposures/ 100,000 person-
years) - see Table 1.
Table 1
Annual incidence of EHRRI from Iasi county (Romania) between 1999 2003

Rate per 100,000


Year Number of exposures Population
person-years
1999 109 833 388 13.07
2000 185 836 751 22.1
2001 251 842 126 29.8
2002 181 805 330 22.47
2003 199 816 003 24.38

From data registered at territorial laboratory for rabies diagnosis, in Iasi


county (Romania) no cases of rabies in humans were confirmed between 1999
2003, but 27 cases of animal rabies were identified.
Table 2
Age-specific rates of reported animal exposures with high risk for rabies infection
(EHRRI)

Age-group Number of exposures Population 1999-2003 Rate per


(years) 100,000 person-
1999-2003 Mean 1999-2003 Mean years
<5 158 31.6 259 878 51 975.6 60.79
5 to 9 119 23.8 252 219 50 443.8 47.18
10 to 14 102 20.4 332 643 66 528.6 30.66
15 to 19 71 14.2 355 306 710 612.0 1.99
20 to 24 50 10.0 411 035 82 207.0 12.16
25 to 29 49 9.8 355 878 71 175.6 13.76
30 to 34 46 9.2 333 942 66 788.4 13.77
35 to 39 24 4.8 223 076 44 615.2 10.75
40 to 44 31 6.2 259 715 51 943.0 11.93
45 to 49 53 10.6 276 654 55 330.8 19.15
50 to 54 41 8.2 233 799 46 759.8 17.53
55 to 59 31 6.2 162 244 32 448.8 19.10
> 59 150 30.0 677 191 135 438.2 22.15
Total 925 185 4 133 580 826 719.6 22.36

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Age-specific rates for EHRRI were also calculated. Adjusting for the
population size of each age group, children < 5 years old have the highest
incidence of EHRRI - 60.79/ 100,000 person-years (Table 2). Incidence of
EHRRI is maximal in individuals 20 year-old and less (1.69 fold greater than that
observed in the group of individuals with age >59 and 2.68 fold greater than in
group of individuals with ages between 20-59 years, the difference between the
studied groups being statistically significant (p< 0. 001 ), Table 2.
From the table below we can noticed that the incidence of EHRRI in men is
greater than that observed in women (p < 0. 05), see Table 3.
Table 3
Sex-specific rates of reported animal EHRRI.
Date from Iasi county (Romania) between 1999 2003

Number of Population 1999- Rate per 100,000


Sex-group
exposures 2003 (Mean) person-years
Men 518 409407.2 126.52
Women 407 417312.4 97.52

975 injuries (bites, scrapes) have been registered. Most victims have
suffered injuries on the arm, hand, or finger (50.43 %); 24. 53 % were bitten on
the head, neck, or face; 13. 92 % on the leg, foot, or toe; and 9. 96 % on the trunk,
back, or buttocks. 1.12 % of reports lacked information on wound or injuries
location. Children <10 years old were more likely to be exposure on injuries on
the head, neck, or face compared with people >10 years old (P < 0.05, Table 4).
Table 4
Frequency of animal injuries by wound location and victim age
Date from Iasi county (Romania) between 1999 2003 *

Number of injuries (%)


Age group
(years) Head, face, Trunk, back, Arm, hand,
Leg, foot, toe
neck buttocks finger
<5 118 (70.6 %) 10 (5.9%) 31 (18.5%) 6 (3.5%)
5 to 9 71 (54.1 %) 16 (12.2%) 33 (25.1%) 11 (8.3%)
10 to 14 34 (31.7 %) 17 (15.8%) 43 (40.1%) 11 (10.2%)
15 to 19 19 (25.3 %) 6 (8%) 41 (54.6%) 9 (12%)
20 to 24 12 (22.2 %) 8 (14.8%) 30 (55.5%) 4 (7.4%)
25 to 29 6 (11.5 %) 9 (17.3%) 30 (57.6%) 7 (13.4%)
30 to 34 7 (15.9 %) 2 (4.5%) 27 (61.3%) 8 (18.1%)
35 to 39 5 (17.2 %) 5 (17.2%) 17 (58.6%) 2 (6.8%)
40 to 44 10 (29.4 %) 4 (11.7%) 14 (41.1%) 6 (17.6%)
45 to 49 12 (21.8 %) 2 (3.6%) 30 (54.5%) 10 (18.1%)
50 to 54 4 ( 9 %) 3 (6.8%) 27 (61.3%) 8 (18.1%)
55 to 59 2 (6.4 %) 1 (3.2%) 21 (67.7%) 7 (22.5%)
> 59 6 (3.9 %) 13 (8.5%) 91 (59.8%) 38 (25%)
Mean % 24.53% 9.96% 50.43% 13.92%
* 11 of 975 total reports injuries for Iasi County, 1999-2003, were missing wound location

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Most animal EHRRI reports indicated the species of the involved animal.
More than 66 % were dogs, 15 % cats, 8.32 % horses, 4.21 % rats, 1.83 % goats,
1.08 % foxes, 0.97 % ferret, and 2.7 % other species (Table 5).
Table 5
Frequency of animal species involved in EHRRI,
from Iasi conty (Romania) between 1999-2003

Number of
Other
exposures Dog Cat Horse Rat Goat Fox Ferret
species
1999-2000
925 612 136 77 39 17 10 9 25
% 66.16 15 8.32 4.21 1,83 1.08 0.97 2.7

The incidence of dog exposures was 14.8/100,000 person-years (18/ 1,000


person-years, USA Overall, 2001) and the incidence of cat exposures was
3.29/100,000 person-years.
From 925 involved in EHRRI 920 received rabies postexposure
prophylaxis (43.91 % rabies serum, 23.36 % rabies vaccine, 32.60 % both and the
number of individuals that required surgical procedures (sutures, amputations and
reconstructive techniques) was 154 (16.64%), and 0.13 % no data about the
administered product, Table 6).
Table 6
Frequency of rabies post-exposure prophylaxis (PEP) or / and surgery.
Date from Iasi county (Romania) between 1999 2003 *

Number of rabies post-exposure


Type of PEP and treatment
prophylaxis and surgery (%)
Antirabies serum 404 (43.91%)
Antirabies vaccine 215 (23.36%)
Antirabies serum and Antirabies vaccine 300 (32.60%)
Surgery 154 (16.64%)
* 5 of 925 total reports didn't have PEP and one of 920 no information regarding PEP

The total duration of hospital-stay was 4,093 days (mean/ person 4.48 days
hospital-stay/ person).

DISCUSSION

Exposures with high risk for rabies infection (EHRRI) are an important
community health problem. Serious injuries, particularly to children, and the need
for rabies PEP are major consequences. Because of the combined efforts of local
and state health departments, animal control offices, and the veterinary
community, rabies in humans is rare and results primarily from sporadic
exposures to the foxes strain of rabies. Continued surveillance and rabies control

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is still necessary to protect public health, but new issues such as the relative costs
and benefits associated with various control measures have emerged .
Reducing the need for PEP includes reducing rabies in wild animal
populations through such methods as bait vaccination or foxes. Rabies in foxes is
the most important reservoir for human and domestic animal exposure in
Romania and Iasi county, and foxes exposures resulted in at least 10 PEP
treatments between 1999-2003. Means by which the number of stray-dogs and
rabies in stray-dogs may be reduced include euthanasia, neutering, and
vaccination of these animals, eventually their adoption.
Another way to reduce the need for rabies PEP is to increase vaccination
rates for pet (owned) animals (3, 4). A final way to reduce the need for PEP is by
educating children and adults about bite prevention and dog behavioral Dogs were
responsible for most reported EHRRI in our study.
The care and management of dogs and the interface between dogs and
people appear to be major issues, especially for households with or near children.
Reducing rabies PEP also includes minimizing unnecessary administration
of PEP.
When dealing with exposures to livestock, rodents, lagomorphs, physicians
were asked to consult public health officials and reminded that bites from,
hamsters, guinea pigs, gerbils, rats, mice, rabbits and hares almosts never required
anti-rabies treatment. Administration of PEP to person EHRRI by animals
currently vaccinated against rabies is not recommended.
The estimated incidence of PEP was 22.25/ 100,000 person-years. Other
studies indicate an incidence of 4.6/ 100,000 person-years (in Pennsylvania state,
USA, 1995). So, in Iasi county it is of 4.83 more likely (4).
Costs of PEP were still high, respectively 65 Euro/PEP/case, and 15 Euro/
day of hospitalization. The costs for EHRRI increase through the sick leaves, the
surgical reconstruction procedures, or then patients remain with a body of psychic
handicap after aggression or mutilation.

CONCLUSIONS

The highest incidence of EHRRI occurred in 2001 (i.e. 29.80 exposures/


100,000 person-years).
Adjusting for the population size of each age group, children < 5 years old
have the highest incidence of EHRRI - 60.79/ 100,000 person-years.
Incidence of EHRRI in men is greater than that observed in women
(p<0. 05).
975 injuries (bites, scrapes) have been registered. Most victims have
suffered injuries on the arm, hand, or finger (50.43 %).
Children <10 years old were more likely to be exposure on injuries on the
head, neck, or face in comparison with people >10 years old (p < 0.05).

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The incidence of dog exposures was 14.8/100.000 person-years (the highest


incidence).
From 925 involved in EHRRI 920 received rabies postexposure
prophylaxis. The estimated incidence of PEP was 22.25/ 100,000 person-years.
Continued surveillance and rabies control is still necessary to protect public
health, but new issues such as the relative costs and benefits associated with
various control measures have emerged.

REFERENCES

1. Corey L., 2001 Rabies virus and other rhabdoviruses, in Harrisons principles of
internal medicine 15th edition, McGraw-Hill.
2. Fishbein B.D., Bernard W.K., 1995 Rabies Virus, in Mandell, Douglas and
Bennetts -Principles and Practice of Infections Diseases, 4th editions, Churchill
Livingstone Inc.
3. Krebs W.J., Noll R.H., Rupprecht E.C., Childs E.J., 2002 Rabies surveillance in
the United States during. JAVMA, Vol. 221, No. 12, December 15, p. 1690-170.
4. Moore A.D., Sischo M.W., Hunter A., Miles T., 2000 Animal bite epidemiology
and surveillance for rabies postexposure prophylaxis. JAVMA, Vol. 217, No. 2,
July 15, p. 190-194.
5. Overall L.K., Love M., 2001 Dog bites to humans-demography, epidemiology,
injury, and risk. JAVMA, Vol. 218, No. 12, June 15, p. 1923-1933.
6. The National Association of State Public Health Veterinarias Committee (USA)
Compendium of Animal Rabies Prevention and Control, 2003. JAVMA, Vol. 222,
No. 2, January 15, 2003, p. 156-161.

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PCR DETECTION OF V. PARAHAEMOLYTICUS WITH


TLH GENE PRIMERS
Gh. BRDAN
U.S.A.M.V. Iai

The study describes the optimization of the parameters and the test
of PCR to establish a specific and significant PCR method for detection tlh
gene of Vibrio parahaemolyticus. The results show that the use of PCR
clearly increases the effectiveness of the method by detecting this bacterium
of soiled seafood. PCR-based developed improves considerably the speed
and the sensitivity of detection of V.p. Through the development of the tools
of molecular biology, the microbiological safety of the seafood for human
consumption can be increased.

Key worlds: detection, V. parahaemolyticus, PCR, electrophoresis

INTRODUCTION

Polymerase Chain Reaction (PCR) is a molecular biology technique that


allows for quick replication of DNA (4). The principle of the reaction of gene
amplification is to recopy a segment of DNA or RNA in many specimens, thanks
to a DNA thermostable polymerase extracted from Thermus aquaticus (Taq
polymerase) and with two oligonucleotidic starters which frame the amplified
segment. A certain number of requirements are however related to the food
diagnosis with knowing: need for distinguishing the bacteria alive from the dead
bacteria, very weak test specimens used in these tests, many inhibitors present in
food (2). These requirements prohibit any use on line of the genetic tests.
Vibrio parahaemolyticus (V.p.) is frequently isolated in coastal water and
the seafood in temperate zones throughout the world (3). It is the most frequent
cause of the food borne disease in the areas where many residents eat believed
fish. It is a halophytic Gram-negative bacterium usually associated with seafood.
A high incidence of this pathogen undoubtedly originates from the frequent
consumption of marine food. Some foods implicated in the US are crab, shrimp,
and lobster, which unlike fish in Japan, typically were cooked before eating (5).
Mishandling practices, such as improper refrigeration, insufficient cooking, cross-
contamination, or recontamination are suspected in these outbreaks (3). Clinical
manifestations have included diarrhea, abdominal cramps, nausea, vomiting,
headache, fever, and chills, with incubation periods ranging from 4 to 96 hours
(1).
The objective of this study was to develop and evaluate PCR assays to
confirm and characterize V. parahaemolyticus using primers for tlh gene.

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MATERIALS AND METHODS

Bacterial strains and microbiological media. Assay specificity was


determined for the V. parahaemolyticus strains (ATCC 17.802). Bacterial strains
were grown overnight at 37C in alkaline peptone water with 3% salt (APW3%).
V. parahaemolyticus was cultured in T1N3 agar plates (10% tryptone, 3% NaCl)
at 37C overnight. Other vibrio strains were grown and maintained as follows V.
cholerae, V. alginolyticus, V metschnikovii and V. vulnificus.
DNA purification for PCR optimization. Total genomic DNA from V.
parahaemolyticus and other bacterial strains was purified as described by Wizard
Genomic DNA purification kit (Promega Corporation, Madison, WI, SUA). The
DNA concentration was measured at 260/280 nm.
Selection of target and oligonucleotide primer sequences. A segment
of the tlh gene of the V. parahaemolyticus (GenBank accession no. M36437) was
used as the PCR target for specific detection. Potential primer sequences were
analyzed for specificity by comparison with known gene sequences by using the
GenBank database with the BLAST search program and by using the nucleotide
sequence analysis developed in this study. It is located between bp 904 and 927
of the DNA segment with the sequence: 5'-AAA GCG GAT TAT GCA GAA GCA
C-3' and 5'-CGT-TCA-TCT-CAA-GCA-CTT-TCG-CA-3'. The primers used in this
study were custom synthesized by Eurogentec Liege, Belgium.
Optimization of PCR and specificity of oligonucleotide primers.
Purified genomic DNA from V. parahaemolyticus was used for the optimization of
PCR amplification. All PCR reagents were obtained from Amersham Pharmacia
Biotech, Holland). PCRs were performed at a final volume of 20 l. Each reaction
mixture contained 2 l of buffer 10x; 2.0 mM MgCl2; 0.5 mM (each) dATP, dCTP,
dGTP, and dTTP; 0.2 mM primers; 0.2 l Taq polymerase; 18l water ultrapure
and 2 l of extracted DNA. All PCR amplifications were performed in a DNA
thermal cycler Mastercycler Gradient Eppendorf (VWR, Leuven, Belgium), with
the following PCR cycling parameters: initial denaturation at 94C for 5 min,
followed by 40 cycles of amplification. Each amplification cycle consisted upon
the denaturation at 94C for 30 s, primer annealing at 58C for 30 s, and primer
extension at 72C for 30 s.
Detection of amplified DNA. All PCR-amplified DNAs were separated at a
constant voltage of 5 V/cm in 1% (wt/vol) agarose (Eurogentec, Lige, Belgique)
with 1 TAE (per liter, 40 mM TrisCl [pH 8.0], 1.18 ml of acetic acid, 2 mM Na2-
EDTA). The separated DNA in the gel was stained with 2% ethidium bromide per
ml and visualized on a U.V. illuminator (Bio-Rad, Nazareth, Belgium). The
amplified DNA bands were photographed with a Kodak 1D (Thermolabsystems,
Brussels, Belgium)
Specificity of detection. The specificities of the oligonucleotide primers
and the target DNA segment, tlh, for the detection of V. parahaemolyticus were
tested by PCR amplification of the purified genomic DNA of all the strains listed
above by using the optimum PCR conditions and cycling parameters described.
Detection of V. parahaemolyticus in contaminated raws. V.
parahaemolyticus was grown overnight in T1N3 plates at 37C, inoculated into

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fresh APW3% broth, and grown at 37C to approximately 4,2 108 cells x ml-1 as
determined by viable plate counting. Cells were 10-fold serially diluted to
extinction in 25 g of sterile raws in 225 ml APW3%. These dilutions were
incubated at 37C for 18 hours. After this interval the bacterial count indicated
3x108 ufc/ml.. DNA was isolated from 1 ml of the sample and then diluted 10-fold
and PCR were performed.

RESULTS AND DISCUTIONS

Specificities of oligonucleotide primers and optimization of PCR and


cycling parameters. The oligonucleotide primers, L-tlh and R-tlh, amplified the
targeted tlh gene segment of all V. parahaemolyticus isolates. PCR amplification
was determined to be optimal at an annealing temperature of 58C, by a
succession of assays. Decreased intensities of the amplified product in an agar gel
were observed with gradient of dilutions DNA sample. At an annealing
temperature of 56C, nonspecific amplicons in addition to the targeted tlh gene
fragment were observed. PCR-amplified DNA bands was an expected molecular
size of 130 bp (Fig. 1).

Fig. 1. Electrophoresis migration pattern of PCR assay detects the tlh gene. Reference
strains of V. parahaemolyticus; lane 1 to 6, reference strains at T+, respectively 41;
comparison of selected strains of other vibrios (45-48)

The present study provides the premise for developing a rapid, real-time
fluorogenic detection of this pathogen in marine foodstuff and coastal waters.
However, further optimization of the PCR amplification protocol and
hybridization reactions may be necessary to achieve these objectives.
The minimum level of detection was the third decimal dilution, respectively
3x10 V. parahaemolyticus cells x ml-1 in bivalvular mollusks (Fig. 2).
4

Further study or modification of the commercially available DNA


purification kits is necessary to achieve the necessary sensitivity for the detection
of this pathogen in shellfish and shellfish-growing waters (2). This PCR approach
can potentially be used to monitor with high specificity and well within the

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required range of sensitivity the occurrence and distribution of this newly


emerged pathogenic V. parahaemolyticus strain in coastal, marine, and ship
ballast waters (3). Early detection will help increase seafood safety and decrease
the risk of infectious outbreaks caused by this pathogen.

Fig. 2 Agarose gel electrophoresis of


PCR-amplified tlh DNA segments from
V. parahaemolyticus in contamination
raw at diluted 10-fold.
m: DNA size markers.

CONCLUSIONS

The PCR protocol for the tlh gene of V. parahaemolyticus is specific and
rapid, the detection can be achieved in 4 h, but the quantification is limited.
Early detection of V. parahaemolyticus will help increase seafood safety
and decrease the risk of infectious outbreaks caused by this pathogen.

REFERENCES

1. Bacteriological Analytical Manual http://www.cfsan.fda.gov/~ebam/bam-9.html


2. Bej, A.K., D.P. Patterson, C.W. Brasher, M.C.L. Vickery, D.D. Jones, C. Kaysner, 1999 -
Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using
multiplex PCR amplification of tl, tdh and trh. J.Microbiol.Meth; 36: 215225.
3. DePaola, A., L.H. Hopkins, J.T. Peeler, B. Wentz, R.M. McPhearson, 1990 - Incidence of
Vibrio parahaemolyticus in U.S. coastal waters and oysters. Appl.Environ.Microbiol; 56:
22992302.
4. McPherson M.J., Moller S.C., 2000 - PCR The basics of PCR, Bios Scientific Publishers Ltd.,
2000, 89-119.
5. Myers M., Panicker G., Bej A.K., 2003 - PCR Detection of a Newly Emerged Pandemic Vibrio
parahaemolyticus O3:K6 Pathogen in Pure Cultures and Seeded Waters from the Gulf of
Mexico. Appl Environ Microbiol.April; 69(4): 21942200

ACKNOWLEDGEMENTS

I would like to thank Dr. Bernard China and Prof. Georges Daube of Liege
University, Belgium, for their assistance in project and use the PCR protocol.

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HYBRIDIZATION DNA-DNA SPECIESSPECIFY METHOD


ON NYLON FILTERS FOR DETECTION
OF V. PARAHAEMOLYTICUS TLH GENE
Gh. BRDAN
U.S.A.M.V. Iai

Methods of detection based on probes DNA was developed and was


characterized like alternatives to the long and less specific conventional
protocols bacteriological. Our objective was to develop and evaluate a
genetic method for specific detection of V parahaemolyticus in bacterial
colonies of insulation. Hybridization DNA-DNA of the evaluation of the
method was carried out for the gene tlh of a standard stock (ATCC) of V.
parahaemolyticus. Probes marked with digoxigenin were prepared by the
PCR. Analyses of hybridization of colony were carried out using the nylon
filters. The lyses of colony, hybridization, and the immunologic parameters
of detection were optimized for the identification of the biotypes of target.
The experimental process presented here can be filled in 2 day. The
protocol was conceived specifically to identify V parahaemolyticus in the
seafood. This methodology allows also the confirmation or the classification
of the suspect colonies on filters when a culture was not recovered.

Key words: V. parahaemolyticus, detection, DNA hybridation

INTRODUCTION

The principle of molecular hybridization is based on the capacity which


two DNA strands (the target ADN and the ADN probe) have to pair in a specific
way by their complementary bases to form a duplex. The technology of the
probes uses this principle to connect the target chains of the micro-organism to
test with called marked chain probes. Enormous progress was made in the no
radioactive systems of marking: enzymatic marking, by fluorescence and
luminescence (2).
Vibrio parahaemolyticus is a halophilic bacterium often found within
shellfish and in estuarine waters. Among a number of pathogenic vibrio species, it
is commonly associated with oyster-borne gastroenteritis (4). In general, in
optimum temperatures, V. parahaemolyticus grows more rapidly than other food
poisoning-causing bacteria, and negligence in temperature control during food
processing and storage is the major cause of poisoning by this pathogen.
The most commonly accepted method for detection of V. parahaemolyticus
in food is described in the FDA Bacteriological Analytical Manual (BAM) (1). It
is established 1x104 V. parahaemolyticus cells g-1 as a level that warrants
regulatory action for ready-to-eat seafood products (3). This includes pathogenic

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(Kanagawa phenomenon-positive) and non-pathogenic (Kanagawa phenomenon-


negative) strains, both of which must be enumerated in these foods.
The purpose of this study was to evaluate a non-isotopic species-specific
probe for thermolabile haemolysin (tlh) gene that is safe to handle and that can be
used to reduce the time and effort necessary to verify the identity of V.
parahaemolyticus from seafood products.

MATERIALS AND METHODS

The study was made in Laboratory of microbiology of foodstuffs of animal


origin (LMDA) at the University of Liege, Belgium.
Bacterial strains and culture preparation. Three V.p. strains (ATCC
17.802) and other vibrio species strains (V. alginolyticus , V. cholerae O1, V.
metschnikovii , V. vulnificus) were provided by LMDA culture collections. Many
samples were analyzed by the FDA-BAM method (1) using a procedure with
enrichment in APW3% salt and incubated at 37C for 18h, and isolation on
thiosulphate-citrate-bile salts-sucrose (TCBS) agar plates. The sucrose-negative
(green) colonies were selected from each plate for screening using API-20E
diagnostic strips (BioMerieux, Paris, France). Unless otherwise stated, all media
and equipment were sterilized by autoclaving for 15 min at 121C.
Vibrio parahaemolyticus gene probe. The nucleotide base sequence for
the probe was from bases 904927 of the V. parahaemolyticus tlh gene
(accession number M36437) was synthetized by Eurogentec Liege, Belgium: 5'-X
AAA GCG GAT TAT GCA GAA GCA C X-3' and 5'-CGT-TCA-TCT-CAA-GCA-
CTT-TCG-CA-3'. The polymerase chain reactions (PCR) were realized with
Mastercycler Gradient Eppendorf (VWR, Leuven, Belgium). Probe and filter
preparation, hybridization and colorimetric detection were done according to
Wizard Genomic DNA purification kit (Promega Corporation, Madison, WI, USA)
and QIAquik PCR Purification Kit (Qiagen GmbH, Hilden, Germany).
Detection of Vibrio parahaemolyticus by tlh gene probe. The bacteria
grown in agar broth were plated on T1N3 agar (1% tryptone, 3% NaCl and 2%
agar) using a tips of 1 l for each spot, with incubation at 37C, 18h (Fig. 1). The
colonies (Fig. 2) were transferred to nylon filters HybondTM-N+ (Amersham Life
Science, Buckinghamshire, England) and treated with alkaline solution for DNA
denaturizing and cross-link with the filter. After pre-hybridization, the filters were
probed with the Digoxigenine-labelled tlh probe by using DIG High Prime DNA
Labeling and DetectionStarter Kit for color detection with NBT/BCIP (Roche
Applied Science, Applied Science, Penzberg, Germany).

RESULTS AND DISCUSSIONS

We prepared and verified a probe of DNA of V.p. by the PCR with starters
for the gene tl (24 pairs of bases: 904-927). The probe was marked with
Digoxignine. One carefully respected the indications of the manufacturer for the

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hybridization of the ADN fixed on a nylon membrane, followed of an


immunological detection of this hybridization (Fig. 3).

Fig 1. Disposition of the Fig. 2 Agar T1N3 : bacteria Fig. 3. Nylon filter after
bacteria to transfer them on colonies after 18h at 370C immunologic detection: the
filter arrow indicates the colored
spot corresponding to V.p.

As the method was not used usually in the laboratory, we had many the
difficulties by setting up it. After several tests, our conclusion is that this method
is very delicate and tiresome.
The difficulty of the technique is also related to the use of the many
solutions. As sequence in time, the detection of V.p. takes approximately two
days, almost half of detection by traditional bacteriology, and with a reduced
reliability. One of the significant points of the technique consists in sowing, on an
agar, small spots of solution of enrichment. According to the bacterial
concentration, these drops can have 1, 2, 5 or 10 l; the sense of smell of the
technician played an important part in the choosing of the volume necessary. As
V.p. is a mobile bacterium, after a few hours of incubation, the bacterial colonies
become increasingly large and can join, making difficult the interpretation of the
results.
To obtain a probe of ADN it is necessary to carry out a traditional PCR, to
purify the product of PCR, to concentrate it and to mark it with Digoxygenine.
The checking of marking still took a few hours. The process of hybridization is
the most significant stage of the method. The temperature of 42C seemed to be
highest with the fixed probe, but it was far from that of 58C used during the
hybridization of the PCR. In addition, one attended with an incomplete
hybridization, in spite of the continuous agitation of the membrane in the solution.
One met several times the phenomenon of precipitation of the solution of
hybridization, another limiting factor of the method. It should be added the fact
that the use of the gene tlh does not make it possible to detect the pathogenic
stocks of V. parahaemolyticus, i.e. that this method is not useful for the detection
of routine.
The team of Gooch (2) compared two known methods for detection of V.
parahaemolyticus by hybridization of ADN species-specific, direct on membrane
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nylon, by using two probes for the gene tlh: one marked with alkaline and
different phosphatase and one with the digoxignine. Their conclusion was that
hybridization is an easy and fast method. Recently, another American team
proposed to carry out hybridizations with probes for the genes tlh and tdh marked
with alkaline phosphatase, in order to detect the total number of V
parahaemolyticus and to identify the pathogenic stocks (5).

CONCLUSIONS

These results suggest that the probes are possibly for detection of the V.
parahaemolyticus tlh gene.
Specific hybridization DNA on the nylon membrane is a difficult method to
realize in the detection of V parahaemolyticus in routine process and comprises
many difficulties.
Through this method it is impossible to establish a quantification of the
pathogenic bacteria in a food product.

REFERENCES

1. Elliot, E.L., Kaysner, C.A., Jackson, L., Tamplin, M.L., 1995 - Vibrio cholerae, V.
parahaemolyticus, V. vulnificus, and other Vibrio spp . In: Bacteriological
Analytical Manual, 8th ed., pp. 9.019.25. Washington, D.C. Food and Drug
Administration.
2. Gooch J., DePaola A., Kaysner C., Marshall D., 1996 - Evaluation of two direct
plating methods using nonradioactive probes for enumeration of V.
parahaemolyticus in oysters, Appl. Environ. Microbiol., 67 (2): 721-724.
3. ICMSF - International Commission on Microbiological Specifications for Food. Vibrio
parahaemolyticus, In: Roberts T.A. (chairman Ed.), Microorganisms in foods, 5th
edition: Characteristics of microbial pathogens, Blackie Academic Professional,
London, New York, Melbourne, Tokyo, 1996, 426-435.
4. McCarthy S.A. DePaola, A., Cook D.W., Kaysner C.A., Hill W.E., 1999 - Evaluation
of alkaline phosphatase- and digoxigenin-labelled probes for detection of the
thermolabile hemolysin (tlh) gene of Vibrio parahaemolyticus, Letters in Applied
Microbiology; 28(1): 66-68.
5. Nordstrom J.L., DePaola A., 2003 - Improved recovery of pathogenic Vibrio
parahaemolyticus from oysters using colony hybridization following enrichment,
Journal of Microbiological Methods, 52: 273-277.
6. Taniguchi, H., Hirano, H., Kubomura, S., Higashi, K., Mizuguchi, Y., 1986 -
Comparison of the nucleotide sequences of the genes for the thermostable direct
hemolysin and the thermolabile hemolysin from Vibrio parahaemolyticus.
Microbial Pathogenesis; 1: 425 432.

ACKNOWLEDGEMENTS

We would like to thank Prof. Daube Georges and Dr. China Bernard of the Liege
University, Belgium, for their assistance in design and development of primers and probes
for use in this work.

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REAL TIME PCR WITH SYBRGREEN TO DETECT


THE PATHOGENICALLY STRAINS OF
VIBRIO PARAHAEMOLYTICUS
Gh. BRDAN
U.S.A.M.V. Iai

Vibrio parahaemolyticus (Vp) is an estuarine bacterium that is the


principal cause of shellfish associated cases of bacterial gastroenteritis
worldwide. There are pathogenic only the stocks which have the factors of
virulence: the thermostable direct haemolysin (TDH) and TDH-related
haemolysin (TRH). The thermostable direct haemolysin gene (tdh) and the
thermostable-related haemolysin gene (trh) are both currently used as
markers of pathogenicity for this micro-organism. The paper presents an
alternative of the real time PCR, with SYBR Green, useful for the
simultaneous detection of these genes. The principle of the analysis is to
identify the melting curves, with the peaks of fluorescence at the specific Tm
for each amplicon.

Key words: real time PCR, V. parahaemolyticus, SYBRGreen

INTRODUCTION

The details of the pathogenesis of the V.p. are known since the years '50,
when a bond was established between virulence for the man of an isolate of this
bacterium and its capacity to produce a haemolyse on agar with blood
(Wagastuma agar) - phenomenon of Kanagawa (3). In Japan, it was observed that
96,5% of 2720 human stocks were positive (K+ stocks) compared only 1% of 650
environmental stocks (stocks K-). Other authors brought back frequencies even
weaker of K+ stocks among the stocks of the environment. There are
experimental arguments in favour of a natural selection of the K+ stocks in the
intestinal tract and a better survival of the K+ stocks in the ecosystems estuaries.
Currently, it is knows three haemolysins at V.p.: thermostable direct
haemolysin-TDH; thermostable related hemolysin-TRH; thermolabile haemolysin
-TLH. The food toxi-infection is caused by TDH and/or TRH but only if this one
is produced in the digestive tract (4). The stocks producing one of two
haemolysins TDH or TRH (or more rarely both) is represent only a little part of
the population of V.p. This organism is frequently isolated from coastal waters
and seafood in temperate regions throughout the world. It is the most frequent
cause of foodborne disease in Japan, where many residents eat raw fish. A
number of common-source gastroenteritis outbreaks attributed to V.p. have been
associated with oyster consumption (2).

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The main target of this experiment was to development of a multiplex real-


time PCR assay with SYBRGreen for the pathogenic (tdh+ and trh+) V.p.

MATERIALS AND METHODS

The tests were developed in the Laboratory of microbial genetics of Faculty


of Veterinary medicine of Liege (Belgium).
Genomic DNA preparation. Selected V.p. strains [(tdh+), (trh+), (tdh+,trh+),
and (tdh+,trh+)] used for real-time PCR bacterial detection from the Centre for
Environment, Fisheries and Aquaculture Science CEFAS (Weymoth, England).
Design of primers and fluorogenic probes for real-time PCR. The complete
nucleotide sequences for all reported variants of the tdh and trh genes of V.p.
were aligned and compared by GenBank to design oligonucleotide primers and
TaqMan fluorogenic probes targeting regions for each of these genes. Multiple
primer sets were designed and tested for specificity and compatibility in multiplex.
Information on the primers and probes used in the assay can be seen in the
following table:
Table 1
Primers and probes

Target ToC Amplicon ToC


Sequence Gene
primers hybrid. size (bp) melting
tdh 499U17 5-gct-ata-agc-gcg-gtc-at-3 tdh 50,7C 324 80C
tdh 803 L20 5-cat-tgg-cat-gca-taa-att-ag -3 tdh 50,7C 324 80C
trh 37 U24 5-ttg gct tcg ata ttt tca gta tct -3 trh 54,1C 486 83,3C
trh 498 L25 5- cat aac aaa cat atg ccc att tcg-3 54,1C 486 83,3C

Real-time PCR amplification. The real-time PCR cycling protocol,


fluorescent detection parameters, and reaction component concentrations were
carefully optimized for the simultaneous detection and quantification of the tdh,
and trh genes. PCR was conducted in a 25 l volume using the following reaction
components (final concentrations shown): 12,5 l SYBRGreen SuperMix with
ROX (BioRad Laboratories, Hercules, CA, USA, 2004) [2x tampons de reaction;
0,4 mM dATP; 0,4mM dCTP ; 0,4 mM dGTP; 0,8 mM dUTP; 50 U/ml enzyme
iTaq DNA polymerase; 6 mM Mg2+; SYBRGreen I dye; 1 M ROX internal
reference, stabilisations]; 400 nM of each of the dNTPs (Sygma Genosys), 250
nM of each primer (described above), and 9.75 l ultrapure water. Real-time PCR
thermal cycling was conducted using the ABI Prism 7000 (Applied Biosystems).
The negative control (sterile water added as template in the reaction) were
prepared for each PCR master mix. For each reaction, a plot of the cycle
threshold (Ct) vs. fluorescence (and log fluorescence) was examined for the FAM
channel of the ABI Prism 7000 instrument.
Cycling parameters. The two-step touchdown cycling parameters used are
shown below:

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Table 2
Cycling parameters

Etape No. cycles Temperature Time


1 1 Inactivation of uracyl-N-glycosilase 50C 2 min.
2 1 Denaturation/taq activation 95C 3 min.
Amplification Dnaturation 95C 15 sec.
DNA segment,
3 40
collect and Hybridation and
60C 60 sec.
analyse data) extension

The ABI Prism 7000 instrument was programmed to measure the


accumulated fluorescence in each reaction tube at the end of each amplification
cycle using the sequence detection system. A sample was therefore considered
to be positive when a signal of at least eight fluorescence units above baseline
(after background correction) was observed in the FAM channel of the instrument
within 40 cycles.

RESULTS AND DISCUSSIONS

To obtain distinct curves of dissociation one carried out several real time
PCR with different combinations from the primers: mix A (tdh), mix B (trh) and
mix C (tdh and trh). The dissociation curve of the sample tdh+ and trh+ had two
peaks, one, discrete, with 74C for tdh+ and another, very full, with 77C, for the
trh. Only for the mix with the tdh+/trh+ ratio of 10/1 had a curve of dissociation
with two peaks of comparable intensity (Fig. 1).

rh
dh

Fig. 1. Multiplex melting curves

When a stock of V.p. was highlighted on a selective agar, it is necessary to


push the analysis further. Indeed, all the stocks of V.p. are not pathogenic. It is
necessary to check the presence of genes of virulence tdh+ and trh+. The

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detection of Vp by the PCR in real time with system SYBR Green, is one of the
techniques of bacterial genetics least expensive. On the basis of work completed
by Japanese and American teams (1, 2), techniques of fast detection of the
bacterium by the PCR and the techniques with DNA probes, having as target the
gene tdh were developed. A recent study in which the SYBR Green I-based PCR
was compared with TaqMan assay for detection of faecal pathogens showed that
the two assays had the same level of sensitivity of detection (1).
Direct detection of the genes tdh+ and trh+ by the PCR in real time with
system SYBR Green were gravitational in the identification by the PCR in real
time, using specific TaqMan probes. The principle of the analysis is to identify
the curves of dissociation, with a peak of fluorescence at the temperature of
hybridization (Tm) specific for each amplicon (5). One used samples of DNA of
Vp., checked for two genes: a sample contained DNA with only the gene tdh+,
another sample, environmental, contained neither of tdh+ not of trh+, and the
third, pathogenic, was positive for the tdh and the trh.

CONCLUSIONS

SYBRGreen (SG) will preferentially bind to selected amplicons during


multiplex real-time PCR. With an increasing demand for high-throughput and
multiplexing assays, the characteristics of SG described here may reduce
optimisation times by avoiding the use of limiting concentrations of SG in assays.
Rapid detection of V. parahaemolyticus pathogen in consumable oysters
and in coastal water, especially in and around approved oyster-harvesting sites, at
a level that meets the recent guidelines would help to reduce the incidence of
illness and fatality that result from ingestion of raw shellfish or from exposure to
coastal water.

REFERENCES

1. Fukushima H, Tsunomori Y, Seki R., 2003 - Duplex real-time SYBR green PCR
assays for detection of 17 species of food- or waterborne pathogens in stools. J.
Clin. Microbiol; 41(11): 5134-5146.
2. Kaysner Ch., DePaola A.Jr., 2004 - Vibrio, In: Bacteriological Analytical Manual
Online, 8th Edition, Chapter 9, substantially rewritten and revised,
url://vm.cfsan.fda.gov/~ebam/bam-9.html.
3. Nishibuchi M, Kaper JB., 1995 - Thermostable direct hemolysin gene of Vibrio
parahaemolyticus: a virulence gene acquired by a marine bacterium. Infect.
Immun; 64:2093-2099.
4. Peterson K., 2000 - Molecular pathogenesis of Vibrio infections. In: Carry J.W., Linz
J.E. et Bharnagar D. (Eds.) Microbial Foodborne Diseases mechanisms of
pathogenesis and toxins synthesis, Technomic Publishnig Co. Inc., Lancaster,
USA, 157-183.
5. Robert-Pillot A., Guenole A., Lesne J., Delesmont R., Fournier J.M., Quilici M.L.,
2004 - Occurrence of the tdh and trh genes in Vibrio parahaemolyticus isolates
from waters and raw shelfish collected in two French coastal areas and from
seafood imported into France, Int. J. Food Microbiol.; 91: 319-325.

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OBSERVATIONS ON THE FREQUENCY OF


YERSINIA ENTEROCOLITICA IN PIGS
OBSERVAII PRIVIND FRECVENA YERSINIEI ENTEROCOLITICA LA SUINE

M. CONDREA, M. CARP-CRARE,
T. PERIANU, Elena VELESCU
U.S.A.M.V. Iai

230 pig samples were bacteriologically tasted during slaughters in


slaughter houses, aiming at the presence of Yersinia enterocolitica. The
samples were represented by the intestinal content and curettage of the
intestinal mucous membrane, drawings from the tongue and skeleton. The
method used was the one Enterobacteraceae, adapted to the biologic
peculiarities of Yersinia enterocolitica. 9 stems of Yersinia enterocolitica
were isolated, the isolation percentage being of 3.91%. The highest
isolation frequency was registered from the curettage of the intestinal
mucous membrane 6 stems (6.66%). Out of the 9 stems, 3 (5.17%) were
isolated in spring, 2 (3.5%) in autumn, 2 (3.5%) in winter and 1 (1.75%) in
summer.
The isolated stems belonged to 2 biotypes (1 and 4), biotype 4 (7
stems 77.7%) being predominant. Biotype 1 comprised 2 stems (22.3%).
The serotipia shows the predominance of the O3 serotype (7 stems),
a serotype which was also frequently isolated from the clinical and
asymptomatic forms in people (7, 8, 10). Two stems, comprised in biotype
1, belonged to serotypes O4 and O5. We notice that besides the serotypes
traditionally considered as responsible for this disease in people O3, also
occur new serotypes like O4 and O5 frequently isolated from the
environment and considered till very recently as being neopathogenic. The
frequency and variety of serotypes and biotypes isolated both from animals,
especially the pig, and from people in clinical and asymptomatic cases,
draw the attention on new epidemiologic situations.

Key words: Yersinia enterocolitica-pig-frequency

Many zoonoses have spread during the past few years to different regions
of the world and their frequency index has increased, thus endangering public
health by the variety of contamination possibilities. Yersiniosis with Yersinia
enterocolitica is among them.
The implications of this bacterium in the food chain in people started an
increasing interest in the last few years among researchers from the whole world
(1, 4, 7, 8). This bacterium with a high bacteriologic and epidemiologic
heterogeneity (4, 7, 8) was isolated from an important number of domestic and

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animals and from the environment (1, 2, 3, 9, 10). It is capable of disturbing


public health both by its virulence and by its toxigenic capacity (6).
Having regard to the great plasticity of the pathogenic agent and the
possibility of adaptation to new species it is highly necessary to permanently
discover this species, to evaluate its incidence in pigs, to evaluate the frequency
and circulation of serotypes and biotypes known as with implications in human
pathology, as well as serotypes which in the last few years modified their ecologic
determinism and their pathogenic capacity.

MATERIAL AND METHODS

230 pig samples taken during slaughtering were bacteriologically


investigated. Each sample was represented by intestinal content and curettage of
the intestinal mucous membrane, drawings from the tongue and skeleton.
The methodology used was the one for enterobacteriaceae, adapted to the
biologic peculiarities of Yersinia enterocolitica (4, 7).

RESULTS AND DISCUSSIONS

The bacteriologic investigation of the 230 samples allowed the isolation of


9 stems of Yersinia enterocolitica; the isolation percentage was of 3.91%.
This index of 3.91% places pigs among the main sources of infection with
Yersinia enterocolitica. Back in 1971, Mollaret H.H. (4) issued the assumption
according to which the pig would be the main source of infection for people and
suggested the orientation of the investigations towards pigs. Ulterior research (2,
4, 9) confirmed this assumption by demonstrating the identity of the pig origin
cultures with the ones isolated from the clinical cases in people.
The results are comparable to the ones reported in the Scandinavian
countries, Belgium and Netherlands, where the isolation percentage was of 3-5%
(Acha, 1989). Both in Romania and in Belgium, Sweden, Denmark, Nigeria,
Canada the incidence of Yersinia enterocolitica in sick people with acute diarrhea
syndrome is between 1-4%, showing thus a slight connection between the pig
carrier and the cases of diarrhea with this bacterium (2, 6, 7, 8).
The stems were isolated only from the curettage of the intestinal mucous
membrane, 6 (66.6%), from curettage and drawings from the tongue 2 (22.2%),
from curettage, drawings from the tongue and skeleton 1 (11.1%). In Italy,
Mazzeo (1993) isolated Yersinia enterocolitica 2.8% from the feces and 5.6%
from the tonsils. Although some authors report increased frequencies of Yersinia
enterocolitica in tonsils and on the pigs tongue, feces still represent the most
frequent and constant source, which is also similar with our research.
The 230 samples were equally distributed for the 4 seasons, that is spring,
summer, autumn and winter. Out of 9 stems, 3 (5.17%) were isolated in spring, 3
(5.17%) in autumn, 2 (3.5%) in winter and 1 (1.75%) in summer. The higher

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frequency of these isolations in the cold season is the feature which makes this
bacterium different from the enteric infections which evolve with a high
frequency in summer. The pick autumn winter spring in Romania (4, 10) is
similar with other countries reports, October January: Belgium, November:
Sweden, September November: Finland, December March: Chech Republic
(8).
The isolated stems belonged to the 2 biotypes (1 and 4), but biotype 4 (7
stems 77.7%) was predominant. Biotype 1 comprised only 2 stems (22.3%).
The serotipia shows the predominance of serotype O3 (7 stems), a serotype
frequently isolated also from the clinical and asymptomatic forms at people (8,
10). Two stems, the ones comprised in biotype 1, belonged to serotypes O4 and
O5.
Although the serotypes traditionally considered responsible for the human
disease (4, 8, 10) were O3 and O9 and less O5, 27, during the last few years there
is an increasing variety of serotypes depending on the disease; the variety is also
in the case of the pig carrying this type of bacterium. Thus the presence in pigs of
the serotypes O4 and O5 (biotype 1) which so far were considered wild non-
pathogenic demonstrates the possibility for this bacterium to adapt under the
influence of biologic pressures and announces a new epidemiologic situation.
The same variety of the isolated serotypes, both from animals and from the
human clinical cases, is more and more relevant. Chiesa and his collaborators
(quoted by Felip, 1989) isolated many serotypes from the patients with diarrhea in
Italy: O3 (57%), O6 (6.5%), O8 (6%), O9 (5%). The same author isolated mainly
from clinically healthy pigs the same serotypes: O3 (29.2%), O6 (10.3%), O9
(9.1%), O10 (9.7%), O7,8 (10.9%). The conclusion is the predominance of
biotypes 4 and 1.

CONCLUSIONS

1. 9 stems of Yersinia enterocolitica were isolated from pigs; the isolation


percentage was of 3.91%.
2. The 9 stems of Yersinia enterocolitica belonged to biotypes 4 (7 stems)
and 1 (2 stems) and serotypes O3 (7 stems), O4 (1 stem) and O5 (1 stem).
3. In the cold seasons spring, autumn, winter the isolation frequency
was higher (8 stems), as compared to the warm season summer (1 stem).
4. The highest frequency of isolations was from the intestinal content (6
stems).

REFERENCES

1. Acha N.P., Szyfres B., 1989 - Yersiniose enterocolitique - Zoonoses et maladies


transmissibles communes l'homme et aux animaux. Deuxime dition, 207-212

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2. Adesyun A.A., Kaminijolo J.S., Loregnard R., 1993 - Frequency of Isolation


Yersinia enterocolitica from Livestock in Trinidad, Vet
3. Carp Carare M, Timofte D, Condrea M, Guguianu E, Carp Carare C, Solcan G,
2001 - Study of ecology and epidemilogical implications of Campylobacter and
Yersinia bacteria. Buletinul Universitatii de Stiinte Agricole si Medicina
VeterinaraCluj-Napoca, Seria Medicina Veterinara, 55-56,
4. Condrea M., 1996 - Cercetari bacteriologice si serologice asupra Yersiniei
enterocolitica izolata de la animale, Teza de doctorat, UAMV
5. Condrea M., Straton A., 1994 Frecventa Yersiniilor la animale domestice si unele
elemente de mediu, Lucr. st. zoot., Iasi 19-20
6. Cornelis G.R., 1995 - Le Plasmide pYV lment cl de la virulence de Yersinia, M.
C. 11, 9, 1295-1304
7. Cover T.L., Aber R.C., 1989 - Yersinia enterocolitica, New Engle. J. Med., 321, 1,
16-24
8. Felip G.D., Orefice L., Medici D., Germoni S., 1989 - Raspndirea, patogeneza si
prevenirea Yersiniozei cu Yersinia enterocolitica, Slez. vet. 30, 1, 15-18
9. Mazzeo A., Petrarca G., Iannito G., Calavita G., Porta L., 1993 - Yersinia
enterocolitica: Isolation from Faeces and Tonsils of Pigs Slaughtered Normaly.
Vet. Bul. 63, 5, 458.
10. Straton A., Staton C., Beldiman N., Condrea M., 1992 - Incidenta Yersiniei
enterocolitica la om si la unele specii de rozatoare ca posibila sursa de infectie.
Congr. Nat. de biol. II, Iasi 196 197 V, Ed. Acad.

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SOME ASPECTS REGARDING THE METHODOLOGY


OF ISOLATING YERSINIA ENTEROCOLITICA
UNELE ASPECTE PRIVIND METODOLOGIA DE IZOLARE A
YERSINIEI ENTEROCOLITICA

M. CONDREA, M. CARP-CRARE,
Eleonora GUGUIANU, C. CARP-CRARE
U.S.A.M.V. Iai

60 stems of Yersinia enterocolitica were bacteriologically searched,


following some peculiarities regarding their morphology, mobility,
breeding conditions and cultural aspect in order to improve the isolation
methods in the veterinary labs.
We rendered evident the pleiomorphism characteristic to the
Enterobacteriaceae, mobility to 29oC and immobility to 37oC; direct
insemination, without enrichment, adapts better in the veterinary labs; the
slower development on the media for Enterobacteriaceae makes it
necessary the repication 48 hours later, when the colonies become visible.

Key-words: Yersinia enterocolitica-isolating

As part of the zoonoses group, Yersinia enterocolitica is the cause of the


acute diarrhea in people, having as a possible source of infection some animals
(infected or carriers) and an indirect way of transmission through contaminated
food and water (1, 5, 7). Although the bacterium was found in a big number of
animals from different species, proving the identity of the isolated stems with the
ones that started the disease in people (7), there is no proof of a direct
transmission. Besides its epizoothologic role (source of infection), in the last few
years researchers quote more and more the implication of Yersinia enterocolitica
in veterinary pathology (5, 7).
Now we have at our disposal a variety of data regarding the biology of
Yersinia enterocolitica. Nevertheless there are still many unknown things,
generated by the biologic peculiarities of this bacterium. Yersinia enterocolitica is
approached more at the level of the research labs, without being the subject of
current investigations in diagnosis labs. The every day more frequent implication
in human and veterinary pathology makes it necessary the introduction of this
bacterium in the routine investigations of the speciality labs.

MATERIAL AND METHODS

60 stems of Yersinia enterocolitica were bacteriologically investigated after


being isolated from pigs (44 stems), taurines (5 stems), and dogs (11 stems). We

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aimed at its morphology, mobility, breeding conditions and cultural aspect on the
media used for the isolation of Enterobacteriaceae, following the biologic
peculiarities of this species in order to improve the isolation methods.

RESULTS AND DISCUSSIONS

In all the speciality works Yersinia enterocolitica is presented as a Gram


negative, non-sporulated, non-capsulated bacterium (1, 5, 7, 8, 11). The classical
form is of coccus-bacillus (1, 11) with the tendency of pleiomorphism according
to the incubation temperature, age of culture, mediums composition, and biotype.
The pleiomorphism of Yersinia enterocolitica, mentioned in the speciality works
(1, 7, 11) is a feature of the Enterobacteriaceae.
In our research on the smear made of cultures in liquid media (peptonated
water, simple tomato paste) there is a predominance of the oval coccus forms,
when incubation was made at 22oC for 24 hours, while incubation at 37oC leads to
the occurrence of bacillus forms.
In the cultures on the solid media there is a predominance of short and thick
bacilli. In old cultures pleiomorphism is very much stressed.
Having regard to the studied smears we may confirm that the bacterium
does not have a characteristic way of grouping; the singular forms coexist with
the short chains and piles. The tendency towards bipolar colouring is present only
on the smears made from cultures on liquid media.
The results regarding the morphology of the isolated stems are compatible
with the existing data (1, 7, 13).
The fundamental feature of Yersinia enterocolitica referring to the
synthesis of the cilia according to the incubation temperature is also present in the
60 studied stems. The best temperature allowed to render evident the mobility is
between 22-30oC (1, 2, 4, 11).
In our research of the 60 stems we indirectly studied the presence of the
cilia, following the mobility on the medium M.I.U. (mobility indole urea) at
29oC and at 37oC. All the stems were mobile at 29oC and immobile at 37oC.
Nevertheless 5 of the studied stems were lately slightly positive at 37oC. These
stems of pig and canine origin were comprised in biotype 1 (O4, O5, O6) and
biotype 2 (O8).
Out of the 7000 stems from the National Centre Collection in Paris,
investigated by Bercovier and his collaborators, 1980 (1) regarding the mobility at
28oC, 88% were mobile and 10% had a late reaction (mobile 3-4 days later).
The conclusion was that the intensity of this phenomenon is slightly
variable depending on the stem.
The mobility of Yersinia enterocolitica at a lower temperature than 30oC
and the lack of mobility over this temperature is a feature of all the species of the
gender Yersinia, except for Yersinia pestis which is immobile.

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Analysing the results regarding the mobility of Yersinia enterocolitica we


conclude that, if for the rest of the species mobility is a helping test, for the
identification of Yersinia enterocolitica it is an essential test which is
recommended to be the first one when we suspect that we have to do with
Yersinia according to the cultural and morphological features.
All the stems isolated grew better at a pH of the media between 6.8-7.6.
The researches (7, 11) have pH limits 5-9 and as the best pH: 7.2-7.4.
According to Bougeois, 1990 (2), the minimum pH is 4 and the maximum one is
10. The bacterium may survive for 48 hours both at a pH of 3.6 (Kendal and
Gilbert, 1980) and at a pH of 10-12 in alkaline conditions. Doyle (6)
recommended the treatment with potassium 0.25-0.5% for the cultures after
enrichment, aiming at inhibiting the other enterobacteriaceae, much more
sensitive at the alkaline shock than Yersinia.
The temperature area in which Yersinia enterocolitica develops is
comprised between large limits, 4-43oC (11). The researches (1, 11) show as
minimum temperature for development 4oC a temperature at which micro-
organisms multiply, but the rhythm of development is slow. As Mehlman (10)
states, the development of Yersinia enterocolitica at a temperature of 4oC is due to
inhibition of the association flora, consequently to the reduction of the
interference of the metabolits produced by other Gram negative germs.
Using in our researches as a method of enrichment peptonated water and a
4oC temperature with transfers to isolation media 7 or 14 days later, we found out
a slight increase of the isolation percentage. Out of the 60 stems, two (3.3%) were
isolated only on the variant with enrichment 14 days later, the rest of 58 being
isolated on both variants.
Besides the slight increase of the isolation percentage, enrichment in cold
conditions, 14 days, offers better conditions of repication for Yersninia
enterocolitica, by means of increasing the number of colonies on Petri plate and
an inhibition of the association flora. We consider the feature of Yersinia
enterocolitica of developing at temperatures of 0-4oC, without nutritive
exigencies, as an adaptation and not as an accident in its evolution, important to
the sanitary-veterinary control of food. Thus, cooling of food slows down the
multiplication of this bacterium, but it does not inhibit it completely (it visibly
increases in 2-5 days on meat extract at 4oC) (11).
Swaninathan (13) states that at 4oC the bacterium is better cultivated at a
neutral pH, but very few at pH 5.2-5.4.
Enrichment in cold conditions, 4oC in peptonated water, phosphate tampon
or tomato paste, uses the psychophril feature of the germ (Schieman).
Analysing the results, we may conclude that this bacterium, though it
develops at a cooling temperature, cannot be considered a psychophril because the
best conditions for its development are 22-25oC and even 37oC.

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Enrichment in the presence of selective sublayers often appeals to using the


Rappaport medium, sometimes additionally with carbenicilin or on the gall
oxalate sorbosis medium, with an incubation at 22-25oC, 1-5 days. According to
Schieman (12), this method has only the advantage of being fast as compared to
enrichment in cold conditions.
The medium of selenite novobiocine allows a better enrichment at 37oC,
but it does not offer selectivity (10).
Although there is a multitude of researches concerning the enrichment
conditions for Yersinia enterocolitica the conclusion is that there are not enough
factors which stimulate the development of Yersinia enterocolitica on the one
hand and factors which inhibit the development of the association flora on the
other hand. The only selective agent is the 4oC temperature.
No method seems so far usable in order to isolate or enrich the pathogenic
cultures of Yersinia enterocolitica and disadvantage the non-pathogenic ones or
the other species of Yersinia, except maybe the modified Rappaport medium (1,
11) which selects the cultures O3 and O9, but it inhibits O8 and O5, 27.
Some researchers (7, 11) recommend different enrichment media according
to the serotype, pathogenic cultures etc.
Having regard to the results obtained by other researchers, as well as the
results of our researches regarding the variety characterizing this species, we
consider that we should use some enrichment media to favour a wide isolation of
Yersinia serotypes under the conditions in which the dynamics of existence
imposes an increase of variety.
The best development for Yersinia enterocolitica is a temperature between
22-29oC (2, 11). Zarnea (1994) states that we cannot entirely talk about the best
temperature for a certain species, but about the best temperature for each of its
biologic activities.
We noticed at the stems that we studied a faster disturbance of the tomato
paste at 37oC, as compared to the incubation at 22oC. Nilehn and Wauters (quoted
by Rusu) demonstrated that the highest number of cells is accumulated at 25oC,
when the exponential stage is the longest, while at 34oC, when division is very
fast, the quantity of biomass is insignificant.
The stems of Yersinia enterocolitica studied develop well on the media for
Enterobacteriaceae, without adding breeding factors (peptonated water, simple
tomato paste, Mac Conkey, Leifson, Drigalski, simple gelose).
The variant of isolation through direct insemination on Leifson medium
and partially on Mac Conkey offered an index of positivism of 96.7%. Both
Leifson and Mac Conkey medium allowed the isolation of 58 stems, 2 less, as
compared to the enrichment variant.
Comparing the results obtained to the ones of other researchers (4, 8, 11),
the conclusion is that when it comes to the isolation of Yersinia enterocolitica
from the feces there is almost an unanimous agreement on the efficiency of the

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media based on gall salts and especially the ones with Leifson sodium
deoxycolate or its variants.
We aimed at the cultural aspect of Yersinia enterocolitica stems on liquid
and solid media according to the incubation temperature and the age of the
culture.
On the liquid media, cultures brought up disturbances at different speeds
depending on the incubation temperature. Old cultures have a deposit, sometimes
a membrane. It does not have pigments.
On the solid media the colonies of Yersinia enterocolitica are dull or
translucent, with regulate, non-pigmented margins. On the blood gelosis (sheep
blood defibrinated in proportion of 2%) it develops non-haemolytic colonies.
24 hours after incubation at 29oC or 37oC, the colonies of Yersinia
enterocolitica have dimensions between 0.5-1mm in diameter, at the limit of
visibility, contrasting with the other Enterobacteriaceae with dimensions between
2-4mm in diameter; the former ones may be repicated for identification.
In order to be repicated the incubation time of the cultures was prolonged to
48 hours.
This fact, also described in the speciality works (1, 5, 11) is useful within
the work methodology for the diagnosis labs. Due to the slow breeding of
Yersinia enterocolitica the inseminated plates for the isolation of
Enterobacteriaceae must be monitored after 48-hour incubation or left for one
day at the lab temperature and then checked again.

CONCLUSIONS

1. 60 stems of Yersinia enterocolitica were bacteriologically tested; the


analysis allowed us to select some biologic peculiarities of this bacterium in order
to isolate it in the veterinary labs using fast methods.
2. The stems of Yersinia enterocolitica studied came from pigs (44 stems),
taurines (5 stems) and dogs (11 stems).
3. The examination of the Gram coloured smear shows the polymorphism
characteristic to Enterobacteriaceae.
4. The mobility of the stems of Yersinia enterocolitica at 29oC and their
immobility at 37oC makes us recommend the introduction of this test when it
comes about drawing the differences between this and the other
Enterobacteriaceae, which are immobile at 29oC.
5. By studying isolation on the two variants (cold enrichment and direct
insemination) we found out that the direct method adapts better to the
methodology of work in the diagnosis labs when it comes to the isolation of
Yersinia enterocolitica from feces.
6. On the solid media, Yersinia enterocolitica develops slower as compared
to the other; this peculiarity makes Yersinia get away from repication once with

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the other bacteria. While on the Leifson medium, after 24-hour incubation at
37oC, the colonies are under the limit of visibility, a prolongation of incubation up
to 48 hours being necessary for the isolation, on the Mac Conkey medium the
development is faster, the repication being possible 24 hours after the incubation,
once with the other Enterobacteriaceae.

REFERENCES

1. Bercovier H., Brenner D. J., Ursing J., Steigerwalt A. G., Fanning G. R., Alonso
J.M., Carter G.P., Mollaret H.H., 1980 - Characterization of Yersinia
enterocolitica sensu stricto, Current Microb., 4, 201-206
2. Bourgeois C.M., Mescle J.F., Zucca J., 1990 - Yersinia enterocolitica, Microb. alim.
Tom. 1 Colect. Scien. the. agro-alim., Paris 117-124
3. Carp-Carare M., Timofte D., Condrea M., Guguianu E., Carp-Carare C., Solcan
G., 2001 - Study of ecology and epidemilogical implications of Campylobacter
and Yersinia bacteria. Buletinul Universitatii de Stiinte Agricole si Medicina
Veterinara Cluj-Napoca, Seria Medicina Veterinara, 55-56,
4. Condrea M., 1996 - Cercetari bacteriologice si serologice asupra Yersiniei
enterocolitica izolata de la animale, Teza de doctorat, UAMV
5. Cover T.L., Aber R.C., 1989 - Yersinia enterocolitica, New Engle. J. Med., 321, 1,
16-24
6. Doyle M.P., Hugdahl M.B., 1983 - Improved Procedise for Recovery yersinia
enterocolitica from Meats, Appl. and Environ. Microb., 45, 1, 127-135
7. Felip G.D., Orefice L., Medici D., Germoni S., 1989 - Raspandirea, patogeneza si
prevenirea Yersiniozei cu Yersinia enterocolitica, Slez. vet. 30, 1, 15-18
8. Jacober F.X., 1968 - L'infection Yersinia enterocolitica. Une zoonose d'avenir,
Thse doctorat vtrinaire, Paris
9. Kendal M., Gilbert M.J., 1980 - Survival and Growth of Yersinia enterocolitica in Broth
Media and in Food. Microb. Grow. and surv. Extrem. of Env., Ed. Gould. 215 - 226.
10. Mehlman I.J., Ausilio C.G., Sanders A.C., 1978 - Problems in the Recovery and
Identification of Yersinia from Food. J. Assoc. off. Anal. Chem., 61, 4, 761 - 771.
11. Rusu V., 1985 - Bacteriologie medical vol. II (Blbie V. i col.), Ed. Medical, Bucureti,
345, 356 304-312
12. Shieman D.A., 1983 - Comparaison of Enrichment for Recovery of Virulent Strains of
Yersinia enterocolitica from Inoculated Beef Stew. J. F. P. 46, 11, 957-964
13. Swaminathan B., Harmon M.C., Mehlman I.J., 1982 - A Review: Yersinia
enterocolitica J. Appl. Bact. 52, 151 - 183.
14. Straton A., Staton C., Beldiman N., Condrea M., 1992 - Incidenta Yersiniei la om
si la unele specii de rozatoare ca posibila sursa de infectie. Congr. Nat. de biol.
II, Iasi 196 197 V, Ed. Acad.
15. Zarnea G., 1994 - Tratat de microbiologie general V, ed. Acad.

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ASPECTE CLINICE N GRIPA AVIAR


LA LEBDA DE VAR
CLINICAL ASPECTS OF THE AVIAN FLU AT SUMMER SWAN (CYGNUS OLOR)

M. ARSENE, Marinela ARSENE


D.S.V.S.A. Tulcea
Gh. SAVUA, T. PERIANU
U.S.A.M.V. Iai

The research was made in the Delta of Danube piscicultural


complex, Maliuc, district Tulcea.During 07-14.10.2005 were examined the
birds of passage wich procured their meal from the basins belonging to the
piscicultural complex.
After our research we found out decay and decease at the summer
swan, especially at the young birds.Tthe first signs noticed were the nervous
signs.

Key words: avian influenza, summer swan, clinical diagnosis

Influena aviar, cunoscut i sub numele de gripa aviar sau pesta aviar,
este o boal infecioas , contagioas care afecteaz mai multe specii de psri
domestice i slbatice. (4).
Agentul etiologic al influenei aviare este un virus ce face parte din genul
Influenzavirus A, familia Orthomyxoviridae. (2, 4).
Sursele de infecie sunt reprezentate de psrile bolnave sau aflate n
perioada de incubaie, de cadavrele psrilor infectate, de resturile de la
prelucrarea carcaselor i apele de la splarea acestora, de secreiile i excreiile
psrilor infectate ca i de produsele i ustensilele ce au venit n contact cu virusul
n diverse circumstane. (1).
Populaiile de psri slbatice, ndeosebi cele acvatice, reprezint un
important rezervor , care ntreine i vehiculeaz cele mai diverse subtipuri de
virus gripal. (1).
Virusul patrunde n organism pe cale respiratorie i mai puin pe cale
digestiv. Perioada de incubaie este obinuit de 3-7 zile, fiind dependent de
patogenitatea virusului. Clinic, boala evolueaz de regul supraacut , cu moartea
subit a psrilor afectate sau n decurs de 24 - 48 ore , n unele cazuri
manifestrile putnd mbraca forme acute, cu o durat de cel mult o sptman.
Manifestrile clinice exprimate sunt urmarea tulburrilor circulatorii,
respiratorii, digestive i ale sistemului nervos. Ele pot fi foarte diferite i
dependente de mai muli factori, inclusiv de vrsta i specia psrilor afectate, de
sistemul de exploatare i desigur de patogenitatea tulpinii de virus. (1, 2, 4).

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MATERIAL I METODE

Investigaiile au fost efectuate la psrile slbatice din Delta Dunrii i n


special la un crd de lebede de var (Cygnus olor), localizat pe amenajarea
piscicol din vecintatea localitilor Maliuc i Vulturu, n luna octombrie 2005,
cnd a fost declarat i primul focar de influen aviar la psrile slbatice din
Romnia. Ferma piscicol respectiv aparine societii comerciale S.C. Maliuc
S.A. i are o suprafa de aproximativ 2000 hectare , fiind format din mai multe
canale i 46 bazine. Colonia de lebede investigat era constituit att din psri
juvenile ct i adulte. S-au efectuat observaii clinice i s-au recoltat cadavre i
probe de snge precum i tampoane cloacale pentru examene de laborator.

REZULTATE I DISCUII

Ca urmare a observaiilor efectuate n perioada 7.10- 14.10.2005 la


amenajarea piscicol Maliuc , s-au constatat mortaliti la psrile slbatice din
acea zon i n special la lebedele de var (Cygnus olor).
n data de 07.10.2005, n aceast zon s-a observat prezena unei
aglomerri masive de lebede precum i existena unui numr de 45 lebede
moarte, cadavrele fiind n diferite stadii de alterare.
Aglomerarea de lebede a fost estimat la aproximativ 500 de exemplare,
format din tineret i din psri adulte. n urma evalurii fcute s-au identificat
139 psri moarte i 98 bolnave. Din cele 237 de cazuri recenzate, proporia cea
mai mare o reprezentau psrile tinere (70% ).
Datorit anotimpului rece de iarn, cnd cantitatea de hran este mai mic,
lebedele de var au rmas la iernat pe teritoriul Deltei Dunrii i s-au adunat n
grupuri mari n bazinele piscicole , acolo unde apa era mic i se mai putea gsi
hran. Densitatea crescut a psrilor a jucat un rol important n difuzarea bolii.
Exemplarele bolnave dintr-un crd de lebede de var puteau fi vzute uor,
deoarece acestea aveau un comportament atipic unei psri sntoase i anume:
nu se mai hraneau, rmneau n urma celorlalte lebede, nu i mai puteau lua
zborul sau dac reueau s se ridice de pe luciul apei zburau foarte puin (Fig. 1),
dnd semne de oboseal i de lips a coordonrii la aterizare. Psrile erau
epuizate iar pe luciul apei stteau abtute, n btaia vntului care le mpingea ctre
maluri, unde puteau fi prinse uor cu mna (Fig. 2).
Principalele manifestri clinice observate la psrile bolnave au fost
semnele nervoase. Dupa o anumit perioada de acalmie, pe care o prezentau
psrile n agonie, urmau crize nervoase de 30 secunde, caracterizate prin
tremurturi ale capului i gatului , prin contracii spasmodice ale membrelor,
ataxie, torticolis.

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Fig. 1. Lebede n ncercarea de a-i lua zborul.

Fig. 2. Capturarea lebedelor cu mna liber.

Pe lng semnele nervoase, un numr redus de psri prezentau i semne


respiratorii, cum ar fi secreii perinazale (Fig. 3), dispnee i respiraie zgomotoas.
La un numr nesemnificativ de cazuri s-a observat i forma digestiv,
caracterizat prin diaree apoas, de culoare galben-verzuie.

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Dac n prima zi de observaie psrile bolnave erau n agonie, cu diverse


manifestri clinice, a doua zi aceste psri erau gsite moarte (Fig .4) la malul
apei, orientate n funcie de direcia vntului.
S-au mai observat i alte semne clinice cum ar fi: mucoasa conjuctival
congestionat i edemaiat, depresie puternic asociat cu horiplumaie , moarte
subit fr manifestri clinice.
Au fost afectate n numr mai mare psrile juvenile i n numr mai mic
adultele.

Fig. 3. Scurgeri nazale la lebd cu influen.

Fig. 4. Cadavru de lebd ce plutete pe ap.

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Pe tot parcursul acestor zile s-au recoltat probe de la cadavre iar de la


psrile n agonie probe de snge (Fig. 5) , tampoane traheale i tampoane
cloacale (Fig. 6), care au fost trimise la Laboratorul Sanitar Veterinar Tulcea cu
suspiciunea de grip aviar A150. Psrile moarte au fost incinerate i ngropate
zilnic sub protecie de dezinfectant.

Fig. 5. - Recoltarea sngelui la lebd cu influen

Fig. 6. - Recoltarea tampoanelor cloacale la lebd.

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n data de 14.10.2005, Institutul de Diagnostic i Sntate Animal


Bucureti a confirmat suspiciunea de grip aviar i faptul c n zona Maliuc
Vulturu s-a constatat prezena virusului gripei aviare, tulpina H5N1, la lebedele de
var. Acestea coabitau cu alte specii de psri slbatice migratoare , cum ar fi
garlia, gsca mare, raa mic, lisia, pelicanul, tiganuul etc. care au jucat un rol
important n mbolnvirea lebedelor de var din aceast zon.

CONCLUZII

La lebedele din focarul de influen aviar examinat la Maliuc, boala a avut


o evoluie rapid. Cele mai afectate au fost psrile juvenile. Acestea nu i puteau
lua zborul i puteau fi prinse cu mna. La aterizare, prezentau tulburri de
coordonare i erau epuizate.
Principalele sindroame clinice au fost cele nervoase, urmate de semnele
respiratorii i n numr redus de semnele digestive.
Manifestrile nervoase au constat n crize de 30 secunde, caracterizate prin
tremurturi ale capului i gtului, contracii spasmodice ale membrelor, ataxie,
torticolis.
Semnele respiratorii s-au tradus prin secreii perinazale, dispnee i
respiraie zgomotoas.
S-a mai constatat depresie puternic, asociat cu horiplumaie i adesea
moarte subit, fr manifestri clinice.

BIBLIOGRAFIE

1. Brougere-Picoux Jeanne, 2006 - Le point sur linfluenza aviaire. Supplement


technique, nr. 97 a La Depeche Veterinaire.
2. Mnzat Moga R., 2001 - Boli infecioase ale animalelor. Ed. Brumar, Timioara.
3. Olaru E., 2005 - Instruciuni pentru recunoasterea Influenei aviare i diferenierea de
alte afeciuni ale psrilor. IDSA Bucureti.
4. Perianu T., 2005 - Boli infecioase ale animalelor. Viroze II. Editura Universitas XXI,
Iai
5. Savua Gh., 2001 - Epidemiologie veterinar general. Ed. Ion Ionescu de la Brad
Iai.
6. *** Programul aciunilor de supraveghere, prevenire i control al bolilor la animale,
ANSVSA Bucureti, 2005.
7. *** O.I.E., Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, 5th
edition,2004, www.oie.int.

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ASPECTE CLINICE N GRIPA AVIAR LA GINI


CLINICAL ASPECTS OF THE AVIAN FLU AT THE CHICKENS

M. ARSENE, Marinela ARSENE


D.S.V.S.A. Tulcea
T. PERIANU, Gh. SAVUA
U.S.A.M.V. Iai

The research was made in a farm stead in Vulturu, district of Tulcea,


placed near by the piscicultural complex where were find out deceases at
the summer swan. During 08-14.10.2005 were recorded deceases and
decays at the adult chickens. the domestic birds lived together with the
savages ones because of the bird- keeping system in the open. After our
research we find out the presence of the aviar flu virus at the domestical
birds who showed as principals signs: head and neck edema,crest cyanosis,
anorexia.

Key words: avian influenza, clinical diagnosis

Influena aviar (IA) sau pesta aviar clasic este o boal infecioas i
extrem de contagioas, ce se caracterizeaz prin tulburri generale grave, prin
simptome respiratorii, digestive i nervoase , nsoite de o infiltraie edematoas a
esutului conjunctiv i cutanat din regiunea capului i gtului. (2, 4).
Agentul etiologic este ncadrat n familia Orthomyxoviridae, genul
Influenzavirus, tipul A, suele nalt patogene aparinnd sub-tipurilor H5 i H7.
(1, 7). Infeciile cu virus nalt patogen, reduse ca inciden nc, nu trebuie
confundate cu cele cu virus slab patogen.
Influena aviar afecteaz n condiii naturale toate galinaceele, dar sunt
susceptibile i multe alte specii de psri domestice i salbatice (anseriforme,
passeriforme etc.). (1).
Ginile adulte sunt mai sensibile dect puii iar boala are un pronunat
caracter septicemic, cu evoluii supraacute i acute. Dup o incubaie de 3-5 zile,
manifestrile apar brusc, cu un aspect exploziv. (1, 2, 4, 7).
Semnele clinice mai importante sunt reprezentate de o stare depresiv
sever, inapeten, reducerea produciei de ou, edeme cefalice, tulburri
circulatorii, digestive, respiratorii, reproductive i ale sistemului nervos,
mortalitatea putnd atinge 100% din efectiv. (1, 3, 4).

MATERIAL I METODE

Studiul s-a efectuat la psrile domestice din gospodriile individuale ale


populaiei din localitile Maliuc i Vulturu din judeul Tulcea , localiti situate n

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Delta Dunrii. Cercetrile au fost fcute n luna octombrie 2005, cnd s-a
identificat i declarat focarul de grip aviar n aceste localiti. Primele semne
au aprut n gospodriile din vecintatea amenajrilor piscicole unde s-au
semnalat n aceeai perioad mbolnviri i mortalitate la lebedele de var.
Suspicionarea s-a bazat pe criterii epidemiologice i clinice iar confirmarea s-a
realizat prin examene de laborator (virusologic, PCR) efectuate la IDSA
Bucureti.

REZULTATE I DISCUII

Observaiile au fost fcute ntr-o gospodrie situat pe malul stng al


braului Sulina, n localitatea Vulturu din judeul Tulcea, n perioada 08 -
14.10.2005, cnd n aceast zon a fost declarat i primul focar de grip aviar la
psrile slbatice (lebda de var).
Ca urmare a suspiciunilor de grip aviar la lebedele de var (Cygnus olor)
din amenajarea piscicol care se afla n vecintatea gospodriilor populaiei, s-a
demarat aciunea de supraveghere a efectivelor de psri domestice susceptibile
de a contracta boala (A 150, dupa lista OIE), conform planului naional privind
supravegherea i controlul bolilor la animale (6).
n timpul supravegherii clinice, dup o metodologie bine stabilit (3), n
una din gospodriile din satul Vulturu s-au depistat dou cadavre de gin. Din
anamnez a reieit c n aceeai gospodrie, n cursul aceleeai zile, au mai murit
7 gini i dou curci, toate fiind psri adulte. n gospodrie mai existau 46 de
gini, din care 6 pui i 40 adulte , precum i doi pui de curc.
Deoarece efectivul de psri din gospodrie era crescut n sistem extensiv,
avnd acces liber la balt , se presupune c aceste psri au coabitat cu psrile
migratoare din amenajarea piscicol din vecintate .
S-au constatat mortaliti la psrile domestice ncepnd din data de
08.10.2005 (Fig. 1). n dimineaa urmtoare au mai fost gsite nc dou cadavre
de gini adulte i au fost observate alte cteva psri cu semne clinice.
n sintez, semnele clinice observate la psrile bolnave din focarul studiat
au fost: moarte subit fara manifestri clinice; adinamie i depresie puternic,
asociat cu horiplumaie; inapeten i sete excesiv; cianoza crestei i brbielor
(Fig. 2, 3); respiraie greoaie i zgomotoas (Fig. 3); edemul capului i gtului;
seroziti nazale i oculare; semne nervoase (torticolis, ataxie i lips de
coordonare a micrilor).
Au fost recoltate i expediate la Laboratorul Sanitar Veterinar Tulcea
probe de cadavre de gin, snge (Fig. 4) i tampoane cloacale (Fig 5), cu
suspiciunea de influen aviar.
n data de 14.10.2005, Institutul de Diagnostic i Sntate Animal
Bucureti a confirmat prezena virusului H5 n gospodria respectiv. Tot la
aceast dat a fost confirmat suspiciunea de grip aviar i la lebedele de var
din amenajarea piscicol Maliuc.

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Fig. 1. Cadavre ale ginilor gsite moarte n adpost

Fig. 2. Cianoza crestei i brbielor la o gain cu influen

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Fig. 3. Respiraie greoaie i cianoza crestei i brbielor

Fig. 4. Recoltarea probelor de snge

Fig. 5. Recoltarea tampoanelor cloacale

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CONCLUZII

Focarul de grip aviar declarat la gini n comuna Vulturu, judeul Tulcea


a aprut n urma contactelor psrilor domestice cu psrile migratoare. Acest
lucru s-a ntamplat n preajma luciului de ap limitrof, pe care au poposit lebede
de var la care s-a diagnosticat n acelai timp boala.
Au fost afectate ndeosebi psrile adulte, evoluia fiind de tip supraacut, cu
moarte subit n multe cazuri.
Principalele semne clinice ntlnite la psrile examinate au fost: edemul
capului i gatului , cianoza crestei i barbielor, seroziti nazale i oculare,
horiplumaie, inapeten, respiraie greoaie.
Diagnosticul s-a confirmat prin examen virusologic i RT-PCR de ctre
IDSA Bucureti, fiind identificat virusul influenei aviare, subtipul H5.

BIBLIOGRAFIE

1. Brougere-Picoux Jeanne, 2006 Le point sur linfluenza aviaire. Supplement


technique, nr. 97 a La Depeche Veterinaire.
2. Mnzat Moga R., 2001 - Boli infecioase ale animalelor. Ed. Brumar, Timioara.
3. Olaru E., 2005 - Instruciuni pentru recunoasterea Influenei aviare i diferenierea de
alte afeciuni ale psrilor. IDSA Bucureti.
4. Perianu T., 2005 - Boli infecioase ale animalelor. Viroze II. Editura Universitas XXI,
Iai.
5. Savua Gh., 2001 - Epidemiologie veterinar general. Ed. Ion Ionescu de la Brad
Iai.
6. *** Programul aciunilor de supraveghere, prevenire i control al bolilor la animale,
ANSVSA Bucureti, 2005.
7. *** O.I.E., Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, 5th
edition,2004, www.oie.int.

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ASPECTE EPIDEMIOLOGICE I CLINICE


N LISTERIOZA OVIN

C. CARP-CRARE, T. PERIANU, Elena VELESCU,


Irina-Oana TNASE, C. PAVLI, G. CIOBANU
U.S.A.M.V. Iai

In two ovine breeding units, between 2004 2005, epidemiological,


morphoclinical and therapeutical investigations, regarding ovine listeriosis
were made. From the total number of 1139 (692 from unit S and 447
from unit D) different breeds and ages ovine, 34 (2,98%) got ill, 8
(23,52%) of them died, 23 (67,66%) were sacrified by necessity and 3
(8,82%) have healed.
The ilnesses have occured in the cold season within live stock kept
in strict stabulation and fed preponderently with silo fodder.
The disease manifested clinically acute septicemicaly, in 7 (20,58%)
cases and with encephalomyelites manifestations in 27 (79,42%) cases.
The cultures made on usual cultivation mediums as well on blood
mediums from the different brain segments permetted the isolation of 10
Listeria monocytogenes strains. The strains were identified by their
morphological, cultural and biochemical characteristics and their
pathogenity was tested on mice and rabbits.
The treatment with ampyciline in dosis of 0,025mg/kg i.v. has led to
healing in 3 cases with septicemical acute evolutive forms.

Key words: Listeria monocytogenes, ovine, pathogenity, encephalomyelitis.

Incidena crescut a infeciilor cu listeria la cele mai variate specii de


animale, pierderile economice pe care le determin, polimorfismul manifestrilor
clinico-epidemiologice cu un procent ridicat de infecii inaparente sau atipice,
starea de purttor de lung durat, precum i pericolul pe care l prezint
animalele cu potenial infecios pentru om, au fcut ca listerioza s devin o
problem de mare actualitate att pentru patologia veterinar, ct i pentru
medicina uman; infecia fiind o zoonoz.
Incredinai de existena bolii i n aceast parte a rii, am socotit util s
investigm factorii epidemiologici, manifestrile clinice i posibilitile de
tratament, n condiii de teren, n dou focare de listerioz ovin.

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MATERIAL I METODE

Investigaiile s-au efectuat n perioada 2004-2005, n dou uniti (S i


D) de creterea ovinelor, cu un efectiv total de 1.139 (692 n unitatea S i
respectiv 447 n unitatea D) de rase i vrste diferite, unde s-au nregistrat
cazuri de mbolnvire. Pentru stabilirea diagnosticului s-au ntreprins investigaii
epidemiologice, morfoclinice i de laborator.
Confirmarea diagnosticului s-a fcut pe baza examenului bacteriologic. n
vederea izolrii germenului s-au fcut nsmnri pe medii de cultur din
organele parenchimatoase i din mai multe zone ale encefalului, de la 10 ovine
care au murit i care n via prezentau manifestri ce suspiciona listerioza.
Au fost examinate n aceast direcie 16 probe de la 10 ovine, provenind
din cele dou uniti, izolndu-se un numr de 10 tulpini de L. monocytogenes.
Controlul patogenitii tulpinilor a fost fcut prin inoculri la oareci a
culturilor de 24 ore din bulion. oarecii inoculai au murit dup 4-7 zile,
prezentnd n mod constant leziuni de hepatit necrotic din care au fost izolate
de fiecare dat listerii.
Instilarea conjunctival a culturii de 24 ore la iepure a produs o
conjunctivit acut seroas, transformat apoi n purulent.

REZULTATE I DISCUII

Primul focar de listerioz din unitatea S a fost identificat la sfritul lunii


ianuarie 2004, ntr-o turm de 692 ovine. n decurs de 15 zile, n perioada 29
ianuarie 13 februarie 2004, s-au mbolnvit 18 oi, ceea ce reprezint 2,60%.
Din cele 18 ovine mbolnvite 6 (33,33%) au murit , 9 (50,00%) au fost
sacrificate de necesitate, iar 3 (16,67%) s-au vindecat 3 (16,67%).
mbolnvirile au aprut n urma introducerii n alimentaia oilor, a
porumbului nsilozat provenit dintr-un siloz proaspt nceput. Aceasta a dus la
bnuiala c furajul nsilozat a constituit factorul favorizant n apariia
manifestrilor de boal. Mai mult, afirmaia se sprijin i pe faptul c n urma
schimbrii alimentaiei prin eliminarea furajului murat i a introducerii
concentratelor n raie, nu au mai fost nregistrate cazuri de mbolnvire.
Al doilea focar de listerioz a fost confirmat bacteriologic n perioada
februarie-martie 2005, ntr-o alt turm de ovine, cu un efectiv de 447. De la 21
februarie i pn la 26 martie 2005, ntr-o perioad de 34 de zile, s-au mbolnvit
16 ovine, reprezentnd 3,57%. Din cele 16 ovine cu simptome de boal 2
(12,50%) au murit iar 14 (87,50%) au fost sacrificate de necesitate. Vindecri nu
s-au nregistrat, cu toate c oile bolnave au fost tratate cu antibiotice (ampicilin).
Turma de ovine a primit n raie, pe lng alte sortimente i furaj nsilozat.
Apariia primelor cazuri de mbolnvire i n aceast unitate, a coincis cu
administrarea n hran a silozului de lucern, nou nceput. mbolnvirile au ncetat
brusc, fr a se lua msuri privind mbuntirea condiiilor de igien, alimentaie
i fr aplicarea unor msuri speciale de profilaxie i combatere.

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Din investigaiile epidemiologice ntreprinse n cele dou uniti rezult c,


din totalul de 1139 ovine s-au mbolnvit 34 (fig. 1), ceea ce reprezint 2,99%, au
murit 8 (23,52%), au fost sacrificate de necesitate 23 (67,66% ) i s-au vindecat 3
(8,82 %).

Fig. 1 Rezultatele investigaiilor epidemiologice n cele dou uniti cu listerioz la ovine

mbolnvirile au aprut numai n sezonul rece, la efective inute n


stabulaie strict i numai pn la nceperea punatului. De menionat, c n cele
dou focare examinate nu s-au nregistrat avorturi, dei cazurile de boal au
aprut n a doua jumatate a perioadei de gestaie a oilor.
De asemenea, din aceleai investigaii rezult c, boala a aprut sub form
enzootic sezonier, n lunile ianuarie aprilie, cu o morbiditate cuprins ntre 2,6
i 3,57%, foarte apropiat de cea descris i n literatura de specialitate. ntruct
odat cu mbuntirea factorilor de microclimat (creterea temperaturii mediului
ambiant), ntroducerea n alimentaie a furajului concentrat i scoaterea animalelor
la pune, nu s-au mai semnalat alte cazuri de mbolnvire ne face s considerm
c, la apariia bolii au contribuit unii factori climatici (umezeala, frigul),
deficienele de alimentaie i ali factori de ordin igienic.
La apariia bolii, n mare msur a contribuit i alimentaia ovinelor, care,
n special n a doua perioad a iernii, a fost deficitar att cantitativ, ct i
calitativ, folosindu-se excesiv furaje nsilozate, unele mucegite, n majoritate
fibroase i n cantiti insuficiente.
Aspectul clinic al bolii n focarele descrise a fost aproximativ acelai,
corespunznd datelor din literatura de specialitate (fig. 2). Astfel din cele 34 ovine
cu manifestri de listerioz la 7 (20,58%) evoluia a fost acut septicemic, iar 27
(79,42%) ovine au prezentat simptome de encefalomielit, evoluie subacut. De

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menionat c nu s-au nregistrat nici un caz de avort listeric. De fapt n literatura


de specialitate se afirm c, localizarea genital exprimat prin avort se ntlnete
rar, de obicei la oile primipare i niciodat nu evolueaz concomitent cu
manifestrile de meningoencefalit (forma nervoas). De asemenea, menionm
c formele evolutive acute, septicemice, au fost diagnosticate numai la mieii
sugari i la tineretul ovin.

Fig. 2. Forme evolutive n listerioza ovin

La ovinele mbolnvite s-a observat de la nceput o stare de abatere,


reducerea total a poftei de mncare, sau chiar dispariia apetitului. Respiraia era
accelerat, iar temperatura corporal varia n limite mari, nregistrndu-se o
hipotermie, n faza iniial, revenind aproape n limitele normale n ultima
perioad a evoluiei bolii. n unele cazuri s-au semnalat conjunctivite, iar unele
exemplare prezentau scurgeri seroase din cavitile nasale, altele salivaie
abundent (fig. 3).

Fig. 3. Aspect clinic salivaie


abundent; pedalri.

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Majoritatea oilor mbolnvite au prezentat manifestri clinice din partea


sistemului nervos central, reprezentate prin dromomanie, vertije, opistotonus
(fig. 4).

Fig. 4. Aspecte clinice (opistotonus, devierea lateral a capului)

Tulburrile nervoase au progresat concomitent cu evoluia n timp a bolii.


Ulterior au aprut paralizii ale membrelor, oile bolnave nu se mai puteau scula,
iar ridicate de jos cadeau. n majoritatea cazurilor oile stteau n decubit lateral i
executau cu picioarele micri de pedalare. Deseori s-au observat tulburri
oculare manifestate prin amauroz.
nsmnrile efectuate pe medii obinuite de cultur, ca i pe medii cu
snge (agar cu snge), din organele parenchimatoase (ficat, splin ) ca i din
diverse segmente ale encefalului de la 10 cadavre au dus la izolarea i
identificarea a 10 tulpini de Listeria monocytogenes. Germenii s-au izolat mai
frecvent de la nivelul bulbului n toate cazurile (100%), apoi din tuberculii
cuadrigemeni (80%) i pedunculii cerebrali i cerebeloi (70%). ncercrile de
izolare din organele parenchimatoase (ficat i splin) i din snge, chiar la
animalele de la care Listeria monocytogenes s-a izolat din bulb, au fost negative.
Biochimic, tulpinile de Listeria monocytogenes izolate au produs indol i
hidrogen sulfurat; au fermentat glucoza, maltoza i dextrina; erau mobile i au
hemolizat globulele roii de iepure i de oaie.
Inoculrile celor 10 tulpini, culturi n bulion glucozat, de 24 de ore n doz
de 0,2 ml pe cale intraperitoneal au determinat moartea oarecilor albi n decurs
de 4-7 zile. Din cele 10 tulpini, una izolat de la un miel s-a dovedit slab
patogen.
Instilat n sacul conjunctival la iepure, listeria a determinat n decurs de 2-
3 zile o conjunctivit de tip granular i o cheratit difuz. Procesul inflamator
ocular s-a resorbit n cteva zile, dar animalul a rmas cu opacitatea permanent a
corneii.

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Toate cele 34 de ovine cu simptome de listerioz au fost supuse


tratamentului cu ampicilin n doz de 0,025 mg/kg, pentru 24 de ore. Inocularea
s-a fcu pe cale intravenoas. Din cele 34 de animale tratate, numai n 3 cazuri s-
au obinut vindecri. De menionat, c cele trei vindecri s-au obinut la animalele
(miei i tineret) cu forme evolutive acute, septicemice. Nici un animal cu
manifestri nervoase, nu s-a vindecat ca urmare a tratamentului aplicat.

CONCLUZII

Investigaiile epidemiologice, morfoclinice i de laborator n dou episoade


de listerioz ovin au condus la desprinderea urmtoarelor concluzii:
1. mbolnvirile s-au nregistrat n perioada ianuarie-aprilie, n condiii de
stabulaie strict i prelungit, cu o alimentaie n care predomina furajul nsilozat.
2. Din totalul de 1.139 ovine, s-au mbolnvit 34, ceea ce reprezint 2,99%,
iar din acestea 8 (23,52%) au murit, 23 (67,66%) au fost sacrificate de necesitate,
iar 3(8,82%) animale s-au vindecat.
3. Clinic boala a evoluat acut septicemic n 7(20,58%) cazuri, prezente
numai la tineretul ovin(miei i tineret) i subacut cu manifestri nervoase la 27
(79,42%) de ovine.
4. n evoluia subacut cu manifestri din partea sistemului nervos s-a
nregistrat dromomanie, vertije, opistotonus, pareze i paralizii ale buzelor,
membrelor, decubit i moarte.
7. De menionat c nu s-au nregistrat avorturi la oile gestante.
8. nsmnrile pe medii de cultur cu snge i pe medii glucozate, din
diverse zone ale sistemului nervos central, din organele parenchimatoase (ficat i
splin) i din snge au dus la izolarea a 10 tulpini de Listeria monocytogenes,
identificate pe baza caracterelor morfologice, culturale, biochimice i de
patogenitate.
9. Determinarea patogenitii tulpinilor izolate s-a fcut prin inocularea
culturilor de 24 de ore n bulion, att la oricei albi ct i prin instilare n sacul
conjunctival la iepure.
10. Tratamentul cu ampicilin n doz de 200 mg/kg i zi administrat pe
cale intravenoas a dus la obinerea videcrii la 3 animale cu manifestri acute
septicemice de boal. Nu s-a nregistrat nici o vindecare la animalele cu
manifestri din partea sistemului nervos.

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BIBLIOGRAFIE

1. Larpent J.P., 2000 - Listeria, Ed. Tec.DOC, 2e Edition, Paris.


2. Marly J., Heuchel V., Ribaud D., 2004 - Evaluation de l'efficacit des conservateurs
sur la prvention du dveloppement de Listeria monocytogenes dans les
ensilages. 11me Rencontres autour des Recherches sur les Ruminants, Paris
(FRA).
3. Moga-Mnzat R., (colaborator), 2001 Boli infecioase ale animalelor. Bacterioze.
Editura Brumar, Timioara.
4. Perianu T., tefan N., CarpCrare M., Velescu Elena, Alexandru N., uc C.I.,
2003 Boli infecioase ale animalelor. Bacterioze. Vol. I. Editura Venus, Iai.
5. Rpuntean Gh., Rpuntean S., 1999 - Bacteriologie special veterinar. Ed. Tipo-
Agronomia, Cluj-Napoca.
6. Zundel E, Bernard S., 2004 - In vivo imaging of the spread of Listeria
monocytogenes in sheep. 15th International Symposium on Problems of
Listeriosis (Isopol), Uppsala (SWE).

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OBSERVATIONS REGARDING THE ISOLATION OF


LISTERIA MONOCYTOGENES IN SILO FODDER
C. CARP-CRARE
U.S.A.M.V. Iai

The researches have followed the isolation of Listeria


monocytogenes from the silage. The silage samples were taken from a ovine
breeding unit, where listeriosis cases have been diagnosticated. In the
examined unit the main fodder was the silage, because of the indoor
breeding periode during the cold season (February -March).
From the 42 analysed silage samples there were isolated 3 (7,14%)
Listeria monocytogenes stems. Two (4,76%) from the three identified stems
have been isolated from the silage surface and one of them (2,38%), from
the edges of the silage platform. It is important to mention that from the
silage samples with the pH value below 5, there hasn't been isolated any
Listeria monocytogenes stem.

Key words: Listeria monocytogenes, silage, ovine.

Irrational alimentation, specially with silage excess, of inadequate quality,


incomplete fermented, with a pH over 5,5, is the main factor in disease starting
and the rason why listeriosis was called "silo disease" as well. Also, often, in
listeriosis focuses, after the improvement of some alimentary deficiencies, the
diseases reduction or cessation is observed.
In ovine, the using of inadequate quality silage and in excesive quantities
represents one of the main factors of infection susceptibility raising.
Its may be said that alimentation has a favourable role, sometimes even
decisive in disease staring and that the infections with Listeria monocytogenes
have a very powerfull conditioned character.

MATERIAL AND METHOD

The researches were executed on a number of 42 samples obtained from a


unit with listeriosis cases in sheeps which had silage as a basic alimentation,
because of the indoor breeding periode.
The samples were obtained from different zones of the silage, the number
of prelevated samples being in function of the pH. Because the pH in silo depth
was under 5, pH which not allows the listeria multiplication, was choosed a small
samples number, respectively 6, while the surface and the edges of the silage
platform ph varied between 5,5 and 6, being prelevated a number of 18 samples
(table 1).

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Table 1
Silage samples sourse

Total sample Surface samples Edges samples Depth samples


number (pH-6) (ph 5,5-6) (ph<5)
42 18 18 6

50
42
40

30

20 18 18

10 6

0
Total sample Surface Edges samples Depth samples
number samples (pH-6) (ph 5,5-6) (ph<5)

Fig. 1- Silage samples sourse

Thesamples were prelevated as steril as possible and were complied to


microbiological control for listeria isolation.
In keeping with to some score system for silage quality, in function of pH it
may be observed that the analysed samples didnt fulfilled the characteristics of
some good quality silage, having 2 from a maximum of 5 points (table 2).
Table 2
The score system of silage quality on a base of pH value

Ph value Ph indicating colour Score


4,2 Red 5
4,2-4,6 Red - orange 4
4,6-5,1 Orange 3
5,1-6,1 Yellow 2
6,1-6,4 Yellow greenish 1
6,4-7,2 Green 0

As a work technics was used a standard isolation technics (Centre National


d'Etudes Vtrinaires et Alimentaires-CNEVA Pr 116/2000) specially recomanded
for silage, very similar with the one applied in our country.
In keeping with this method, from the samples brought in the laboratory
were weight 25 gramme of silage, which were introduced in pre-enrichment demi
Fraser broth. The samples were left for incubation 24 hours at 300C.
After incubation periode a sowing on selective Palcam and Oxford culture
mediums was made and 0,1 ml from pre-enrichment broth was transferred in

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enrichment Fraser broth, which was incubated 36-48 hours at 370C. From those
ones sowing in 24 as well as in 48 hours on Palcam and Oxford selective culture
mediums were realized, the plates being incubated and observed for two days
long.
The suspect colonies (Palcam gray greenish center surrounded of a
black halo and on Oxford-black colonies surrounded of a black halo) were
transplanted on selective culture mediums such as TSYA agar (triptonis, soya)
with ram blood, in order to observe haemolysis. The plates were incubated at
370C for 24 hours and then, the specific colonies were submitted to minimum
identification tests (Api-Listeria, CAMP).
The stems pathogenity was observed in vitro using strains cropping on ram
blood agar, as well as in vivo, inoculating little laboratory mice.

RESULTS AND DISCUTIONS

After the researches for Listeria monocytogenes isolation in fodders, the


fact that from a total number of 42 analysed silage samples, were isolated 3
(7,14%) Listeria monocytogenes strains was observed. From the 3 identified
strains, 2 (4,76%) were isolated from the silage surface, 1 strain (2,38%) from the
edges of the silage platform, while in depth samples there hasn't been isolated any
Listeria monocytogenes stem (table 3).
For the bacteria isolation from the silage samples, Palcam and Oxford
mediums have been proved eficient and intensely selective. The identification was
realized using 2 tests (CAMP and Api-Listeria) which allow Listeria
monocytogenes difference from other genus species, considered unpathogenes.
The pathogenity of isolated stems was proved inoculating little mice.. After
some fresh culture centrifugation, from the obtained sediment the culture was
reconstituted in 1 ml physiological serum. After this the mice were intraperitoneal
inoculated with 0,1 ml reconstituded suspension, their death happening between 7
-10 days, all 3 isolated strains being proved to be pathogenes.
It is important to mention that Listeria monocytogenes stems isolation from
the silage was realized during its administration as a basic alimentation and in the
same time with some listeriozis cases appearance in some adult sheeps, with
nervous signs.
An important aspect is represented by the fact that once the silage changing
is made, the isolated listeriozis case disapeared.
The pH over 5 present in some silage areas had a decisive role in bacteria
excessive developement. This aspect, which follows from our reseach too, by the
fact that Listeria couldnt be isolated from the silage depth where the pH
was under 5, but only from the samples where pH were over 5, argues the
statement that listeriosis in sheeps is a disease owed to alimentation deficiencies,
specially bad quality silage ("silo disease").

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Table 3
Listeria monocytogenes frequency in silage

Positive Negative
Samples Samples number
pH samples samples
sourse
No. % No. % No. %
Surface
1. 6 18 42,86 2 4,76 16 38,1
samples
Edges
2. 5,5-6 18 42,86 1 2,38 17 40,48
samples
Depth
3. <5 6 14,28 - - 6 14,28
samples
4. Total - 42 100 3 7,14 39 92,86

50
42,86 42,86
40,48
40 38,1

30

20 14,28 14,28

10 4,76
2,38
0
0
Samples number Positive samples Negative samples

Surface sam ples Edges sam ples Depth sam ples

Fig. 3 - Listeria monocytogenes frequency in silage

CONCLUSIONS

The analyses regarding Listeria monocytogenes presence in silage, brought


the following conclusions:
1. In the silage Listeria monocytogenes was isolated in 7,14% from the total
of the 42 analysed samples.
2. From the 3 isolated strains, 2 (4,76%) were taken from the surface and
one (2,38%) of the platform edges.
3. In depth samples, where the pH was under 5 it has been izolated no
Listeria monocytogenes strain.

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4. Listeria monocytogenes isolation from the silage a coincided with some


isolated listeriosis cases appearence.
5. All the isolated stems were proved to be pathogenes by intraperitoneal
inoculation of little mice, using fresh culture, the inoculated quantity being of 0,1
ml.

BIBLIOGRAPHY

1. Gitter M., Stebbings R.S., Morris J.A., Hannam D., Harris C., 1986 - Relationship
between ovine listeriozis and silage feeding. Vet. Rec., 118 (8), 207-208
2. Larpent J.P., 2000 - Listeria, Ed. Tec.DOC, 2e Edition, Paris
3. Perianu T., 2003 - Boli infecioase ale animalelor domestice, Bacterioze, Vol.I, Ed.
Venus-Iai, p. 429-436
4. Rpuntean Gh., Rpuntean S., 2005 - Bacteriologie veterinar special, Ed.
Academic Pres, Cluj-Napoca, p.38-46
5. Y. Milleman, D. Remy, J. Brugere-Picoux., 2000 - La listeriose des ruminants-
etiologie, pathogenie et epidemiologie. Point Vet., vol. 31, nr. 208, 313-316

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DIAGNOSTIC DE LABORATOR COMPLEX AL


TUBERCULOZEI LA BOVINELE SACRIFICATE
THE COMPLEX DIAGNOSIS OF TUBERCULOSIS INFECTION
AT THE SACRIFICED CATTLE

Adriana CHIRIAC1, M. CARP- CRARE2,


C. NECULI
1
D.S.V.S.A. Vaslui
2
U.S.A.M.V. Iai

Bovine tuberculosis infection is usually diagnosed by post-mortem


examination and histopathological and bacteriological techiques.
Traditional bacterial culture remains the routine method for confirmation
of infection.
Bacteriological examination may comprise the demonstration of
acid-fast bacilli by microscopic examination, and the isolation of
mycobacteria on selective culture media and their subsequent identification
by cultural and biochemical tests.
Animal inoculation is slightly more sensitive than culture, but should
be used when histopathology lesions are compatible with mycobacteriosis
infection and when isolation in culture is negative.
During 2000-2005, 69 samples were processed within Sanitary
Veterinary Laboratory Vaslui for necropsy, bacteriological examination,
histhopatological examination and animal inoculation.
We have examinated 21 samples through microscopic examination
and the isolation of mycobacteria on selective culture (Lweinstein -
Jensen).
The necropsy and the bacteriological examination were positive at
11 samples (13,34%), all the samples beeing confirmed by
histhopatological examination.

Key words: diagnosis, mycobacteria, bacteriological and histopathological


examination.

MATERIALE I METODE

n perioada 2000-2005, n Laboratorul Sanitar Veterinar Vaslui s-au


examinat 69 de probe n vederea examenelor anatomopatologice, bacteriologice,
histopatologice i a bioprobei pe cobai n direcia tuberculozei bovine. Dintre
acestea, 50 de bovine au fost tuberculino- negative.
De asemenea, 15 probe limfonoduri i organe au provenit de la bovine
reacionate pozitiv la testul EIAs- IFN.

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Din totalul animalelor sacrificate pentru stabilirea diagnosticului de


tuberculoz, s-au primit la laborator 29 probe poriuni de organe (ficat, splin,
rinichi, pulmon, aparat genital).
Examenul bacteriologic a fost efectuat prin 3 metode astfel:
- examen bacterioscopic pe un numr de 735 frotiuri colorate prin
metoda Ziehl- Neelsen att din cazeum, ct i din zona periferic a
nodulului tuberculos;
- examen bacterioscopic n lumin fluorescent pe un numr de 27
frotiuri colorate cu substane fluorocrome (auramina-rodamina);
- examen cultural s-a efectuat la un numr de 21 probe: 4 probe
provenite din pulmon i limfonodurile aferente; 15 probe provenite din
limfonodurile capului, toracelui, mezenterici, hepatici, iliaci i din ficat,
splin, rinichi, pulmon, aparatul genital; 2 probe din leziuni de la cobaii
cu bioproba pozitiv.
nsmnrile s-au fcut pe mediul Loweinstein Jensen cu i fr
glicerin.S-au nsmnat pentru fiecare prob cercetat cte 4 tuburi cu mediu
Lowenstein - Jensen (2 cu glicerin i 2 fr glicerin). Culturile au devenit vizibile
dup a 50-a zi de la nsmnare. Coloniile au prezentat o cretere disgonic, de
tip S, avnd suprafaa i marginile netede i lucioase, nepigmentate, cu aspect
uor bombat, de obicei mici i izolate (tabelul 2). S-au dezvoltat mai bine pe
mediile fr glicerin i n cele neaerate.
Testele biochimice folosite pentru identificarea micobacteriilor au fost:
testul catalazei, testul nitratreductazei i testul niacinei.
Metodele de colorare a seciunilor de esuturi etalate pe lam au fost:
hematoxilina eozina - albastru de metil (HEA) i Ziehl- Neelsen modificat.

REZULTATE I DISCUII

Examen anatomopatologic
Rezultatele examenului anatomopatologic sunt sintetizate n tabelul 1.
Tabelul 1
Leziunile limfonodulare de la bovinele examinate n laborator n perioada 2000-2005

Specificare 2000 2001 2002 2003 2004 2005 Total Procent


Probe analizate pentru
diagnosticul de laborator al 19 18 14 5 9 4 69 100
tuberculozei bovine
Probe negative pentru
19 16 9 3 7 4 58 84,06
tuberculoz bovin
Probe pozitive
0 2 5 2 2 0 11 13,34
pentru tuberculoz bovin
Alte leziuni Hiperplazii
0 1 1 2 1 0 5 6,84
anatomo- limfoide
patologice Focare de
limfonodulare antracoz 0 5 1 1 0 4 11 15,06

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Leziuni suspecte de tuberculoz s-au constatat doar la 11 animale, care n


final au fost confirmate prin alte examene de laborator. Leziunile au fost sub
form de focare pulmonare acino-nodoase, cenuii-glbui, nconjurate de esut
pulmonar sclerozat (fig. 1).

Fig. 1 Focare cazeo-calcaroase


tuberculoase pe seciune n
limfonodurile mediastinale

Examenul bacteriologic
Examenul bacterioscopic a fost pozitiv la 11 din cele 69 de probe analizate
(13,34 %), toate probele fiind confirmate prin examenul histopatologic. Are o
valoare de diagnostic redus n tuberculoza bovin, datorit faptului c leziunile
sunt de obicei paubacilare .
n urma examenului bacteriologic s-au izolat 11 tulpini bacteriene cu
urmtoarele caractere culturale i morfologice (tabelul 2):

Tabelul 2
Identificarea tulpinilor micobacteriene

Timp de
Nr. Tip Examen macroscopic Examen microscopic
incubaie
crt. cultural al culturii al culturii
(zile)
1. 55 DISGONIC S Colonii cu margini neregulate, Bacili acidorezisteni
opace, alb-glbui,2-4 mm, cu aezai n grmezi
rofizare ulterioar neregulate.
2. 59 DISGONIC S Colonii cu margini ondulate, Bacili acidorezisteni
opace, conopidiforme,glbui, aezai n grmezi
2-4 mm, cu rofizare neregulate.
ulterioar
3. 60 DISGONIC S Colonii rotunde, opace, alb- Bacili acidorezisteni
glbui, 3-4 mm, cu rofizare aezai n grmezi
ulterioar neregulate.

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Timp de
Nr. Tip Examen macroscopic Examen microscopic
incubaie
crt. cultural al culturii al culturii
(zile)
4. 60 DISGONIC S Colonii cu margini neregulate, Bacili acidorezisteni
opace, glbui, bombate, 2-3 aezai n grmezi
mm, cu rofizare ulterioar neregulate.
5. 65 DISGONIC S Colonii rotunde opace, alb- Bacili acidorezisteni
glbui,3-4 mm, cu rofizare aezai n grmezi
ulterioar neregulate.
6. 55 DISGONIC S Colonii crateriforme, opace, Bacili acidorezisteni
glbui, 3-5 mm, cu rofizare aezai n grmezi
ulterioar neregulate.
7. 60 DISGONIC S Colonii rotunde, opace, alb- Bacili acidorezisteni
glbui, 3-4 mm, cu rofizare aezai n grmezi
ulterioar neregulate.
8. 60 DISGONIC S Colonii cu margini neregulate, Bacili acidorezisteni
opace, glbui, bombate, 2-3 aezai n grmezi
mm, cu rofizare ulterioar neregulate.
9. 65 DISGONIC S Colonii rotunde opace, alb- Bacili acidorezisteni
glbui,3-4 mm, cu rofizare aezai n grmezi
ulterioar neregulate.
10. 55 DISGONIC S Colonii crateriforme, opace, Bacili acidorezisteni
glbui, 3-5 mm, cu rofizare aezai n grmezi
ulterioar neregulate.
11. 65 DISGONIC S Colonii rotunde opace, alb- Bacili acidorezisteni
glbui,3-4 mm, cu rofizare aezai n grmezi
ulterioar neregulate.

Teste biochimice folosite pentru identificarea micobacteriilor


S-a observat la examinarea caracterelor biochimice a unui numr de 11
tulpini izolate la examenul cultural c 6 tulpini (54,55%) au fost catalazo-pozitive,
5 (49,45%) tulpini au fost catalazo-negative, toate cele 11 tulpini nu au redus
nitriii i au dat un rezultat negativ la testul niacinei (tabelul 3).
Tabelul 3
Rezultatele examenelor biochimice
(testul catalazei, nitratreductazei i a niacinei)

Nr.crt. Catalaza Nitratreductaza Testul niacinei


1. + - -
2. - - -
3. + - -
4. + - -
5. - - -
6. - - -
7. + - -
8. - - -
9. + - -
10. + - -
11. - - -

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Examenul histopatologic
Prin coloraia HEA s-a efectuat un examen de ansamblu al modificrilor
citologice, cu evidenierea structurii histopatologice a granulomului tuberculos,
fcndu-se n acelai timp i examen diferenial fa de alte granuloame
nespecifice (actinobacilar, micotic, de corpi strini etc.).
Metoda de colorare Ziehl-Neelsen modificat a permis evidenierea
structurii histopatologice a esutului i a bacililor acido- alcoolorezisteni.Prin
aceast metod de colorare s-au putut evidenia i bacilii acido-alcoolorezisteni
prezeni n citoplasma celulelor epitelioide, gigante i n zona de necroza a
granulomului (fig. 2).

Fig. 2 Limfonod bovine:


proliferare epitelioid. Coloraia
Ziehl-Neelsen modificat.

Bioproba
Bioproba s-a efectuat pentru cele 69 probe primite n laborator n perioada
analizat.
La 7 probe recoltate de la animale tuberculino-negative, nu s-a efectuat
bioproba pe cobai, ntruct rezultatele examenelor: anatomopatologic,
bacterioscopic i histopatologic au fost negative pentru tuberculoza bovin.

Studiu statistic al activitii de supraveghere bovin din judeul Vaslui


prin examene de laborator
n perioada 2000-2005, s-au sacrificat pentru precizarea diagnosticului un
numr de 57 de bovine tuberculino-reagente (3,03%). Din cele 69 de probe
analizate, boala s-a confirmat la 11 capete, reprezentnd 13, 34% procent mediu
de contaminare pe ntreaga perioad de studiu.
S-a constatat o rat de confirmare de 0,11% ( 2/18) n 2001, de 0,35%
(5/14) n 2002, de 0,50% (2/5) n 2003 i de 0,22% (2/9) n 2004.
Din analiza rezultatelor acestor examene, se poate aprecia sensibilitatea i
specificitatea testelor de diagnostic pe animale sacrificate.

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Ss = a/ a+c% = 11/ 11 + 3 % = 78,57%


Sp= d/ d+b% = 59/59 + 10 % = 85,51%
a = real pozitive;
b = fals pozitive;
c = fals negative;
d = real negative;
6

5 5,0

4
Numr probe pozitive

2 2,0 2 2

0 0,0
0 0 0 0,0 0,0 0,0
0
2000 2001 2002 2003 2004 2005
Perioada
Numrul probelor pozitive din colectiviti
Numrul probelor pozitive din gospodriile populaiei

Graficul 1- Situaia probelor pozitive prin examen de laborator n perioada 2000-2005

CONCLUZII

ntre anii 2000-2005, s-au primit n laborator probe de la un numr de 69


bovine, care au fost analizate pentru stabilirea diagnosticului de tuberculoz
bovin. Din acestea, 19 probe proveneau de la animalele tuberculino-reagente i
50 probe de la animale tuberculino-negative.
La examenul anatomo-patologic efectuat n laborator la marea majoritate a
limfonodurilor provenite de la animalele tuberculino-reagente, care au fost n final
confirmate, s-au decelat leziuni caracteristice de dimensiuni i aspecte diferite.
Din totalul animalelor sacrificate pentru stabilirea diagnosticului de
tuberculoz bovin, s-au primit n laborator 29 probe de organe, la care s-au
constatat leziuni susupecte de tuberculoz la 11 probe, la care n final boala a fost
confirmat.
Examenul bacterioscopic a fost pozitiv la cele 11 probe confirmate. Dei
acest examen presupune examinarea unui numr mare de cmpuri microscopice,
valoarea sa de diagnostic asigur susinerea acestuia.

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Au fost analizate prin examen cultural 21 de probe, din care 11 au fost


pozitive, fiind confirmate n final i prin bioprob.
S-au examinat histopatologic toate cele 69 de probe primite n laborator.
Dintre acestea, la 11 probe cu diagnostic de tuberculoz bovin confirmat s-au
decelat constant detaliile morfopatologice patognomonice bolii.
Avnd n vedere valoarea de diagnostic a examenului histopatologic prin
utilizarea metodei de colorare Ziehl-Neelsen modificat, se sugereaz propunerea
ca probele cu leziuni patognomonice n care se deceleaz bacilii acido-
alcoolorezisteni, s aib valoare decizional n stabilirea diagnosticului, fr a
mai fi necesar bioproba, reducnd astfel perioada de emitere a buletinului de
analiz.
Bioproba s-a efectuat pentru toate probele primite n laborator.
Dintr-un numr de 29 probe organe, care au nsoit probele de limfonoduri,
20 probe (68,57%) prezentau leziuni parazitare, n special de echinococoz, fapt
care ar putea explica o parte din reaciile nespecifice ntlnite la examenul alergic.

BIBLIOGRAFIE SELECTIV

1. Alexandru N., 1994 - Tehnic simpl, rapid de prelevare i colorare a elementelor


citopatologice n tuberculoza limfonodal, Al VI-lea Congres de Medicina
Veterinara, 63
2. Bercea I., Popescu t., Sandu I., 1999 - Diagnostic alternative proceedings of cattle
tuberculosis in Romania, Proceedings 26 th World Veterinary Congress WSA,
Lyon, France, CD
3. Carp - Crare M., Eleonora Guguianu, Dorina Timofte, 1997 - Lucrri practice de
microbiologie veterinar, Uz intern, U.A.M.V. Iai
4. Corner L.A., 1994 - Past mortem diagnosis of Mycobacterium bovis infection in
cattle, Veterinary Microbiology , 40, 53-63
5. Dumitrache Mihaela, 2005 - Cercetri asupra diagnosticului de supraveghere i de
confirmare - infirmare a tuberculozei bovine n municipiul Bucureti i n judeul
Ilfov, Tez de doctorat, Universitatea de tiine Agronomice i Medicin
Veterinar Bucureti, Bucureti, pag. 175- 186
6. Ionic C., Grigore C., Leonte I., eveloiu I., 1988 - Localizarea leziunilor
macroscopice de tuberculoz la taurine infectate, Simp.,, Tuberculoza la om i
animale, Slobozia, 14
7. Ni C., 2006 - Cercetri privind epidemiologia, diagnosticul i profilaxia tuberculozei
bovine n judeul Vaslui, tez de doctorat, Universitatea de tiine Agricole i
Medicin Veterinar Bucureti
8. One E., Virginia Constantinescu, 1978 - Diagnosticul de laborator n medicina
veterinar, Ed. Ceres, Bucureti, 74-78
9. Popescu t., Sandu I., 1997 - Preocupri i perspective n domeniul diagnosticului,
prevenirii i combaterii tuberculozei bovine, Ed. L.C.S.V.D. Bucureti, pag. 5-33
10. Rducnescu H., Bica-Popii Valeria, 1986 Bacteriologie veterinar, Editura
Ceres, Bucureti
11. *** Diagnosticul de laborator al tuberculozei la animale - Instruciuni MAIA nr.80296/
1984
12. *** Manual O.I.E.(2004) - Manual of Diagnosis Tests and Vaccines for terrestrial
Animals, part 2, section 2.3., chapter 2.3.3.

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SINDROMUL DERMATITEI SI NEFRITEI PORCINE


(PDNS) O NOU ENTITATE MORBID N PATOLOGIA
SUIN DIN ROMNIA
PORCINE DERMATITIS AND NEPHROPATHY SYNDROME (PDNS) A NEW
PATHOLOGICAL ENTITY IN ROMANIAN SWINE HERDS

D. CADAR1, Marina SPNU1, V. MICLU1,


L. KBLKUTI1, T. TUBOLY2, G. CZIRJK1,
A. DN3, A. CSGOLA2
1
U.S.A.M.V. Cluj Napoca
2
Faculty of Veterinary Sciences, Budapest, Hungary
3
Central Veterinary Institute, Budapest, Hungary

PDNS is a relatively new and often fatal disease that primarily


affects recently weaned and feeder pigs from 1.5 to 4 months of age (Smith
et al., 1993; Thibault et al., 1998). The syndrome was first recognized in the
UK in 1993 (Smith et al., 1993). PDNS is generally sporadic. Pigs affected
with PDNS are older pigs, particularly animals ranging from 12 to 14
weeks of age, but PDNS has also been described in finishing pigs and
replacement gilts. The first signs were skin lesions that were multifocal,
well circumscribed, slight raised, dark red, circular to irregular and 120
mm in diameter. The pathological changes are consistent with an immune-
complex disorder,with severe renal lesions usually causing fatal azotaemia.
Porcine circovirus type 2 (PCV2), believed to be an important cause of
PDNS. Significant gross lesions are present consistently in the skin and
kidneys of swine with PDNS, although other organs may also be affected
(Choi and Chae, 2001; Duran et al., 1997; Thibault et al., 1998). Gross
skin lesions consist of several round to irregularly shaped red to purple
macules and papules that coalesce over the perineum and distal limbs to
form large irregular patches. The kidneys are enlarged and have pale
cortices with multiple red circular haemorrhagic cortical foci measuring 2
4 mm in diameter. Renal and inguinal lymph nodes are usually enlarged
and red (Ramos-Vara et al., 1997). The characteristic microscopic lesions
of PDNS are generalized severe necrotizing vasculitis and fibrinonecrotic
glomerulonephritis (Choi and Chae, 2001; Choi et al., 2002; Duran et al.,
1997; Ramos-Vara et al., 1997). This paper described the first PDNS cases
from romanian swine herds. Porcine circovirus type 2 was detected in
kidneys and other tissues by in situ hybridization and polymerase chain
reaction (PCR).

Cunoaterea principalelor semne clinice ct i a celor anatomopatologice


n sindromul dermatitei i nefritei porcine (PDNS) este foarte important att n
direcia diagnosticului pozitiv, ct i n diagnosticul diferenial, existnd

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numeroase semne clinice i anatomopatologice care se pot confunda cu cele din


alte boli, cum ar fi cele din pesta porcina clasic, rujet.
Scopul acestui studiu a fost, evidenierea, respectiv prezentarea
principalelor manifestri clinice si anatomopatologice ale sindromului dermatitei
i nefritei porcine (PDNS), precum i diagnosticul acestui sindrom i prin acesta
diagnosticul circovirusului porcint tip 2 (PCV2).

MATERIAL I METOD

Acest studiu cuprinde observaiile nregistrate din perioada martie 2003


ianuarie 2006 n cteva ferme din centrul i nord-vestul Transilvaniei. Avnd n
vedere faptul c prevalena acestui sindrom n fermele de suine este cuprins
ntre 0,001- 0,03%, pe perioada studiului au fost nregistrate doar 5 cazuri de
PDNS. Animalele au fost examinate clinic, dup care au fost eutanasiate pentru
examenul anatomopatologic i au fost recoltate probe pentru examenul
histopatologic i pentru detectarea circovirusului porcin tip 2 (PCV2) din esuturile
afectate.

REZULTATE I DISCUII

Tabloul clinic. Manifestrile clinice pe care le-am observat au fost similare


cu cele descrise n bibliografia internaional. Suinele afectate au fost cu vrste
cuprinse ntre 3-5 luni. Cele mai reprezentative semne clinice nregistrate au fost
urmtoarele: prezena unor pete hemoragice multifocale, uneori eritematoase, de
dimensiuni cuprinse ntre 1-4 cm, uneori mai mari, cu confluarea acestora
prezente pe ambele pri ale corpului, mai ales la nivelul membrelor posterioare,
regiunea perineal, zona flancului, regiunea inferioar a abdomenului. La 4 din
cele 5 cazuri de PDNS, leziunile cutanate au fost prezente si la nivelul membrelor
anterioare, respectiv la nivelul urechilor (Fig. 1-2). Aceste animale au fost
anorexice, apatice, cu edeme declive mai ales la nivelul membrelor posterioare.

1 2
Fig.1, Fig. 2. Aspectul clinic al suinelor afectate de PDNS. Zone ntinse de pete
hemoragice pe membre, zona inferioar a abdomenului i a urechilor

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Tablou anatomopatologic. Leziunile majore semnificative au fost


nregistrate la nivelul pielii, rinichilor i limfoganglionilor. Leziunile cutanate au
fost cele descrise la tabloul clinic, cu meniunea c aceste leziuni au fost prezente
i subcutan (Fig. 3). S-a nregistrat o reacie limfoganglionar generalizat, mai
pronunat a celor inghinalor superficiali, suprarenali, submandibulari si
traheobronici. Limfoganglionii au fost foarte reacionai, congestionai, uneori
hemoragici (Fig. 4). Rinichii au fost mrii n volum, de aproximativ 3-4 ori fa
de normal, cu numeroase pete hemoragice att pe suprafa ct i n profunzimea
organului (Fig.5). Din punct de vedere histologic principalele modificri au
fost :la nivelul pielii, vasculite necrotice severe generalizate; de la nivelul
rinichilor, glomerulonefrite fibrinonecrotice, supurate, necroza fibrinoid a
vaselor de snge, trombi hialini n tubii uriniferi, infiltrat masiv limfocitar
perivascular, corpusculi renali modificai cu acumulare de material proteic n
spaiul Bowmann, uneori chiar hematii, pe alocuri cu aspectul vestigial al
corpusculilor renali (Fig. 7); de la nivelul limfoganglionilor, limfadenit
hemoragico-necrotic cu tendin de omogenizare a foliculilor, dispariia
foliculilor secundari germinativi, uneori cu apoptoz limfocitar masiv.

Fig. 3. Aspectul leziunilor cutanate la un


Fig. 4 Limfoganglionii de la un caz de PDNS.
caz de PDNS.

Fig.6. Rinichi de la un caz de


Fig 5. Rinichi de la un caz de PDNS, mrit
PDNS.Glomerulonefrita interstitila, cu
n volum cu pete hemoragice pe suprafa
afectarea severa a glomerulilor;trombi hialini
nefrit interstiial.
in tubii uriniferi.Coloratie Tricrom-Goldner.

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Diagnosticul. n ceea ce privete diagnosticul sindromului dermatitei i


nefritei porcine (PDNS) acesta se bazeaz pe urmtoarea triad: prezena
semnelor clinice caracteristice (1), prezena leziunilor histopatologice
specifice(2), i detectarea circovirusului porcin tip 2 (PCV2) din aceste leziuni.
Pentru detectarea PCV2 se utilizeaz diferite tehnici de imunodiagnostic cum ar
fi, testul ELISA, imunoperoxidaza n monostrat, imunohistochimia, i de biologie
molecular cu ar fi PCR i hibridizarea in situ. Pentru detectarea PCV2 din
esuturi am utilizat PCR (Fig. 8) i hibridizarea in situ (Fig. 7).

Fig 7. Seciune din rinichi.Detectarea ADN-ului PCV2 (culoare negricioas) n celulele


multinucleate gigante (sgeat mare) i n histiocite (sgeat mica).Hibridizare in situ.

Fig 8. Detectarea ADN-ului PCV2 de la cazurile de PDNS din limfoganglioni.


Tehnica PCR: probe pozitive (sgeata mic), martor pozitiv (cap de sgeat),
martor negativ (sgeat mare).

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CONCLUZII

Semnele clinice i leziunile anatomopatologice observate au fost


caracteristice sindromului dermatitei i nefritei porcine (PDNS) cauzat de
circovirusul porcin de tip 2, fiind corespunztoare celor descrise n literatur.
PDNS este aparent o condiie emergent caracterizat prin leziuni primare
la nivelul pielii, rinichilor i limfoganglionilor asociate cu vasculita
multisistemic. Aceste leziuni sunt probabil rezultatele reaciei de
hipersensibilizare prin complexe imune (reacie de hipersensibilitate tip III)
(Rosell i col., 2000).
PDNS a fost frecvent asociat cu PCV2 i uneori cu PRRSV, aceste virusuri
putnd fi ntlnite uneori n acelai esuturi. Rmne de determinat dac aceste
virusuri sunt factorii declanatori ai bolii, precum i modul n care ele contribuie
la dereglarea sistemului imun avnd ca rezultat acumularea patologic.
Deasemea, cunoaterea principalelor semne clinice ct i a celor
anatomopatologice n sindromul dermatitei i nefritei porcine (PDNS) este foarte
important att n direcia diagnosticului pozitiv, ct i n diagnosticul diferenial,
existnd numeroase semne clinice i anatomopatologice care se pot confunda cu
cele din alte boli, cum ar fi cele din pesta porcina clasic, rujet.

BIBLIOGRAFIE

1. Choi, C., Chae, C., 2001. - Colocalization of porcine reproductive and respiratory
syndrome virus and porcine circovirus 2 in porcine dermatitis and nephropathy
syndrome by double-labeling technique. Veterinary Pathology 38, 436441.
2. Choi, C., Kim, J., Kang, I.J., Chae, C., 2002. - Concurrent outbreak of PMWS and
PDNS in a herd of pigs in Korea. Veterinary Record 151, 484485.
3. Duran, C.O., Ramos-Vara, J.A., Render, J.A., 1997 - Porcine dermatitis and
nephropathy syndrome: a new condition to include in the di.erential diagnosis list
for skin discoloration in swine. Swine Health and Production 5, 241245.
4. Ramos-Vara, J.A., Duran, O., Render, J.A., Craft, D., 1997 - Porcine dermatitis and
nephropathy syndrome in the USA. Veterinary Record 141, 479480.
5. Rosell, C., J. Segales, J.A. Ramos-Vara, J.M. Folch, G.M. Rodriguez-Arrioja,
C.O. Duran, M. Balasch, J. Plana-Duran, M. Domingo 2000 - Identification of
porcine circovirus in tissues of pigs with porcine dermatitis and nephropathy
syndrome. Vet. Rec. 146, 4043.
6. Smith, W.J., Thomson, J.R., Done, S., 1993 - Dermatitis/nephropathy syndrome of
pigs. Veterinary Record 132, 47.
7. Thibault, S., Drolet, R., Germain, M.C., DAllaire, S., Larochelle, R., Magar, R.,
1998 - Cutaneous and systemic necrotizing vasculitis in swine. Veterinary
Pathology 35, 108116.

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TESTAREA CLINIC A UNUI VACCIN POLIVALENT


INACTIVAT DESTINAT IMUNOPROFILAXIEI UNOR
CLOSTRIDIOZE BOVINE
THE CLINICAL TRIALS OF INACTIVATED POLYVALENT VACCINE USED IN THE
SPECIFIC IMMUNOPROFILAXY OF SOME BOVINE CLOSTRIDIOSIS.

N. CTAN, V. HERMAN,
M. NECSULESCU, Ionica FODOR
U.S.A.M.V.B. Timioara

The objective of the study was to clinically test the efficiency of an


inactivated, polyvalent vaccine against some bovine clostridiosis. Twenty-
five adult, lactating cows were administered the vaccine at four weeks
interval. Local and general reactions were investigated, and the post-
vaccinal antibody titres were established using the serum neutralization
method, which was corried out on mice, for all the three antigenic
components. The antibody titres raised gradually after each vaccination, a
significant raise being registered after the second inoculation. The obtained
results recommend the use of the vaccine in bovine clostridiosis
immunoprophilaxy.

Evoluia mai multor boli toxiinfecioase, produse de bacterii din genul


Clostridium, la bovine, a determinat, n timp, obinerea unor produse
imunobiologice destinate profilaxiei acestor boli (1, 3).
Obinerea unui vaccin cu eficacitate deosebit este dependent de mai muli
factori: mediul de cultur, adjuvantul folosit, doza de inoculare i schema de
vaccinare. La aceti factori se adaug existena mai multor specii patogene
precum i a mai multor serotipuri patogene n cadrul speciei de Cl. Perfringens (2,
4, 5).
Din motivele expuse, n prezent, exist mai multe tipuri de vaccinuri
inactivate polivalente destinate profilaxiei mai multor clostridioze i care sunt
folosite la bovine i ovine (3, 4).
Cercetrile efectuate se nscriu n acest context i au avut ca obiectiv
testarea clinic, a unui nou vaccin polivalent inactivat destinat imunoprofilaxiei
unor clostridioze ale bovinelor.

MATERIAL I METOD

Vaccinul polivalent inactivat, denumit comercial ULTRACHOICE 8, a fost


testat clinic n ferma zootehnic a staiunii didactice a USAMVB Timioara, iar
examenele de laborator au fost efectuate n cadrul I.C.P.B.M.U.V. Bucureti.

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Vaccinul conine: Cl. chauvoei - tulpina F (2,28 U.O*); Cl. haemolyticum


tulpina IRP 135 (216 UT** i 3,6 U.O.); Cl. novyi tulpina 8296 (6000 D.L.M.***);
Cl. perfringens tip C tulpina PC8 (300 U.T.); Cl. perfringens tip D tulpina 317
(100 U.T.); Cl. septicum tulpina A/IRP 111 (32 U.T.); Cl. sordellii tulpina 5918
(20 U.T.); formaldehid 0,2 % i stimugen (adjuvant hidrosolubil), ph ul final fiind
de 6,2 6,4.
Masa antigenic a componentelor existente n vaccin este exprimat diferit:
U.O* (uniti de opacifiere a culturii); UT** (unitile toxinei nainte de inactivare);
D.L.M.*** (doza limit letal a toxinei nainte de inactivare).
Vaccinul a fost administrat la un lot de 25 vaci adulte aflate n lactaie (lotul
experimental), iar lotul martor (nevaccinat) a avut un efectiv egal. Vacile din
ambele loturi au fost sntoase clinic, normotermice, fiind cazate i furajate n
acelai condiii.
Vaccinul a fost administrat n doz de 2 ml s.c. n regiunea gtului, treimea
mijlocie, sub forma a dou administrri (V I a i V a II a) la nterval de 4
sptmni.
Animalele au fost examinate clinic local i general pentru depistarea
reaciilor adverse la 7, 14, 21 i 30 de zile, dup fiecare vaccinare.
Pentru testarea rspunsului imun, de la animalele vaccinate i nevaccinate,
au fost prelevate probe de snge, n vederea obinerii serurilor, dup cum
urmeaz: T0 nainte de prima vaccinare; T1 - la 4 sptmni dup vaccinare I - a
i T2 la 4 sptmni dup vaccinarea a II a.
Evaluarea rspunsului imun postvaccinal a fost efectuat prin reacia de
seroneutralizare pe oareci (5) n cadrul I.C.P.B.M.U.V. Bucureti. Serurile au fost
comasate n trei probe medii (V1, V2, V3), pentru fiecare recoltare fiind testai
numai anticorpii pentru Cl. perfringens tipul C, Cl. perfringens tipul D i Cl. novyi.

REZULTATE I DISCUII

Datele obinute furnizate n urma testrii clinice i a examenelor de


laborator au fost prelucrate i redate n tabele.
Vaccinul testat a produs, dup fiecare administrare, reacii locale exprimate
prin noduli sau edem dur a cror intensitate i durat sunt redate n tabelul 1.
Reaciile locale au fost mai intense n primele 7 zile dup inocularea vaccinului,
ulterior diminund n intensitae, astfel c la 21 zile nodulul a fost prezent la un
numr mic de animale.
Reaciile generale (febra, diminuarea produciei de lapte), au fost absente la
toate vacile vaccinate. De asemenea, nu au fost semnalate nbolnviri, la
animalele din ambele loturi, pe toat durata experimentului.
Rezultatele examenelor serologice privind rspunsul imun postvaccinal
sunt redate n tabelele 2, 3 i 4. Rspunsul imun postvaccinal a fost testat numai
pentru trei componente antigenice existente n structura vaccinului. Rspunsul
imun postvaccinal, respectiv titrul anticorpilor obinui prin reacia de
seroneutralizare pe oareci a fost exprimat n U.I/ ml.

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Tabelul 1
Reaciile locale i evoluia lor la bovinele vaccinate cu vaccinul ULTRACHOICE 8

7 zile 14 zile 21 zile Vacci- 7 zile 14 zile 21 zile


Nr. Nr. Vaccinarea
dup dup dup narea dup dup dup
crt. matricol ntia
V1 V1 V1 a doua V2 V2 V2
0 1 2 3 4 5 6 7 8 9
1 784 V1 ED=10 N - V2 - - -
2 776 V1 ED=10 N - V2 ED=5 N -
3 735 V1 ED=10 ED=5 N V2 ED=10 N N
4 813 V1 ED=10 N - V2 ED=5 - -
5 547 V1 ED=10 ED=5 N V2 N - -
6 543 V1 ED=10 N - V2 ED=10 N -
7 515 V1 N N - V2 N - -
8 506 V1 N - - V2 ED=5 - -
9 1051 V1 ED=5 - - V2 ED=10 N -
10 1074 V1 N - - V2 ED=5 - -
11 1000 V1 N - - V2 N - -
12 787 V1 ED=5 N - V2 ED=10 N -
13 799 V1 ED=10 N - V2 N - -
14 504 V1 ED=10 N N V2 ED=10 N -
15 1080 V1 ED=10 - - V2 ED=5 - -
16 801 V1 N - - V2 - - -
17 986 V1 N - - V2 N - -
18 606 V1 - - - V2 - - -
19 717 V1 ED=10 N - V2 N - -
20 514 V1 ED=5 - - V2 ED=5 N -
21 508 V1 ED=10 N - V2 ED=5 - -
22 749 V1 ED=5 N - V2 N - -
23 650 V1 N - - V2 ED=5 N -
24 741 V1 - - - V2 N - -
25 643 V1 N N - V2 N - -
Legend:
- ED=10 i ED=5 reprezint reacii locale sub forma unor edeme dure cu diametrul
de aproximativ 10 sau 5 cm;
- N = nodul (diametrul 2 cm).

Analiznd rezultatele obinute se obser c nainte de vaccinare animalele


nu au avut anticorpi fa de cele trei componente antigenice testate, valorile
obinute fiind egale cu limitele de admisibilitate stabilite de firma productoare.
Dup vaccinarea Ia valorile rspunsului imun au depit limitele de
admisibilitate pentru cele trei componente antigenice, valorile probelor medii
exprimate n U.I/ml fiind prezentate n tabele 2, 3 i 4.
Dup vaccinarea a II a (de rapel) valorile rspunsului imun postvaccinal
au fost foarte semnificative, mediile geometrice fiind net superioare limitelor de
admisibilitate pentru toate cele trei componente antigenice testate (tabele 2, 3 i
4).

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Tabelul 2
Rezultatele examenelor serologice la vacile imunizate cu vaccinul ULTRACHOICE 8
(Clostridium perfringens tipul C)

admisibilitate

admisibilitate

admisibilitate
comasat

Limita de

Limita de

Limita de
Valoarea

Valoarea

Valoarea
obinut

obinut

obinut
matricol
Numr

Proba

T0

T0

T1

T1

T2

T2
784
776
735
813
547 0,1 <0,1 >6 10
V1 3 U.I./ml 20 U.I./ml
543 U.I./ml U.I./ml U.I./ml U.I./ml
515
506
1051
1074
1000
787
799
504
1080 0,1 <0,1 >6 10
V2 3 U.I./ml 15 U.I./ml
801 U.I./ml U.I./ml U.I./ml U.I./ml
986
606
717
514
508
749
0,1 <0,1 >6 10
650 V3 3 U.I./ml 15 U.I./ml
U.I./ml U.I./ml U.I./ml U.I./ml
741
643

V1+V2+V3= 16,5 U.I./ml (MG)

Intensitatea rspunsului imun postvaccinal este consecina utilizrii, n


compoziia vaccinului, a unor tulpini bine selecionate, bune productoare de
toxin, dar i a utilizrii unui adjuvant modern, care stimuleaz rspunsul imun
postvaccinal (4, 5).

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Tabelul 3
Rezultatele examenelor serologice la vacile imunizate cu vaccinul ULTRACHOICE 8
(Clostridium perfringens tipul D)

admisibilitate

admisibilitate

admisibilitate
comasat

Limita de

Limita de

Limita de
Valoarea

Valoarea

Valoarea
obinut

obinut

obinut
matricol
Numr

Proba

T0

T0

T1

T1

T2

T2
784
776
735
813
547 <0,1 0,5
V1 0,1 U.I./ml 1 U.I./ml 2 U.I./ml 2 U.I./ml
543 U.I./ml U.I./ml
515
506
1051
1074
1000
787
799
504
1080 <0,1 0,5
V2 0,1 U.I./ml 1 U.I./ml 2 U.I./ml 2 U.I./ml
801 U.I./ml U.I./ml
986
606
717
514
508
749
<0,1 0,5
650 V3 0,1 U.I./ml 1 U.I./ml 2 U.I./ml 4 U.I./ml
U.I./ml U.I./ml
741
643

V1+V2+V3= 2,51 U.I./ml (MG)

Rezultatele obinute n urma vaccinrii, dovedesc c vaccinul


ULTRACHOICE 8 a indus un rspuns imun protectiv, cu titruri semnificative,
mai ales dup vaccinarea de rapel, ceea ce recomand folosirea acestui vaccin n
profilaxia unor clostridioze la bovine.
La lotul martor, nefiind nregistrate titruri ale anticorpilor specifici,
rezultatele nu au mai fost redate n tabele i comentate.

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Tabelul 4
Rezultatele examenelor serologice la vacile imunizate cu vaccinul ULTRACHOICE 8
(Clostridium novyi)

admisibilitate

admisibilitate

admisibilitate
comasat

Limita de

Limita de

Limita de
Valoarea

Valoarea

Valoarea
obinut

obinut

obinut
matricol
Numr

Proba

T0

T0

T1

T1

T2

T2
784
776
735
813
547 0,01 <0,01
V1 0,2 U.I./ml 3 U.I./ml 0,5 U.I./ml 6 U.I./ml
543 U.I./ml U.I./ml
515
506
1051
1074
1000
787
799
504
1080 0,01 <0,01
V2 0,2 U.I./ml 3 U.I./ml 0,5 U.I./ml 8 U.I./ml
801 U.I./ml U.I./ml
986
606
717
514
508
749
0,01 <0,01
650 V3 0,2 U.I./ml 3 U.I./ml 0,5 U.I./ml 4 U.I./ml
U.I./ml U.I./ml
741
643

V1+V2+V3= 5,76 U.I./ml (MG)

CONCLUZII

Vaccinul ULTRACHOICE 8 administrat la bovinele adulte sub forma a


dou inoculri nu a produs reacii generale, iar reaciile locale (nodul i edem dur)
s-au remis, n timp, dup fiecare inoculare;
Vaccinul ULTRACHOICE 8 a indus un rspuns imun intens numai dup
vaccinarea a doua (de rapel), diferena ntre mediile geometrice ale titrurilor
anticorpilor i limitele de admisibilitate fiind semnificativ;

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Rezultatele obinute n urma testrii clinice i de laborator recomand


utilizarea vaccinului ULTRACHOICE 8 n imunoprofilaxia unor clostridioze la
bovine.

BIBLIOGRAFIE

1. Carter, G.R., Chengapa M.M., 1993 Microbial diseases, Ed. Iowa State University
Press, Ames.
2. Gherghariu S., Pop Al., Kadar L., Marina Spnu, 2000 Manual de laborator clinic
veterinar, Ed. All Educational, Bucureti.
3. Nascimento R.A.P., Lorato F.C.F., Abreu V.L.V., Martins N.E., Assia R.A.,
Carvalho Filho M.B., 2004 Evaluation of vaccines against Clostridium novyi
type B, Brasiliero de Medicina Veterinaria e Zootehnia, 56, 1, 1-6.
4. Radostis O.M., Gay C.C., Blood D.C., Hinchcliff K.W., 2000 Veterinary Medicine
9th edition, Ed. W.B. Saunders Company Ltd., London, New York, Philadelphia,
San Francisco, St. Louis, Sydney.
5. Secaiu V., 2001 - Boli produse de germeni din genul Clostridium, n: Boli infecioase
ale animalelor - bacterioze, sub redacia Moga Mnzat Radu, pag. 481-611, Ed.
Brumar, Timioara.

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OBSERVAII PRIVIND ETIOLOGIA


PARATIFOZELOR AVIARE
Doina Anca HRU, T. PERIANU T.,
Irina-Oana TNASE, C. PAVLI
U.S.A.M.V. Iai

Considering the increasing role played by domestic and wild birds in


food poisoning in human, we started a study regarding etiology and
prevalence of paratiphic infections in birds.
121 birds were necropsied, 43 from unit A and 61 from unit B. In
unit A, 18 birds (43, 9%) been diagnosed positively, in comparison with unit
B with 22 birds (36, 06%) diagnosed positively.
The bacteriologic determinations performed from ovarian follicles,
conducted to nine Salmonella enteritidis strains and three Salmonella
typhimurium strains in comparison with liver determinations that revealed
two Salmonella enteritidis strains and one salmonella typhimurium strain.

Key words: Salmonella, food poisoning, ovarian follicles.

n trecut dintre animalele domestice, bovinele erau unul din cele mai
importante rezervoare de infecie cu Salmonella enteritidis i Salmnonella
typhimurium, n timp ce astzi, n toate rile evoluate (dezvoltate), paralel cu
extinderea creterii industriale a psrilor, acestea au devenit principalele specii
purttoare i eliminatoare. Totui, fenomenul nu se manifest stereotip
nregistrndu-se i aspecte caracteristice de la o ar la alta i chiar de la o zon la
alta.
Dintre speciile de psri, o importan sanitar veterinar mai mare o au
ginile i palmidele (raa i gsca), iar infeciile se pot grupa n dou categorii. n
prima categorie se include infecia determinat de Salmonella pulorum-
gallinarum, care afectez n special ginile i curcile determinnd tifoza i
puloroza. A doua categorie de infecie este produs de Salmonele mobile,
denumite i paratifice.
ntruct se observ o cretere a rolului psrilor domestice i slbatice, ca i
a produselor acestora (ou i carne) in apariia imbolnvirilor, dar i a
toxiinfeciilor alimentare la om, ne-am propus a investiga etiologia i prevalena
(frecvena) infeciilor paratifice la psri.

MATERIAL I METOD

Observaiile s-au efectuat n perioada 2003 - 2004, n dou uniti de


creterea psrilor pentru ou de reproducie, unde la examenul necropsic s-
au nregistrat leziuni ce suspicionau o infecie salmonelic. De menionat c

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psrile din cele dou uniti au dat n totalitate reacii negative la examenul
serologic pentru tifopuloroz.
Coninutul foliculilor ovarieni cu modificri evidente (pediculai, boselai i
de culoare verzui-albstruie) a celor102 psri, s-a nsmnat n bulion, pe
geloz nclinat sau n plci Petri i n mediul de mbogire (Kauffman-Muller).

REZULTATE I DISCUII

S-au necropsiat un numr de 102 psri, din care 43 din unitatea A i 61 din
unitatea B. Din unitatea A au fost diagnosticate cu leziuni de paratifoz 18
(43,9%) psri, iar din unitatea B 22 de psri, adic 36,06%, (tabel 1, fig. 1)

Tabelul 1
Rezultatul examenului necropsic

Nr. Leziuni de paratifoz


Unitatea Psri necropsiate
crt. Nr. %
1. A 41 18 43,9%
2. B 61 22 36,06%
Total 102 40 39,2%

70
60
50
40 A
30 B
20
10
0
P s ri n e c ro p s ia te N r. L e z iu n i d e
p a ra tifo z

Fig. 1 Reprezentarea examenului necropsic

Leziunile anatomopatologice sunt redate n tabel 2 i fig. 2.

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Tabelul 2
Leziunile anatomopatologice

Leziuni macroscopice
Nr. Nr. psri Ficat
Unitatea Peritonit Spleno-
crt. necropsiate Salpingite Ovarite
vitelin
mrit
megalie
hipercolie
Nr. % Nr. % Nr. % Nr. % Nr. %
1. A 41 6 14,65 14 36,58 7 14,87 6 14,42 8 19,84
2. B 61 9 14,75 16 26,95 15 23,88 11 18,03 10 16,39
Total 102 15 14,70 30 29,70 22 21,56 17 16,66 18 17,38

35
30
25
20 n r.
15 %
10
5
0
S a lp in g ite O v a r ite P e rito n it S p le n o - F ic a t m rit
v ite lin m e g a lie h ip e rc o lie

Fig. 2 Leziunile anatomopatologice

S-au observat un nr. de 15 (14,70%) leziuni de salpingit, 30 (29,70%)


leziuni de ovarit, 22 (21,56%) leziuni de peritonit vitelin, 17 (16,66%) cazuri
de splenomegalie i 18(17,38%) cadavre cu ficat mrit.
Pentru izolarea tulpinilor de salmonela i confirmarea suspiciuniide infecie
paratific, nsmnrile pentru examenul bacteriologic s-au efectuat din foliculii
ovarieni modificai i din ficat. n acest scop s-au folosit medii de cultur
obinuite (geloz i bulion), medii de mbogire (Kauffman-Mler) i medii
selective (Wilson-Blair).
Coloniile de pe mediile selective cu aspect asemntor celor de salmonela
au fost examinate n ceea ce privete aglutinabilitatea cu ser antisalmonelic
polivalent O (A, B, C, D, D ). Cele pozitive au fost transplantate pe geloz
nclinat i cercetate cu privire la aspectul cultural, aglutinabilitatea cu ser de
grup, mobilitatea i caracterele biochimice.
Pe geloza nclinat unele tulpini s-au dezvoltat sub form de colonii mari,
rotunde, cu marginile netede asemntoare celor de colibacili, iar altele sub form

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de colonii mici asemntoare celor de pasteurela. Toate tulpinile izolate au fost


mobile. Rezultatele examenului bacteriologic sunt redate n tabel 3 i fig. 3.
Tabelul 3
Rezultatele examenului bacteriologic

Nr.
Salmonella Salmonella Esch. Germeni Proteus
Nr. tulpini
Organ enteritidis typhimurium coli coliformi v.
probe izolate
Nr. % Nr. % Nr. % Nr. % Nr. % Nr: %
Ovar 40 21 52,5 9 42,85 3 14,29 4 19,05 4 19,05 2 9,52
Ficat 40 22 55,0 2 9,07 1 4,58 8 4,58 8 36,36 4 18,18

25

20

15
numar

ovar
ficat
10

0
Nr. tulpini Salmonella Salmonella Esch.coli Germeni Proteus v.
izolate enteritidis typhimurium coliformi

Fig. 3 - Rezultatele examenului bacteriologic

Din rezultatele obinute se constat c germenii izolai din ficat i din ovar
difer ca pondere.Astfel nsmnrile din foliculii ovarieni au dus la izolarea a 9
(42,85%) tulpini de Salmonela enteritidis i a 3 (14,29%) tulpini de Salmonela
typhimurium, n timp ce din ficat s-au izolat numai 2 (9,07%) tulpini de
Salmonela enteritidis i 1 (4,58%) tulpini de Salmonela typhimurium. n schimb
n ficat au predominat ali germeni Gram negativi cum sunt Escherichia coli 8
(36,36%) tulpini,germeni coliformi 7 (31,81%) tulpini i 4 (18,18%) Proteus
vulgaris. De menionat c de la 3 psri din ovar s-au izolat att Salmonela
enteritidis ct i Escherichia coli.
Datele din literatura de specialitate redau c n Romnia, incidena
infeciilor cu salmonele mobile la psri investigat ntr-un centru de selecie i
hibridare a raselor grele i unul de hibridare al raselor uoare a fost: Salmonella
enteritidis 50,8% , Salmonella typhimurium 30,4%, Salmonella gallinarum 5,8%,
Salmonella heidelberg 2,4% , Salmonella agona 2,10% , Salmonella derby 2,9% ,
Salmonella haifa 2,4% , Salmonella newport 2,4%.

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Rezultatele obinute evideniaz faptul c din foliculii ovarieni modificai


se pot izola salmonele mobile,n special Salmonela enteritidis i Salmonela
typhimurium. Aceste date sunt asemntoare cu cele obinute de Robert Davies i
col.2oo3 care arat c salmonelele mobile se pot izola din foliculii ovarieni dar nu
se cunoate dac acestea se pot transmite la patogeni i ce rol epidemiologic joac
n difuziunea infeciei.

CONCLUZII

Investigaiile efectuate au dus la desprinderea urmtoarelor concluzii:


1. Examenul necropsic a 102 psri a evideniat leziuni de paratifoz n
39,20% cazuri.
2. Leziunile evideniate au fost de salpingit (14,70%), ovarit (21,56%),
peritonit vitelin (16,66%) i spenomegalie (17,38%).
3. nsmnrile efectuate din ficat i ovar pe medii de cultur, au dus la
izolare din foliculii ovarieni a 9(42,85%) tulpini de Salmonela enteritidis i 3
(14,29%) tulpini de Salmonela typhimurium, iar din ficat a 2 (9,07%) tulpini de
Salmonela enteritidis i 1 (4,58%) tulpini de Salmonela typhimurium.

BIBLIOGRAFIE

1. Carp-Crare M., Guguianu Eleonora, Timofte Dorina, 1997 Lucrri practice de


Microbiologie veterinar Uz intern, Universitatea Agronomic i de Medicin
Veterinar Ion Ionescu de la Brad Iai.
2. Cooper & Gerard I., 1994 Salmonellosis infections in man end the chicken
pathogenesis and the development of live vaccines a review Veter. Bull.
Vol.64, no.2,123-135.
3. Davies R., Liebane E., Breslin M., 2003 - Investigation of the distribution and control
of Salmonella enterica serovar Enteritidis PT6 in layer breeding and egg
production. Avian Pathology (June 2003) 32(3),227-237.
4. Gast R.K., Holt P.S., 1998 - Persistence of Salmonella enteritidis from one day of
age until maturity in experimentaly infected layer chickens.-Poultry Science, 77
m1 759-1762.
5. Keller L.H., Schifferli D.M. ,Benson C.E. , Aslam S., Eckroade R.J., 1997 -
Invasion of chicken reproductive tissues and forming eggs is not unioue to
Salmonella enteritidis. Avian Diseases , 41, 535-539.
6. Mnzat Moga R., 2001 - Bolile infecioase ale animalelor bacterioze Ed. Brumer,
Timioara.
7. Perianu T. (coordonator), 2003 - Bolile infecioase ale animalelor domestice.
Bacterioze. Vol l, Ed. Venus, Iai.

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DETERMINAREA PATOGENITII SPECIILOR


BACTERIENE IMPLICATE N PATOLOGIA INFECIOAS
DE INCUBAIE
EGGS INCUBATION PATHOLOGY AND PATHOGENICITY
OF THE BACTERIA ISOLATED

Emilia CIOCRLAN, T. PERIANU,


Irina-Oana TNASE, C. PAVLI
U.S.A.M.V. Iai

The isolated germs were tested regarding pathogenicity in vivo and


in vitro.
Tests were conducted on mousses using 0, 1-0,4ml intraperitoneal
inoculation.
The isolated and analyzed strains proved to be pathogen inducing
death to test animals in eighteen twenty-four hours.

Unele indicii privind patogenitatea bacteriilor pot fi obinute in vitro pe


baza testelor biochimice, n cazul cnd unele activiti enzimatice sunt corelate cu
intervenia lor ca factori de agresivitate. Aa este cazul testului de hemoliz, de
coagulare a plasmei citrate la stafilococi etc.
Tehnicile in vitro dau ns indicaii mult mai precise asupra patogenitii
germenilor.

MATERIAL I METOD

Patogenitatea in vitro s- a analizat pe baza caracterelor biochimice.


La Staphylococus aureus s-a studiat activitatea hemolitic pe geloz cu 5%
snge de oaie defibrinat.
Activitatea cuagulazic s-a efectuat n tub care evideniaz coagularea
liber.
Activitatea fosfotazic s-a studiat pe medii care conin fenolftalein difosfat.
Patogenitatea in vivo a fost cercetat prin inoculri la animale de laborator, pe
oarece fiind executate inoculri cu doze subletale, n puncte separate, pe cale
subcutan.

REZULTATE I DISCUII

Hemolizina a produs o zon de hemoliz total, iar hemolizina o zon de


hemoliz parial, dar care s-a identificat devenind total n situaia cnd culturile
au fost trecute de la 37oC la 4oC.

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Activitatea cuagulazic s-a determinat amestecnd 0,9 ml suspensie de


germeni cu 0,1 ml plasm recoltat EDTA. S- a incubat amestecului la 37oC i s-a
urmrit formarea coagului n interval de 30 minute 3 ore.
S-a mai recurs la testul de aglutinare pe lam, mai rapid obinndu-se
apariia coagului n 10 secunde i mai economic, suspensia de germeni se
amestec cu o pictur de plasm.
Mediile care conin fenolftalein difosfat sunt descompuse de fosfataz cu
eliberare de fenolftalein a crei prezen s-a pus n eviden prin adugarea la
cultur a 0,5 ml soluie aminoantipirin (6%) i soluie 2,4% de ferocianur de
potasiu. Culoarea rou intens a indicat activitatea fosfatazic.
Caracteristicile biochimice ale genurilor familiei Enterobacteriaceae
studiate sunt prezentate n tabel 1.
Tabelul 1
Caracterele biochimice difereniale ntre principalele genuri
ale familiei Enterobacteriaceae

Escherichia Salmonella Proteus


Mobilitate + + +
Indol + - d
H2S - d d
Ureaz - - +
Citrat - d d
Malonat - d -
Mucat + d -
KCN - d +
Fenilalanindeaminaza - - +
Lizindecarbozilaza + + -
Omitindecarboxilaza d d d
Argininhihrolaza d d -
Lichefierea gelatinei - - +
Rou metil - + +
Voges-Proskauer - - d
D-Glucoza fr gaz + + +
D-Glucoza cu gaz + d +
Lactoza d - -
Betagalactozidaza + - -
Zaharoza d - d
Maltoza + + d
Trehaloza + d +
Dulcitol d d -
D-manitol + + -
D-adonitol - - d

Patogenitatea in vivo a fost studiat pe oarecii de laborator, dozele


inoculate fiind reprezentate n tabelul 2.

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Tabelul 2
Dozele germenilor inoculai

Specia Proteus Staphylococus Escherichia Salmonella


Cantitate cultur inoculat ml 0,1 0,4 0,1 0,2
Cantitate cultur inoculat ml 0,2 0,3 0,2 0,3
Cantitate cultur inoculat ml 0,3 0,2 0,3 0,4

oarecii inoculai au fost omori n 18 24 ore, timp sensibil mai redus


dect al loturilor martori, inoculate de asemenea cu doze subletale, la care
moartea s-a produs n 7 pn la 13 zile. La nivelul locului de inoculare s-a constat
prezena unui edem mai mult sau mai puin ntins. De la oarecii inoculai s-au
reizolat n toate cazurile tulpinile cu care s-a efectuat infecia experimental.
oarecii inoculai au fost omori n 14-20 ore, timp sensibil mai redus
dect al loturilor martori, inoculate de asemenea cu doze subletale, la care
moartea s-a produs n 5 pn la 12 zile. La nivelul locului de inoculare cu Proteus
s-a constat prezena unui edem mai mult sau mai puin ntins. De la oarecii
inoculai s-au reizolat n toate cazurile tulpinile cu care s-a efectuat infecia
experimental.

CONCLUZII

Investigaiile au dus la sintetizarea urmtoarelor concluzii:


1. Infecia experimental pe oarece prin asocierea tulpinilor Proteus
vulgaris Escherichia coli n doze subletale, indic potenarea reciproc a
acestora.

BIBLIOGRAFIE

1. Brzoi D., Meica S., Negu M., 1999 Toxiinfeciile alimentare. Editura Diacon
Coresi, Bucureti.
2. Bergdoll M.S., 1998 Staphylococcus aureus in: Foodborne Bacterial Pathogenes.
Ed. By M.P.Doyle (Marcel Dekker Inc., New York, Basel).
3. Cooper &Gerard I., 1994 Salmonellosis infections in man end the chicken
pathogenesis and the development of live vaccines a review Veter. Bull.
Vol.64, no.2,123-135.
4. Carp Crare M., Guguianu Eleonora, Timofte Dorina, 1997 Lucrri practice de
microbiologie veterinar. U.S.A.M.V. Iai.
5. Davies R., Liebane E., Breslin M., 2003 - Investigation of the distribution and control
of Salmonella enterica serovar Enteritidis PT6 in layer breeding and egg
production. Avian Pathology (June 2003) 32(3),227-237.
6. Mnzat Moga R., 2001 -Bolile infecioase ale animalelor bacterioze Ed. Brumer,
Timioara.
7. Perianu T., (coordonator), (2003) - Bolile infecioase ale animalelor domestice.
Bacterioze. Vol l, Ed. Venus, Iai.

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CTEVA ASPECTE VIRUSOLOGICE N GRIPA AVIAR


SOME VIRUSOLOGICAL ASPECTS IN BIRD FLU

Elena IAN, tefania MERTICARIU,


Iulia FLORITEAN, Eugenia BURTAN
L.S.V.S. Iai

The name of bird flu or avian influenza includes several


pathological states, produced by various flu virus sub-types.
The flu virus forms part of the Influenzavirus group, A type (species).
The A flu virus infects both human beings and some animals (such as
horses, pigs, birds) and is responsible for the emergence of the pandemics
and extended epidemics.
All fowl and wild birds are considered to be predisposed to bird flu
infection. Birds of passage (wild ducks especially) are the natural cause for
bird flu, these birds being at the same time the most resistant to this
infection. The outbursts of the most pathogenic forms up to now have been
caused by the A-type flu virus (the H5 and H7 sub-types). The H5N1 flu
represents a threat for the public health, due to the emergence of the bird
flu cases in human beings and the danger of occurring a new pandemic.
Currently, efforts are being made to eliminate the virus reservoir, to
monitor the flu virus isolates, to implement new efficient measures that will
considerably limit the virus transmission to human beings, and to develop
an efficient vaccine against the avian H5N1 strains.
A number of tests were made in the county of Iasi, and the laboratory
exams for the diagnosis of the bird flu were made using 3 diagnosis
methods: 874 tests made using the immunochromatographic test or the
rapid immunomigration on cloak pads, 1480 tests were made using the
ELISA test on serum samples from birds and 1109 tests, using the PCR test
on cloak pads and trachea pads and the organic tests, which made us
conclude that the county of Iasi is free of this horrible disease.
From a total of 3463 laboratory exams, a dead turkey hen,
originating from Belcesti, proved to be positive both to tic and PCR.

Key words: H5N1 avian flu virus, pandemic, monitoring, prevention, Iasi,
immunochromatographic kit, PCR.

Denumirea de grip sau influen aviar include mai multe stri patologice,
produse de ctre diverse subtipuri ale virusurilor gripale. Cea mai cunoscut
form de grip aviar este cea numit pest aviar clasic, care nu a mai aprut n
ara noastr din anul 1941. n ultimii ani etiologia gripei a fost intens studiat i
este bine cunoscut, constituind un model de adaptare a virusului la diferite gazde,
cu consecine importante sub aspect epidemiologic. Virusul gripal este ncadrat n
grupul Influenzavirus, tipul (specia) A. Virusul gripal A infecteaz att omul ct i

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

unele animale (calul, porcul, psrile) i este responsabil de apariia pandemiilor


i a epidemiilor extinse. Acesta are dimensiunea de aproximativ 80-120 nanometri
i prezint n zona central un filament de ARN. Este un virus relativ mare i
conine 8 bucle de ARN, ce faciliteaz reasortarea (deci modificarea
structural a urmtoarelor generatii de virioni), atunci cnd aceeai celul este
infectat n acelai timp cu dou virusuri gripale.
nveliul extern al particulei virale - numit pericapsid, este de natur
lipoproteic i prezint pe suprafa dou tipuri de spiculi, alctuii din
hemaglutinin (H) i neuraminidaz (N). Hemaglutinina (H) joac un rol
important n adsorbia virusurilor pe suprafaa celulei - gazd i n realizarea
infeciei virale, iar neuraminidaza (N) este o enzim care determin eliberarea
virusului din celule i confer capacitatea de a se rspndi repede.
Sub aspect epidemiologic, virusurile gripale (de tip A) sunt submprite n
subtipuri pe baza caracteristicilor antigenice ale glicoproteinelor de suprafa (H
i N). Se cunoate existena a 16 subtipuri distincte dup hemaglutinin (H) i 9
subtipuri dup neuraminidaz (N). Aceste antigene sunt supuse variaiei.
Subtipurile se caracterizeaz fiecare printr-o formul antigenic proprie, care
reflect asemnrile i deosebirile dintre diferitele tulpini.
Gripa aviar, o boal infecioas a psrilor cauzat de mutaia de tip A a
virusului gripal a fost descoperit n urm cu mai mult de 100 de ani n Italia, de
atunci rspndindu-se n ntreaga lume. Toate psrile domestice i slbatice, sunt
considerate predispuse la infecie cu gripa aviar (palmipedele, gina, curca, mai
rar fazanul, punul, bibilica, sunt mai sensibile psrile adulte dect puii), dei
unele specii sunt mai rezistente la infecie dect altele. (Doar un amunit izolat
viral poate produce o boal sever la galinacee, fr ca alte specii de pasri s
fie afectate clinic).
Psrile migratoare (raele slbatice, n mod special) sunt sursa natural de
grip aviar, aceste psri fiind de asemenea cele mai rezistente la infecie.
Psrile domestice n schimb, inclusiv ginile i curciile sunt deosebit de
predispuse la epidemiile de grip fatale. Evoluia la palmipede poate fi mai puin
sever, iar n cazul raelor n multe cazuri chiar inaparent, dar acestea rmn
purttoare i excretoare de virus. Infecia a fost descris la foarte multe specii de
psri slbatice, mai frecvent la cele acvatice, cu un procent de mortalitate
foarte variabil (0-90%). Infecia a fost descris i la multe specii de mamifere, la
care s-au observat forme severe de boal, de exemplu la pisici slbatice i
domestice, porc i anima1e n captivitate, (tigri, leopard, nevstuic, maimu,)
reuindu-se chiar infecie experimental pe pisici domestice, iepuri albi
neozeelandezi i obolan.
Omul face de asemenea forme foarte severe de boal cu procent mare de
mortalitate, consecutiv contactului nemijlocit cu psri bolnave sau moarte.
Izbucnirile formelor foarte patogenice pn acum au fost cauzate de virusul
gripal de tip A (subtipurile H5 si H7).

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Universitatea de tiine Agricole i Medicin Veterinar Iai

Prima infecie cu un virus de grip aviar nregistrat la om a aprut n


Hong Kong n 1997, cnd tulpina de tip H5N1 a cauzat boli respiratorii acute la
18 persoane, dintre care 6 au murit. Infecia la oameni a coincis cu o epidemie de
gripa aviar, cauzat de aceeai tulpin, la populaia de psri domestice din Hong
Kong. Cea mai grava pandemie de gripa nregistrat n istorie a avut loc n 1918 i
a ucis 40 de milioane de oameni.
n continuare vom prezenta unele caracteristicii privind semnele clinice ale
acestei bolii devastatoare.
Perioada de incubaie este obinuit de 3-7 zile, fiind dependent de
patogenitatea inocului de virus, de doza de inocul, de specie i de vrsta psrilor.
Boala evolueaz n mod normal supraacut, cu moarte subit a psrilor afectate n
24 sau 48 ore, n unele cazuri poate evolua acut, cu o durat de cel mult o sptmn.
Manifestrile clinice sunt urmarea tulburrilor circulatorii, respiratorii
digestive, reproductive i a sistemului nervos: moarte subit fr manifestri
clinice; adinamie i depresie puternic asociat cu horiplumaie; inapeten,
asociat adesea cu sete excesiv; scderea dramatic a produciei de ou,
ntlnindu-se adesea ou cu coaja moale sau cu defecte de structurare i de
form a cojii; edemele crestei, brbielor i a zonei perioculare. Uneori edemul
capului este extins i n zona gtului mai frecvent la ginile adulte; crestele sunt
adesea cianotice la vrf i pot avea mici vezicule cu plasm sau chiar snge pe
suprafa uneori asociate cu hemoragii echimotice sau focare de necroz;
colorare purpurie sau cianotic a crestei, brbielor i uneori a picioarelor;
mucoasa conjunctiv este congestionat sau edemaiat; pe pielea solzoas a
picioarelor, se pot ntlni hemoragii difuze; diareea iniial apoas, verzuie poate
deveni pe parcursul bolii aproape n totalitate alb; manifestrile respiratorii,
constnd din jetaj i acumulri de mucoziti adesea sanguinolente, dispnee i
respiraie zgomotoas, pot fi de asemenea un semnal timpuriu al bolii;
manifestrile nervoase constau n torticolis, ataxie i lips de coordonare a
micrilor.
Modificrile anatomopatologice constatate la psrile moarte n urma
unor forme supraacute de boala i psrile tinere nu sunt reprezentate de leziuni
importante, doar congestia sever a musculaturii i deshidratare.
Evoluiile mai lungi sau n cazul ginilor adulte determin urmtorul tablou
lezional:
- edeme subcutanate ale capului i gtului;
- vasele de snge sunt puternic ngroate;
- acumulri de lichide n nri i cavitatea bucal;
- mucoasa conjunctival congestionat;
- coninut abundent de exsudat n trahee sau traheit hemoragic;
- hemoragii peteiale pe suprafaa grsimii abdominale i a seroaselor,
n masa musculaturii pectorale, pe suprafaa cordului, n proventricul,
pe mucoasa i seroasa intestinului;

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- rinichii puternic congestionai;


- ovare hemoragice sau degenerate, n cavitatea peritoneal se pot gsi
pseudoconcremente de ou.

MATERIAL I METOD

Studiul a fost realizat pe un numr de 539 probe recoltate de la psri n


perioada noiembrie decembrie 2005, dup cum urmez:
Tabelul 1

Specificaie probe
Nr.
Specia Tampoane
Crt. Seruri
cloacale
1 Galinacee 38 401
2 Palmipede 4 91
3 Psri slbatice 5 -
Total 47 492

Iar n perioada 01.01. 25.05.2006, cercetrile au fost efectuate pe un


numr total de 2 922 probe, astfel:
Tabelul 2

Specificaie probe
Nr.
Specia Tampoane
crt. Seruri
cloacale
1 Galinacee 219 188
2 Palmipede 13 800
3 Psri slbatice 107 -
345 -
4 Psri vii - galinacee i palmipede
141 -
TOTAL 825 988

Din totalul de 3 463 teste de laborator efectuate, 1 109 probe (organe i


tampoane cloacale) sunt reprezentate de analize efectuate prin reacia
polimerazei n lan (PCR).
Ca metode de lucru au fost utilizate: examenele clinice pe psrii vii,
examen necropsic (leziunile ntlnite la psri cu influen nu pot fi difereniate de
leziunile ce se gsesc n forma velogenic viscerotrop a bolii de Newcastle);
efectuarea testelor rapide pentru detecie de antigen al virusului influenei aviare
pe probele de tampoane cloacale i teste ELISA pe probe de ser.
Ca teste rapide de detecie a antigenului virusului influenei aviare din
tampoane cloacale sau fecale s-a utilizat:
- kitulul imunocromatografic Anigen Rapid AIV Ag Test Kit i a variantei
acestuia H5 AIV Ag Test Kit, produse de Animal Genetics Inc. din Corea, care are
ca principiu: detecia calitativ a antigenului viral al influenei A, sau a antigenului
viral al influenei A tipul H5. n dispozitivul nchis de testare (cuv) exist 2 litere

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"T" i "C" n dreptul crora vor deveni vizibile dup efectuarea testului linii
purpuriu-grena. Aceste linii nu exist iniial i definesc validarea testului (n
dreptul literei C) indiferent dac proba este pozitiv sau negativ i prob pozitiv
(atunci cnd apare n dreptul literei T). Testul utilizeaz un panel de anticorpi
selectat n scopul deteciei virusului IA.
- Trusa Flu DetectTMTest Strip este un test in-vitro de imunomigrare rapid
de detecie calitativ a virusului influenei tip A n tampoanele traheale i cloacale
de pasre. Acest test detecteaz cele 15 subtipuri ale virusului influenei tip A.
Testul utilizeaz doi anticorpi specifici virusului influenei tip A (un anticorp anti-
influena A leag antoigenul present n prob i un al doilea anticorp de captur
pentru acest complex atg-atc.).
Iar pe probele de seruri de pasre s-au efectuat teste ELISA:
- Detecia rapid de anticorpi ai influenei aviare, pe seruri de la gini i
curci vii, prin indirect ELISA (kitulul "FlockChek", produs de firma IDEXX
Laboratories din SUA. Testul msoar nivelul relativ al anticorpilor produi ca
urmare a contactului nemijlocit cu virus al influenei aviare. Pe microplcile
cptuite cu antigen viral sunt repartizate i incubate seruri. n cazul serurilor
pozitive, anticorpii: anti virus a'l influenei aviare, vor forma imunocomplexe cu
antigenul de cptuire a micrciplcii, aceste complexe fiind ulterior vizualizate
dup legarea cu un conjugat anti IgG de gin i curc, marcat cu peroxidaz,
prin adugarea unui substrat TMB, ce va produce n prezena peroxidazei o
culoare albastr cu intensitate dependent de nivelul relativ al anticorpilor anti
virus al influenei aviare existent n probe.
- Detecia rapid de anticorpi ai influenei aviare, pe seruri de palmipede,
prin ELISA competitive.

REZULTATE I DISCUII

n urma efecturii examenelor de laborator n direcia diagnosticrii gripei


aviare pe un numr de 3463 de probe, prin cele 3 metode de diagnostic:
- Testul imunocromatografic rapid (t.i.c.) sau imunomigrare rapid pe
tampoane cloacale (874 probe) recoltate de la psri vii i cadavre;
- Testul ELISA pe probe de ser provenite de la pasri vii (1480 probe);
- Reacia polimerazei n lan (PCR) efectuat pe tampoane cloacale i
traheale i probe organe, recoltate de cadavre de psri domestice i slbatice.
Recoltarea probelor de seruri i a tampoanelor cloacale de la pasri vii, a
vizat n special localitile int, n numr de 32 la nivelul judeului Iai, stabilite
pe baza riscului pe care l-au prezentat prin existena suprafeelor de luciu de ap i
a psrilor slbatice.
Au fost monitorizate toate localitile int prin t.i.c., frecvena probelor
recoltate a respectat prevederile Ord. 148/2006, de 20 tampoane/localitate int.
De asemenea, au fost efectuate t.i.c. la toate cadavrele de psri sosite la nivelul
laboratorului.
Teste ELISA s-au efectuat pe probe de ser recoltate de la psri domestice
vii, conform prevederilor legale.

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Au fost efectuate analize PCR la cadavrele de psri domestice i slbatice


sosite la nivelul laboratorului sanitar veterinar i care indicau unele modificri
morfopatologice similare celor care se ntlnesc n gripa aviar. Rezultatele
obinute au condus la concluzia c n judeul Iai s-au luat msuri eficiente de
supraveghere a teritoriului pentru aceast epidemie.
n urma efecturii testului imunocromatografic pe cele 874 probe de
tampoane cloacale recoltate de la: 602 galinacee, 158 palmipede i 112 psri
slbatice, rezultatul testelor a fost negativ.
Rezultatul unui test rapid imunocromatografic efectuat pe tampoane
cloacale, pentru detecia de antigen viral al virusului influenei aviare tip A, a fost
pozitiv (Fig. 1). Acest test pozitiv a reprezentat un cadavru de curc care provenea
din GP, localitatea Belceti. Investigaiile au fost continuate cu efectuarea de teste
PCR, att pe tampoane cloacale, traheale, ct i din organele interne i creierul
recoltate de la cadavrul de curc suspect de A150. Rezultatele obinute au fost
pozitive, motiv pentru care s-au expediat probe de tampoane cloacale, traheale i
omogenate de organe la IDSA Bucureti pentru confirmare.
n localitatea Belceti s-au efectuat teste rapide la toate gospodriile
nvecinate i prin sondaj la ntreg teritoriul comunei Belceti, dar nu s-a mai
obinut nici o reacie pozitiv, prin testul rapid imunocromatografic.
La nivelul laboratorului, din totalul de 825 de tampoane cloacale la care s-
au efectuat t.i.c., doar 3 probe (1 tampon cloacal recoltat de la o gin, 1 tampon
cloacal recoltat de la o turturic i un 1 tampon cloacal recoltat de la un porumbel
viu) au reacionat FALS pozitiv, la testul imunocromatografic de grup (acesta
poate detecta antigenul tuturor virusurilor influenei aviare de tip A de joas sau
nalt patogenitate, si nu a subtipului H5), ns la testul imunocromatografic de tip
au reacionat negativ.

Fig. 1. Test imunocromatografic cu reacie pozitiv. (curc, Belceti)

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Testul imunocromatografic de grup (fig. 2.) presupune urmtoarea


procedura de testare:

Fig.2. Test imunocromatografic

- se preleveaz un tampon cloacal; - fig.3

Fig. 3. Prelevarea probei de tampon cloacal

- se introduce tamponul n tubul cu diluant; - fig.4


- se mixeaz tamponul n diluant pn la dizolvarea coninutului; se las
puin pentru sedimentarea particulelor grosiere;
- se dezambaleaz cuva de lucru i se pune pe un suport plan i uscat;
- se extrage cu pipeta picurtor coninut de suprafa din tubul de diluie a
probei; se pun 4-5 picturi (nu mai mult), n orificiul pentru prob a cuvei de
lucru; - fig 5
- se observ culoarea roie care migreaz de-a lungul ferestrei centrale a
cuvei de lucru; fig 6.

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Fig. 5. Extragerea cu pipeta picurtor din


Fig. 4. Introducerea tamponului cloacal n
tubul de diluie a probei, a coninutului de la
tubul cu diluant
suprafa

Fig. 6. Observarea culoarei roii care Fig. 7. Prezenta unei singure benzi roii
migreaz de-a lungul ferestrei centrale a indic un rezultat negativ i valideaz
cuvei de lucru funcionarea testului

Interpretarea rezultatelor se face n 20-30 de minute i se face astfel:


- n partea dreapt a ferestrei de citire a rezultatelor va apare o band roie
ce indic funcionarea corespunztoare a testului;
- prezenta unei singure benzi roii indic un rezultat negativ; fig. 7
- prezena a dou benzi colorate n fereastra de citire (nu are important
care apare prima) indic un rezultat pozitiv;
- absenta oricrei linii colorate n fereastra de citire, indic test invalid,
fiind recomandat a se retesta aceiasi prob.
Iar n urma analizrii celor 1480 de probe ser de pasre (589 seruri
provenite de la galinacee i 891 seruri de palmipede) prin testul Elisa pentru
detecia de anticorpi, au fost obinute rezultate negative.
Prin efectuarea celor 1109 probe reprezentate de tampoane cloacale,
traheale i probe organe prin metoda PCR s-au obinut rezultate negative, cu
excepia un singur caz (un cadavru de curc) provenit din localitatea Belceti, la
care rezultatul testului PCR a fost pozitiv.

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Din extrasele de organe i tampoanele cloacale i traheale expediate la


IDSA Bucureti, s-a efectuat testul PCR, al crui rezultat a fost tot pozitiv, pe
baza acestuia a fost emis notificarea de suspiciune pentru gripa aviar pentru
judeul Iai. Notificarea suspiciunii s-a efectuat pe baza rezultatelor de laborator
coroborate cu datele furnizate de ancheta epidemiologic.
Confirmarea bolii se face numai prin izolare de virus, de ctre IDSA
Bucureti, Laboratorul Naional de Referin pentru grip aviar.
n curtea suspect identificat cu reacie pozitiv pentru A150, s-a stabilit
un complex de msuri privind prevenirea difuzrii bolii i combaterea acesteia.
Toate psrile psrile din gospodria infectat i zona stabilit de
Comandamentul Antiepizootic Judeean au fost eutanasiate.

CONCLUZII

n complexul de msuri stabilite pentru supravegherea teritoriului judeului


Iai, n direcia gripei aviare, un loc important a fost reprezentat de examenele de
laborator.
S-au efectuat teste rapide imunocromatografice pe 874 probe de tampoane
cloacale recoltate de la psri domestice i slbatice vii i cadavre ale acestora.
Teste ELISA au fost efectuate pe 1 480 probe de ser de la psri domestice i
slbatice vii
S-au efectuat teste rapide imunocromatografice de grup pentru influena
aviar tip A, pe ntreg teritoriu al judeului Iai, pentru monitorizarea virusurilor
gripale la psrile domestice i slbatice vii i de la cadavrele acestora. Detecia
timpurie a atigenului viral specific prin t.i.c. s-a continuat cu efectuarea de teste
imunocromatografice rapide de tip H5 i cu teste PCR.
Reacia polimerazei n lan s-a efectuat pe probe de tampoane cloacale i
traheale, precum i la probele de organe interne i creier provenite de la aproape
toate cadavrele psrilor domestice i slbatice expediate la laborator.
Reacia polimerazei n lan a fost efectuat pe 1 109 probe de tampoane
cloacale i traheale i organe recoltate de la cadavre de psri.
S-a identificat un caz pozitiv att la teste imunocromatografice de grup i
tip, ct i la testul PCR, pe probele recoltate de la un cadavru de curc, provenit
de la gospodriile populaiei, din comuna Belceti.
n judeul Iai s-au luat msuri concrete i eficiente de prevenire a unei
epidemii de mare difuzibilitate, de tipul gripei aviare.
Raportarea i investigarea focarelor de grip aviar are ca scop protejarea
populaiei umane.

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humans: urgent need to eliminate the animal reservoir, Jan. 2004, WHO.
9. http://www.who.int/csr/don/2005_08_18/en/index.html, Geographical spread of H5N1
avian influenza in birds, Aug. 2005, WHO.
10. http://www.who.int/csr/don/2005_05_19/en/, Avian Influenza-cumulative number of
cases, May 2005, WHO.

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EPIDEMIOLOGICAL SCREENING OVER THE


MYCOPLASMA HYOPNEUMONIAE WEARERS IN TWO
SWINE BREEDING UNITS
Cristina REBEGEA, M. CARP-CRARE,
C. CARP-CRARE
U.S.A.M.V. Iai
C. ANTON, D. FODOR
PHIZER ROMANIA SRL

Enzootic pneumoniae or mycoplasmal pneumonia of swine (MPS), a


chronic disease with a high morbidity and a low mortality, is caused by M.
hyopneumoniae. The clinical signs include a chronic nonproductive cough,
retarded growth, slow onset and spread and repeated occurence of the
disease. The DakoCytomation M. hyopneumoniae ELISA allows rapid
screening for the presence of antibodies to M. hyopneumoniae, which can
be an indicator of exposure to the agent. Monitoring the immune status of a
herd with regard to M. hyopneumoniae can play an important role in the
control of this disease.
It were tested 90 animals, four to five months old, from two breeding
units and the results showed a grately high incidence of the antibodies in
the analised serum samples (46,6%). From the first unit with imported
swine, the number of positive samples was 17 (37,7%) and from the second
unit with native swine the number of positive samples was 25 (55,5%). The
presence of the M. hyopneumoniae specific antibodies in unvaccinated
animals shows a subclinical infection or the wearer status.

Key words: ELISA, Mycoplasma hyopneumoniae, antibodies, enzootic pneumonia

M. hyopneumoniae produces swine enzootic pneumonia, a chronical


respiratory disease who afectes swines, characterized through dry cough, a
mortality rate decrease, but a very high morbidity percent. This malady major
effects consist in economical losses owened to daily weight decrease, food
conversion rate reduction, increasing of time periode till the slaughter weight is
reached, the tratament costs.
There isnt a susceptibility of age for the infection with M. hyopneumoniae,
piglets and adults being equaly affected. Piglets infect itselves through mothers
respiratory secretions after 4 - 6 weeks of age, during firsts weeks being
protected by the maternal antibodies. (Kobisch i Friis, 1996).
M. hyopneumoniae acts as a primary pathogen agent, but it can act
sinergically with other infectious agents causing a hard pneumonia, as in swine
respiratory complex case where it acts together with Porcine Respiratory and

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Reproductive Syndrome (PRSS), Pasteurella multocida, Streptococcus suis and


Actinobacillus pleuropneumoniae.
For the M. hyopneumoniae antibodies detection in swine serum it was
developed ELISA blocking (Enzyme-Linked Immunosorbent Assay) using
monoclonal antibodies. The monoclonal antibody (mAb) reacts with a conserved
epitip of the M. hyopneumoniae 74 kDa proteine. Only the antibodies from M.
hyopneumoniae infected porcine serum are capable of bloching the mAb binding,
even though M. flocculare antibodies likewise recognize a 74 kDa molecule.
ELISA blocking seems to be a helpful method for surveillance and serodiagnostic
of porcine M. hyopneumoniae infections. (Feld NC i col., 1992)
ELISA sensibility was estimated to 98-100% and specificity to 93-100%.
(Sorensen V, 1999)

MATERIAL AND METHOD

There were analized 90 serum samples from porcine 4-5 months old in two
breeding units.
From each animal were taken 10 ml blood punctioning the jugular
confluent, in vacutainers without anticoagulant.
The blood samples were maintained to room temperature for 1 hour and
after that the coagul was detached from the tubes walls with a metalic shaft and
the samples were introduced in the refrigerator (+40C, 24h) for the complete
serum expression. The serum samples were transfered in sterile Eppendorf
tubes.
For ELISA it was used DakoCytomation kit (wich contains Microtitration
Plate, White Streeps, Sealing Tapes, Sample Diluent, Positive Control Serum,
Negative Control Serum, Conjugate, Wash Buffer Concentrate, Substrate, Stop
Solution), ELISA NUNC Washer and ELISA Rosys Athos 2010 Reader.
Porcine serum samples were diluted 1:10 and incubated for 90 minutes in
microwells pre - coated with M. hyopneumoniae antigen. Peroxidase conjugated
monoclonal antibody to M. hyopneumoniae is then added to the wells containig
the diduted samples. The conjugated antibody competes with any M.
hyopneumoniae antibody present in the porcine serum for binding epitopes on the
immobilised antigen. n all the wells a yellow-brown colour appears. After 15
minutes, wells are washed and a chromogenic substrate is added. The color
reaction is stopped by the acid addition.
The higher the antibody concentration in the porcine serum is, the less
conjugate monoclonal antibody is able to bind to the immobilised antigen,
resulting in a lower colour intensity. Conversly in porcine samples containing low
levels or no antibody, more conjugate monoclonal antibody is able to bind,
resulting in a a higher colour intensity.
Results are read at 450nm in Optical Density Units (OD) and samples OD
is compared with the measure one (Buffer solution).
Positive samples present OD values 50% lower then buffer solution OD.

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RESULTS AND DISCUTIONS

Serological screening of swine from the analysed breeding units was


realized for M. hyopneumoniae wearers identification.
Results interpretations was made having as a measure Buffer control
optical density value (BCOD). The results are considered corect if there are
performed the following critera: BSOD values between 0,750 and 2,500, Positive

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Control Serum OD 50% of the BCOD (PCOD) and Negative Control Serum
OD 75% of the BCOD (NCOD).
The obtained results are:
- BCOD 2,043 (Buffer control)
- PCOD 0,730 (Positive control)
- NCOD 2,032 (Negative control)
In function of BCOD, were considered:
- positive samples 50% of the BCOD (1,021)
- negative samples 65% of the BCOD (1,327)
- doubptful samples = 1,021 - 1,327 (require repetition after 2 weeks).
The obtained values after the reading were systematized in tables 1 and 2.
Table 1
Serological examination results in each of the two breeding units
rapported to total animal number

First unit animals number % Second unit animal number %


Positives 17 37,7 25 55,5
Negatives 20 44,4 8 19,9
Doubtful 8 17,9 12 26,6
TOTAL 45 100 45 100

Table 2
Rezultats obtained after the serological exam on the total animal number

Animal number %
Pozitives 42 46,6
Negatives 28 31,2
Doubtful 20 22,2
TOTAL 90 100

The animals from the first unit, with imported porcine didnt presented
clinical signs of disease, the aim of this screening being the establishing of M.
hyopneumoniae wearers between those animals.
The animals from the second breeding unit, with native porcine, presented
clinical signs (dry cough, dispneic respiration).
The obtained results showed that M. hyopneumonie specific antibodies are
present in a number of 42 serum samples (46,6%) from the total of the 90
examined, 17 samples (37,7%) deriving from a breeding unit with imported
porcine and the other 25 samples (55,5%) from a plot of native porcine. Because
of the lack of clinical signs in the animal from the first unit, must be considered
the wearer state of those animals after healing (a 66 weeks periode) (Perianu
Tudor, 1996) or an ocult infection.
The animals with doubtful results must be retested after 2 weeks, using the
same method, but after various studies through this assay, it was established that

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this doubtful results can be included in the negative category in the statistical
interpretation.
Being a sensitive test, which can discriminate M. hyopneumoniae specific
antigenes, the screening values must be considered.
The serology in M. hyopneumoniae enzootic pneumonia present different
limites, the seroconversion after the infection being, generaly, delayed and
various. (Sitjar and col., 1996; Calsamiglia and col., 1999a), those data suggesting
that the serological results extrapolation to clinical infection is difficult. In
infected effectives, mortality rate can reach 1 - 5%, lower then morbidity, which
can reach 40 - 60%.
M. hyopneumoniae actions in high density effectives, inproper ventilation,
high level of ammonia in atmosphere, defficient sanitary conditions, maintaining
of various ages animales in the same place, increasing the possibility of horizontal
transmition. (Marco Ibarra C.. and col. 2000).
Literature data confirmes our results, 90% of the Belgic porcine effectives
being infected. In SUA, morbidity in adult animals is of 90-100% and of the
youth is of 74-85% (Bush Eric, 2002). Respiratory problems are frequent in all the
countries with intensive porcine breeding. There are affected over then 97% of
animals, more of the 70% presenting specific lesions (Davies and col., 1993).

CONCLUSIONS

For M. hyopneumoniae specific antibodies detection were analized 90


serum samples from two porcine breeding units.
From the first unit with imported porcine, the number of positive samples
was of 17 (37,7%).
In the second unit with native porcine, the number of positive samples was
of 25 (55,5%), corelated with clinical signs presence.
From the total of 90 analized sample serums 46,6% were positive,
concluding that M. hyopneumoniae wearing is very high in porcine from import
as well as in the native ones breeded in industrial system.
The presence of M. hyopneumoniae specific antibodies in unvaccinated
animals, signifies un ocult infection or the wearer state.

BIBLIOGRAPHY

1. Bush E., 2002 - The epidemiology of Mycoplasma hyopneumoniae, Swine


Mycoplasma Pneumonia Workshop, Kansas City, MO, March 6
2. Carp-Crare M., Dorina Timofte, 2002 Immunology and Immunopatology, Venus
Pub,lishing House, Iai;
3. Davies P.R., Dial G.D., Marsh W.E., Bahnson P.B., 1993 - Feasibility of
implementing a national swine slaughter monitoring system for the collection of
health data from American swine heards. University of Minnesota;

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

4. Feld N.C., Qvist P., Ahrens P., Friis N.F., Meyling A., 1992 - A monoclonal
blocking ELISA detecting serum antibodies to Mycoplasma hyopneumoniae,
Veterinary Microbiology. , 30(1):35-46;
5. Ibarra M., Norma No M., Arnaldo Alvarado S. Rosa Perales C., 2000 -
Evidencia de la presencia de anticuerpos de Mycoplasma hyopneumoniae en
cerdos provenientes de granjas de crianza artesanal del sur de Lima, Rev Inv
Vet Per; 11(2):164-168;
6. Kobisch M., N.F. Friis, 1996 - Swine mycoplasmoses, Rev Sci Tech, 15(4):1569-
605;
7. Perianu T., 1996 Animals Infectious diseases, Bacteriozis, vol. I, Ed. Venus Iai;
8. Sorensen V., Barfod K., Feld N.C., 1999 - Evaluation of a monoclonal blocking
ELISA and IHA for antibodies to Mycoplasma hyopneumoniae in SPF-pig herds ,
The Veterinary Record, Vol. 130, Issue 22, 488-490;
9. Sitjar M., Noyes E., Moreso J.M., Fernadnez de Aragon J., Pijoan C., 1996 -
Relationship among seroconversion to Mycoplasma hyopneumoniae, lung
lesions, and production parameters in pigs, Swine Hlth Prod. 4:273-277.

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EVALUARE EPIDEMIOLOGIC I ECONOMIC A


PROGRAMULUI DE COMBATERE A TUBERCULOZEI
BOVINE N ROMNIA
EPIDEMIOLOGICAL AND ECONOMICAL EVALUATION OF THE ERADICATION
PROGRAM IN BOVINE TUBERCULOSIS IN ROMANIA

Gh. SAVUA1, A. STOICHICI2,


D. ANI1, Adriana ANI1,
C. BUTUNOI1, Roxana POPOVICI-BIBIRU1
1
U.S.A.M.V. Iai
2
I.D.S.A. Bucureti

In this study we are presenting an epidemiological and economical


analisis of the strategic surveillance program of bovine tuberculosis in
Romania.
Based on the analises of the national oficial data raported to OIE
and of the sanitary evidences from three counties in Romania, we ascertain
a constant tendency to the reduction of the incidence of the tuberculosis
infection in Romania after 1900, this beeing situated at the amount of
0,04% in 2004. In these circumstances the exhaustive testing of the bovine
older than six weeks, action that costs aproximativelly 10 mil. EUR
annually is no longer justificated.

Key words: bovine tuberculosis, diagnosis, epidemiology, economy,

Tuberculoza bovin este o bacterioz cronic ce afecteaz animale


domestice i slbatice, ndeosebi bovidee i este transmisibil la om prin
intermediul laptelui. Este produs de Mycobacterium bovis i constituie nc o
grav problem de sntate n multe ri din lume. (1, 3, 4).
Infecia tuberculoas la bovinele n via se depisteaz, n mod obinuit,
prin testarea hipersensibilitii specifice de tip ntrziat (test alergic intradermic)
iar mai recent s-au pus la punct tehnici bazate pe evidenierea diferiilor markeri
din snge (testul de proliferare a limfocitelor, testul gamma-interferonului, testul
imunoenzimatic etc), teste a cror valori intrinseci de diagnostic, sensibilitatea i
specificitatea, mai trebuie ameliorate i ale cror costuri sunt nc ridicate pentru a
deveni accesibile pe scar larg. (9).
Dup moarte ori sacrificare, diagnosticul se confirm prin examene
anatomopatologice, histopatologice, bacterioscopice i bacteriologice. Izolarea i
identificarea bacilului tuberculos specific, prin metode biochimice i genetice
(PCR) constuie metodele de certitudine. (2).
Lupta mpotriva acestei boli se axeaz pe depistarea ct mai timpurie a
animalelor infectate i eliminarea sistematic a acestora din efective, pe baza unor

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programe strategice adecvate i adaptate prevalenei infeciei i circumstanelor


epidemiologice specifice ale fiecrei ri sau chiar regiuni. (8, 9).
Ca i n multe alte boli, aceste programe de combatere se doresc a fi
eficiente att din punct de vedere epidemiologic ct i economic. (6).
Pentru a interveni eficient i cu justee n combaterea i limitarea
rspndirii unei boli, trebuie ca fiecare aciune de sntate animal, de depistare a
unui fenomen epidemiologic s cuprind i un bilan economic, viznd att
costurile directe ct i cele indirecte, precum i costurile sociale pe care le implic
acel program. (5).

MATERIAL I METODE

n vederea realizrii acestui studiu s-au utilizat informaii din bazele de date
raportate i publicate la Oficiului International de Epizootii (OIE) precum i din
evidenele primare ale serviciilor veterinare din trei judee ale Romniei, n limita
posibilitii accesului la acestea i n condiiile unei neconcordane formale ntre
diversele organisme i trepte erarhice n ceea ce privete modalitatea de
nregistrare, prelucrare primar i de raportare a datelor.
Rezultatele investigaiilor nu au fost corelate ntre cele trei judee,
urmrindu-se doar evidenierea nivelului prevalenei tuberculozei bovine i
evaluarea economic brut a eforturilor cuantificabile fcute n cadrul aciunilor de
supraveghere i de combatere a acestei boli.
Datele au fost sintetizate i prezentate sub form de tabele iar evaluarea
economic s-a facut lund drept reper preurile medii anuale ale diferitelor
manopere i consumabile, fr o actualizare strict din punct de vedere financiar.
S-a urmrit, n primul rnd, evidenierea grandorii efortului fcut n virtutea
programului strategic naional de supraveghere i control a bolilor la animale i
aprecierea justeei unei astfel de strategii, n condiiile date de frecven a
tuberculozei bovine n Romnia.

REZULTATE I DISCUII

Din informaiile raportate i nregistrate n baza de date a Oficiului


International de Epizootii, referitor la incidena tuberculozei bovine n Romnia n
perioada 1996-2004, s-au putut sintetiza i analiza aspecte privind evoluia
focarelor, a cazurilor i a msurilor de combatere aplicate (Tabelul 1).
Fa de o populaie de referin aproape constant anual, care oscileaz n
jurul unui efectiv de aproximativ trei milioane bovine, n perioada 1996-2004 se
observ o tendin staionar a focalitii tuberculozei bovine (ntre 30 i 163 de
focare anual) i o scdere a reactivitii individuale a bovinelor la testul alergic, de
la 22 774 de reacii pozitive n 1997 la 74 cazuri n 2002, cu o uoar cretere n
2003 i 2004.

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Tabelul 1
Incidena anual a tuberculozei bovine n Romnia n perioada 1996-2004

Populaia de Cazuri Inciden Animale


Focare
Anul referin reagente cazuri sacrificate
(nr.)
(mii cap.) (nr.) (%) (nr.)
1996 3 496 79 11 700 0,335 6 600
1997 3 436 163 22 774 0,663 8 112
1998 3 236 71 304 0,009 141
1999 3 143 46 82 0,003 61
2000 3 061 68 130 0,004 61
2001 2 870 30 361 0,013 297
2002 2 800 30 74 0,003 1 901
2003 2 878 113 786 0,027 1 481
2004 2 897 148 1 099 0,038 541

Apreciind dinamica rspunsurilor pozitive n valori procentuale, deci


frecvena relativ a fenomenului, se constat valori cuprinse ntre 0,33% n 1996
i 0,003% n 2002, ceea ce situeaz Romnia pe o poziie foarte bun din punct de
vedere epidemiologic privind tuberculoza bovin.
Este de remarcat scderea progresiv a cazurilor de tuberculoz bovin
dup anii 1990, ceea ce valideaz reuita strategiei de pn n acest moment n
combaterea tuberculozei bovine. (7).
n continuare se pune problema meninerii statutului de indemnitate a
efectivelor de bovine n condiii de mare eficien epidemiologic dar i
economic, tiut fiind faptul c tuberculinarea reprezint o aciune accesibil dar
care implic i costuri cu materialele i manopera, ca s nu mai vorbim despre
stress-ul provocat animalelor dar i de inconvenientele induse proprietarilor.
Pentru a ilustra mai bine i pentru a putea susine ideea eficientizrii
economice a luptei mpotriva tuberculozei bovine, am efectuat scurte analize ale
frecvenei infeciei n trei judee, B,V i N, pe parcursul unor perioade
diferite de timp dar sugestive ca situaie epidemiologic.
Tabelul 2
Frecvena tuberculozei bovine n judeul B n perioada 2002- 2005

Populatia de Numar total de


Animale Cazuri pozitive
Anul referinta animale
reagente (confirmate)
(nr. bovine) tuberculinate
2002 77 556 58 630 754 13
2003 73 769 70 923 386 16
2004 76 272 60 060 377 22
2005 75 671 62 024 363 44

Din datele prezentate n tabelul nr. 2 se observ c la nivelul judeului B


proporia de animale reagente are o tendin descresctoare n timp, de la 1,29%

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la 0,58% din animalele testate dar numrul cazurilor confirmate este n uoar
cretere (de la 13 cazuri, n 2002, la 44 de cazuri n 2005). Din investigaiile
noastre am reinut faptul c aceste cazuri provin ndeosebi din gospodriile
individuale, ceea ce sugereaz un risc al sistemului extensiv de cretere a
bovinelor, nc dominant n regiune.
Tabelul 3
Frecvena tuberculozei bovine n judeul V n perioada 2003 - 2005

Populatia de Numar animale Nr. operatii Nr. Cazuri


Anul referinta tuberculinabile tuberculinare anim. pozitive
(nr. bovine) (varsta >6 sapt.) (total /an) reagente (confirmate)
2003 74 966 68 258 141 126 810 10
2004 77 274 76 123 148 666 795 7
2005 85 358 80 891 187 539 696 3

n judeul V (tabelul 3) se observ o tendin descresctoare a numrului


de reacii pozitive la testul alergic ns i o scdere a numrului de cazuri
confirmate, ceea ce este n consonan cu situaia naional prezentat anterior.
Tabelul 4
Frecvena tuberculozei bovine n judeul N n perioada 1995 - 2004

Nr. operaii
Populaia de referin Cazuri pozitive
Anul tuberculinare
(nr. bovine) (confirmate)
(total /an)
1995 84 913 152 660 -
1996 86 863 152 500 2
1997 79 363 143 147 2
1998 82 340 125 210 2
1999 88 030 128 261 -
2000 93 373 138 133 -
2001 92 867 141 395 -
2002 98 461 150 800 -
2003 96 117 171 568 -
2004 97 758 167 517 -

Tabelul 4 prezint o situaie sintetic a efortului de supraveghere a


tuberculozei ntr-un jude n care prevalena infeciei s-a redus practic pn la
zero, timp de ase ani consecutivi nemaifiind nregistrat nici un rspuns pozitiv.
n aceste condiii se pune ntrebarea dac se mai justific testarea semestrial a
tuturor bovinelor n vrst de peste ase sptmni ?
O analiz simpl, dar sugestiv, a costurilor unei aciuni de tuberculinare ne
arat faptul c efortul financiar este apreciabil i poate fi considerat o risip n
condiii de indemnitate fa de tuberculoza bovin a unor efective i chiar zone
mai mari, judee i regiuni.

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Astfel, pe baza informaiilor obinute, costurile aciunii de tuberculinare la


nivelul judeului V, pe parcursul a trei ani consecutivi, se prezint asfel:
Tabelul 5
Evaluarea costurilor aciunii de tubeculinare a bovinelor n judeul V

Nr. total de TOTAL


ANUL Cost /tuberculinare
tuberculinri ( RON )
2003 141 126 5,1060 720 589,356
2004 148 666 6,1145 909 018,257
2005 187 539 7,0330 1 318 961,787

Cifrele din tabelul de mai sus nu mai necesit comentarii. Dac am


extrapola la ntreaga ar valorile medii estimate pentru anul 2005 n acest jude,
am putea evalua efortul financiar ca fiind de aproximativ 35 000 000 RON, adic
echivalentul a 10 000 000 EUR anual.

CONCLUZII

Conform datelor oficiale, incidena tuberculozei bovine n Romnia are o


tendin de scdere continu, ceea ce confirm succesul programului naional de
combatere a acestei boli, program adoptat i aplicat dup 1990.
n acelai timp, se remarc un efort financiar i logistic considerabil n
fiecare an, n condiiile testrii semestriale exhaustive a tuturor bovinelor n vrst
de peste ase sptmni.
n condiiile unei prevalene a tuberculozei bovine care tinde spre zero, un
asemenea efort nu se justific.
Indemnitatea efectivelor se poate menine prin aplicarea unor msuri
sanitare susinute i prin testri periodice prin sondaj, la intervale de timp din ce
n ce mai mari i difereniat pe zone epidemiologice cu statut diferit.

BIBLIOGRAFIE

1. Mnzat Moga R., 2001 - Boli infecioase ale animalelor. Ed. Brumar, Timioara.
2. Paul I., 2005 - Etiomorfopatologia bacteriozelor la animale. Ed. Pim, Iai
3. Perianu T., 2004 - Boli infecioase ale animalelo r- bacterioze. Vol.1, Ed. Venus, Iai.
4. Rpuntean Gh., Rpuntean S., 2005 - Bacteriologie veterinar special, Ed.
Academic Pres, Cluj - Napoca.
5. Savua Gh., 2001 - Epidemiologie veterinar general. Ed. Ion Ionescu de la Brad
Iai.
6. Toma B., Dufour Barbara, Sanaa M., Benet J.J.,Ellis P., Moutou F., Louza A.,
1996 Epidmiologie appliqu a la lutte collective contre les maladies animales
transmissibles majeures. Ed. AEEMA, Maisons-Alfort;
7. *** - Programul aciunilor de supraveghere, prevenire i control al bolilor la animal,
ANSVSA, Bucureti, 2000-2004.
8. *** - O.I.E., Manual de standarde pentru testele de diagnostic i vaccinri,
www.oie.int

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EPIDEMIOLOGIA PARATUBERCULOZEI
RUMEGTOARELOR N ROMNIA
THE EPIDEMIOLOGY OF RUMINANTS PARATUBERCULOSIS IN ROMANIA

Gh. SAVUA, Irina MATEI,


Selma MURAT, D. ANI
U.S.A.M.V. Iai

We have made a retrospective study of the ruminants


paratuberculosis in Romania between 1990 and 2004, based on the
national documents and epidemiological raports and in particulary
raportated to two counties in the East of Romania . From the data raported
to OIE, results that the evolution of the disease had a descendent incidence
in the period of the sudy. But in the two counties studied, the depistation test
made, shows an aparent prevalence wich varies in large limits between
different groups of animals and in time, raported to the test. These
observations show an evident lack of correlation among the data. For the
estimation of the disease real prevalence is necessary to made an
epidemiological investigation based on a scientific method of sampling and
on the concomitent use of more specific and sensible depistation methods.

Key words: paratuberculosis, ruminants, epidemiology,


epidemiological investigation.

Paratuberculoza (boala lui Johne, enterita cronic hipertrofiant) este


produs de ctre Mycobacterium avium, subspecia paratuberculosis (8) i se
caracterizeaz printr-o enterit cronic, cu caracter granulomatos, ce afecteaz
mai multe specii de rumegtoare domestice i slbatice precum i alte specii de
animale de pe tot globul.
Diagnosticul acestei boli se bazeaz pe semnele clinice, coroborate cu
leziunile specifice de tip granulomatos de la nivelul tubului digestiv i se
confirm prin identificarea bacilului lui Johne n fecale i esuturi, prin examen
bacteriologic, histopatologic sau prin PCR (polymerase chain reaction). (6, 10).
Depistarea animalelor infectate subclinic, individual sau la nivel de turm,
constituie ns o problem delicat i ea se bazeaz pe detectarea anticorpilor
specifici prin serologie, pe identificarea bacilului n culturi din fecale i esuturi
sau prin evidenierea imunitii mediate celular.(6).
Testele serologice utilizate curent pentru depistarea animalelor infectate
subclinic sunt: reacia de fixare a complementului (RFC), testul imuno-enzimatic
(ELISA) i imunodifuzia n gel de agar (ID), pentru evidenierea imunitii
umorale i testul gamma-interferonului pentru evidenierea imunitii mediate
celular. Aceasta din urm, respectiv hipersensibilitatea de tip ntrziat, se mai

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evideniaz i prin testul alergic intradermic, n varianta sa unic-TU sau


comparativ simultan - TCS. (6, 9).
Alegerea unora sau combinarea mai multor teste de depistare a
paratuberculozei reprezint decizia esenial pentru controlul infeciei subclinice
i ea trebuie luat n funcie de circumstane i de sensibilitatea i specificitatea
fiecrei metode aplicate la nivel de individ i la nivel de turm. (7, 10).

MATERIAL I METODE

Pentru realizarea prezentei analize, privind incidena i prevalena


paratuberculozei rumegtoarelor din Romnia, s-au utilizat informaii din bazele
de date raportate i publicate la Oficiul Internaional de Epizootii precum i
rezultatele unor studii efectuate separat n dou judee din estul Romniei, B i
C, pe diferite segmente de timp din perioada 1990-2004, funcie de datele
disponibile.
S-a urmrit frecvena infeciei paratuberculoase la bovine i ovine, n
consonan cu strategia de supraveghere aplicat n teritoriu i cu evoluia
reglementrilor n vigoare, att n sectorul extensiv ct i n ferme organizate, pe
baza datelor i nregistrrilor existente n documentele de eviden din cadrul
serviciilor de specialitate i a laboratoarelor sanitare veterinare din cele dou
judee luate n studiu.
Informaiile prelevate din evidenele studiate au fost analizate i interpretate
prin metodele epidemiologice descriptive, fr o comparabilitate ntre ele ci doar
cu intenia de a aprecia gradul de pertinen al strategiei aplicate vis a vis de
situaia raportat .

REZULTATE I DISCUII

Analiznd informaiile nregistrate n baza de date a OIE, privind incidena


paratuberculozei n Romnia pentru perioada 1996-2004, s-au putut identifica
sincope n raportarea situaiei, aa cum rezult din Tabelul 1.
Astfel, dac n 1996 sunt raportate doar trei cazuri de paratuberculoz la
ovine, n urmtorii trei ani, 1997,1998 i 1999, lipsesc informaiile naionale, ceea
ce nu nseamn c boala a disprut din Romnia ci mai degrab faptul c sistemul
de supraveghere din partea serviciilor veterinare trecea printr-o perioad dificil.
Considernd c la nivel naional au fost luate n considerare i au fost
raportate drept cazuri doar cele confirmate etiologic, n urma sacrificrii
animalelor suspecte i a examenelor de laborator, am efectuat o analiz detaliat
a rezultatelor obinute la nivel local n cadrul aciunii de depistare a infeciilor
subclinice cu Mycobacterium paratuberculosis prin testele uzuale.
Ca urmare, la nivelul judeului B, au fost identificate i sistematizate
rezultatele testrilor efectuate la bovine n perioada 1990-2002 prin TCS, RFC si
bacterioscopie (Tabelul 2).

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

Tabelul 1
Incidena anual a paratuberculozei rumegtoarelor n Romnia
n perioada 1996-2004

Nr. Anul Bovine Ovine/Caprine


crt. Focare Cazuri Focare Cazuri
1. 1996 0 0 1 3
2. 1997 ? ? ? ?
3. 1998 ? ? ? ?
4. 1999 ? ? ? ?
5. 2000 2 6 0 0
6. 2001 2 3 1 33
7. 2002 43 149 54 579
8. 2003 52 180 89 6191
9. 2004 2 198 32 762

Tabelul 2
Prevalena aparent a paratuberculozei la bovinele testate prin TCS, RFC i
bacterioscopie pe fecale n judeul B n perioada 1990 2002

T.C.S. R.F.C. BACT.


Anul
Nr. + % Nr. + % Nr. + %
1990 8460 35 0.4 7300 18 0,2 370 2 0,05
1991 7730 20 0,3 6750 13 0,2 250 - -
1992 6660 10 0,2 5700 5 0,1 140 - -
1993 7530 8 0,1 6200 - - 190 - -
1994 7030 3 0,04 6900 2 0,02 150 - -
1995 6815 - - 5700 - - 180 - -
1996 6470 - - 6100 - - 130 - -
1997 6720 - - 6400 - - 72 - -
1998 6530 - - 5840 - - 45 - -
1999 5990 - - 5500 - - - - -
2000 5870 - - 5300 - - - - -
2001 5975 - - 5450 - - - - -
2002 5890 - 4500 - - - - -

Se constat faptul c frecvena rezultatelor pozitive este n scdere vizibil


n perioada 1990-1994, pentru ca din 1995 s nu se mai nregistreze reacii
pozitive la nici unul din testele folosite. Comparnd cele trei metode ntre ele, se
observ c reactivitatea cea mai mare se manifest la testul alergic (TCS), fapt
studiat i prezentat anterior de ctre noi (6) i de ctre ali autori (3, 4, 8).
Efectund aceeai analiz i n cazul ovinelor din judeul B, testate prin
metode identice, am putut sintetiza situaia prezentat n tabelul 3.

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Tabelul 3
Prevalena aparent a paratuberculozei la ovinele testate prin TCS, RFC i
bacterioscopie pe fecale n judeul B n perioada 1990 2002

Anul T.C.S. R.F.C. BACT.


Nr. + % Nr. + % Nr. + %
1990 17452 1470 8,4 12500 289 2,3 95 5 5,3
1991 16250 480 3,0 12300 213 1,7 77 - -
1992 12740 380 2,4 11250 135 1,2 58 3 5,2
1993 15420 430 2.8 10450 147 1,4 60 2 3,3
1994 12494 120 1.0 10650 132 1,2 42 2 4,7
1995 10710 70 0,7 11190 85 0,8 27 - -
1996 9380 82 0,9 12000 70 0,6 13 - -
1997 9500 37 0,4 11750 105 0,9 27 - -
1998 9250 42 0,5 10500 98 0,9 18 - -
1999 12030 25 0,2 10300 180 0,7 21 - -
2000 10100 18 0,2 9100 58 0,6 35 - -
2001 9140 - - 8120 7 0,08 28 - -
2002 8500 - - 7500 - - 17 - -

n cazul ovinelor se constat o frecven mai mare a rezultatelor pozitive la


toate cele trei metode utilizate, cu un procent de pn la 8,4% n cazul testului
alergic i cu existena de reacii pozitive, n descretere ns, pn n anul 2000,
dup care efectivele se negativeaz. Acest aspect este explicabil dac lum n
considerare faptul c n fiecare an exemplarele reagente au fost eliminate,
conform programului naional strategic de supraveghere i combatere a bolilor
transmisibile ale animalelor. (9).
Analiznd situaia epidemiologic a paratuberculozei la bovine n judeul
C, pe baza rezultatelor obinute la aceleai teste aplicate n perioada studiat s-a
constatat o frecven a reaciilor pozitive la testul alergic ce a variat ntre 0,81% i
5,26% la diferite efective controlate n anul 2000 i ntre 0,76% i 2,88% n anul
2002. Examenele serologice (RFC) aplicate n aceleai efective au relevat o
frecven a rspunsurilor pozitive cuprins ntre 0% i 22,64% (??) n anul 2000 i
ntre 0,5% i 11,9% n anul 2002, ceea ce ilustreaz nc odat necorelarea
reactivitii umorale cu cea celular i relativitatea testelor utilizate, ndeosebi a
RFC, test cu o sensibilitate redus i de multe ori variabil.
Referitor la frecvena infeciei paratuberculoase la ovinele din acelai jude,
am remarcat doar situaia dintr-o ferm contaminat i supus unui program de
asanare, procentul reaciilor pozitive la TCS fiind de 12% n anul 2000 i de
10,5% n anul 2002 iar al rezultatelor pozitive la RFC fiind de 16% n anul 2000
i de 10,5% n anul 2002 .
Toate aceste rezultate nregistrate la nivel judeean nu se coreleaz, n final,
cu situaia raportat la nivel naional, ceea ce ne face s apreciem cu nencredere
rezultatele aciunii de supraveghere a acestei boli n ara noastr i s afirmm

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faptul c prevalena real a paratuberculozei este nendoielnic alta dect cea


consemnat n rapoarte.

CONCLUZII

Analiza acestor rezultate scoate n eviden prezena i evoluia


paratuberculozei rumegtoarelor n Romnia, cu o tendin de scdere a
frecvenei, conform datelor oficiale.
Metodele de depistare i de diagnostic folosite nu sunt suficient de sensibile
i specifice, lipsind ndeosebi metodele de identificare cert a agentului patogen
(examenul bacteriologic, PCR etc.).
Rezultatele obinute prin diferitele teste aplicate acelorai efective nu se
coreleaz ntre ele, ceea ce face ca prevalena real s fie greu de apreciat.
Relativitatea rezultatelor obinute prin testele aplicate i deficienele n
aplicarea programului de supraveghere i n raportarea datelor au generat
neconcordane ntre diferitele situaii epidemiologice analizate.
Se impune un studiu epidemiologic de prevalen a paratuberculozei
rumegtoarelor n Romnia bazat pe utilizarea unei metodologii tiinifice de
eantionare i prin utilizarea concomitent a metodelor directe (etiologice) i
indirecte (serologice i alergice) de depistare, a cror sensibilitate i specificitate
s fie luate n calcul la interpretarea rezultatelor.

BIBLIOGRAFIE

1. Carp-Crare M., 1998 - Metodologii de diagnostic imunologic. Ed. Ion Ionescu de la


brad, Iai.
2. Mnzat Moga R., 2001 - Boli infecioase ale animalelor. Ed. Brumar, Timioara.
3. Paul I., 2005 - Etiomorfopatologia bacteriozelor la animale. Ed. Pim, Iai.
4. Perianu T., 2004 - Boli infecioase ale animalelo r- bacterioze. Vol.1, Ed. Venus, Iai.
5. Rpuntean Gh., Rpuntean S., 2005- Bacteriologie veterinar special, Ed.
Academic Pres, Cluj - Napoca.
6. Savua Gh., 2000 Paratuberculoza rumegtoarelor. Ed. Ion Ionescu de la Brad Iai
7. Savua Gh., 2001 - Epidemiologie veterinar general. Ed. Ion Ionescu de la Brad
Iai.
8. Thorel Marie-Francoise, 1990 Recherches taxonomiques et genotipiques sur les
mycobacteries mycobactines-dependentes. These Doct. dUniv.,Universite Paris
XII,158
9. *** Programul aciunilor de supraveghere, prevenire i control al bolilor la animale,
ANSVSA, Bucureti, 2000-2004.
10. *** O.I.E., Manual de standarde pentru testele de diagnostic i vaccinri, www.oie.int.

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MODIFICAREA STRUCTURII BURSEI LUI FABRICIUS


SUB ACIUNEA TULPINILOR VACCINALE FIERBINI
CONTRA BURSITEI INFECIOASE AVIARE
THE ACTIONS OF HOT STRAIN BG AGAINST IBD TO STRUCTURAL
MODIFICATIONS OF TISSUES OF CLOACAL BURSA

N. STARCIUC, I. SCUTARU,
S. ZELENIN
U.A.S. Republica Moldova

Meninerea efectivelor de psri n stare epizootic satisfctoare att n


condiiile industriale ct i a sectoarelor particulare se datoreaz faptului utilizrii
mijloacelor specifice de profilaxie a bolilor infecioase.
Actual se folosete o gam destul de mare de vaccinuri n prevenirea bolilor
infecioase la psri, dintre care o importan mai deosebit le revine vaccinurilor
contra pseudopestei aviare, bronitei infecioase, bursitei infecioase.
Comparativ cu aciunea altor vaccinuri cele utilizate n scopul prevenirii
bursitei infecioase aviare (BIA) este deosebit. n funcie de gradul de
reactogenitate vaccinurile contra BIA sunt clasate ca tulpini moi, intermediare i
fierbini. Toate aceste tulpini vaccinale provoac modificri structurale ale
esuturilor bursei lui Fabricius care difer n funcie de gradul de reactogenitate.
Scopul cercetrilor a fost de a stabili deosebirile modificrilor esutului
bursei lui Fabricius sub aciunea tulpinei vaccinale fierbinte BGcontra BIA.

MATERIAL I METOD

Cercetrile au fost efectuate pe un lot de 50 capete pui de gin care au


fost vaccinate contra BIA cu tulpina fierbinte, BG de dou ori la vrsta 10 i 20
zile prin adpare i respectiv grupa lot martor.
Pn la vaccinare i a 10, 20, 35 i 45 zile dup vaccinare, din fiecare
grup sau sacrificat cte 5 capete de pui, de la care sa recoltat probele de snge
i bursa lui Fabricius pentru examenul histologic.
Preparatele histologice sau vopsit cu hematoxilin i eozin, structura
esuturilor bursei lui Fabricius s-a examinat la microscopul cu obiectivul 40 i
ocularul 12,5.

REZULTATELE OBINUTE

La puii din grupul lot martor epiteliul bursei lui Fabricius este
pluristratificat, cilindric. n esutul epitelial al submucoasei se observ plasmocite
unice. Foliculii limfoizi nu prezint hiperplazie. n unii foliculi predomin

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

prezena limfocitelor i a limfoblastelor, n centrul se observ un numr mic de


macrofage.
n esutul burselor puilor din grupa experimental unde au fost vaccinai cu
tulpina BG, la a 10-a i 20-a zi dup a doua vaccinare, bursa lui Fabricius era
constituit din foliculi mici i medii. n esutul perifolicular se obsevau fenomene
infiltrative limfohistocitare, exudaii i unele celule necrotizate, care de regul
sunt localizate la periferiile foliculilor.
La a 35-a i 45-a zi dup vaccinare foliculii limfoizi sunt de diferite
dimensiuni cu predominarea celor medii i mici. n majoritate foliculilor se
observ fenomene infiltrative, focare necrotice, procese fibroplazice cu tendin
de nlocuire a esutului limfoid propriu-zis sau formarea unor chisturi.

CONCLUZII

Vaccinul fierbinte din tulpina BG utilizat n scopul prevenirii BIA,


provoac unele modificri inflamatorii distinctive n foliculii bursei lui Fabricius
ce prin urmare duce la o stare de imunosupresie la pui.

BIBLIOGRAFIE

1. Hassan M.K., Saif Y.M., 1996 - Influence of the host sistem on the pathogenicity,
immunogenicity and antigeniticy of infections bursal disease viruses. Avian Dis.,
40 : 553-561
2. Sivanandon V.Y., Sasipreeyajan D.A., Halvarson, Newman Y.A., 1986 -
Histopathologic changes indused by serotype infectious bursal disease virus in
specific- pathogen- free chickens. Avion Dis., 30 : 709-715
3. Colnek B.W. - Diseases of poultry. Tenth edition. Infectious Bursal Disease. Iawa
State University Press. USA. p. 721-760

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ASPECTE EPIDEMIOLOGICE PRIVIND BRUCELOZA


OVIN (INFECTIA CU BRUCELLA OVIS)
N JUDEUL VASLUI

THE EPIDEMIOLOGICAL ASPECTS ABOUT THE INFECTION


WITH BRUCELLA OVIS IN VASLUI COUNTY

C. NECULI1, M. CARP CRARE2,


Narcisa NECULI1, Adriana CHIRIAC3
1
D.S.V.S.A. Vaslui
2
U.S.A.M.V. Iai
3
L.S.V.J. Vaslui

During 2000-2005, an epidemiological study was effected on the


rams in Vaslui county, through processing of dates concerning the evolution
of infection with Brucella ovis.
The researches were effected on the blood samples coming from the
rams on Vaslui county, processed through the complement fixation test
(CFT). We established the incidence and the prevalence of the infection
with Brucella ovis in six years of study.
This way, in Vaslui county, the biggest incidence was in 2000 (123
cases - 1,33%) and in 2002 (155 cases - 1,46%). During 2000-2005, the
incidence of infection with Brucella ovis was 8,97 %.
In this teritory, the prevalence of this disease was 0,20%.The
evolution of Brucella ovis in Vaslui county between 2000 and 2005, had a
relatively decrease of its incidence.

Key words: Brucella ovis, diagnosis, the complement fixation test, incidence,
prevalence.

Brucella ovis produce infecii clinice sau subclinice la ovine caracterizate


prin leziuni genitale la berbeci i infecii placentare la oi (6). Consecina
principal a bolii o reprezint reducerea fertilitii la berbeci, avorturi periodice la
oi i creterea mortalitii perinatale (1). Conform programului strategic anual,
este prevzut efectuarea a 2 testri serologice pe an, la berbecii de reproducie,
cu dou sptmni nainte de mont i dup mont, n perioada octombrie
noiembrie.

MATERIALE I METODE

Examenele de supraveghere s-au realizat prin reacia de fixare a


complementului (RFC), (2, 3, 4). S-au prelucrat i valorificat o serie de date
multianuale referitoare la evoluia brucelozei la berbeci n judeul Vaslui n
perioada 2000-2005. Cercetrile s-au efectuat pe probele de snge provenite de

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la berbecii de reproducie din jude. Rezultatele obinute au fost sintetizate n


tabelul 3.

REZULTATE SI DISCUTII

Analiznd activitatea de supraveghere a brucelozei la berbeci finalizat prin


examene de laborator i de combatere prin eliminarea animalelor pozitive, se
constat c un procent mic (de 19,85%) din animalele depistate pozitiv la testul
serologic prin RFC au fost tiate pentru precizare de diagnostic.
Aprecierea activitii de supraveghere sub raportul brucelozei la berbeci
prin testul serologic se realizeaz prin stabilirea ratei examenelor serologice. Rata
examenelor serologice a fost calculat prin raportul dintre efectivul de berbeci
existent i efectivul de berbeci controlat, comparat cu cifra de referin 2 (2 testri
serologice pe an): tabelul 3.
Prin prelucrarea datelor s-au putut stabili incidena si prevalena real (5) a
bolii n cei 6 ani luai n studiu (tabelul 1 i 2).
Tabelul 1
Incidena brucelozei la berbeci n judeul Vaslui n perioada 2000-2005

Efective Animale mbolnvite Incidenta


Anul
berbeci n anul curent anuala
2000 4 836 123 123
2001 5 567 33 33
2002 5 448 155 155
2003 5 616 71 71
2004 5 765 50 50
2005 6 158 2 2
Centralizare 33 390 434

Tabelul 2
Prevalena brucelozei la berbeci n judeul Vaslui n perioada 2000-2005

Efective Probe Prevalena


Anul
berbeci confirmate anual
2000 4 836 17 134
2001 5 567 2 50
2002 5 448 23 157
2003 5 616 10 94
2004 5 765 4 60
2005 6 158 1 6
Total 33 390 57

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Pr = a+c/ n% = 57 + 10 / 33 390 % = 0, 20 %
n = nr. total de animale = 33 390 b = fals pozitive; b = 434 - 57 = 377
a = real pozitive; a = 57 c = fals negative; c = 10
d = real negative; d = 33 390 - 434 = 32 95

Datele utilizate n calculul incidenei i prevalenei provin din:


- rezultatele activitii de supraveghere a brucelozei la berbeci n judeul
Vaslui n perioada 2000-2005 (tabelul 3);
- rezultatele activitii de supraveghere prin metode de laborator a
brucelozei la berbeci (tabelul 4.);
Conform tabelului 3, n perioada 2000-2005 s-au sacrificat n judeul Vaslui
57 de berbeci serologic pozitivi.
Tabelul 3
Rezultatele activitii de supraveghere a brucelozei la berbeci
in judeul Vaslui n perioada 2000-2005

Nr. Specificare U/M 2000 2001 2002 2003 2004 2005 Total
crt. Efectiv ovine cap. 222475 225086 212226 216226 225533 238238 1340209
Efectiv
1 existent cap. 4836 5567 5448 5616 5765 6158 33390
berbeci
Efectiv
2 cap. 9235 10697 10612 10755 10816 12111 64226
Examen supraveghere

controlat
Rata
3 2 1,91 1,92 1,95 1,92 1,88 1,97 1,92
controalelor
Numr cap. 123 33 155 71 50 2 434
berbeci
4 depistati
pozitiv la % 1,33 0,31 1,46 0,66 0,46 0,02 0,68
RFC
5 Numr cap. 22 6 30 13 9 2 82
berbeci
sacrificati
6 pentru % 17,89 18,18 19,36 18,31 18,00 100 18,89
diagnostic
Total pozitive
7 confirmate cap. 17 2 23 10 4 1 57
dup sacrificare

Dintre probele examinate serologic prin RFC (7) i cu rezultat de


anticomplementaritate (anume 45), s-au confirmat prin metode de laborator
(examen bacteriologic) un numr de 10 capete (tabelul 4), ce constituie elementul
de calcul pentru reaciile fals negative (c).
I = nabt6 / nast0 %
nabt6= nr. animale mbolnvite n cei 6 ani;
nast0= nr. animale sntoase la nceputul perioadei de observaie.
I = 434 / 4 836 = 8, 97 %

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Tabelul 4
Rezultatele activitii de supraveghere de laborator a brucelozei la berbeci
in judeul Vaslui n perioada 2000-2005

Retestare
Rezultat examen
examinate
Nr. probe

serologica Rezultat final


serologic
Anul La 21 de zile

P D Ac N P D Ac N P D Ac N

2000 9235 123 2 - 9110 - - - - 123 2 - 9110


2001 10697 30 1 3 10663 3 - - - 33 1 - 10663
2002 10612 148 - 27 10437 7 - - 20 155 - - 10457
2003 10755 71 - - 10684 - - - - 71 - - 10684
2004 10816 50 - 15 10751 - - - 15 50 - - 10766
2005 12111 2 - - 12109 - - - - 2 - - 12109
Total 64226 424 3 45 63754 10 - - 35 434 3 63774

CONCLUZII

n judeul Vaslui, n perioada analizat, att incidena ct i prevalena real


a bolii au fost sczute, cu posibiliti reale de asanare a teritoriului din punct de
vedere al infeciei cu Brucella ovis.
Prevalena real a bolii fost foarte redus n teritoriul analizat (0,20 %).
n perioada 2000-2005, datele prelucrate au permis stabilirea unei incidene
a infeciei cu Brucella ovis n judeul Vaslui de 8,97 %, ceea ce creaz premisele
unei asanri rapide a teritoriului.
Incidena cea mai mare a cazurilor depistate serologic a fost n anii 2000
(de 123 cazuri) i 2002 (de 155 cazuri), media multianual a cazurilor de
bruceloz la berbeci fiind de 72,33. S-a observat o rat medie de supraveghere
prin examene serologice pe parcursul celor 6 ani analizai de 1,92, cu minima de
1,88 n anul 2004 i maxima de 1,97 n anul 2005.

BIBLIOGRAFIE

1. American Sheep Industry, U.S.A., Denver, Colorado, 1996 - Chapter ,,Health


,,Sheep production handbook, pag. 350-351
2. Carp-Crare M., Eleonora Guguianu, Dorina Timofte, 1997 - Lucrri practice de
microbiologie veterinar, Uz intern, U.A.M.V. Iai.
3. One E., Virginia Constantinescu, 1978 - Diagnosticul de laborator n medicina
veterinar, Ed. Ceres, Bucureti, 103-127.
4. Rducnescu H., Bica-Popii Valeria, 1986 Bacteriologie veterinar, Editura
Ceres, Bucureti
5. Svua Gh., 2001 - Epidemiologie veterinar general Editura ,,Ion Ionescu de la
Brad, Iai.

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Universitatea de tiine Agricole i Medicin Veterinar Iai

6. *** Border Disease in a Flock of Sheep: Epidemiology, Laboratory and Clinical


Findings (1992) - Journal of American Veterinary Medical Association, 189 (1),
61- 66.
7. *** Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, 5 th edition,
2004, Part 2, section 2.4., chapter 2.4.1.
8. *** Programul aciunilor de supraveghere, prevenire i control al bolilor la animale, al
celor transmisibile de la animale la om, protecia animalelor i protecia mediului
pentru anul 2006.

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REAL-TIME PCR ASSAY WITH TAQMAN PROBE TO


DETECT THE TLH GENE OF V. PARAHAEMOLYTICUS
Gh. BRDAN
U.S.A.M.V. Iai
B. CHINA, G. DAUBE
Universitatea Liege, Belgia

Importance of the fast detection of V: parahaemolyticus (V.p.) is


related to the bacteriological risk associated the consumption of the row or
undercooked seafood. The traditional bacteriological methods are long and
vague; in many cases being no conclusive. This work forms part of a series
of research directed for the development of a routine method, rapid and
reliable, of detection and quantitative appreciation of V.p. by the PCR in
real time, with a TaqMan probe, by using the gene tlh like target. This
established communication that the lower limit of detection, the sensitivity,
is of a bacterium per ml culture. By carrying out a standard threshold of
quantitative appreciation of V. parahamolyticus it is possible to make a
sorting of the samples to continue the research only for the honest ones
which exceeds this limit.

Key Words: detection, real time PCR, V. parahaemolyticus, tlh gene.

INTRODUCTION

The genus Vibrio includes Gram-negative bacilli, anaerobic facultative,


oxidase positive and mobile (10). These bacteria are halophylic meaning that
NaCl stimulates their growth. Among this genus, three species were responsible
for food poisoning due to the ingestion of seafood, V. cholerae the cholera agent,
V. vulnificus responsible for septicaemia and V. parahaemolyticus (V.p )
responsible for gastroenteritidis (5). The clinical signs of V.p. intoxication are:
nausea, fever, vomiting, and headaches. All V.p. isolates produce a thermolabile
haemolysin (TLH). The tlh gene was used as a target for specific real-time PCR in
order to detect V.p. in seafood (9).
The objective of this work is to check the specificity and the efficiency of
the detection of V.p. by real-time PCR, using like target a sequence of nucleotides
of the tlh gene.

MATERIAL AND METHODS

Bacterial strains and microbiological methods. Like bacterial culture


one used the stock of collection ATCC 17.802 (LMG 2850) of V.p., which was
revivified by a 24 hours passage in 10 ml APW 3% salt. The bacteriological

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handling carried out at the time of this project accurately respected the indications
of the standardized method ISO-8914 (1) which describes the method of routine
for the research of the vibrios in the products intended for human consumption by
enrichment. Checking of the purity of the bacterial stock was carried out by
successive passages to 370C, 24h: peptone water salt, selective mediums : TCBS
(agar with thiosulfate, citrate, bile and sugar) and CHROMagar medium.
Characteristic colonies were mended on agar medium nutritive salted and then
tested with the biochemical oxydase and identified with API gallery 20NE
(BioMrieux, Paris, France).
DNA extraction and purification for PCR. The samples of DNA were
obtained after extraction by respecting the indications of the Wizard Genomic
DNA purification kit (Promega Corporation, Madison, WI, USA).
Selection of probe sequences. The sequence of nucleotides was
obtained with the BlastN software of Genbank (8). The complementary and
specific primers oligonuclotides of specific DNA sequence of the tlh gene of V.p.
were worked by Eurogentec (Liege, Belgium): 5' - CGT CAG CGT TGT GAA GCA
A - 3' and 5' - GTC AGC GGC GAA GAA CGT A - 3'. The sequence of tlh probe
was 5' TAG ATT TGG CGA ACG AGA ACG CAG ACA-3 ';
Conditions of real time PCR. The basic equipment to carry out the real-
time PCR was ABI PRISM 7000 thermocycler (Applied Biosystem), with his
software ABI PRISM 7000 SDS (2). To carry out, the mix of the reaction was
used: a) 12.5 l iTaqTM Supermix with ROX (BioRad Laboratories, Hercules, CA,
USA) b) 0.25 l of each primers, c) 0.25 l tlh probe and 9,75l of ultrapure water.
The workings conditions are presenting in Table 1. Interpretation of the results
held in account the values of the cycle threshold (Ct) of the amplification curves.
The experiment was carried out in triple.
Table 1
Real-time PCR amplification conditions

N cycles Temperature Time


1 cycle Initiation 50C 2 min.
1 cycle Denaturation 95C 3 min.
Amplification of the t Denaturation 95C 15 sec.
of DNA segment (PCR Hybridisation 60C 60 sec.
40 cycles
cycling, collect and
data analysis) Extension 72 C 60 sec.

Efficiency. DNA from V.p. strain SR27 was prepared and decimal dilutions
were made. Real time PCR was performed using tlh primers and double dye
probe selected using the primer express software.
Specificity. The bacteria enumeration was made on TCBS in spiral (6).
From the pure culture one carried out a series of decimal dilutions, until dilution
10-7, numbered A0-A7, by using APW 3% salt medium. One carried out the
extraction of the ADN of each dilution, by respecting the protocol carefully
describes for the negative Gram bacteria. From this ADN one carried out a series
of numbered decimal dilutions V0-V7.

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Artificial contamination of mussels and shrimps. Twenty five grams of


mussels flesh were homogenised in 225 ml of alkaline buffered peptone with 3%
of NaCl. The homogenate was inoculated with serial dilutions of a fresh culture of
SR27 strain V.p.. The plates were incubated at 42C for 18 h. One ml of the
mixture was withdrawn and used to perform a DNA extraction and a real time
PCR experiment.

RESULTATS AND DISCUSSIONS

Specificity.To detect V.p. by PCR in real time, it was initially necessary to


choose a specific target gene, primers and a specific tlh probe (4). If the literature
is looked, at one note that several genes target are used to detect V.p. by PCR (3);
it acts of the genes toxR, orf8 and tlh (9). However, the gene orf8 is specific
serotype 03:K6 and the PCR of toxR is not positive for all the stocks of V.p.,
whereas all the stocks of V.p. have the gene tlh. Consequently, the tlh gene
seemed the target of choice to quickly detect V.p. (5).
Efficiency. The results of Ct obtained after the PCR in real time with the
TaqMan probe are presented in Fig. 1. The limit of detection was 1.2 copies,
corresponding to 40 CFUs/ml.
One noticed a difference of Ct between amplifications of the series coming
from dilution from the DNA (Fig. 3) from reference and the series coming from
bacterial dilutions which holds of the more important concentration in the sample
of ADN of reference.
For each series of bacterial dilutions and DNA dilutions, it obtained two
standard lines with almost identical slopes: for the series of bacterial dilutions
R2=0,988966 (Fig. 2) and for the series of dilutions of bacterial DNA
R2=0,993507 (Fig.4).

R2=0,98
Slope= -2,89
Efficiency= 121%

Fig. 2 Standard line:


Fig. 1 Amplifications curves: bacteria dilutions (A0 A7)
bacteria dilutions (A0 A7)

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R2=0,99
Slope= -3,29
Efficiency= 101%

Fig. 3 Amplifications curves: Fig. 4 Standard line:


DNA dilutions (V0 V7) DNA dilutions (V0 V7)

However, the extreme sensitivity of the real time PCR is not without
problem. Indeed, the least contamination of the PCR will be detected, it is thus
necessary to be particularly careful at the time of handling.
It is consequently recommended to use the system of the uracyl-n-
glycosylase to get rid of the products of preceding PCR which could come to
distort the results. Moreover, one must take care to the maximum to avoid the
contaminations of well. Moreover, one notes that the PCR in real time is a
technique more significant than the traditional PCR, since contaminations are
detected in PCR in real time but not in traditional PCR (7).
Artificial contamination of mussels and shrimps. The detection limit was
between 1 and 10 CFUs/25g. It was possible to detect V.p. in mussels and
shrimps. The limits of detection were: 12.6 CFUs/g of mussels and 40 CFUs/g of
shrimps.

CONCLUSIONS

It is possible to detect effective and fast V. parahaemolyticus in the fishery


products by the real time PCR with a TaqMan probe for tlh gene, and, in same
time, quantitatively to appreciate the load of this bacterium in the sample.
The fragment of the gene tlh, which is considered to mark it species by its
presence in the pathogenic stocks of V.p. and the environmental stocks,
nonpathogenic.
Although, no obvious inhibition of PCR was noted, it is recommended
when one works with complex matrices like the food matrices to use an internal
control of PCR.

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REFERENCES

1. AFNOR - Association Franaise De Normalisation. Norme franaise NF - ISO


8914:1990 (F): Directives gnrales pour la recherche de V. parahaemolyticus,
e
In : Association franaise de normalisation (Ed.), Microbiologie alimentaire, 8
dition, Tome 2 : Mthodes de routine et valuation dune mthode alternative,
Association franaise de normalisation, Paris, 2002, 147-163.
2. Applied Biosystems, Handbook for ABI PRISM 7000 Sequence Detection System,
adresse URL: http://www.applied-byosytems.com, consult le 12 octobre 2004 .
3. Blackstone G., Nordstrom J., Vickery M., DePaola A., 2003 - Detection of
pathogenic V. parahaemolyticus in oyster enrichments by real time PCR, J.
Microbiol Meth., 53 (2): 149-155.
4. Bradatan G., China B., Daube G., 2005 - Detection an characterisation of V.
parahaermolyticus in seafood using real time PCR. 10-th Conference on Food
Microbiology. Lige, 23-34 June 2005, poster.
5. CEE - Communaute Europeenne. Opinion of the Scientific Committee on Veterinary
Measures relating to Public Health on V. vulnificus and V. parahaemolyticus (in
row and undercooked seafood). 19-20 sept. 2001, http://www.europe.eu.int.
6. CEFAS - The Centre For Environment Fisheries & Aquaculture Science.
Enumeration of total and pathogenic V. parahaemolyticus, reference
SOPwa3.3, Weymouth Laboratory, Angleterre, 2004
7. China B., Ghafir Y., Daube G., 2002 - Estimation quantitative et qualitative par
amplification gntique des bactries prsentes dans les denres alimentaires,
Ann. Md. Vt, 147, 99-109.
8. Genbank : http://www.ncbi.nlm.nih.gov
9. ICMSF - International Commission On Microbiological Specifications For Food. V.
parahaemolyticus, In: Roberts T.A. (chairman Ed.), Microorganisms in foods,
e
5 : Characteristics of microbial pathogens, Blackie Academic Professional,
London, New York, Melbourne, Tokyo, 1996, 426-435.
10. Roberts T., 1996 - V. parahaemolyticus. In: Roberts T. et al. Microorganismes in
foods characteristics of microbial pathogens, Blackie Academic Professional,
London, 1996, 426-435.

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SENSIBILITATEA I SPECIFICITATEA UNOR TESTE


DE DIAGNOSTIC SEROLOGIC UTILIZATE
N TUBERCULOZA BOVIN
THE SENSIBILITY AND THE SENSITIVITY OF THE SEROLOGYCAL DIAGNOSIS
TESTS USED IN THE DIAGNOSIS OF THE BOVINE TUBERCULOSIS

Adriana CHIRIAC1, M. CARPCRARE2,


C. NECULI1
1
D.S.V.S.A. Vaslui
2
U.S.A.M.V. Iai

The necessity of introducing some complementary tests in


tuberculosis into practice, results from the fact that the tuberculin testing
doesn t allways track down the incipient forms of infection and doesn t
dissociate the infection from the evoluting tuberculosis.
In this meaning, in this investigation we describe three of the
serologycal diagnosis tests used in infection with Mycobacterium bovis:
enzyme - linked immunosorbent assay (EIAs - IFN ), the glutaraldehyde
test and the complement fixation test.
During 2000-2005, a number of 743 blood samples of the bovine
with intradermal reaction to the single test (IDR-TU) and the simultaneous
test (IDR-TCS) were processed within Sanitary Veterinary Laboratory
Vaslui by enzyme - linked immunosorbent assay ( EIAs - IFN ) test.
Afterwords, we have taken the glutaraldehyde test and the complement
fixation test on a number of 168 bovine (166 bovine with intradermal
reaction) .
As a result of these tests, that s how we have apreciatted the
sensibility and the sensitivity in this way: the enzyme - linked
immunosorbent assay have the sensibility 84,62 % and the sensitivity 80,9
%, the glutaraldehyde test have the sensibility 81,82 % and the sensitivity
50,00 % and the complement fixation test have the sensibility 75,00 % and
the sensitivity 69,23%.

Key words: tuberculosis, diagnosis, enzyme - linked immunosorbent assay,


the glutaraldehyde test, the complement fixation test, sensibility,
sensitivity

MATERIALE I METODE

Testul imunoenzimatic EIAs- IFN a fost folosit din necesitatea de a


demonstra c testele de depistare a tuberculozei ,,in vivo sunt mult mai rapide i
au o sensibilitate i o specificitate crescut fa de metodele clasice utilizate
curent.Studiul a fost efectuat pe efectivele de bovine n vrst de peste 6 luni
care au fost supuse testului intradermotuberculinic simplu (IDR-TU). Animalele

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reagente (pozitive i/ sau dubioase) la IDR- TU au fost recontrolate dup 15-30


de zile prin metoda imunoenzimatic de detecie a gamma-interferonului,
procedeul sandwich (EIAs- IFN), folosind trusa omologat sub denumirea de
,,BOVIGAm - kit- EIAs (4).De la aceste animale s-au recoltat probe de snge pe
anticoagulant (heparin), probele au fost apoi stimulate cu PPD bovin i PPD
aviar.Dup incubarea culturilor de snge integral la 370C, timp de 16-24 de ore,
s-a recoltat plasma.Pentru fiecare prob s-a luat n calcul un martor de control,
nestimulat. Limfocitele stimulate separat cu PPD bovin i PPD aviar au eliberat
gamma-interferonul, al crui nivel de detecie a fost apreciat spectrofotometric n
densitate optic( DO450 nm). Rezultatele obinute au fost trecute n tabelul 1.
Materialul cercetat a constat n 743 probe snge prelevate pe anticoagulant de
la bovine reagente la testele alergice (TU i TCS).
Testul glutaraldehidei reprezint o metod extrem de simpl, rapid i puin
costisitoare, care nu necesit examinarea ulterioar a animalelor, ca n cazul
testului alergic (1). Testul glutaraldehidei ar putea fi utilizat n paralel cu
tuberculinarea la animalele care reacioneaz pozitiv la testul alergic (2).
Tehnica de lucru: se recolteaz cte 2 ml snge de la fiecare animal, pe
anticoagulant (EDTA) n cantitate de 15 mg/ ml de snge.Se omogenizeaz
foarte bine.
Din probele recoltate, s-a luat cte 1 ml de snge care a fost amestecat n
proporii egale cu o soluie proaspt de glutaraldehid 1,25% n tuburi
Wassermann. Soluia de aldehid glutaric s-a obinut prin amestecarea a 20 ml
ap distilat cu 1 ml de aldehid glutaric 25%.
Probele au fost examinate din minut n minut pentru a observa apariia
gelifierii, timp de 15 minute.
Cercetrile s-au efectuat pe 168 probe de snge provenite de la bovine
reacionate pozitiv la tuberculinare (testul simplu i testul comparativ simultan).
Rezultatele obinute au fost sintetizate n tabelul 2.
Reacia de fixare a complementului se bazeaz pe proprietatea
complementului de a interaciona cu complexele antigen-anticorp (sistemul
bacterian). Cnd combinarea unui antigen cu anticorpul specific are loc n
prezena complementului, acesta se fixeaz pe complexele imune (6).
Serurile de examinat se dilueaz n sol. fiziologic n proporia 1/5 i apoi
se inactiveaz 30 de minute la 56C. Dup inactivare, se adaug antigenul diluat
conform titrului i calculat la volumul de 0,5 ml pentru fiecare tub.n aceast
reacie s-a folosit ca antigen tuberculin bovin (PPD bovin). Apoi se adaug
complementul, diluat n momentul folosirii n reacie, conform titrului stabilit n
titrarea preliminar i calculat la volumul de 0,5 ml pentru fiecare tub. Tuburile se
agit i apoi se introduc la baia de ap la 37C timp de 30 minute, pentru
incubarea reaciei.
Dup incubare, n fiecare tub se distribuie cte 1 ml din sistemul hemolitic
preparat n prealabil. Tuburile se agit din nou i se introduc la baia de ap la 37
timp de 15 minute, pentru incubarea reaciei, apoi se scot i rezultatele se
apreciaz i se interpreteaz imediat.
Citirea reaciei i interpretarea rezultatelor se face prin aprecierea
gradului de hemoliz, respectiv a lipsei hemolizei.

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Cercetrile s-au efectuat pe 168 probe de snge provenite de la bovine


reacionate pozitiv la tuberculinare (testul simplu i testul comparativ simultan).
Rezultatele obinute au fost sintetizate n tabelul 3.

REZULTATE I DISCUII

Testul imunoenzimatic EIAs- IFN


Analiza datelor din tabelul 1 relev c un procent de 5,25% dintre bovine
au prezentat reacii pozitive la testul EIAs- IFN. Peste 26,5% dintre ele au fost
ncadrate la categoria reactante aviare. Procentul relativ crescut de reacii pozitive
la PPD aviar sugereaz infecii cu micobacterii atipice, cele din complexul M.
avium sau intervenia altor ageni etiologici care pot determina alte afeciuni
(echinococoz, fascioloz, bruceloz, leucoz etc.) (3, 5, 7). Un numr de 8
animale au reacionat neconcludent (1, 08%).
Tabelul 1
Rezultate test EIAs- IFN

Nr. de bovine Reactant


Pozitiv Negativ Neconcludent
Anul testate aviar
S.C. G.P. S.C. G.P. S.C. G.P. S.C. G.P. S.C. G.P.
2000 89 310 2 5 45 70 42 235 - -
2001 - 188 - 4 - 18 - 166 - -
2002 78 - 14 - 32 - 28 - 4 -
2003 30 - 4 - 8 - 18 - - -
2004 29 - 6 - 15 - 4 - 4 -
2005 19 - 4 - 9 - 6 - - -
Total 245 498 30 9 109 88 98 401 8 -

Pe un numr de 168 de animale s-au aplicat testele de diagnostic a


tuberculozei pe animalele n via. Dintre acestea, 22 de animale au reacionat
pozitiv la testul EIAs- IFN (13,1%) i 144 au fost reactante aviare (85,71%).
Din cele 168 de bovine luate n studiu, 166 reagente la examenul alergic
(testul unic i testul comparativ simultan) i examinate prin testul imunoenzimatic
EIAs- IFN, testul glutaraldehidei i reacia de fixare a complementului, s-a
diagnosticat tuberculoz n urma diagnosticului complex de laborator la numai 11
animale (6,55%).
n contextul examenului complex de laborator, s-a apreciat sensibilitatea i
specificitatea acestui test n urma rezultatelor obinute.
Ss = a/ a+c% = 11/ 11 + 2 % = 84,62 %
Sp= d/ d+b% = 88 / 88 +13 % = 87,13 %
a = real pozitive; b = fals pozitive; c = fals negative; d = real negative;

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Testul glutaraldehidei
Studiul s-a efectuat pe cele 168 de bovine reagente la testul EIAs- IFN.
Analiza datelor din tabelul 2 relev c 37,5% din bovine au reacionat pozitiv la
test, iar 17,26% au fost dubioase.
Animalele reacionate pozitiv la tuberculinare au reacionat pozitiv i la
testul glutaraldehidei, doar c timpul de gelificare a sngelui a fost diferit. Testul
glutaraldehidei a fost dubios la un numr de 24 de bovine reacionate pozitiv la
tuberculinare.
S-a calculat sensibilitatea i specificitatea acestui test, lund n considerare
confirmarea sau infirmarea rezultatului testului n funcie de rezultatele
examenului complex de laborator.
Ss = a/ a+c% = 9/ 9+ 2 % = 81,82 %
Sp= d/ d+b% = 8 / 8 + 8 % = 50,00 %
a = real pozitive; b = fals pozitive; c = fals negative; d = real negative;
Tabelul 2
Rezultate test glutaraldehid

Nr. de bovine Dubios/


Pozitiv Negativ
Anul testate Neconcludent
S.C. G.P. S.C. G.P. S.C. G.P. S.C. G.P.
2000 27 44 - 14 6 5 21 25
2001 - 19 - 11 - 5 - 3
2002 32 - 10 - 4 - 18 -
2003 12 - 6 - 3 - 3 -
2004 13 - 10 - 2 - 1 -
2005 21 - 12 - 4 - 5 -
Total 105 63 38 25 19 10 48 28

Reacia de fixare a complementului


Studiul s-a efectuat pe cele 168 de bovine reagente la testul EIAs- IFN.
Analiza datelor din tabelul 3 demonstreaz c 7,14% din animale au reacionat
pozitiv, 45,24% au fost dubioase.
Din cele 76 animale reacionate dubios la RFC, 60 au fost pozitive la testul
glutaraldehidei .
Cele 12 animale reacionate pozitiv la RFC au reacionat pozitiv i la testul
glutaraldehidei, gelificnd sngele n primele 6 minute 7 cazuri, iar restul ntre 7
i 10 i mai mult de 10 minute .
n funcie de rezultatele examenului complex de laborator, s-au calculat
aceeai parametri (sensibilitate i specificitate):
Ss = a/ a+c% = 6 / 6 + 2 % = 75 %
Sp= d/ d+b% = 9/ 9 + 4 % = 69,23 %
a = real pozitive; b = fals pozitive; c = fals negative; d = real negative;

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Tabelul 3
Rezultate test R.F.C.

Nr. de bovine Dubios/


Pozitiv Negativ
Anul testate Neconcludent
S.C. G.P. S.C. G.P. S.C. G.P. S.C. G.P.
2000 27 44 - 1 7 17 20 26
2001 - 19 - 1 - 15 - 3
2002 32 - 4 - 7 - 21 -
2003 12 - 2 - 8 - 2 -
2004 13 - 1 - 11 - 1 -
2005 21 - 3 - 11 - 7 -
Total 105 63 10 2 44 32 51 29

Sensibilitatea i specificitatea testelor de diagnostic ,, in vivo n


ansamblu
Ss = a/ a+c% = 7/ 7+ 2 % = 77,77 %
Sp= d/ d+b% = 2/ 2+ 4 % = 33, 33 %
a = real pozitive; b = fals pozitive; c = fals negative; d = real negative;
n tabelul 4 s-a efectuat un studiu de comparare a sensibilitii i
specificitii testelor de diagnostic,,in vitro.
Tabelul 4
Compararea sensibilitii i specificitii fiecrui test de diagnostic

Testul Sensibilitate % Specificitate %


Testul alergic 78,57 99,53
EIAs- IFN 84,62 80,9
Testul glutaraldehidei 81,82 50,00
R.F.C. 75,00 69,23
Combinarea testelor 77,77 33,33

Aceast schem de diagnostic ar putea fi utilizat n campaniile de


eradicare a tuberculozei cnd maximum de sensibilitate este necesar i o
reducere a specificitii poate fi tolerat.
n lucrarea de fa s-a demonstrat c testele celulare, respectiv EIAs- IFN,
au o sensibilitate i specificitate mult mai mare dect testele de detectare a
anticorpilor (R.F.C.), lucru susinut i de literatura de specialitate (8).

CONCLUZII

Testul EIAs- IFN, procedeul sandwich, este o metod expeditiv fa de


testul alergic (IDR- TU), rezultatele obinndu-se n 24 - 48 de ore, fa de 72 de
ore, n testele clasice.

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Testul EIAs- IFN poate fi repetat ori de cte ori este necesar, fr a
influena statusul imunitar al animalului, spre deosebire de testul alergic care
necesit un interval cuprins ntre 45 - 60, chiar 90 de zile, pentru a fi repetat.
Fiind un test biologic, EIAs- IFN poate furniza i unele rezultate fals-
pozitive i deci trebuie interpretat n contextul datelor epidemiologice i n
completarea testelor ,, in vivo.
Testul EIAs- IFN detecteaz infecia tuberculoas recent la bovine.
Toate animalele reacionate pozitiv la tuberculinare (testul unic i simultan)
i la reacia de fixare a complementului au reacionat pozitiv i la testul
glutarlaldehidei.
Se observ c prin combinarea testelor de diagnostic se obine o
sensibilitate ridicat (77.77%), prin comparaie cu o specificitate redus (33,33%).
Coroborarea rezultatelor testelor alergice cu rezultatele testelor
complementare tuberculinrii ( testul EIAs- IFN, testul glutaraldehidei, reacia
de fixare a complementului) poate constitui un suport mai raional i mai fidel
n ceea ce privete decizia de a ndruma la sacrificarea n abator a unui animal
reagent la tuberculin, dect testele alergice privite n mod singular.

BIBLIOGRAFIE

1. Carp-Crare M., Eleonora Guguianu, Marcel M., Eugenia Burtan, Dorina


Timofte, Moroan ., 1997 - Evaluarea preliminar a unei metode simple pentru
diagnosticul tuberculozei bovine prin testul glutaraldehidei, Universitatea
Agronomic i de Medicin Veterinar Iai, Lucrri tiinifice, vol.40, pag.164-
167.
2. Larsson S., 1988 A preliminary study an inexpensive method for the detection of
tuberculosis in cattle using the glutaraldehyde test, J.Nepal Med.Assoc.26, 9-17.
3. Miciora R., 1996 - Surse de eroare n diagnosticul alergic al tuberculozei bovine,
Revista Romn de Medicin Veterinar, vol.6, nr.1, 39-47.
4. Ni C., 2005 - Reacii specifice i nespecifice n diagnosticul tuberculozei bovine,
Revista Romn de Medicin Veterinar, vol.15, nr.2, 91-97.
5. Popescu t., Sandu I., 1997 - Preocupri i perspective n domeniul diagnosticului,
prevenirii i combaterii tuberculozei bovine - Ed. L.C.S.V.D. Bucureti, pag. 5-33.
6. Rducnescu H., Bica-Popii Valeria, 1986 Bacteriologie veterinar, Editura
Ceres, Bucureti
7. Rothell J.S., Jones S.L., Corner L.A., Wood P.R., 1992 - The gamma- interferon
assay for diagnosis of bovine tuberculosis in cattle; condition affecting the
production gamma-interferon in whole blood culture, Australian Veterinary
Journal, 69, 1-4.
8. Wood P.R., Rothel J.S., Corner L.A., Baldock C., Jones S.L., Mc Cormick B.,
Francis B.R., Creeper J., 1991 - Field compararison of the interferon - gamma
assay and the intradermal tuberculin test for diagnosis of bovine tuberculosis,
Vet.Microbio., 31, 71-79.

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INVESTIGAII PRIVIND CALITATEA MICROBIOLOGIC


A LAPTELUI COMERCIALIZAT DE PRODUCTORII
PARTICULARI
INVESTIGATIONS REGARDING THE MICROBIOLOGICAL QUALITY
OF THE MILK SOLD BY THE PRIVAT PRODUCERS

Eleonora GUGUIANU, Ana-Maria BLTESCU


U.S.A.M.V. Iai

In conformity with the actual legislation, the microbiological


investigation of milk is obligatory only in milk industry. In this context, the
fresh milk sold at market by peasantes is not investigated for sanitation. In
this work, we present the quantitative microbiological exams results
(N.T.G./ml and reductasis test) from 40 milk samples.
N.T.G.MA./ml oscilated between 50000 and 5350000 germs/ml,
corelated with reductasis test results.
Most of the samples were corect with the internel microbiological
normes (M.H. Order 975/1999),but there were incorect with the A.W.F.M
Order 389/2002 imposed by the necessity of armonisation with European
Community.

Conform legislaiei n vigoare, n ara noastr examenul microbiologic al


laptelui se efectueaz obligatoriu doar n unitile de prelucrare.
Deoarece, n acest context legislativ, laptele crud comercializat n piee de
ctre productorii particulari nu este supus unor investigaii de laborator privind
salubritatea, ne-am propus s testm gradul de poluare bacteriologic a acestui
sortiment de lapte prin determinarea numrului total de germeni mezofili aerobi
(NTGMA/ml) i proba reductazei.

MATERIAL I METAD

Investigaiile au fost efectuate pe un numr de 40 probe de lapte din sector


particular, recoltate din pieele agroalimentare ale oraului Iai, n perioada iunie-
iulie 2005.
Laptele, recoltat n eprubete sterile, a fost supus examenului microbiologic
dup aproximativ o or de la recoltare, n laboratorul de microbiologie alimentar
al Facultii de Medicin Veterinar Iai.
Aprecierea ncrcturii bacteriene a laptelui s-a efectuat prin determinarea
NTGMA/ml i proba reductazei metod indirect, simpl i expeditiv (2, 3).
NTGMA/ml a fost determinat prin metoda culturilor n plci pentru
determinarea unitilor formatoare de colonii la temperatura de 300C iar pentru
proba reductazei s-a folosit ca reactiv soluia de albastru de metilen 5% (5 ml
soluie alcoolic saturat de albastru de metilen i 195 ml ap distilat steril).

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Rezultatele obinute au fost raportate la normele microbiologice ale


ordinului Ministerului Sntii 975/1999 si ale ordinului Ministerului Agriculturii,
Alimentaiei i Pdurilor 389/2002, armonizate cu cele ale Uniunii Europene, care
se vor aplica ncepnd cu anul 2011.

REZULTATE I DISCUII

Examenele efectuate au evideniat niveluri variate de contaminare


bacterian a celor 40 de probe de lapte (tabelul 1).
Tabelul 1
Rezultatele testelor de apreciere a calitii microbiologice a laptelui

Proba Proba reductazei


NTGMA/ml
Nr. Timp Calitatea laptelui
1 780000 3h satisfctoare
2 50000 9h 25min buna
3 185000 7h 40 min buna
4 84000 8h 45 min buna
5 350000 5h 45 min buna
6 5350000 25 min nesatisfacatoare
7 630000 3h 45 min satisfctoare
8 825000 2h 15 min satisfctoare
9 600000 3h 30 min satisfctoare
10 740000 3h 30 min satisfctoare
11 560300 4h35min satisfctoare
12 120450 8h 20 min buna
13 68500 8h 30min buna
14 220000 7h 30 min buna
15 783500 3h 15 min satisfctoare
16 595000 4h 10 min satisfctoare
17 85300 8h 40 min buna
18 975000 2h 45 min satisfctoare
19 630000 3h 30 min satisfctoare
20 550000 4h 25 min satisfctoare
21 275000 6h 20 min buna
22 125000 7h buna
23 370000 6h 10 min buna
24 590000 5h satisfctoare
25 325000 6h buna
26 220800 7h 20 min buna
27 925000 2h 45 min satisfctoare
28 700000 3h 15 min satisfctoare
29 250200 7h 20 min buna
30 100000 8h 10 min buna
31 935000 2h 55min satisfctoare
32 970500 2h 30 min satisfctoare

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Proba Proba reductazei


NTGMA/ml
Nr. Timp Calitatea laptelui
33 945000 2h 45 min satisfctoare
34 820000 2h 15 min satisfctoare
35 970000 2h 45 min satisfctoare
36 620000 3h 30 min satisfctoare
37 500000 4h 45 min satisfctoare
38 695000 3h 15 min satisfctoare
39 580000 4h 05 min satisfctoare
40 730000 3h 10 min satisfctoare

Analiznd datele din tabel prin raportarea lor la normele microbiologice


prevzute de legislaia n vigoare rezult c din cele 40 de probe:
- 39 probe (97,5%) au o ncrctur bacterian situat sub limita
maxim admis pentru laptele crud integral ( 1000000 germeni/ml);
- 1 prob - proba nr. 6 - (2,5%) depeste limita maxim admis pentru
laptele crud integral;
Rezultatele obinute la testul reductazei concorda cu N.T.G. si indic doua
categorii de calitate:
- lapte bun (sub 500000 germeni/ml) 15 probe (37,5%);
- lapte satisfctor (500000-4000000 germeni/ml) - 25 probe (62,5%);
Calitatea ,,lapte satisfacator din punct de vedere microbiologic permite o
incarcatura microbiana de 4 ori mai mare comparativ cu valoarea maxima admisa
a indicatorul NTG/ml, incluzand proba de lapte nr.6.
In raport cu cerintele ordinului M.A.A.P 389/2002 (N.T.G. /ml lapte crud
<= 1000000), doar 5 din cele 40 de probe investigate - probele nr. 2,4,13,17 si 30
- (12,5%) sunt salubre din punct de vedere microbiologic.

CONCLUZII

S-au efectuat examene microbiologice cantitative (NTGMA/ml i proba


reductazei) pe 40 probe de lapte crud, comercializat in piata de producatori
particulari;
NTGMA/ml a oscilat intre 50000 si 5350000 germeni/ml lapte;
Majoritatea probelor (97,5%) s-a incadrat in normele microbiologice
interne ale legislatiei in vigoare ( Ordinul M.S. 975/1999);
Calitatea microbiologica a laptelui a fost necorespunzatoare la 87,5% din
probe in raport cu exigentele ordinului M.A.A.P 389/2002 (87,5%).

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BIBLIOGRAFIE

1. Brzoi D., Apostu S., 2002 Microbiologia produselor alimentare. Ed. RISOPRINT,
Cluj-Napoca.
2. Bondoc I., indilar E.V., 2002 - Controlul sanitar veterinar al calitii i salubritii
alimentelor. Ed. Ion Ionescu de la Brad. Iai.
3. Sindilar E., 2000 Controlul igienic al produselor i subproduselor de origine
animal. Vol.1, 2. Ed. I.N.R.C.S. Iasi.
4. *** Ordinul M.S. 975/1998 privind aprobarea Normelor igienico-sanitare pentru
alimente.
5. *** Ordinul M.A.A.P. 389/2002 pentru aprobarea Normei sanitare veterinare privind
conditiile de sanatate pentru producerea si comercializarea laptelui crud, a
laptelui tratat termic si a produselor pe baza de lapte.

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EFFICACY OF SAME ANTIMICROBIAL SUBSTANCE


AT PATHOGEN MICROORGANISMS ISOLATED
IN POULTRY BREEDING OBJECTS
. ILI1, LJ. SPALEVI1, B. MILJKOVI1,
I. PAVLOVI1, G. UGI2, S. STANOJEVI1
1
Scientific Veterinary Institute of Serbia
2
Medicines and Medical Devices Agency of Serbia

Diseases of bacterial ethiology presenc important factors in poultry


production, therefor source to spread infection in poultry flocks and
eventual econimc losess which they induced. Our examination was aimed to
determinate main bacterial contaminant at breeding objects at poultry
farms and influence of several most utilizes disinfectant to isolated
pathogen microorganism. During examination we occurred presence of
Salmonellae spp., E.coli, Staphylococcus spp. and Aspergillus spp. To our
examination we tested antibacterial efficacy of next disinfectant: phenol,
potassium hydroxide, potassium dichloride izocyanurat, perchlor vinegar
acid, iodophor, kaliumperoxisulfat, amphotenzide, and formaldehyde.
Formaldehyde we outstanding like disinfectant with best antibacterial
action because they destroyed tested microorganism in both using
concentration promptly after exposure. Perchlor vinegar acid and iodophor
was efficiency after exposition of 5 minutes in both concentrations to all of
tested microorganisms. Kaliumperoxisulfate, phenol and amphotenzide
were efficiency in all concentration after 10 minutes to all tested
microorganisms. Potassium dichloride izocyanurat was show efficacy
against all microorganisms after 24 hours exposition.

Key words: Salmonella spp., E. coli, Staphylococcus spp., Aspergillus spp.,


disinfectants, poultry farm

INTRODUCTION

Diseases of bacterial etiology presence important factors in poultry


production, therefore source to spread infection in poultry flocks and eventual
economic losses, which they induced. Great agglomeration of poultry at object
induced increased of pathogenesis of same microbial agents, especial bacterial,
which induced infection with high rate of morbidity and mortality (Langsrud and
Sundheim 1998., Hristov et al.. 1996). Zoonotic potential of same microbial
agents like Salmonella spp, presence a special epidemiological problem in poultry
breeding (Pavlovi et al.1988.,Mayhall 1996).
From these reason in mind we performed our examination, which was
aimed to determinate main bacterial contaminant at breeding objects at poultry

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farms, and influence of several most utilizes disinfectant to isolated pathogen


microorganism.

MATERIAL AND METHODS

During the period 2000-2005 we performed an examination of 24 poultry


farms. Examination of microorganisms species in object was performed using
swabs at various parts of object (floor, walls, etc.), which examined with routine
microbiological laboratory methods for bacteriology cultivation and determination
at laboratory of Dep.of Poultry Diseases in Scientific Veterinary Institute of Serbia
To our examination we tested antibacterial efficacy of next disinfectant:
phenol (1 and 2% concentration), potassium hydroxide (2 and 3%), potassium
dichloride izocyanurat (0.02 and 0.04%), perchlor vinegar acid (2 and 3%),
iodophor (0.1 and 0.3%), kaliumperoxisulfate (1 and 2%), amphotenzide (1 and
2%), and formaldehyde (5 and 10%). Survive of found bacterial strain were
examined 1, 5 and 10 minutes and 24 hours after exposition to each of
disinfectant. That control we performed using swabs at various parts of objects
and its microbiological examination by method described by Cremieux and
Fleurette (1991).

RESULTS AND DISCUSION

During examination we detected the presence of Salmonellae spp., E.coli,


Staphylococcus spp., and Aspergillus spp.
During our examination, formaldehyde we outstanding like disinfectant
with best antibacterial action because they destroyed tested bacterial
microorganism in both using concentration promptly after exposure. Followed by
perchlor vinegar acid and iodophor was efficiency after exposition of 5 minutes in
both concentrations to all of tested microorganisms. Kaliumperoxisulfate, phenol
and amphotenzide were efficiency in all concentration after 10 minutes to all
tested microorganisms. Potassium dichloride izocyanurat was show efficacy
against all bacterial microorganisms after 24 hours exposition.
With fungal agents best result showed formaldehyde, because they
destroyed tested fungal microorganism promptly after exposure; followed by
perchlor vinegar acid, iodophor and amphotenzide which destroyed it at first 1
minute of application. Kaliumperoxisulfate and phenol were efficiency in all
concentration after 5 minutes. Potassium dichloride izocyanurat was show
efficacy after 10 minutes.
Those results especially at results of formaldehyde efficacy are similar like
results of examination performed by Tucker et al. (1975): Andryunin (1982), and
Dietz et al (1980). Efficacy of other disinfectants was analogous with results of
Bodiroga et al. (1996), Mijatov et al. (1998) and Ili (2004).

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CONCLUSION

Results of our examination shown that formaldehyde we outstanding like


disinfectant with best antibacterial and antifungal action because they destroyed
tested microorganism in both using concentration promptly after exposure. Other
disificiant present minimum wariation at exposition time and efficacy against
bacterial and fungal microorganisms.

REFERENCE

1. Andryunin Y.I., 1982 - Theoretical basis of the bactericidal activity of an alkaline


solution of formaldehyde. Vestnik Selskokhozyaistvennoi Nauki, USSR,9, 80-
84.
2. Bodiroga Jovanka, Viakci M. Aanin Ruica, Pisarev Mira, 1996 - Nove
mogunosti dezinfekcije vode za pie, prostora i opreme preparatom Peral-S
na ivinarskim farmama. VII savetovanje DDDD u zatiti ivotne sredine,
Sutomore, 32-33.
3. Cremieux A., J. Fleurette, 1991 - Methods of testing disinfectans u Block, S.S.:
Disinfection, sterilization and preservation. Philadelphia, Lea Febiger (4
izdanje) 1009-1027.
4. Dietz P., Bohm R., Strauch D., 1980 - Effectivenss and safty of formaldehyde gas
and aerosol of peracetic acid and hydrogen peroxide. Zentralblatt fr
Veterinarmedizin, German federal Republic, 27B(4), 268-279,
5. Hristov S., Anoji B., Vuini Marija, Stankovi B., 1996 - Uticaj praine vazduha
na organizam ivotinja. Zbornik radova,VII savetovanje DDDDD u zatiti
ivotne sredine, Sutomore, 268-275.
6. Ili M., Damnjanovi N., Plea M., 1993 - Ispitivanje Galisepta u laboratorijskim i
praktinim uslovima. IV simpozijum DDD u zatiti ivotne sredine, Vrnjaka
Banja, 27-30.
7. Ili ., 2004 - Influence of antimicrobial substance at pathogen bacterial strains and
fungal in objects of poultry breeding, MsC thesis, faculty of Veterinary Medicine
in Belgrade
8. Langsrud S., Sundheim G., 1998 - Factory influencing a suspension test method for
microbical activity of disinfectans. Journal of Appl. Mycrobiology, 1006-1012.
9. Mayhall C.G., 1996 - Hospital epidemiology and infection control. London: Willians
and Wilkins, 927.
10. Mijatov Lj., Sajti G., Grbovi Mirjana, Molnar-Vojni Danica, 1997 - Prednosti i
preimustva DESU amfotenzida u praksi. VIII savetovanje DDDDD u zatiti
ivotne sredine.Zbornik radova, Subotica, 39-40.
11. Mijatov Lj., Mirjana Grbovi, Danica Molnar, 1998 - Dejstvo DESU amfotenzida na
animalne patogene bakterije. Zbornik radova, IX savetovanja DDD.
12. Pavlovi I., Blain Vesna, Ili ivka, 1988 - Salmoneloza ivine tokom 1987.godine
na uoj teritoriji SR Srbije i njihovo suzbijanje Zbornik radova ivinarski dani
Jugoslavije, Pritina, 133-144;
13. Payne D.N., Babb J.R., Bradlly Cr., 1999 - Anerotnation of the smitability of the
European suspension test to reflect in vitro activity of antiseptics against
clinically significant organisms. Letters in Appl. Mycrobiology, 7-12.
14. Tucker J.F., Harry E.G., Wainman H.E., 1975 - The effect of fumigation with methyl
bromide or formaldehyde on the infectivity of poultry house litter naturrally
contaminated with Salmonella virchow. British Veterinary Journal, UK, 131(4),
474-485.

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EVALUAREA IZOLRII SPECIILOR VIBRIO NON-


HOLERICE DIN PROBE DE AP PRELEVATE DIN SUDUL
I SUD-ESTUL RII
THE ISOLATION EVALUATION OF NON-CHOLERIC VIBRIO SPECIES
IN SOUTH AND SOUTHEAST ROMANIAN WATERS

L. TUDOR, I. OGOE
U.S.A.M.V. Bucureti

The investigations have searched the harvesting frequency of


bacterial species of non-choleric Vibrio from aquatic samples. There were
harvested substantiations from fresh waters and salted waters. These
researches are continuing the last years began studies, for realize a
completed and detailed statistics.
Between 1997-2005 have been prelevated and prepared a total
number of 4585 water substantiations for south and southeast Romanian
waters.
From the total number of analyzed substantiations were isolated 161
stems of non-choleric Vibrio species representing a total harvesting
frequency of 3.51 %. The annual harvesting frequency has been: in 1997 of
4,05%; in 1998 of 3,92%; in 1999 of 4,237; in 2000 3,26%; in 2001 of 4,18;
in 2002 of 4,36%; in 2003 of 2,51%; in 2004 of 2.03% and in 2005 of
3.08%.

Key-words: incidence, vibrions, non-choleric, freshwater, saltwater

Speciile bacteriene ncadrate n genul Vibrio care au semnificaie clinic


(sunt patogene pentru om sau diferite specii de animale) sunt de obicei prezente n
mediul exterior i formeaz o microflor autohton a apelor din bli, lacuri
srate, estuare, mri i oceane din vecintatea rmurilor situate n zonele
temperate i calde (3, 14). Unii specialiti consider c sunt specii ubiquitare,
deoarece supravieuiesc i se multiplic n cele mai variate medii sau n produsele
alimentare (3, 6, 10).
Dei pe plan mondial s-au fcut numeroase acumulri tiinifice privind
toxiinfeciile alimentare pe care le produc la om unele bacterii ale genului Vibrio,
n ara noastr exist puine date referitoare la incidena toxiinfeciilor alimentare
produse de vibrionii non-holerici (Vibrio vulnificus, Vibrio fluvialis, Vibrio
mimicus, Vibrio alginolyticus, Vibrio metschinikovii, Vibrio parahaemolyticus,
Vibrio furnissii, etc) i mai ales studii referitoare la prezena acestor bacterii n
ape, pe suprafaa corpului sau n cavitile naturale ale diferitelor organisme
acvatice, precum i n diferite tipuri de alimente. Aceste motive relev
oportunitatea investigaiilor referitoare la prevalena speciilor bacteriene ncadrate

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n genul Vibrio, din diferite produse alimentare cu origine acvatic: pete,


crustacee, molute, etc.
Mai muli autori au prezentat studii ample privind izolarea vibrionilor non-
holerici din diferite tipuri de medii (ape dulci, ape srate, sedimente) constatnd
totodat capacitatea deosebit a acestor bacterii de a se adapta i multiplica (2, 3,
6, 8, 11, 14). La nivel animal, vibrionii au fost izolai din zooplancton, molute
(lamelibranhiate, cefalopode sau gasteropode), din organele interne i musculatura
diferitelor crustacee acvatice, din intestinul i ficatul petilor, din musculatura i
diferitele organe interne ale mamiferelor acvatice (2, 3, 6, 8, 11, 14).
n ultimii ani s-au evideniat probe despre existena unor rezervoare de
vibrioni n diferite tipuri de medii naturale (sedimente, plancton), bacteriile fiind
preluate i concentrate n organism de ctre animalele acvatice. Studiile fcute n
apele de coast ale SUA i Australiei au demonstrat existena unor rezervoare de
vibrioni non-holerici n crustaceii i molutele cu cochilie. De aceea n unele ri,
cum ar fi SUA, sursa principal de infecie pentru om o reprezint consumul
stridiilor, iar unele episoade de toxiinfecii alimentare apar n urma consumrii de
alimente cu origine marin, n principal carne netratat termic provenit de la
specii acvatice.
Prevalena toxiinfeciilor alimentare produse de diferitele specii Vibrio
non-holerice poate s ating n unele ri (cum ar fi Japonia) valori de peste 70%
din totalul cazurilor de infecii cu etiologie bacterian. Numrul de cazuri din
fiecare episod variaz de la o persoan la mai multe sute de persoane. Aceste
studii efectuate n alte ri au impulsionat demararea cercetrilor noastre, pentru a
constata existena vibrionilor n apele salmastre din Romnia, frecvena de izolare
din diferite tipuri de medii naturale i probe biologice. Un alt motiv pentru care
am considerat necesar studierea vibrionilor non-holerici n unele biotopuri a fost
determinat de necesitatea cunoaterii modului n care aceste bacterii circul n
ecosistemele acvatice.

MATERIAL I METODE

Au fost prelevate probe de ap din reeaua de lacuri a zonei de sud i sud-


est, din sistemul Razelm - Sinoe, zona Delta Dunrii i din zona litoral a Mrii
Negre.
Recoltrile au fost fcute pe o perioad de 9 ani, anual examinnd un
numr reprezentativ de probe (tabelul 1).
Probele recoltate au fost prelucrate prin metode speciale pentru izolarea i
identificarea speciilor bacteriene ncadrate n genul Vibrio, utiliznd comparativ
diferite metode (STAS ISO 8914, metodele FAO-OMS, metodele sintetice James
D. Oliver, metodele originale).

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Tabelul 1
Probele de ap recoltate i prelucrate n vederea identificrii
vibrionilor non-holerici

Zona de recoltare
Perioada Lacuri din Complex Zona litoral
Zona Delta Total
zona de sud Razelm- Marea
Dunrii probe
i sud-est Sinoe Neagr
1997 116 78 124 127 445
1998 124 67 115 179 485
1999 107 92 139 211 549
2000 115 81 127 198 521
2001 98 86 117 154 455
2002 112 102 132 204 550
2003 121 95 121 182 519
2004 127 98 116 201 542
2005 1116 97 119 187 519
Total
1036 796 1110 1643 4585
perioad

Probele de ap transportate n recipiente sterile i ermetic nchise sau n


medii de transport, n laborator au fost introduse n cte dou tipuri de medii de
mbogire. O parte a probei a fost introdus n cte 250 ml ap peptonat salin
alcalin, apoi incubat timp de 20 24 de ore la 370C; alt parte din prob a fost
introdus n cte 250 ml bulion salin cu polimixin B, apoi incubat la 35 370C
timp de 7 8 ore.
Dup incubare, din cultura total obinut, au fost striate cte 2 tipuri de
medii solide de izolare selectiv (de obicei au fost utilizate geloza tiosulfat-citrat-
bil-zaharoz TCBS i geloza tripsin-pepton-soia-clorur de
trifeniltetrazolium TSAT). Incubarea s-a realizat timp de 18 24 ore, la 35
370C.
Din culturile caracteristice obinute pe mediile de izolare selectiv (colonii
netede galbene mari sau colonii netede verzui cu centrul albastru-verzui pe
TCBS, sau colonii rou-viinii pn la rou-brun pe mediul TSAT) au fost preluate
fragmente care s-au transplantat pe agar nutritiv salin i n bulion nutritiv salin;
subculturile obinute, prin incubare 18 24 ore la 35 370C, au fost utilizate n
etapa de identificare.
Identificarea s-a efectuat fie pe baterie de teste biochimice, fie pe galerii de
identificare Api (Api Campy sau Api 20 E).

REZULTATE I DISCUII

Dintr-un numr total de 4066 probe de ap au fost izolate 145 tulpini


bacteriene aparinnd genului Vibrio.
Rezultatele obinute sunt redate sintetic n tabelul 2.

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Tabelul 2
Frecvena de izolare a vibrionilor non-holerici din probe de ap

Zona de recoltare
Lacuri din Zona
Perioada Complex Zona
zona de litoral Total
Razelm- Delta
sud i Marea probe
Sinoe Dunrii
sud-est Neagr
Nr. tulpini 0 1 7 10 18
1997
% 0 1,28 5,65 7,87 4,05
Nr. tulpini 1 0 6 12 19
1998
% 0,81 0 5,22 6,70 3,92
Nr. tulpini 0 1 8 15 24
1999
% 0 1,09 5,76 7,11 4,37
Nr. tulpini 0 1 5 11 17
2000
% 0 1,23 3,94 5,56 3,26
Nr. tulpini 1 2 4 12 19
2001
% 1,02 2,33 3,42 7,79 4,18
Nr. tulpini 1 2 7 14 24
2002
% 0,89 1,96 5,30 6,86 4,36
Nr. tulpini 0 0 6 7 13
2003
% 0 0 4,96 3,85 2,51
Nr. tulpini 1 1 3 6 11
2004
% 0,79 1,02 2,59 2,99 2,03
Nr. tulpini 1 1 5 9 16
2005
% 0,86 1,03 4,20 4,81 3,08
Total Nr. tulpini 5 9 51 96 161
perioad % 0,48 1,13 4,59 5,84 3,51

Analiza rezultatelor obinute demonstreaz o prevalen total pentru


perioada luat n studiu de 3,51%.
n cadrul zonelor analizate, incidena cea mai ridicat a fost constatat n
zona litoralului Mrii Negre (5,97%), iar cea mai sczut inciden a fost gsit n
zona lacurilor din sud i sud-est (0,43%).

CONCLUZII

Prevalena total a izolrii de bacterii din genul Vibrio (vibrioni non-


holerici) pentru perioada luat n studiu a fost de 3,51. Variaiile anuale s-au situat
n limite strnse, ntre un maximum de 4,37% i un minimum de 2,03%.
Prin analiza statistic a datelor obinute s-a constatat c frecvena de izolare
a vibrionilor non-holerici este mai mare pentru probele prelevate din zona litoral
(cu un maxim de 7,79% n anul 2001) comparativ cu zonele fluviale (cu un
maxim de 1,02 % n anul 2001).
Frecvena de izolare a vibrionilor non-holerici din probele de ap cunoate
variaii anuale care dovedesc o ciclicitate (perioade de maximum nregistrate n

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toate apele i perioade de declin), pn n prezent putndu-se constata o


periodicitate de cretere a incidenei de izolare de cca. 3 ani.

BIBLIOGRAFIE

1. Cerda-Cuellar M., Jofre J., Blanch A.R., 2000 A selective medium and a specific
probe for detection of Vibrio vulnificus. Appl & Environ. Microbiol. 66 (2): 855-9
2. Oliver J.D., 1989 Vibrio vulnificus. In M. P. Doyle Bacterial pathogens in foods.
Ed. M. Dekker, New York
3. Oliver J.D., Kaper J.B., 1997 Vibrio species, cap 13, in M. P. Doyle Bacterial
pathogens in foods. Ed. M. Dekker, New York
4. Rpuntean Gh., Marica D., Pop M., Manolescu N., Rudreanu Natalia, 1993 An
Original Soft-Agar for Selective Isolation of Vibrio cholerae and Vibrio furnissi
from Waters, Foods and Pathological Materials. Simpozion WAVFH, Bangkok,
Thailanda, 514-515
5. Tudor L., ogoe I., Stnescu V., 2001 Schem simplificat pentru izolarea i
identificarea unor specii aparinnd genului Vibrio. Al IV-lea Simpozion
aniversar al Institutului de Diagnostic i Sntate Animal 35 de ani de
activitate n diagnosticul medical veterinar pentru aprarea sntii animale i
umane, Bucureti, 145
6. Tudor L., Stnescu V., ogoe I., 2001 Prevalena speciilor patogene de Vibrio n
ape, sedimente, animale acvatice i unele produse de origine animal. Al IV-lea
Simpozion aniversar al Institutului de Diagnostic i Sntate Animal 35 de ani
de activitate n diagnosticul medical veterinar pentru aprarea sntii animale
i umane, Bucureti, 144.
7. Tudor L., ogoe I., Stnescu V., 2001 Frecvena izolrii unor specii de Vibrio
patogene pentru om, n zona de sud i sud-est a rii. Al IV-lea Simpozion
aniversar al Institutului de Diagnostic i Sntate Animal 35 de ani de
activitate n diagnosticul medical veterinar pentru aprarea sntii animale i
umane, Bucureti, 146
8. Tudor L., 2001 Prevalena bacteriilor din genul Vibrio n alimente de origine
animal, potenial implicate n toxiinfecii alimentare la om i nsuirile
morfobiologice ale acestora. Tez de doctorat, FMV Bucureti
9. Tudor L., ogoe I., 2001 Un nou mediu selectiv pentru izolarea speciilor non-
holerice din genul Vibrio. Simpozion omagial 140 de ani de nvmnt
medical veterinar romnesc, FMV Bucureti, 45
10. Tudor L., 2001 Evaluarea contaminrii produselor alimentare de origine acvatic
cu specii Vibrio non-holerice. Simpozion omagial 140 de ani de nvmnt
medical veterinar romnesc, FMV Bucureti, 135
11. Tudor L., 2002 Morfologia, biologia i implicaiile pentru patologie ale vibrionilor
non-holerici. Ed. Printech, Bucureti
12. Tudor L., Mitrnescu Elena, ogoe I., 2003 Evaluarea contaminrii cu vibrioni
non-holerici a petilor capturat din apele zonei de sud i sud-est a rii. Lucr. t.
Med. Vet. Timioara, Vol. XXXVI
13. Tudor L., Mitrnescu Elena, ogoe I., 2003 Prevalena izolrii speciilor Vibrio
non-holerice la molute acvatice capturate sau comercializate n zona de sud i
sud-est a rii. Rev. Rom. Med. Vet., Bucureti, vol. 13, p. 114
14. Twed R.T., 1989 Vibrio parahaemolyticus. In M.P.Doyle Bacterial pathogens in
foods, Ed. M. Dekker, New York, p. 569

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EVALUAREA CALITII MICROBIOLOGICE A UNOR


BRNZETURI COMERCIALIZATE PE PIEELE
BUCURETENE
THE EVALUATION OF MICROBIOLOGICAL QUALITY FOR SOME TYPES
OF CHEESE TRADED ON BUCHAREST MARKETS

L. TUDOR, I. OGOE,
Elena MITRNESCU
U.S.A.M.V. Bucureti

The researches have been effectuated for determining the


contamination grade of market cheese, following the supposition that all
products are delivered on right analysis documents. It was analyzed a total
number of 954 cheese samples. The samples have been harvested from 9
kinds of cheese from different market places.
The obtained statistical analyzed results have been permitted the
conclusion that entire types of cheese have been recorded unconformity
samples. It have been remarked either the getting over of some microbial
parameters (coliformes, coagulase-positive staphylococci, sulfite-reducing
bacteria) or the presence of some pathogen and pathogen-conditioned
bacteria (Clostridium perfringens has been isolated from 1.68% samples,
Proteus sp. has been isolated from 0.84% samples).

Key words: cheese, evaluation, quality, bacteria, contamination

Calitatea microbiologic a brnzeturilor reprezint un domeniu al siguranei


alimentare, deoarece prin prezena n produsul alimentar a speciilor bacteriene
patogene sau condiionat patogene, sau datorit degradrii substratului nutritiv i
formrii unor compui toxici, pot fi generate o serie de entiti morbide
consumatorilor [2, 4, 5].
Avnd n vedere aceste considerente, n ultimii ani au fost derulate
programe ample de modernizare a unitilor de procesare a laptelui i au fost
achiziionate utilaje performante care asigur scderea numrului de contaminani
n produsul finit, n acelai scop fiind preluate i adaptate noi tehnologii i fluxuri
de procesare, mai eficiente i care corespund cerinelor actuale de igien n
procesare [5, 8].
Efectul direct al acestor msuri s-a reflectat n primul rnd n calitatea
produselor lactate n general, mbuntirea parametrilor microbiologici i fizico-
chimici ai semifabricatelor i produselor finite analizate imediat dup procesare,
ns nu a influenat considerabil calitatea produselor finite analizate n cadrul
punctelor de comercializare. n special brnzeturile sunt comercializate, chiar i n
puncte de desfacere a unor super-marketuri, n condiii necorespunztoare: sunt

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expuse tipuri diferite de brnzeturi (cu sau fr mucegai n past) fr a respecta


condiii minime de prevenire a inter-contaminrii, sunt prezentate produse fr a
fi preambalate, multe produse sunt expuse n vitrine deschise, felierea produselor
se execut fr respectarea unor cerine igienice minimale.
Este important ca agenii economici care au domeniile de interes n
industria alimentar s-i nsueasc noile concepte igienice i s se concentreze
asupra ntregului lan: materie prim produs finit comercializare. Numai o
analiz dinamic a produselor lactate va asigura comercializarea unor produse
salubre, care s-i pstreze n totalitate caracteristicile igienice.

MATERIAL I METODE

Au fost prelevate probe de brnzeturi din diferite puncte de comercializare


ale Municipiului Bucureti, n lucrarea de cercetare urmrindu-se cuprinderea
unor tipuri sortimentale diferite, produse de diferii ageni economici i
comercializate n diferite uniti de profil. Pentru acest studiu au fost alese mai
multe sortimente de brnzeturi: cauri, brnzeturi srate (telemele) i brnzeturi
cu past oprit i filat, brnzeturi fermentate.
Probele prelevate au fost introduse n recipiente sterile i au fost
transportate n condiii optime de temperatur. Prelucrarea probelor a fost
efectuat n laboratorul de microbiologie alimentar conform normelor
microbiologice STAS i metodelor standardizate ISO [10, 11].
Studiul s-a desfurat pe o perioad de 1 an (n cursul anului 2005),
analizndu-se un numr total de 954 probe (125 probe de cauri, 114 probe de
brnzeturi srate din lapte de vac, 96 probe de brnzeturi srate din lapte de
oaie, 89 probe de brnzeturi srate din lapte amestec, 128 probe de cacaval
Dalia, 157 probe de cacaval Rucr, 79 probe de mozarella, 54 probe de Tilsit i
112 probe de cacaval afumat).
Pentru determinarea numrului total de germeni mezofili i aerobi, au fost
efectuate diluii zecimale n ap peptonat, din fiecare diluie s-a repartizat cu
pipete sterile cte 1 cm3 n cte 2 plci Petri. n fiecare plac s-a turnat agar topit
i rcit la 40 45 C, s-a omogenizat i s-a incubat 24 ore la 37 C. S-a
determinat media numrului de colonii / g produs [1, 3, 11].
Pentru determinarea numrului probabil de bacterii coliforme s-au efectuat
diluii zecimale, din fiecare diluie introducndu-se cte 1 ml n cte 3 eprubete cu
mediu BBLV (bulion lactozat cu sruri biliare i verde briliant) i tub Durham. S-a
incubat 24 48 ore la 37 C. Interpretarea s-a fcut dup producia de gaz i
calcularea mediei de 3 cifre (funcie de nota primit de fiecare din cele 3 eprubete
cu mediul BBLV), media obinut fiind interpretat cu ajutorul tabelului Mac
Grady. n paralel s-a utilizat i metoda de determinare a numrului de bacterii
coliforme prin numrarea coloniilor (ISO 4832). Din diluiile zecimale se
repartizeaz cu pipete sterile cte 1 ml n cte 2 plci Petri. n fiecare plac se
toarn mediu VRBL (geloz cu lactoz, sruri biliare, cristal violet i indicator),
topit i rcit la 45 C. Plcile pregtite se incubeaz la 35 C timp de 24 ore. Dup
incubare se numr coloniile rou violacee i se calculeaz media cu formula: N

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= c / (n1 + 0,1 n2) d [n care c reprezint suma coloniilor numrate; n1 este


numrul de cutii de la prima diluie reinute pentru numrare; n2 este numrul de
cutii de la a doua diluie reinute pentru numrare; d este rata diluiei care
corespunde primei diluii folosite] [1, 3, 11].
Determinarea numrului de stafilococi patogeni s-a fcut printr-o tehnic
asemntoare celei utilizate pentru determinarea NTGMA, folosind ca mediu solid
geloza Chapman sau agar Baird Parker [3, 9].
Determinarea numrului de bacterii sulfito-reductoare s-a efectuat pe
mediul cu sulfit de sodiu i citrat de fier, topit i rcit la 45 C n care s-a
nsmnat cte 1 ml din diluiile zecimale. S-a incubat n anaerobioz la 37 C
timp de 24 48 ore, dup care au fost numrate coloniile negre [1, 3, 11].
Pentru determinarea speciilor de patogeni i condiionat patogeni au fost
utilizate metodele standardizate ISO: pentru identificarea bacteriilor din genul
Salmonella SR ISO 6597, pentru identificarea stafilococilor plasmo coagulazo
pozitivi STAS ISO 6888, pentru identificarea bacteriilor din genul Proteus
SR 2356/1, pentru identificarea bacteriei Escherichia coli STAS ISO 4832,
pentru determinarea numrului probabil de E. coli SR ISO 7251, pentru
identificarea speciei Bacillus cereus SR ISO 7932, pentru determinarea
numrului de mucegaiuri STAS 12965 91, pentru determinarea numrului de
drojdii STAS 12964 91, etc [1, 11].
Toate tulpinile bacteriene izolate care au fost identificate prezumtiv ca
aparinnd unor patogeni sau condiionat patogeni au fost testate biochimic
pentru diagnostic de confirmare, datele tiinifice obinute fiind astfel riguros
demonstrate.

REZULTATE I DISCUII

Pentru a avea un element de comparaie i pentru a nelege semnificaia


rezultatelor obinute, s-a utilizat pentru interpretarea rezultatelor Ordinul
Ministrului Sntii nr. 975/1998, privind normele igienico sanitare pentru
alimente. Parametrii microbiologici prevzui n acest ordin sunt redai n
tabelul 1.
Rezultatele obinute n urma analizei bacteriologice complete a fiecrei
probe recoltate i prelucrate au fost prelucrate statistic calculndu-se medii pentru
fiecare tip de produs.
Numrul total de germeni mezofili aerobi a avut valori medii cuprinse ntre
limite relativ mari: cauri - 2,4 x 103 ; telemele obinute din lapte de vac
nepasteurizat - 1,7 x 104; telemele de oaie - 1,2 x 102; telemele proaspete obinute
din lapte amestec - 2,9 x 102; cacaval Dalia 2,1 x 102; cacaval Rucr - 4,8 x
104; Mozarella - 6,7 x 103; brnz Tilsit 2,1 x 102; cacaval afumat - 2,7 x 102.
Dei pentru tipurile sortimentale de brnzeturi analizate acest parametru nu este
normat, considerm c valorile obinute relev un numr important de
contaminani.

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Tabelul 1
Condiii microbiologice pentru alimente i materii prime
(extras din Ord. M. S. 975 / 1998)

Nr. Bacterii
Denumirea alimentului E. coli Salmonella SPCP NTDM
crt coliforme
Brnzeturi proaspete din
1. 100 10 abs. 10 -
lapte pasteurizat
Brnzeturi proaspete din
2. 1.000 100 abs. 10 -
lapte nepasteurizat
Brnzeturi proaspete: ca,
3. telemea proaspt din 10 1 abs. 10 100
lapte pasteurizat
Brnzeturi proaspete: ca,
4. telemea proaspt din 100 10 abs. 100 300
lapte nepasteurizat
Brnz proaspt din zer
5. 1.000 100 abs. 10 -
(urd)
Brnzeturi maturate n
6. saramur: ca, telemea din 10 abs. abs. 10 1.000
lapte pasteurizat
Brnzeturi maturate n
7. saramur: ca, telemea din 100 10 abs. 100 2.000
lapte nepasteurizat
8. Brnzeturi fermentate 10 abs. abs. 10 2.000
Brnzeturi cu past oprit
9. 10 abs. abs. 10 1.000
i filat
10. Brnzeturi frmntate 100 10 abs. 10 200

Analiza microbiologic pentru evaluarea numrului de bacterii coliforme,


Escherichia coli i stafilococi coagulazo-pozitivi a demonstrat o serie de depiri
comparativ cu valorile maximale admise. Analiza microbiologic pentru
evaluarea numrului de bacterii sulfito-reductoare i Bacillus cereus a
demonstrat prezena acestor specii bacteriene n probele studiate; dei acestea nu
sunt normate au fost considerate necorespunztoare probele care au nregistrat
valori mai mari dect limitele de admisibilitate stabilite la nivel european.
Numrul de probe care au nregistrat depiri i procentul de probe
necorespunztoare sunt redate n tabelul 2.
Analiza statistic a rezultatelor a permis constatarea c majoritatea
produselor au nregistrat un numr de probe necorespunztoare.

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Tabelul 2
Rezultatele analizei microbiologice a probelor de brnzeturi examinate
The microbiological analysis results for cheese product samples

Parametrul bacteriologic investigat


Bacterii
Sortimentul coliforme E. coli SPCP B. cereus Bacterii SR
Nr. Nr. Nr. Nr. Nr.
% % % % %
probe probe probe probe probe
Cauri 12 9,68 4 3,22 15 12,09 4 3,23 1 0,81
Brnzeturi
srate (lapte 17 14,78 7 6,09 21 18,26 2 1,74 5 4,35
de vac)
Brnzeturi
srate (lapte 5 5,10 1 1,02 5 5,10 2 2,04 - -
de oaie)
Brnzeturi
srate (lapte 4 4,59 - - 2 2,29 1 1,15 2 2,29
amestec)
Cacaval
2 1,57 - - 1 0,79 1 0,79 - -
Dalia
Cacaval
28 17,83 11 7,01 22 14,01 4 2,55 10 6,37
Rucr
Mozarella - - - - 14 17,95 - - - -
Brnz Tilsit - - - - 2 3,57 - - - -
Cacaval
- - - - 7 6,25 - - - -
afumat
SPCP stafilococ coagulaz - pozitiv
Bacterii SR bacterii sulfito reductoare

Din totalul probelor analizate nu au fost izolate bacterii ncadrate n genul


Salmonella. n schimb au fost izolate bacterii aparinnd genului Proteus,
Clostridium, sau avnd o ncrctur n drojdii i mucegaiuri (NTF) mai mare de
1 x 104. De asemenea la unele brnzeturi a fost constatat prezena unor specii de
mucegaiuri care fie produc compui toxici (Aspergillus), fie degradeaz structura
biochimic a proteinelor i lipidelor i modific indicatorii organoleptici (Mucor
sp.). Numrul probelor pozitive i procentul determinat din totalul probelor
analizate pentru fiecare sortiment este redat n tabelul 3.
Pe baza rezultatelor obinute n urma analizelor bacteriologice efectuate se
demonstreaz faptul c o serie de produse sunt contaminate fie n timpul
transportului, fie direct la comercializare, datorit depozitrii n condiii
necorespunztoare sau datorit meninerii la temperaturi neadecvate tipului de
produs.

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Tabelul 3
Rezultatele analizei microbiologice a probelor de brnzeturi examinate
The microbiological analysis results for cheese product samples

Parametrul microbiologic investigat


Clostridium Aspergillus
Sortimentul Proteus NTF Mucor sp.
perfringens sp.
Nr. Nr. Nr. Nr. Nr.
% % % % %
probe probe probe probe probe
Cauri - - 2 1,61 2 1,61 - - - -
Brnzeturi
srate (lapte - - 4 3,48 - - - - - -
de vac)
Brnzeturi
srate (lapte - - 1 1,02 - - - - - -
de oaie)
Brnzeturi
srate (lapte - - - - 1 1,15 - - - -
amestec)
Cacaval
- - - - - - - - - -
Dalia
Cacaval
1 0,64 2 1,27 4 2,55 1 0,64 - -
Rucr
Mozarella 2 2,56 4 5,13 - - 4 5,13 7 8,97
Brnz Tilsit 1 1,79 1 1,79 4 7,15 2 3,57 - -
Cacaval
4 3,57 2 1,79 2 1,79 2 1,79 1 0,89
afumat
NTF numr total de fungi

Prezena speciilor de Aspergillus i Mucor am constatat-o la cacavalul


Rucr i cacavalul afumat meninute la temperaturi de refrigerare (0 6 C),
acest fapt determinnd umezirea stratului protector i favoriznd, practic,
dezvoltarea sporilor de mucegaiuri (constatare frecvent la cacavalurile
meninute la temperaturi sczute mai multe sptmni) [4, 5, 6, 7].

CONCLUZII

Toate tipurile sortimentale analizate au nregistrat probe necorespunztoare


din punct de vedere microbiologic. Se remarc n mod deosebit 3 sortimente:
cauri, brnzeturi srate (telemele) din lapte de vac i cacaval Rucr, care au
nregistrat numrul cel mai mare de probe necorespunztoare, la majoritatea
parametrilor microbiologici analizai.
n urma analizei microbiologice nici un produs nu a nregistrat probe
contaminate cu germeni ncadrai n genul Salmonella.

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Examenul microbiologic pentru identificarea bacteriilor ncadrate n genul


Proteus a relevat prezena acestor bacterii n 8 probe, din care 4 probe aparinnd
unui singur tip de produs (Cacaval afumat).
Specia Clostridium perfringens a fost identificat n 16 probe (1,68% din
totalul probelor examinate), numrul cel mai ridicat de probe pozitive fiind
nregistrat de sortimentele: brnzeturi srate (telemele) din lapte de vac (3,48%)
i Mozarella (5,13%).

BIBLIOGRAFIE

1. Brzoi D., 1985 Microbiologia produselor alimentare de origine animal. Ed. Ceres,
Bucureti.
2. Brzoi D., Meica S., Negu M., 1999 Toxiinfeciile alimentare. Ed. Diacon Coresi,
Bucureti.
3. Brzoi D., Apostu S., 2002 Microbiologia produselor alimentare. Ed. Risoprint,
Cluj-Napoca.
4. Bondoc I., indilar E.V., 2002 Controlul sanitar veterinar al calitii i salubritii
alimentelor, Vol I. Ed. Ion Ionescu de la Brad, Iai.
5. Popa G., Stnescu V., 1981 Controlul sanitar veterinar al produselor de origine
animal. Ed. didactic i pedagogic, Bucureti.
6. Popescu N., Meica S., 1995 Bazele controlului sanitar veterinar al produselor de
origine animal. Ed. Diacon Coresi, Bucureti.
7. Rotaru O., Mihaiu M., 2001 Igiena veterinar a produselor alimentare. Ed.
Risoprint, Cluj-Napoca.
8. Savu C., Carmen Petcu, 2002 Igiena i controlul produselor de origine animal.
Ed. SemnE, Bucureti.
9. Tudor L., 2005 Controlul sanitar veterinar i tehnologia produselor de origine
animal. Ed. Printech, Bucureti.
10. xxx SR ISO Standarde de analize microbiologice pentru produsele de origine
animal.

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OBSERVATIONS REGARDING BACTERIOLOGICAL


AND MYCOLOGICAL MICROFLORA IN VARIOUS
CATS AND DOGS INFECTIONS
Elena VOICU, Cristina REBEGEA,
Ingrid APETREI, C. CARP-CRARE,
M. CARP-CRARE, Eleonora GUGUIANU
U.S.A.M.V. Iai

The aim of this study was the isolation and identification of


bacteriological and mycological microflora from various dogs and cats
secretions and excretions in Medical and Surgical Clinics of the Faculty of
Veterinary Medicine between 10.2005 - 04.2006.
The analizes were executed in the University Center of Medical
Research Microbiology Immunology Laboratory Iai. There were
analized a number of 17 samples of pathological matherial from 13 dogs
and 4 cats After the microbiological examination there were isolated and
identificated 27 bacteria stems and 6 mycological stems.
From the 27 bacteria stems, 9 (33,33%) of them were Escherichia
coli, 5 (18,51%) Staphylococcus spp., 5 (18,51%) Arcanobacter pyogenes,
3 (11,11%) Clostridium perfringens, 3 (11,11%) Streptococcus spp., 1
(3,70%) Pseudomonas aeruginosa, 1 (3,70%) Proteus spp. From the 6
isolated mycetes stems, 3 (50%) of them were represented Candida spp 2
(33,33%) of 2 (33,33%) Malassezia spp. and 1 (16,66%) Cryosporium spp.
For all the isolated species pathogenity tests and antibiograma were
executed. The association between bacteria and mycotic flora was observed
in 4 case, 1 case of mycotic flower only and the rest of 12 cases were with
bacteriological flora.

Key words: microbiological exam, , antibiograma, pathogenity

The microbiological flora isolated from dogs and cats with different
infections is various, bacteriological and mycotic species potentate most of the
time their pathogenity. In majority of cases pathological matherials represented
of different secretions, excretions contain an association of bacteriological and
mycotic flora. The isolation and identification of involved species is very
important establishing the diagnostic, but for the vetenarian present importance
the antibiograma results on total flora, for an efficient tratament. (1, 2, 3, 4, 6)

MATERIALS AND METHODES

Between 10.2005 04.2006 were analised 17 samples of pathological


material from 13 dogs (with ages between 7 months and 10 years) and 4 cats
(with ages between 4-9 years) of different breed: 3 samples of otical secretions, 3

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samples of pharingian exudat, 6 urine samples, 4 dermical samples and 1 sample


of vaginale secretion.
The pathological material was harvested in sterile conditions and direct
microscopic exam was made on Gram stained slides.
For microbiological flora isolation and identification the samples were
cultured on usuales and diagnostic culture mediums specific to aerobe and
anaerobe bacteria, as well as to mycetes (agarosis, bulion, Sabouraud, PDA,
mediums with potatoes extract). The tubes were incubated in thermostat at 37oC,
2448 hours for bacteria and at 22-25oC, 3-5 days for mycetes. In function of
morfoculturale characteristics there were cultered on selective and identification
mediums (Chapmann, Levine, TSI, MIU, Simmons, MacConkey).
Biochemical tests used have followed to evidence catalasyc activity,
manitol (Staphylococcus spp.), glucosis, lactosis, zaharosis fermentation, indol
production (Escherichia coli).
In species potential pathogenes it was determined the pathogenity using
haemolysis test on the sheep and rabbit erhytrocites, the test of rabbit citratated
plasma coagulation, inoculation of laboratory animals.
The sensibility to antibiotics and chimioterapics of the isolated germs was
dertermined using antibiograma, diffusimetric method (OXOID Dispenser and
antibiotics and chimioterapics microcomprimates OXOID and PHIZER: AMP
Ampicillin, AMC- Amoxycillin/Clavulanic acid, NA Nalidixic acid 30g, CN
Gentamicin 10g, E Erythromicyn 15g, K Kanamycin - 30g NOR
Norfloxacin 10g, OB Orbenin 25g, RD Rifampicin 30g, TE Tetracycline
30g, W Trimetroprim 5g), except the samples bacteriological negative
examined only mycological on native slides with lactophenol-eosine or glycerine.

RESULTES AND DISCUTIONS

The studied cases and the microbiological exam results are sinthetized in
tables 1 and 2.
Table 1
Synthesis of analised samples and the results of microbiological exam
and antibiograma in cats

Samples Cats Microbiological Antibiograma results


isolated species
Pharingiane Tomcat, 5 years - acute S - RD
E. coli
exudate laryngotraheitis R - OB, AMP, TE, E, NA
Birmanez tomcat, 9 years Proteus spp. S - RD, CN
- chronical nephritis E. coli R - NA, AMP, TE, E, K
E. coli, S - RD, K
Persane cat , 4 years
Urine Cl.perfringens R - CN, TE, W, AMC,
heamoragical cystitis
Candida spp. AMP, E
Tomcat, 5 years chronical S - W, RD, NOR
E. coli
nephritis R -TE, E, CN

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Table 2
Synthesis of analised samples and the results of microbiological exam
and antibiograma in dogs

Microbiological isolated Antibiograma


Samples Dogs
species results
Staphylococcus spp. S - CN, AMP, RD
Rottwailer, 5 years
Malassezia spp. R - K, E, TE
Otical Pseudomonas S - RD
secretions Cocker, 7 years - otitis aeruginosa R - CN, E, TE, K,
Arcanobacter pyogenes W, AMC, AMP
German shepard , 7 Cryosporium spp.,
-
years Malassezia spp.
S - RD
Cocker , 5 years -
Arcanobacter pyogenes R - CN, E,TE, K,
pharyngitis
Pharingiane AMC, AMP, W
exudate Staphylococcus spp., E. S - NOR
Ciobnesc mioritic,
coli R - TE,E, AMP,
7months
Arcanobacter pyogenes AMC, RD, W
MS - AMP, RD
Female dog , 9 years Arcanobacter pyogenes
R - OB, K, CN, NA,
cystitis Candida spp
E, TE
S - NOR
Boxer, 10 years -
Urine E. coli R - TE, AMP, AMC,
chronical nephritis
E, CN
S - AMP,
Peckinez, 3 years - Streptococcus spp.
R - TE, K, RD, E,
nephritis E. coli
AMC, W
Dog, 7 years dermatitis
Dermical Staphylococcus spp. S - OB, RD,
of bacteriological
samples Cl. perfringens R NA, AMP, TE, E
hypersensibilization
S -RD, CN
Arcanobacter pyogenes
Cocker, 1year R - E, K, TE, W,
Cl. perfringens
Raclat AMC, AMP
plantar Streptococcus S - NOR,
German shepard,
spp.Staphylococcus spp. R - W, RD, TE,
9months pyodermitis
E. coli AMC, AMP, CN, E
S -RD
Plantar Streptococcus spp.
Bull terrier, 2 years R -TE, NOR, W,
exudate Staphylococcus spp
CN, E
S - NOR
Vaginale E. coli
Female dog, 5 years R - W, RD, TE,
secretion Candida spp.
AMC, AMP, CN, E

After the microbiological exam there were isolated and identificated 27


bacteriological stems and 6 mycetes stems. From the 27 bacteria stems, 9
(33,33%) of them were Escherichia coli, 5 (18,51%) Staphylococcus spp., 5
(18,51%) Arcanobacter pyogenes, 3 (11,11%) Clostridium perfringens, 3
(11,11%) Streptococcus spp., 1 (3,70%) Pseudomonas aeruginosa, 1 (3,70%)

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Proteus spp. From the 6 isolated mycetes stems, 3 (50%) of them were
represented Candida spp 2 (33,33%) of 2 (33,33%) Malassezia spp. and 1
(16,66%) Cryosporium spp. For all the isolated species pathogenity tests and
antibiograma were executed.
Association of more bacteriological and/or mycotical species is frequent,
being observed in 5 (29,41%) of the 17 analised samples (4 of the dogs and 1 of
the cats). Staphylococcus spp. was isolated in association with Malassezia spp.,
Arcanobacter pyogenes (1 case) and Escherichia coli (2 cases) with Candida spp,
. Malassezia spp. with Cryosporium spp. (1 case).
Micoorganisms sensibility to different antibiotics and chimioterapics action
varies in function of microbiological flora association, but there are observed
differences of sensibility of defferent stems from the same identificated specie.
Results statistics sinthetized in tables 1 and 2 shows a variable sensibilitaty
of more frequent isolated species to various antibiotics and chimioterapics action:
Staphylococcus spp. (Gram positive):
- in association with Malassesia spp. is sensitive to Ampicilline action,
in the other cases being resistent (5);
- in association with Gram positive (Arcanobacter pyogenes or
Streptococcus spp.) and Gram negatives germs (Escherichia coli) is
sensitive to Norfloxacin action and resistent to Rifampicin action;
- in association with another Gram positive germ (Streptococcus spp.) is
sensitive to Rifampicin action and resistent to Norfloxacin action.
Arcanobacter pyogenes (Gram positive):
- alone or in association with other Gram positive germs (Clostridium
perfringens) is sensitive to Rifampicin and Gentamicin action towards
which Gram negativeflora is resistent.
- in association with Gram negative flora (Pseudomonas aeruginosa) is
sensitive to Rifampicin action only.
- in association with mycetes (Candida spp.) is sensitive to Ampicillin
action, towards which it it usualy resistent.
Escherichia coli (Gram negative):
- izolated in pure culture it is sensitive to Rifampicin and Norfloxacin.
- in association with Gram pozitive flora (Staphylococcus spp,
Streptococcus spp.) and mycetes (Candida spp) is resistent to
Rifampicin.
- in association with Streptococcus spp. it is sensitive to Ampicillin,
towards which it is usualy resistent.
Antibiograma results analyse reveals the fact that isolated species from all
samples are, with small exceptions, sensitives to Rifampicin and resistent to
Erythromicin, Tetracycline, Kanamycin.

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CONCLUSIONS

There were analysed 17 samples of pathological material from 13 dogs and


4 cats (3 samples of otical secretions, 3 samples of pharyngian exudat, 6 urine
samples, 4 dermical samples and 1 sample of vaginale secretion).
From the 27 bacteria stems, 9 (33,33%) of them were Escherichia coli, 5
(18,51%) Staphylococcus spp., 5 (18,51%) Arcanobacter pyogenes, 3 (11,11%)
Clostridium perfringens, 3 (11,11%) Streptococcus spp., 1 (3,70%) Pseudomonas
aeruginosa, 1 (3,70%) Proteus spp.
From the 6 isolated mycetes stems, 3 (50%) of them were represented
Candida spp 2 (33,33%) of 2 (33,33%) Malassezia spp. and 1 (16,66%)
Cryosporium spp.
Association of more bacteriological and/or mycotical species is frequent,
being observed in 5 (29,41%) of the 17 analised samples (4 from the dogs and 1
from the cats). Staphylococcus spp. was isolated in association with Malassezia
spp., Arcanobacter pyogenes (1 case) and Escherichia coli (2 cases) with Candida
spp, . Malassezia spp. with Cryosporium spp. (1 case).
Isolated bacteriological species from all the samples are sensitive, each one,
to Rifampicin (except some bacteriological associations: Staphylococcus spp.-
E.coli Arcanobacter pyogenes, Streptococcus spp. E.coli, Streptococcus
spp. E.coli - Staphylococcus spp or bacterio-mycotic: E.coli Candida spp.)
and resistent to Erythromicin, Tetracycline, Kanamycin.
Association between Staphylococcus spp. or Arcanobacter pyogenes with
Malassesia spp. or Candida spp. confers sensitivity to Ampicillin, towards which
Gram positive bacteria are resistent.

BIBLIOGRAPHY

1. Carp-Crare M., Eleonora Guguianu, Timofte Dorina, 1997 Practical works of


Veterinary Microbiology. Intern use, UAMV, Iai;
2. Guguianu Eleonora, 2002 General Bacteriology, Venus Publishing House, Iai;
3. Rucnescu H., Bica Popii Valeria, 1986 Veterinary Bacteriology, Ed. Ceres,
Bucureti
4. Rpuntean Ghe., Rpuntean S., 2005 Special Bacteriology, Ed. Academic Pres,
Cluj-Napoca.
5. ogoe I., Dobrea Mimi, 2006 Veterinary Bacteriology, Ed. Printech Bucureti.
6. Zarnea G. General Microbiology Tratate , Ed. Academiei Romne, Bucureti, vol.
I-IV, (1993-1990).

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INVESTIGAII EPIDEMIOLOGICE PRIVIND LEUCOZA


ENZOOTIC BOVIN N JUDEUL VASLUI
EPIDEMIOLOGICAL INVESTIGATIONS ON THE ENZOOTIC BOVINE LEUKOSIS
(LEUKEMIA) THROGHOUT VASLUI COUNTY

Ani DMCEANU
D.S.V.S.A. Vaslui

Investigations on monitoring the bovine enzootic leukosis (Bovine


Leukosis Virus BLV infections) throughout Vaslui county since 2000 until
2005 have been performed by serological tests of early detection of the
specific antibodies (the immuno-diffusion test in agar gel ID and the
immuno-enzymatic ELISA). During this period, a number of 427394
serological tests have been performed of which 350182 by ID and 77212 by
ID and ELISA. A number of 1643 positive samples have been diagnosed.
The incidence of BLV infections in Vaslui county recorded a continuous
dropping from 0.64% in 2000 to 0.16% in 2005. Meanwhile, a stepwise
decrease of the number of infested spots occurred, from 55 in 2000 to 14 in
2005. The spatial distribution of BLV infections throughout Vaslui county
has been monitored by drawing up cartograms. These ones highlighted the
fact that in 2000, 21 villages were free of BLV, while during the period
2001-2004 the number of free BLV villages increased to a number varying
from 37 to 41, while in 2005 the number of free BLV villages reached 62.
Thus, one can conclude that throughout Vaslui county, the degree of BLV
spreading dropped, due to the efficiency of the profilaxic measures
(draining by extraction, depopulation) enforced by the commencement of
the national program of identification, which has as goal the limitation of
the free movement of the ill animals.

Key words: BLV infections, Vaslui county, ID test, ELISA test, cartograms

Leucoza enzootic bovin (LEB) este o infecie viral produs de un virus


din familia Retroviridae, subfamilia Oncovirinae, transmisibil pe cale vertical,
de la mama la ft, ca i pe cale orizontal, de la animal la animal i de la ferm la
ferm sau de la o gospodrie la alta (1, 2, 6, 7)
Virusul leucozei bovine (VLB) produce o infecie cronic, actualmente larg
rspndit pe tot globul, dar numai 1-5% din bovinele infectate dezvolt leucemia
limfoid sau limfomul malign. Ultimele investigaii efectuate de un colectiv de
cercettori de la Universitatea din California au dovedit prezena ADN-ului VLB
n esutul mamar al femeilor bolnave de cancer mamar. Acesta constituie un un
motiv n plus pentru investigaiile epidemiologice privind LEB, n vederea evitrii
rspndirii virusului n efectivele de bovine, care constituie sursa principal de
lapte i produse lactate pentru populaie (3, 4, 5).

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In Romnia, lichidarea unitilor de cretere a animalelor organizate sub


form de CAP-uri cu preluarea necontrolat a animalelor de populaie, apoi
circulaia clandestin a animalelor, au condus la contaminarea cu VLB a
taurinelor din sectorul gospodriilor populaiei.

MATERIAL I METOD

Obiectivul prezentei lucrri este studiul incidenei i rspndirii LEB pe


teritoriul judeului Vaslui. In acest scop s-au efectuat observaii epidemiologice
prin analiza datelor din evidenele sanitar-veterinare judeene i prin efectuarea
testelor de depistare LEB incluse n Programul strategic naional de
supraveghere, profilaxie i combatere a bolilor la animale.
Dintre tehnicile serologice disponibile pentru detectarea infeciei cu VLB s-
au utilizat n cadrul Laboratorului DSVSA Vaslui, urmtoarele teste: imunodifuzia
n gel de agar (ID) i testul imuno-enzimatic (ELISA), conform metodologiei
prevzute n instruciunile oficiale (Manual OIE 2000).
Investigaiile s-au bazat pe metodele epidemiologice descriptive, lundu-se
n considerare: spaiul (ntreg teritoriul judeului Vaslui) i timpul (intervalul 2000-
2005). Rezultatele au fost prelucrate n tabele, grafice i valorificate sub form de
indicatori de frecven (prevalen, inciden) i localizare topografic
(cartograme).

REZULTATE I DISCUII

Frecvena i dinamica LEB, pe baza testelor serologice, n judeul Vaslui,


pe parcursul a 6 ani, din 2000 pn n 2005, sunt prezentate n tabelele 1, 2 i 3 i
evideniate grafic n fig. 1.
Analiza centralizat a datelor din tabelul 1 evideniaz faptul c n perioada
2000-2005 s-au efectuat 427.394 de examene serologice pentru depistarea LEB
(350.182 examene ID i 77.212 examene ID i ELISA), nregistrndu-se un total
de 1643 probe pozitive.
Urmrind evoluia n dinamic a incidenei LEB n perioada studiat se
constat c numrul animalelor serologic pozitive, raportat procentual la numrul
de animale examinate, ca i numrul focarelor nregistrate pe teritoriul judeului
Vaslui se reduc treptat de la 556 (0,64%) probe pozitive, respectiv 55 focare n
2000 la 102 (0,22%), respectiv 44 focare n anul 2005.
Intocmirea cartogramei judeului Vaslui a permis evidenierea evoluiei
focarelor de boal, precum i a gradului de infecie n focar n perioada 2000-
2005. Se remarc faptul c numrul de localiti cu grad de infecie peste 5% este
redus: 1 focar n anul 2001 la SC Cerealcom SA Roieti (6,59%) i 1 focar n
2004 la Crja (8,18%). Pe parcursul celor 5 ani investigai, numrul localitilor
indemne a crescut de la 21 n anul 2000 la 62 n anul 2005 (fig.2).

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Tabelul 1
Situaia supravegherii prin diagnostic serologic a leucozei enzootice bovine (LEB)
n perioada 2000-2005, n judeul Vaslui

Nr. probe examinate Probe pozitive


Anul
ID ID + ELISA Total Numr %
2000 64.296 22.674 86970 556 0,64
2001 99.116 - 99.116 383 0,39
2002 97.738 - 97.738 306 0,31
2003 - 54.538 54.538 171 0,31
2004 43.898 - 43.898 154 0,35
2005 45.134 - 45.134 102 0,22
Total 350.182 77.212 427.394 1.643 -

Tabelul 2
Evoluia focarelor de LEB n judeul Vaslui n perioada 2000-2005

Nr. focare Numr


Nr. CSV-uri n Numr SC Numr total Variaia
declarate curi din
care s-a n care s-a de animale gradului de
Anul infectate GP n care
diagnosticat diagn. serologic infecie n focar
LEB s-a diagn.
boala boala pozitive (%)
(GP+SC) boala
2000 55 55 529 - 556 0,09-4,65
2001 40 38 350 1 383 * 0,06-6,59
2002 36 44 304 - 306 0,03-3,39
2003 37 42 166 - 171 0,08-2,13
2004 35 44 149 - 154 0,08-8,18
2005 14 44 102 - 102 0,1-3,0
* din care 6 animale la SC Cerealcom SA Roieti, cu grad de infecie peste 5 %.

Tabelul 3
Gradul de infecie n localitile cu LEB n judeul Vaslui n perioada 2000-2005

Nr.localit. cu Nr. localit. cu


Nr. localit. Nr. localit.
Anul infecie infecie
cu infectie 1-5 % indemne
>5% <1%
2000 0 12 43 21
2001 1 6 33 36
2002 0 6 30 40
2003 0 6 31 39
2004 1 3 31 41
2005 - 4 10 62

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0,7
0,64
0,6
0,5
0,4 0,39
0,35
0,3
0,2 0,31 0,31 0,22
0,1
0
2000 2001 2002 2003 2004 2005

Fig.1. Incidena LEB n judeul Vaslui n perioada 2000-2005.

Fig. 2. Focare de LEB la sfritul anului 2005, n judeul Vaslui.


Localitile nemarcate sunt indemne de leucoz enzootic bovin.

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CONCLUZII

1. In perioada 2000-2005 n judeul Vaslui s-au efectuat 427.394 teste


serologice pentru detectarea timpurie a LEB, utilizndu-se testul de imunodifuzie
n gel de agar ID i testul imunoenzimatic ELISA. Pe ntreaga perioad
studiat s-au diagnosticat 1643 probe pozitive.
2. Incidena LEB n judeul Vaslui a nregistrat o reducere continu de la
0,64% n anul 2000 la 0,22% n anul 2005. In acelai timp, numrul de focare a
sczut treptat de la 55 n anul 2000 la 44 n anul 2005.
3. Distribuia spaial a LEB pe teritoriul judeului Vaslui s-a urmrit prin
ntocmirea de cartograme. Acestea evideniaz faptul c n anul 2000 erau libere
de LEB 21 de localiti; n perioada 2001-2004, numrul localitilor indemne a
crescut, variind ntre 37 i 41, iar n 2005 s-a nregistrat o cretere a numrului de
localiti indemne la 62.
4. In judeul Vaslui, gradul de rspndire a LEB s-a redus, datorit
eficientizrii msurilor de profilaxie (asanare prin extracie, depopulare) dublate i
de iniierea programului naional de nmatriculare (identificare), prin care se
urmrete limitarea circulaiei libere a animalelor bolnave.

BIBLIOGRAFIE

1. Arp, L.H., 1998 Leukocytic Disorders, in Merck Veterinary Manual, Eight Edition,
Merck-Co., Inc. Whitehouse Station, N.J., USA.
2. Barna, I., Gh. Prvu, C. Iugan, 1995 Leucozele i bolile de imunosupresie SIDA-
like la animale, Ed. Ceres, Bucureti.
3. Collins, D.F., H.L. Tarpley, K.S. Latimer, 2005 - A Review of Enzootic Bovine
Lymphosarcoma, College of Veterinary Medicine, The University of Georgia,
Athens.
4. Jain, N.C., 1986 - Schalm's Veterinary Hematology. Lea-Febiger, Philadelphia.
5. Mayr B., Resch S., Hepperle S., Brem G., Reifinger M., Schaffner G., 2000 -
Comparative studies in the promoter and exon 1 regions of tumour suppressor
p53 in several mammalian species: absence of mutations in a panel of
spontaneous domestic animal tumours, J Vet Med A Physiol Pathol Clin Med.,
47(10), 593-597.
6. Grimshaw, W.T., A. Wiseman, L. Petrie, I.E. Selman, 1979 - Bovine leucosis
(lymphosarcoma): a clinical study of 60 pathologically confirmed cases, The
Veterinary Record, vol 105 (12), 267-272.
7. Perianu T., S. Nicolae, M. Carp-Crare, Elena Velescu, N. Alexandru, Th.
Sptaru, Oana Tnase, 2005 Boli infecioase ale animalelor. Viroze, Vol.II,
Ed. Universitas., Iai

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INVESTIGAII PRIVIND STAREA DE PURTTOR I


ELIMINATOR DE SALMONELE MOBILE
Doina Anca HRU
U.S.A.M.V. Iai

Salmonella typhimurium is carried out in chickens intestine being


considered a transit infection because isnt located in tissues. Some
salmonellas have been isolated by different researchers from ovaries and
oviducts and in a single case from a young chicken.
Twenty healthy reproduction birds, serologically negative, were
inoculated with Salmonella enteritidis and Salmonella typhimurium.
The incubated eggs (sixty) from analyzed birds delivered thirty-two
chickens whose feathers, lachrymal secretion, brain, lungs, liver, spleen,
intestinal content and vitelus were tested bacteriological using Kaufman-
Muller and Wilson Blair growth mediums.

Cuvinte cheie: Salmonella spp., stare de purttor, paratifoz.

Din literatura de specialitate reiese c majoritatea cercetrilor sunt de acord


c psrile infectate cu salmonele mobile sunt excretoare de germeni. Totui sunt
nc discuii asupra stadiului de purttor n paratifoza ginilor i curcilor.
S-a demonstrat posibilitatea ca salmonelele s se poat transmite i vertical.
Se recomand a nu se renuna n programul de supraveghere, la controlul
serologic n efectivele infectate,cu toate c o prob serologic negativ nu denot
ntotdeauna indemnitate. Examenul serologic poate da unele indicii asupra
prezenei n efectiv al unui numr de psri infectate.

MATERIAL I METOD

Pentru a evidenia rspunsul n anticorpi aglutinani dup infecia


experimental cu salmonele i starea de purttor i eliminator, un numr de 20
gini de reproducie, serologic negative pentru tifopuloroz, n vrst de 28
sptmni, au fost inoculate cu Salmonella enteritidis i Salmonela typhimurium.
Psrile inoculate au fost controlate serologic prin reacia de hemaglutinare
cu antigen pulorum, nainte de inoculare i dup infectarea lor din 7 n 7 zile n
prima lun i din 14 n 14 zile n a doua lun de observaie.
Fiecare examen serologic a fost nsoit de efectuarea controlului
bacteriologic al excreiilor uro-digestive folosind aceleai medii uzuale, de
mbogire i selective.

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REZULTATE I DISCUII

Studiile s-au efectuat n perioada 2003 2004, n dou uniti de creterea


psrilor pentru ou de reproducie.
Din cele 20 gini, 10 au fost inoculate per os i 10 subcutanat (cte 5 pentru
fiecare specie de salmonela). Doza infectant a fost de 200 000 germeni dintr-o
cultur de 24 ore de Salmonella enteritidis i Salmonella typhimurium (tabel 1).
Cele 20 de gini inoculate cu cele dou specii de salmonela pe dou ci
(per os i subcutanat) au fost meninute n acelai adpost, iar oule produse de
ele n decurs de 60 de zile de observaie au fost recoltate n cuibare capcan,
individualizate i supuse examenului bacteriologic.
Un numr de 18 ou, produse n primele 2 zile dup inocularea tulpinilor de
salmonela au fost examinate bacteriologic prin prezena salmonelelor pe coaj i
n coninut. Alte 60 de ou, produse dup a treia zi de la infecie au fost cercetate
numai pentru prezena salmonelelor pe coaj i apoi puse la incubat. Oule clare
i embrionii mori la cele dou miraje sau la ecloziune, precum i cei rmai n
coaj au fost cercetate bacteriologic prin nsmnri directe n mediile obinuite.
Din oule puse la incubat (60 ou) au eclozionat un numr de 32 pui care
au fost sacrificai la scoaterea lor din incubator i examinai bacteriologic pentru
prezena salmonelelor pe puf, secreiile lacrimale, creier, pulmon, ficat, splin,
coninut intestinal, vitelus .
nsmnrile s-au fcut pe medii simple, cu excepia celor efectuate de pe
coaja oulor, pufului i coninutului intestinal, pentru care s-au folosit i medii de
mbogire (Kauffman-Muller) i selective (Wilson Blair).
Dup 90 zile de la infectarea lor ginile au fost sacrificate, examinate
anatomopatologic i bacteriologic.
La examenul serologic efectuat prin reacia de hemaglutinare rapid pe
lam i seroaglutinare lent n tuburi, la cele 20 de gini inoculate cu cele dou
salmonele mobile pe cale subcutanat i per os nu s-a obinut nici un rspuns
imun, prin apariia de aglutinine serice la titruri care s poat fi socotite pozitive.
O singur pasre (nr. 7) inoculat per os cu Salmonella enteritidis arat din a 14-a
si pana in a 56-a zi dup inoculare un titru serologic sczut (1/100) n reaia de
seroaglutinare lent, antigen pulorum.
n dou cazuri negative serologic prin reacia de seroaglutinare lent, s-
au nregistrat reacii slab pozitive n reacia de hemaglutinare rapid pe lam; ntr-
un caz acestea apar la 7 zile dup infecie (pasrea nr. 2), iar n altul se menin de
la 7 la 56 de zile dup infecie (pasrea nr. 9).
Stadiul de excretor de salmonele a putut fi pus n eviden la dou psri
(nr. 12 i nr. 14) inoculate pe cale subcutanat cu Salmonella typhymurium n a
14-a i a 21-a zi dup inoculare. Salmonella enteritidis folosit n infecia celor 10
gini (5 pe cale oral i 5 pe cale subcutanat), nu a putut fi izolat din secreiile

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uro-genitale, ns a fost prezent n dou cazuri pe coaja oulor produse de


psrile cu nr. 6 i nr. 17, n a 7-a zi dup infecie. Coabitarea psrilor din cele
dou grupe a dus i la realizarea infeciei cu Salmonella typhymurium a ginii
nr.5, infectat pe cale subcutanat cu Salmonella enteritidis i care a murit n a
76-a zi dup infecie din cauza unei forme septicemice de paratifoz.
La sacrificarea celor 19 gini din experien, dup 90 de zile de la
inoculare, nu s-a nregistrat nici o modificare anatomopatologic specific pentru
paratifoz, iar examenul bacteriologic al organelor parenchimatoase, ovarul i
coninutul intestinal nu au pus n eviden n nici un caz salmonele.
Examenul bacteriologic al coninutului celor 18 ou produse de 17 gini din
lotul experimental n primele dou zile dup inoculare nu a permis izolarea de
salmonele din albu i glbenu.
La examenul bacteriologic al celor 60 de ou produse de lotul experimental
din a 3-a la a 60 zi dup infecie i folosite la incubaie nu a dus la izolarea de
salmonele din coninutul oulor i nici din embrionii mori la cele dou miraje i
nici dup ecloziune.
La examenul bacteriologic al celor 32 pui eclozionai din cele 60 de ou
puse la incubat, sacrificai la scoaterea din incubator, s-a obinut: izolarea tulpinii
de Salmonella typhymurium folosit la infectarea psrilor din lotul experimental,
de la puful unui singur pui eclozionat dintr-un ou produs la 10 zile de infecie;
restul puilor eclozionai i examinai au dat n totalitate rezultate negative n ceea
ce privete prezena salmonelelor. nsmnrile efectuate de pe puful lor, din
secreiile lacrimale, organele interne parenchimatoase (ficat, splin), creier,
vitelus i din coninutul intestinal au fost sterile i negative pentru prezena
salmonelelor.
Stadiul de excretor de germeni a putut fi demonstrat la 14 i 21-a zi dup
infecia pe cale subcutanat a psrilor cu Salmonella enteritidis i o alt pasre
infectat cu Salmonella typhymurium arat c i psrile inoculate cu Salmonella
enteritidis sunt eliminatoare de germeni.
Coabitarea celor dou grupe de psri inoculate a pus n eviden caracterul
de infecie de tranzit n cazul Salmonellei typhymurium i mai ales n cazul
Salmonellei enteritidis.

CONCLUZII

Din investigaiile efectuate privind starea de purttor i eliminator de


salmonele mobile, reies urmtoarele concluzii:
1. Ginile infectate experimental au produs ou, din coninutul crora nu
s-a putut izola salmonela mobil folosit la infecie, n schimb aceste bacterii s-au
izolat de pe coaja oulor produse de aceste psri n a 10 zi de la infectarea lor, iar
din excreiile uro-genitale la 35 zile de la infecie.

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2. Coaja oulor se contamineaz mecanic cu salmonele mobile(paratifice)


dup perioada de formare.
3. Sursa principal de infecie n paratifoza aviar o constituie excretorii de
germeni, iar calea principal de transmitere este reprezentat de coaja oului
infectat n timpul trecerii lui prin cloac.
4. Msur principal de prevenire o constituie modul de pstrare a oulor
de gin destinate incubaiei.

BIBLIOGRAFIE SELECTIV

1. Brzoi D., Meica S., Negu M., 1999 - Toxiinfeciile alimentare. Ed. Diacon Coresi,
Bucureti.
2. Carp-Crare M., Guguianu Eleonora, Timofte Dorina, 1997 Lucrri practice de
Microbiologie veterinar Uz intern, Universitatea Agronomic i de Medicin
Veterinar Ion Ionescu de la Brad Iai.
3. Christensen J.P., Olson J.F., Hansen H.C., Brisgaord M., 1993 Characterization
of Salmonella enterica serovar gallinarum biovars gallinarum end pullorum by
plasmid prifiling biochimical analysis Avian Pathology (1992) 21 (3),461-470.
4. Davies R., Liebane E., Breslin M., 2003 - Investigation of the distribution and control
of Salmonella enterica serovar Enteritidis PT6 in layer breeding and egg
production. Avian Pathology (June 2003) 32(3),227-237.
5. Davies R.H., Breslin M., 2001 - Environmental contamination and detection of
Salmonella enterica serovar Enteritidis in laying flocks. - Veterinary Record, 149,
699-704.
6. Perianu T. (coord.), 2003 - Bolile infecioase ale animalelor domestice. Bacterioze.
Vol l, Ed. Venus, Iai.
7. Rapuntean Gh., Rapuntean S., 1999 Bacteriologie special veterinar
Tipografia Agronomia, Cluj-Napoca.

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CUPRINS

PR
II.. P EC
RE CL CII
NIIC
LIIN

1. Coofan V., Hricu Valentina, indilar E.V., Hricu A.C - Particulariti


morfologice ale sistemului tendo-fascial i ligamentar al autopodiului
toracic la cerb (Cervus elaphus) - Morphological particularities of the
tendo-fascial and ligamentar system of the thoracic autopodium in Cervus
elaphus ..................................................................................................................... 3

2. Belu C., Predoi G., Dumitrescu I., Georgescu B., Bioiu Carmen, Toader I.,
Pun D. - Aspecte privind morfologia unor articulaii ale membrului pelvin
la stru (Struthio camellus) - Aspects regarding the moprphology of some
articulations of pelvic limb in ostrich (Struthio camellus) ..................................... 11

3. Dumitrescu I., Predoi G., Cornil N., Belu C., Georgescu B., Dumitrescu
Florina, Rou Petronela - Aspecte histologice la nivelul reelelor
admirabile intracraniene la mamiferele domestice - The histological aspects
at the level of rete mirable at domestic mammals .................................................. 15

4. Georgescu B., Predoi G., Cornil N., Ciobotaru Emilia, Belu C., Enache L.,
Dumitrescu I., Toader A.I. - Studii privind morfostructura bursei cloacale
la palmipedele domestice - Studies concerning morphology of cloacal bursa
in domestic palmipedes .......................................................................................... 20

5. Predoi G., Belu C., Dumitrescu I., Georgescu B., Cornil N., Bioiu V.,
Toader I. - Cercetri privind vascularizaia venoas a glandei mamare la
oaie - Research regarding the venous vascularity of mammary gland in ewe ........ 27

6. Sptaru C., Sptaru Mihaela-Claudia, Ania D. - Anatomical particularities


of the pelvis of the Carpathian bear - Particularitile anatomice ale
pelvisului la ursul carpatin...................................................................................... 33

7. Sptaru Mihaela-Claudia - Particulariti morfo-funcionale ale membului


toracic la veveri (Sciurus vulgaris) - Morpho-functional particularities of
the thoracic member on the squirrel (Sciurus vulgaris).......................................... 38

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8. Sptaru Mihaela-Claudia, Sptaru C. - Particulariti anatomo- fiziologice ale


membrului pelvin la veveri (Sciurus vulgaris) - Morpho-functional
particularities of the pelvic member on the squirrel (Sciurus vulgaris).................. 43

9. Stan F., Gudea A., Damian A., Papuc I. - Particulariti ale colateralelor i
terminalelor arterelor iliace externe la ecvine - Particulariries of some
collaterals and terminal arteries of external iliac in horse ...................................... 48

10. Marcus I.B., Iancu S., Sevastre B. - Cercetri privind nivelul plasmatic al
unor componente biochimice enzimatice la obolani Wistar grefai ip. cu
ascita tumoral Walker, corelat cu consumul de ap srcit n deuteriu -
Researchs concerning the plasma level of some hepatic enzymes in Wistar
rats grafted ip. with Walker ascite tumor, correlated with deuterium
depleted water intake.............................................................................................. 53

11. Iancu S., Marcus I.B. - Investigaii biochimice privind valorile


transaminazelor serice i ale colesterolemiei corelate cu senescena la cini
clinic sntoi - Biochemical investigations regarding the values of seric
hepatic enzymes and cholesterol correlated with aging in clinically healthy
dogs ........................................................................................................................ 60

12. Priscaru Cornelia, Burlacu Anca-Irina - Evaluarea efectului


chemopreventiv al unor plante aparinnd familiei Lamiaceae Partea 1:
Aprecierea efectului chemopreventiv-hepatoprotector al unor fitopreparate
prin prisma alanil aminotransferazei si aspartat aminotransferazei - The
chemopreventive effect evaluation of some plants from Lamiaceae family
Part 1: The chemopreventive-hepatoprotector effect estimation of some
phytopreparations by point of view of alanil aminotransferase and aspartat
aminotransferase..................................................................................................... 67

13. Priscaru Cornelia, Rotaru Liliana - Evaluarea efectului chemopreventiv al


unor plante aparinnd familiei Lamiaceae Partea 2: Aprecierea efectului
chemopreventiv-hepatoprotector al unor fitopreparate prin prisma activitii
-glutamil transpeptidazei si colinesterazei - The chemopreventive effect
evaluation of some plants from Lamiaceae family Part 2: The
chemopreventive-hepatoprotector effect estimation of some
phytopreparations by point of view of -glutamil transpeptidaze and
cholinesterase ......................................................................................................... 72

14. Oancea Servilia, Creanga D., Rpa Alina, Pavel Geta - Analysis of the
natural and induced echinocytosis for two species of mammals ............................ 77

15. Oancea Servilia, Motrescu Iuliana, Rpa Alina, Airinei A. -


Spectrophotometric analysis of the blood plasma ................................................. 82

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Universitatea de tiine Agricole i Medicin Veterinar Iai

16. Falc C., Ciorba Gh., Petruse Cristina - The hematological profile of
Limousine heifers during the adaptation-acclimatization period - Profilul
hematologic la junincile de ras Limousine n perioada de adaptare-
aclimatizare ............................................................................................................ 86

17. Falc C., Ciorba Gh., Lupescu Daniela -The effects of calcium, phosphor
and of the lack of animal protein within broiler food upon some sanguine
and bioproductive parameters - Efectul nivelului calciului, fosforului i al
privrii de protein de origine animal din hrana puilor de carne asupra unor
parametrii biochimici sangvini i bioproductivi ..................................................... 90

18. Hagiu B.A., Burtan L.C., Mihailovici Maria Sultana, ura V., Ciobanu C.,
Ferariu D. - On the biocompatibility of poly (urethaneurea) doped with
Silver nanoparticles - Observaii privind biocompatibilitatea unui
poliuretanuree dopat cu nanoparticule de argint..................................................... 97

19. Lazr Roxana, Lazar M. - Cercetri privind leucocitemia, eozinocitemia i


bazofilecitemia la iepurele domestic - Researches regarding leucocytemy,
eosinocytemy and basophilcytemy at domestic rabbit ......................................... 102

20. Miclu V., Damian A., Oana L., Lisovschi C., Stan F. - Aprecieri asupra
semnificaiei morfofuncionale a complexelor neuroinsulare -
Appreciations about the morpho functional semnification of the neuro isle
complexes............................................................................................................. 109

21. Cotea C., Oprean O.Z., Solcan Carmen, Cotea I. - Structuri foliculare n
lobul intermediar al hipofizei de taur - The follicular structures in pars
intermediar of the bull hypophysis ....................................................................... 114

22. Cotea C., Oprean O.Z., Boiteanu P., Solcan Carmen, Cotea I. - Nucleii
hipotalamici la vac - The nuclei of the hypothalamus cow................................. 119

23. Solcan Carmen, Oprean O.Z., Cotea C., Solcan Gh. - Observaii privind
metaplazia cartilaginoas i osoas a pielii la vaci cu cetoz - Observations
concerning cartilaginous and osseous metaplasia of the skin in dairy cows
with ketosis........................................................................................................... 127

24. Popescu A., Cotea C., Solcan Carmen -Histochimia sinusului interdigital la
oaie - Histochemical study of the interdigital sinus (Biflexus gland) in sheep..... 132

25. Popescu A., Cotea C., Solcan Carmen - Morfologia sinusului interdigital
(glandei biflexe) la oaie - Morphology of the interdigital sinus (biflexus
gland) in sheep...................................................................................................... 144

26. Borcil Cristina, Paca S., Lazr M. - Observaii asupra unor cazuri de form
cutanat a bolii lui Marek la broileri - Observation on some cases of
cutaneous Mareks disease at broilers .................................................................. 154

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27. Lazr M., Vulpe V., Boz E., Miron L., Psrin B., Oprean O.Z. - Aspecte
lezionale ale invaziilor parazitare cu localizare branhial la ciprinidele de
cresctorie............................................................................................................. 159

28. Bulu Anca, Scripcaru C., Oprean O.Z. - Forensic examination in bitten
wounds caused by dogs to man - Expertiza medical legal n plgile
mucate provocate de cine la om ........................................................................ 165

29. Gal A., Baba A.I., Ctoi C., Iacob Sorina, Dombay E., Rus I. -Leziunile
hiperplazice/displazice adiacente tumorilor mamare maligne -
Hiperplastic/displastic lesions in tissue surrounding mammary malign
tumors................................................................................................................... 172

30. Savi B., amanc H., Stoji V., Iveti V. - Fatty liver in newborn and young
piglets ................................................................................................................... 184

31. Topal T.M., Balmu G., Cotea C., Paca S., Oprean O.Z. - Modificri
morfologice ale organelor genitale n pseudohermafroditismul masculin la
capra domestic (Capra hircus) ........................................................................... 188

32. Ursachi Gabriela, Coofan Otilia, Floritean Iulia - Fibroadenomul mamar


la pisic - Mammary fibroadenoma in cats........................................................... 194

33. Ursachi Gabriela, Coofan Otilia, Hueal G., Pavlovic I. - Mastopatia


fibrochistic la canide - Fibrocystic mastopaty in dogs........................................ 198

34. Coman M., Olariu-Jurca I., Stancu A., Rooga D. - Aspecte histopatologice
n tumorile spontane la cine - Histopathological aspects in spontan tumours
in dogs .................................................................................................................. 203

35. Olariu I., Coman M., Lazu Al., Lazu S., Stancu A. - Morphopathological
changes in the infectious peritonitis in cat and aspects related to the
histopathological diagnosis of this disease ........................................................... 208

36. Simeanu D., Stan Gh., Psrin B. - Efectul productiv al unui prebiotic asupra
performanelor productive la puii broiler de gin - The productive effect of
a prebiotic over the productive performances at boiler chicken ........................... 212

37. Curc D., Lionide A., Oranu Adriana, Constantinescu Ioana,


Samarineanu M., Cornil M. - Aspecte hematologice, biochimice i
imunologice consecutiv suplimentrii hranei cu Sel-Plex i Zn-Bioplex la
vacile lactante - The hematological, biochemical and immunological aspects
to lactating dairy cows feeded with organic zinc and selenium ........................... 216

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38. Geta PAVEL, Emil PURICE - Observaii privind variaiile profilului


hematologic la ovine cu fascioloz - On the variations of the haematological
profile in ovines with fascioliasis ......................................................................... 225

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39. Beschea-Chiriac S., Bild W., Nastasa Veronica, Haulica I. - Studiu


comparativ al reactivitii vasculare arteriale la mai multe specii de
mamifere: 1. Reactivitatea musculaturii netede arteriale la agenii
vasoconstrictori .................................................................................................... 231

40. Boghian V., Solcan Gh., Pavel Geta, Mustea M., Sauciuc R. - Clinical,
ultrasonographical and cytologic aspects of peritoneal effusion in feline
suspected of infectious peritonitis - Aspecte clinice, ecografice i citologice
ale lichidului de puncie la feline suspecte de peritonita infecioas .................... 237

41. Boghian V. - Post-partum ketonemia determined by beta-hydroxybutyrate on


Holstein frize dairy cows - Cetonemia postpartum prin determinarea
betahidroxibutiratului la vacile pentru lapte Holstein friz .................................. 241

42. Codreanu Iuliana, Dojan N., Codreanu M., Constantinescu Ioana -


Efectele deficitului hidric prelungit la cervidee, pe perioada de iarn -
Effects concerning the prolonged hidric defficiency in stags, in winter
period.................................................................................................................... 246

43. Codreanu Iuliana, Dojan N., Codreanu M., Constantinescu Ioana - Date
de profil metabolic (biochimic sanguin) la cervidee, n funcie de vrst -
Aspects concerning metabolic profile (seric biochemical) in stags,
depending on age.................................................................................................. 251

44. Codreanu M.D., Crivineanu V., Solcan Gh., Codreanu Iuliana, Cornil M.
- Aspecte clinice, hematologice i ecografice nregistrate n unele afeciuni
ale splinei la cine - Clinical, hematological and ultrasonographic
investigations in splenic diseases in dog .............................................................. 255

45. Codreanu M.D. - Rezultatele investigaiilor clinice i ultrasonografice n


diagnosticul revrsatelor cavitare la cine i pisic - Clinical and
ultrasonographic investigations in cavitary efusions in dogs and cats.................. 262

46. Goran G.V., Codreanu M.D., Crivineanu V. - Modificri hematologice,


bichimice sanguine i histopatologice induse de sulfatul de cupru la oaie -
Hematological, serum biochemical and hystopathologycal changes induced
by copper sulphate in sheep.................................................................................. 268

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47. Goran G.V., Codreanu M.D., Crivineanu V., Militaru Manuella,


Codreanu Iuliana, CIOBOTARU Emilia -Modificri histopatologice i
electronomicroscopice induse de nitratul de sodiu n experiment acut i
cronic la oaie - Hystopatologycal and electronomicroscopic changes in
acute and chronic experiment induced by sodium nitrate in sheep....................... 272

48. Papuc I., Lctu R. - Studiu comparativ privind artrografia cu substane de


contrast ionice i nonionice (Odiston i Ultravist 300) - Comparative studies
in artrography with ionic and nonionic contrast substances (Odiston and
Ultravist 300)........................................................................................................ 275

49. Hricu Luminia Diana - Observaii privind incidena sindromului urologic


felin - Observes regarding feline urologic sindrome incidence ............................ 280

50. Hricu Luminia Diana, Rileanu Gabriela - Aspecte de diagnostic n


vederea stabilirii tipului de urolitiaz la o felin domestic (studiu de caz) -
Diagnostic aspects regarding establish type of urolithiasis to a domestic
feline (case study)................................................................................................. 284

51. Sauciuc R.M., Hagiu N., Solcan Gh., Musteata M., Boghian V. - Aspects of
enzymatic diagnosis of hepato-biliary diseases in dog - Aspecte ale
diagnosticului enzimatic n afeciunile hepato-biliare la cine............................. 288

52. Rdoi I., apcaliu Agripina, Negoi Carmen, Pavel Crenguta, Condur D.
- Studiul efectelor apiterapiei cu produsul Hepro n afeciuni hepatice la
cine - A study on Hepro product api-therapeutical effects in dog liver
diseases................................................................................................................. 294

53. Costinar Luminia, Pascu Corina Mariana - Vrsta i greutatea corporal -


factori de risc pentru boala parodontal la cine - Age and body weight as
risk factors for periodontal disease dog ................................................................ 300

54. Costinar Luminia - Influenele dietei asupra bolii parodontale de la cine -


Dietary influences on periodontal disease in dog ................................................. 306

55. Miron L., Serdaru Maria, Acatrinei D. - Preliminary data regarding selenium
deficiency fodder plant from Moldova county - Date preliminare privind
deficitul de seleniu din plantele din zona Moldovei ............................................. 310

56. Miron L., Serdaru Maria, Acatrinei D. - Preliminary data regarding selenium
serical deficiency to the cattle in Moldavia - Date preliminare privind
deficitul seric de seleniu la bovinele din Moldova .............................................. 315

57. Baciu H., CozmaV., Truta Maria, Cernea M. - Testarea efectului


antieimerian al unor extracte de plante n eimerioza experimental a puilor
de prepeli - The antieimerian effects of plant extracts in quail
experimental eimeriosis........................................................................................ 319

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58. Chiimia Lidia - Relations among weight and number of eggs laid by
Dermacentor marginatus andhaemaphysalis punctata and some environment
factors ................................................................................................................... 325

59. Cristina R.T., Oprescu I., Chiimia Lidia, Dumitrescu Eugenia - In vitro
activity of a pyrethrines conditioning against Ixodidae and Argasidae ticks........ 332

60. Gudea A., Stan F., Socaciu A., Damian A. - Rezultate al investigaiilor
arheofaunale ntr-o cldire din oraului roman Napoca i o aezare rural
din localitatea Suceagu (jud.Cluj) - Results of archaeofaunal investigations
in a roman civilian building in the roman town Napoca and a rural
settlement in Suceagu village (Cluj county)......................................................... 339

61. Miron L., Miron Manuela - Food quality impact upon the relation between
length and weight of young noble crayfish - Calitatea hranei si impactul
acesteia asupra relatiei lungime-greutate la juvenilii racului de rau..................... 344

62. Miron Manuela, Miron L. - Analiza hranei utilizata de racul de rau Astacus
astacus L. din lacul Bicaz - Analysis of food utilised by the crayfish Astacus
astacus L. From Bicaz lake................................................................................... 349

62. Coofan Otilia, Iacob Olimpia, Pop I., Rileanu Gabriela - Agresiune
parazitar multipl asupra esutului hepatic la capra neagr (Rupicapra
rupicapra) - Multiple Parasitical Aggression on the Hepatic Tissueof the
Black Goat (Rupicapra rupicapra)........................................................................ 354

64. Ilie M.S., Cosoroab I., Drbu Gh., Oprescu I., Morariu S., Radbea
Narcisa, Ilie Alina, Morariu Florica - The effect of Ascaridia galli
infestation on chickens ......................................................................................... 360

65. Ioni Mariana, Mitrea I.L., Buzatu M.C. - Dinamica sezonier a


populaiilor de Ixodide din diferite zone geografice ale Romniei - Seasonal
dynamics of Ixodidae populations in different geographic area from
Romania ............................................................................................................... 365

66. Magda C., Motogna Daniela, Cozma V. - Testarea eficacitii moxidectinei


n helmintoze intestinale la galinacee - The efficacy of moxidectine in
intestinal helmintosis of Gallinaceae................................................................... 375

67. Mitrea I.L., Enchescu Violeta, Ioni Mariana, Buzatu M.C. - Isosporoza
carnivorelor domestice: aspecte terapeutice i clinic-evolutive -
Therapeutical and clincal aspects in isosporosis of domestic carnivorous ........... 381

68. Morariu S., Cosoroab I., Drbu Gh., Oprescu I., Iacobiciu I., Morariu
Florica, Radbea Narcisa - Incidena hidatidozei n sudul i vestul
Romniei n perioada 1999 2004 - The incidence of cystic echinococcosis
in western and southern Romania between 1999 and 2004.................................. 388

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69. Pavlovi I., Kuli Z., Stanojevi S. - The nematode of red foxes (Vulpes
vulpes L.) hunted in Belgrade area - Infestaia cu nematode la vulpile
(Vulpes vulpes L.) vnate n zona Belgradului .................................................... 396

70. Radbea Narcisa - Evolution of canine demodicosis in dogs without acaricid


treatment............................................................................................................... 400

71. Radbea Narcisa, Mederle O., Drbu Gh., Oprescu I., Morariu S., Ilie M.,
Izvernariu D. - Imunohistochemical results in canine piodemodicosis .............. 404

72. uteu E. - De la malarioterapie la parazitica (1927-2005): progrese n


parazitoterapie i combatere biologic - From the malario-therapy to
parazitica (1927-2005): progress in the parasito-therapy and the biological
control................................................................................................................... 407

73. Cristea Gh., Cura P., Nastasa V. - Treatment of anaerobic infections -


Tratamentul infeciilor anaerobe .......................................................................... 414

74. Munteanu N., Cura P., Cristea Gh., Nastasa V. - Classic use of local
anaesthetics in a balanced anaesthesia protocol - Utilizarea anestezicelor
locale clasice n protocolul de anestezie echilibrat ............................................. 421

75. Nastasa V., Cura P., Munteanu N., Cura Fl., Grecu Mariana -
Pharmacoclinicals considerations on the evolution of cardiovasculares
variables and acid-base balance under the combination Acepromazine-
Buprenorphine in dog - Consideraii farmacoclinice asupra evoluiei
variabilelor cardiovasculare i a echilibrului acido-bazic sub combinaia
Acepromazin Buprenorfin la cine ................................................................ 425

76. Nstas V., Nechifor Maria-Magdalena, Grecu Mariana - Evaluation of


Etomidate and Alfentanyl in dog anaesthesia - Evaluarea anesteziei cu
Etomidat i Alfentanil la cine ............................................................................. 428

77. Purice Emil, Cura P., Pavel Geta - Observaii privind efectul unor
medicamente antiparazitare asupra parametrilor hematologici la ovine cu
Fascioloz - On the effect of some antiparasitic drugs on the haematological
parameters in ovines with Fascioliasis ................................................................. 431

78. Purice Emil, Cura P., Pavel Geta, Boghian V. - Observaii privind efectul
unor substane antiparazitare asupra enzimelor serice de origine hepatic la
ovine cu Fascioloz - On the effect of some antiparasitic substances on the
serum enzymes of hepatical origin in ovines with Fascioliasis ............................ 437

79. Ciobanu S., Burtan I., Fntnaru M., Burtan L.C., Topal Roxana -
Castration of dog by scrotal septum ectomy......................................................... 443

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80. Burtan L.C., Burtan I., Fntnaru M., Ciobanu S., Topal Roxana -
Posibiliti de prevenire n neoplazia mamar la canine - Prevention
posibilities in mammary neoplasia at canines ...................................................... 446

81. Cazacu P., Mare M. - Reducerea chirurgical n hernia perineal a vezicii


urinare la un cine pekinez: raport de caz - Surgical reduction in the
perineal hernia of urinary bladder in a pekingese dog: case raport ...................... 451

82. Beteg FL., Mates N., Muste A., Oana L., Timen A - Tratamentul chirurgical
al cataractei la cine prin extracia extracapsular a cristalinului opacifiat -
Ical removal of cataracts in dogs by extracapsular extraction .............................. 461

83. Igna C., Schuszler Larisa, Sabu M. - Osinteza colului femural la caine cu
tije de acid poliglicolic - Femoral neck osteosynthesis of the dog with
polyglicolic acid rod ............................................................................................ 467

84. Muste A., Mate N., Oana L., Timen A., Beteg Fl., Ober C. - Incidena i
conduita terapeutic n parodontopatii la cine - Incidence and terapeutical
guidance parodontophaty in dogs ......................................................................... 472

85. Oana L., Timen A., Muste A., Mate N., Beteg Fl. - Poziionarea canalului
stenon cu deschiderea acestuia n sacul conjunctival - Stenon channel
transposition in the conjunctival region................................................................ 477

86. Oana L., Timen A., Muste A., Mate N., Beteg Fl. - Tehnica prostatectomiei
totale la cine - Total prostatectomy in dog.......................................................... 481

87. Timen A., Muste A., Oana L., Covaciu Monica - Utilizarea unor materiale
sintetice pentru inlocuirea ligamentelor incrucisate la caine - Synthetic
materials used for dog cruciate ligaments replacement ....................................... 485

88. Topal Roxana, Burtan I., Fntnaru M., Ciobanu S., Burtan L.C., Topal
M. - Emiologia otitei externe la carnivore - Epidemiology of otitis externa
at carnivores ......................................................................................................... 490

89. Badea M. - Observaii privind incubaia olor de stru ............................................. 495

90. Cantemir M. - Observaii privind dinamica reteniilor nvelitorilor fetale i


influena acestora asupra unor parametri de reproducie - Observations
concerning the dynamic of fetal membranes retentions and their influence
on some reproduction parameters......................................................................... 498

91. Ciornei t.G., Runceanu L., Agape G. - Cercetri privind aprecierea valorilor
rezistenei spermatozoizilor de vier - Searches about boar spermatozoa
values of resistance............................................................................................... 502

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92. Covaciu Ana- Monica, Papuc I. - Metode de determinare a momentului optim


pentru mont sau nsmnare la cea - Optimal breeding time and artificial
insemination moment in bitches, estabilished by different methods .................... 509

93. Dorobanu I., Galan C. - Observaii privind influena salpingitelor asupra


unor parametri de reproducie la vaci - Observations regarding salpingitis
influence about certain reproduction parameters of cows .................................... 514

94. Drugociu D., Runceanu L., Roca P. - The diagnostic importance of ovarian
and adenohypohysis dosage in dairy cows ovarian functional disorders.............. 518

95. Galan C., Dorobanu I. - Observaii asupra frecvenei endometritelor i


consecinele acestora n patologia reproduciei la vac - Endometritis
frequence observations and their results on reproduction pathology of cows ...... 521

96. Hromei N., Mariana Sofronie, Pntea M., Elena Ruginosu, Bdeli C.,
Anca Lupu - Studiu privind incidena mamitelor clinice la vacile cu lapte
de ras BNR - Study regarding the incidence of clinical mastitis at dairy
cows BNR breed................................................................................................... 525

97. Hromei N., Elena Ruginosu, Mariana Sofronie, M. Pntea, C. Bdeli,


Anca Lupu - Studii privind incidena mamitelor subclinice la vaci n
funcie de unii factori de variaie - The stadies regarding the incidence of
subclinical mastitis at dairy cows in function of some variation factors ............ 532

98. HUTU I. - Structuri uoare pentru ntreinerea scroafelor gestante: recenzie -


Hoop structures design for gestating sows: review .............................................. 541

99. Mircu C., Cernescu H., Otav G., Ilinca Frunz, Timea Czapp, Moisii M.,
Ardelean A. - Corelaiile evideniate ntre nivelul produciei de lapte,
performanele reproductive i nivelurile serice ale unor hormoni n
puerperiumul vacilor de lapte din rasa Blat cu Negru Romneasc -
Correlations between milk yield, reproduction and hormonal status during
post partum period in Romanian Black Spotted Dairy Cows ............................... 547

100. Roca P., Runceanu L., Drugociu D., Pachianu V. - nsmnarea


artificial intrauterin la scroaf - Intrauterine insemination of sows .................. 558

101. Runceanu L., Drugociu D., Roca P. - Cercetari asupra unor constante
sanguine in endometritele scroafelor - Researches on some sanguine
constants in sows endomethritis ........................................................................... 561

102. Vizman M.I. - Terapia reteniei placentare cu PGF2 - The therapy of placenta
retention with PGF2 ............................................................................................. 564

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103. Asociaia oamenilor de afaceri Iai 2003 - Armonizarea cu legislaia


european soluia competitivitaii pe piaa european........................................ 569

104. Brdan Gh. - Oportunitatea implementrii managementului siguranei


alimentelor n ntreprinderi ieene - Appropriateness of food safety
management implementation in Iasi enterprises................................................... 571

105. Banu Teofilia, Luchian O. - Agricultur durabil i securitate alimentar n


uniunea european. Programe de aciune adoptate de Romnia n domeniul
agriculturii durabile i a siguranei alimentelor. ................................................... 577

106. Dalea Ioana, Zegrean G. - Consideraii asupra calitii igienice a unor probe
de lapte destinate consumului public - Considerations regarding the
hygienic quality of some milk samples from public consumption ....................... 578

107. Floritean V., Elena Ian, Srbu C., Carp Crare M. - Influena
temperaturii asupra creterii tulpinilor de Bacillus cereus izolate din
alimente - Temperature influence on growth characteristics of food isolates
of Bacillus cereus ................................................................................................. 581

108. Creu Carmen, Floritean V., Carp-Crare M. - Identificarea principalelor


surse de contaminare cu microorganisme a carcaselor de pasre n abator -
Identification of the major sources of microbial contamination of chiken
carcasses in abattoirs ............................................................................................ 588

109. Brdan Gh., Psrin B. - Aplicarea sistemului HACCP ntr-un abator de


porci din regiunea de nord-est a Romaniei - Implementation of the HACCP
system in the pork slaughtering house in Nord-East region of Romania.............. 594

110. Ilie L., Savu C., Carmen PETCU - Evaluarea i controlul riscurilor asociate
procesrii crnii pentru implementarea sistemului HACCP - Control and
evaluation of risks associated with meat processing in the context of
HACCP system implementation........................................................................... 599

111. Neculi Elena Narcisa, Floritean V., Elena Ian, Carp Crare M. -
Observaii privind microflora crnii de pasre proaspete i refrigerate -
Observation concerning fresh and refrigerated poultry microflora ...................... 605

112. Neculi Elena Narcisa, Floritean V., Carp Crare M. - Influena


condiiilor de cretere i a transportului asupra calitii microbiologice ale
crnii de pasre - Influence of welfare and transportation conditions on
microbiological quality of poultry meat ............................................................... 610

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

113. Petcu Carmen, Savu C., Ilie L., Elena Mitrnescu, Camelia Papuc -
Implementarea procedurilor de igienizare n vederea certificarii sistemului
de management al siguranei alimentelor n uniti de prelucrare a petelui -
Adopting sanitation procedures in order to certify food safety management
system in fish processing units ............................................................................ 616

114. Sabu Diana, Rotaru O. - Evoluia i semnificaia numrului i configuraiei


celulelor din lapte la oaie i capr - The evolution and significance of the
somatic cell number and confugurance from goat and sheep milk....................... 621

115. Duminic Claudia Gabriela, Floritean V., indilar E. - Calitatea laptelui -


parte a conceptului ,,sigurana alimentelor n perspectiva integrrii
europene - The quality of raw milk - part of ,,food safety concept, prospect
for E.U. integration.............................................................................................. 626

116. Duminic Claudia Gabriela, indilar E. - Controlul starii de igien i a


calitii apei n unele uniti ieene de prelucrare a laptelui, efctuat prin
tehnici clasice i rapide - The quality hygiene control of work, equipament
or hand surfaces, plan enviroment and water, in some milkworks from iai
city by usual and fast microbiological methods ................................................... 634

117. indilar E.V., Totolin M., indilar E. - Nivelul unor nitrozamine volatile n
diverse produse din carne i pete afumat - The level of some volatile
nitrosamines in various meat products and smoked fish ...................................... 645

118. indilar E.V., Totolin M., indilar E. - Nivelul unor nitrozamine volatile n
unele sortimente de brnzeturi - The level of some volatile nitrosamines in
various cheeses ..................................................................................................... 651

119. indilar E.V., indilar E., Bondoc I. - Aprecierea nivelului de nitrai n


carnea proaspt de bovin, ovin, suin i peti - The estimation of nitrate
level in cow, ewe, pig and fish raw meat.............................................................. 655

120. indilar E.V., indilar E. - Evaluarea nivelului de nitrii n unele produse din
carne - The estimation of the nitrite level in some meat products ....................... 659

121. Mare M., Malic Luminia, Apetrei Ingrid, Cazacu P., Miron L. - Noi
molecule utilizate n terapia antifungic - New molecules designed for
antifungal therapy................................................................................................. 667

122. Coman I., Cuciureanu Rodica, Bulea Delia, Bobuac Maria, Teodor
Alina, Malic Luminia - Gradul de contaminare cu miceti i micotoxine a
unor substraturi vegetatale folosite n hrana omului i a animalelor .................... 676

123. Coman I., Alina Srbu, Luminia Malic, Maria Bobuac - The prevention
of fungi and mycotoxin contamination of oilseeds by chemical conservation
using volatiles reagents......................................................................................... 682

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Universitatea de tiine Agricole i Medicin Veterinar Iai

124. Furnaris F., Elena Mitrnescu, Tudor L., Tabac B., Togan C. - Cercetri
privind evaluarea bunstrii ginilor outoare exploatate n sistem
alternativ, ntr-o ferm din zona de sud a Romaniei - Researches
concerning the welfare assessment of the free-range laying hens raised in a
unit from southern Romania ................................................................................. 688

125. Georgescu Ileana, Enache L., Filipescu C., Georgescu G., Simionca I.U.,
Penariu Sanda - Unele aspecte privind aciunea aeroionizrii artificiale
negative asupra procesului infecios inflamator cutanat indus la animalele
de laborator - Some aspects regarding the negative airionization action on
the cutaneous infectious inflammation prosess induced at laboratory animals .... 694

126. Iacub G., Gurdi Viorica - Coninutul azotailor i azotiilor n raie i


concentraia lor n carne i ou la rae - The content of nitrate and nitrite in
ration and their concentration in duck meat and eggs .......................................... 702

127. Mitrnescu Elena, Tploag Dana, Ioni L., Simion Violeta, Furnaris
F., DINU Cristina - Observations concerning the air quality in the
influence area of a chemical unit - Observaii privind calitatea aerului n
zona de influen a unui combinat petrochimic .................................................... 706

128. Mitrnescu Elena, Butaru A., Tploag Dana, Tudor L., Petcu Carmen,
Staicu Elena - Observations concerning copper as a pollutant element in sol
and its influence on animal raised in a limitroph area of a chemical unit in
the south of Romania - Observaii privind cuprul ca element poluant din sol
i influiena acestuia asupra animalelor dintr-o zon limitrof unui combinat
chimic din sudul Romniei................................................................................... 710

129. ibru I. Lzrescu C. - Controlul eficienei deratizrii ......................................... 715

130. ibru I. - Ghid pentru operatorii n managementul dejeciilor solide i lichide


- Ghidance for operators on slurry and manure management............................... 721

131. ibru I. - Implicaiile ecomonice i sanitatr veterinare ale dezvoltrii


comenrului cu animale i produse de origine animal - The economic
implications of greater global trede in livestock and products ............................ 724

132 Ani Adriana Elena, Ani D.C. - Posibiliti de detecie rapid a


enterovirusurilor i teschovirusurilor suine - Possibilitys for rapid detection
of porcine enteroviruses and teschoviruses .......................................................... 731

133. Ani D.C., Ani Adriana Elena - Observaii privind prevalena unor viroze
respiratorii la bovine - Prolusions regarding the prevalence of some bovines
viral respiratory diseases ..................................................................................... 736

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

134. Ani D.C., Ani Adriana Elena - Infeciile latente cu BHV-1: mecanism i
importan - The latency of BHV-1: mechanism and importance ........................ 740

135. Cazacu P., Mare M., Cazacu Daniela - Epidemiology of exposures with
high risk for rabies infection and rabies postexposure prophylaxis...................... 745

136. Brdan Gh. - PCR detection of V. Parahaemolyticus with TLH gene


primers.................................................................................................................. 752

137. Brdan Gh. - Hybridization DNA-DNA speciesspecify method on nylon


filters for detection of V. Parahaemolyticus TLH gene ....................................... 756

138. Brdan Gh. - Real time PCR with sybrgreen to detect the pathogenically
strains of Vibrio parahaemolyticus ...................................................................... 760

139. Condrea M., Carp Crare M., Perianu T., Elena Velescu - Observations
on the frequency of Yersinia enterocolitica in pigs - Observatii privind
frecventa Yersiniei enterocolitica la suine............................................................ 764

140. Condrea M., Carp Crare M., Guguianu Eleonora, Carp Crare C. -
Some aspects regarding the methodology of isolating Yersinia
enterocolitica - Unele aspecte privind metodologia de izolare a Yersiniei
enterocolitica........................................................................................................ 768

141. Arsene M., Arsene Marinela, Savua Gh., Perianu T. - Aspecte clinice n
gripa aviar la lebada de var (Cygnus olor) - Clinical aspects of the avian
flu at the summer swan (Cygnus olor).................................................................. 774

142. Arsene M., Arsene Marinela, Perianu T., Savua Gh. - Aspecte clinice n
gripa aviar la gin - Clinical aspects of the avian flu at the chickens ............... 780

143. Carp-Crare C., Perianu T., Velescu Elena, Tnase Irina-Oana, Pavli C.,
Ciobanu G. - Aspecte epidemiologice i clinice n listerioza ovin ................... 785

144. Carp-Crare C. - Observations regarding the isolation of Listeria


monocytogenes in silo fodder ............................................................................... 792

145. Chiriac Adriana, Carp- Crare M., Neculi C. - Diagnostic de laborator


complex al tuberculozei la bovinele sacrificate - The complex diagnosis of
tuberculosis infection at the sacrificed cattle........................................................ 797

146. Cadar D., Spnu Marina, Miclu V., Kblkuti L., Tuboly T., Czirjk
G., Dn ., Csgola A. - Sindromul dermatitei si nefritei porcine (PDNS)
o nou entitate morbid n patologia suin din Romnia - Porcine dermatitis
and nephropathy syndrome (PDNS) a new pathological entity in
Romanian swine herds.......................................................................................... 804

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Universitatea de tiine Agricole i Medicin Veterinar Iai

147. Ctan N., Herman V., Necsulescu M., Fodor I. - Testarea clinic a unui
vaccin polivalent inactivat destinat imunoprofilaxiei unor clostridioze
bovine - The clinical trials of inactivated polyvalent vaccine used in the
specific immunoprofilaxy of some bovine clostridiosis. ...................................... 809

148. Hru Doina Anca, Perianu T., Tnase Irina-Oana, Pavli C. - Observaii
privind etiologia paratifozelor aviare ................................................................... 816

149. Ciocrlan Emilia, Perianu T, Tnase Irina-Oana, Pavli C. - Determinarea


patogenitii speciilor bacteriene implicate n patologia infecioas de
incubaie - Eggs incubation pathology and pathogenicity of the bacteria
isolated ................................................................................................................. 821

150. Ian Elena, Merticariu tefania, Floritean Iulia, Burtan Eugenia - Cteva
aspecte virusologice n gripa aviar - Some virusological aspects in bird flu ...... 824

151. Rebegea Cristina, Carp-Crare M., Carp-Crare C., Anton C., Fodor D.
- Epidemiological screening over the Mycoplasma hyopneumoniae wearers
in two swine breeding units.................................................................................. 834

152. Savuta Gh., Stoichici A., Anita D., Anita Adriana, Butunoi C., Popovici-
Bibirus Roxana - Evaluarea epidemiologica si economica a programului
de combatere a tuberculozei bovine in Romania - Epidemiological and
economical evaluation of bovine tuberculosis in Romania .................................. 840

153. Savua Gh., Matei Irina, Murat Selma, Ania D. - Epidemiologia


Paratuberculozei rumegatoarelor n Romnia - Epidemiology of ruminants
Paratuberculosis in Romania ................................................................................ 845

154. Starciuc N., Scutaru I., Zelenin S. - Modificarea structurii bursei lui
Fabricius sub aciunea tulpinilor vaccinale fierbini contra bursitei
infecioase aviare. - The actions of hot strain BG against IBD to
structural modifications of tissues of cloacal bursa. ............................................ 850

155. Neculi C., Carp Crare M., Neculi Narcisa, Chiriac Adriana - Aspecte
epidemiologice privind Bruceloza ovin n judeul Vaslui - The
epidemiological aspects about the infection with Brucella ovis in Vaslui
county ................................................................................................................... 852

156. Bradatan Gh., China B., Daube G. - Real-time PCR Assay with Taqman
probe to detect the tlh gene of V. Parahaemolyticus ............................................ 857

157. Chiriac Adriana, CarpCrare M., Neculi C. - Sensibilitatea i


specificitatea unor teste de diagnostic serologic utilizate n tuberculoza
bovin - The sensibility and the sensitivity of the serologycal diagnosis
tests used in the diagnosis of the bovine tuberculosis .......................................... 862

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Lucrri tiinifice vol. 49 seria Medicin Veterinar

158. Guguianu Eleonora, Bltescu Ana-Maria - Investigatii privind calitatea


microbiologic a laptelui comercializat de productorii particulari -
Investigations regarding the microbiological quality of the milk sold by the
privat producers.................................................................................................... 868

159. Ili ., Spalevi L.J., Miljkovi B., Pavlovi I., ugi G., Stanojevi S. -
Efficacy of same antimicrobial substance at pathogen microorganisms
isolated in poultry breeding objects ..................................................................... 872

160. Tudor L., ogoe I. - Evaluarea izolrii speciilor Vibrio non-holerice din
probe de ap prelevate din sudul i sud-estul rii - The isolation evaluation
of non-choleric Vibrio species in south and southeast Romanian waters............. 875

161. Tudor L., ogoe I., Mitrnescu Elena - Evaluarea calitii microbiologice a
unor brnzeturi comercializate pe pieele bucuretene - The evaluation of
microbiological quality for some types of cheese traded on Bucharest
markets ................................................................................................................. 880

162. Voicu Elena, Rebegea Cristina, Apetrei Ingrid, Carp-Crare C., Carp-
Crare M., Guguianu Eleonora - Observations regarding bacteriological
and mycological microflora in various cats and dogs infections.......................... 887

163. Dmceanu Ani - Investigaii epidemiologice privind Leucoza enzootic


bovin n judeul Vaslui - Epidemiological investigations on the Enzootic
bovine leukosis (Leukemia) throghout Vaslui county.......................................... 892

164. Hru Doina Anca Investigaii privind starea de purttor i elimonator de


salmonele mobile.................................................................................................. 897

917
Consilier editorial Vasile VNTU
Tehnoredactor: Rodica TRIFAN
Corector: Liviu MIRON
Gheorghe SOLCAN

Bun de tipar: 30.10.2006


Aprut: 2006, 70x100/16
Editura: Ion Ionescu de la Brad Iai
Aleea M. Sadoveanu nr. 3
Tel.: 0232-218300
E-mail: editura@univagro-iasi.ro

ISSN: 1454-7406

Tiparul executat la PIM Iai

PRINTED IN ROMANIA

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