Collection,Preservation & Despatch of Clinical Materials മലയാളം

മലയാളം

GENERAL CONSIDERATIONS FOR COLLECTION OF SPECIMENS

1. The diseases most commonly encountered in animals are of bacterial, viral,

parasitic, fungal and metabolic origin. Diagnosis based on symptoms and
laboratory examination of the relevant materials is essential for initiating treatment
at the proper time. In general the following points should be duly considered while
collecting materials for laboratory diagnosis.
2. All materials collected should be accompanied by full history of disease out break
namely species affected, duration of disease, clinical signs, morbidity and
mortality rates, disease suspected etc.
3. The collected biological specimens should be transported on ice to the nearest
laboratory as early as possible.
4. Materials collected for bacteriological examination should be kept at refrigeration
temperature (4o C) in case of delay of transportation. If a viral etiology is
suspected the material can be stored at –20 to -80o C.
5. When sero-diagnosis is required, collect paired serum samples (about 2 ml sera).
One serum sample should be collected at the onset of disease and second sera after
recovery (3-4 weeks) from disease preferably on 21st day.
6. If death is reported, the post-mortem examination should be conducted at the
earliest as putrefied materials are unfit for laboratory examination.
7. Detailed post-mortem report should be attached along with the samples collected
during postmortem.
8. The different virological transport media that can be used are 50% Phosphate
Buffered Glycerine Saline and Phosphate Buffer Saline (pH 7.2-7.4). Collect
samples in sterile containers when a transport media is not available and culars
them on ice as early as possible.
9. For Histopathology studies, tissues should be preserved in 10% formalin. The
volume of formalin used should be approximately 10 times the volume of material.
Specimen bottles with wide mouth should be used for collecting tissues.
10. The specimen bottles should be sealed well so as to avoid leakage and cular
clearly indicating the fixative/transport media used.
11. All the impression smears before culars, should be fixed in methanol for 1-5
minutes unless otherwise specified.
12. In case of outbreaks, try to collect materials from as many ailing animals (5-6 or
more) as possible at the height of body temperature /clinical signs.

The type of materials to be collected in different disease conditions are mentioned below

BACTERIAL DISEASES

HAEMORRHAGIC SEPTICAEMIA

Caused by Pasteurella multocida and characterized by high fever, localized oedema and
respiratory symptoms.
Materials to be collected

From ailing animals

1. Blood smear
2. Smears of fluid from swelling
3. Blood in sterile container

From dead animals-

1. Impression smear from heart, lungs, liver, submaxillary swellings

2. Smears from heart blood
3. Lymph nodes and spleen on ice

ANTHRAX

A zoonotic disease caused by Bacillus anthracis. Disease is characterized by high fever,

bloat, respiratory distress due to oedema of thorax and brisket region, muscular tremors,
abdominal pain and sudden death followed by bloody discharges from natural orifices.
Materials to be collected

1. Blood smears from ear vein

2. Smear of the discharges from natural orifices
3. Smear from swelling
4. Ear tip or a piece of muzzle in saline

Note: Do not open a carcass if anthrax is suspected. Such carcasses should be disposed by
deep burial with quick lime and the premises should be thoroughly cleaned with
disinfectants such as 10% formalin or 10% caustic soda.

BLACK QUARTER

An acute infectious disease caused by Clostridium chauvoei . Disease is characterized by

development of focal gangrenous and emphysematous myositis.
Materials to be collected

1. Impression smears from the affected muscle.

2. Exudate from the swelling on ice
3. Pieces of affected muscle on ice

ENTEROTOXAEMIA, LAMB DYSENTERY

An infectious disease of ruminants caused by Clostridium perfringens. Disease is

characterized by abdominal pain, hemorrhagic enteritis and sudden death. Symptoms vary
depending upon the type of toxin produced by the organism (types A, B, C, D, E, F etc.).

Materials to be collected

1. Blood
2. Smears from contents of small intestine
3. Contents of intestine

Contents of small intestine with and without chloroform separately on ice, kidney, urine.

BRUCELLOSIS

Contagious abortion caused by Brucella abortus. Materials to be collected

1. Milk
2. Blood
3. Serum sample (Paired serum sample)
4. Vaginal mucus, uterine fluid.
5. Stomach contents of foetus, aborted foetus

JOHNE’S DISEASE

A chronic, infectious, fatal gastrointestinal disease of ruminants caused by

Mycobacterium johnei. The most cardinal symptom is continuous or intermittent culars
leading to progressive emaciation and death.
Materials to be collected

1. Rectal pinch swab or smear

2. Faecal sample
3. Terminal portion of ileum with ileocaecal valve, mesenteric lymph gland in 10%
formal saline.

GLANDERS

A zoonotic disease usually seen in horses caused by Actinobacillus mallei (Pseudomonas

mallei). The disease is usually seen as a chronic condition characterized by nasal
discharge, formation of small nodules on upper respiratory tract mucosa and along the
lymphatic channels of the skin and presence of ulcers on the skin.
Materials to be collected

1. Nasal discharge on ice

2. Pus or culars from skin lesions on ice
3. Impression smears of culars
 4. Affected tissue for histopathology

TUBERCULOSIS

A chronic contagious disease of man and animals caused by different species of the Genus
Mycobacterium. The disease is characterized by a painful, dry, hacking cough, respiratory
distress, abdominal pain, diarrhea, chronic bloat, emaciation, irregular oestrus cycle,
abortion, sterility, formation of small nodules in mammary tissues, painful swellings of
the joints etc., depending upon the involvement of the different systems of the body.

Materials to be collected

1. Sputum swabs in sterile container on ice

2. Milk in sterile vials on ice
3. Faeces
4. Heat fixed impression smears from lymph glands
5. Affected tissue for histopathology in 10% Formalin
6. Lymph glands or lung lesions in sterile container in 50% Glycerol Phosphate
buffer.

LEPTOSPIROSIS

A zoonotic disease caused by the different species of the Genus Leptospira. The disease is
seen as an acute or chronic or clinically inapparent condition and is characterized by
sudden fever, muscle tremors, anorexia, haemoglobinuria, icterus and abortion.

Materials to be collected

1. Blood
2. Blood smear
3. Serum
4. Urine
5. Tissue from kidney, liver and spleen in 10% formalin
6. Milk or urine in vials by adding 1 drop of formalin per 20 ml

SALMONELLOSIS

A chronic or acute contagious disease of all species of animals, birds and human beings
caused by the various serotypes of the Genus Salmonella. Disease is characterized by
gastroenteritis, high temperature, dullness and ataxia. Usually the disease is manifested in
two forms namely enteric form and septicaemic form.

Materials to be collected
 1. Blood
2. Faeces
3. Intestinal content, heart blood and bile in separate sterile vials
4. Tissues like mesenteric lymph nodes, kidney and gall bladder for histopathology

ACTINOMYCOSIS

Actinomycosis or Lumpy jaw, a chronic infectious disease, affecting cattle primarily is

caused by Actinomces bovis. The disease also affects pigs, horses and human beings and
is characterized by osteomyelitis of the bone of the skull in cattle, enlargement of the
mammary gland in pigs and involvement of the bursa of ligamentum nuchae in horses.
Materials to be collected

1. Pus smear
2. Pus in sterile container on ice
3. Tissue from swelling for histopathology

ACTINOBACILLOSIS

Actinobacillosis or Wooden tongue is a chronic infectious disease of cattle and sheep and
is caused by Actinobacillus lignieresi. The disease is characterized by inflammation of the
tongue and soft tissues around the head and neck, protrusion of the tongue that is hard and
swollen, anorexia, progressive weight loss, enlargement of submandibular lymph nodes
and a loud snoring sound.
Materials to be collected

1. Pus smear
2. Pus in sterile container on ice
3. Affected tissue for histopathology

LISTERIOSIS

Listeriosis or Circling disease is a fatal infectious disease of man and animals caused by
Listeria monocytogenes. The disease is characterized by encephalitis, abortion or
septicaemia.
Materials to be collected

1. Blood
2. Cerebrospinal fluid
3. Brain
4. Aborted foetus or placenta
5. All internal organs in 10% formalin or on ice.
MYCOPLAMOSIS/CCPP/CBPP/CORYZA

Contagious disease of cattle, sheep, goat and poultry affecting thoracic organs especially
lungs and pleura. Disease is caused by the different species of the Genus Mycoplasma.
Materials to be collected

1. Smears from culars

2. Swabs from sinus/trachea
3. Nasal and vaginal swabs in PBS on ice
4. Piece of lung preserved in 10% formalin and on ice separately
5. Serum samples (paired serum)

CHLAMYDIA/PSITTACOSIS

Materials to be collected

1. Nasal swab
2. Lung pieces in sterile vials on ice
3. Impression smears from lungs, liver, conjunctiva and foetus
4. Serum samples (Paired serum samples)
5. Internal organs in 10% formalin

BOVINE GENITAL CAMPYLOBACTERIOSIS

Contagious venereal disease of cattle characterized by abortion, infertility with repeat

breeding. Caused by Campylobacter foetus.

Materials to be collected

1. Vaginal mucus
2. Stomach content of aborted foetus

VIRAL DISEASES

BLUE TONGUE DISEASE

From ailing animals –Blood at the height of body temperature in heparin or EDTA for
isolation. Paired sera in sterile vials on ice for serology

From dead animals –Spleen, lung and lymph nodes on ice for isolation. Spleen, lymph
nodes, intestine in 10% formol saline for HP
BOVINE VIRAL DIARRHOEA/ MUCOSAL DISEASE

From ailing animals –Nasal / cular discharge for isolation, Blood in EDTA for isolation,
Paired serum samples for serology, Semen for isolation. From dead animals –Lymph
nodes and spleen in sterile vials on ice for isolation, Intestinal swabs for isolation

CANINE DISTEMPER

From ailing animals –Blood smears / Impression smears of conjunctiva or vaginal

epithelium for viral inclusion body. From dead animals –Impression smears from liver for
viral inclusion body, Pieces of liver & spleen in sterile vials on ice for isolation, Pieces of
lung, urinary bladder, liver, trachea, stomach wall and brain in 10% formol saline for HP

FOOT AND MOUTH DISEASE

From ailing animals –Vesicular epithelium in 50% Phosphate Buffered Glycerine, Oeso-
pharyngeal fluid in 50% Phosphate Buffered Glycerine

INFECTIOUS BOVINE RHINOTRACHEITIS (IBR)

From ailing animals Paired

serum for serology, Swabs from vaginal and nasal lesions for isolation. From dead
animals –

Pieces of trachea, lungs in PBS on ice for isolation, Pieces of trachea, liver, turbinate
bone,

lungs for HP

INFECTIOUS CANINE HEPATITIS

From dead animals –Spleen and liver in sterile containers on ice for isolation. Liver, gall
bladder and kidney in 10% formol saline for HP. Impression smears from liver fixed in
methanol

PARVO VIRAL INFECTION OF CANINES

From ailing animals –Rectal swab and faeces in PBS, Paired serum samples. From dead
animals –Pieces of intestine, heart on ice for isolation. Liver, heart, intestine in 10%
formol saline for HP.

PESTE DES PETITS RUMINANTS (PPR)

From ailing animals –Eye, mouth and

rectal swabs in PBS on ice for isolation. Citrated Blood for isolation. Serum for serology.
From dead animals Spleen, lymph nodes, pieces of intestine on ice for isolation. Lungs,
liver, spleen, tonsil in 10% formalin for HP

RABIES

From dead animals –Head / Whole carcass on ice for demonstration of viral antigen, viral
inclusions and isolation of virus. Brain on ice for demonstration of viral antigen, viral
inclusions and isolation of virus. Note: It is not advisable to open the skull by persons not
protected by vaccination.

RINDERPEST

From ailing animals –Blood for isolation. Paired Serum samples for serology. From dead
animals –Spleen lymph node, alimentary tract mucosa for HP

SWINE FEVER

-From ailing animals –Heparinised blood at the height of temperature for isolation. From
dead animals Pieces of spleen, mesenteric lymph glands, intestine especially ileocaecal
region in 50% glycerol saline for isolation. Pieces of brain, lung, intestines, ileocaecal
region and kidney for HP

POULTRY DISEASES

INFECTIOUS BURSAL DISEASE (GUMBORO DISEASE)

From ailing animals –Ailing bird. Paired serum sample. From dead animals –Bursa of
fabricious in 50% Buffered Glycerine saline. Bursa of fabricious, kidney, spleen for HP

MAREK’S DISEASE

From ailing animals –Ailing bird. Paired serum sample. From dead animals –Trachea,
ovary, liver, kidney, spleen for HP

RANIKHET DISEASE

From ailing animals –Serum sample, Ailing bird. From dead animals –Fresh carcass on
ice. Liver, spleen, trachea, bronchi, lungs in 50% Buffered Glycerine saline.
Proventriculus for HP

PARASITIC DISEASES

THEILERIOSIS
-Blood smears, Biopsy smears from swollen lymph nodes from early stage of disease
fixed with Methanol
BABESIOSIS-Thin blood smears fixed in Methanol 3. ANAPLASMOSIS- Thin blood
smears fixed in Methanol

FILARIASIS

Direct blood film (wet film), Blood smear

SURRA/TRYPANOSOMIASIS

Wet film examination of blood by hanging drop,Fixed blood smears, Blood in

anticoagulant on ice 5.

SCHISTOSOMIASIS-

Dung sample (Avoid contact of water which may aid in hatching of the egg). Nasal
schistosomiasis –Nasal discharge in normal saline, Nasal granuloma in normal saline

TRICHOMONIASIS

-Vaginal or uterine discharges, Prepucial scraping/ washing

GASTRO-INTESTINAL PARASITIC DISEASES

-Faecal sample, Affected internal organs in 10% formalin

ECTOPARASITIC INFESTATIONS (RINGWORM, MANGE, MITES)

Deep skin scrapings in sterile vials

MYCOTIC / FUNGAL INFECTIONS

RHINOSPORIDIOSIS

-Nasal discharge in sterile vial, Nasal swab, Granulation tissue in 10% formol saline

EXTERNAL FUNGAL INFECTIONS

Skin scrapings in sterile vials

TOXICOSES/ POISONINGS

AFLATOXICOSIS

-100 gm of suspected feed (specially groundnut cake), Liver and spleen in 10% formol
saline, Liver and spleen on ice

POISONING CASES

For chemical analysis fresh tissues and fluids should be sent as soon as possible and on
ice. Avoid addition of preservatives to the samples. Use 95% ethanol @ 1ml per gram of
sample when necessary.

PLANT POISONINGS

Sample of suspected grass/fodder/plants, Liver on ice, Stomach contents on ice.

MISCELLANEOUS CONDITIONS

MASTITIS

Milk samples (mid-stream) before onset of treatment in sterile vials on ice.

ABORTION

Whole foetus/ all internal organs of foetus on ice, Vaginal swab in PBS, Pieces of
placenta in sterile vials on ice, Pieces of placenta in 10% formalin, Paired serum samples

ETHMOID TUMOU

R-Tumour affecting mucosa of ethmoid turbinates of cattle, Nasal discharge in sterile

vials, Deep nasal swab, Tumour tissue in 10% formol saline

INFERTILITY AND STERILITY

Semen in sterile vials, Prepucial swab on ice, Paired serum sample on ice

PYREXIA OF UNKNOWN ETIOLOGY

-Blood smears, Blood collected in EDTA, Paired serum sample on ice

DISEASES OF UNKNOWN ETIOLOGY

-Feed/ Fodder, Blood smears, Urine sample, Faecal sample, Blood samples collected in
EDTA on ice from live animals, Serum samples from live animals, Stomach contents,
spleen, lung, lymph node, liver, kidney, intestine in sterile vials from dead animals on ice,
Stomach contents, spleen, lung, lymph node, liver, kidney, intestine in 10% formol saline

USGS – National Wildlife Health Center

INSTRUCTIONS FOR COLLECTION AND SHIPMENT OF AVIAN AND
MAMMALIAN CARCASSES
Contact your USGS Field Investigation Team (FIT) member first!

Western States: Krysten Schuler kschuler@usgs.gov, 608-270-2447

Central States: LeAnn White clwhite@usgs.gov, 608-270-2491
Eastern States: Anne Ballmann aballmann@usgs.gov, 608-270-2445
Hawaii, Pacific Islands: Thierry Work thierry_work@usgs.gov, 808-792-9520

For single animal cases

Nationwide: Jennifer Bradsby jbradsby@usgs.gov, 608-270-2443

Emergency Contact Number 608-270-2400

The following instructions should be used for collecting and shipping wildlife carcasses, carcass parts, and
samples extracted from animals to the National Wildlife Health Center (NWHC) in Madison, Wisconsin, to
insure adequate and well preserved specimens. For shipping instructions to the Honolulu Field Station,
please contact Thierry Work (please see contact information above).

Freezing/thawing impedes isolation of some pathogens and damages tissues. NWHC prefers unfrozen
specimens if they can be sent within 24-36 hours of collection or death. We will provide guidance on
freezing samples on a case-by-case basis. As a general guideline: if you cannot call or ship within 24-36
hours, freeze the animal(s).

□ Contact FIT to get shipping approval and discuss shipping arrangements. Typically, ship
specimens by 1-day (overnight) service, Monday through Wednesday, to guarantee
arrival at NWHC before the weekend. If specimens are fresh and need to be shipped on
Thursday or Friday, special arrangements can be made.

□ Email/fax history and tracking number to FIT. Packages will not be opened if history
does not arrive first!

□ Use rubber, vinyl, or nitrile gloves when picking up sick or dead animals. If you do not
have gloves, insert your hand into a plastic bag.

□ More than one disease may be affecting the population simultaneously. When possible,
collect both sick and dead animals. Note behavior of sick animals before euthanizing.

□ Collect specimens that are representative of all species affected and geographic areas.

□ Collect the freshest dead specimens. Decomposed or scavenged carcasses are usually of
limited diagnostic value. If you plan to collect animals in the field, take along a cooler
containing ice to immediately chill carcasses.

□ Contact NWHC for assistance when collecting samples from animals that are too large to
ship.

□ Collect animals under the assumption that an infectious disease or toxin is involved and
other animals may be at risk. Protect yourself as some diseases and toxins are hazardous
to humans.

□ Immediately attach a leg tag to each animal with the following information in pencil or
waterproof ink:
- Date collected - Species
- Location (specific site, town, county, state) -Found dead or euthanized
- Collector (name/address/phone) -Your reference #
 □ Place each animal in a plastic bag, close, and seal the bag. Cover zipper bag closure with
strapping or duct tape after sealing zipper.
Twist non-zipper bags closed, fold over on itself, and secure with package strapping or duct tape.

□ Place 1st bag inside a 2nd bag, close and seal. More than one individually bagged animal
can be placed in the 2nd bag. This prevents cross-contamination of individual specimens
and leaking shipping containers.

□ Tag the outside of 2nd bag and number of animals and type, date collected, location, and
name of collector. Reminder order: TAG, BAG, BAG, TAG.

□ Use a hard-sided cooler in good condition for shipment. Close the drain plug of cooler
and tape over inside. Line cooler with a thick bag (1 mil thickness, 3rd layer of bags).

□ Place absorbent material in the 3rd

plastic bag to absorb any liquids that
might leak during shipping.
See appendix for examples of bags
and absorbent materials.

□ Pack the individually bagged

animal(s) that are contained within
the 2nd sealed bag into the 3rd bag
with enough FROZEN BLUE ICE
PACKS or similar coolant to keep
carcasses cold. Use enough coolant to keep samples chilled if there is a
delay in delivery.
o Blue ice (unfrozen) can be obtained at hardware, sporting goods, or
grocery stores.
o Wet ice can be used if frozen in a sealed plastic container (i.e., soda or
water bottle).
o DO NOT USE DRY ICE.
□ Seal the 3rd bag with methods described for 1st bag.
□ Place the completed specimen history and return shipping label in a ziplock bag and tape
to the inside lid of the cooler (if you want the cooler returned). NWHC CANNOT PAY
FOR SHIPPING.

□ Using packing or duct tape, tape the cooler shut around the lid and at each end using a
continuous wrap around the cooler.

□ Attach the shipping document (airbill) with the DOT information below to the outside of
each cooler in a resealable pouch:
Address:
National Wildlife Health Center
Necropsy Loading Dock
6006 Schroeder Road
Madison, WI 53711
Emergency Contact:
NWHC FIT emergency
608-270-2400
Supplementary Labels:
Keep Cold
□ Mark the cooler with the appropriate information:
 (See Pg. 3 for printable marking labels)
o Carcasses of animals that died of unknown causes:
BIOLOGICAL SUBSTANCE, CATEGORY B and UN 3373.
o Blood and tissue samples from apparently healthy animals (hunter-killed, live captured):
EXEMPT ANIMAL SPECIMENS.
o Blood and tissue samples from dead or sick animals:
BIOLOGICAL SUBSTANCE, CATEGORY B and UN 3373.
.
□ Note the tracking number in case packages are delayed.

□ These instructions cover federal shipping regulations for commercial carriers.

Appendix:
Example of bags available at large supermarkets (list not all inclusive):

Inner and second layer bags:

Hefty Big Bag – 22 gal Ziplock Freezer – 1 gallon
Hefty Freezer – 1 gal Ziplock Big Bag – 20 gallon
Hefty Jumbo – 2.5 gal Glad Freezer – 1 qt, 2 qt, 1 gal

Third layer for cooler liner:

Hefty Cinch Sak (1.1 mil) – 33 and 39 gal Glad Force Flex (1.05 mil) – 25 gal
Hefty Lawn and Leaf (1.1 mil) – 33 and 39 gal Hefty Ultra Flex (1.3 mil) – 30 gal
House brand large trash (1.1 mil) – 30 gal House Lawn - Leaf (1.2 mil) – 39 gal

Absorbent material:
Super absorbent packet or pads for water Cellulose wadding
Paper towels Cotton batting or cotton balls
Do not use packing peanuts or shredded paper.

UN3373
 BIOLOGICAL SUBSTANCES, CATEGORY
B

EXEMPT ANIMAL
SPECIMENS