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Small Ruminant Research 97 (2011) 110116

Contents lists available at ScienceDirect

Small Ruminant Research

j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / s m a l l r u m r e s

Respiratory syncytial and parainuenza type 3 viruses coexisting with

Pasteurella multocida and Mannheimia hemolytica in acute pneumonias
of neonatal alpacas

R. Rosadio a,b, , E. Cirilo a , A. Manchego a , H. Rivera
a

a
Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Avenida Circunvalacin 2800, San Borja, Lima,
Peru
b
CONOPA (Instituto de Investigacin y Desarrollo de Camlidos Sudamericanos), Los Cerezos 106, Ate Vitarte, Lima,
Peru
 Author's personal copy

a r t i c l e i n f o a b s t r a c t

Article history:
The etiopathogenesis of acute pneumonia, the second most important cause of mortality
Received 10 June 2010
among neonatal alpacas in Peru, is still poorly understood. The objective of this study was
Received in revised form 20 January 2011
Accepted 2 February 2011
to characterize gross and histopathology lesions, as well as to identify viruses
Available online 2 March 2011 [parainuenza type 3 (PI-3) and/or bovine respiratory syncytial (BRS)] by direct
inmunouorescence test, and isolate bacteria [Pasteurella multocida (Pm) and/or
Key words:
Mannheimia haemolytica (Mh)] from 24 fatal acute pneumonia cases of alpaca
Neonatal alpaca neonates 739 days old. At necropsy the gross lesions corresponded to moderate
Acute pneumonia purulent focal bronchopneumonia or severe necrotic purulent brinous (n = 13), and
PI-3 virus moderate to severe pulmonary congestion and/or pulmonary edema (n = 11).
BRSV Histopathological analysis revealed: acute severe and diffuse necrotizing, brinous,
P. multocida suppurative bronchopneumonia (n = 3), acute mild to moderate and focally diffuse
M. haemolytica suppurative bronchopneumonia (n = 10) and acute moderate to severe dif- fuse
congestion and pulmonary edema (n = 11). Among these 24 fatal cases, 22 yielded virus
identication and/or bacterial isolation. Eight of these 22 cases were positive for only one
pathogen (5 for viruses and 3 for bacteria), 10/22 were positive for 2 pathogens RSV plus
bacteria (n = 7), PI-3 plus bacteria (n = 2) and 1 for both bacteria and 4/22 were positive for
3 pathogens [RSV, PI-3 plus bacteria (n = 3) and PI-3 plus both bacteria (n = 1)]. Among
the affected lung tissues, virus was identied 19 times (13 positive for RSV, 9 for PI-3, and
3/19 for both viruses) whereas bacteria were isolated 14 times [P. multocida (n = 8), M.
haemolyt- ica (n = 6), and both bacteria (n = 2)]. The presence of multiple pathogens
was observed in 14/22 positive cases with an observation of virusbacteria association
in 13/14 of the cases. The coexistence of RSV-Pm was the most frequently observed
(6/13) followed by the simultaneous presence of RSV-Mh (4/13) and PI-3 Pm or Mh (4/13).
These results appear to indicate that acute pneumonia in alpaca neonates may well result
from virus and bacterial interactions in a similar way to pneumonic lesions of other
ruminants.
2011 Elsevier B.V. All rights reserved.

1. Introduction

Alpaca farming is the most important activity for

Corresponding author at: Departamento de Salud Animal, Facultad
de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos,
Peru. Tel.: +51 1 6197000x5007; fax: +51 1 4375627.
E-mail address: rrosadio@gmail.com (R. Rosadio).
tra- ditional small herders throughout the Peruvian
Andes. Reared at more than 3800 m elevation above
sea level, these animals are the main, and in many
cases the only, source of food and income for a vast
rural population (Bustinza et al., 1988; Wheeler, 1991)
whose herds are

0921-4488/$ see front matter 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.smallrumres.2011.02.001
R. Rosadio et al. / Small Ruminant Research 97 (2011) 110116 111
continuously devastated large coop- erative farm necropsied within 12 h post s
mortem, while 3 were grossly a
by massive neonatal (n = 3) in Southern Peru. m
examined in the eld
mortality produced by Following the results of p
immediately after death and l
infectious diseases, Victorio et al. (2004), we tissues were collected. i
especially enteric and used the direct n
g
acute pneumonia immunouores- cence 2
.
(Rosadio et al., 1990; test to monitor the 2 At the necropsy, individual
Ramrez, 1991). tissues from each . affected lung tissues were
Respiratory infections affected lung for the taken asepti- cally for
in alpacas are believed presence of PI-3 and G microbiology and
r histopathology analysis. First, a
to result from the BRSV antigens. o sterile swab was used to collect
interaction of multiple Additionally, bacte- rial s pulmonary exudates and
factors including pneu- isolation under aerobic s transferred to Stuart trans- port
monic pathogens, conditions on the same media for bacterial analysis
p while a separate sample was
immune status of the tissues revealed massive
a preserved in liquid nitrogen for
host and the extreme predominance of t use in virus antigen detection.
Andean climatic Pasteurella multocida and h The direct uorescent
o
conditions (Rosadio et Mannheimia haemolytica, l
al., 1990). Failure of conrming the o
g
passive immune previously reported y
transfer has been prevalence of these
associ- ated with fatal bacteria in acute e
enteritis or pneumonia pneumonia cases in Peru x
a
in this species (Rosadio et al., 1990). m
(Garmendia et al., 1987), i
however, the denitive 2 n
. a
etiology of most of the t
lower respiratory tract i
M o
infections is still poorly a n
characterized. None the t
e
less, serologic studies r
The postmortem study
have demonstrated that i concentrated on description of
a the type and severity of
llamas and alpacas have lesions found in the
l
been exposed to s pulmonary cavity and
Parainuenza virus 3, regional lymph nodes.
Bovine Diarrhea virus, a Although alpaca lungs lack
n interlobular septa, we
Bovine Her- pesvirus 1,
d described the lesions relative to
Respiratory Sincytyal bronchial branching (Lpez,
virus and Inuenza virus m 1995), recording and char-
A (Rivera et al., 1987; e acterizing gross sessions on
t the cranial, middle (cardiac)
Rosadio et al., 1993) as
h and caudal (diaphragmatic)
well as to Mycoplasma o lobes.
spp. (Hung et al., 1991). d
s
In 2004, Victorio et 2
al. reported the 2 .
. 3
presence of neu- .
1
tralizing antibodies to .
BRSV (80%) and PI3 virus M
(67.5%) in serum samples A i
n c
of 354 apparently healthy r
i
adult alpacas col- lected m o
after a pneumonia a b
l i
outbreak from 30 o
s
different small herders in l
o
Cusco Department. In A total of 24 dead neonates g
the present study, we (crias) aged 20 9 days old (5 y
39 days) and weighing 7 1 kg
report gross,
with a presumptive diagnosis of
microscopy and a
pneumonia and/or hav- ing n
microbiological analysis respiratory difculties were d
of collected from the two La
24 lungs from 7 to 39 Raya Research Stations in
p
Puno (n = 13) and Cuzco (n = 8), a
day old alpacas dying of
as well as from a cooperative t
acute pneu- monia farm (n = 3) in Puno. All the h
collected during the neonates were affected o
l
2004 and 2005 birth suddenly and, with the
o
seasons at exception of 2, did not receive
g
antibiotherapy prior to the y
2 experimental research
fatal out- comes. Most of the
stations (n = 21) and a dead carcasses (n = 21) were
antibody test against (n = 3) or purulent consolidation areas were
parainuenza type 3 (PI3) and necrotic pneumonia (n = noticeable only at the
bovine respiratory syncytial
10) compromis- ing both cut surface.
virus (BRSV) uorescein-
conjugated antisera were used lungs and mainly The extent and
to detect viral antigens, affecting the cranial and grading of the
following the manufacturers ventral areas. The histopathological
protocol (VMRD Inc., Pullman, remaining 11 cases changes matched the
USA). The swabs were plated
showed moderate observed gross lesions
onto 5% sheep blood and
MacConkeys agar for 24 h at conges- tion and and separated into

37 C under aerobic pulmonary edema. 3 different groups: (1)
conditions and the growth of Two of the 3 lungs acute severe and diffuse
the different bacterial species
affected with extensive suppura- tive brinous
was recorded on a semi-
quantitative scale: pure
brinous necrotizing necrotizing
culture, a few bacteria in pneumonia had lesions bronchopneumonia (n =
mixed culture and no growth. compromising the peri- 3); (2) mild to moderate
Bacterial identication was cardic sac and pleura, suppurative diffuse
conducted according to the
with membrane bronchopneumonia (n =
standard protocol described in
Cowan Steels manual (1974) adhering to the rib cage 10) and (3) moderate to
while biochemical testing (API and accumulation of severe and diffuse
20NE, BioMrieux) was used abundant bloody pulmonary congestion
to identify P. multocida and/or brinous exudates and edema (n = 11).
M. haemolytica bacteria. Lastly,
within the thoracic The most severe
representative samples were
xed in 15% formalin, cavity. In the 10 cases histopathological
parafn-embedded, and with purulent necrotic changes were seen in
stained with haematoxylin and pneumonia, gross tissues from the rst
eosin for histopathological lesions affected both group (n = 3) with
evaluation following the
protocol of Zanabria et al.
lungs with evident red lesions extend-
(2000). consolidation
compromising the
3 cranial and middle lobes
. and eventually extending
into the caudal lobes.
R Surface cuts of most of
e these lesions released
s purulent exudates of
u mixed blood and
l mucous, and the lesions
t
s extended into the
bronchial ducts. The
mucosa of both the
3.1.
Gross trachea and bronchi were
and diffusely congested and
histop in 2/10 cases we
atholo observed multiple and
gical overt bullae on the
chang
es parenchyma.
In the remaining
Most of the animals 11/24 affected cases, the
were found dead or lungs were heavier than
dying, and clin- ical normal, displaying a
information of the homogenous red col-
disease process was oration and a shiny-
generally not available. translucent-moist
In one neonate, appearance. The cut
however, we observed surfaces generally
weak- ness, prostration released abundant
with dyspneic frothy uid intermixed
respiration and with bloody exudates
stertoruos sounds at within the affected
auscultation, but no parenchyma and
nasal or ocular secretion adjacent trachea, bronchi
or coughing occurred and bronchiole ducts. In
prior to death. some cases, multiple
In 13/24 of the cases, emphysematous bullae
gross alterations of the were localized on the
lungs were associated caudal lobes. In one case,
with extensive brinous however, multi- ple
necrotizing pneumonia distinct small red
112 R. Rosadio et al. / Small Ruminant Research 97 (2011) 110116
Fig. 1. (a) Lung parenchyma
showing extensive loss of
airspaces with irregular Fig. 2. (a) Lung tissue showing
necrotic foci (arrows) diffuse congestion and alveoli
intermixed with congestion; containing edematous uid
H&E, 5. (b) Focally extensive and/or mixed inammatory
coagulative necrosis with cells; H&E, 10. (b) Focal
degenerate lung cells and congestion with mild
multiple bacterial nodules
polymorph inltration within
(arrows); H&E, 40.
the bronchial sub- mucosa
(arrows), and mild bronchi
ing into and hyperplasia with mixed debris
and inammatory cells within
compromising the
the bronchial lumen (arrow
pleura, producing heads); H&E, 40.
diffuse and extensive
loss of normal tissue
pleural compromise. The
architecture with the
least damaged tissue
presence of irregular
exhibited minimal
masses of pink to
neutrophil inltrations
violet necrotic areas
mixed with
(Fig. 1a) and multiple
proteinaceuos material
foci of coagulative
in the alveolar spaces,
necro- sis intermixed
with mild congestion
with bacteria colonies
and epithelial
and inltrated with
hyperplasia (Fig. 3a).
inammatory cells,
The more affected cases
mainly polymorphs (Fig.
showed degeneration of
1b). Addition- ally, areas
the alveolar epithelial
of abundant mixed
cells, with sloughed
exudates of polymorph
bronchial and
and mononuclear cells
bronchiolar cells
were observed, with
intermixed with
eventual intra- alveolar
neutrophil deposition
brin deposition and
intraluminally and
presence of red blood
eventually extending
cells (Fig. 2a). The
into the sub mucosa (Fig.
adjacent bronchi and
3b).
bronchioles contained
Lesions within the
exudates of mixed
third group (n = 11)
inammatory cells and
generally involved the
cellular debris,
vascular bed with
obliterating the lumen
minimal inammatory
(Fig. 2b). In 2 cases, the
component. Small blood
blood vessels were
vessel plethorization
plethorized and
and pres- ence of
congested within the
proteinaceous uids
interalveolar septa with
within the alveolar
focal thrombus
spaces (Fig. 4a) were
formation. In all 3
commonly observed,
cases the pleura was
occasionally com-
thickened with
promising both the
lamentous brin
bronchiolar lumen and
deposition, polymorph
connective sub-pleural
cells, with evidence of
tissue with the presence
blood and lymphatic
of a mild multifocal
vessel congestion and
neutrophil inltration of
foci of brinous and/or
the alveolar spaces and
suppurative exudates.
adjacent interstitial
The lesions in the
tissues, and moderate
second group (n = 10)
hyperplasia of the bron-
corresponded to mild to
chiolar epithelial cells
moderate
(Fig. 4b). Occasionally,
bronchopneumonia with
deposits of
minimal
R. Rosadio et al. / Small Ruminant Research 97 (2011) 110116 113
 suppurative Fig. 4. (a) Severe pulmonary
Fig. 3. (a) Severe pulmonary bronchopneumonia congestion intermixed with
congestion with abundant brin, inam- matory
lesions (Table 1).
polymorph cells within the polymorph cells (arrow heads)
alveolar spaces and bronchiolar and macrophages (arrows)
cell hyperplasia; H&E, 40. (b) within the alveolar spaces;
Lung with massive airspace H&E, 40. (b) Mild
losses caused by extensive pulmonary congestion with
congestion and mixed moderate mixed inammatory
inammatory cells with cell inltration and
deposition of proteinaceus degeneration of the epithelial
materials and inammatory bronchiolar cells; HE, 40.
cells within bronchiolar lumen;
H&E, 40.
Bacteria were
cultivated from 16
hemosiderin were (72.7%) lung tissues,
observed within the with Pasteurella (n = 11)
alveolar spaces, and and Mannheimia (n = 7)
multiple isolated in high numbers
emphysematous areas in both pure (n = 14) and
associated with ruptured mixed cultures (n = 2)
alveolar cells were seen. (Table 2). Colonies of M.
haemolytica were small
3.2. Viral and grey, with a narrow
antigen zone of haemolysis after
detection and 1824 h of incubation. In
bacteria
isolation the API system, all the
isolates were indole
In 22/24 cases, the negative and fermented
presence of virus arabinose. Echerichia coli
antigen was detected (n = 2) and yeast (n = 1)
and/or bacteria were were also isolated but
isolated. In 8 of the 22 were not considered
ani- mals a single agent pathogenic. P. multocida
(virus or bacteria) was was primarily found in
identied, while in the lesions classied in the
remaining 14, two or second (n = 6) and third
three agents (virus (n = 4) groups, while
coexisting with bacteria) only one case was found
were detected. The 2 in a group 1 animal with
negative animals had severe histopathological
gross lesions of lesions (Table 2). In
pulmonary congestion contrast, M. haemolytica
and edema. (n = 7) was recovered
The direct IF test from animals distributed
detected a total of 19/22 in all 3 groups (Table 2).
viral antigens with It is important to note
predominance of BRSV (n that 2 of the 3 negative
= 13), followed by PI3 cases pertained to
virus (n = 9) and animals that received
presence of both viruses antibiotherapy prior to
in 3 cases. Eight of the sampling.
13 BRSV-positive cases,
had histopathological 3.3. Association
lesions clas- sied as between pathogens
mild to moderate and histopathology
suppurative features
bronchopneumonia
(Group 2) and 5/13 In 63.6% of the cases
moderate to severe (14/22) we detected
pulmonary conges- tion multiple infec- tions
and edema (Group 3). Of (virus and bacteria),
the nine PI3-positive including ten with dual
cases, 4 had moderate to infections and four with
severe pulmonary triple agents. In the
congestion, 5 had mild dual infections, the
to moderate
bronchopneumonia (n =
3) and 2 had extensive
114 R. Rosadio et al. / Small Ruminant Research 97 (2011) 110116

Table 1
Association between histopathology, viral identication and/or bacteria isolation.

Type of lesion BRSV PI-3 Pasteurella multocida Mannhemia haemolytica

 Group 1. Extensive suppurative and 0 2 1 2
severe brinous necrotizing
bronchopneumonia (n = 3)
Group 2. Mild to moderate suppurative 8 3 4 3
bronchopneumonia (n = 10)
Group 3. Moderate to severe 5 4 6 2
pulmonary congestion and edema
(n = 11)
Total 13 9 11 7

association between BRSV and P. multocida was the most
frequently observed (n = 4) followed by BRSV-M.
haemolyt- ica (n = 3), PI-3 with P. multocida (n = 2) and the
BRSV-PI-3 (n = 1). In the case of triple infections (4/22),
the interaction between BRSV- PI-3 and P. multocida was
observed twice, while the association between BRSV-P.
multocida and M. haemolytica, as well as PI-3-P. multocida
and M. haemolyt- ica were both observed once (Table 2).
In the remaining 8 positive cases, the participation of a
single agent PI-3 was observed 3 times, BRS twice, M.
haemolytica twice and P. multocida once (Table 2).
4. Discussion
Acute pneumonia is the second cause of mortality
among Andean alpaca neonates, but the denitive
etiologic agent has not been determined. The results of
this study show the presence of pneumotropic viruses
(BRSV and PI3), in conjunction with P. multocida and/or
M. haemolyt- ica, in a range of pathological pneumonic
lesions in alpaca neonates, supporting preliminary
serological evidence for the presence of multiple agents
within acute alpaca pneu- monias in Peru (Rivera et al.,
1987; Hung et al., 1991; Rosadio et al., 1993; Victorio et
al., 2004).
The gross changes observed in the 24 pneumonia
cases showed different degrees of pathology varying from
congestion and pulmonary edema (Group 3, 11/24), bilat-
eral purulent pneumonia (Group 2, 10/24) to severe and
extensive necrotizing purulent pneumonia with brinous
pleuritis (Group 1, 3/24). The more severe lesions
compro- mised both lungs, principally involving the
anterior ventral aspects of the cranial lobes with
hemorrhagic brinous pleuritis. These types of gross
lesions have been described as typical of acute
pneumonias in alpacas (Rosadio et al.,
1990) and are consistent with features attributed to
bovine lobar pneumonia or brinous bronchopneumonia
associ- ated with multiple agents, including Pasteurella
spp. of calves, and small ruminants (Dungworth, 1993;
Piojan et al., 1999; Donachie, 2007).
The gross changes found among the 10/24 animals
with bilateral purulent pneumonia (Group 2) were
mostly red consolidations affecting cranial and ventral
areas of the lungs, with multiple abscesses and, in one
case, pleuritis. These pathologies bear some
similarities to suppura- tive bronchopneumonia in
cattle (Dungworth, 1993). In alpacas, the gross
alterations observed in animals suffering from congestion
and pulmonary edema, must be evidence of an early
inammatory process within the lungs, in con-

Table 2
Association between histopathology lesions and pathogens.

Type of lesion Agents Pathogens N

Group 1. Extensive suppurative 3 PI-3, Mh, Pm 1

and severe brinous necrotizing 1 PI-3 1
bronchopneumonia (n = 3) Mh 1
Group 2. Mild to moderate 3 BRSV, PI-3, Pm 1
suppurative bronchopneumonia 2 BRSV, Pm 3
(n = 10) BRSV, Mh 2
BRSV, PI-3 1
1 BRSV 1
PI-3 1
Mh 1

Group 3. Moderate 3 BRSV, Pm, Mh 1

to severe pulmonary BRSV, PI-3,Pm 1
congestion and 2 PI-3,Pm 2
edema (n = 11) BRSV, Pm 1
BRSV, Mh 1
1 PI-3 1
Pm 1
1 BRSV 1
0 negative 2

Total 24

BRSV, Bovine respiratory syncytial virus, PI-3, Parainuenza type 3 virus; Pm, Pasteurella multocida; Mh, Mannheimia haemolytica.
R. Rosadio et al. / Small Ruminant Research 97 (2011) 110116 115
trast to those described alveolar bedding multocida (n = 4) and M. were detected in all
(enlargement, redness contains inamma- tory haemolytica (n = 3). In the three groups, including
and shiny surface) of the exudates which triple associations (n = the least affected group
early stages of eventually obliterate 4), the presence of 3 where 8/11 animals
pneumonia seen in the bronchi, bronchiolar BRSV was the most suffering from pul-
calves and lambs (Lpez, and alveolar lumens. All frequent (n = 3) monary congestion and
1995; Kimberling et al., of these changes are combined with PI3 and edema lesions
1988; Martin, commonly observed and P. multocida (n = 2), as registered viral
1 described in aspiration well as with both infections. The presence
9 pneumo- nia with bacteria (n = 1) (Table 2). of virus infections at the
9 participation of bacterial The presence of dual and initial stage of the
6 triple infections
infections (Lpez, 1995). process suggests that
)
The common associated with BRSV they may be a triggering
.
histopathology features was most frequently factor in the development
The histopathology
in this group are observed in animals of acute pneumonia. It is
lesions tend to
degrees of vascular from the second and possible that the
correlate gross
damage, inammatory third groups (Table 2). destruction of pulmonary
pathology with the
cell exudates and The identication of PI-3 epithelial cells observed
more dramatic
extensive necrosis of virus was observed in 3/8 in these 8/11 cases may
alterations found in
bronchial and/or single infections, 2/10 be the product of viral
those animals affected
bronchiolar cells with dual and in 1/4 of triple replica- tion and may
with severe necrotizing
focally extensive infections. explain the edema,
bron- chopneumonia
epithelial denudation and The three groups hemorrhages and hyaline
and pleuritis (Group 1).
exu- dates and intra- with differing degrees membranes as suggested
Similar features have
luminal debris of pathology found in this by Bryson et al. (1979)
been described as
accumulation (Fig. 3a and study reect variations and con- sistently seen in
brinous
b). These alterations have of possible complex most of the affected
bronchopneumonia in
been described to be a pathogen interactions for animals within this
small ruminants, as
consequence of viral inducing disease. Viral group.
well as
replication (Bryson et al., antigens BRSV was frequently
pleuropneumonia in
1983; Davies et al., 1981; detected (8/10) within
cattle (Dungworth,
Domachowske and the group
1993; Lpez, 1995). The
Rosenberg, 1999). 2 cases (Table 2) and
changes found in this
The identication mostly seen in multiple
rst group reect a
and/or isolation of infections (n = 7)
mixed and progressive
multiple microor- suggesting a potential
inamma- tory process
ganisms in individual cause of epithelial
in which the initial
cases indicates that, as necrosis and hyperplasia
changes are associated
is the case in other which could easily
with vascular
ruminants, acute predispose to bacterial
congestion and
pneumonia in alpacas, colonization as
presence of abundant
may well result from suggested by Masot et al.
b- rinous exudates
the interaction between (1996). The ele- vated
within the alveolar
virus and bacteria presence of BRSV in dual
spaces, interalveolar
(Davies et al., 1981; and triple infections, in
septae and extending to
Kimberling et al., 1988; this group, and mainly
the pleura. Whereas, the
Martin, 1996). In 91.7% of with P. multocida (n = 4),
presence of massive and
the 24 cases, the presence M. haemolyt- ica (n = 2)
extensive diffuse
of potential pathogens or virus PI-3 (n = 2),
coagulative necrosis
within pneumonic indicates that BSRV could
with multiple bacterial
lesions was detected, be one of the main
accumulations and
and in more than half pneumonia-inducing
mixed inammatory
(14/22) the association agents. This is sup-
responses are presumably
between virus and ported by the high
later changes (Fig. 1a and
bacteria was evident. In prevalence of specic
b).
10/14 of these cases, 2 antibodies to BRSV found
The moderate
pathogens were in communal and
suppurative
coexisting in the cooperative alpaca farms
bronchopneumonia
affected lung and, in in Peru (Manchego et al.,
lesions found in 10
the remaining 4, three 1998; Victorio et al.,
grossly affected alpacas
microorganisms were 2004). It is worth
from Group 2 resem- ble
present (Table 2). Among mentioning that PI-3
those described as
the dual associations, the virus was present in
suppurative pneumonia
combination between 2/3 affected cases of
in other ruminants
BRSV/bacteria (n = 7) severe brinous
(Dungworth, 1993;
was the most bronchopneumonia
Donachie, 2007) where
frequently observed, (Group 1, Table 1) and
the bronchiolar and the
preferentially with P. associated with both
bacteria (M. haemolyt- conrms previous
ica and P. multocida) in identication of P.
one of the cases. This multocida, and P.
virus has also been (Mannheimia)
implicated as a haemolytica from fatal
predisposing factor in acute pneumonia in
producing severe neonates (Ramrez, 1991;
bronchopneumonia, Rosadio et al., 1990).
principally in Although we have
conjunction with M. not elucidated
haemolytica in lambs environmental fac- tors,
(Davies et al., 1986; the climate must play a
Kimberling et al., 1988). role, since the birth
The PI-3 virus has been season coincides with
reported to induce the wet season, which is
degenerative necrotic intermixed with short
changes of alveolar and little summer
bronchio- lar epithelial (veranillo) periods
cells, as well as characterized by lack of
bronchiolar hyperplasia rainfall and extreme
with syncytia formations daily temperature vari-
(Bryson et al., 1979; ation (greater than >20

Davies et al., 1981; Yener C) (Winterhalder and
et al., 2005; Intisar et al., Thomas,
2010). In the present
study we observed the
rst two changes in
2/3 of animals in Group
1.
The isolation of P.
multocida and M.
haemolytica in pneu-
monic cases tends to
conrm that these
pathogens are major
contributors to acute
respiratory diseases
(Hirsh and Biberstein,
2004). M. haemolytica
can cause severe pneu-
monia in cattle and
sheep (Yates et al.,
1983; Ackermann and
Brogden, 2000) and has
been isolated from
laryngeal abscessation in
an alpaca (Dwan et al.,
2008). It has been
depicted as a crucial
factor inducing brinous
pneumonia in most
animals exposed to
stressor factors or
following viral infections
(Davies et al., 1981,
1986). The presence of
M. haemolytica in 7
cases from the 3 groups,
and particu- larly in 2/3
cases of lung lesions
with necrotizing
brinous pleuritis, tends
to support this
pathogenicity capacity.
The isolation of
Pasteurella and
Mannheimia in
pneumonic lesions
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