Vet Pathol 45:5153 (2008)

A Clonal Outbreak of Acute Fatal Hemorrhagic Pneumonia in Intensively Housed (Shelter)

Dogs Caused by Streptococcus equi subsp. zooepidemicus
P. A. PESAVENTO, K. F. HURLEY, M. J. BANNASCH, S. ARTIUSHIN,

AND

J. F. TIMONEY

School of Veterinary Medicine, Department of Pathology, Microbiology, and Immunology (PAP) and
Koret Shelter Medicine Program (MJB, KFH), University of California at Davis, Davis, CA;
and Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky,
Lexington, KY (SA, JFT)
Abstract. An outbreak of acute, fatal, hemorrhagic pneumonia was observed in more than 1,000
mixed breed dogs in a single animal shelter. The Department of Anatomic Pathology at the University of
California at Davis School of Veterinary Medicine performed necropsies on dogs that were found
moribund in acute respiratory distress or found dead with evidence of nasal bleeding. All dogs had
hemothorax and an acute, fibrinosuppurative pneumonia. Large numbers of gram-positive cocci were
observed within the lungs of all dogs and within septic thromboemboli of remote organs in about 50% of
cases. Bacterial cultures from the dogs and their environment revealed widespread beta-hemolytic
Streptococus equi subspecies zooepidemicus (Lancefield Group C). Extensive diagnostic testing failed to
reveal the consistent presence of copathogens in individual cases. The clinical, epidemiologic, molecular
biologic, and pathologic data indicate that a single clone of S. zooepidemicus was the cause of an acutely
fatal respiratory infection in these dogs.
Key words:

Canines; pneumonia; respiratory disease; Streptococcus; SzP protein.

service at the School of Veterinary Medicine at UC
Davis performed necropsies on 8 dogs found dead or
acutely moribund and euthanized.
On gross examination all dogs had hemothorax (200
900 milliliters) and rubbery, mottled dark to bright red
lungs (Fig. 1). Variably sized and poorly demarcated
areas of collapse were present in all lung lobes. Three of
the animals had a concurrent acute necrotizing rhinitis
and sinusitis. Histologically, there was acute to peracute
fibrinosuppurative pneumonia with extensive to lobular
obliteration of alveolar spaces with neutrophils (Fig. 2).
Large and medium-sized airways were unaffected, but
inflammation was intense around most terminal airways. Occasional small vessels were thrombosed with
fibrin-enmeshed erythrocytes and leukocytes. Variable
to large numbers of intralesional gram positive cocci
were present within the cytoplasm of macrophages and
free within the extracellular space in pairs, strings, and
large colonies (Fig. 3 and inset). Septic thromboemboli
were less consistently observed (3/8) in the spleen and
renal glomeruli (Fig. 4). No evidence was detected of
prior (chronic) lung inflammation or bronchiolar injury
on histologic examination of these cases.
Polymerase chain reaction (PCR)-based amplification
of common canine respiratory pathogens was performed
on the lung tissue of 5 of 8 necropsied dogs (Table 1).
Canine adenovirus, canine herpesvirus, and distempervirus were inconsistently detected. Canine influenza
virus, canine parainfluenza virus, and B. brochiseptica
were undetectable by this method. Under the aegis of
the Koret Shelter Medicine Program, similar panels
were performed on oropharyngeal samples from dogs
with respiratory disease at this shelter in 2003 and 2005.

Canine infectious respiratory disease (CIRD, kennel

cough) is a clinical syndrome nearly ubiquitous in
animal shelters and other densely housed canine
populations. Clinical signs of disease last days to weeks,
and, although death is a rare sequela to disease,
intractable and even mild respiratory disease can be
a criterion for euthanasia in some shelters. The etiology
of CIRD is multifactorial, and host and environmental
factors such as stress and crowded conditions are likely
to contribute to morbidity. In the past, the most
commonly associated bacterial agent was Bordetella
bronchiseptica.4 However, as a result of some combination of enhanced detection, sampling site selection,
vaccination, and/or natural evolution, other bacterial
agents have been recently or increasingly implicated.2,3,5
In one study, isolation of S. equi subsp. zooepidemicus
from the respiratory tract of both healthy and diseased
dogs increased dramatically with increasing clinical
respiratory disease.2
In a single shelter with an intake of ,50,000 animals/
year, CIRD was an ongoing problem of high morbidity
but low mortality. In the summer of 2006, veterinarians
at the shelter noticed an increasing number of dogs
found acutely moribund and in respiratory distress or
dead with bleeding from the nose and/or mouth. In
February 2007, more than a thousand dogs were
estimated to have suffered or died from severe hemorrhagic pneumonia. Dogs with low-grade respiratory
disease were not included in the estimate. The Humane
Society of the United States and members of the Koret
Shelter Medicine Program at the University of California at Davis inspected the facility in February of 2007.
During the 2-week inspection period, the necropsy
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Brief Communications and Case Reports

Vet Pathol 45:1, 2008

Fig. 1. Thoracic cavity, canine; Case 1. The space between the lung (asterix) and the diaphragm in the caudal
and dorsal thoracic cavity is filled with unclotted, hemmorrhagic fluid.
Fig. 2. Lung; canine; Case 1. Alveolar septae are obscured, and alveolar spaces are filled with neutrophils and
fibrin. HE. Bar 5 100 mm.
Fig. 3. Lung; Case 1. Numerous chains and clusters of gram-positive cocci are free within alveolar spaces and
engulfed by alveolar macrophages (inset, arrows). Brown and Brenn Gram stain.
Fig. 4. Spleen; Case 3. A large cluster of coccoid bacteria distends a sinusoid within the red pulp. HE. Bar 5
50 mm.
At those times, hemorrhagic pneumonia was not
present, death was uncommon, and B. bronchiseptica
but not S. zooepidemicus was commonly detected in
dogs with respiratory disease.
Lung tissue from 6 of 8 necropsied animals was
submitted for culture to the Microbiology Service, VMTH,
UC Davis. Two samples from the environment (sink, cage
floor), 2 samples from the upper respiratory tract of
neighboring dogs (oropharynx), and 1 sample from a foot

abscess of an affected dog were also submitted. S. equi

subsp. zooepidemicus was cultured from all samples. In 3 of
6 samples cultured from lung and from the foot abscess, S.
zooepidemicus was the sole bacterial isolate. Bacterial
identification was based on observation of gram-positive,
catalase-negative, chain-forming cocci, which fermented
lactose and sorbitol but not trehalose. The bacteria were
not susceptible to doxycycline, the antibiotic most commonly used in the affected shelter. Each bacterial isolate

Table 1. Prevalence* of common pathogens{ in dogs with respiratory disease (2003, 2005) and in cases of fatal
hemorrhagic pneumonia (2007).

2003{
2005{
20071

Bordetella

CHV

CAV-2

CDV

CPI

CI

16/17
10/20
0/5

9/17
15/20
1/5

0/17
4/20
3/5

1/17
1/20
1/5

0/17
0/20
0/5

0/5

* Number of dogs affected/number of dogs examined.

{ CHV 5 canine herpesvirus; CAV-2 5 canine adenovirus type II; CDV 5 canine distemper virus; CPI 5 canine parainfluenza
virus; CI 5 canine influenza virus.
{ Oropharyngeal swab.
1 Lung (necropsy).

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Brief Communications and Case Reports

Fig. 5. Polymerase chain reaction amplification of

DNA fragments from S. zooepidemicus isolated from
dogs and the shelter environment using primers FUS
and FDS located upstream and downstream, respectively, of se18.9.7 Lanes 212 are fragments amplified
from colonies isolated during the time of the outbreak.
Lanes 6, 8, and 11, lung (cases 13); lane 10, foot abscess
(case 4); lanes 5 and 7, oral swabs of neighboring dogs;
lane 4, 9 environmental isolates (sink, cage floor); lane 3,
spleen from a cat that died acutely with respiratory
failure during the canine respiratory outbreak (lane 2 is
from an isolate from the nasal cavity of this cat). Lane 1
shows a fragment amplified from an isolate of a dog that
died 2 months after the outbreak had ended.
was also screened by PCR for Se18.9,7 including its
flanking sequences, and by immunoblot of mutanolysinextracted proteins for reactivity with rabbit antiserum to
recSzPW60,6,8 the Moore and Bryans typing antigen of S.
zooepidemicus. All isolates from dogs at the time of the
outbreak (lanes 311, Fig. 5) yielded a similar-sized
amplicon by PCR (Fig. 5). This result and the observed
similarity in size of the SzP proteins (data not shown)
indicated that the shelter outbreak of hemorrhagic
streptococcal pneumonia was caused by a single clone of
S. zooepidemicus and that the environment was contaminated with the same clone. S. zooepidemicus was also
cultured from the spleen of a cat that died during the canine
epizootic and from the lung of a dog that died 2 months
after the outbreak had ended. However, these isolates
yielded 2 different amplicons distinct from both the
outbreak isolate and each other (lanes 12, Fig. 5).
In horses, S. zooepidemicus is a commonly isolated
mucosal commensal that opportunistically invades as
a sequela to virus infection, high ambient temperature, or
prolonged transportation. Infections are endogenous in
that a preexisting clone in tonsillar tissue is selected, is
amplified, and eventually dominates in the lower respiratory tract.1,6 Unlike the shelter epizootic described
here, the clone found in pneumonic lungs varies from
horse to horse. Dominance of a single clone of an
organism only infrequently detected in healthy household
dogs indicates that environmental conditions in combination with stress and possibly other as yet unknown
microbial agents provided a setting for rapid invasion
into densely populated and highly susceptible hosts.
Many of the cases described in the shelter were in dogs
admitted to the environment as early as 2 days prior to

their death. The short interval between entry to the

kennel and onset of fatal pneumonia suggests either
a challenge that was overwhelming or selection of a clone
with enhanced virulence. In this outbreak, the most
common canine respiratory copathogens either were
absent or were inconsistently present. In addition, there
was no histologic evidence of predisposing viral damage,
which was consistent with the death of several dogs
within 2 days of admission to the shelter. In a previous
report, an outbreak of hemorrhagic pneumonia in
kenneled research dogs was attributed to S. zooepidemicus.3 Although the outbreak described in this paper was
more protracted and involved more animals, the pathologic and microbiologic features were otherwise similar.
The outbreak ended subsequent to imposed husbandry changes, including a dramatic depopulation, facility
cleaning, and staff education. Attempts to identify
potential virulence factors in the clone from this shelter,
including antiphagocytic proteins, proteases, and pyrogenic exotoxins, are ongoing.

References
1 Anzai T, Walker JA, Blair MB, Chambers TM,
Timoney JF: Comparison of the phenotypes of
Streptococcus zooepidemicus isolated from tonsils of
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162166, 2000
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Request reprints from Dr. Patricia A. Pesavento,
University of California, Davis School of Veterinary
Medicine, PMI Room 4206, VM3A One Shields Avenue,
Davis, CA 95616-5270 (USA). E-mail: papesavento@
ucdavis.edu.

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