Professional Documents
Culture Documents
RABBİT-Pasteurellosis in Rabbits
RABBİT-Pasteurellosis in Rabbits
4 April 2000
Pasteurellosis in Rabbits
University of Georgia
FOCAL POINT Susan Sanchez, PhD Shaikh Mizan, PhD
Branson W. Ritchie, DVM, PhD Margie D. Lee, DVM, PhD
★Serology combined with
polymerase chain reaction–based
assays can be used to rapidly ABSTRACT: Pasteurella multocida infections in rabbits may remain subclinical or can cause a
and accurately document variety of acute or chronic diseases in most organs, particularly tissue in the repiratory and
pasteurellosis in rabbits. genital tracts. Although clinical signs are frequently used to diagnose pasteurellosis, other or-
ganisms can cause similar clinical signs. The status of a rabbit with respect to P. multocida
can be best determined using a polymerase chain reaction–based assay in conjunction with a
sensitive and specific ELISA.
KEY FACTS
P
asteurella multocida can cause various clinical changes in rabbits. As one
■ Pasteurella multocida is one of of the most common disease-causing agents in this genus, P. multocida
the most common disease- has been associated with rhinitis (snuffles), abscessation, pneumonia, oti-
causing agents in rabbits. tis, conjunctivitis, genital tract infections, abortions, neonatal mortality, and sep-
ticemia.1,2 Although this organism is highly infectious, many rabbits are subclini-
■ Culture accuracy of P. multocida cally infected until stress or concomitant problems precipitate recognizable
depends on the transport disease. The reported prevalence of P. multocida in clinically stable rabbits ranges
medium, storage temperature from 20% to 90%, depending on the animal’s age, sex, and health status and the
of the swab, previous antibiotic chosen testing method.3,4
treatment, and laboratory culture Based on studies by Webster and Smith, it is generally believed that rabbits in-
conditions. fected with P. multocida can resist or clear infection, become chronically but sub-
clinically infected, or develop acute or chronic disease. The type of infection is
■ A polymerase chain reaction–based believed to be influenced by the strain of infecting Pasteurella and the specific
assay is a more sensitive test than host resistant to that strain.5 Generally, Pasteurellaceae should be considered
is culture, partially because the commensal organisms that may be pathogenic in immunosuppressed rabbits.
former detects the presence of Peracute pasteurellosis usually causes septicemia and death in seemingly
both viable and nonviable bacteria. healthy rabbits. Upper respiratory tract disease (e.g., snuffles) associated with P.
multocida infection starts as a serous nasal discharge that usually turns mucopu-
■ Serology can be used to detect P. rulent (Figure 1). Hair on the animal’s forelimbs is usually moist from repeated
multocida antibodies, particularly in attempts to clean the muzzle. Epiphora, sneezing, and coughing may accompany
chronically infected rabbits. the upper respiratory tract form of pasteurellosis. Anorexia, weight loss, dyspnea,
pleuropneumonia, and, in the most severe cases, death may occur if untreated
■ Nasopharyngeal swabs are rhinitis progresses to more generalized disease.
recommended for obtaining culture P. multocida is a common cause of abscessation in rabbits, irrespective of the
samples to detect the presence of presence or absence of respiratory tract disease. These abscesses are filled with
P. multocida; however, sample thick, yellow–gray mucopurulent exudate and can occur in any location on or
collection from conscious rabbits within the body (Figures 2 and 3). Microscopic examination of material collected
is difficult. from these abscesses usually reveals small gram-negative coccobacilli suggestive of
P. multocida infection (Figure 4). P. multocida can also cause otitis externa and oti-
tis media. If the vestibular system is involved, torticollis with the head tilted to
Compendium April 2000 Small Animal/Exotics
Figure 1—Mucopurulent rhinitis suggestive of progressive Figure 2—Pasteurella multocida infection can cause abscessa-
pasteurellosis. tion in any organ. In this rabbit, the scrotum is ulcerated and
its necrotic abscessed testicle was surgically removed.
DIAGNOSIS
Suggestive clinical signs are frequently used to diag-
nose pasteurellosis. However, other organisms alone or
in combination with viruses and parasites can cause
clinical signs that are characteristic of pasteurellosis.5
Accurate diagnosis and treatment of pasteurellosis are
important to reduce the spread of P. multocida infec-
tion, particularly in multiple-rabbit households or rab-
bitries. Isolating the organism from the rabbit’s nostrils,
conjunctiva, ears, or abscessed tissue has been the most
common method for confirming pasteurellosis. Figure 3—Mucopurulent, necrotizing pneumonia in a rabbit
In a classic study5 involving a rabbit colony with en- with chronic pasteurellosis. The affected lung was surgically
demic pasteurellosis, 58 of 100 rabbits had gross upper removed.
respiratory changes at necropsy. P. multocida was recovered
from the nares of 55 of these rabbits and from 8 rabbits
with no signs of upper respiratory disease.5 However, cul- ture process can produce false-negative results. Immediate
ture of P. multocida in living rabbits can be problematic transfer of samples to culture medium is not always possi-
because even deep nasal cultures frequently fail to confirm ble, and transport medium must preserve the organism
upper respiratory tract infection when an organism in sub- until samples reach the laboratory. Compounding this
clinically infected rabbits is present in low numbers. Local- problem is the fact that P. multocida is extremely delicate
ized infection in the tympanic bullae, caudal oropharynx, and does not survive well in most common transport me-
genital tract, and other internal organs renders ante- dia (e.g. Stuart’s, Amies), in some instances surviving less
mortem sampling for culture difficult.7,8 In other cases, P. than 24 hours at room temperature.9 The organism may
multocida cannot be cultured from obviously diseased or- survive for several days in Cary-Blair medium. Because re-
gans because the rabbit has received antibiotics. frigeration can reduce survival of P. multocida,9 samples for
In addition to problems with sample collection, the cul- culture should be maintained at room temperature or
Figure 4—Pasteurella species are gram-negative coccobacilli Figure 5—Torticollis secondary to Pasteurella-associated otitis
that can frequently be detected in cytologic preparations of media/interna.
secretions or exudates.
TABLE I TABLE II
Summary of the Test Results Results of Serology and Culture
to Diagnosis Pasteurellosis Testing in 52 Rabbits with Clinical
Total Signs Suggestive of Pasteurellosis
Tested Positive Negative No. of Rabbits Serology Culture
16 (46%) – – 27 (71%) – –
10 (29%) + + 9 (24%) – +
8 (23%) – + 2 (5%) + +
1 (3%) + –
TABLE V than half of the sera submitted (30 of 55) tested nega-
tive for anti-Pasteurella IgG antibodies by ELISA.
Results of Culture, Serology, and
Polymerase Chain Reaction Testing in 35 Rabbits with Although samples were submitted for rabbits with
Clinical Signs Suggestive of Pasteurellosis clinical signs suggestive of pasteurellosis, the results of
this study suggest that many cases of suspected pas-
No. of Polymerase
teurellosis may be caused by other organisms alone or
Rabbits Culture Serology Chain Reaction
in combination. Candidate organisms include Bordetel-
16 (46%) – – – la bronchiseptica, Pseudomonas species, Staphylococcus
species or other Pasteurella species.5 Thus although P.
2 (6%) + + + multocida is a ubiquitous bacteria, it may be an over-
8 (23%) – + + stated cause of respiratory disease in rabbits when clini-
cal signs alone are used for diagnosis. The clinical trial
8 (23%) – + –
reviewed in this manuscript suggests that PCR testing
1 (3%) – – + in combination with serology using a neuraminidase–
based ELISA can help confirm pasteurellosis as the
cause of disease in rabbits with suspicious clinical signs.
P. multocida was detected by culture and/or PCR assay
also tested positive on a recently developed ELISA.14 Fur-
thermore, positive serology results indicated that eight ACKNOWLEDGMENTS
rabbits (23%) had been infected with P. multocida in This research was supported by the University of
their immunologically detectable past even though the Georgia Research Foundation Animal Health Fund and
organism was not demonstrated on are nasal swabs for the Veterinary Medical Experiment Station. The au-
PCR assay or culture. Most of the submitted swabs were thors thank Drs. William Chavez, Joe Deck, Cheryl
obtained from the nasal passages, which may have re- Greenacre, Heather McClure, Masako Mori, Robert Ness,
duced the number of bacteria available for detection on Richard Nye, Robert Owen, Pamela Slack, Samuel
culture or PCR assay. P. multocida has an affinity for the Vaughn, and Heather Wilson for submitting samples
mucosal epithelium of the nasopharynx.8,15 Nasopharyn- used in this project.
geal swabs are reportedly 36% more sensitive than are
nasal swabs in detecting P. multocida; however, collecting
a sample from the nasopharynx is difficult in a conscious REFERENCES
1. Percy DH, Barthold SW: Pathology of Laboratory Rodents
rabbit.8 The use of nasal swabs in this study, instability of and Rabbits. Ames, IA, Iowa State University Press, 1993, pp
P. multocida during shipment, and ability of PCR assay 179–224.
to detect nucleic acid from inactive organisms likely con- 2. Dhillon AS, Andrews DK: Abortions, still births, and infant
tributed to the higher sensitivity of PCR assay compared mortality in a commercial rabbitry. J Appl Rabbit Res 5:97–
with that of culture. To screen for bacteria in rabbits 98, 1982.
3. Lu YS, Ringler DH, Parks, JS: Characterization of Pasteurel-
with rhinitis, a No. 4 calcium alginate swab can be in- la multocida isolates from the nares of healthy rabbits and
serted 1 to 4 cm into the nares along the nasal septum rabbits with pneumonia. Lab Anim Sci 28:691–697, 1978.
on both sides.5 4. Weisbroth SH, Scher S: The establishment of a specific-
The ELISA used in this study detected anti-Pasteurella pathogen-free rabbit breeding colony. II. Monitoring for dis-
IgG in 25 of 55 or 45.5% of the sera samples tested. By ease and health statistics. Lab Anim Care 19:795–799, 1969.
5. Deeb BJ: Respiratory disease and the Pasteurella complex, in
comparison, only 11 of 38 rabbits (29%) had positive Hillyer EV, Quesenberry KE (eds): Ferrets, Rabbits and Ro-
results on PCR assay, and only two of 55 rabbits dents. Clinical Medicine and Surgery. Philadelphia, WB Saun-
(3.64%) had positive culture results. All except one of ders Co, 1997, pp 189–201.
the rabbits with positive results on PCR assay and/or 6. DeLong D, Manning PJ: Bacterial diseases, in Manning PJ,
culture also had positive results on serology. Because no Ringler DH, Newcomer CE (eds):The Biology of the Labora-
tory Rabbit, ed 2. San Diego, Academic Press, 1994, pp
additional sample was available from the one rabbit 131–170.
with positive PCR results and negative serology results, 7. Holmes HT, Matsumoto M, Patton NM, Zehfus BR: Sero-
whether the serology was a false-negative reaction, the logic methods for detection of Pasteurella multocida infec-
swab was contaminated with target nucleic acid, or the tions in nasal culture negative rabbits. Lab Anim Sci 36:640–
rabbit had an early infection to which the immune sys- 645, 1986.
8. Holmes HT, Matsumoto M, Patton NM, Harris DJ: A
tem had not mounted a detectable immune-response method for culturing the nasopharyngeal area of rabbits. Lab
cannot be determined. Serology detected more positive Anim 21:353–355, 1987.
results than did any of the other methods, but more 9. Kawamoto E, Sawada T, Maruyama T: Evaluation of trans-
port media for Pasteurella multocida isolates from rabbit Pasteurella multocida to rabbit pharyngeal cells and its possi-
nasal specimens. J Clin Microbiol 35:1948–1951, 1997. ble role in rabbit respiratory tract infections. Infect Immun
10. Zaoutis TE, Reinhard GR, Cioffe CJ, et al: Screening rabbit 35:1103–1109, 1982.
colonies for antibodies to Pasteurella multocida by an ELISA. 16. Knight DP, Paine JE, Speller DCE: A selective medium for
Lab Anim Sci 41:419–422, 1991. Pasteurella multocida and its use with animal and human
11. Klassen JM, Bernard BL, DiGiacomo RF: Enzyme-linked specimens. J Clin Pathol 36:591–594,1983.
immunosorbent assay for immunoglobulin G antibody to 17. Avril J, Donnio P, Pouedras P: Selective medium for Pasteurel-
Pasteurella multocida in rabbits. J Clin Microbiol 21:617– la multocida and its use to detect oropharyngeal carriage in
621, 1985. pig breeders. J Clin Microbiol 28:1438–1440, 1990.
12. Lukas VS, Ringler DH, Chrisp CE, Rush HG: An enzyme-
linked immunosorbent assay to detect serum IgG to Pas-
teurella multocida in naturally and experimentally infected
rabbits. Lab Anim Sci 37:60–64, 1987. About the Authors
13. Deeb BJ, DiGiacomo RF, Bernard BL, et al: Pasturella mul- Dr. Sanchez is affiliated with the Athens Diagnostic Labo-
tocida and Bordetella bronchiseptica infections in rabbits. J ratory, Drs. Mizan and Lee with the Department of Medi-
Clin Microbiol 28:70–75, 1990. cal Microbiology and Parasitology, and Dr. Ritchie with
14. Sanchez S, Mizan S, Quist CF, et al: Comparison of a new
serology test with PCR and culture for diagnosis of pasteurel- the Department of Small Animal Medicine and Surgery,
losis in rabbits. J Clin Microbiol, Submitted for publication, College of Veterinary Medicine, The University of Geor-
2000. gia, Athens, Georgia.
15. Glorioso JC, Jones GW, Rush HG, et al: Adhesion of type A