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Detecting nonhemolytic Pasteurella haemolytica infections in healthy Rocky Mountain bighorn sheep (Ovis canadensis canadensis): influences of sample site and handling.

Effects of sampling procedures on ability to culture Pasteurella spp. from Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were examined experimentally. Sample site influenced (P less than 0.0001) recovery of P. haemolytica in adult bighorn sheep. We isolated nonhemolytic P. haemolytica from 18 of 19 tonsillar swabs and 18 of 19 tonsillar biopsies from adult sheep, yet only four of 19 nasal swabs yielded isolates. Sample handling also affected (P less than 0.0001) recovery of P. haemolytica. Nonhemolytic P. haemolytica was cultured from 14 of 19 tonsillar swabs plated directly onto blood agar, but from only two of 19 swabs stored for 24 hr in modified Stuart's medium. We detected nonhemolytic P. haemolytica at least once in bronchial aspirates from four and in nasal swabs from three of six bighorn lambs. Based on direct cultures of tonsillar swabs and/or biopsies, all 26 bighorn sheep (seven lambs, 19 adults) sampled were infected with nonhemolytic P. haemolytica; only two lambs developed pneumonia during the study period. Thirty-four of 37 nonhemolytic P. haemolytica isolates tested were biotype T; three were biotype A. Serotypes 3; 4; 3, 4 and 3, 4, 10 were identified in a subsample of 17 isolates. Our data suggest tonsillar swabs or biopsies plated directly onto blood agar and incubated immediately offer the greatest probability of recovering nonhemolytic P. haemolytica from health bighorn sheep.

Animals↗

Molecular characterization of Pasteurella testudinis isolated from desert tortoises (Gopherus agassizii) with and without upper respiratory tract disease.

Isolates of Pasteurella testudinis recovered from clinically healthy desert tortoises (Gopherus agassizii) and tortoises with upper respiratory tract disease (URTD) were characterized in an attempt to identify strains associated with disease. Eighty-nine isolates, 52 from ill and 37 from healthy tortoises collected from Nevada (USA), June 1990 to September 1991, were genomically fingerprinted and grouped based on ribotype similarity. Twelve isolates (six from ill and six from healthy tortoises) were further characterized with regard to whole-cell protein (WCP) and outer membrane protein (OMP) composition and their ability to survive in normal tortoise plasma. The 89 isolates were initially distributed into 33 distinct ribotype groups using the restriction enzyme EcoRI; five ribotypes contained over 50% of the isolates. Only one EcoRI ribotype was comprised of multiple isolates (n = 4) exclusively recovered from tortoises with URTD. When the ten EcoRI ribotypes that contained more than one isolate per ribotype were further studied using a second restriction enzyme, EcoRV, one EcoRI/EcoRV ribotype contained five isolates recovered from URTD tortoises and none from healthy animals. The EcoRI ribotype comprised of four isolates, all from tortoises with URTD, was further separated into three distinct groups with EcoRV. All 12 isolates studied grew equally well in normal tortoise plasma, and when broth-grown WCP and OMP profiles were evaluated, no proteins were unique to isolates from URTD tortoises. Iron-regulated OMP's were produced in three isolates examined, but these OMP's apparently were not virulence-related.

Animals↗

Pasteurella multocida serotype 1 isolated from a lesser snow goose: evidence of a carrier state.

Pharyngeal swabs were collected from 298 lesser snow geese (Chen caerulescens caerulescens) at Banks Island (Northwest Territories. Canada) in the summer of 1994. Pasteurella multocida serotype 1 was isolated from an adult male bird and P. multocida serotype 3 was isolated from an adult female goose. Pathogenicity of the serotype 1 isolate was confirmed by inoculation in Pekin ducks (Anas platyrhynchos). The serotype 3 isolate was non-pathogenic in Pekin ducks. This is the first documented isolation of pathogenic P. multocida serotype 1 from apparently healthy wild snow geese.

Animals↗

Antibodies against Pasteurella multocida in snow geese in the western Arctic.

To determine if lesser snow geese (Chen caerulescens caerulescens) are a potential reservoir for the Pasteurella multocida bacterium that causes avian cholera, serum samples and/or pharyngeal swabs were collected from > 3,400 adult geese breeding on Wrangel Island (Russia) and Banks Island (Canada) during 1993-1996. Pharyngeal swab sampling rarely (> 0.1%) detected birds that were exposed to P. multocida in these populations. Geese with serum antibody levels indicating recent infection with P. multocida were found at both breeding colonies. Prevalence of seropositive birds was 3.5% at Wrangel Island, an area that has no recorded history of avian cholera epizootics. Prevalence of seropositive birds was 2.8% at Banks Island in 1994, but increased to 8.2% during 1995 and 1996 when an estimated 40,000-60,000 snow geese were infected. Approximately 50% of the infected birds died during the epizootic and a portion of the surviving birds may have become carriers of the disease. This pattern of prevalence indicated that enzootic levels of infection with P. multocida occurred at both breeding colonies. When no avian cholera epizootics occurred (Wrangel Island, Banks Island in 1994), female snow geese (4.7%) had higher antibody prevalence than males (2.0%).

Animals↗

Comparison of methods to detect Pasteurella multocida in carrier waterfowl.

We conducted laboratory challenge trials using mallard ducks (Anas platyrhynchos) to compare methods for detecting carriers of Pasteurella multocida, the bacterium that causes avian cholera, in wild birds. Birds that survived the initial infection were euthanized at 2-4 wk intervals up to 14 wk post challenge. Isolates of P. multocida were obtained at necropsy from 23% of the birds that survived initial infection. We found that swab samples (oral, cloacal, nasal, eye, and leg joint) were most effective for detecting carrier birds up to 14 wk post infection. No detectable differences in isolation were observed for samples stored in either 10% dimethysulfoxide or brain heart infusion broth. The frequency of detecting carriers in our challenge trials appeared to be related to mortality rates observed during the trial, but was not related to a number of other factors including time after challenge, time delays in collecting tissues postmortem, and route of infection. In our trials, there was little association between antibody levels and carrier status. We concluded that swabs samples collected from recently dead birds, stored in liquid nitrogen, and processed using selective broth provide a feasible field method for detecting P. multocida carriers in wild waterfowl.

Animals↗

Characterization of Pasteurella multocida associated with pneumonia in bighorn sheep.

Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Although P. multocida is not ordinarily associated with disease in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), numerous isolates were cultured in high numbers from free-ranging bighorn sheep in the Hells Canyon area of Idaho, Washington, and Oregon (USA) during the winter of 1995-96. Animals captured in Hells Canyon and held in captivity, and their offspring, also harbored P. multocida. Biochemical utilization tests on 90 isolates identified three subspecies: P. multocida multocida a (n = 54); P. multocida multocida b (n = 13); and P. multocida gallicida (n = 15); and a non-speciated biotype, U6 (n = 8). Genomic DNA digestion with restriction endonuclease Hha I separated the isolates into 62 unique restriction fragment length polymorphism profiles. Capsular type A was predominant (72% of isolates). Only one isolate type, which may have been transmitted from a feral goat, was capsular type D, possessed the structural gene, toxA, for dermonecrotoxin detected by polymerase chain reaction, and produced toxin as determined by monoclonal antibody immunoblot. In conclusion, bighorn sheep in this study carried diverse types of generally non-toxigenic P. multocida associated with epizootic pneumonia.

Animals↗

Characterization of Pasteurella multocida isolates from wetland ecosystems during 1996 to 1999.

We cultured 126 Pasteurella multocida isolates, 92 from water and 34 from sediment samples collected from wetlands in the Pacific and Central flyways of the United States between 1996 and 1999. Most (121) of the isolates were P. multocida serotype 1, but serotypes 3, 3/4, 10, and 11 were also found. Many (82) of the isolates were further characterized by DNA fingerprinting procedures and tested in Pekin ducks for virulence. Almost all the serotype 1 isolates we tested caused mortality in Pekin ducks. Serotype 1 isolates varied in virulence, but the most consistent pattern was higher mortality in male ducks than in females. We found no evidence that isolates found in sediment vs. water, between Pacific and Central flyways, or during El Niño years had consistently different virulence. We also found a number of non-serotype 1 isolates that were avirulent in Pekin ducks. Isolates had DNA fingerprint profiles similar to those found in birds that died during avian cholera outbreaks.

Animals↗

Pasteurella multocida from outbreaks of avian cholera in wild and captive birds in Denmark.

An outbreak of avian cholera was observed among wild birds in a few localities in Denmark in 2001. The highest mortalities were among breeding eiders (Somateria mollissima) and gulls (Larus spp.). Pulsed-field gel electrophoresis (PFGE) was conducted using ApaI and SmaI as restriction enzymes and restriction enzyme analysis (REA) using HpaII. The Pasteurella multocida subsp. multocida strain isolated from birds in this outbreak was indistinguishable from a strain that caused outbreaks in 1996 and 2003. Most isolates from domestic poultry had other PFGE patterns but some were indistinguishable from the outbreak strain. Among 68 isolates from wild birds, only one PFGE and one REA pattern were demonstrated, whereas among 23 isolates from domestic poultry, 14 different SmaI, 12 different ApaI, and 10 different HpaII patterns were found. The results suggest that a P. multocida strain has survived during several years among wild birds in Denmark.

Animals↗

Associations between water quality, Pasteurella multocida, and avian cholera at Sacramento National Wildlife Refuge.

We studied patterns in avian cholera mortality, the presence of Pasteurella multocida in the water or sediment, and water chemistry characteristics in 10 wetlands at the Sacramento National Wildlife Refuge Complex (California, USA), an area of recurrent avian cholera epizootics, during the winters of 1997 and 1998. Avian cholera outbreaks (>or=50 dead birds) occurred on two wetlands during the winter of 1997, but no P. multocida were recovered from 390 water and 390 sediment samples from any of the 10 wetlands. No mortality events were observed on study wetlands during the winter of 1998; however, P. multocida was recovered from water and sediment samples in six of the 10 study wetlands. The pH levels were higher for wetlands experiencing outbreaks during the winter of 1997 than for nonoutbreak wetlands, and aluminum concentrations were higher in wetlands from which P. multocida were recovered during the winter of 1998. Water chemistry parameters (calcium, magnesium, sodium, and dissolved protein) previously linked with P. multocida and avian cholera mortality were not associated with the occurrence of avian cholera outbreaks or the presence of P. multocida in our study wetlands. Overall, we found no evidence to support the hypothesis that wetland characteristics facilitate the presence of P. multocida and, thereby, allow some wetlands to serve as long-term sources (reservoirs) for P. multocida.

Animals↗

Persistence of Pasteurella multocida in wetlands following avian cholera outbreaks.

Avian cholera, caused by Pasteurella multocida, affects waterbirds across North America and occurs worldwide among various avian species. Once an epizootic begins, contamination of the wetland environment likely facilitates the transmission of P. multocida to susceptible birds. To evaluate the ability of P. multocida serotype-1, the most common serotype associated with avian cholera in waterfowl in western and central North America, to persist in wetlands and to identify environmental factors associated with its persistence, we collected water and sediment samples from 23 wetlands during winters and springs of 1996-99. These samples were collected during avian cholera outbreaks and for up to 13 wk following initial sampling. We recovered P. multocida from six wetlands that were sampled following the initial outbreaks, but no P. multocida was isolated later than 7 wk after the initial outbreak sampling. We found no significant relationship between the probability of recovery of P. multocida during resampling and the abundance of the bacterium recovered during initial sampling, the substrate from which isolates were collected, isolate virulence, or water quality conditions previously suggested to be related to the abundance or survival of P. multocida. Our results indicate that wetlands are unlikely to serve as a long-term reservoir for P. multocida because the bacterium does not persist in wetlands for long time periods following avian cholera outbreaks.

Animals↗

Pasteurella anatipestifer as a cause of mortality in captive wild waterfowl.

An outbreak of Pasteurella anatipestifer infection in young wild waterfowl at the Niska Waterfowl Research Center resulted in losses of approximately 100 Blue and Snow Geese, one White-fronted Goose, five Mandarin Ducks, one Black Duck and one Wood Duck. Clinical signs included diarrhea, paralysis and tremors. Gross lesions were fibrin deposits on serosal surfaces, hemorrhages on the epicardium, consolidation of the lungs, cloudy or flaky deposits on the air sacs, and dark, swollen spleens. Microscopic lesions included fibrinous meningitis, pneumonitis, air saculitis and serositis. Swollen leg and foot joints were seen in some cases. Chloramphenicol treatment seemed to be of benefit in controlling the outbreak.

Animals↗

[Murine model for experimental studies on attenuated mutants of a Pasteurella multocida strain with pathogenicity for the calf. 2. Primary immunization against lethal infection, using fully attenuated streptomycin-dependent (Sm-d) mutant and booster immunization with partially attenuated streptomycin-dependent (Sm-id) revertants].

An experimental study was conducted into the immunisation of mice to a strain of Pasteurella multocida with pathogenicity for calf. Here are the findings: (a) The attenuation of Sm-id revertants was measured with reference to the order of extinction and compared to the wild strain. (b) A selected range of Sm-id revertants with differentiated attenuation is presented. (c) Single immunisation, using fully attenuatted Sm-d mutant does prevent lethal infection with partially attenuated Sm-id revertants, although it is ineffective against wild strain infection. (d) Immunity against 100 times the lethal dose of the wild strain can be achieved by two immunisations, that is primary antigen application, using fully attenuated Sm-d mutant, and booster immunisation, using partially attenuated Sm-id revertant.

Animals↗

Safety and efficacy of two live Pasteurella multocida aro-A mutant vaccines in chickens.

Two auxotrophic aro-A mutants of Pasteurella multocida designated PMP1 (serotype 1) and PMP3 (serotype 3) were tested as vaccine candidates to protect chickens against fowl cholera. A reliable intratracheal challenge method was established that resulted in > or = 75% mortality in both specific-pathogen-free chickens and commercial broiler breeders 24 hr after challenge. Dose protection studies indicated that at least 10(6) colony-forming units (CFU) of PMP1 and 10(8) CFU of PMP3 were required to provide complete protection against challenge in all birds. Although high doses of 10(9) CFU of the vaccine strains produced some endotoxinlike reactions, lower but protective dose levels produced no clinical sign or lesion in any chicken. Both vaccine strains provided cross-protection with a heterologous challenge strain PM206 (serotype 4). Future studies will examine the duration of protective immunity induced by the two vaccine candidates, PMP1 and PMP3, and cross-protection against other serovars.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗

The effect of route of inoculation on the virulence of raptorial Pasteurella multocida isolates in Pekin ducks (Anas platyrhynchos).

The purpose of this study was to determine the virulence of raptorial Pasteurella multocida for ducks and the effect of various routes of inoculation on virulence. Four-week-old Pekin ducks (Anas platyrhynchos) were challenged with one of three raptorial isolates (RTHA-2, RTHA-4, or WESO-1) by one of five inoculation routes (intranasal, intraocular, intravenous, oral, and subcutaneous). Ducks were monitored daily for mortality until 2 wk postchallenge. Results indicated that the intravenous route caused the most mortality for all isolates and that significant variation existed in the virulence among the sources of P. multocida, with WESO-1 causing the least mortality of the isolates tested.

Animals↗

[Murine model for experimental studies on attenuated mutants of a Pasteurella multocida strain with pathogenicity for the calf. 1. Isolation and biological characterization of streptomycin-dependent mutants (Sm-d) and their examination for immunogenicity in comparison with thermally inactivated pathogens].

An experimental study was conducted into the immunisation of mice to a strain of Pasteurella multocida with pathogenicity for calf. Here are the results: (a) Stable Sm-d mutants were isolated. (b) The genetic stability of this mutant type was established by unsubstantiable reversion up to the magnitude of 10(-8) or - in cases in which 20 Sm-id revertants maximum occurred in 10(-8) Sm-d germs--by prolongation of the generation period of these suppressor mutants which can be defined also as minus mutants. (c) Stable attenuation of selected Sm-d mutants has been established in mice by intraperitoneal application of 108 germs. (d) Live germs with restricted reproduction exhibited more immunogenicity to thermally inactivated antigen.

Animals↗

Evaluation of commercially available Escherichia coli J5 bacterin as protection against experimental challenge with Pasteurella multocida in rabbits.

OBJECTIVE: To evaluate the ability of commercially available Escherichia coli J5 bacterin to protect rabbits from experimental challenge with Pasteurella multocida. ANIMALS: 40 P multocida-free New Zealand White rabbits. PROCEDURES: Rabbits were assigned to 1 of 4 groups of 10 rabbits each. Three of the groups were inoculated SC with J5 bacterin at 8 weeks old. Inoculation was repeated 3 and 6 weeks later. The fourth group was not inoculated and served as controls. Groups 1, 2, and 3 were given 10(9), 10(8), and 10(7) colony forming units (CFU), respectively. Response was monitored by titer assessment, using an E coli J5 antigen capture ELISA. Five weeks after the last inoculation, all rabbits were challenged with P multocida and observed for an additional 5 weeks. Clinical, hematologic, serologic, culture, and necropsy data were collected. RESULTS: Inoculation of rabbits with 10(9) CFU of E coli J5 bacterin-induced titers that were significantly greater than titers of rabbits vaccinated with 10(8) or 10(7) CFU or those in controls. The incidence of acute bacteremia was lower in rabbits with high titers. At necropsy, prevalence of lesions typical of P multocida was not significantly different among groups. Prevalence of histologic lesions was also not significantly different among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Although the bacterin induced considerable antibody response and possibly reduced the rate of bacteremia, antibodies were not protective against long-term colonization or infection of the frontal sinuses or tympanic bullae by the challenge strain of P multocida. This bacterin in its currently available form is unlikely to aid in reducing the prevalence of pasteurellosis in rabbits.

Animals↗

Effect of aerial ammonia on porcine infection of the respiratory tract with toxigenic Pasteurella multocida.

The objective of the experimental study was to examine whether aerial ammonia alone could predispose the respiratory system of pigs to infection with toxigenic Pasteurella multocida type A. Two groups of 5 pigs each were continuously exposed to 50 ppm ammonia and less than 5 ppm ammonia, respectively, for a 59-day period (from 37 kg to 90 kg bodyweight) followed by necropsy. In an aerosol chamber all pigs were exposed to an aerosol of toxigenic P. multocida type A (mean bacterial concentration in the aerosol-exposure chamber: 10(5) colony forming units/m3; exposure period: 25 min) at day 10, 21, 35 and 49 after the onset of ammonia exposure. During the experiment none of the pigs showed clinical signs of pneumonia nor did they develop visible distortion of the snout. None of the pigs had gross lesions in the lungs at necropsy and toxigenic P. multocida was not detected by culture from the lungs from any of the pigs. The chance of recovering toxigenic P. multocida from nasal swabs (collected during experiment) was 2-4 times greater in the test group compared to the control group. The average daily weight gain was lower for the ammonia exposed pigs compared to the control group. In conclusion the results from this study suggest that ammonia in concentrations of 50 ppm is unlikely to predispose growing pigs to pulmonary infection with toxigenic P. multocida.

Aerosols↗

[Pasteurella spp: a new microorganism to the cause of acute suppurative thyroiditis].

Acute suppurative thyroiditis is an uncommon disease due to local resistance of the gland to infection. Preexisting gland pathology and local anatomic abnormalities are predisposing factors. We present the first case described in the medical literature caused by Pasteurella spp after upper respiratory infection with insidious manifestations resembling subacute thyroiditis. The course was benign after surgical drainage.

Acute Disease↗