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Indirect hemagglutination test that uses glutaraldehyde-fixed sheep erythrocytes sensitized with extract antigens for detection of Pasteurella antibody.

Glutaraldehyde-fixed sheep erythrocytes (GA-SRBC) were used in the indirect hemagglutination test for the detection of Pasteurella antibody. GA-SRBC were stable for at least 6 months. Heat extract or potassium thiocyanate extract antigens of Pasteurella strains could be adsorbed onto GA-SRBC or tanned GA-SRBC, respectively. The indirect hemagglutination test reaction was capsular group specific with heat extract antigen-sensitized GA-SRBC but not potassium thiocyanate extract antigen-sensitized tanned GA-SRBC.

Animals↗

Molecular identification of Pasteurella dagmatis peritonitis in a patient undergoing peritoneal dialysis.

Pasteurella dagmatis was identified as the etiologic agent of peritonitis in a continuous ambulatory peritoneal dialysis patient by utilizing a molecular kit in our hospital's clinical laboratory. This method would appear a useful approach to identify a species of Pasteurella not included in the existing database of commercial identification kits when discrepancies exist between phenotypic tests.

Adult↗

Long-term respiratory tract infection with canine-associated Pasteurella dagmatis and Neisseria canis in a patient with chronic bronchiectasis.

Pasteurella dagmatis and Neisseria canis were repeatedly isolated from the sputum of a poodle-owning patient with chronic bronchiectasis. Commercially available systems failed to identify these unusual organisms: identification was made by 16S rRNA gene sequencing. Difficulties identifying these and five other canine-associated isolates (P. dagmatis [n = 2], Pasteurella canis [n = 2], and N. canis [n = 1]) are discussed.

Aged↗

The efficacy of sulbactam-ampicillin in the therapy of respiratory disease associated with ampicillin resistant Pasteurella species in housed calves.

Sulbactam-ampicillin is a combination of sulbactam, a beta-lactamase inhibitor, and ampicillin, a broad spectrum beta-lactam antibiotic. The efficacy of sulbactam-ampicillin was evaluated in the treatment of calf respiratory disease associated with ampicillin-sensitive and ampicillin-resistant strains of Pasteurella haemolytica and Pasteurella multocida. Treatment with sulbactam-ampicillin was compared with treatment with ampicillin alone in 123 Friesian calves, between three and five weeks old, exhibiting clinical signs of respiratory disease. Seven of the 59 calves treated with ampicillin died whereas only one death occurred in the 64 calves treated with sulbactam-ampicillin. In the calves which survived, treatment with sulbactam-ampicillin resulted in a significantly better clinical response, as measured by the reduction in severity of clinical signs. The results of bacteriological examinations indicated that there was a marked increase in the proportion of ampicillin-resistant isolates of P haemolytica subsequent to treatment with ampicillin, whereas the proportion of ampicillin-resistant isolates of P. haemolytica recovered from calves treated with sulbactam-ampicillin had declined. The superior efficacy of sulbactam-ampicillin observed in this study is explained by the inhibitory effect of sulbactam on beta-lactamases produced by resistant bacteria, thus rendering them susceptible to the ampicillin.

Ampicillin↗

Actinobacillus and Pasteurella species isolated from horses with lower airway disease.

Seventy-three bacterial isolates from 65 horses with and without evidence of lower airway disease were identified to assess whether the association with disease was accounted for by a small or large number of species. Just over half (50.5 per cent were Actinobacillus equuli, 17.8 per cent were A suis-like, 11 per cent were Pasteurella pneumotropica, 8.2 per cent were A lignieresii, 6.8 per cent were P haemolytica and 5.5 per cent were P mairii. These results suggest that a range of Actinobacillus and Pasteurella species can be isolated from the lower airways of horses, with many of the isolates being A equuli. Among the horses examined, lower airway inflammation was significantly associated with A suis-like bacteria and A lignieresii. However, these two species constituted too small a proportion of the undifferentiated Actinobacillus/Pasteurella species to explain the association of this group of bacteria with lower airway disease. Since A equuli constituted just over 50 per cent of the group of bacteria it is possible that it too is playing a significant role in lower airway disease.

Actinobacillus↗

Antigenic analysis of Pasteurella multocida (serotype 1) by crossed immunoelectrophoresis: characterization of cytoplasmic and cell envelope associated antigens.

The application of crossed immunoelectrophoresis to the analysis of a reference cytoplasm and cell envelope preparation from Pasteurella multocida serotype 1 revealed antigenic complexity not previously found. At least 55 cytoplasmic and 19 cell envelope antigens were clearly distinguished. Variation of anticytoplasm immunoglobulin concentration was a major determining factor in resolving the maximum array of cytoplasmic antigens. The use of intermediate gel modification of crossed immunoelectrophoresis permitted the recognition of antibodies in the preimmune rabbit serum against a number of cytoplasmic antigens and a single cell envelope antigen. This technique also demonstrated that reference cytoplasm obtained by 105,000 x g centrifugation of sonically disrupted pasteurellae and repeatedly washed reference cell envelope preparation contained antigens of either origin in amounts sufficient to elicit antibody responses in the host. Antisera to whole cells in the intermediate gel indicated that formalin killed P. multocida were capable of eliciting immune responses to both reference systems.

Animals↗

A protein toxin from Pasteurella multocida type D causes acute and chronic hepatic toxicity in rats.

Pasteurella toxin given subcutaneously to rats caused severe liver damage and growth suppression in doses as low as 15.6 ng. Toxin was lethal at and above 31.25 ng. Survival times were dose-dependent, and lesions differed with time of survival after toxin. Rats dead of acute toxicity had focal hepatic necrosis. Liver lesions were associated with diffuse endothelial damage, intravascular trapping of leukocytes, and degeneration of hepatocytes (characterized by glycogen depletion, development of vacuoles, and eosinophilic cytoplasmic inclusions). Endothelial cells, Kupffer cells, and macrophages had evidence of activation, e.g., increased cellular size with increases in Golgi vesicles, granules, and lysosomes. Rats with chronic toxicity (survival greater than 150 hr) had cirrhosis, intestinal villous atrophy, and markedly reduced body weight and fat. These data show that the rat is highly sensitive to toxins of Pasteurella multocida, and that even low doses of toxin cause liver injury and growth suppression.

Animals↗

Recovery of members of the Pasteurella-Actinobacillus-group from guinea pigs.

In the course of post-mortem bacteriological examination of conventional guinea pigs, 88 isolates belonging to the Pasteurella-Actinobacillus-group were recovered from 69 of 279 animals (25%). Most isolates were recovered from pneumonic lung, enteritic jejunum and inflamed mammary gland. No relationship was found between biotype and source of isolation. About 50% of isolates were recovered in pure culture or as the predominant micro-organism. It is concluded that members of the Pasteurella-Actinobacillus-group must be considered potentially pathogenic for guinea pigs.

Actinobacillus↗

Embryonic death in pregnant rats owing to intercurrent infection with Sendai virus and Pasteurella pneumotropica.

During an outbreak of Sendai virus infection in a colony of rats used for embryo production, severe lung lesions due to secondary colonization of the rat lungs with Pasteurella pneumotropica were noted. The effect on pregnancy was to cause embryonic death and resorption, leaving a deciduoma with trophoblastic giant cells as the only embryonic remnants. Up to 30% of fetuses in each pregnant animal were affected at the time of severe maternal lung lesions. Heavy growths of Pasteurella pneumotropica were obtained from the lungs of all affected dams. The formation of deciduomas as a result of embryonic death was due to the indirect effect of damage to the lungs during pregnancy rather than the direct pathogenic effect on the developing embryo of the microbial organisms isolated.

Animals↗

Elimination of Pasteurella pneumotropica from a contaminated mouse colony by oral administration of Enrofloxacin.

Enrofloxacin, a fluoroquinolone bactericidal antibiotic, was administered in an attempt to eradicate Pasteurella pneumotropica (P. pneumotropica) from a contaminated mouse colony. Contaminated mice, maintained within 4 animal rooms, were administered Enrofloxacin in drinking water at a daily dosage of 25.5 mg/kg for 2 weeks. Following one week of Enrofloxacin treatment, mice were selected randomly from each room and examined for P. pneumotropica. This procedure was repeated two or three times until all mice examined tested negative for the Pasteurella strain. With the exception of one room, treated mice consistently tested negative for P. pneumotropica for up to 45 weeks following completion of Enrofloxacin treatment. Thus, oral administration of Enrofloxacin significantly eliminated P. pneumotropica from a contaminated mouse colony.

Administration, Oral↗

Pasteurella multocida bacteremia: report of thirteen cases over twelve years and review of the literature.

13 episodes of bacteremia caused by Pasteurella multocida were seen in a general hospital during a 12-year period. All the patients had an underlying disease (77% had cirrhosis) and 2 were receiving chemotherapy for hematologic malignancy. There was a numerical preponderance of male patients (69%). In 5/13 cases a recent animal-derived trauma could be found. In the other cases the source of the infecting organism was thought to be endogenous (from patients' own pharyngeal commensal flora) or secondary to contact with secretions of a pet animal. The clinical presentation of sepsis caused by this organism was nonspecific. Hypotension was seen in 5 cases. Localized sites of infection were certain in 6 and only clinically suspected in 4 other cases. The overall mortality rate was 31%. The administration of ampicillin seems the appropriate therapy for Pasteurella multocida bacteremia.

Anti-Bacterial Agents↗

Pasteurella multocida: a rare case of shunt infection.

Pasteurella multocida, the major pathogen following an animal bite, is a rare cause of intracranial infection. This report documents the first case of ventriculo-peritoneal shunt infection with Pasteurella multocida.

Adult↗

Experimental contact transmission of Pasteurella haemolytica from clinically normal domestic sheep causing pneumonia in Rocky Mountain bighorn sheep.

Two Rocky Mountain bighorn lambs (Ovis canadensis canadensis) were held in captivity for 120 days before being housed with two domestic sheep. The lambs were clinically normal and had no Pasteurella spp. on nasal swab cultures. The domestic sheep were known to carry Pasteurella haemolytica biotype A in the nasal passages. After being in close contact for 19 days. P. haemolytica biotype A was cultured from nasal swabs of one of the bighorn lambs. By 26 days, both bighorn sheep developed coughs, were anorectic and became lethargic and nasal swabs yielded P. haemolytica biotype T, serotype 10. Twenty-nine days after contact, the lambs were necropsied and found to have extensive fibrinous bronchopneumonia. From affected tissues pure cultures of beta-hemolytic P. haemolytica biotype T, serotype 10 were grown. Both domestic sheep remained clinically normal and had no gross or microscopic lesions, but they carried the same P. haemolytica serotype in their tonsils. Behavioural observations gave no indication of stress in the bighorn lambs.

Animals↗

A cryopreservation method for Pasteurella multocida from wetland samples.

A cryopreservation method and improved isolation techniques for detection of Pasteurella multocida from wetland samples were developed. Wetland water samples were collected in the field, diluted in dimethyl sulfoxide (DMSO, final concentration 10%), and frozen at -180 C in a liquid nitrogen vapor shipper. Frozen samples were transported to the laboratory where they were subsequently thawed and processed in Pasteurella multocida selective broth (PMSB) to isolate P. multocida. This method allowed for consistent isolation of 2 to 18 organisms/ml from water seeded with known concentrations of P. multocida. The method compared favorably with the standard mouse inoculation method and allowed for preservation of the samples until they could be processed in the laboratory.

Animals↗

Immunologic responses of domestic and bighorn sheep to a multivalent Pasteurella haemolytica vaccine.

The efficacy of a Pasteurella haemolytica vaccine (serotypes A1, A2, and T10) to induce humoral antibodies and alter colonization of the upper respiratory tract by related P. haemolytica spp. strains was evaluated in 10 bighorn (Ovis canadensis canadensis) and 10 domestic (Ovis aries) sheep. Sheep of each species were divided into five pairs based on age and history of respiratory disease. One sheep in each pair was vaccinated twice 2 wk apart with 2 ml of vaccine (VAC group) and the remaining animals (NV group) were injected with 2 ml of sterile saline. Mild, transient lameness was the only observed adverse effect. Blood sera from the sheep were tested for agglutinating antibodies against whole cells of A1, A2, and T10 and for leukotoxin neutralizing antibodies. Antibody titers were expressed as the reciprocal log2 of the highest reactive dilutions. Domestic sheep > 1-yr-old and two bighorn sheep with a history of A1 infection had higher titers throughout the study against A1 cells than domestic sheep < 1-yr-old and bighorns without a history of A1 infection. Both domestic and bighorn sheep had log2 titers of 8 to 12 against A2 cells and 6 to 12 against T10 cells during this time. Bighorn sheep in the VAC group had 2 to 32 fold titer increases for A1 cells by 2 wk post-vaccination (PV) compared to 0 to 2 fold increases in VAC domestic sheep. Two to 16 and 0 to 8 fold increases in antibodies titers to A2 and T10 cells, respectively, were detected in sera of both VAC groups. Sera of bighorn sheep with a history of respiratory disease and all domestic sheep had log2 leukotoxin neutralizing antibody titers of 4 to 14 in contrast to < or = 2 in sera of bighorn sheep without a history of respiratory disease. Neutralizing antibody titers of two bighorns without a history of respiratory disease in the VAC group increased from log2 0 to 5 in one and from 0 to 9 in the other 2 wk PV. Antibody increases in these animals were no longer evident at 16 wk PV while titers of animals with histories of disease remained relatively stable. The types and numbers of Pasteurella spp. isolated from nasal and pharyngeal swabs varied throughout the study without conclusive evidence of suppression of colonization. Although the animals were not experimentally challenged to determine the efficacy of the vaccine, one VAC and one NV bighorn sheep died following introduction of an A2 P. haemolytica strain when leukotoxin neutralizing antibodies had returned to pre-vaccination levels. This vaccine appeared to be safe for use in bighorn sheep and stimulated moderate but transient increases in antibody levels which should provide some protection against naturally occurring disease. A vaccine which would induce production of high and maintained antibodies against multiple strains of P. haemolytica would be valuable for use in bighorn sheep maintained in captivity or when captured for relocation.

Agglutination Tests↗

Detection of annexin I and IV and haptoglobin in bronchoalveolar lavage fluid from calves experimentally inoculated with Pasteurella haemolytica.

OBJECTIVE: To determine whether annexins or haptoglobin could be detected in bronchoalveolar lavage (BAL) fluid specimens obtained from calves experimentally inoculated with Pasteurella haemolytica. ANIMALS: Twelve 2- to 3-month-old male Holstein calves. PROCEDURE: Pasteurella haemolytica was inoculated into the right lung lobes of each of 6 calves. Six other calves received vehicle alone and were used as control calves. Specimens of BAL fluid were obtained from 3 control and 3 inoculated calves 1 day after inoculation and from the other calves 2 days after inoculation. The amount of annexins I, II, IV, and VI, and haptoglobin in BAL fluid specimens was examined by use of immunoblot analysis. RESULTS: Annexins I and IV were detected in BAL fluid specimens obtained from the right lung lobes of each of the inoculated calves, but annexins II and VI were not. Annexin I also was found in BAL fluid specimens obtained from the left lung lobes of each inoculated calf and from left and right lung lobes of the control calves. By comparison, detection of annexin IV was essentially limited to the right lung lobes of inoculated calves. Haptoglobin was detected in some, but not all, BAL fluid specimens from the right lung lobes of inoculated calves, and its detection in BAL fluid was associated with serum proteins such as albumin. CONCLUSIONS AND CLINICAL RELEVANCE: Annexin IV was detected most specifically in response to inoculation of P haemolytica. This protein could be used as a marker for inflammatory pulmonary disease caused by P haemolytica.

Animals↗

Experimentally induced pneumonia in scid/beige mice, using a bovine isolate of Pasteurella haemolytica.

BACKGROUND AND PURPOSE: Intranasal challenge of immunocompetent mice with Pasteurella haemolytica results in little or no pulmonary inflammation. The study reported here was designed to investigate the inflammatory response in the lungs of immunodeficient scid/beige mice after similar challenge. METHODS: Fifty-five scid/beige mice were challenged intranasally with saline or one of three doses (2.8 x 10(6), 3.4 x 10(9), or 3.3 x 10(11) colony-forming units [CFU]/ml) of P. haemolytica. The lungs were examined for changes in weight, bacterial count, and presence of gross and microscopic lesions at 24, 48, or 96 hours after challenge. RESULTS: Intranasal challenge with concentrations > or = 3.4 x 10(9) CFU/ml of P. haemolytica induced significantly heavier lung weight, with severe pulmonary lesions, and development of suppurative and fibrinous bronchopneumonia in dose- and time-dependent manner 48 hours after challenge. Pasteurella haemolytica was consistently isolated from the lungs at 24 hours after challenge. CONCLUSIONS: Bronchopneumonia was induced by P. haemolytica in mice without manipulation of the mouse or the bacteria. The lesions were similar to those that develop in the lungs of cattle infected with P. haemolytica and indicate potential use of the model for the study of this host/bacterial interaction.

Animals↗