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PubMed1980

Articular and skeletal infections caused by Pasteurella multocida.

Pasteurella multocida infections of joints and bones generally occur in individuals who have contact with cats or dogs. Osteomyelitis usually follows penetrating trauma such as an animal bite. Septic arthritis tends to occur in patients who have preexisting inflammatory joint disease, especially if a systemic condition which is known to predispose to infection is present. The principles of therapy for septic arthritis or osteomyelitis are no different from those which have been established for other infecting organisms. Although P multocida is susceptible in vitro to penicillin, treatment of septic arthritis with this drug is still associated with a slow therapeutic response.

Anti-Bacterial Agents↗
PubMed1992

Septic arthritis due to Pasteurella multocida.

We have reported two cases of septic arthritis caused by Pasteurella multocida, the first, septic polyarthritis unrelated to animal contact and the other, septic monarthritis after a cat scratch. Serious infections with this gram-negative bacillus must be treated with intravenous penicillin, ampicillin, or a third-generation cephalosporin; aminoglycosides are not effective. A high index of suspicion is appropriate when there is a history of a recent animal-inflicted wound, though infection may also occur with either nontraumatic animal contact or no recognized animal contact at all.

Adult↗
PubMed1992

Bacteremia due to Pasteurella multocida.

Pasteurella multocida should be considered as a possible etiologic agent in any infection that is the result of an animal bite or scratch. Because of its opportunistic capability, it should be included among the possible pathogens in bacteremia, particularly in any patient with immunosuppression or liver cirrhosis, especially if there is a history of animal exposure.

Adolescent↗
PubMed2004

Pasteurella multocida meningitis and cervical spine osteomyelitis in a neonate.

A 20-day-old male infant presented with fever, decreased alertness and quadriparesis as a result of Pasteurella multocida meningitis and C1-2 vertebral osteomyelitis. Although his household contained 2 pet cats, there was no history of bites, scratches or licks. We speculate that colonization of the nasopharynx was followed by contiguous spread to the retropharyngeal soft tissue, cervical vertebrae and meninges.

Anti-Bacterial Agents↗
PubMed2004

Spontaneous bacterial peritonitis with Pasteurella multocida in cirrhosis: case report and review of literature.

Most Pasteurella multocida human infections involve skin and soft tissues and invariably develop after a bite or a scratch from a dog or a cat. However, other infections with this organism occur infrequently. Enteric microorganisms are the common cause of spontaneous bacterial peritonitis (SBP). We report a case of SBP in a cirrhotic patient from P. multocida. English literature (Pubmed) review revealed 12 adult cases of SBP in cirrhotic patients with P multocida. Nine patients were exposed to animals, though a break in the skin or a bite was not reported in each case. The SBP was fatal in four of these patients.

Adult↗
PubMed1979

Immunogenicity of a chaotropically extracted protective antigen(s) of Pasteurella multocida type A (bovine origin) against experimental pasteurellosis in mice.

Immunization of mice with a potassium thiocyanate extract of Pasteurella multocida type A (bovine strain) imparted complete protection against an experimental challenge infection with 1.6 x 10(3) colony-forming units of the homologous strain. Formalin-treated vaccine (one or two doses) was much less effective. Significantly higher bactericidal antibody titres were observed in the sera of mice immunized with the extract compared with those given the vaccine. The bactericidal activity was localized in the IgM and IgG fractions. The extract consisted of protein(s) and carbohydrate(s) and on immunoelectrophoresis displayed at least three antigenetically distinct components.

Animals↗
PubMed1984

Comparison of cell surface antigen extracts from two serotypes of Pasteurella haemolytica.

Cells of Pasteurella haemolytica serotypes A1 and A6 were extracted with sodium salicylate and the chemical and antigenic composition of both extracts determined. The extracts were concentrated by ultrafiltration and the serotype antigen, measured by the indirect haemagglutination test, was estimated to have a molecular weight between 100 000 and 300 000. The chemical composition of sodium salicylate extracts (SSEs) from both serotypes was similar, having protein, carbohydrate, fatty acid and phosphorus present in the ratio 10:1:0.5:0.1. SDS-PAGE of both SSEs gave similar profiles with at least 48 bands present. These results suggest that sodium salicylate removes the outer membrane of P. haemolytica. Crossed immunoelectrophoresis indicated that a major serotype-specific antigen was present in SSEs of both strains. This antigen was extracted from the SSE with hot phenol/water and analysed by gas chromatography. The sugar composition of A1 and A6 phenol/water extract (PWE) was qualitatively identical although some differences in proportions were observed. A1 and A6 PWE antigens protected mice against homologous serotype challenge and A6 PWE protected against heterologous (A1) challenge.

Animals↗
PubMed1989

Iron-repressible outer-membrane proteins of Pasteurella haemolytica.

The outer-membrane protein (OMP) profile of Pasteurella haemolytica grown under iron-replete and iron-restricted conditions was studied by polyacrylamide gel electrophoresis and immunoblotting. A serotype 1 isolate induced the synthesis of a new 77,000 Mr OMP in iron-restricted media while two other proteins of 100,000 Mr and 71,000 Mr were synthesized in increased amounts. None of these proteins were peptidoglycan-associated or heat-modifiable, and only the 100,000 Mr protein showed some degree of disulphide cross-linking. Kinetic analysis revealed that the iron-repressible proteins appeared in the outer membrane within 15 min of establishment of iron-restricted conditions. Analysis of P. haemolytica isolates representing serotypes 1 to 12 showed that iron-repressible OMPs of 77,000 Mr and 71,000 Mr could be induced in all 12 serotypes but that there was some variability in the expression of the 100,000 Mr protein. Immunoblotting of OMPs with convalescent sera from P. haemolytica-infected calves indicated that antibodies directed against all three iron-repressible OMPs were present, suggesting that these proteins were expressed in vivo.

Animals↗
PubMed1992

Pathogenic activities of live cells and extracellular products of the fish pathogen Pasteurella piscicida.

The pathobiological activities in vivo and in vitro of live cells and extracellular products (ECP) of eleven Pasteurella piscicida strains of different origin were examined. Infectivity trials showed that P. piscicida did not possess strict host specificity since the majority of the isolates were virulent for gilthead seabream, rainbow trout and turbot, with LD50 values ranging between 10(3) and 10(6) live cells. However, none of the strains tested were pathogenic for mice (LD50 > 10(8) cells)). In addition, the ECP were strongly toxic for fish (LD50 ranging from 1.0 to 4.6 micrograms protein per g fish), which clearly demonstrates their important role in the pathogenesis of pasteurellosis. All the ECP samples were cytotoxic for fish and homoiothermic cell lines, possessed notable phospholipase activity and displayed haemolytic activity for sheep, salmon and turbot erythrocytes (but not for trout erythrocytes). However, the production of proteolytic enzymes differed among the P. piscicida strains. Although no strain displayed elastase activity, five isolates (the Japanese and Italian strains) hydrolysed casein and gelatin. All these biological activities in vivo and in vitro were lost after heat treatment (100 degrees C for 10 min). The general enzymic patterns of both live cells and ECP evaluated by the API-ZYM system also revealed some variation among the P. piscicida isolates. Generally, whole cells showed a wider range of enzymic activities than ECP. The results presented here are important for the selection of strains in the development of effective polyvalent pasteurellosis vaccines containing both whole cells and ECP.

Animals↗
PubMed1997

Genetic relationships among Pasteurella trehalosi isolates based on multilocus enzyme electrophoresis.

Genetic diversity among 60 British Pasteurella trehalosi isolates representing the four recognized capsular serotypes, T3, T4, T10 and T15, and recovered predominantly from sheep suffering from systemic pasteurellosis, was estimated by analysing electrophoretically demonstrable allelic variation at structural genes encoding 19 enzymes. Thirteen of the locl were polymorphic and 20 distinctive multilocus genotypes (electrophoretic types, ETs) were identified. The population structure of P. trehalosi is clonal and its genetic diversity is limited compared with most other pathogenic bacteria. ETs represent clones, and isolates of the same ET were generally associated with the same combination of serotype, LPS type and outer-membrane protein (OMP) type. The genetic diversity of isolates within each of the capsular serotypes varied. Serotype T10 was represented by 18 isolates in two related ETs and exhibited little diversity. By contrast, serotype T15 was represented by 18 isolates in nine ETs and was almost as diverse as the species as a whole Serotype T4 was represented by 18 isolates in five ETs and was less diverse than serotype T15. Although serotype T3 was more diverse than serotype T15 it was represented by only three isolates. With the exception of the T10 isolates and those recovered from healthy sheep, 35 disease isolates belonged to 16 ETs, each of which was represented by only one to four isolates. The fact that a high proportion of disease is caused by a relatively large number of clones suggests that P. trehalosi is essentially an opportunistic pathogen. In addition to having the same capsular structure, isolates belonging to the two T10 clones were characterized by possession of similar, if not identical, O-antigens (LPS types 2 and 4). The occurrence of 18 serotype T10 isolates in only two ETs suggests that the T10 capsule and type 2/4 O-antigen confer enhanced virulence on members of these two clones. Multilocus enzyme electrophoresis (MLEE) had greater resolving power than did capsule/LPS/OMP analysis, being able to distinguish 20 rather than 14 sub-divisions within P. trehalosi. The technique demonstrated genetic identity or non-identity among strains of the same or different serotypes from different geographic localities within the UK and was a useful epidemiological tool.

Alleles↗
PubMed1998

Population structure and diversity of avian isolates of Pasteurella multocida from Australia.

A total of 110 isolates of Pasteurella multocida from Australian poultry and reference strains for the 16 somatic serovars plus the three subspecies (gallicida, multocida, septica) were analysed to examine their population structure and diversity. The 81 field isolates examined by multilocus enzyme electrophoresis (MLEE) were diverse, being divided into 56 electrophoretic types (ETs), with the 19 reference strains in another 15 ETs. The population was clonal and somatic serotyping was not particularly useful in establishing relationships between isolates. The 71 ETs formed three distinct subclusters (A, B and C) at a genetic distance of 0.36. Biovars tended to be associated with these subclusters: A with biovars 1, 3, 4, 5 and 8 and B with biovars 2, 6, 7, 9 and 10. Ribotyping, performed on all 110 isolates using Hpall, recognized 21 ribotypes forming nine clusters (R1-R9). The isolates in ribotype cluster R1 were almost identical to those in MLEE cluster B. Using both MLEE and ribotyping, the 19 non-Australian reference strains were found to be distributed over the full diversity of the Australian isolates of P. multocida. This study has shown that a range of P. multocida clones are associated with fowl cholera in Australia and that many of the Australian isolates are similar to non-Australian reference strains. Both the MLEE results and the ribotyping data identified a previously unrecognized subset of P. multocida strains.

Animals↗
PubMed1981

Ovine systemic pasteurellosis caused by Pasteurella haemolytica biotype T.

A detailed study was made of lambs aged 5--7 months naturally infected with Pasteurella haemolytica biotype T. In addition to the well known features of such infections, previously unreported necrotic lesions of the tonsil, oesophagus, pharynx and adjacent areas were consistently seen. Large numbers of P. haemolytica were present in the tonsil, oesophageal lesions, lung, liver and spleen, but few or none in other tissues. The evidence indicated that the disease was not a true septicaemia. It is postulated that P. haemolytica biotype T already present in the tonsils multiplies and invades the adjacent tissues of the upper alimentary tract; groups of organisms from this site enter the blood stream as emboli, most of which lodge in the capillary beds of the lung and liver; rapid multiplication of organisms in these tissues leads to death from the effects of endotoxin.

Animals↗
PubMed1990

Outer membrane proteins of bovine strains of Pasteurella multocida type A and their doubtful role as protective antigens.

Outer membranes were prepared by the Sarkosyl method from 30 strains of Pasteurella multocida and the closely related Taxon 13, which had been isolated from cattle. The patterns of the outer membrane proteins (OMPs) on SDS-PAGE were generally similar to one another, though the four major proteins (a-d) varied somewhat in molecular mass; these patterns allowed the strains to be arranged into 12 groups. Taxon 13 strains and typical P. multocida strains were indistinguishable, both types being found within the same group. Mice were vaccinated with heat-killed bacteria of three strains and challenged with 10 LD50 of homologous and heterologous live bacteria, representing groups based on OMP patterns; the best protection was afforded by strain W674, which protected against nine of the 17 challenge strains; but there was no correlation between protection and PAGE pattern. Pre-vaccination and pre-challenge sera were used in immunoblotting to probe OMPs from protective and non-protective strains. All three vaccines produced antibody to proteins a and d; these proteins appeared to be common to all strains, varying in molecular mass but not in overall antigenic expression. The antibody response to the other two major OMPs appeared to be PAGE-group specific. There was no correlation between protection and the antigen pattern seen by immunoblotting.

Animals↗
PubMed1990

Production of mouse monoclonal antibodies to Pasteurella multocida type A and the immunological properties of a protective anti-lipopolysaccharide antibody.

Eight monoclonal antibodies (MAbs) were produced from mice immunised with whole cells of heat-killed Pasteurella multocida type A which had been cultured under iron-restricted conditions. The MAbs were selected by an enzyme-linked immunosorbent assay (ELISA) in which the antigen consisted of whole bacteria of the immunising strain. Their reactivity was investigated further by immunoblotting, indirect haemagglutination, a complement-mediated bactericidal assay and passive protection of mice. One of the eight MAbs was shown by immunoblotting to react with lipopolysaccharide (LPS), was bactericidal, and completely protected mice against homologous challenge with 10 LD50 of live bacteria. This MAb was selected for further study. Its reaction with LPS of 17 type-A strains and of single strains of types B, D and E was investigated by immunoblotting. Strains that reacted with the anti-LPS MAb in immunoblots were susceptible to its bactericidal activity and gave high ELISA absorbances. Those that did not react were not susceptible to its bactericidal activity and gave low ELISA readings. The relation between bactericidal activity and ELISA absorbance was highly significant (p less than 0.001). Five of the strongly reacting heterologous strains and one non-reacting strain were selected as challenge organisms in a passive protection experiment: only the mice receiving the reacting strains were protected.

Animals↗
PubMed1991

Dermonecrotic toxin production by strains of Pasteurella multocida isolated from man.

Ninety-four clinical isolates of Pasteurella multocida of human origin were tested for dermonecrotic toxin (DNT) production by three methods: dermonecrotic test in guinea-pigs, Vero cell culture cytotoxicity and ELISA. The strains were isolated from patients living in a rural area with widespread intensive pig breeding. Six strains were found to be toxigenic by the three tests. A major protein band of Mr 145 Kda corresponding to DNT on immunoblots was demonstrated in extracts from these strains. All were isolated from respiratory tract (diseases 5, healthy carriage 1). The difference between isolates from the respiratory tract and isolates from wounds inflicted by pets was statistically significant with regard to DNT production (p less than 0.02). A possible role of the toxin in pulmonary diseases caused by P. multocida has yet to be established.

Adult↗
PubMed1998

Demonstration that Australian Pasteurella multocida isolates from sporadic outbreaks of porcine pneumonia are non-toxigenic (toxA-) and display heterogeneous DNA restriction endonuclease profiles compared with toxigenic isolates from herds with progressive atrophic rhinitis.

Capsular types A and D of Pasteurella multocida cause economic losses in swine because of their association with progressive atrophic rhinitis (PAR) and enzootic pneumonia. There have been no studies comparing whole-cell DNA profiles of isolates associated with these two porcine respiratory diseases. Twenty-two isolates of P. multocida from diseased pigs in different geographic localities within Australia were characterised genotypically by restriction endonuclease analysis (REA) with the enzyme CfoI. Seven of 12 P. multocida isolates from nasal swabs from pigs in herds where PAR was either present or suspected displayed a capsular type D phenotype. These were shown to possess the toxA gene by polymerase chain reaction (PCR) and Southern hybridisation, and further substantiated by production of cytotoxin in vitro. The CfoI profile of one of these seven isolates, which was from the initial outbreak of PAR in Australia (in Western Australia, WA), was identical with profiles of all six other toxigenic isolates from sporadic episodes in New South Wales (NSW). The evidence suggests that the strain involved in the initial outbreak was responsible for the spread of PAR to the eastern states of Australia. Another 10 isolates, representing both capsular types A and D, were isolated exclusively from porcine lung lesions after sporadic outbreaks of enzootic pneumonia in NSW and WA. CfoI restriction endonuclease profiles of these isolates revealed considerable genomic heterogeneity. Furthermore, none of these possessed the toxA gene. This suggests that P. multocida strains with the toxA gene do not have a competitive survival advantage in the lower respiratory tract or that toxin production does not play a role in the pathology of pneumonic lesions, or both. REA with polyacrylamide gel electrophoresis and silver staining was found to be a practical and discriminatory tool for epidemiological tracing of P. multocida outbreaks associated with PAR or pneumonia in pigs.

Animals↗
PubMed1999

Genomic DNA restriction site heterogeneity in bovine Pasteurella multocida serogroup A isolates detected with an rRNA probe.

A total of 81 Pasteurella multocida isolates from healthy and diseased dairy and beef cattle originating from various geographical locations was examined by rRNA gene restriction site polymorphism analysis (ribotyping), restriction endonuclease analysis (REA), SDS-PAGE analysis of whole-cell (WCP) and outer-membrane (OMP) proteins, and capsule and somatic serotyping. Bacterial strains were isolated from nose, lung and in one case testicle, of Holstein and cross-bred beef cattle. The isolates represented for the most part serogroup A3 (88%). Ribotyping was performed on DNA digested with HaeII, electrophoresed and then hybridised with 32P-labelled 16S-23S rRNA from Escherichia coli. Six ribotypes (R1-R6) and 10 REA types were found among the 81 isolates with similar discrimination index (DI) of c. 0.60. Protein profiles revealed reproducibility and high levels of polymorphisms among lung isolates. Isolates were compared according to their geographical habitat, their isolation from dairy or from beef cattle and from nasal cavities or lungs. No correlation was apparent between geographical locations and ribotypes. Overall, isolates obtained from dairy cattle were predominantly R1, whereas those obtained from beef cattle were equally distributed between R1 and R2. R1 was more representative of lung isolates. For some strains, particularly the single isolate ribotypes, good correlation was achieved between WCP analysis, REA types and ribotypes. For others, REA to some extent and WCP profiles were able to discriminate among isolates within ribotypes. The data suggest that a combination of ribotyping, REA and WCP analysis is useful for investigating the epidemiology of bovine P. multocida serogroup A.

Animals↗
PubMed2003

Alteration of DNA adenine methylase (Dam) activity in Pasteurella multocida causes increased spontaneous mutation frequency and attenuation in mice.

Pasteurella multocida is one of the primary bacterial pathogens associated with bovine respiratory disease (BRD) complex. Relatively few virulence factors of P. multocida have been characterized, and there is a need for improved vaccines for prevention of BRD. In other Gram-negative species, DNA adenine methylase (Dam) regulates the expression of virulence genes, and appropriate expression of Dam is required for virulence. In this study, the authors cloned and sequenced the P. multocida A1 dam gene and demonstrated that it is able to restore Dam function in an Escherichia coli dam mutant. When P. multocida dam was placed under the control of a constitutively expressed promoter on a plasmid, it caused an increased spontaneous mutation rate in P. multocida. In addition, the plasmid-mediated alteration of Dam production in P. multocida caused it to be highly attenuated in mice. These findings indicate that appropriate expression of Dam is required for virulence of P. multocida, which is believed to be the first report that Dam is required for virulence of a species in the Pasteurellaceae. Therefore, Dam may function as a virulence gene regulator in the Pasteurellaceae, similar to previously reported findings from other Gram-negative species.

Animals↗