A phenotypic characterisation of 150 isolates of bacteria previously identified as Pasteurella multocida was performed. All the isolates had been obtained from Australian pigs in the three eastern States of Queensland (110 isolates), New South Wales (21 isolates) and Victoria (19 isolates). Seven different biochemical biovars were recognised amongst the isolates. A total of 100 isolates (67%) were assigned to biovar 3, previously shown to be the most common biovar in isolates of P. multocida from Australian poultry [Fegan, N., Blackall, P.J., Pahoff, J.L., 1995. Phenotypic characterisation of Pasteurella multocida isolates from Australian poultry. Vet. Microbiol., 47, 281-286.]. Six of the seven biovars, including biovar 3, were identified as P. multocida subsp. multocida, 124 isolates in total. One other biovar, consisting of thirteen isolates, was identified as P. multocida subsp. gallicida. Within the six biovars that were identified as P. multocida subsp. multocida, biovars 12, 13 and 14 represented unusual biochemical variants. The nine isolates assigned to biovar 12 appeared to be lactose positive variants of P. multocida subsp. multocida. The three isolates in biovar 13 appeared to be ornithine decarboxylase (ODC) negative variants of P. multocida subsp. multocida. The single isolate in biovar 14 appeared to be an ODC negative, lactose positive variant of P. multocida subsp. multocida.

Both of the vaccines provided significant protection against a severe challenge with serovar 1 A pleuropneumoniae. Neither vaccine was effective against a serovar 15 A pleuropneumoniae challenge. There was evidence that the Porcilis APP vaccine did provide some protection against the serovar 15 challenge because the ADG, after challenge of pigs given this vaccine, was greater than the control pigs.

Objective To use the technique of ribotyping to investigate the genetic diversity of Australian isolates of Pasteurella multocida associated with outbreaks of clinical disease in Australian pigs. Design One hundred and seven porcine P multocida isolates were analysed by ribotyping using the restriction enzymes HpaII and HindIII. The genetic population structure of the Australian porcine P multocida isolates was determined through statistical analysis of the joint ribotype patterns, and this was then compared with biochemical and epidemiological data available for the population. Results A total of 25 combined ribotypes were recognised, which were grouped into five ribotype clusters. Despite the deliberate selection of diverse isolates, the study revealed only a limited degree of genetic diversity. Fourteen of the ribotypes contained multiple isolates, and 12 of these ribotypes were present on more than one farm. Three of the seven biovars analysed in the study showed very limited diversity. All fifteen biovar 2 isolates (subsp multocida) were found in a single cluster (III), while all four biovar 8 isolates, which correspond to P multocida subsp gallicida, were allocated by themselves to a single cluster (IV). All nine of the biovar 12 isolates (lactose‐positive subsp multocida) were assigned to a single cluster (I), together with the single biovar 14 isolate, which was the only other lactose‐positive isolate in the population (ODC‐negative). Conclusion A limited number of ribotypes of P multocida are associated with Australian pigs. The majority of these ribotypes are widely distributed across multiple farms, and across multiple states. Individual farms can possess multiple ribotypes of P multocida. Some of the unusual biochemical variants of P multocida present in Australian pigs have a very limited genetic diversity. The nature of pig production in Australia, primarily involving continuous flow systems with few closed herds, has possibly contributed to the widespread distribution of a limited number ribotypes.

Two outbreaks of fowl cholera on a multiage free-range egg farm were investigated. The outbreaks occurred in 1994 and 2002. A total of 22 strains of Pasteurella multocida were available for study, 11 from the 1994 outbreak and 11 from the 2002 outbreak. Lesions typical of acute fowl cholera were seen in the 1994 outbreak, whereas both acute and chronic fowl cholera occurred in the 2002 outbreak. The isolates were examined in an extended phenotypic typing methodology, by a P. multocida–specific polymerase chain reaction (PCR), by the Heddleston somatic serotyping scheme, and by restriction endonuclease analysis (REA) typing using the enzyme HpaII. All 22 strains had the same phenotypic properties, all were confirmed as P. multocida by PCR, all were Heddleston serovar 4, and all had the same REA pattern. The results indicate that these 2 outbreaks were caused by the same clone of P. multocida–-despite the 8-year time period between the outbreaks.

Objectives To clarify the serological identity of the proto type strain of a group of Actinobacillus pleuropneumoniae isolates that could not be serotyped in previous studies and to establish the serovar of 378 isolates of A pleuropneumoniae obtained from pigs in Australia over the period 1993 to 1996.Design After initial validation, QGD and IHA tests were used to characterise the prototype isolate (HS143) selected to represent the cross‐reacting isolates that were found in a previous study. Next, 378 recent field isolates of A pleurop‐neumoniae were characterised using the existing gel diffusion serotyping technique and/or the IHA or QGD tests.Results The indirect haemagglutination test was shown to be capable of correctly recognising the reference strain for all serovars except serovar 11. While the quantitative gel diffu sion test was not as effective as indirect haemagglutination, it could recognise serovar 11. When the two tests were used to examine the prototype strain (HS143) of the cross‐reactive isolates, the results indicated that HS143 is serologically distinct from all 12 of the recognised serovars of A pleurop‐neumoniae. However, as HS143 was subsequently identified as serovar 12 by one of the leading international reference laboratories, the antiserum to isolate HS143 was used as the serovar 12 antiserum. A total of 346 of the 378 A pleuropneumoniae field isolates examined could be confidently serotyped with almost 90% of the isolates being either serovar 1 (104 isolates); serovar 7 (83 isolates) or serovar 12 (142 isolates). A range of other serovars and some cross‐reactive isolates made up the remainder of the isolates.Conclusion The serovar 12 antiserum produced against the international reference strain (1096) does not recognise Australian serovar 12 isolates. The antiserum raised against isolate HS143 does recognise the Australian serovar 12 isolates. The dominant serovars of A pleuropneumoniae infecting Australian pigs are (in decreasing order) serovars 12, 1 and 7.

Objective To evaluate the serological response of pigs receiving either the Porcilis APP vaccine or a modified live vaccine based on a streptomycin‐dependent (SD) strain of Actinobacillus pleuropneumoniae, and then challenged with an Australian isolate of A pleuropneumoniae of either serovar 1 or 15 as a means of understanding the protection provided by both vaccines against serovar 1 but not against serovar 15. Design The serological tests evaluated were serovarspecific polysaccharide ELISA tests (for serovar 1 and 15), ELISA tests for antibodies to three A pleuropneumoniae toxins (ApxI, ApxII and ApxIII) as well as to a 42 kDa outer membrane protein (OMP), a haemolysin neutralisation (HN) assay and immunoblotting. The tests were used to detect antibodies in vaccinated pigs that had been shown to be protected against serovar 1 but not serovar 15. Results In the polysaccharide antigen ELISA assays, both vaccines resulted in a significant rise in the titre in the serovar 1 ELISA but not the serovar 15 ELISA. The Porcilis APP vaccinated pigs showed a significant response in the ApxI, ApxIII and 42 kDa OMP ELISA. In the ApxII ELISA, all pigs tested (the Porcilis APP vaccinates and the controls) were positive on entry to the trial. In the HN assay, the Porcilis APP vaccinated pigs showed a significant response after one dose while the SD vaccinated pigs required two doses of vaccine before a marked rise in titre was induced. Immunoblotting revealed that neither vaccine generated antibodies that recognised the ApxIII produced by serovar 15. Conclusions The failure of these vaccines to provide protection against serovar 15 may be due to novel virulence factors possessed by serovar 15, significant differences between the ApxIII toxin of serovar 15 and those present in the Porcilis APP vaccine or failure by both vaccines to induce antibodies to the serovar 15 specific polysaccharide.

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