Studies on the presence and persistence of<i>Pasteurella multocida</i>serovars and genotypes in fowl cholera outbreaks

Singh, Reema; Blackall, P. J.; Remington, B. A.

doi:10.1080/03079457.2013.854861

Cited by 27 publications

(32 citation statements)

References 24 publications

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“…As an example, LPS PCR result L3 should be either serovar 3 or serovar 4, yet the Heddleston serotyping of these isolates gave results such as NT, 14/15 cross‐reacting, 13/15/16 cross‐reacting, 4/14 cross‐reacting and 1/3 cross‐reacting. This lack of correlation was also observed when the LPS PCR was developed and in previous application of the PCR . In the original development of the PCR, the gold standard method of a full chemical and structural analysis showed that LPS PCR was consistently correct, while the traditional serotyping methodology was in error, when these two tests were in disagreement.…”

Section: Discussionmentioning

confidence: 99%

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Genotypic diversity of Pasteurella multocida isolates from pigs and poultry in Australia

2018

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Objective To investigate the genotype and diversity of Pasteurella multocida present in pig herds and to determine the extent of overlap with isolates from poultry flocks in Australia. Methods A total of 43 isolates from pigs from different farms and regions of Australia were used in this study. A diverse collection of 41 poultry isolates, with 31 being previously characterised, was also used. The pig isolates and 10 poultry isolates were identified by species‐specific PCR assay, serotyped by the Heddleston scheme and genotyped by a multiplex PCR based on the lipopolysaccharide (LPS) outer core biosynthesis locus, repetitive element PCR fingerprinting (rep‐PCR) and multilocus sequence typing (MLST), with the latter being used on a subset of the isolates based on the rep‐PCR results. Results Only 4 out of 8 recognised LPS genotypes were found in the pig isolates, with each isolate assigned to an LPS genotype. In contrast, 77% of the isolates were non‐typable or cross‐reacting in the Heddleston serotyping scheme. The rep‐PCR analysis recognised 20 patterns, yet only 16 sequence types (STs) were found and 4 were new STs. There were 5 STs (STs 7, 11, 20, 24 and 58) shared among the pig and poultry isolates. Conclusions Although only limited numbers of isolates have been examined, there is evidence of a sharing of genotypes between Australian pigs and chickens. These findings have major implication for biosecurity measures with regard to minimising both direct (e.g. animal to animal) and in‐direct (e.g. shared staff or cross‐visitors) contact between poultry and pigs.

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Section: Discussionmentioning

confidence: 99%

“…Similarly, the diversity of Australian pig isolates has been examined in terms of phenotypic properties, as well as by genotypic methods such as restriction endonuclease analysis, ribotyping and plasmid analysis . Multilocus sequence typing (MLST) has proven useful in two Australian‐based studies looking at fowl cholera outbreaks …”

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Genotypic diversity of Pasteurella multocida isolates from pigs and poultry in Australia

2018

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“…In this case, an autogenous vaccine should include all relevant genotypes or serovars in the outbreak (SINGH et al, 2013).…”

Section: Resultsmentioning

confidence: 99%

Molecular characterization of Pasteurella multocida isolates from swine lungs by Randomly Amplified Polymorphic DNA

et al. 2015

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“…However, the accuracy of Heddleston serotyping has never been objectively tested, as the precise LPS structures produced by different strains have not been known. Indeed, there have been many informal as well as formal reports that the Heddleston system fails to type many isolates and lacks accuracy and reproducibility (7,8). Furthermore, Heddleston serotyping is time-consuming and requires access to good-quality, serovar-specific antisera.…”

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Development of a Rapid Multiplex PCR Assay To Genotype Pasteurella multocida Strains by Use of the Lipopolysaccharide Outer Core Biosynthesis Locus

et al. 2015

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Pasteurella multocida is a Gram-negative bacterial pathogen that is the causative agent of a wide range of diseases in many animal species, including humans. A widely used method for differentiation of P. multocida strains involves the Heddleston serotyping scheme. This scheme was developed in the early 1970s and classifies P. multocida strains into 16 somatic or lipopolysaccharide (LPS) serovars using an agar gel diffusion precipitin test. However, this gel diffusion assay is problematic, with difficulties reported in accuracy, reproducibility, and the sourcing of quality serovar-specific antisera. Using our knowledge of the genetics of LPS biosynthesis in P. multocida, we have developed a multiplex PCR (mPCR) that is able to differentiate strains based on the genetic organization of the LPS outer core biosynthesis loci. The accuracy of the LPS-mPCR was compared with classical Heddleston serotyping using LPS compositional data as the "gold standard." The LPS-mPCR correctly typed 57 of 58 isolates; Heddleston serotyping was able to correctly and unambiguously type only 20 of the 58 isolates. We conclude that our LPS-mPCR is a highly accurate LPS genotyping method that should replace the Heddleston serotyping scheme for the classification of P. multocida strains. P asteurella multocida is the primary causative agent of a wide range of economically important diseases, including hemorrhagic septicemia in ungulates, atrophic rhinitis in pigs, fowl cholera in birds, snuffles in rabbits, and enzootic pneumonia and shipping fever in cattle, sheep, and pigs (1). P. multocida also causes opportunistic infections in humans, often following cat or dog bites, and plays a contributory role, together with other pathogens, in a range of lower respiratory tract infections and sporadic septicemias in ungulates (1).P. multocida strains have classically been differentiated using serological techniques. Strains can be classified into five capsular serogroups (A, B, D, E, and F) using an indirect hemagglutination test (2) and into 16 somatic or lipopolysaccharide (LPS) serovars (serotypes) using the Heddleston gel diffusion precipitin test (3). Both of these schemes have been widely used. Isolates are commonly assigned a combined designation, such as A:1 (capsular serogroup A and LPS serovar 1) or B:2 (capsular serogroup B and LPS serovar 2).P. multocida LPS is an immunodominant antigen critical for homologous protection stimulated by bacterin (killed-cell) vaccines (4). Furthermore, in the P. multocida strain VP161, a fulllength LPS molecule is essential for the ability to cause acute disease (5, 6). Heddleston serotyping is currently the only method used to differentiate P. multocida strains on the basis of LPS type. However, the accuracy of Heddleston serotyping has never been objectively tested, as the precise LPS structures produced by different strains have not been known. Indeed, there have been many informal as well as formal reports that the Heddleston system fails to type many isolates and lacks accuracy and reproducibilit...

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Studies on the presence and persistence ofPasteurella multocidaserovars and genotypes in fowl cholera outbreaks

Cited by 27 publications

References 24 publications

Genotypic diversity of Pasteurella multocida isolates from pigs and poultry in Australia

Genotypic diversity of Pasteurella multocida isolates from pigs and poultry in Australia

Molecular characterization of Pasteurella multocida isolates from swine lungs by Randomly Amplified Polymorphic DNA

Development of a Rapid Multiplex PCR Assay To Genotype Pasteurella multocida Strains by Use of the Lipopolysaccharide Outer Core Biosynthesis Locus

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