Molecular typing of haemorrhagic septicaemia-associated Pasteurella multocida isolates from Pakistan and Thailand using multilocus sequence typing and pulsed-field gel electrophoresis

Moustafa, Ahmed; Bennett, Mark D.; Edwards, John; Azim, Kamran; Mesaik, M. Ahmed; Choudhary, M. Iqbal; Pathanasophon, Pornpen; Worarach, Apasara; Ali, Qurban; Abubakar, Muhammad; Anjum, Rehana

doi:10.1016/j.rvsc.2013.07.003

Cited by 19 publications

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“…multocida strains, isolated from HS cases in buffalo (8 isolates) and cattle (2 isolates) from Pakistan and Thailand, and two Pakistani HS vaccine strains were used for this study ( Table 1 ). Except for the Faisalabad isolate ( Table 1 ), all of the isolates had previously been genotyped using MLST as ST122 [ 14 ]. Therefore, in order to identify if there were differences between the strains, we determined whole genome draft sequences of each strain.…”

Section: Resultsmentioning

confidence: 99%

“…Except for the Faisalabad isolate ( Table 1 ), all of the isolates had previously been identified as P . multocida and typed using MLST (all sequence type 122) [ 14 ]. The Faisalabad isolate was received from the National Veterinary Laboratory, Islamabad, Pakistan, and identified as a HS-associated P .…”

Section: Methodsmentioning

confidence: 99%

“…All isolates were serovar B:2 and indistinguishable by multi locus sequence typing (MLST) with all strains sequence type 122. Furthermore, all isolates from within each country were indistinguishable by pulsed-field gel electrophoresis (PFGE) [ 14 ]. Therefore, to determine whether there was any diversity across these isolates we determined whole draft genome sequences of a selection of 12 of the strains using next generation sequencing (NGS).…”

Section: Introductionmentioning

confidence: 99%

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Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes

et al. 2015

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Pasteurella multocida is the primary causative agent of a range of economically important diseases in animals, including haemorrhagic septicaemia (HS), a rapidly fatal disease of ungulates. There is limited information available on the diversity of P. multocida strains that cause HS. Therefore, we determined draft genome sequences of ten disease-causing isolates and two vaccine strains and compared these genomes using a range of bioinformatic analyses. The draft genomes of the 12 HS strains were between 2,298,035 and 2,410,300 bp in length. Comparison of these genomes with the North American HS strain, M1404, and other available P. multocida genomes (Pm70, 3480, 36950 and HN06) identified a core set of 1,824 genes. A set of 96 genes was present in all HS isolates and vaccine strains examined in this study, but absent from Pm70, 3480, 36950 and HN06. Moreover, 59 genes were shared only by the Asian B:2 strains. In two Pakistani isolates, genes with high similarity to genes in the integrative and conjugative element, ICEPmu1 from strain 36950 were identified along with a range of other antimicrobial resistance genes. Phylogenetic analysis indicated that the HS strains formed clades based on their country of isolation. Future analysis of the 96 genes unique to the HS isolates will aid the identification of HS-specific virulence attributes and facilitate the development of disease-specific diagnostic tests.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes

et al. 2015

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“…Clustering of the 191 isolates analysed, which was based mainly on the profiles of eight genes, showed that the profiles of the Thai bovine isolates of P. multocida were less diverse than those of the porcine isolates. The results were supported by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) of eight field isolates and one vaccine strain of P. multocida associated with HS cases from Thailand [ 43 ]. All isolates shared the same MLST sequence type 122 and the same PFGE pattern, whereas the method used in the current study was able to distinguish additional minor groups of the bovine isolates.…”

Section: Discussionmentioning

confidence: 98%

OmpA protein sequence-based typing and virulence-associated gene profiles of Pasteurella multocida isolates associated with bovine haemorrhagic septicaemia and porcine pneumonic pasteurellosis in Thailand

et al. 2017

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Background Pasteurella multocida is a Gram-negative bacterium that causes economically significant infections of a broad range of animal species. Pneumonic and septicaemic pasteurellosis caused by this bacterium remain important problems in pigs, cattle, and water buffaloes in Thailand. The aim of this study was to characterise the virulence-associated gene profiles and to develop an OmpA molecular typing scheme for classifying 191 bovine and porcine isolates of P. multocida collected between 1989 and 2012 in Thailand using polymerase chain reactions (PCRs), nucleotide sequencing, and sequence and structural bioinformatics analyses.ResultsPCR screening successfully characterised the profiles of 25 virulence-associated genes in all isolates. The gene profiles separated these isolates into bovine and porcine clusters based on eight genes (hgbB, hsf1, tadD, nanH, pfhA, plpE, pmHAS, and tbpA). Phylogenetic analyses of the nucleotide and protein sequences corresponding to the ompA gene, which encodes a major outer membrane surface protein, showed two major bovine and porcine clusters. Structural prediction and analysis of the dN/dS ratio revealed four hypervariable extracellular loops of the OmpA transmembrane domains. These four loops were used to develop an OmpA typing scheme. This scheme classified 186 isolates into five major loop sequence types (LST8, LST12, LST15, LST18, and LST19), consistent with the phylogenetic results. The loop regions of the bovine isolates were predicted to be more antigenic than those of the porcine isolates. Thus, molecular evolution of the OmpA proteins could be used to classify P. multocida isolates into different capsular types, host types, and, possibly, pathogenicity levels.ConclusionsTogether with the virulence-associated gene profiles, the typing reported in this work provides a better understanding of P. multocida virulence. Effective monitoring and potential strain-specific subunit vaccines could be developed based on these loop oligopeptides.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-017-1157-6) contains supplementary material, which is available to authorized users.

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“…ST 122 is predominantly associated with HS cases and has been recovered from Asia and African continent (Hotchkiss et al, 2011;Moustafa et al, 2013). ST 63, ST 162 and ST 147 have been rarely isolated from HS cases, and these STs along with ST 122 have been referred as HS clonal complex (Petersen et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Molecular Epidemiology ofPasteurella multocidaCirculating in India by Multilocus Sequence Typing

Sarangi

Thomas

Gupta

et al. 2014

Transbound Emerg Dis

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Multilocus sequence typing (MLST), a sequence-based typing method for bacterial pathogens, is currently the best method for long-term epidemiological study and to understand the population structure of the bacteria. This investigation was carried out to study the diversity of Pasteurella multocida isolates circulating in India. Ten different sequence types (ST) identified in this study are ST 122 from cattle, goat, mithun and pig; ST 50 from pig; ST 9 from cattle and sheep; ST 229 from cattle and goat; ST 71 and ST 277 from cattle; and ST 129, ST 280, ST 281 and ST 282 from avian species. Of these, ST 277, ST 280, ST 281 and ST 282 were identified for the first time. The analysis of results provides novel epidemiological information on the circulation of multiple STs across India. The majority of STs or their variants identified in this study have already been reported from different parts of the globe. This suggests that probably transboundary spread of strains across countries and continents has occurred across evolutionary time and is still happening. The isolation of ST 122 from small ruminants and pigs suggests that these species may be included in the preventive vaccination policy for effective control of haemorrhagic septicaemia in India.

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Molecular typing of haemorrhagic septicaemia-associated Pasteurella multocida isolates from Pakistan and Thailand using multilocus sequence typing and pulsed-field gel electrophoresis

Cited by 19 publications

References 19 publications

Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes

Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes

OmpA protein sequence-based typing and virulence-associated gene profiles of Pasteurella multocida isolates associated with bovine haemorrhagic septicaemia and porcine pneumonic pasteurellosis in Thailand

Molecular Epidemiology ofPasteurella multocidaCirculating in India by Multilocus Sequence Typing

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