research article virulence genotyping of pasteurella multocida...

Research ArticleVirulence Genotyping of Pasteurella multocidaIsolated from Multiple Hosts from India

Laxmi Narayan Sarangi12 Adyasha Priyadarshini1 Santosh Kumar1 Prasad Thomas13

Santosh Kumar Gupta1 Viswas Konasagara Nagaleekar1 and Vijendra Pal Singh1

1 Division of Veterinary Bacteriology and Mycology Indian Veterinary Research Institute Izatnagar BareillyUttar Pradesh 243122 India

2 Regional Medical Research Centre Bhubaneswar Odisha 751023 India3 Institute of Bacterial Infections and Zoonoses Friedrich Loeffler Institut Namburger Straszlige 96 a 07743 Jena Germany

Correspondence should be addressed to Santosh Kumar Gupta vetsantoshgmailcom

Received 30 July 2014 Revised 25 September 2014 Accepted 17 October 2014 Published 18 November 2014

Academic Editor Hela Kallel

Copyright copy 2014 Laxmi Narayan Sarangi et alThis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

In this study 108 P multocida isolates recovered from various host animals such as cattle buffalo swine poultry (chicken duckand emu) and rabbits were screened for carriage of 8 virulence associated genes The results revealed some unique information onthe prevalence of virulence associated genes among Indian isolates With the exception of toxA gene all other virulence associatedgenes were found to be regularly distributed among host species Association study between capsule type and virulence genessuggested that pfhA nanB and nanH genes were regularly distributed among all serotypes with the exception of CapD whereastoxA gene was found to be positively associated with CapD and CapAThe frequency of hgbA and nanH genes among swine isolatesof Indian origin was found to be less in comparison to its equivalents around the globe Interestingly very high prevalence of tbpAgene was observed among poultry swine and rabbit isolates Likewise very high prevalence of pfhA gene (953) was observedamong Indian isolates irrespective of host species origin

1 Introduction

Pasteurella multocida belonging to family Pasteurellaceae is aubiquitous organism affectingmultihost species thus causingseveral diseases like haemorrhagic septicaemia in cattle andbuffalo enzootic bronchopneumonia in cattle sheep andgoats atrophic rhinitis in swine fowl cholera in poultryand snuffles in rabbits [1 2] These diseases are known tocause severe financial loss to livestock industry especially intropical countries Conventional vaccines have been used forseveral decades as a control strategy but major limitation ofthese vaccines is their ineffectiveness in inducing long actingcross protective immunity [3 4] Therefore several outermembrane proteins (OMPs) have been proposed as candidateantigen for subunit vaccine [3 5]

The OMPs of Gram negative bacteria play an essentialrole in the disease process They are involved in the process

of nutrient uptake transport of molecules in and out ofthe cell colonization and invasion of the host evasion ofhost immune response injury to host tissue and so forthrequired for productive infection [6] These proteins aresubjected to different selection pressures thereby exhibitingvarying degree of interstrain heterogeneity Therefore thesevirulence associated genes can be used to assess intraspeciesdiversity and also to obtain epidemiological relationships [7]In addition these OMPs are good immunogens and can beused as vaccine components to provide protection [8ndash10]Hence virulence profiling can be used as a typing methodfor characterization of bacterial pathogens [11] and also fordevelopment of subunit vaccine in vaccine strain selectionFor the first time virulence profiling of P multocida isolateswas carried out by Ewers et al [12] and subsequently it hasbeen used bymany workers to understand the diversity of thepathogen recovered from different host origin [13ndash19]

Hindawi Publishing Corporatione Scientific World JournalVolume 2014 Article ID 814109 10 pageshttpdxdoiorg1011552014814109

2 The Scientific World Journal

The previous study carried out in our laboratory on car-riage of 19 virulence genes among Pmultocida isolates recov-ered from small ruminants revealed some novel informationon the frequency of virulence genes like very high prevalenceof pfhA gene 489 prevalence of toxA gene with the highestprevalence among serotype A followed by serotype D andin one isolate each of capsular types B and F (Sarangi etal submitted for publication) These findings from smallruminant isolates encouraged sampling of more isolates ofIndian origin from various hosts to have a clear under-standing on the heterogeneity of the bacteria Therefore thisstudy was extended to P multocida isolates recovered frommultiple host species for 8 important virulence associatedgenes encoding proteins involved in bacterial survival andpathogenesis It included genes encoding transferrin bindingprotein (TbpA) and haemoglobin binding protein (HgbAHgbB) associated with iron acquisition filamentous haemag-glutinin (PfhA) subunit of type IV fimbriae (PtfA) sialidases(NanB NanH) involved in initial colonization and adhesionand dermonecrotoxin (ToxA)

2 Materials and Methods

21 Bacterial Strains In the present study 108 P multocidaisolates recovered from large ruminants (buffalo 119899 = 23cattle 119899 = 18) avians (chicken 119899 = 18 duck 119899 = 8 andemu 119899 = 4) swine (119899 = 34) and rabbit (119899 = 3) maintainedat Division of Bacteriology amp Mycology Indian VeterinaryResearch Institute Izatnagar were used Selections of isolateswere carried out on the basis of host origin year andplace of isolation in order to incorporate isolates from allover India The details of the isolates (isolate number hostorigin capsular type year and place of isolation and diseasesymptom (if available)) are given in Table 1

22 Confirmation of P multocida Isolates The isolates wererevived in brain heart infusion broth by 18ndash24 h incubationat 37∘C and plated subsequently onto blood agar to study cul-tural characteristics The cultures were then tested for purityby biochemical tests as per standard techniques [20] Thegenomic DNA of the isolates was extracted by CTABmethod[21] and the isolates were reconfirmed as Pmultocida by PM-PCR followed by determination of capsular type bymultiplexPCR [22 23]

23 Detection of Virulence Associated Genes by PCR Theisolates were then subjected to screening of 8 virulence genesencoding iron binding proteins (TbpA HgbA HgbB) colo-nization and adhesion related protein (PfhA PtfA) sialidases(NanB NanH) and dermonecrotoxin (ToxA) by individualPCR reactions utilizing oligonucleotide primers describedpreviously The details of the virulence genes sequences ofthe oligonucleotide primers and citations used are listed inTable 2

24 Statistical Analysis Statistical analysis of the data gener-ated from the study was performed with SPSS 160 (SPSS IncChicago) 119875 values of lt005 were considered as statisticallysignificant

3 Results and Discussion

P multocida is an economically important veterinarypathogen causing wide range of diseases in livestock andpoultry The bacteria have been classified into five capsulartypes (A B D E and F) based on capsular typing witheach capsule type being predominantly associated with aparticular disease in a host species But isolation of othercapsular types from such hosts by cross species infection isnot uncommon [2 7] Ability of the bacteria to infect andsurvive in several hosts as commensal exposes it to variousselection pressures resulting in emergence of divergentstrains in field scenario Molecular epidemiological studyby employing REP-PCR ERIC-PCR MLST analysis and soforth has confirmed the diversity of P multocida circulatingin India and also the possibility of transboundary spreadof strains across evolutionary time [24 25] Therefore adetailed study on the presence of virulence associated genesrecovered from different host species in Indian subcontinentwill be helpful to understand the disease process and todevelop disease control measures in future

In this study P multocida isolates recovered from varioushost species were screened for presence of 8 importantvirulence associated genes (tbpA hgbA hgbB pfhA ptfAnanB nanH and toxA) involved in bacterial pathogenesisThe results confirmed that with the exception of toxA geneall other virulence associated genes are regularly distributedamong the isolates of different host origin The result ofindividual PCR reaction for each isolate is presented inTable 1 Among the genes encoding iron binding proteinstbpA gene was present in 824 of isolates which range from696 in buffalo to 100 in cattle emu and rabbits (Table 3)Similarly hgbA gene was found to be regularly distributedamong all isolates affecting different hosts with the lowestprevalence among swine isolates (735) Gene hgbB hasthe lowest prevalence among the three iron binding proteinsscreened in this study and was found in 722 of the isolatesThe percentage prevalence of this gene was found to be moreamong avian isolates (90) in comparison to large ruminants(659) and swine (676) isolates Of the two sialidasespresent in P multocida isolates the percentage prevalence ofnanB gene was found to be more than nanH gene Overallvery high prevalence of pfhA gene (935) was observedin this study with 100 prevalence among avian isolatesDermonecrotoxin gene (toxA) was found only in 176 ofstrains with majority of the isolates belonging to porcineorigin One buffalo and 3 duck isolates were also found tocarry toxA gene (Table 3)

Iron acquisition and uptake are essential for bacterialsurvival and many bacteria have developed different ironsequestering system for uptake of ironThe expression of ironacquisition proteins increases under iron limiting conditionas well as in vivo condition (reviewed in [5]) P multocidautilizes various receptors for adapting to variations in supplyof different haem iron sources [5] Among these TbpAprotein is necessary for extraction of iron from transferrinand has been reported to be an important virulence factorand epidemiological marker in cattle [12 19 26] Previousstudies reported that tbpA gene is either absent or rarely

The Scientific World Journal 3

Table 1 Details of the isolates (host origin serotype detected place of isolation year of isolation symptom presenceabsence of individualvirulence genes)

Sample id Species Serotype Place Year Diseasesymptom tbpA hgbA hgbB pfhA ptfA toxA nanB nanH10 Cattle F Pune 1992 NA P P P P P A P P11 Cattle F Pune 1992 NA P P P P P A P P51 Pig A UP 1995 NA P P P P P P P A53 Buffalo B Pune 1996 NA A P P P P A P A98 Duck A Tripura 2001 NA P P P P P A P P117 Cattle B Bhubaneswar 2001 NA P P P P P A P P118 Cattle B Bhubaneswar 2001 NA P P P P P A P P120 Cattle B Bhubaneswar 2001 NA P P P P P A P P128 Cattle B Bangalore 2001 NA P P P P P A P P132 Buffalo A Palampur 2001 NA P P P P P A P P133 Buffalo A Palampur 2001 NA A A P P P A P A134 Buffalo A Palampur 2001 NA P P P P P A P P141 Chicken A Chennai 2001 NA P P P P P A P P202 Chicken A Chennai 2002 NA A P P P P A P A206 Chicken B Chennai 2002 NA P P P P P A P A222 Buffalo A Mathura 2002 NA P P A P P A P P258 Chicken A Nasik 2002 NA P P P P P A P P330 Chicken B Anand 2002 NA P P P P P A P P288 Buffalo B Bhubaneswar 2003 NA P P P P P A P P291 Pig B Guwahati 2003 NA P P A P P A P P292 Pig B Guwahati 2003 NA P P P P P A P A366 Cattle B Palampur 2004 NA P A A P P A P P390 Buffalo B Palampur 2005 NA P P P P P A P P400 Buffalo B Ludhiana 2005 NA P P P P P A P P407 Chicken B Ludhiana 2005 NA P P P P P A P P409 Buffalo B Jammu 2005 NA P P P P P A P P410 Buffalo B Jammu 2005 NA P P P P P A P P425 Duck B Chennai 2005 NA P P P P P P P P448 Rabbit B Palampur 2006 NA P P A P P A P P456 Chicken A Chennai 2006 NA P P P P P A P P460 Chicken A Chennai 2006 NA A A A P P A P A464 Chicken A Chennai 2006 NA A P P P P A P P569 Chicken A Chennai 2007 NA P A A P P A P A537 Pig A Guwahati 2007 NA A A A P P A A P540 Pig A Guwahati 2007 NA P P P P P P P P543 Pig D Guwahati 2007 NA P P P P P P P A550 Duck A Guwahati 2007 NA P A P P P P P P555 Buffalo B Anand 2007 NA P P A P P A P A559 Rabbit B Palampur 2007 Nasal discharge P P A P P A P P563 Cattle B Anand 2007 NA P P A P P A P P585 Pig A Guwahati 2008 NA P P A P P P P P587 Pig A Guwahati 2008 NA P P P P P P P P602 Buffalo B Palampur 2008 NA P P P P P A P P608 Rabbit A Palampur 2008 Nasal discharge P P P P P A P P610 Buffalo B Ludhiana 2008 NA A A A P P A P A618 Chicken B Palampur 2008 NA P P P P P A P P


Table 1 Continued

Sample id Species Serotype Place Year Diseasesymptom tbpA hgbA hgbB pfhA ptfA toxA nanB nanH632 Buffalo B Anand 2008 NA A A A P P A P P633 Chicken B Bangalore 2008 NA P A P P P A P P655 Buffalo D Guwahati 2008 NA P P P P P A P P701 Pig A Guwahati 2009 NA P P A P P P P P702 Pig A Guwahati 2009 NA P P P P P A P P703 Pig D Guwahati 2009 NA P P P P P P P A704 Cattle B Guwahati 2009 NA P P A A P A P P720 Pig B UP 2009 NA P P P P P A P P721 Pig B UP 2009 NA P P P P P A P P722 Pig B UP 2009 NA P P A P P P P P725 Buffalo A Ludhiana 2009 NA P P P P P P P A733 Pig D Guwahati 2009 NA P A P P P P P A736 Pig D Guwahati 2009 NA P P P A P A P A737 Pig D Guwahati 2009 NA P P P A P A P A749 Cattle A Palampur 2009 NA P P P P P A P P746 Cattle A Palampur 2009 NA P P P P P A P P747 Cattle A Palampur 2009 NA P P P P A A P P754 Cattle A Palampur 2009 NA P P P P P A P P782 Chicken A Anand 2009 NA A P A P P A P A

794 Chicken A Thrissur 2009 Necrotic foci in liver andhaemorrhage in heart

P P P P P A P A

652 Buffalo B Guwahati 2010 NA P P P P P A P P653 Buffalo B Guwahati 2010 NA P P P P P A P P784 Chicken A Anand 2010 NA P P P P P A P P803 Chicken A Chennai 2010 NA P P P P P A P P804 Chicken A Anand 2010 NA P P P P P A P P811 Cattle A Palampur 2010 NA P P P P P A P P852 Pig A Guwahati 2011 Diseased P P P P P P P P860 Pig B Guwahati 2011 Diseased P P P P P A P P876 Pig A Thrissur 2011 Fever A A A P P A A P877 Pig A Thrissur 2011 Fever A A P P P A P P879 Pig A Thrissur 2011 Fever A P A P P A P A890 Emu A Chennai 2011 NA P P P P P A P P2751 Cattle B Palampur 2011 Nasal discharge P P A P P A P P2766 Cattle B Palampur 2011 Nasal discharge P P P P P A P P3324 Cattle B Palampur 2011 Nasal discharge P P A P P A P P4312 Cattle B Palampur 2011 Nasal discharge P P A P P A P PBP23 Pig B Guwahati 2011 NA P A A P P A P PBP28 Pig A Guwahati 2011 NA P A P P P P P PBP37 Pig A Guwahati 2011 NA P P A P P P P PEMU 2 Emu A Chennai 2011 NA P P P P P A P PJP18 Pig A Guwahati 2011 NA P P P P P P P PNP23 Pig B Guwahati 2011 NA P P P P P A P PNP37 Pig B Guwahati 2011 NA P P P P P A P PPP1A Pig A Thrissur 2011 NA P P P P P A P PPP2A Pig A Thrissur 2011 NA P P P P P P P PPP4A Pig A Thrissur 2011 NA P P P P P A P P


Table 1 Continued

Sample id Species Serotype Place Year Diseasesymptom tbpA hgbA hgbB pfhA ptfA toxA nanB nanH914 Duck A Thrissur 2012 NA P P P P P A P P920 Emu A Anand 2012 NA P P P P P A P P922 Emu A Anand 2012 NA P P P P P A P PDP53 Duck A Thrissur 2013 NA P P P P P A P ADP54 Duck A Thrissur 2013 NA P P P P P P P ADP55 Duck A Thrissur 2013 NA A P P P P A P ADP56 Duck A Thrissur 2013 NA P P P P P A P AP14 Pig D Guwahati 2013 NA A A A A P P A AP15 Pig A Guwahati 2013 NA A A A A P A P AP16 Pig D Guwahati 2013 NA A A P A P A A APAB 78 Buffalo B Anand 2013 NA P P A P P A P PPAB 80 Buffalo B Anand 2013 NA P P A P P A P PPAB 86 Buffalo B Anand 2013 NA A A A A A A P PPAP 88 Chicken A Anand 2013 NA P P P P P A P ALDHB 106 Buffalo B Ludhiana 2014 NA A P A P P A P PMSRB 108 Buffalo B Ludhiana 2014 NA A P P P P A P P(NA = not available A = absence of virulence gene P = presence of virulence gene as detected in PCR reaction)

Table 2 Details of primers and citations used for the detection of capsular type and virulence associated genes in strains of Pasteurellamultocida

Gene Primer Primer sequence (51015840-31015840) ReferencePM-PCR and Capsular serotypes

KMT1 PMPCR-FPMPCR-R

ATCCGCTATTTACCCAGTGGGCTGTAAACGAACTCGCCAC [22]

hyaD-hyaC capA FcapA R

GATGCCAAAATCGCAGTCAGTGTTGCCATCATTGTCAGTG [23]

bcbD capB FcapB R

CATTTATCCAAGCTCCACCGCCCGAGAGTTTCAATCC [23]

dcbF capD FcapD R

TTACAAAAGAAAGACTAGGAGCCCCATCTACCCACTCAACCATATCAG [23]

ecbJ capE FcapE R

TCCGCAGAAAATTATTGACTCGCTTGCTGCTTGATTTTGTC [23]

fcbD capF FcapF R

AATCGGAGAACGCAGAAATCAGTTCCGCCGTCAATTACTCTG [23]

Iron acquisition genes

tbpA tbpA FtbpA R

GGACAGTGCATATAACTTGTTGGACAGTGCATATAACTTGTTTACTA [32]

hgbA hgbA FhgbA R

CATATCGGATCCTTGAAACCAGAGGAAGCAAAAAGAATCGGAGCTCACGACCTGGTGAGTAAAAACGAT In-house [33]

hgbB HgbB FHgbB R

ACCGCGTTGGAATTATGATTGCATTGAGTACGGCTTGACAT [12]

Adhesins

ptfA ptfA FptfA R

AGGATCCATGAAAAAAGCCATTTGGAGCTCTTATGCGCAAAATCCTG In-house

pfhA pfhA FpfhA R

TAAGCCTATCGGTTCAAGTCGGATAAATCTACCCCGTCCTCT In-house

Sialidases

NanB NanB FNanB R

GTCCTATAAAGTGACGCCGAACAGCAAAGGAAGACTGTCC [12]

nanH nanH FnanH R

CACTGCCTTATAGCCGTATTCAGCACTGTTACCCGAACCC [12]

Dermonecrotoxin

ToxA ToxA FToxA R

TCTTAGATGAGCGACAAGGGAATGCCACACCTCTATAG [34]


Table 3 Prevalence of virulence associated genes among Pasteurella multocida isolates recovered from various host species of India

Host origincapsular type No of strains tbpA()

hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()

Buffalo 23 696 826 652 957 957 100 783 43Cap type A 5 800 800 800 100 100 100 600 200Cap type B 17 647 823 588 941 941 100 823 00Cap type D 1 100 100 100 100 100 100 100 00Cap type F 0 NA NA NA NA NA NA NA NACattle 18 100 944 667 944 944 100 100 00Cap type A 5 100 100 100 100 800 100 100 00Cap type B 11 100 909 454 909 100 100 100 00Cap type D 0 NA NA NA NA NA NA NA NACap type F 2 100 100 100 100 100 100 100 00Large ruminants (cattle + buffalo) 41 829 878 659 951 951 100 878 24Cap type A 10 900 900 900 100 900 100 800 100Cap type B 28 785 857 535 928 964 100 892 00Cap type D 1 100 100 100 100 100 100 100 00Cap type F 2 100 100 100 100 100 100 100 00Chicken 18 778 833 833 100 100 100 611 00Cap type A 13 692 846 769 100 100 100 538 00Cap type B 5 100 800 100 100 100 100 800 00Cap type D 0 NA NA NA NA NA NA NA NACap type F 0 NA NA NA NA NA NA NA NADuck 8 875 878 100 100 100 100 500 375Cap type A 7 857 857 100 100 100 100 428 285Cap type B 1 100 100 100 100 100 100 100 100Cap type D 0 NA NA NA NA NA NA NA NACap type F 0 NA NA NA NA NA NA NA NAEmu 4 100 100 100 100 100 100 100 00Cap type A 4 100 100 100 100 100 100 100 00Cap type B 0 NA NA NA NA NA NA NA NACap type D 0 NA NA NA NA NA NA NA NACap type F 0 NA NA NA NA NA NA NA NAAvian (chicken + duck + emu) 30 833 867 900 100 100 100 633 100Cap type A 24 791 875 875 100 100 100 583 83Cap type B 6 100 833 100 100 100 100 833 166Cap type D 0 NA NA NA NA NA NA NA NACap type F 0 NA NA NA NA NA NA NA NARabbit 3 100 100 333 100 100 100 100 00Cap type A 1 100 100 100 100 100 100 100 00Cap type B 2 100 100 00 100 100 100 100 00Cap type D 0 NA NA NA NA NA NA NA NACap type F 0 NA NA NA NA NA NA NA NAPig 34 794 735 676 853 100 882 676 441Cap type A 18 722 722 611 944 100 888 833 442Cap type B 9 100 888 666 100 100 100 888 111Cap type D 7 714 571 857 428 100 714 00 571Cap type F 0 NA NA NA NA NA NA NA NA


Table 3 Continued


hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()

Total (all isolates) 108 824 833 722 935 981 963 750 176Cap type A 53 792 830 792 980 980 962 717 245Cap type B 45 867 867 600 956 978 100 889 44Cap type D 8 750 625 875 500 100 750 125 500Cap type F 2 100 100 100 100 100 100 100 00

present in poultry swine and rabbit isolates (Table 4) [12ndash16] In contrast to these findings we observed a very highoccurrence of tbpA gene among poultry (833) swine(794) and rabbit (100) isolates (Tables 3 and 4) Amongthe host species the prevalence of this gene was found tobe lowest (696) in buffalo (Table 3) The difference inprevalence of this gene among isolates of cattle and buffaloorigin was found to be statistically significant (119875 lt 005)which is quite unexpectedThereforemore number of isolatesfrom both host origins should be carried out before reachingany conclusion In this study tbpA gene was found to befrequently distributed among four capsule types includingCapF (Table 3) This is in contrast to Ewers et al [12] whoobserved tbpA gene in 70 of CapB strains followed by37 of CapA 95 of CapD strains and nil in CapF strainsSimilarly Katsuda et al [18] reported positive association ofCapA strain with tbpA gene

P multocida utilizes two proteins (HgbA and HgbB) foracquiring iron directly from haem component Morton etal [27] reported that the presence of both proteins mightprovide increased uptake of iron and protection againstnegative effects of mutation in one of the encoding genesBetween these two proteins hgbA gene was found to beregularly distributed (gt95 prevalence) among isolates [12ndash15 17 18] In the present study 735 of porcine isolates werefound to carry this gene which is lower in comparison toprevious findings that is nearly 100 prevalence (Table 4)[12ndash15] The frequency of hgbB gene varies among strains ofdifferent host origin and also with disease status of the animal[12 15 16 18 19] In this study 722 of the isolates werefound to carry this gene with highest frequency observedamong avian strains (90) which is in agreement withprevious reports (Table 4) [12 17]

Among the genes encoding proteins involved in bacterialcolonization and adhesion ptfA gene has the highest (981)prevalence (Table 3) This gene encodes type 4 fimbriasubunit and has been associated with bovine diseases [19]Worldwide this gene is regularly distributed with more than85 prevalence among P multocida isolates irrespective ofhost origin and capsule type (Table 4)

pfhA gene encoding filamentous haemagglutinin is animportant epidemiological marker and the presence of thisgene has been correlated with occurrence of disease in cattleswine and sheep [12 13 18 19 28] Almost all previous studiesreported low prevalence of this gene with varying frequenciesin between 46ndash52 45ndash60 and 15ndash405 among isolates

of cattle poultry and pig origin respectively (Table 4) [12ndash15] But interestingly very high prevalence 853 (pig) to100 (avian) of this gene was observed among Indianisolates (Table 3)This suggests pfhA genemight be providingsurvival advantage to the bacterium in the host and theoccurrence of horizontal gene transfer has led to such highprevalence among Indian strainsclones

Sialidases play an important role in colonization toepithelial surface They enhance bacterial virulence byunmasking key host receptor and by reducing the effective-ness of mucin [5 29] Of the two sialidases (NanB andNanH)present in P multocida isolates the nanB gene was found inalmost all isolates whereas the prevalence of nanH variedaccording to host origin and geographical location In thisstudy the frequency of nanH gene among poultry isolateswasfound to be low (633) which is in contrast to the reportof Furian et al [17] (Table 4) Similarly the carriage of nanHgene among isolates of pig origin from Indiawas also found tobe lower (676) in comparison to isolates from other parts ofthe globe which reported higher (gt97) frequency (Table 4)[12ndash15]

Dermonecrotoxin (sometimes called P multocida toxin)is encoded by toxA gene This gene was initially detectedin serotype D isolates and was found to be associatedwith atrophic rhinitis in pigs Later on it was detected instrains of serotype A from pigs and other hosts [30] Inthis study toxA gene was detected in 441 of pig isolatesFurther one buffalo and three duck isolates were also foundpositive for toxA gene (Table 3) Two serotype B isolates werefound to carry this gene which is in agreement with ourprevious findings (Sarangi et al submitted for publication)A lysogenic bacteriophage infection of P multocida resultingin horizontal gene transfer could be the reason [31]

The association of virulence associated genes with partic-ular capsular type and host origin was assessed by the Chi-square and Fisherrsquos exact test Out of the 8 virulence associ-ated genes studied toxA pfhA nanB and nanH were found tobe associated (positive or negative) with capsular type pfhAnanB and nanH genes were found to be regularly distributedamong all serotypes with the exception of serotype DNegative association of pfhA gene with CapD strains has beenreported previously [12 14 18] Dermonecrotoxin encoded bytoxA gene was found to be positively associated with capDand CapA Ewers et al [12] observed clear association oftoxA gene with CapD strains which was later supported bysimilar reports from other workers [13 14] Among cattle


Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47


isolates a significant difference (119875 = 0021) was observedin the distribution of hgbB gene among serotypes (Table 3)Similarly for pig isolates the frequency of pfhA and nanHgene among serotypes was found to be statistically significant(Table 3) However as the number of strains tested undereach serotype was less more number of samples should betested before reaching any definite conclusion In order toascertain any trend in the distribution of virulence genesover time period the strains used in the study were dividedinto two groups contemporary (2009ndash2014) and archived(1992ndash2008) and statistical analysis was carried out Butno statistically significant difference was observed betweenthe two groups with respect to virulence gene distribution(Table 1)

The prevalence of virulence associated genes was found tovary among P multocida isolates recovered from various hostspecies Significant association between toxA and nanH geneswith host origin was also observed Dermonecrotoxin genewas found to be positively associated with porcine isolateswhereasnanH genewas found to be positively associatedwithlarge ruminant isolates more specifically with cattle isolateswhich agrees well with the findings of Ewers et al [12]

The combination of genes among P multocida isolateswas assessed by the Chi-square and Fisherrsquos exact testSignificant association was observed between tbpA-hgbAtbpA-hgbB tbpA-pfhA tbpA-nanB tbpA-nanH hgbA-hgbBhgbA-pfhA hgbA-nanB hgbA-nanH pfhA-nanB and pfhA-nanH Similar association among iron acquisition genes aswell as between various virulence associated genes has beenreported previously by Ewers et al [12]

To sum up the present study revealed some uniqueepidemiological information on the prevalence of virulenceassociated genes among Indian strains in comparison to itsequivalents in other parts of the globe The result showsthat with the exception of toxA gene the virulence associatedgenes are regularly distributed among P multocida isolatesThe occurrence of ptfA hgbA and nanH genes among swineisolates of Indian origin was found to be less in comparisonto other countries Gene encoding dermonecrotoxin wasobserved in 176 of the total isolates studied This gene ispresent mostly among swine isolates with few occurrences inbuffalo and duck isolates Interestingly very high prevalenceof tbpA gene was observed among poultry swine and rabbitisolates Likewise very high prevalence of pfhA gene wasobserved among Indian isolates irrespective of host speciesorigin As proper history of majority of the isolates withrespect to its disease status was not available it was notpossible to perform association study between virulencegene and disease status of the animal which could haveenhanced the significance of this study Therefore morenumber of isolates with proper history on disease statusof the host should be carried out in future which willbe helpful to make a more definite conclusion to pro-vide insight into mechanism of pathogenesis association ofgenes with outcome of the disease and in future vaccinestrategies

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

Authors are thankful to Indian Council of AgriculturalResearch (ICAR) New Delhi for providing financial supportunder ldquoAll India Network Programme on HaemorrhagicSepticaemiardquo and the Director Indian Veterinary ResearchInstitute (IVRI) Izatnagar for providing facilities to conductthis study The authors sincerely thank all the scientistsand staff involved in AINP-HS project at IVRI and othercollaborating centres who have been instrumental in isolatingor maintaining the Pasteurella multocida isolates

References

[1] M Harper J D Boyce and B Adler ldquoPasteurella multocidapathogenesis 125 years after pasteurrdquo FEMS Microbiology Let-ters vol 265 no 1 pp 1ndash10 2006

[2] F Dziva A P Muhairwa M Bisgaard and H ChristensenldquoDiagnostic and typing options for investigating diseases asso-ciated with Pasteurella multocidardquo Veterinary Microbiology vol128 no 1-2 pp 1ndash22 2008

[3] S B Shivachandra K N Viswas and A A Kumar ldquoA reviewof hemorrhagic septicemia in cattle and buffalordquoAnimal HealthResearch Reviews vol 12 no 1 pp 67ndash82 2011

[4] R Verma and T N Jaiswal ldquoHaemorrhagic septicaemia vac-cinesrdquo Vaccine vol 16 no 11-12 pp 1184ndash1192 1998

[5] T Hatfaludi K Al-Hasani J D Boyce and B Adler ldquoOutermembrane proteins of Pasteurella multocidardquoVeterinaryMicro-biology vol 144 no 1-2 pp 1ndash17 2010

[6] J Lin S Huang and Q Zhang ldquoOuter membrane proteins keyplayers for bacterial adaptation in host nichesrdquo Microbes andInfection vol 4 no 3 pp 325ndash331 2002

[7] R LDavies RMacCorquodale and S Reilly ldquoCharacterisationof bovine strains of Pasteurella multocida and comparisonwith isolates of avian ovine and porcine originrdquo VeterinaryMicrobiology vol 99 no 2 pp 145ndash158 2004

[8] A W Confer ldquoImmunogens of Pasteurellardquo Veterinary Micro-biology vol 37 no 3-4 pp 353ndash368 1993

[9] A W Confer S H Nutt S M Dabo R J Panciera and GL Murphy ldquoAntibody responses of cattle to outer membraneproteins of Pasteurella multocida A3rdquoThe American Journal ofVeterinary Research vol 57 no 10 pp 1453ndash1457 1996

[10] S M Dabo AW Confer and G L Murphy ldquoOuter membraneproteins of bovine Pasteurella multocida serogroup A isolatesrdquoVeterinary Microbiology vol 54 no 2 pp 167ndash183 1997

[11] V Blume I Luque A I Vela et al ldquoGenetic and virulence-phenotype characterization of serotypes 2 and 9 of Streptococcussuis swine isolatesrdquo International Microbiology vol 12 no 3 pp161ndash166 2009

[12] C Ewers A Lubke-Becker A Bethe S Kiebling M Filterand L H Wieler ldquoVirulence genotype of Pasteurella multocidastrains isolated from different hosts with various disease statusrdquoVeterinary Microbiology vol 114 no 3-4 pp 304ndash317 2006


[13] A Bethe L H Wieler H-J Selbitz and C Ewers ldquoGeneticdiversity of porcine Pasteurella multocida strains from the res-piratory tract of healthy and diseased swinerdquo Veterinary Micro-biology vol 139 no 1-2 pp 97ndash105 2009

[14] X Tang Z Zhao JHu et al ldquoIsolation antimicrobial resistanceand virulence genes of Pasteurella multocida strains from swineinChinardquo Journal of ClinicalMicrobiology vol 47 no 4 pp 951ndash958 2009

[15] N Garcıa J F Fernandez-Garayzabal J Goyache LDomınguez and A I Vela ldquoAssociations between biovar andvirulence factor genes in Pasteurella multocida isolates frompigs in Spainrdquo Veterinary Record vol 169 no 14 p 362 2011

[16] T S P Ferreira M R Felizardo D D Sena de Gobbi etal ldquoVirulence genes and antimicrobial resistance profiles ofPasteurellamultocida strains isolated from rabbits in BrazilrdquoTheScientific World Journal vol 2012 Article ID 685028 6 pages2012

[17] T Q Furian K A Borges S L S Rocha et al ldquoDetectionof virulence-associated genes of Pasteurella multocida isolatedfrom cases of fowl cholera by multiplex-PCRrdquo Pesquisa Vet-erinaria Brasileira vol 33 no 2 pp 177ndash182 2013

[18] K Katsuda K Hoshinoo Y Ueno M Kohmoto and OMikami ldquoVirulence genes and antimicrobial susceptibility inPasteurella multocida isolates from calvesrdquo Veterinary Microbi-ology vol 167 no 3-4 pp 737ndash741 2013

[19] S Verma M Sharma S Katoch et al ldquoProfiling of virulenceassociated genes of Pasteurella multocida isolated from cattlerdquoVeterinary Research Communications vol 37 no 1 pp 83ndash892013

[20] S T Cowan and K J Steel Manual for the Identification ofMedical Bacteria Cambridge University Press Cambridge UK1970

[21] K Wilson ldquoPreparation of genomic DNA from bacteriardquo inCurrent Protocols in Molecular Biology F M Ausubel RBrent R E Kingston et al Eds vol 1 pp 241ndash245 Wiley-Interscience Brooklyn NY USA 1987

[22] KM Townsend A J Frost CW Lee J M Papadimitriou andH J S Dawkins ldquoDevelopment of PCR assays for species- andtype-specific identification of Pasteurella multocida isolatesrdquoJournal of Clinical Microbiology vol 36 no 4 pp 1096ndash11001998

[23] K M Townsend J D Boyce J Y Chung A J Frost and BAdler ldquoGenetic organization of Pasteurella multocida cap lociand development of a multiplex capsular PCR typing systemrdquoJournal of ClinicalMicrobiology vol 39 no 3 pp 924ndash929 2001

[24] S B Shivachandra A A Kumar R Gautam et al ldquoCharacter-ization of avian strains of Pasteurella multocida by restrictionendonuclease and amplified fragment length polymorphismrdquoResearch in Veterinary Science vol 81 no 1 pp 8ndash18 2006

[25] L N Sarangi P Thomas S K Gupta S Kumar K NViswas and V P Singh ldquoMolecular epidemiology of Pasteurellamultocida circulating in India by Multilocus sequence typingrdquoTransboundry and Emerging Diseases 2014

[26] A J Cox M L Hunt J D Boyce and B Adler ldquoFunctionalcharacterization of HgbB a new hemoglobin binding proteinof Pasteurella multocidardquoMicrobial Pathogenesis vol 34 no 6pp 287ndash296 2003

[27] D J Morton PWWhitby H Jin Z Ren and T L Stull ldquoEffectof multiple mutations in the hemoglobin- and hemoglobin-haptoglobin-binding proteins HgpA HgpB and HgpC ofHaemophilus influenzae type brdquo Infection and Immunity vol 67no 6 pp 2729ndash2739 1999

[28] J Shayegh S Atashpaz and M S Hejazi ldquoVirulence genesprofile and typing of ovine Pasteureila multocidardquoAsian Journalof Animal and Veterinary Advances vol 3 no 4 pp 206ndash2132008

[29] E Vimr and C Lichtensteiger ldquoTo sialylate or not to sialylatethat is the questionrdquo Trends in Microbiology vol 10 no 6 pp254ndash257 2002

[30] Z Jaglic Z Kucerova K Nedbalcova I Pavlik P Alexa andM Bartos ldquoCharacterisation and comparison of Pasteurellamultocida isolated fromdifferent species in the Czech Republiccapsular PCR typing ribotyping and dermonecrotoxin produc-tionrdquo Veterinarni Medicina vol 50 no 8 pp 345ndash354 2005

[31] G D Pullinger T Bevir and A J Lax ldquoThe Pasteurellamultocida toxin is encoded within a lysogenic bacteriophagerdquoMolecular Microbiology vol 51 no 1 pp 255ndash269 2004

[32] J A Ogunnariwo and A B Schryvers ldquoCharacterization ofa novel transferrin receptor in bovine strains of Pasteurellamultocidardquo Journal of Bacteriology vol 183 no 3 pp 890ndash8962001

[33] A Jain P Thomas K N Viswas V P Singh and S K GuptaldquoCloning and sequence analysis of outer membrane proteingenes of Pasteurella multocida serotype B2rdquo Indian Journal ofComparative Microbiology Immunology Infectious Diseases vol34 no 1 pp 24ndash28 2013

[34] C A Lichtensteiger S M Steenbergen R M Lee D D Polsonand E R Vimr ldquoDirect PCR analysis for toxigenic Pasteurellamultocidardquo Journal of Clinical Microbiology vol 34 no 12 pp3035ndash3039 1996

Submit your manuscripts athttpwwwhindawicom

Veterinary MedicineJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Veterinary Medicine International



International Journal of

Microbiology


AnimalsJournal of

EcologyInternational Journal of


PsycheHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Evolutionary BiologyInternational Journal of


Hindawi Publishing Corporationhttpwwwhindawicom

Applied ampEnvironmentalSoil Science

Volume 2014

Biotechnology Research International


Agronomy




Journal of Parasitology Research

Hindawi Publishing Corporation httpwwwhindawicom


Volume 2014

Zoology

GenomicsInternational Journal of


InsectsJournal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


VirusesJournal of

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Cell BiologyInternational Journal of



Case Reports in Veterinary Medicine


The previous study carried out in our laboratory on car-riage of 19 virulence genes among Pmultocida isolates recov-ered from small ruminants revealed some novel informationon the frequency of virulence genes like very high prevalenceof pfhA gene 489 prevalence of toxA gene with the highestprevalence among serotype A followed by serotype D andin one isolate each of capsular types B and F (Sarangi etal submitted for publication) These findings from smallruminant isolates encouraged sampling of more isolates ofIndian origin from various hosts to have a clear under-standing on the heterogeneity of the bacteria Therefore thisstudy was extended to P multocida isolates recovered frommultiple host species for 8 important virulence associatedgenes encoding proteins involved in bacterial survival andpathogenesis It included genes encoding transferrin bindingprotein (TbpA) and haemoglobin binding protein (HgbAHgbB) associated with iron acquisition filamentous haemag-glutinin (PfhA) subunit of type IV fimbriae (PtfA) sialidases(NanB NanH) involved in initial colonization and adhesionand dermonecrotoxin (ToxA)

2 Materials and Methods

21 Bacterial Strains In the present study 108 P multocidaisolates recovered from large ruminants (buffalo 119899 = 23cattle 119899 = 18) avians (chicken 119899 = 18 duck 119899 = 8 andemu 119899 = 4) swine (119899 = 34) and rabbit (119899 = 3) maintainedat Division of Bacteriology amp Mycology Indian VeterinaryResearch Institute Izatnagar were used Selections of isolateswere carried out on the basis of host origin year andplace of isolation in order to incorporate isolates from allover India The details of the isolates (isolate number hostorigin capsular type year and place of isolation and diseasesymptom (if available)) are given in Table 1

22 Confirmation of P multocida Isolates The isolates wererevived in brain heart infusion broth by 18ndash24 h incubationat 37∘C and plated subsequently onto blood agar to study cul-tural characteristics The cultures were then tested for purityby biochemical tests as per standard techniques [20] Thegenomic DNA of the isolates was extracted by CTABmethod[21] and the isolates were reconfirmed as Pmultocida by PM-PCR followed by determination of capsular type bymultiplexPCR [22 23]

23 Detection of Virulence Associated Genes by PCR Theisolates were then subjected to screening of 8 virulence genesencoding iron binding proteins (TbpA HgbA HgbB) colo-nization and adhesion related protein (PfhA PtfA) sialidases(NanB NanH) and dermonecrotoxin (ToxA) by individualPCR reactions utilizing oligonucleotide primers describedpreviously The details of the virulence genes sequences ofthe oligonucleotide primers and citations used are listed inTable 2

24 Statistical Analysis Statistical analysis of the data gener-ated from the study was performed with SPSS 160 (SPSS IncChicago) 119875 values of lt005 were considered as statisticallysignificant

3 Results and Discussion

P multocida is an economically important veterinarypathogen causing wide range of diseases in livestock andpoultry The bacteria have been classified into five capsulartypes (A B D E and F) based on capsular typing witheach capsule type being predominantly associated with aparticular disease in a host species But isolation of othercapsular types from such hosts by cross species infection isnot uncommon [2 7] Ability of the bacteria to infect andsurvive in several hosts as commensal exposes it to variousselection pressures resulting in emergence of divergentstrains in field scenario Molecular epidemiological studyby employing REP-PCR ERIC-PCR MLST analysis and soforth has confirmed the diversity of P multocida circulatingin India and also the possibility of transboundary spreadof strains across evolutionary time [24 25] Therefore adetailed study on the presence of virulence associated genesrecovered from different host species in Indian subcontinentwill be helpful to understand the disease process and todevelop disease control measures in future

In this study P multocida isolates recovered from varioushost species were screened for presence of 8 importantvirulence associated genes (tbpA hgbA hgbB pfhA ptfAnanB nanH and toxA) involved in bacterial pathogenesisThe results confirmed that with the exception of toxA geneall other virulence associated genes are regularly distributedamong the isolates of different host origin The result ofindividual PCR reaction for each isolate is presented inTable 1 Among the genes encoding iron binding proteinstbpA gene was present in 824 of isolates which range from696 in buffalo to 100 in cattle emu and rabbits (Table 3)Similarly hgbA gene was found to be regularly distributedamong all isolates affecting different hosts with the lowestprevalence among swine isolates (735) Gene hgbB hasthe lowest prevalence among the three iron binding proteinsscreened in this study and was found in 722 of the isolatesThe percentage prevalence of this gene was found to be moreamong avian isolates (90) in comparison to large ruminants(659) and swine (676) isolates Of the two sialidasespresent in P multocida isolates the percentage prevalence ofnanB gene was found to be more than nanH gene Overallvery high prevalence of pfhA gene (935) was observedin this study with 100 prevalence among avian isolatesDermonecrotoxin gene (toxA) was found only in 176 ofstrains with majority of the isolates belonging to porcineorigin One buffalo and 3 duck isolates were also found tocarry toxA gene (Table 3)

Iron acquisition and uptake are essential for bacterialsurvival and many bacteria have developed different ironsequestering system for uptake of ironThe expression of ironacquisition proteins increases under iron limiting conditionas well as in vivo condition (reviewed in [5]) P multocidautilizes various receptors for adapting to variations in supplyof different haem iron sources [5] Among these TbpAprotein is necessary for extraction of iron from transferrinand has been reported to be an important virulence factorand epidemiological marker in cattle [12 19 26] Previousstudies reported that tbpA gene is either absent or rarely





Table 1 Continued



P P P P P A P A



Table 1 Continued




KMT1 PMPCR-FPMPCR-R




bcbD capB FcapB R


dcbF capD FcapD R


ecbJ capE FcapE R


fcbD capF FcapF R



tbpA tbpA FtbpA R


hgbA hgbA FhgbA R


hgbB HgbB FHgbB R


Adhesins

ptfA ptfA FptfA R


pfhA pfhA FpfhA R


Sialidases

NanB NanB FNanB R


nanH nanH FnanH R


Dermonecrotoxin

ToxA ToxA FToxA R





hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()



Table 3 Continued


hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()











Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47








Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of










Table 1 Continued



P P P P P A P A



Table 1 Continued




KMT1 PMPCR-FPMPCR-R




bcbD capB FcapB R


dcbF capD FcapD R


ecbJ capE FcapE R


fcbD capF FcapF R



tbpA tbpA FtbpA R


hgbA hgbA FhgbA R


hgbB HgbB FHgbB R


Adhesins

ptfA ptfA FptfA R


pfhA pfhA FpfhA R


Sialidases

NanB NanB FNanB R


nanH nanH FnanH R


Dermonecrotoxin

ToxA ToxA FToxA R





hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()



Table 3 Continued


hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()











Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47








Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of







Table 1 Continued




KMT1 PMPCR-FPMPCR-R




bcbD capB FcapB R


dcbF capD FcapD R


ecbJ capE FcapE R


fcbD capF FcapF R



tbpA tbpA FtbpA R


hgbA hgbA FhgbA R


hgbB HgbB FHgbB R


Adhesins

ptfA ptfA FptfA R


pfhA pfhA FpfhA R


Sialidases

NanB NanB FNanB R


nanH nanH FnanH R


Dermonecrotoxin

ToxA ToxA FToxA R





hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()



Table 3 Continued


hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()











Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47








Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of









hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()



Table 3 Continued


hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()











Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47








Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of







Table 3 Continued


hgbA()

hgbB()

pfhA()

ptfA()

nanB()

nanH()

toxA()











Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47








Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of







Table4Com

paris

onof

thed

istrib

utionof

virulencea

ssociatedgenesa

mon

gPa

steurellamultocid

aiso

latesrecovered

from

vario

usho

stspeciesa

crossthe

glob

e

Geneho

st

Cattle

Buffa

loPo

ultry

Pig

Rabb

it

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

Cou

ntry

(reference)

Noof

strains

teste

d

Prevalence

()

tbpA

India(

thisstu

dy)

18100

India(

thisstu

dy)

23696

India(

thisstu

dy)

30833

India(

thisstu

dy)

34794

India(

thisstu

dy)

03100

Germany[12]

104

702

Germany[12]

07571

Germany[12]

2000

Germany[12]

5200

Germany[12]

2000

Japan[18]

378

762

Spain[15]

205

00

Brazil[16]

4686

India[

19]

23100

Germany[13]

382

00

hgbA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23826

India(

thisstu

dy)

30867

India(

thisstu

dy)

34735

India(

thisstu

dy)

03100

Germany[12]

104

952

Germany[12]

07100

Germany[12]

20900

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

955

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46739

India[

19]

23100

Germany[13]

382

100

China[

14]

233

966

hgbB

India(

thisstu

dy)

1866

7India(

thisstu

dy)

23652

India(

thisstu

dy)

30900

India(

thisstu

dy)

34676

India(

thisstu

dy)

03333

Germany[12]

104

577

Germany[12]

07857

Germany[12]

20850

Germany[12]

52865

Germany[12]

20100

Japan[18]

378

614

Brazil[17]

25100

Spain[15]

205

605

Brazil[16]

46304

India[

19]

23261

Germany[13]

382

843

ptfA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34100

India(

thisstu

dy)

03100

Germany[12]

104

990

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

947

Brazil[17]

25920

Spain[15]

205

100

Brazil[16]

46934

India[

19]

23869

Germany[13]

382

100

China[

14]

233

936

pfhA

India(

thisstu

dy)

18944

India(

thisstu

dy)

23957

India(

thisstu

dy)

30100

India(

thisstu

dy)

34853

India(

thisstu

dy)

03100

Germany[12]

104

462

Germany[12]

07100

Germany[12]

20450

Germany[12]

52212

Germany[12]

20750

Japan[18]

378

524

Brazil[17]

25600

Spain[15]

205

405

Brazil[16]

4600

India[

19]

23100

Germany[13]

382

209

China[

14]

233

150

nanB

India(

thisstu

dy)

18100

India(

thisstu

dy)

23100

India(

thisstu

dy)

30100

India(

thisstu

dy)

34882

India(

thisstu

dy)

03100

Germany[12]

104

100

Germany[12]

07100

Germany[12]

20100

Germany[12]

52100

Germany[12]

20100

Japan[18]

378

100

Brazil[17]

25100

Spain[15]

205

100

Brazil[16]

46956

India[

19]

2300

Germany[13]

382

100

China[

14]

233

815

nanH

India(

thisstu

dy)

18100

India(

thisstu

dy)

23783

India(

thisstu

dy)

30633

India(

thisstu

dy)

34676

India(

thisstu

dy)

03100

Germany[12]

104

885

Germany[12]

07100

Germany[12]

20650

Germany[12]

52981

Germany[12]

20100

Japan[18]

378

884

Brazil[17]

25960

Spain[15]

205

100

Brazil[16]

46673

India[

19]

23100

Germany[13]

382

100

China[

14]

233

970

toxA

India(

thisstu

dy)

1800

India(

thisstu

dy)

2343

India(

thisstu

dy)

30100

India(

thisstu

dy)

3444

1India(

thisstu

dy)

0300

Germany[12]

104

58

Germany[12]

0700

Germany[12]

2050

Germany[12]

52365

Germany[12]

2000

India[

19]

2300

Brazil[17]

2500

Spain[15]

205

78Brazil[16]

4600

Germany[13]

382

34

China[

14]

233

47








Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of













Acknowledgments


References











































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of




































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of






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