Virulence Gene Pattern of Pasteurella multocida Isolates of Buffalo in Association to Capsule Biosynthesis Genes

Abstract

Background: Pasteurella multocida is the causative agent of many economically important diseases in a wide range of hosts. The mechanisms by which these bacteria can invade the mucosa, evade innate immunity and cause systemic disease are slowly being elucidated. Many key virulence factors are yet to be identified, including those required for initial attachment and invasion of host cells and for persistence in a relatively nutrient poor and hostile environment. This has led to intensive research to understand host adaptation mechanisms and virulence factors in order to develop effective vaccines. Methods: The present study was carried out to know the distribution of virulence genes viz., haemoglobin binding proteins (hgbA and hgbB), outer membrane protein (ompH), fimbrial antigen (ptfA), filamentous haemagglutinin (pfhA) and transferrin binding protein (tbpA) by PCR in P. multocida CapA isolates from apparently healthy or carrier animals and CapB isolates from field Haemorrhagic septicemia (HS) cases to monitor the epidemiological associations of virulence genes in Cap A and Cap B isolates. Result: The study revealed that all the six virulence associated genes were present in Cap B isolates. None of the Cap A isolates harboured tbpA and pfhA genes. These two genes were closely related to serotype B causing Haemorrhagic septicemia and were epidemiologically associated with disease status.

Molecular detection and genotyping of P. multocida, M. hemolytica, and B. trehalosi isolates targeting the virulence associated genes from Ethiopian cattle and sheep

“…For most PCR reactions, 5µl template DNA was added to the total 20 µl reaction mixture containing 4.5µl of nucleasefree water, 2µl of each forward and reverse primer pair (5pmol/µl), 5 µl of 10x DreamTaq™ Buffer that contains 20mM MgCl 2 , 5µl of dNTP Mix (2mM each) and 1.5µl of Dream Taq™ polymerase Ampli cation protocols for the capsular biosynthesis gene of P. multocida, PHSSA-1, and methylene transferase encoding gene (Rpt2) of M. hemolytica and manganese-dependent superoxide dismutase encoding gene of B. trehalosi (BtsodA) were used as previously described by Townsend et al [33], Kumar et al [28] and Ewers et al [34], respectively. PCR thermal cycling conditions for the iron acquisition TbpA2 and FbpA genes were also prepared as previously stated by Sujatha et al [35] and Akalu [36], respectively. PCR instructions provided by the University of Calgary were used as a PCR protocol for surface lipoproteins (PmSLP) locus [37].…”

Molecular Detection and Genotyping of P. Multocida, M. Hemolytica, and B. Trehalosi Isolates Targeting the Virulence Associated Genes from Ethiopian Cattle and Sheep

“…For most PCR reactions, 5µl template DNA was added to the total 20 µl reaction mixture containing 4.5µl of nuclease-free water, 2µl of each forward and reverse primer pair (5pmol/µl), 5 µl of 10x DreamTaq™ Buffer that contains 20mM MgCl2, 5µl of dNTP Mix (2mM each) and 1.5µl of Dream Taq™ polymerase Amplification protocols for the capsular biosynthesis gene of P. multocida, PHSSA-1, and methylene transferase encoding gene(Rpt2) of M. hemolytica and manganese-dependent superoxide dismutase encoding gene of B. trehalosi (BtsodA) were used as previously described by respectively [29, 33 and 34]. PCR thermal cycling conditions for the iron acquisition TbpA2 and FbpA genes were also prepared as previously stated by respectively [35,36]. PCR instructions provided by the University of Toronto were used as a PCR protocol for surface lipoproteins (PmSLP) locus [37].…”