Multilocus Sequencing of Corynebacterium pseudotuberculosis Biotype Ovis Strains.

Boglárka Sellyei,Dániel Bartha,László Makrai,István Hajtós,László Fodor,Krisztián Bányai

doi:10.1155/2017/1762162

Abstract

Thirteen Corynebacterium pseudotuberculosis biotype ovis strains isolated from clinical cases of caseous lymphadenitis in Hungary were characterised using multilocus sequencing and their phylogenetic comparison was carried out on the basis of four housekeeping genes (groEL1, infB, dnaK, and leuA). The in silico analysis of the 16 frequently studied housekeeping genes showed that C. pseudotuberculosis strains could be readily distinguished from C. diphtheriae and C. ulcerans strains; however, sequences of the same genes in the two biotypes of the C. pseudotuberculosis were highly similar; the heterogeneity values were low. Genes dnaK, infB, groEL1, and leuA showed marked genetic variation within C. pseudotuberculosis, and strains of the two biotypes of C. pseudotuberculosis could be differentiated. Analysis of the individual genes showed a fairly conservative nature of C. pseudotuberculosis biotype ovis strains. The greatest genetic differentiation was seen in the dnaK and infB genes and concatenations of these two genes were very useful in the genetic separation of the studied strains.

Highlights

Corynebacterium pseudotuberculosis was isolated from a sheep and described by Preisz [1] as a Gram-positive, facultative intracellular pathogen
Two major biotypes can be distinguished based on the ability of reduction of nitrate; the nitrate negative biotype (C. pseudotuberculosis biotype ovis) causes caseous lymphadenitis (CLA) in small ruminants [2,3,4], whereas the nitrate positive biotype (C. pseudotuberculosis biotype equi) is the causative agent of ulcerative lymphangitis in horses, cows, camels, buffaloes, and occasionally humans [5, 6]
C. pseudotuberculosis biotype equi strains showed greater genetic divergence than C. pseudotuberculosis biotype ovis strains when examined with pulsed field gel electrophoresis (PFGE), BOX-PCR, random amplified polymorphic DNA (RAPD), and amplification of DNA surrounding rare restriction sites (ADSRRS) [3, 15, 16]

Summary

Introduction

Corynebacterium pseudotuberculosis was isolated from a sheep and described by Preisz [1] as a Gram-positive, facultative intracellular pathogen. Two major biotypes can be distinguished based on the ability of reduction of nitrate; the nitrate negative biotype (C. pseudotuberculosis biotype ovis) causes caseous lymphadenitis (CLA) in small ruminants [2,3,4], whereas the nitrate positive biotype (C. pseudotuberculosis biotype equi) is the causative agent of ulcerative lymphangitis in horses, cows, camels, buffaloes, and occasionally humans [5, 6]. C. pseudotuberculosis biotype equi strains showed greater genetic divergence than C. pseudotuberculosis biotype ovis strains when examined with pulsed field gel electrophoresis (PFGE), BOX-PCR, random amplified polymorphic DNA (RAPD), and amplification of DNA surrounding rare restriction sites (ADSRRS) [3, 15, 16]

Objectives

Methods

Results

Conclusion