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Background   Campylobacter fetus  subspecies  venerealis  is the causative agent of bovine genital campylobacteriosis, asymptomatic in bulls the disease is spread to female cattle causing extensive reproductive loss. The microbiological and molecular differentiation of  C. fetus  subsp.  venerealis  from  C. fetus  subsp.  fetus  is extremely difficult. This study describes the analysis of the available  C. fetus  subsp.  venerealis  AZUL-94 strain genome (~75–80%) to identify elements exclusively found in  C. fetus  subsp.  venerealis  strains as potential diagnostic targets and the characterisation of subspecies virulence genes.    Results  Eighty Kb of genomic sequence (22 contigs) was identified as unique to  C. fetus  subsp.  venerealis  AZUL-94 and consisted of type IV secretory pathway components, putative plasmid genes and hypothetical proteins. Of the 9 PCR assays developed to target  C. fetus  subsp.  venerealis  type IV secretion system genes, 4 of these were specific for  C. fetus  subsp.  venerealis  biovar  venerealis  and did not detect  C. fetus  subsp.  venerealis  biovar  intermedius . Two assays were specific for  C. fetus  subsp.  venerealis  AZUL-94 strain, with a further single assay specific for the AZUL-94 strain and  C. fetus  subsp.  venerealis  biovar  intermedius  (and not the remaining  C. fetus  subsp.  venerealis  biovar  venerealis  strains tested).  C. fetus  subsp.  fetus  and  C. fetus  subsp.  venerealis  were found to share most common  Campylobacter  virulence factors such as SAP, chemotaxis, flagellar biosynthesis, 2-component systems and cytolethal distending toxin subunits (A, B, C). We did not however, identify in  C. fetus  the full complement of bacterial adherence candidates commonly found in other  Campylobacter  spp.    Conclusion  The comparison of the available  C. fetus  subsp.  venerealis  genome sequence with the  C. fetus  subsp.  fetus  genome identified 80 kb of unique  C. fetus  subsp.  venerealis  AZUL94 sequence, with subsequent PCR confirmation demonstrating inconsistent amplification of these targets in all other  C. fetus  subsp.  venerealis  strains and biovars tested. The assays developed here highlight the complexity of targeting strain specific virulence genes for field studies for the molecular identification and epidemiology of  C. fetus .

Genomic analysis of Campylobacter fetus subspecies: identification of candidate virulence determinants and diagnostic assay targets