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Background    Vibrio parahaemolyticus  is a recognized human enteropathogen. Thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) as well as the type III secretion system 2 (T3SS2) are considered as major virulence factors. As  tdh  positive strains are not detected in coastal waters of Germany, we focused on the characterization of  trh  positive strains, which were isolated from mussels, seawater and patients in Germany.    Results  Ten  trh  harbouring  V .  parahaemolyticus  strains from Germany were compared to twenty-one  trh  positive strains from other countries. The complete  trh  sequences revealed clustering into three different types:  trh1  and  trh2  genes and a pseudogene  Ψtrh . All German isolates possessed alleles of the  trh2  gene. MLST analysis indicated a close relationship to Norwegian isolates suggesting that these strains belong to the autochthonous microflora of Northern Europe seawaters. Strains carrying the pseudogene  Ψtrh  were negative for T3SS2β effector  vopC . Transcription of  trh  and  vopC  genes was analyzed under different growth conditions.  Trh2  gene expression was not altered by bile while  trh1  genes were inducible.  VopC  could be induced by urea in  trh2  bearing strains. Most  trh1  carrying strains were hemolytic against sheep erythrocytes while all  trh2  positive strains did not show any hemolytic activity. TRH variants were synthesized in a prokaryotic cell-free system and their hemolytic activity was analyzed. TRH1 was active against sheep erythrocytes while TRH2 variants were not active at all.    Conclusion  Our study reveals a high diversity among  trh  positive  V .  parahaemolyticus  strains. The function of TRH2 hemolysins and the role of the pseudogene  Ψtrh  as pathogenicity factors are questionable. To assess the pathogenic potential of  V .  parahaemolyticus  strains a differentiation of  trh  variants and the detection of T3SS2β components like  vopC  would improve the  V .  parahaemolyticus  diagnostics and could lead to a refinement of the risk assessment in food analyses and clinical diagnostics.

Characterization of trh2 Harbouring Vibrio parahaemolyticus Strains Isolated in Germany