Characteristics of Vibrio parahaemolyticus isolates obtained from crayfish (Procambarus clarkii) in freshwater

•This study sheds light on the relationship between V. parahaemolyticus and crayfish from freshwater.•The genotypes and the evolutionary relationship between the isolates and pandemic strains were identified.•Among all 96 isolates analyzed, 53 sequence types (STs) were identified, 38 of which were novel STs.•Forty isolates contained more than two genes with possible roles in pathogenicity.•Zebrafish and mouse infection models revealed the potential virulence of V. parahaemolyticus in freshwater.

Vibrio parahaemolyticus usually occurs in coastal areas and is generally recognized as a marine bacterium. It has become the leading cause of gastroenteritis worldwide. In the present study, 96 V. parahaemolyticus isolates were obtained from freshwater crayfish (Procambarus clarkii) and classified by multilocus sequence typing. Fifty-three sequence types (STs) were identified among the 96 isolates analyzed, 38 of which were novel STs. These isolates fell into six groups and 42 singletons, suggesting a high level of genetic diversity. Screening for 9 virulence and virulence-related genes in the isolates revealed that 40 isolates contained more than two genes with possible roles in pathogenicity. The virulence of the representative isolates VP66 (trh+, ureC+, T3SS1+, T3SS2β+, T6SS2+) and VP80 (T3SS1+, T6SS1+, T6SS2+) were further assessed in zebrafish and mouse infection model in vivo, and the tested isolates were shown to be lethal to both zebrafish and mice. These results suggest that crayfish may serve as a carrier of V. parahaemolyticus in freshwater, and that some isolates may have the potential to cause foodborne disease in humans.