Molecular Characterization and Antibiotic Susceptibility of Vibrio vulnificus in Retail Shrimps in Hangzhou, People's Republic of China

Abstract

Vibrio vulnificus is a gram-negative bacterium that occurs naturally in estuarine and marine water and is associated with wound infections or septicemia related to the consumption of raw shellfish in humans. The molecular characteristics and antibiotic susceptibilities of V. vulnificus strains in shrimps from retail markets in Hangzhou, People's Republic of China, were investigated in this study. Thirty-three samples were positive for V. vulnificus in 78 shrimp samples which were collected from 15 retail markets between July and August 2012; the most-probable-number values ranged from 3 to 1,600 g−1 in these positive samples, with a median most-probable-number value of 72 g−1. Twenty-five biotype 1 strains and eight biotype 2 strains were identified by biochemical tests, and all strains could be definitively genotyped. By 16S rRNA genotyping, 21.2% (7 of 33) were classified as genotype A, 63.6% (21 of 33) as genotype B, and 15.2% (5 of 33) as genotype AB, while by virulence-correlated gene (vcg) typing, 21.2% (7 of 33) were characterized as genotype E and 78.8% (26 of 33) were genotype C. More than 50% of those isolates were identified as the potentially virulent type vcg type C–16S rRNA B (CB). The antibiotic susceptibilities of the V. vulnificus strains to 21 antimicrobial agents were tested as well. Some strains showed resistance or intermediate resistance to cefepime (3.03%), tetracycline (6.06%), aztreonam (24.24%), streptomycin (45.45%), gentamicin (93.94%), tobramycin (100%), and cefazolin (100%). Multiple-locus variable-number tandem repeat-based fingerprinting analysis (MLVA) was successfully applied to these 33 isolates and yielded 30 patterns that clustered into two MLVA groups; with a calculated Simpson's index of diversity of 0.994, this revealed that MLVA had great discriminating power for V. vulnificus. To minimize the potential risk of V. vulnificus infections due to the consumption of raw shrimp, it is necessary to monitor the hygiene status of seafood.