Aims: This study was carried out to investigate the occurrence of potentially pathogenic species of Vibrio in French marine and estuarine environments. Methods and Results: Samples of coastal waters and mussels collected between July and September 1999 were analysed by culture, using selective media including thiosulphate-citratebile salts-sucrose and modified cellobiose-polymixin B-colistin agar. Presumptive Vibrio colonies were isolated and identified using selected biochemical tests. Specific primers based on flanking sequences of the cytolysin, vvhA gene, pR72H DNA fragment and 16S-23S rRNA intergenic spacer region (ISR) were used in a polymerase chain reaction (PCR) to confirm the identification of Vibrio vulnificus, V. parahaemolyticus and V. cholerae, respectively. In this study, V. alginolyticus (99 of 189) was the predominant species, followed by V. parahaemolyticus (41 of 189), V. vulnificus (20 of 189) and non-O1/non-O139 V. cholerae (three of 189). All 20 V. vulnificus isolates showed PCR amplification of the vvhA gene, 16 of which had been isolated from estuarine water. The PCR amplification of the pR72H DNA fragment in 41 V. parahaemolyticus isolates generated two unique amplicons of 387 and 320 bp. The latter, present in 24AE4% of these isolates, had not previously been found in V. parahaemolyticus strains examined to date. Amplification of the trh gene in two of the isolates suggested these to be virulent strains. Three strains identified as V. cholerae by amplification of the 16S-23S rRNA ISR were confirmed to be non-cholera (non-O1/non-O139) strains.
Conclusions:The results of this study demonstrated the presence of pathogenic Vibrio species in French coastal waters. Furthermore, the PCR approach proved useful for the rapid and reliable confirmation of species identification. Significance and Impact of the Study: These findings indicate the potential sanitary risk associated with the presence of pathogenic Vibrio spp. in cultivated mussels and in the aquatic environment. The PCR can be used to detect pathogenic vibrios directly in environmental samples.
A significant proportion of shrimps marketed and consumed in Morocco are caught in the coastal region of the city of Agadir. This study provides interesting data of prevalence of Vibrio spp. in raw shrimps as well as better understanding of their potential virulence. It is apparent from this study that genes and primers used in multiplex PCR for identification and detection of virulence factors, can be used to monitor shrimps for the presence of potentially pathogenic strains of Vibrio cholerae and Vibrio parahaemolyticus. The results highlight the added value of using a chromogenic medium for research and isolation of pathogenic Vibrio in seafood, more specific and accurate than TCBS.
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