Detection and Isolation of Vibrio spp. in Seafood With Cultural and Molecular Method

Vibrio spp. are gram-negative, facultative anaerobic, non-spore-forming bacilli which are oxidase positive and halophilic bacteria widely spread in seaand brackwater worldwide. The genus Vibrio includes several foodborne pathogens which cause a spectrum of clinical conditions including septicaemia, cholera and milder forms of gastroenteritis. The species most commonly associated with foodborne transmission include V. cholerae, V. parahaemolyticus, and V. vulnificus. V. parahaemolyticus causes a much milder diarrhea than cholera. In this study, totally 89 seafood samples were analysed for the detection of Vibrio spp. These samples are: calamary, mackerel, shrimp, red mullet, trout. The approach involved detecting Vibrio spp. by cultural and molecular method. For cultural method involves an enrichment phase with incubation at 37°C for 24 h in alkaline peptone water and an isolation phase on selectivedifferential medium thiosulphate citrate bile salts sucrose (TCBS), Modified cellobiose polymyxin B colistin agar (mCPC) plates incubating at 37°C for 24 h. For the detection and identification of Vibrio spp. with molecular method (V. parahaemolyticus, V. cholerae and V. vulnificus) three probe-based multiplex real-time PCR systems were used. Real Time PCR process were carried out