Isolation and Identification Species of Vibrio Genus from Fresh and Frozen Shrimp and Confirmation the Identification of Vibrio parahaemolyticus by Polymerase Chain Reaction Technique

Abstract

This research was concerned with the isolation and identification of Vibrio parahaemolyticus from seafood (shrimp) samples. (40) samples of freshly harvested shrimp from the local markets in Al-Basra city and twenty (20) samples of frozen shrimp (Turkey origin) were collected. Two enrichment methods were used for the enhancement of the growth and isolation of vibrio species, which include enrichment in non selective medium Alkaline peptone water (APW) for 8 hours and then in selective broth salt polymyxin broth (SPB) and then plated on two solid selective media, ThioSulphate Citrate Bile Sucrose agar (TCBS) and Arabinose Ammonium Sulphate Cholate agar (AASC) and morphological, cultural and biochemical tests were performed on the colonies grown on selective media, then we confirmed the identification by using the chromogenic medium ChromagarTM Vibrio for the first time locally. Finally tox R based polymerase chain reaction was used to cofirm the identification of V. parahaemolyticus. The results showed the isolation of Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, Vibrio mimicus in percentage 45%, 20%, 37.5%, 17.5% from fresh shrimp, 35%, 20%, 25%, 15% from the frozen shrimp respectively. The results also showed that there is an accordenance between biochemical tests and ChromagarTM Vibrio. The results of tox R based PCR revealed that 22l25 of V. parahaemolyticus contain the tox R gene which is species-specific gene.