Effects of varying concentrations of sodium chloride and acidic conditions on the behavior of Vibrio parahaemolyticus and Vibrio vulnificus cold-starved in artificial sea water microcosms

There has been limited information available on the behavior of Vibrio parahaemolyticus and Vibrio vulnificus as a function of higher levels of NaCl in combination with acidic pH. In the present study, bacterial suspensions were transferred into artificial seawater (pH 4–7) microcosms containing 0.75% NaCl and supplemented with 5, 10, and 30% NaCl, respectively. Each of V. parahaemolyticus and V. vulnificus was inoculated in these microcosms and fermented seafood, and then stored at 4 °C until the microbial populations reached below the detectable levels on agar plates (thiosulphate-citrate-bile salts-sucrose agar and tryptic soy agar amended with 3% NaCl). Consequently, V. parahaemolyticus ATCC 27969, V. parahaemolyticus ATCC 33844, and V. vulnificus ATCC 33815 rapidly reached the viable-but-nonculturable (VBNC) state with increasing levels (≤30%) of NaCl at 4 °C. Within seven days, these pathogens in seafood appeared to enter the VBNC state at 4 °C, as shown by the fluorescence microscopic assay.