Phylogenetic Analysis of Atypical Hemolysin Gene in Vibrio campbellii and Effects of Cultivation Salinity and pH on Hemolytic Activity and Virulence

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The purposes of this study were to analyze the atypical hemolysin gene of V. campbellii isolates, and to evaluate the effects of cultivation salinity (0.5, 1.5, and 3.0 % NaCl) and pH (5.0, 7.3, and 8.6) on the hemolytic activity and virulence of V. campbellii. Phylogenetic analysis of atypical hemolysin gene sequences obtained from V. campbellii demonstrated 84 - 85 % identity with the hlyA of V. cholerae. V. campbellii grown at 1.5 or 3.0 % NaCl, which exhibited significant higher hemolytic activity compared to those previously grown at 0.5 % NaCl. Maximum hemolytic activity was observed among acid-adapted V. campbellii which was previously grown at pH 5.0 for 5 h. Likewise, its virulence against Galleria mellonella was enhanced (~20 times) in comparison to that of non-adapted V. campbellii. Based on our results, it seems that V. campbellii might have acquired hemolysin gene from V. cholerae. Moreover, both cultivation salinity and pH are deemed important for the hemolytic activity and virulence of V. campbellii. This will be useful for the environmental control of this pathogen in aquaculture.

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