Abstract: The inhibitory effects of Lactobacillaceae L1 and its metabolic product on pathogen Vibrio sp.T1, T2 were examined by a zone plate inhibitory method. The results show that the antibacterial activity in supernatant liquid of Lactobacillaceae to T1 and T2 is similar to that of fermentation liqud, but the inhibitory effect of L1 on Vibrio T1 is stronger than that on T2. Fermentation liquid has a higher antibacterial activity than supernatant liquid, indicates that Lactobacillaceae and its metabolic product have a conjunct inhibitory effect on Vibrio T1 and T2. The antibacterial activity of fermentation liquid still remains after 3 times dilution. The antibacterial activity of metabolic product of lactobacillaceae to Vibrio T1 and T2 vary from different growth periods, with being stronger in the decline period stability period growth period. The inhibitory activity of fermentation liquid of L1 treated with boiled water for 15 min is invariable, shows that temperature tolerance of the inhibitory substance is better.