Aims: This study aimed to evaluate the effects of three Bacillus probiotics on Salmonella Typhimurium, and interleukin-8 (IL-8) gene expression in the co-culture of the Bacillus and the pathogen in vitro. Methods and results: Bacillus subtilis, Bacillus indicus and Bacillus coagulans were initially turned to spore and heat-inactivated forms. The cellular damages of the probiotics on the HT-29 cells were investigated individually and in combination with S. Typhimurium using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and fluorescence assays. To extract cell free supernatants (CFS) of the probiotics, they were cultured in selective media. The inhibitory activity of CFSs were then assayed against the pathogen. The gene expression of IL-8 of the HT-29 cells was evaluated by real-time PCR in all the groups. The results showed that the CFSs of three probiotics could inhibit the growth of S. Typhimurium by more than 50%. Inhibitory effects of B. indicus and B. subtilis CFSs were related to the production of pepsin-sensitive compounds, except B. coagulans in which the high inhibitory effect was due to organic acids. The spores of the three probiotics and the heat-inactivated forms of B. subtilis and B. coagulans could reduce the cytotoxicity of S. Typhimurium. The cell viability also increased applying both forms probiotics against the pathogen. In all co-culture groups, the IL-8 gene expression induced by S. Typhimurium was reduced. Conclusions: The three Bacillus probiotics can be considered as proper candidates for the prevention and treatment of S. Typhimurium food poisoning.

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