safety of Bacillus subtilis and Bacillus indicus as food probiotics

• Published in: Journal of applied microbiology Vol. 105; no. 2; pp. 510 - 520
• Main Authors: Hong, H.A, Huang, J.-M, Khaneja, R, Hiep, L.V, Urdaci, M.C, Cutting, S.M
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01-08-2008; Blackwell Publishing Ltd; Blackwell Science
• Subjects: Animals; Bacillus; Bacillus subtilis; Bacillus subtilis - pathogenicity; Bacillus subtilis - physiology; Bacterial Adhesion; Biological and medical sciences; Consumer Product Safety; Enterotoxins - analysis; Enterotoxins - genetics; Food Microbiology; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Guinea Pigs; Humans; Intestines - microbiology; Microbiology; Probiotics; Rabbits; safety; spore formers; Spores, Bacterial; toxicity; Toxicity Tests; Virulence; Probiotic; spore formers; Bacillales; Bacillus subtilis; Applied microbiology; Toxicity; Bacteria; Foodstuff; Safety; Bacillaceae; Spores; probiotics
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2008.03773.x

To conduct in vitro and in vivo assessments of the safety of two species of Bacillus, one of which, Bacillus subtilis, is in current use as a food supplement. Cultured cell lines, Caco-2, HEp-2 and the mucus-producing HT29-16E cell line, were used to evaluate adhesion, invasion and cytotoxicity. The Natto strain of B. subtilis was shown to be able to invade and lyse cells. Neither species was able to adhere significantly to any cell line. The Natto strain was also shown to form biofilms. No strain produced any of the known Bacillus enterotoxins. Disc-diffusion assays using a panel of antibiotics listed by the European Food Safety Authority (EFSA) showed that only Bacillus indicus carried resistance to clindamycin at a level above the minimum inhibitory concentration breakpoints set by the EFSA. In vivo assessments of acute and chronic dosing in guinea pigs and rabbits were made. No toxicity was observed in animals under these conditions. Bacillus indicus and B. subtilis should be considered safe for oral use although the resistance of B. indicus to clindamycin requires further study. The results support the use of B. subtilis and B. indicus strains as food supplements.