Inhibition of the cytotoxic effect of Clostridium difficile in vitro by Clostridium butyricum MIYAIRI 588 strain

• Published in: Journal of medical microbiology Vol. 60; no. 11; pp. 1617 - 1625
• Main Authors: WOO, Timothy D. H, OKA, Kentaro, TAKAHASHI, Motomichi, HOJO, Fuhito, OSAKI, Takako, HANAWA, Tomoko, KURATA, Satoshi, YONEZAWA, Hideo, KAMIYA, Shigeru
• Format: Journal Article
• Language: English
• Published: Reading Society for General Microbiology 01.11.2011
• Subjects: Animals; Antibiotics; Bacteriology; Biological and medical sciences; Blotting, Western; Cell culture; Cell Survival - immunology; Chlorocebus aethiops; Clostridium butyricum; Clostridium butyricum - immunology; Clostridium difficile; Clostridium difficile - growth & development; Clostridium difficile - immunology; Coculture Techniques; Colitis; Cytotoxicity; Data processing; Detoxification; Diarrhea; Digestive tract; Enterotoxins - genetics; Enterotoxins - immunology; Fundamental and applied biological sciences. Psychology; Hospitals; Humans; Infection; Infectious diseases; Inoculation; Medical sciences; Metabolism; Microbiology; Miscellaneous; mRNA; organic acids; Pathogens; Pharmaceuticals; probiotics; Probiotics - pharmacology; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Bacterial - chemistry; RNA, Bacterial - genetics; Secretion; Toxicity; Toxins; Vero Cells; Clostridiales; Bacteria; In vitro; Clostridiaceae; Clostridium difficile; Strain
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.033423-0

In contrast to most modern pharmaceuticals, probiotics are used in many parts of the world with little or no research data on the complex system of interactions that each strain may elicit in the human body. Research on probiotics has recently become more significant, as probiotics have begun to be prescribed by clinicians as an alternative for some gut infections, especially when antibiotics are contraindicated. This study attempted to elucidate the inhibitory interaction between the Japanese probiotic strain Clostridium butyricum MIYAIRI 588 (CBM588) and the hospital pathogen Clostridium difficile, which is responsible for a large proportion of antibiotic-associated diarrhoea and colitis. CBM588 has previously shown effectiveness against C. difficile in vivo, and here it was found that the toxicity of C. difficile in in vitro co-culture with CBM588 was greatly decreased or absent. This was dependent on the inoculation ratio and was not accounted for by the small degree of growth and mRNA inhibition observed. CBM588 and its cell-free supernatant also had no effect on toxin already secreted into the culture medium, and culture of the two strains separated by a semi-permeable membrane resulted in loss of the inhibition. Therefore, it was concluded that the detoxification probably occurred by the inhibition of toxin protein production and that this required close proximity or contact between the two species. The low-pH conditions caused by organic acid secretion were also observed to have inhibitory effects on C. difficile growth, metabolism and toxicity.