Abstract
Clostridium difficile infection is a toxin-mediated disease of the colon. C. difficile virulence is primarily attributed to the production of toxin A and toxin B; thus this study was aimed to investigate the effect of a range of natural products on the production and activity of C. difficile toxins in vitro. Twenty-two natural products were investigated against four C. difficile strains. The activity of products against toxins was determined using Vero and HT-29 cells cytotoxicity and neutral red uptake assays. The indirect effect of products on toxin-mediated cytotoxicity was determined using the same cell lines. The effect of seven products on toxin production by C. difficile was determined using ELISA. Zingerone (0.3 mg/ml) protected both cell lines from C. difficile cytopathic effects, confirmed by the neutral red uptake assay (P < 0.05). Three Leptospermum honeys (4% w/v), fresh onion bulb extract (12.5% v/v) and trans-cinnamaldehyde (0.005% v/v) all reduced toxin production and activity significantly (P ≤ 0.023). Garlic clove powder (4.7 mg/ml) only reduced toxin activity (P ≤ 0.047). Overall, several natural products had activity against C. difficile toxins in vitro encouraging further investigation against C. difficile toxins in vivo.
Highlights
Clostridium difficile infection (CDI) is one of the most important healthcare-associated infections worldwide and a major cause of morbidity and mortality in both the hospital and community settings[1]
We have shown recently that several natural products have antimicrobial activity against C. difficile in vitro[13]
Only minor cytopathic effect (CPE) was observed with C. difficile ATCC 43598 and HT-29 cells
Summary
Clostridium difficile infection (CDI) is one of the most important healthcare-associated infections worldwide and a major cause of morbidity and mortality in both the hospital and community settings[1]. TcdA and TcdB have different cellular receptors, in vivo potency and immunological response, and recent studies of isogenic toxin mutants in hamster and piglet models provided convincing evidence that toxin B alone is essential for CDI3,4. Secretion of these potent toxins within the gastrointestinal tract causes actin disassembly, enterocyte apoptosis, the breakdown of epithelial tight junctions and an overall loss of epithelial integrity[3]. Plant-derived compounds are considered by some consumers as a safer, less toxic and more environmentally-friendly option compared to conventional therapies They are typically multi-component in nature and the components contain different functional groups. ELISA was performed to determine the effect of these products on the production of C. difficile toxins
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