Clostridium perfringens is a gram-positive, spore-forming bacillus, and is a causative agent of foodborne infection, antibiotic-associated diarrhoea and sporadic diarrhoea in humans. In cases of antibiotic-associated and sporadic diarrhoea, C. perfringens colonises the intestine, proliferates and causes disease. However, bacterial colonisation of the intestine is not considered necessary in the pathogenesis of foodborne illness, because such pathogenesis can be explained by anchorage-independent production of diarrhoeic toxin by the bacterium in the intestine. In this study, we used an in vitro adherence assay to examine the adherence of C. perfringens spores to human intestinal Caco-2 cells. Adherence of spores from isolates of foodborne illness and nosocomial infection was observed within 15 min, and plateaued 60 min after inoculation. Electron microscopy revealed a tight association of spores with the surface of Caco-2 cells. The adherence of vegetative cells could not be confirmed by the same method, however. These results suggest that C. perfringens spores may adhere to intestinal epithelial cells in vivo, although its biological significance remains to be determined.

Sublethal heating of spores has long been known to stimulate or activate germination; however, the underlying mechanisms are not yet fully understood. In this study, the entire germination-to-outgrowth process of spores from Clostridium perfringens, an anaerobic sporeformer, was visualized at single-cell resolution. Quantitative analysis revealed that sublethal heating significantly reduces the time from completion of germination to the beginning of the first cell division, indicating that sublethal heating of C. perfringens spores not only sensitizes the responsiveness of germinant receptors but also directly or indirectly facilitates multiple steps during the bacterial regrowth process.

Spores are a highly resistant form of bacteria and are capable of surviving various environmental stresses. When food is contaminated with spores, it can result in food spoilage and food-borne illness. Although it is important to develop procedures to control the germination and outgrowth of food-contaminating spores, there is limited biological information about the spores of anaerobic Clostridium spp. In this mini-review, recent advances in the biological behaviors of Clostridium spp. spores are summarized, and the possible burden in food hygiene induced by putative damaged-but-viable spores is discussed. Concerted efforts to elucidate the biological specificity of spores are needed to develop new control measures applicable to a variety of hygienic processes.