Graduate Thesis Or Dissertation
 

Characterization of germination of Clostridium perfringens spores from various sources

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  • Clostridium perfringens is a pathogenic anaerobic bacterium able to produce more than 17 toxins, allowing C. perfringins to cause a wide variety of diseases in humans and animals. Beside toxins production, C. perfringens able to form a highly resistance spores can survive in the environments for years. These spores are the infectious cell morphotype, and in presence of favorable condition, these spores germinate and return to active growth to cause disease. Spore germination is an early and essential stage in the progression of C. perfringens infection in human and animal. It can be initiated by a variety of chemicals, including nutrients, cationic surfactants, and enzymes termed germinant. Germination of Clostridum species has been less well studied than Bacillus species. However, recent findings have identified the germinants of spores of C. perfringens food poisoning (FP) and non-food borne (NFB) isolates. The first focus of this project was to compare the nutrient and non-nutrient induced germination of spores of FP versus NFB isolates. The result showed that spores of FP isolates can germinate with KCl, L-asparagine and a mixture of KCl and L-asparagine (AK), while spores of NFB isolates germinate well only with the AK mixture. While dodecylamine is a universal germinant for spores of both FP and NFB isolates, a 1:1 chelate of Ca²⁺ and dipicolinic acid (Ca-DPA) can induce germination of spores of most FP isolates but none of NFB isolates. These results suggest a possible difference between spore germination mechanism in FP versus NFB isolates and this might be, at least in part, due to the differences in germination machinery. The second focus of this study was to investigate the germination requirements of spores of C. perfringens animal isolates (AI). Result from this study found that although AI spores germinated poorly in nutrient broth, they germinated well in tissue culture media, specially RPMI 1640. Most notably, was the ability of L-threonine to trigger germination of most AI spores, followed by L-lysine and to a lesser extent L-alanine. There was no correlation in the germinant requirements and the source of isolation of AI. Interestingly, this study also reports novel germinants for FP and NFB spores which include L-glutamine and L-aspartic acid and to a lesser extent L-lysine and L-arginine. Collectively, the present study contributes to the understanding of the germinant specificity of spores of C. perfringens AI and the differential germinant requirements between FP and NFB spores.
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