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Clostridium botulinum  is a dangerous pathogen that forms the highly potent botulinum toxin, which when ingested causes a deadly neuroparalytic disease. The closely related  Clostridium sporogenes  is occasionally pathogenic, frequently associated with food spoilage and regarded as the non-toxigenic equivalent of Group I  C. botulinum . Both species form highly resistant spores that are ubiquitous in the environment and which, under favourable growth conditions germinate to produce vegetative cells. To improve the control of botulinum neurotoxin-forming clostridia, it is imperative to comprehend the mechanisms by which spores germinate. Germination is initiated following the recognition of small molecules (germinants) by a specific germinant receptor (GR) located in the spore inner membrane. The present study precisely defines clostridial GRs, germinants and co-germinants. Group I  C. botulinum  ATCC3502 contains two tricistronic and one pentacistronic GR operons, while  C. sporogenes  ATCC15579 has three tricistronic and one tetracistronic GR operons. Insertional knockout mutants, allied with characterisation of recombinant GRs shows for the first time that amino acid stimulated germination in  C. botulinum  requires two tri-cistronic encoded GRs which act in synergy and cannot function individually. Spore germination in  C. sporogenes  requires one tri-cistronic GR. Two other GRs form part of a complex involved in controlling the rate of amino-acid stimulated germination. The suitability of using  C. sporogenes  as a substitute for  C. botulinum  in germination studies and food challenge tests is discussed.

plos pathogens

Functional Characterisation of Germinant Receptors in Clostridium botulinum and Clostridium sporogenes Presents Novel Insights into Spore Germination Systems