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Background   Clostridium perfringens  type A food poisoning is caused by enterotoxigenic  C. perfringens  type A isolates that typically possess high spore heat-resistance. The molecular basis for  C. perfringens  spore heat-resistance remains unknown. In the current study, we investigated the role of small, acid-soluble spore proteins (SASPs) in heat-resistance of spores produced by  C. perfringens  food poisoning isolates.    Results  Our current study demonstrated the presence of all three SASP-encoding genes ( ssp1 ,  2  and  3 ) in five surveyed  C. perfringens  clinical food poisoning isolates. β-Glucuronidase assay showed that these  ssp  genes are expressed specifically during sporulation. Consistent with these expression results, our study also demonstrated the production of SASPs by  C. perfringens  food poisoning isolates. When the heat sensitivities of spores produced by a  ssp3  knock-out mutant of a  C. perfringens  food poisoning isolate was compared with that of spores of the wild-type strain, spores of the  ssp3  mutant were found to exhibit a lower decimal reduction value ( D  value) at 100°C than exhibited by the spores of wild-type strain. This effect was restored by complementing the  ssp3  mutant with a recombinant plasmid carrying wild-type  ssp3 , suggesting that the observed differences in  D  values between spores of wild-type versus  ssp3  mutant was due to the specific inactivation of  ssp3 . Furthermore, our DNA protection assay demonstrated that  C. perfringens  SASPs can protect DNA from DNase I digestion.    Conclusion  The results from our current study provide evidences that SASPs produced by  C. perfringens  food poisoning isolates play a role in protecting their spores from heat-damage, which is highly significant and relevant from a food safety perspective. Further detailed studies on mechanism of action of SASPs from  C. perfringens  should help in understanding the mechanism of protection of  C. perfringens  spores from heat-damage.

Investigating the role of small, acid-soluble spore proteins (SASPs) in the resistance of Clostridium perfringens spores to heat