Molecular detection of clostridium species in beef obtained from retail shops in North West Province, South Africa

Abstract

Clostridium species cause diseases in humans that result from consumption of undercooked beef. The objectives of this study were to isolate Clostridium species from beef then the detection of the presence of tpi housekeeping gene as well as determination of the antibiotic resistant profile of the isolates. Twenty six (26) beef samples were bought from butcheries, supermarkets and street vendors. The samples were analyzed for the characteristics of Clostridium species and a total of 78 presumptive isolates were subjected to Gram-staining, catalase test, API 20A sugar fermentation profiles 16S rRNA and tpi species specific PCR analysis. Susceptibility profiles to 8 antibiotics were determined and antibiotic resistance patterns were compiled. Large proportions (93.3%-100%) of the isolates were penicillin, vancomycin and erythromycin resistant. PCR were performed to amplify species-specific 16S rRNA gene to confirm the identity of the isolates and 44.7% of the isolates were positively identified as Clostridium species. PCR were performed to amplify tpi housekeeping gene fragments. The tpi housekeeping gene produced amplicons of 501bp after PCR amplification and 19% of the isolates possess tpi housekeeping gene which confirmed the presence of Clostridium species in beef.