Evaluation of toxinogenesis of Clostridium perfringens type D isolates in three kinds of culture media

Clostridium perfringens (C. perfringens) causes many diseases in humans and livestock. The kind of secreted toxins is the basis of C. perfringens classification. One of the most important toxins in the epsilon toxin produced by C. perfringens type D. The purpose of this study is evaluating the produce of C. perfringens type D epsilon toxin in three types of culture medium (A, B, and C). The differences between these three media are the type of peptone and commercial liver powder as the main nutrient and protein supplier. In this study, C. perfringens type D isolates cultured in three different nutrient culture mediums, after growth assurance, the suspensions used for evaluation of toxicity by Minimum Lethal Dose (MLD) test, total protein content and ELISA assays. The presence of alpha and epsilon toxins in media were confirmed by ELISA assay. The mean MLD values were 121, 89, and 98 MLD/ml in A, B, and C media, respectively. The mean total protein contents were 53, 86.7, and 104.4 mg/ml in A, B, and C, respectively. Statistical analysis showed there is a direct correlation between the results of the total protein contents and MLD values. The results showed the three types of culture media used in the growth and power of C. perfringens type D to produce epsilon toxin was similar, in other words, they can be used as alternatives.