Toxicity effects of alpha (α) and epsilon (ε) toxins produced by Clostridium perfringens type D

Subject Areas : Bacteriology

Mahnaz Karimi ¹ (M.Sc., Department of Microbiology, Jahrom Branch, Islamic Azad University Jahrom, Iran.)
Mehrdad Shamsaddini Bafti ² (Assistant Professor, Department of Research and Development of Biological Products, Razi Vaccine and Serum Research Institute, Kerman, Iran.)
Majid Ezatkhah ³ (Lecturer, Razi Vaccine & Serum Research Institute, Kerman, Iran.)
Mohammad Kargar ⁴ (Associate Professor, Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.)

Received: 2013-09-25 Accepted : 2014-01-17 Published : 2014-04-01

Keywords: Gene expression, toxin, Clostridium perfringens, Culture media, Liver powder,

Abstract :

Background and Objectives: Clostridium perfringens is one of the most pathogenic species in the clostridium genus. Alpha (α) and epsilon (ε) are the main toxins of this bacteria. This study aimed to express the genes encoding alpha and epsilon toxins of C. perfringens type D in two media (in the base medium and the base medium containing liver powder) and to analyze their toxicity. Materials and Methods: The standard strain was inoculated in the base enrichment medium, which half of them were enriched by powder liver. Then, RNAs were extracted and after converting to cDNA using double polymerase chain reaction, the gene expression was investigated on gels. The level of protein expression was measured by spectrophotometery and the cell toxicity of these proteins was determined by minimum lethal dose (MLD) assay. Results: The electrophoresis of PCR products showed two bands, 324 and 625, which represented successful expression of the genes encoding of alpha and epsilon toxins in both media. The mean cell cytotoxicity of the proteins expressed in the base medium and the base medium containing liver powder were measured 1/4000 and 1/4333, respectively. The mean protein production in the base medium and the base medium containing liver powder were measured 94.39 and 48.01 mg/ml. Conclusions: This study showed that these genes are expressed in the presence of liver powder. Furthermore, based on the MLD assay in these media, although these genes expressed the genes in the presence of liver powder, this additives did not had any effects on their toxicity.