Production of epsilon toxin secreted by Clostridium perfringens type D under different culture conditions

Clostridium perfringens is a Gram-positive, rod-shaped, anaerobic, spore-forming pathogenic bacterium that causes various diseases such as dysentery and gas gangrene in animals and humans. Epsilon () toxin secreted by C. perfringens type B and type D causes a worldwide disease known as enterotoxemia, which occurs when the toxin is absorbed from the intestines in sheep, goats and cattle. Owing to the rapid and severe course of the disease, vaccination is important to fight the disease. One way to control enterotoxemic diseases is vaccination using inactivated toxins called "toxoids" or inactivated bacterial cells. Immunization with C. perfringens type D toxoid preparations has made it possible to control the enterotoxemic diseases caused by this strain to a certain extent. To increase the potency of this toxoid preparation, it is necessary to obtain the toxin that provides a high degree of antibody response. In previous studies, although -toxin was obtained, the toxin levels were low. In this study, the effect of temperature, pH, zinc (ZnSO4) on epsilon toxin production by C. perfringens type D were investigated. The growth dynamics of bacteria under such varying culture conditions were studied to understand whether any relationship exists between the density of bacteria in the culture medium and epsilon toxin production. Quantitative and qualitative determination of the toxin was determined by SDS-PAGE analysis. Our findings showed optimum production of -toxin was observed at 37 oC, pH 6.8 and there was no correlation between bacterial biomass and toxin production.