Study on in-vitro biochemical growth characterization and assessment of hemolytic toxin of Clostridium perfringens type B and D.

The present study was conducted to identify the biochemical characterization, evaluation of growth and hemolytic properties of Clostridium perfringens (C. perfringens) type B and D in different growth media and physical environment. The reinforced clostridial (RC) and thioglycolate (TG) growth media were observed better for the growth and hemolysin production 12 h post-incubation at 37 degrees C (P<0.05). Time, temperature and buffer also illustrated a significant (P<0.05) relationship on the hemolytic activity of proto-toxin of C. perfringens types B and D. The trypsinization of proto-toxin greatly enhanced hemolytic activity and 1% trypsin in physiological saline at 37 degrees C for one hour showed maximum hemolytic units (P<0.05). The reinforced clostridial and thioglycolate growth media were found better for the cultivation of C. perfringens and enhanced hemolytic activity with 1% trypsinization. Present results could be helpful in the preparation of an effective bivalent vaccine against enterotoxaemia and lamb dysentery disease of sheep and goat.