Molecular characterization of clostridium perfringens toxin types isolated from sheep of Andhra Pradesh

Abstract

Multiplex PCR was standardized using specific primers for alpha, beta and epsilon toxin genes of reference strains of C. perfringens. The average detection limit of the test was found to be 1- 1.5 x 104 cfu ml -1 with purified DNA extracted from the reference strains. A total of 208 clinical samples which includes 140 from ET suspected lambs and 68 from healthy slaughtered animals were subjected to multiplex PCR screening. The bacterial lysate of 24h broth culture of clinical samples were found suitable for effective screening for C. perfringens types in clinical samples. newlineOut of 208 samples tested by multiplex PCR from ET suspected and healthy lambs, 181 were positive for C. perfringens with overall prevalence of 87.01 per cent. Out of 140 samples from ET suspected animals, 129 (92.14%) were positive, whereas 68 samples from healthy animals revealed 52 (76.47%) positives. The toxin genotyping by multiplex PCR screening showed Clostridium perfringens type A as the predominant type with prevalence of 69.61 per cent. The prevalence of 92.30 per cent of C. perfringens type A healthy animals was found to be highly significant compared with ET suspected group .C. perfringens type C is found to be the least prevalent (11.04%) among all the C. perfringens types recovered in the present study. A prevalence of C. perfringens type D was found to be 19.33 per cent with 24.82 per cent prevalence of in ET suspected animals which is significantly higher than healthy animals (5.76%). newline newline