Detection of Clostridium perfringens toxinotypes, enteropathogenic E. coli, rota and corona viruses in the intestine of neonatal goat kids by molecular techniques

Abstract

The role of various pathogenic agents affecting gastro-intestinal tract of neonatal goat kids was studied. Intestinal tissue samples of 142 necropsied kids died due to enteritis, pneumo-enteritis, tapeworm’s infestations and septicemia were collected for detection of toxinotypes of Clostridium perfringens, enteropathogenic E. coli (EPEC), Group-A rotavirus (GARV) and bovine coronavirus (BCV). C. perfringens toxinotyping was done by multiplex PCR using primers for cpa, cpb, cpb2, etx and iap gene. Further, identification of EPEC was done using SYBR green based real-time PCR assay targeting bfpA gene. Detection of GARV and BCV was done by one-step RT-PCR (osRTPCR). The incidence of C. perfringens was 16.20% (47.83% toxinotype A and 52.17% toxinotype D), with b2- toxin present in 30.43% of the samples. Incidence of EPEC in 0-1 and 1–3 month diarrhoeic kids was 36.62% and 25.35%, respectively. For viral aetiology, 11.97% were positive for GARV and 9.86% positive for BCV. Mixed infection of C. perfringens and EPEC was seen in 12.68% cases, while it was 2.11% for C. perfringens + GARV and 1.40% for C. perfringens + BCV. Similarly, the mixed occurrence of EPEC+GARV was 10.56% and that of EPEC+BCV was 7.04%. The combined infection of EPEC+GARV+BCV was 0.70%. The results suggested that C. perfringens type A and type D are the common toxinotypes affecting neonatal goat kids, with β2 toxin being an additional potential virulence factor. SYBR green based real-time PCR assay can be used as quick lab-based technique for efficient screening of EPEC from enteritis affected kids.Group-A rotavirus and bovine coronavirus appeared to be instrumental in causing mixed infection enteritis in kids.