Molecular Characterization and Virulent Gene Detection of Clostridium perfringens from Necrotic Enteritis Cases in Kadaknath Fowl

Abstract

Enteric diseases are one of the most important problems in the poultry industry leading to production losses, high mortality and risk of contamination of poultry products for human consumption. Necrotic enteritis (NE) is one of the most clinically associated bacterial enteric disease of poultry caused by Clostridium perfringens, which is an anaerobic, gram-positive bacteria. In present study, 08 Kadaknath birds, which showed signs of inappetence and diarrhea, were presented for necropsy. Gross examination of the fowls revealed necrotic to ulcerative lesions in intestine. Clostridium perfringens organisms were isolated from intestine after anaerobic incubation. Initial enrichment of intestinal samples were done in Robertson’s cooked meat media (RCMM) and streaked on perfringens agar base with T.S.C supplements and for further confirmation, these colonies were streaked on egg yolk agar for lecithinase activity. The isolates were confirmed by 16S rRNA gene specific PCR and further processed for necrotic enteritis B like toxin (NetB) gene. Out of 08 intestinal samples processed, 04 were found positive for C. perfringens by phenotypic and genotypic confirmation. All confirmed C. perfringens isolates were processed for NetB gene and out of 4 samples, 01 was positive for NetB gene. This study further pointed out that C. perfringens NetB toxin is a one of the new key virulent factor for the development of NE in poultry birds.