Molecular pathogenesis of enterotoxaemia in neonatal goat kids

Abstract

In this study, a total population of 1156 neonatal goat kids was examined for clinical diarrhoea newlineand 134 necropsied were examined for enteritis, pneumo-enteritis, tapeworms/round worm infestations newlineand septicemia, suspectedly died due to enterotoxemia from January, 2015 to December, 2016. A total newlineof 238 diarrhoeic fecal and 84 intestinal loop samples were collected from ICAR-CIRG, Makhdoom, newlineMathura (UP) and spontaneous outbreaks of Uttar Pradesh and Rajasthan States of India. Isolation, newlineidentification and molecular detection of toxinotypes of Clostridium perfringens were performed in newline238 diarrhoeic samples and 84 intestinal loop samples for confirmatory diagnosis of enterotoxemia. newlineDiarrhoeic samples and intestinal contents were used for isolation in Robertson cooked meat media newline(RCMM) and subsequently inoculated in 5% defibrinated sheep blood brucella agar with vitamin K1, newlinehemin and selective clostridial supplements (CLS-BBA) and egg yolk agar (EYA) under anaerobic newlineconditions. Toxinotyping were done by multiplex PCR using toxin genes viz., cpa, cpb, cpb2, etx and newlineiap. After confirmatory diagnosis, molecular characterization for different isolates was done by PCR newlinecloning and gene sequencing. The etx-full gene cloned in TA vector for various field isolates was newlinesequenced by using Bigdye® newline terminator v1.1 cycle sequencing kit (Applied biosystems). Phylogenetic newlineanalysis was conducted by minimum evolution tree analysis. Experimental study using characterized newlineisolate (CIRG-1816) was undertaken to study molecular and gene expression profiles. The inoculum newlinewas prepared in RCMM and culture supernatant (CS) containing epsilon toxin was trypsinized and newlinetitrated in mouse. Twenty male Jakhrana goat kids of 0-3 month s age weighing 9-12 kg were divided newlinein four equal groups (n=5). Group I was treated with CS, group II with washed cells (WAC), group III newlinewith whole culture (WC) and group IV with only RCM medium (control). By laparotomy, 20% starch newlinesolution was infused into abomasum and 300 ml of inoculum into duo