Microbiological and molecular Studies on Helicobacter species in Equine

Abstract

A total of fifty diseased horses (30 foals and 20 adults) suffering from various degrees of abdominal colicky pain and samples were taken during episodes of colic. Gastric lavage and fecal samples were collected from each horse, then lateral flow immunoassay (LF) was used as a screening test for detection of H. pylori antigen from gastric lavage and fecal samples by using monoclonal anti-bodies dependant kit. According to Moyaert et al., the PCR reaction was conducted using Helicobacter species specific primer for 16S rRNA gene, and one amplicon was chosen for sequencing, a purified PCR product was sequenced in the forward and reverse directions on an automated DNA sequencer (Applied Bio systems). A BLAST analysis was initially performed to establish sequence identity to GenBank accessions, a comparative analysis of sequences was performed using the CLUSTAL W multiple sequence alignment program and Phylogenetic analyses were done using maximum likelihood, neighbor joining and maximum parsimony in MEGA X software. Unfortunately, all (LF) tests were negative for both samples gastric lavages and faeces. Out of the 50 examined fecal samples, 12 (24%) were positive for Helicobacter species and all were detected in foal's samples, The results were obtained by PCR ,The sequence shared 100% identity to Helicobacter equorum by sequencing and phylogenetic analysis ,then