title: A PCR technique to detect enterotoxigenic and verotoxigenic Escherichia coli in boar semen samples.
abstract: In semen, bacteria's isolation from a pure culture is complex, laborious and easily alterable by the presence of antibiotics and inhibitors. We developed a PCR technique to detect the presence of the enterotoxigenic (ETEC) and verotoxigenic Escherichia coli (VTEC) (strains with high prevalence in the swine industry) in semen by adapting the protocols developed by Zhang et al. (2007) and Yilmaz et al. (2006). We artificially inoculated extended semen samples at different infective concentrations of bacteria (from 10(2) to 10(8) bacteria ml(-1)) with two enterotoxigenic and verotoxigenic strains, and performed two multiplex and one conventional PCR. This technique proved to be a quick, useful and reliable tool to detect the presence of ETEC and VTEC up to an infective dose of 10(5) bacteria ml(-1) in semen.
authors.affiliation: Biotecnologia de la Reproducció Animal i Humana (TechnoSperm), Departament de Biologia, Facultat de Ciències, Universitat de Girona, E-17071 Girona, Catalonia, Spain. eva.bussalleu@udg.edu
authors.forename: E
authors.initials: E
authors.lastname: Bussalleu
authors.forename: E
authors.initials: E
authors.lastname: Pinart
authors.forename: M
authors.initials: M
authors.lastname: Yeste
authors.forename: M
authors.initials: M
authors.lastname: Briz
authors.forename: S
authors.initials: S
authors.lastname: Sancho
authors.forename: E
authors.initials: E
authors.lastname: Torner
authors.forename: S
authors.initials: S
authors.lastname: Bonet
mesh_terms: D000818:Animals; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics*; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D005798:Genes, Bacterial / Q000235:genetics; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012661:Semen / Q000382:microbiology*; D012680:Sensitivity and Specificity; D054323:Shiga-Toxigenic Escherichia coli / Q000235:genetics*; D013552:Swine / Q000382:microbiology; D013553:Swine Diseases / Q000382:microbiology*