title: Necrotoxigenic Escherichia coli from sheep and goats produce a new type of cytotoxic necrotizing factor (CNF3) associated with the eae and ehxA genes.
abstract: Fecal samples from sheep and goats were screened by tissue-culture assays and PCR for the presence of necrotoxigenic Escherichia coli (NTEC) producing cytotoxic necrotizing factors (CNFs). Of the 18 NTEC strains assayed, four were positive for the cnf1 gene while 14 strains were negative for the cnf1 and cnf2 genes. All of the NTEC strains had the eae gene and most of them also carried the ehxA gene. Moreover, all the cnf1- cnf2- NTEC strains were negative for several virulence markers associated with CNF1+ or CNF2+ strains. The cnf gene present in one of these strains was sequenced and analysis of the gene product revealed a new type of CNF, which was named CNF3 (and the coding gene cnf3). Oligonucleotide primers were designed to PCR-amplify a fragment of cnf3. The results showed that all strains examined in this study, except one cnf1+strain, were cnf3+. The association of cnf3 with eae and ehxA suggests that cnf3+ NTEC strains might be pathogenic for humans.
authors.affiliation: Departament of Animal Health, Faculty of Veterinary, Complutense University of Madrid, Spain. jaorden@vet.ucm.es
authors.forename: José A
authors.initials: JA
authors.lastname: Orden
authors.forename: Gustavo
authors.initials: G
authors.lastname: Domínguez-Bernal
authors.forename: Susana
authors.initials: S
authors.lastname: Martínez-Pulgarín
authors.forename: Miguel
authors.initials: M
authors.lastname: Blanco
authors.forename: Jesús E
authors.initials: JE
authors.lastname: Blanco
authors.forename: Azucena
authors.initials: A
authors.lastname: Mora
authors.forename: Jorge
authors.initials: J
authors.lastname: Blanco
authors.forename: Jorge
authors.initials: J
authors.lastname: Blanco
authors.forename: Ricardo
authors.initials: R
authors.lastname: de la Fuente
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D003967:Diarrhea / Q000382:microbiology / Q000662:veterinary; D016521:Electrophoresis, Gel, Pulsed-Field / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015511:Goat Diseases / Q000382:microbiology*; D006041:Goats; D006460:Hemolysin Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D012756:Sheep; D012757:Sheep Diseases / Q000382:microbiology*
chemical_list: D018829:Adhesins, Bacterial; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; C060296:cytotoxic necrotizing factor type 1; C073819:eaeA protein, E coli