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. 1996 Jan;34(1):144–148. doi: 10.1128/jcm.34.1.144-148.1996

Genotypic and phenotypic characterization of Escherichia coli isolates from dogs manifesting attaching and effacing lesions.

M Beaudry 1, C Zhu 1, J M Fairbrother 1, J Harel 1
PMCID: PMC228748  PMID: 8748291

Abstract

Thirteen Escherichia coli isolates from dogs manifesting attaching and effacing lesions were characterized genetically with respect to the presence of the following virulence determinants associated with human enteropathogenic E. coli (EPEC): eaeA, encoding the outer membrane protein intimin; eaeB, which is necessary for inducing signal transduction; bfpA, encoding the bundle-forming pilus; and the EAF (stands for EPEC adherence factor) plasmid. These isolates were also analyzed phenotypically with respect to adherence to mammalian cells in vivo and in vitro. Nine of these 13 isolates were found to be eaeA positive by PCR: four of these nine were eaeB positive. The 5' end, but not the 3' end, of the eaeA gene was amplified by PCR when primers derived from the eaeA gene of EPEC were used. Six and eight of these 13 isolates were found to be bfpA positive and EAF positive, respectively. The bfpA gene and EAF locus were found on high-molecular-weight plasmids, whereas the eaeA and eaeB genes were chromosomally located when present. Only one canine E. coli isolate, 4221, which was positive for eaeA, eaeB, bfpA, and EAF, adhered to HEp-2 cells in a localized manner and was positive in the fluorescence actin staining test. The nine eaeA-positive isolates adhered to the mucosal surface of piglet ileal explants and induced some microvillus effacement. However, when tested in experimentally inoculated gnotobiotic piglets, isolate 4221 did not induce attaching and effacing lesions at any level of the intestinal tract. Our results indicate that canine E. coli isolates associated with attaching and effacing lesions share some properties with human EPEC but form a heterogeneous group.

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Selected References

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  1. Albert M. J., Alam K., Islam M., Montanaro J., Rahaman A. S., Haider K., Hossain M. A., Kibriya A. K., Tzipori S. Hafnia alvei, a probable cause of diarrhea in humans. Infect Immun. 1991 Apr;59(4):1507–1513. doi: 10.1128/iai.59.4.1507-1513.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albert M. J., Faruque S. M., Ansaruzzaman M., Islam M. M., Haider K., Alam K., Kabir I., Robins-Browne R. Sharing of virulence-associated properties at the phenotypic and genetic levels between enteropathogenic Escherichia coli and Hafnia alvei. J Med Microbiol. 1992 Nov;37(5):310–314. doi: 10.1099/00222615-37-5-310. [DOI] [PubMed] [Google Scholar]
  3. Beebakhee G., Louie M., De Azavedo J., Brunton J. Cloning and nucleotide sequence of the eae gene homologue from enterohemorrhagic Escherichia coli serotype O157:H7. FEMS Microbiol Lett. 1992 Feb 1;70(1):63–68. doi: 10.1016/0378-1097(92)90563-4. [DOI] [PubMed] [Google Scholar]
  4. Broes A., Drolet R., Jacques M., Fairbrother J. M., Johnson W. M. Natural infection with an attaching and effacing Escherichia coli in a diarrheic puppy. Can J Vet Res. 1988 Apr;52(2):280–282. [PMC free article] [PubMed] [Google Scholar]
  5. Broes A., Fairbrother J. M., Mainil J., Harel J., Lariviere S. Phenotypic and genotypic characterization of enterotoxigenic Escherichia coli serotype O8:KX105 and O8:K"2829" strains isolated from piglets with diarrhea. J Clin Microbiol. 1988 Nov;26(11):2402–2409. doi: 10.1128/jcm.26.11.2402-2409.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cantey J. R., Moseley S. L. HeLa cell adherence, actin aggregation, and invasion by nonenteropathogenic Escherichia coli possessing the eae gene. Infect Immun. 1991 Nov;59(11):3924–3929. doi: 10.1128/iai.59.11.3924-3929.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coté S., Harel J., Higgins R., Jacques M. Resistance to antimicrobial agents and prevalence of R plasmids in Pasteurella multocida from swine. Am J Vet Res. 1991 Oct;52(10):1653–1657. [PubMed] [Google Scholar]
  8. Daigle F., Harel J., Fairbrother J. M., Lebel P. Expression and detection of pap-, sfa-, and afa-encoded fimbrial adhesin systems among uropathogenic Escherichia coli. Can J Microbiol. 1994 Apr;40(4):286–291. doi: 10.1139/m94-046. [DOI] [PubMed] [Google Scholar]
  9. Donnenberg M. S., Girón J. A., Nataro J. P., Kaper J. B. A plasmid-encoded type IV fimbrial gene of enteropathogenic Escherichia coli associated with localized adherence. Mol Microbiol. 1992 Nov;6(22):3427–3437. doi: 10.1111/j.1365-2958.1992.tb02210.x. [DOI] [PubMed] [Google Scholar]
  10. Donnenberg M. S., Kaper J. B. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immun. 1991 Dec;59(12):4310–4317. doi: 10.1128/iai.59.12.4310-4317.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Donnenberg M. S., Kaper J. B. Enteropathogenic Escherichia coli. Infect Immun. 1992 Oct;60(10):3953–3961. doi: 10.1128/iai.60.10.3953-3961.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Donnenberg M. S., Yu J., Kaper J. B. A second chromosomal gene necessary for intimate attachment of enteropathogenic Escherichia coli to epithelial cells. J Bacteriol. 1993 Aug;175(15):4670–4680. doi: 10.1128/jb.175.15.4670-4680.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Drolet R., Fairbrother J. M., Harel J., Hélie P. Attaching and effacing and enterotoxigenic Escherichia coli associated with enteric colibacillosis in the dog. Can J Vet Res. 1994 Apr;58(2):87–92. [PMC free article] [PubMed] [Google Scholar]
  14. Foubister V., Rosenshine I., Donnenberg M. S., Finlay B. B. The eaeB gene of enteropathogenic Escherichia coli is necessary for signal transduction in epithelial cells. Infect Immun. 1994 Jul;62(7):3038–3040. doi: 10.1128/iai.62.7.3038-3040.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Frankel G., Candy D. C., Everest P., Dougan G. Characterization of the C-terminal domains of intimin-like proteins of enteropathogenic and enterohemorrhagic Escherichia coli, Citrobacter freundii, and Hafnia alvei. Infect Immun. 1994 May;62(5):1835–1842. doi: 10.1128/iai.62.5.1835-1842.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Girón J. A., Ho A. S., Schoolnik G. K. An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science. 1991 Nov 1;254(5032):710–713. doi: 10.1126/science.1683004. [DOI] [PubMed] [Google Scholar]
  17. Gross R. J., Rowe B. Escherichia coli diarrhoea. J Hyg (Lond) 1985 Dec;95(3):531–550. doi: 10.1017/s0022172400060666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Harel J., Lapointe H., Fallara A., Lortie L. A., Bigras-Poulin M., Larivière S., Fairbrother J. M. Detection of genes for fimbrial antigens and enterotoxins associated with Escherichia coli serogroups isolated from pigs with diarrhea. J Clin Microbiol. 1991 Apr;29(4):745–752. doi: 10.1128/jcm.29.4.745-752.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Janke B. H., Francis D. H., Collins J. E., Libal M. C., Zeman D. H., Johnson D. D. Attaching and effacing Escherichia coli infections in calves, pigs, lambs, and dogs. J Vet Diagn Invest. 1989 Jan;1(1):6–11. doi: 10.1177/104063878900100104. [DOI] [PubMed] [Google Scholar]
  20. Jerse A. E., Gicquelais K. G., Kaper J. B. Plasmid and chromosomal elements involved in the pathogenesis of attaching and effacing Escherichia coli. Infect Immun. 1991 Nov;59(11):3869–3875. doi: 10.1128/iai.59.11.3869-3875.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jerse A. E., Kaper J. B. The eae gene of enteropathogenic Escherichia coli encodes a 94-kilodalton membrane protein, the expression of which is influenced by the EAF plasmid. Infect Immun. 1991 Dec;59(12):4302–4309. doi: 10.1128/iai.59.12.4302-4309.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jerse A. E., Yu J., Tall B. D., Kaper J. B. A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7839–7843. doi: 10.1073/pnas.87.20.7839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Knutton S., Baldwin T., Williams P. H., McNeish A. S. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun. 1989 Apr;57(4):1290–1298. doi: 10.1128/iai.57.4.1290-1298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Law D. Adhesion and its role in the virulence of enteropathogenic Escherichia coli. Clin Microbiol Rev. 1994 Apr;7(2):152–173. doi: 10.1128/cmr.7.2.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Le Bouguenec C., Archambaud M., Labigne A. Rapid and specific detection of the pap, afa, and sfa adhesin-encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction. J Clin Microbiol. 1992 May;30(5):1189–1193. doi: 10.1128/jcm.30.5.1189-1193.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Levine M. M. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis. 1987 Mar;155(3):377–389. doi: 10.1093/infdis/155.3.377. [DOI] [PubMed] [Google Scholar]
  27. Levine M. M., Prado V., Robins-Browne R., Lior H., Kaper J. B., Moseley S. L., Gicquelais K., Nataro J. P., Vial P., Tall B. Use of DNA probes and HEp-2 cell adherence assay to detect diarrheagenic Escherichia coli. J Infect Dis. 1988 Jul;158(1):224–228. doi: 10.1093/infdis/158.1.224. [DOI] [PubMed] [Google Scholar]
  28. Moxley R. A., Francis D. H. Natural and experimental infection with an attaching and effacing strain of Escherichia coli in calves. Infect Immun. 1986 Aug;53(2):339–346. doi: 10.1128/iai.53.2.339-346.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nataro J. P., Baldini M. M., Kaper J. B., Black R. E., Bravo N., Levine M. M. Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. J Infect Dis. 1985 Sep;152(3):560–565. doi: 10.1093/infdis/152.3.560. [DOI] [PubMed] [Google Scholar]
  30. Pohl P. H., Peeters J. E., Jacquemin E. R., Lintermans P. F., Mainil J. G. Identification of eae sequences in enteropathogenic Escherichia coli strains from rabbits. Infect Immun. 1993 May;61(5):2203–2206. doi: 10.1128/iai.61.5.2203-2206.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Robins-Browne R. M., Tokhi A. M., Adams L. M., Bennett-Wood V., Moisidis A. V., Krejany E. O., O'Gorman L. E. Adherence characteristics of attaching and effacing strains of Escherichia coli from rabbits. Infect Immun. 1994 May;62(5):1584–1592. doi: 10.1128/iai.62.5.1584-1592.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rosenshine I., Finlay B. B. Exploitation of host signal transduction pathways and cytoskeletal functions by invasive bacteria. Bioessays. 1993 Jan;15(1):17–24. doi: 10.1002/bies.950150104. [DOI] [PubMed] [Google Scholar]
  33. Scaletsky I. C., Silva M. L., Trabulsi L. R. Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells. Infect Immun. 1984 Aug;45(2):534–536. doi: 10.1128/iai.45.2.534-536.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schauer D. B., Falkow S. Attaching and effacing locus of a Citrobacter freundii biotype that causes transmissible murine colonic hyperplasia. Infect Immun. 1993 Jun;61(6):2486–2492. doi: 10.1128/iai.61.6.2486-2492.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tzipori S., Gibson R., Montanaro J. Nature and distribution of mucosal lesions associated with enteropathogenic and enterohemorrhagic Escherichia coli in piglets and the role of plasmid-mediated factors. Infect Immun. 1989 Apr;57(4):1142–1150. doi: 10.1128/iai.57.4.1142-1150.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tzipori S., Robins-Browne R. M., Gonis G., Hayes J., Withers M., McCartney E. Enteropathogenic Escherichia coli enteritis: evaluation of the gnotobiotic piglet as a model of human infection. Gut. 1985 Jun;26(6):570–578. doi: 10.1136/gut.26.6.570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ulshen M. H., Rollo J. L. Pathogenesis of escherichia coli gastroenteritis in man--another mechanism. N Engl J Med. 1980 Jan 10;302(2):99–101. doi: 10.1056/NEJM198001103020207. [DOI] [PubMed] [Google Scholar]
  38. Wolf M. K., Andrews G. P., Fritz D. L., Sjogren R. W., Jr, Boedeker E. C. Characterization of the plasmid from Escherichia coli RDEC-1 that mediates expression of adhesin AF/R1 and evidence that AF/R1 pili promote but are not essential for enteropathogenic disease. Infect Immun. 1988 Aug;56(8):1846–1857. doi: 10.1128/iai.56.8.1846-1857.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wolf M. K., Boedeker E. C. Cloning of the genes for AF/R1 pili from rabbit enteroadherent Escherichia coli RDEC-1 and DNA sequence of the major structural subunit. Infect Immun. 1990 Apr;58(4):1124–1128. doi: 10.1128/iai.58.4.1124-1128.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yu J., Kaper J. B. Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157:H7. Mol Microbiol. 1992 Feb;6(3):411–417. doi: 10.1111/j.1365-2958.1992.tb01484.x. [DOI] [PubMed] [Google Scholar]
  41. Zhu C., Harel J., Jacques M., Desautels C., Donnenberg M. S., Beaudry M., Fairbrother J. M. Virulence properties and attaching-effacing activity of Escherichia coli O45 from swine postweaning diarrhea. Infect Immun. 1994 Oct;62(10):4153–4159. doi: 10.1128/iai.62.10.4153-4159.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zhu C., Harel J., Jacques M., Fairbrother J. M. Interaction with pig ileal explants of Escherichia coli O45 isolates from swine with postweaning diarrhea. Can J Vet Res. 1995 Apr;59(2):118–123. [PMC free article] [PubMed] [Google Scholar]

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