1887

Abstract

Attaching and effacing (AEEC) strains isolated from diarrhoeic lambs and goat kids were characterized for intimin () and EspB () gene subtypes by PCR and sequencing, and for genetic relatedness by PFGE. Fifty (23 ovine and 27 caprine) AEEC strains of 398 (246 ovine and 152 caprine) analysed were detected by colony blot hybridization. These strains were epidemiologically unrelated since they were isolated from different outbreaks of neonatal diarrhoea over a long period. Ovine AEEC strains belonged to serogroups O2, O4, O26, O80, O91 or were untypable, and caprine strains belonged to serogroups O3, O153 and O163. Two intimin subtypes were detected among the ovine and caprine strains studied. Most of the strains (43/50) had the β type intimin gene, but seven ovine strains possessed a variant γ type intimin gene (γ). Analysis of deduced amino acid sequences of the gene revealed that the sequences of β intimin of ovine and caprine strains were virtually identical to those of β intimin of rabbit EPEC, human EPEC clone 2 and swine AEEC, whereas the γ intimin present in seven ovine strains had 75–76% identity with γ intimin of human EHEC clone 1 strains, and 96% of identity with intimin of the human EHEC strain 95NR1 of serotype O111:H–. A PCR test was developed to identify the three different gene subtypes, of human EPEC clone 1 (α), of human EHEC clone 1 (γ) and of rabbit EPEC and human EPEC clone 2 (β). There was close correlation between the intimin β type and the β gene subtype in the ovine and caprine AEEC strains. The seven ovine strains possessing the γ intimin gene possessed the α gene subtype. None of the strains studied possessed the γ gene found in human O157:H7 EHEC strains. PFGE analysis of genomic DNA of selected strains showed a great diversity among strains. Cluster analysis of PFGE patterns showed greater divergence between strains with the γ intimin gene than between strains with the β intimin gene. This study showed that most of the AEEC strains isolated from diarrhoeic lambs and goat kids possessed β intimin and genes identical to those of rabbit EPEC, and they may be associated with enteric disease in small ruminants.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-8-2341
2001-08-01
2020-01-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/8/1472341a.html?itemId=/content/journal/micro/10.1099/00221287-147-8-2341&mimeType=html&fmt=ahah

References

  1. Abe A., Kenny B., Stein M., Finlay B. B.. 1997; Characterization of two virulence proteins secreted by rabbit enteropathogenic Escherichia coli , EspA and EspB, whose maximal expression is sensitive to host body temperature. Infect Immun65:3547–3555
    [Google Scholar]
  2. Adu-Bobie J., Frankel G., Bain C., Goncalves A. G., Trabulsi L. R., Douce G., Knutton S., Dougan G.. 1998; Detection of intimins alpha, beta, gamma, and delta, four intimin derivatives expressed by attaching and effacing microbial pathogens. J Clin Microbiol36:662–668
    [Google Scholar]
  3. Agin T. S., Wolf M. K.. 1997; Identification of a family of intimins common to Escherichia coli causing attaching-effacing lesions in rabbits, humans, and swine. Infect Immun65:320–326
    [Google Scholar]
  4. Agin T. S., Cantey J. R., Boedeker E. C., Wolf M. K.. 1996; Characterization of the eae A gene from rabbit enteropathogenic Escherichia coli strain RDEC-1 and comparison to other eaeA genes from bacteria that cause attaching-effacing lesions. FEMS Microbiol Lett144:249–258[CrossRef]
    [Google Scholar]
  5. Albert M. J., Faruque S. M., Ansaruzzaman M., Islam M. M., Haider K., Alam L., Kabir I., Robins-Browne R.. 1992; Sharing of virulence-associated properties at the phenotypic and genetic levels between enteropathogenic Escherichia coli and Hafnia alvei . J Med Microbiol37:310–314[CrossRef]
    [Google Scholar]
  6. An H., Fairbrother J. M., Dubreuil J. D., Harel J.. 1997; Cloning and characterization of the eae gene from a dog attaching and effacing Escherichia coli strain 4221. FEMS Microbiol Lett148:239–245[CrossRef]
    [Google Scholar]
  7. Arbeit R. D., Arthur M., Dunn R., Kim C., Selander R. K., Goldstein R.. 1990; Resolution of recent evolutionary divergence among Escherichia coli from related lineages: the application of pulsed field electrophoresis to molecular epidemiology. J Infect Dis161:230–235[CrossRef]
    [Google Scholar]
  8. Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., Struhl K.. 1991; Current Protocols in Molecular Biology New York: Wiley;
    [Google Scholar]
  9. Beaudry M., Zhu C., Fairbrother J. M., Harel J.. 1996; Genotypic and phenotypic characterization of Escherichia coli isolates from dogs manifesting attaching and effacing lesions. J Clin Microbiol34:144–148
    [Google Scholar]
  10. Beutin L., Geiger D., Zimmermann S., Karch H.. 1996; Virulence markers of shiga-like toxin-producing Escherichia coli strains originating from healthy domestic animals of different species. J Clin Microbiol33:631–635
    [Google Scholar]
  11. Blanco J., Cid D., Blanco J. E., Blanco M., Ruiz Santa Quiteria J. A., De la Fuente R.. 1996; Serogroups, toxins and antibiotic resistance of Escherichia coli strains isolated from diarrhoeic lambs in Spain. Vet Microbiol49:209–217[CrossRef]
    [Google Scholar]
  12. Broes A., Drolet R., Jacques M., Fairbrother J. M., Johnson W. M.. 1988; Natural infection with an attaching and effacing Escherichia coli in a diarrheic puppy. Can J Vet Res52:280–282
    [Google Scholar]
  13. Cantey J. R., Blake R. K.. 1977; Diarrhoea due to Escherichia coli in the rabbit: a novel mechanism. J Infect Dis135:454–462[CrossRef]
    [Google Scholar]
  14. China B., Pirson V., Mainil J.. 1998; Prevalence and molecular typing of attaching and effacing Escherichia coli among calf populations in Belgium. Vet Microbiol63:249–259[CrossRef]
    [Google Scholar]
  15. China B., Goffaux F., Pirson V., Mainil J.. 1999a; Comparison of eae , tir , espA , and espB genes of bovine and human attaching and effacing Escherichia coli by multiplex polymerase chain reaction. FEMS Microbiol Lett178:177–182[CrossRef]
    [Google Scholar]
  16. China B., Jacquemin E., Devrin A. C., Pirson V., Mainil J.. 1999b; Heterogeneity of the eae genes in attaching/effacing Escherichia coli from cattle: comparison with human strains. Res Microbiol150:323–332[CrossRef]
    [Google Scholar]
  17. Cid D., Blanco M., Blanco J. E., Ruiz Santa Quiteria J. A., Blanco J., De la Fuente R.. 1996; Serogroups, toxins and antibiotic resistance of Escherichia coli strains isolated from diarrhoeic goat kids in Spain. Vet Microbiol53:349–354[CrossRef]
    [Google Scholar]
  18. Donnenberg M. S., Yu J., Kaper J. B.. 1993; A second chromosomal gene necessary for intimate attachment of enteropathogenic Escherichia coli to epithelial cells. J Bacteriol175:4670–4680
    [Google Scholar]
  19. Drolet R., Fairbrother J. M., Vaillancourt D.. 1994; Attaching and effacing Escherichia coli in a goat with diarrhoea. Can Vet J35:122–123
    [Google Scholar]
  20. Duhamel G. E., Moxley R. A., Maddox C. W., Erickson E. D.. 1992; Enteric infection of a goat with enterohemorrhagic Escherichia coli (O103: H2). J Vet Diagn Invest4:197–200[CrossRef]
    [Google Scholar]
  21. Ebel F., Deibel C., Kresse A. U., Guzman C. A., Chakraborty T.. 1996; Temperature- and medium-dependent secretion of proteins by shiga toxin-producing Escherichia coli . Infect Immun64:4472–4479
    [Google Scholar]
  22. Elliot S. J., Wainwright L. A., McDaniel T. K., Jarvis K. G., Deng Y. K., Lai L. C., McNamara B. P., Donnenberg M. S., Kaper J. B.. 1998; The complete sequence of the locus of enterocyte effacement (LEE) from enteropathogenic Escherichia coli E2348/69. Mol Microbiol28:1–4
    [Google Scholar]
  23. Fischer J., Maddox C., Moxley R., Kinden D., Miller M.. 1994; Pathogenicity of a bovine attaching effacing Escherichia coli isolated lacking Shiga-like toxins. Am J Vet Res55:991–999
    [Google Scholar]
  24. Foubister V., Rossenshine I., Donnenberg M. S., Finlay B. B.. 1994; The eaeB of enteropathogenic Escherichia coli is necessary for signal transduction in epithelial cells. Infect Immun62:3038–3040
    [Google Scholar]
  25. Frankel G., Candy D. C. A., Everest P., Dougan G.. 1994; Characterization of the C-terminal domains of intimin-like proteins of enteropathogenic and enterohemorrhagic Escherichia coli , Citrobacter freundii , and Hafnia alvei . Infect Immun62:1835–1842
    [Google Scholar]
  26. Frankel G., Philips A. D., Rosenshine I., Dougan G., Kaper J. B., Knutton S.. 1998; Enteropathogenic and enterohaemorrhagic Escherichia coli : more subversive elements. Mol Microbiol30:911–921[CrossRef]
    [Google Scholar]
  27. Gannon V. P. J., Rashed M., King R. K., Thomas E. J. G.. 1993; Detection and characterization of the eae gene of shiga-like toxin producing Escherichia coli using polymerase chain reaction. J Clin Microbiol31:1268–1274
    [Google Scholar]
  28. Griffin P. M., Tauxe R. V.. 1991; The epidemiology of infections caused by Escherichia coli O157: H7, other enterohemorrhagic E. coli , and the associated hemolytic uremic syndrome. Epidemiol Rev13:60–98
    [Google Scholar]
  29. Higgins D. G., Sharp P. M.. 1988; clustal: a package for performing multiple sequence alignments on a microcomputer. Gene73:237–244[CrossRef]
    [Google Scholar]
  30. Janke B. H., Francis D. H., Collins J. E., Libal M. C., Zeman K. H., Johnson D. D.. 1989; Attaching and effacing Escherichia coli infections in calves, pigs, lambs, and dogs. J Vet Diagn Invest1:6–11[CrossRef]
    [Google Scholar]
  31. Jerse A. E., Yu J., Tall B. D., Kaper J. B.. 1990; A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci USA87:7839–7843[CrossRef]
    [Google Scholar]
  32. Kudva I. T., Hatfield P. G., Hovde C. J.. 1996; Escherichia coli O157: H7 in microbial flora of sheep. J Clin Microbiol34:431–433
    [Google Scholar]
  33. McDaniel T. K., Kaper J. B.. 1997; A cloned pathogenicity island from enteropathogenic Escherichia coli confers the attaching and effacing phenotype on E. coli K-12. Mol Microbiol23:399–407[CrossRef]
    [Google Scholar]
  34. McDaniel T. K., Jarvis K. G., Donnenberg M. S., Kaper J. B.. 1995; A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. Proc Natl Acad Sci USA92:1664–1668[CrossRef]
    [Google Scholar]
  35. McGraw E. A., Li J., Selander R. K., Whittam T. S.. 1999; Molecular evolution and mosaic structure of α, β, and γ intimins of pathogenic Escherichia coli . Mol Biol Evol16:12–22[CrossRef]
    [Google Scholar]
  36. Mainil J. G., Jacquemin E. R., Kaeckenbeeck A. E., Pohl P. H.. 1993; Association between the effacing ( eae ) gene and the Shiga-like toxin encoding genes in Escherichia coli isolated from cattle. Am J Vet Res54:1064–1068
    [Google Scholar]
  37. Milon A., Oswald E., De Rycke J.. 1999; Rabbit EPEC: a model for the study of enteropathogenic Escherichia coli . Vet Res30:203–219
    [Google Scholar]
  38. Moon H. W., Whipp S. C., Argenzio R. A., Levine M. M., Gianella R. A.. 1983; Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines. Infect Immun39:1340–1351
    [Google Scholar]
  39. Moxley R. A., Francis D. H.. 1986; Natural and experimental infection with an attaching and effacing strain of Escherichia coli in calves. Infect Immun53:339–346
    [Google Scholar]
  40. Nataro J. P., Kaper J. B.. 1998; Diarrhoeagenic Escherichia coli . Clin Microbiol Rev11:142–201
    [Google Scholar]
  41. Oswald E., Schmidt H., Morabito S., Karch H., Marches O., Caprioli A.. 2000; Typing of intimin genes in human and animal enterohemorrhagic and enteropathogenic Escherichia coli : characterization of a new intimin variant. Infect Immun68:64–71[CrossRef]
    [Google Scholar]
  42. Perna N. T., Mayhew G. F., Posfai G., Elliott S., Donnenberg M. S., Kaper J. B., Blattner F. R.. 1998; Molecular evolution of a pathogenicicty island from enterohemorrhagic Escherichia coli O157: H7. Infect Immun66:3810–3817
    [Google Scholar]
  43. Rios M., Prado V., Trucksis M., Arellano C., Borie C., Alexandre M., Fica A., Levine M. M.. 1999; Clonal diversity of Chilean isolates of enterohemorrhagic Escherichia coli from patients with hemolytic-uremic syndrome, asymptomatic subjects, animal reservoirs, and food products. J Clin Microbiol37:778–780
    [Google Scholar]
  44. Schauer D. B., Falkow S.. 1993; Attaching and effacing locus of a Citrobacter freundii biotype 4280 that causes transmissible murine colonic hyperplasia. Infect Immun61:2486–2492
    [Google Scholar]
  45. Smith C. L., Cantor C. R.. 1987; Purification, specific fragmentation, and separation of large DNA molecules. Methods Enzymol55:449–467
    [Google Scholar]
  46. Tenover F. C., Arbeit R. D., Goering R. V., Mickelsen P. A., Murray B. E., Persing D. H., Swaminathan B.. 1996; Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis, criteria for bacterial strain typing. J Clin Microbiol33:2233–2239
    [Google Scholar]
  47. Voss E., Paton A. W., Manning P. A., Paton J. C.. 1998; Molecular analysis of shiga toxigenic Escherichia coli O111: H– proteins which react with sera from patients with hemolytic-uremic syndrome. Infect Immun66:1467–1472
    [Google Scholar]
  48. Wells J. G., Shipman L. D., Greene K. D.. 10 other authors 1991; Isolation of Escherichia coli serotype O157: H7 and other Shiga-like-toxin-producing E. coli from dairy cattle. J Clin Microbiol29:985–989
    [Google Scholar]
  49. Whitman T. S., Wolfe M. L., Wachsmuth I. K., Ørskov F., Ørskov I., Wilson R. A.. 1993; Clonal relationships among Escherichia coli strains that cause hemorrhagic colitis and infantile diarrhoea. Infect Immun61:16–19
    [Google Scholar]
  50. Wieler L. H., Schwanitz A., Vieler E., Busse B., Steinruck H., Kaper J. B., Baljer G.. 1998; Virulence properties of Shiga toxin-producing Escherichia coli (STEC) strains of serogroup O118, a major group of STEC pathogens in calves. J Clin Microbiol36:1604–1607
    [Google Scholar]
  51. Yu J., Kaper J. B.. 1992; Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157: H7. Mol Microbiol6:411–417[CrossRef]
    [Google Scholar]
  52. Zhu C., Harel J., Jacques M., Desautels C., Donnenberg M. S., Beaudry M., Fairbrother J. M.. 1994; Virulence properties of attaching-effacing activity of Escherichia coli O45 from swine postweaning diarrhoea. Infect Immun62:4153–4159
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-8-2341
Loading
/content/journal/micro/10.1099/00221287-147-8-2341
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error