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Volume 47, Issue 9

Enteropathogenicity markers in isolated from infants with acute diarrhoea and healthy controls in Rio de Janeiro, Brazil Free

Abstract

Faeces from urban children > 2 years old with acute diarrhoeal illness and from non-diarrhoeal infants (controls) were examined for and other enteropathogens. A total of 990 isolates from 100 patients and 50 controls was tested for enteropathogenic (EPEC) serotype (O:H), adherence to HEp-2 cells after incubation for 3 and 6 h, fluorescent actin staining (FAS), DNA hybridisation with EAF, , STh, STp and EAggEC probes and production of heat-labile enterotoxin (LT) and verocytotoxin (VT) with Y1 and Vero cells. EPEC were the most prevalent enteropathogens in patients (32.7%; and 14% in controls). Enteroinvasive (EIEC) and Vero cytotoxin-producing (VTEC) were not detected. The rate of isolation of enterotoxigenic (ETEC) was identical in both groups. Among the EPEC isolates the prevalent serotypes were O111:H2, O55:NM and O119:H6. Localised adherence (LA) was found significantly more frequently in isolates from patients (19.6%) than controls (2.1%). All LA-positive EPEC isolates were FAS+ and +, but only 75.2% of them hybridised with the EAF probe. Diffusely adhering (DAEC) and enteroaggregative (EAggEC) were found with equal frequency in patients and controls. Twenty-seven isolates were negative for EAF but positive for and FAS and produced LA in 6-h adherence tests. These EAF-/+ strains were the only putative enteropathogen identified in seven patients and were not found in controls. The ability of these strains to elicit ultrastructural cell alterations and cell-signalling events was evaluated in Caco-2 cells (human colon carcinoma cell line) by the gentamicin invasion assay and by transmission electron microscopy. The numbers of intracellular bacteria in cell invasion tests varied from 0.4% to 1.6% of the cell-associated bacteria after a 6-h incubation period. Tyrosine phosphorylation of host cell proteins was assessed in HEp-2 cells by immunofluorescence microscopy and all strains gave positive results. EAF-/+ strains express most of the virulence properties found among true EPEC strains and can be a relevant cause of infant diarrhoea in developing countries.

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© 1998 The Pathological Society of Great Britain and Ireland
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1998-09-01
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References

  1. Levine M. M. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis 1987; 155:377–389
     [Google Scholar]
  2. Katouly M., Jaafari A., Farhoudi-Moghaddam A. A., Ketabi G. R. Aetiological studies of diarrhoeal diseases in infants and young children in Iran. J Trop Med Hyg 1990; 93:22–21
     [Google Scholar]
  3. Gomes T. A. T., Rassi Y., MacDonald K. L. Enteropathogens associated with acute diarrheal disease in urban infants in Sao Paulo, Brazil. J Infect Dis 1991; 164:131–137
     [Google Scholar]
  4. Clausen C. R., Christie D. L. Chronic diarrhea in infants caused by adherent enteropathogenic Escherichia coli. J Pediatr 1982; 100:358–361
     [Google Scholar]
  5. Scaletsky I. C. A., Silva M. L. M., Trabulsi L. R. Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells. Infect Immun 1984; 45:534–536
     [Google Scholar]
  6. Nataro J. P., Kaper J. B., Robins-Browne R., Prado V., Vial P., Levine M. M. Patterns of adherence of diarrheagenic Escherichia coli to HEp-2 cells. Paediatr Infect Dis J 1987; 6:829–831
     [Google Scholar]
  7. Scaletsky I. C. A., Silva M. L. M., Toledo M. R. F., Davis B. R., Blake P. A., Trabulsi L. R. Correlation between adherence to HeLa cells and serogroups, serotypes, and bioserotypes of Escherichia coli. Infect Immun 1985; 49:528–532
     [Google Scholar]
  8. Levine M. M., Prado V., Robins-Browne R. Use of DNA probes and HEp-2 cell adherence assay to detect diarrheagenic Escherichia coli. J Infect Dis 1988; 158:224–228
     [Google Scholar]
  9. Gomes T. A. T., Blake P. A., Trabulsi L. R. Prevalence of Escherichia coli strains with localized, diffuse, and aggregative adherence to HeLa cells in infants with diarrhea and matched controls. J Clin Microbiol 1989; 27:266–269
     [Google Scholar]
  10. Vial P. A., Robins-Browne R. M., Lior H. Characterization of enteroadherent-aggregative Escherichia coli, a putative agent of diarrheal disease. J Infect Dis 1988; 158:70–79
     [Google Scholar]
  11. Bhan M. K., Raj P., Levine M. M. Enteroaggregative Escherichia coli associated with persistent diarrhea in a cohort of rural children in India. J Infect Dis 1989; 159:1061–1064
     [Google Scholar]
  12. Scotland S. M., Smith H. R., Said B., Willshaw G. A., Cheasty T., Rowe B. Identification of enteropathogenic Escherichia coli isolated in Britain as enteroaggregative or as members of a subclass of attaching-and-effacing E. coli not hybridising with the EPEC adherence-factor probe. J Med Microbiol 1991; 35:278–283
     [Google Scholar]
  13. Cravioto A., Tello A., Navarro A. Association of Escherichia coli HEp-2 adherence patterns with type and duration of diarrhoea. Lancet 1991; 337:262–264
     [Google Scholar]
  14. Girón J. A., Jones T., Millán-Velasco F. Diffuse-adhering Escherichia coli (DAEC) as a putative cause of diarrhea in Mayan children in Mexico. J Infect Dis 1991; 163:507–513
     [Google Scholar]
  15. Echeverria P., Serichantalerg O., Changechawalit S. Tissue culture-adherent Escherichia coli in infantile diarrhea. J Infect Dis 1992; 165:141–143
     [Google Scholar]
  16. Baldini M. M., Kaper J. B., Levine M. M., Candy D. C. A., Moon H. W. Plasmid-mediated adhesion in enteropathogenic Escherichia coli. J Pediatr Gastroenterol Nutr 1983; 2:534–538
     [Google Scholar]
  17. Baldini M. M., Nataro J. P., Kaper J. B. Localization of a determinant for HEp-2 adherence by enteropathogenic Escherichia coli. Infect Immun 1986; 52:334–336
     [Google Scholar]
  18. 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; 152:560–565
     [Google Scholar]
  19. Moon H. W., Whipp S. C., Argenzio R. A., Levine M. M., Giannella R. A. Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines. Infect Immun 1983; 41:1340–1351
     [Google Scholar]
  20. 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; 57:1290–1298
     [Google Scholar]
  21. Andrade J., R C., de Santa Rosa M. R. Attachment and intracellular penetration of classic enteropathogenic Escherichia coli into HEp-2 cells. Rev Microbiol (Sao Paulo) 1986; 17:53–57
     [Google Scholar]
  22. Andrade J. R. C., da Veiga V. F., de Santa Rosa M. R., Suassuna I. An endocytic process in HEp-2 cells induced by enteropathogenic Escherichia coli. J Med Microbiol 1989; 28:49–57
     [Google Scholar]
  23. Donnenberg M. S., Donohue-Rolfe A., Keusch G. T. Epithelial cell invasion: an overlooked property of enteropathogenic Escherichia coli (EPEC) associated with the EPEC adherence factor. J Infect Dis 1989; 160:452–459
     [Google Scholar]
  24. Donnenberg M. S., Donohue-Rolfe A., Keusch G. T. A comparison of HEp-2 cell invasion by enteropathogenic and enteroinvasive Escherichia coli. FEMS Microbiol Lett 1990; 69:83–86
     [Google Scholar]
  25. Rosenshine I., Donnenberg M. S., Kaper J. B., Finlay B. B. Signal transduction between enteropathogenic Escherichia coli (EPEC) and epithelial cells: EPEC induces tyrosine phosphorylation of host cell proteins to initiate cytoeskeletal rearrangement and bacterial uptake. EMBO J 1992; 11:3551–3560
     [Google Scholar]
  26. McDaniel T. K., Jarvis K. G., Donnenberg M.S., Kaper J. B. A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. Proc Natl Acad Sci USA 1995; 92:1664–1668
     [Google Scholar]
  27. 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 USA 1990; 87:7839–7843
     [Google Scholar]
  28. Edwards P. R., Ewing W. H. (eds) Identification of Enterobacteriaceae 3rd edn Minneapolis: Burgess Publishing; 1972
     [Google Scholar]
  29. Hausler W. J., Lennette E. H., Balows A., Shadomy H. J. (eds) Manual of clinical microbiology 5th edn Washington, DC: ASM Press; 1991
     [Google Scholar]
  30. Séreny B. Experimental Shigella keratoconjunctivitis. Acta Microbiol Acad Sci Hung 1955; 2:293–296
     [Google Scholar]
  31. Smith H. R., Scotland S. M. Vero cytotoxin-producing strains of Escherichia coli. J Med Microbiol 1988; 26:77–85
     [Google Scholar]
  32. Sack D. A., Sack R. B. Test for enterotoxigenic Escherichia coli using Y-l adrenal cells in miniculture. Infect Immun 1975; 11:334–336
     [Google Scholar]
  33. Cravioto A., Gross R. J., Scotland S. M., Rowe B. An adhesive factor found in strains of Escherichia coli belonging to the traditional infantile enteropathogenic serotypes. Curr Microbiol 1979; 3:98–99
     [Google Scholar]
  34. Maas R. An improved colony hybridization method with significant sensitivity for detection of single genes. Plasmid 1983; 10:296–298
     [Google Scholar]
  35. Harford N., De Wilde M., Cabezon T. Cloning of two distinct but related ST enterotoxin genes from porcine and human strains of E. coli. In Levy S. B., Clowes R. C., Koening E. L. (eds) Molecular biology, pathogenicity, and ecology of bacterial plasmids New York: Plenum Press; 1981611
     [Google Scholar]
  36. Baudry B., Savarino S. J., Vial P., Kaper J. B., Levine M. M. A sensitive and specific DNA probe to identify enteroaggregative Escherichia coli, a recently discovered diarrheal pathogen. J Infect Dis 1990; 161:1249–1251
     [Google Scholar]
  37. Rousset M. The human colon carcinoma cell lines HT-29 and Caco-2: two in vitro models for the study of intestinal differentiation. Biochimie 1986; 68:1035–1040
     [Google Scholar]
  38. Robins-Browne R. M., Bennett-Wood V. Quantitative assessment of the ability of Escherichia coli to invade cultured animal cells. Microb Pathog 1992; 12:159–164
     [Google Scholar]
  39. Rosenshine I., Duronio V., Finlay B. B. Tyrosine protein kinase inhibitors block invasin-promoted bacterial uptake by epithelial cells. Infect Immun 1992; 60:2211–2217
     [Google Scholar]
  40. Toledo M. R. F., Alvariza Mdo C. B., Murahovschi J., Ramos S. R. T. S., Trabulsi L. R. Enteropathogenic Escherichia coli serotypes and endemic diarrhea in infants. Infect Immun 1983; 39:586–589
     [Google Scholar]
  41. Gomes T. A. T., Vieira M. A. M., Wachsmuth I. K., Blake P. A., Trabulsi L. R. Serotype-specific prevalence of Escherichia coli strains with EPEC adherence factor genes in infants with and without diarrhea in Sao Paulo, Brazil. J Infect Dis 1989; 160:131–135
     [Google Scholar]
  42. Magalhães M., Amorim R. J. M., Takeda Y., Tsukamoto T., Antas M. G., Tateno S. Localized, diffuse, and aggregative-adhering Escherichia coli from infants with acute diarrhea and matched-controls. Mem Inst Oswaldo Cruz 1992; 87:93–97
     [Google Scholar]
  43. Albert M. J., Faruque S. M., Faruque A. S. G. Controlled study of Escherichia coli diarrheal infections in Bangladeshi children. J Clin Microbiol 1995; 33:973–977
     [Google Scholar]
  44. Wolk M., Ohad E., Shafran R. Epidemiological aspects of enterotoxigenic Escherichia coli diarrhoea in infants in the Jerusalem area. Public Health Rev 1995; 23:25–33
     [Google Scholar]
  45. Black R. E., Brown K. H. N., Becker S., Abdul A. R. M. A., Huq I. Longitudinal studies of infectious diseases and physical growth of children in rural Bangladesh. II. Incidence of diarrhea and association with known pathogens. Am J Epidemiol 1982; 115:315–324
     [Google Scholar]
  46. Giraldi R., Guth B. E. C., Trabulsi L. R. Production of shiga-like toxin among Escherichia coli strains and other bacteria from diarrhea in Sao Paulo, Brazil. J Clin Microbiol 1990; 28:1460–1462
     [Google Scholar]
  47. Toledo M. R. F., Trabulsi L. R. Frequency of enteroinvasive Escherichia coli in children with diarrhea and healthy controls in Sao Paulo, SP, Brazil. Rev Microbiol (Sao Paulo) 1990; 21:1–4
     [Google Scholar]
  48. Echeverria P., Orskov F., Orskov I. Attaching and effacing enteropathogenic Escherichia coli as a cause of infantile diarrhea in Bangkok. J Infect Dis 1991; 164:550–554
     [Google Scholar]
  49. Nataro J. P., Scaletsky I. C. A., Kaper J. B., Levine M. M., Trabulsi L. R. Plasmid-mediated factors confering diffuse and localized adherence of enteropathogenic Escherichia coli. Infect Immun 1985; 48:378–383
     [Google Scholar]
  50. Scotland S. M., Willshaw G. A., Smith H. R., Gross R. J., Rowe B. Adhesion to cells in culture and plasmid profiles of enteropathogenic Escherichia coli isolated from outbreaks and sporadic cases of infant diarrhoea. J Infect 1989; 19:237–249
     [Google Scholar]
  51. Moyenuddin M., Wachsmuth I. K., Moseley S. L., Bopp C. A., Blake P. A. Serotype, antimicrobial resistance, and adherence properties of Escherichia coli strains associated with outbreaks of diarrheal illness in children in the United States. J Clin Microbiol 1989; 27:2234–2239
     [Google Scholar]
  52. Knutton S., Phillips A. D., Smith H. R. Screening for enteropathogenic Escherichia coli in infants with diarrhea by the fluorescent-actin staining test. Infect Immun 1991; 59:365–371
     [Google Scholar]
  53. Jallat C., Livrelli V., Darfeuille-Michaud A., Rich C., Joly B. Escherichia coli strains involved in diarrhea in France: high prevalence and heterogenicity of diffusely adhering strains. J Clin Microbiol 1993; 31:2031–2037
     [Google Scholar]
  54. Scotland S. M., Smith H. R., Cheasty T. Use of gene probes and adhesion tests to characterise Escherichia coli belonging to enteropathogenic serogroups isolated in the United Kingdom. J Med Microbiol 1996; 44:438–443
     [Google Scholar]
  55. Kaper J. B. Defining EPEC. Rev Microbiol (Sao Paulo) 1996; 27: (Suppl 1) 130–133
     [Google Scholar]
  56. Scotland S. M., Willshaw G. A., Smith H. R., Said B., Stokes N., Rowe B. Virulence properties of Escherichia coli strains belonging to serogroups O26, O55, O111 and O128 isolated in the United Kingdom in 1991 from patients with diarrhoea. Epidemiol Infect 1993; 111:429–438
     [Google Scholar]
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Enteropathogenicity markers in Escherichia coli isolated from infants with acute diarrhoea and healthy controls in Rio de Janeiro, Brazil
J. Med. Microbiol. 47, 781 (1998); https://doi.org/10.1099/00222615-47-9-781
/content/journal/jmm/10.1099/00222615-47-9-781
/content/journal/jmm/10.1099/00222615-47-9-781
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