European Journal of Epidemiology

, Volume 12, Issue 1, pp 13–19 | Cite as

Prevalence and characteristics of human and bovine verotoxigenic Escherichia coli strains isolated in Galicia (north-western Spain)

  • M. Blanco
  • J. E. Blanco
  • J. Blanco
  • E. A. Gonzalez
  • M. P. Alonso
  • H. Maas
  • W. H. Jansen


An epidemiological study was carried out to determine the incidence and the serotypes of verotoxigenic Escherichia coli (VTEC) that cause infections in Galicia (north-western Spain). Although, VTEC strains were isolated from 55 (14%) of the 387 calves sampled and the majority of bovine VTEC strains belonged to serotypes (026:H11 or H−, 091:H21, 0103:H2, 0105:H18, 0111:H−, 0113:H21, 0126:H−, 0128:H− and 0157:H7 or H−) previously associated with human haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS) in other countries, VTEC are not a common cause of human infections in Spain. Thus, VTEC (026:H11 and 086:H10) were isolated from only 3 (0.6%) of the 482 children with diarrhoea investigated. We examined the 69 (3 humans and 66 bovines) VTEC strains that were initially isolated as E. coli producing a toxin cytotoxic to Vero and HeLa cells by polymerase chain reaction (PCR) using specific primers for VT1, VT2 and eae genes. PCR showed that 38 (55%) of VTEC strains carried VT1 genes, 18 (26%) possessed VT2 genes, and 10 (14%) carried both VT1 and VT2 genes. Three (one human and two bovine) strains which were formerly VTEC had lost the ability to produce verotoxins upon subculture and became negative for VT 1 and VT2 by PCR. In total 35 (51%) of 69 VTEC strains, including the two human VT1+ strains of serotype 026:H11, were positive for eae sequences when tested by PCR. Presence of the eae gene was significantly more frequent (100%; 21/21) among VTEC strains with serotypes (026:H11, 0111:H−, 0157:H−and 0157:H7) considered as enterohaemorrhagic E. coli (EHEC) than among VTEC strains with non-EHEC serotypes (29%; 14/48) (p < 0.001). Results obtained in this study indicate that cattle may be an important source of VTEC involved in human disease. However, severe clinical syndromes caused by VTEC, such as HC and HUS, are uncommon in Spain, in comparison with North America and the UK. In any case, VTEC disease can appear on the scene very suddenly, as occurred in the UK and North America in the 1980s.

Key words

Escherichia coli Pathogenicity Shiga-like toxins Verotoxins 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Blanco J, Blanco M, Blanco JE, Alonso MP, Escribano A. Patogénesis, epidemiología y diagnóstico microbiológico de las infecciones producidas por Escherichia coli enterohemorrágicos productores de verotoxinas. Enferm Infecc Microbiol Clin 1993; 11: 324–334.Google Scholar
  2. 2.
    Griffin PM, Tauxe RV. The epidemiology of infections caused by Escherichia coli 0157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev 1991; 13: 60–98.Google Scholar
  3. 3.
    Karmali MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev 1989; 2: 15–38.Google Scholar
  4. 4.
    Levine MM. Escherichia coli that cause diarrhea: Enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis 1987; 155: 377–389.Google Scholar
  5. 5.
    Smith HR, Scotland SM. Vero cytotoxin-producing strains of Escherichia coli. J Med Microbiol 1988; 26: 77–85.Google Scholar
  6. 6.
    Carter AO, Borczyk AA, Carlson JAK, Harvey B, Hockin JC, Karmali MA, Krishnan C, Kom DA, Lior H. A severe outbreak of Escherichia coli 0157:H7-associated hemorrhagic colitis in a nursing home. N Engl J Med 1987; 317: 1496–1500.Google Scholar
  7. 7.
    Barret TJ, Lior H, Green JH, Khakhria R, Wells JG, Bell BP, Greene KD, Lewis J, Griffin PM. Laboratory investigation of a multistate food-borne outbreak of Escherichia coli 0157:H7 by using pulsed-field gel electrophoresis and phage typing. J Clin Microbiol 1994; 32: 3013–3017.Google Scholar
  8. 8.
    Rowe PC, Orrbine E, Lior H, Wells GA, McLaine PN, CPKDRC co-investigators. A prospective study of exposure to verotoxin-producing Escherichia coli among Canadian children with haemolytic uraemic syndrome. Epidemiol Infect 1993; 110: 1–7.Google Scholar
  9. 9.
    Salmon RL, Farrell ID, Hutchison JGP, Coleman DJ, Gross RJ, Fry NK, Rowe B, Palmer SR. A christening party outbreak of haemorrhagic colitis and haemolytic uraemic syndrome associated with Escherichia coli 0157:H7. Epidemiol Infect 1989; 103: 249–254.Google Scholar
  10. 10.
    Beutin L, Aleksic S, Zimmermann S, Gleier K Virulence factors and phenotypical traits of verotoxigenic strains of Escherichia coli isolated from human patients in Germany. Med Microbiol Immunol 1994; 183: 13–21.Google Scholar
  11. 11.
    Neill MA. Pathogenesis of Escherichia coli 0157:H7 infection. Curr Opin Infect Dis 1994; 7: 295–303.Google Scholar
  12. 12.
    Barret TJ, Kaper JB, Jerse AE, Wachsmuth IK. Virulence factors in Shiga-like toxin-producing Escherichia coli isolated from humans and cattle. J Infect Dis 1992; 165: 979–980.Google Scholar
  13. 13.
    Gannon VPJ, Rashed M, King RK, Golsteyn Thomas EJ. Detection and characterization of the eae gene of shiga-like toxin-producing Escherichia coli using polymerase chain reaction. J Clin Microbiol 1993; 31: 1268–1274.Google Scholar
  14. 14.
    Marques LRM, Peiris JSM, O'Brien AD. Escherichia coli strains isolated from pigs with edema disease produce a variant of Shiga-like toxin II. FEMS Microbiol Lett 1987; 44: 33–38.Google Scholar
  15. 15.
    Olsvik Ø, Strockbine NA. PCR detection of heat-stable, heat-labile, and Shiga-like toxin genes in Escherichia coli. In: Persing DH, Smith TF, Tenover FC and White TJ (eds), Diagnostic molecular microbiology. Principles and applications. Washington, DC: American Society for Microbiology, 1993: 271–276.Google Scholar
  16. 16.
    Guinee PAM, Jansen HW, Wadström T, Sellwood R. Escherichia coli associated with neonatal diarrhoea in piglets and calves. In: De Leeww PW, Guinee PAM (eds), Current topics in veterinary and animal Science, Vol. 13. The Hague: Martinus Nijhoff, 1981: 126–162.Google Scholar
  17. 17.
    Blanco M, Blanco J, Blanco JE, Ramos J. Enterotoxigenic, verotoxigenic and necrotoxigenic Escherichia coli isolated from cattle in Spain. Am J Vet Res 1993; 54: 1446–1451.Google Scholar
  18. 18.
    Blanco M, Blanco J, Blanco JE, González EA, Tardelli TA, Zerbini F, Yano T, Pestana de Castro AF. Genes coding for Shiga-like toxins in bovine verotoxin-producing E. coli (VTEC) strains belonging to differents O:K:H serotypes. Vet Microbiol 1994; 42: 105–110.Google Scholar
  19. 19.
    Blanco J, González EA, Garcia S, Blanco M, Regueiro B, Bernárdez I. Production of toxins by Escherichia coli strains isolated from calves with diarrhoea in Galicia (north-western Spain). Vet Microbiol 1988; 18: 297–311.Google Scholar
  20. 20.
    Mohammad A, Peiris JSM, Wijewanta EA. Serotypes of verocytoxigenic Escherichia coli isolated from cattle and buffalo calf diarrhoea. FEMS Microbiol Lett 1986; 35: 261–265.Google Scholar
  21. 21.
    Sherwood D, Snodgrass DR, O'Brien AD. Shiga-like toxin production from Escherichia coli associated with calf diarrhoea. Vet Rec 1985; 116: 217–218.Google Scholar
  22. 22.
    Wells JG, Shipman LD, Greene KD, Sowers EG, Green JH, Cameron DN, Downes FP, Mantin ML, Griffin PM, Ostroff SM, Potter ME, Tauxe RV, Wachsmuth IK. Isolation of Escherichia coli serotype 0157:H7 and other shiga-like-toxin-producing Escherichia coli from dairy cattle. J Clin Microbiol 1991; 29: 985–989.Google Scholar
  23. 23.
    Willshaw GA, Cheasty T, Jiggle B, Rowe B, Gibbons D, Hutchinson DN. Vero cytotoxin-producing Escherichia coli in a herd of dairy cattle. Vet Rec 1993; 132: 96.Google Scholar
  24. 24.
    Wilson JB, McEwen SA, Clarke RC, Leslie KE, Wilson RA, Waltner-Toews D, Gyles CL. Distribution and characteristics of verocytotoxigenic Escherichia coli isolated from Ontario dairy cattle. Epidemiol Infect 1992; 108: 423–439.Google Scholar
  25. 25.
    Pohl P, Daube G, Lintermans P, Kaeckenbeeck A, Mainil J. Description de 70 souches d'Escherichia coli d'origine bovine possedant les genes des verotoxines. Ann Med Vet 1991; 135: 267–272.Google Scholar
  26. 26.
    Hales BA, Fletcher JN, Ridha G, Batt RM, Hart CA, Saunders JR. Incidence of common DNA sequences in bovine and porcine Escherichia coli strains causing diarrhoea. Res Vet Sci 1991; 50: 355–357.Google Scholar
  27. 27.
    Mohammad A, Peiris JSM, Scotland SM, Willshaw GA, Smith HR, Cheasty T. A longitudinal study of Vero cytotoxin producing Escherichia coli in cattle calves in Sri Lanka. Epidemiol Infect 1993; 110: 197–298.Google Scholar
  28. 28.
    Dom CR, Francis DH, Angrick EJ, Willgohs JA, Wilson RA, Collis JE, Janke BH, Shawd SJ. Characteristics of Vero cytotoxin producing Escherichia coli associated with intestinal colonization and diarrhea in calves. Vet Microbiol 1993; 36: 149–159.Google Scholar
  29. 29.
    Montenegro MA, BHlte M, Trumpf T, Aleksia S, Reuter G, Bulling E, Helmuth R. Detection and characterization of fecal verotoxin-producing Escherichia coli from healthy cattle. J Clin Microbiol 1990; 28: 1417–1421.Google Scholar
  30. 30.
    Beutin L, Geier D, SteinrHck H, Zimmermann S, Scheutz F. Prevalence and some properties of verotoxin (Shiga-like toxin)-producing Escherichia coli in seven different species of healthy domestic animals. J Clin Microbiol 1993; 31: 2483–2488.Google Scholar
  31. 31.
    Tesh VL, Burris JA, Owens JW, Gordon VM, Wadolkowski EA, O'Brien AD, Samuel JE. Comparison of the relative toxicities of Shiga-like toxins type I and II for mice. Infect Immun 1993; 61: 3392–3402.Google Scholar
  32. 32.
    Cahoon FC, Thompson JS. Frequency of Escherichia coli 0157:H7 isolation from stool specimens. Can J Microbiol 1987; 33: 914–915.Google Scholar
  33. 33.
    Edelman R, Karmali MA, Fleming PA. Summary of the international symposium and workshop on infections due to verocytotoxin (Shiga-like toxin) producing Escherichia coli. J Infect Dis 1988; 157: 1102–1104.Google Scholar
  34. 34.
    Sharp JCM, Coia JE, Curnow J, Reilly WJ. Escherichia coli 0157 infections in Scotland. J Med Microbiol 1994; 40: 3–9.Google Scholar
  35. 35.
    Gaztelurrutia L, Barren J, Prats G. Colitis hemorrágica por Escherichia coli 0157: H7 verotoxigénico. Enferm Infecc Microbiol Clin 1990; 8: 320–321.Google Scholar
  36. 36.
    Marne C, Puig A, Jovani A. Investigación de Escherichia coli 0157:H7 en la comarca del Maresme (Barcelona). Enferm Infecc Microbiol Clin 1990; 8: 465–466.Google Scholar
  37. 37.
    Levine MM, Xu JG, Kaper JB, Lior H, Prodo V, Tall B, Nataro J, Karch H, Wachsmuth K. A DNA probe to identify enterohemorrhagic Escherichia coli of 0157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis 1987; 156: 175–182.Google Scholar
  38. 38.
    Jerse AE, Yu J, Tall BD, Karper JB. 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

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • M. Blanco
    • 1
  • J. E. Blanco
    • 1
  • J. Blanco
    • 1
  • E. A. Gonzalez
    • 1
  • M. P. Alonso
    • 2
  • H. Maas
    • 2
  • W. H. Jansen
    • 3
  1. 1.Departamento de Microbioloxía e Parasitoloxía, Facultade de VeterinariaUniversidade de SantiagoLugoSpain
  2. 2.Unidade de MicrobioloxíaComplexo Hospitalario Xeral-CaldeLugoSpain
  3. 3.Laboratory of Bacteriology and Antimicrobial AgentsNational Institute of Public Health and Environmental ProtectionBilthovenThe Netherlands

Personalised recommendations