Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Nov;27(11):2559–2564. doi: 10.1128/jcm.27.11.2559-2564.1989

Close association of verotoxin (Shiga-like toxin) production with enterohemolysin production in strains of Escherichia coli.

L Beutin 1, M A Montenegro 1, I Orskov 1, F Orskov 1, J Prada 1, S Zimmermann 1, R Stephan 1
PMCID: PMC267076  PMID: 2681256

Abstract

Sixty-four verotoxin-producing (VT+) Escherichia coli strains were analyzed for VT1- and VT2-specific DNA sequences and for production of hemolysin. Strains of human origin were of the following serotypes: O157:H7 or H-, O111:H8 or H-, O26:H11, O114:H4, and rough:H7. Strains of serotypes O157:H7, O113:H21, O116:H21, and rough:H- were from cattle, while those of serotype O139:K12:H1 were from pigs. All 64 isolates carried either VT1 or VT2 or both genes. Sixty of the strains (93.8%) were hemolytic (Hly+). The three O139:K12:H1 strains examined produced alpha-hemolysin, as shown by their reaction with the alpha-hemolysin-specific monoclonal antibody h2A and by DNA hybridization with an alpha-hly gene probe. The remaining 57 Hly+ strains (95%) produced a different type of hemolysin (enterohemolysin), which is genetically and serologically unrelated to alpha-hemolysin. The two types of hemolysin are further distinguished by the appearance of the lysis zone on blood agar and by the time interval for the detection of hemolysis. In contrast to alpha-hemolysin, enterohemolysin can be detected only on blood plates containing washed erythrocytes. The frequent association of enterohemolysin with verotoxin production (89%) makes it useful as an epidemiological marker for rapid and simple detection of potential VT+ E. coli.

Full text

PDF
2559

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Achtman M., Mercer A., Kusecek B., Pohl A., Heuzenroeder M., Aaronson W., Sutton A., Silver R. P. Six widespread bacterial clones among Escherichia coli K1 isolates. Infect Immun. 1983 Jan;39(1):315–335. doi: 10.1128/iai.39.1.315-335.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beutin L., Bode L., Richter T., Peltre G., Stephan R. Rapid visual detection of Escherichia coli and Vibrio cholerae Heat-labile enterotoxins by nitrocellulose enzyme-linked immunosorbent assay. J Clin Microbiol. 1984 Mar;19(3):371–375. doi: 10.1128/jcm.19.3.371-375.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beutin L., Montenegro M., Zimmermann S., Stephan R. Characterization of hemolytic strains of Escherichia coli belonging to classical enteropathogenic O-serogroups. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 May;261(3):266–279. doi: 10.1016/s0176-6724(86)80044-x. [DOI] [PubMed] [Google Scholar]
  4. Beutin L., Prada J., Zimmermann S., Stephan R., Orskov I., Orskov F. Enterohemolysin, a new type of hemolysin produced by some strains of enteropathogenic E. coli (EPEC). Zentralbl Bakteriol Mikrobiol Hyg A. 1988 Mar;267(4):576–588. doi: 10.1016/s0176-6724(88)80042-7. [DOI] [PubMed] [Google Scholar]
  5. Bohnert M. G., d'Hauteville H. M., Sansonetti P. J. Detection of enteric pathotypes of Escherichia coli by hybridization using six DNA probes. Ann Inst Pasteur Microbiol. 1988 Mar-Apr;139(2):189–202. [PubMed] [Google Scholar]
  6. Bopp C. A., Greene K. D., Downes F. P., Sowers E. G., Wells J. G., Wachsmuth I. K. Unusual verotoxin-producing Escherichia coli associated with hemorrhagic colitis. J Clin Microbiol. 1987 Aug;25(8):1486–1489. doi: 10.1128/jcm.25.8.1486-1489.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cavalieri S. J., Bohach G. A., Snyder I. S. Escherichia coli alpha-hemolysin: characteristics and probable role in pathogenicity. Microbiol Rev. 1984 Dec;48(4):326–343. doi: 10.1128/mr.48.4.326-343.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crichton P. B., Old D. C. Biotyping of Escherichia coli. J Med Microbiol. 1979 Nov;12(4):473–486. doi: 10.1099/00222615-12-4-473. [DOI] [PubMed] [Google Scholar]
  9. Edelman R., Karmali M. A., Fleming P. A. From the National Institutes of Health. Summary of the International Symposium and Workshop on Infections due to Verocytotoxin (Shiga-like toxin)-producing Escherichia coli. J Infect Dis. 1988 May;157(5):1102–1104. doi: 10.1093/infdis/157.5.1102. [DOI] [PubMed] [Google Scholar]
  10. Evans D. J., Jr, Evans D. G., Höhne C., Noble M. A., Haldane E. V., Lior H., Young L. S. Hemolysin and K antigens in relation to serotype and hemagglutination type of Escherichia coli isolated from extraintestinal infections. J Clin Microbiol. 1981 Jan;13(1):171–178. doi: 10.1128/jcm.13.1.171-178.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farmer J. J., 3rd, Davis B. R. H7 antiserum-sorbitol fermentation medium: a single tube screening medium for detecting Escherichia coli O157:H7 associated with hemorrhagic colitis. J Clin Microbiol. 1985 Oct;22(4):620–625. doi: 10.1128/jcm.22.4.620-625.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hacker J., Schröter G., Schrettenbrunner A., Hughes C., Goebel W. Hemolytic Escherichia coli strains in the human fecal flora as potential urinary pathogens. Zentralbl Bakteriol Mikrobiol Hyg A. 1983 May;254(3):370–378. [PubMed] [Google Scholar]
  13. Haldane D. J., Damm M. A., Anderson J. D. Improved biochemical screening procedure for small clinical laboratories for vero (Shiga-like)-toxin-producing strains of Escherichia coli O157:H7. J Clin Microbiol. 1986 Oct;24(4):652–653. doi: 10.1128/jcm.24.4.652-653.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hugo F., Arvand M., Reichwein J., Mackman N., Holland I. B., Bhakdi S. Identification with monoclonal antibodies of hemolysin produced by clinical isolates of Escherichia coli. J Clin Microbiol. 1987 Jan;25(1):26–30. doi: 10.1128/jcm.25.1.26-30.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Karmali M. A., Petric M., Lim C., Fleming P. C., Arbus G. S., Lior H. The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli. J Infect Dis. 1985 May;151(5):775–782. doi: 10.1093/infdis/151.5.775. [DOI] [PubMed] [Google Scholar]
  16. Krishnan C., Fitzgerald V. A., Dakin S. J., Behme R. J. Laboratory investigation of outbreak of hemorrhagic colitis caused by Escherichia coli O157:H7. J Clin Microbiol. 1987 Jun;25(6):1043–1047. doi: 10.1128/jcm.25.6.1043-1047.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. March S. B., Ratnam S. Sorbitol-MacConkey medium for detection of Escherichia coli O157:H7 associated with hemorrhagic colitis. J Clin Microbiol. 1986 May;23(5):869–872. doi: 10.1128/jcm.23.5.869-872.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Montenegro M. A., Bitter-Suermann D., Timmis J. K., Agüero M. E., Cabello F. C., Sanyal S. C., Timmis K. N. traT gene sequences, serum resistance and pathogenicity-related factors in clinical isolates of Escherichia coli and other gram-negative bacteria. J Gen Microbiol. 1985 Jun;131(6):1511–1521. doi: 10.1099/00221287-131-6-1511. [DOI] [PubMed] [Google Scholar]
  19. Newland J. W., Neill R. J. DNA probes for Shiga-like toxins I and II and for toxin-converting bacteriophages. J Clin Microbiol. 1988 Jul;26(7):1292–1297. doi: 10.1128/jcm.26.7.1292-1297.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orskov F., Orskov I., Villar J. A. Cattle as reservoir of verotoxin-producing Escherichia coli O157:H7. Lancet. 1987 Aug 1;2(8553):276–276. doi: 10.1016/s0140-6736(87)90860-9. [DOI] [PubMed] [Google Scholar]
  21. Pai C. H., Ahmed N., Lior H., Johnson W. M., Sims H. V., Woods D. E. Epidemiology of sporadic diarrhea due to verocytotoxin-producing Escherichia coli: a two-year prospective study. J Infect Dis. 1988 May;157(5):1054–1057. doi: 10.1093/infdis/157.5.1054. [DOI] [PubMed] [Google Scholar]
  22. Riley L. W., Remis R. S., Helgerson S. D., McGee H. B., Wells J. G., Davis B. R., Hebert R. J., Olcott E. S., Johnson L. M., Hargrett N. T. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med. 1983 Mar 24;308(12):681–685. doi: 10.1056/NEJM198303243081203. [DOI] [PubMed] [Google Scholar]
  23. Scotland S. M., Rowe B., Smith H. R., Willshaw G. A., Gross R. J. Vero cytotoxin-producing strains of Escherichia coli from children with haemolytic uraemic syndrome and their detection by specific DNA probes. J Med Microbiol. 1988 Apr;25(4):237–243. doi: 10.1099/00222615-25-4-237. [DOI] [PubMed] [Google Scholar]
  24. Scotland S. M., Smith H. R., Rowe B. Two distinct toxins active on Vero cells from Escherichia coli O157. Lancet. 1985 Oct 19;2(8460):885–886. doi: 10.1016/s0140-6736(85)90146-1. [DOI] [PubMed] [Google Scholar]
  25. Scotland S. M., Willshaw G. A., Smith H. R., Rowe B. Properties of strains of Escherichia coli belonging to serogroup O157 with special reference to production of Vero cytotoxins VT1 and VT2. Epidemiol Infect. 1987 Dec;99(3):613–624. doi: 10.1017/s0950268800066462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smith H. R., Scotland S. M., Willshaw G. A., Wray C., McLaren I. M., Cheasty T., Rowe B. Vero cytotoxin production and presence of VT genes in Escherichia coli strains of animal origin. J Gen Microbiol. 1988 Mar;134(3):829–834. doi: 10.1099/00221287-134-3-829. [DOI] [PubMed] [Google Scholar]
  27. Strockbine N. A., Marques L. R., Holmes R. K., O'Brien A. D. Characterization of monoclonal antibodies against Shiga-like toxin from Escherichia coli. Infect Immun. 1985 Dec;50(3):695–700. doi: 10.1128/iai.50.3.695-700.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Welch R. A., Hull R., Falkow S. Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli. Infect Immun. 1983 Oct;42(1):178–186. doi: 10.1128/iai.42.1.178-186.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Willshaw G. A., Smith H. R., Scotland S. M., Field A. M., Rowe B. Heterogeneity of Escherichia coli phages encoding Vero cytotoxins: comparison of cloned sequences determining VT1 and VT2 and development of specific gene probes. J Gen Microbiol. 1987 May;133(5):1309–1317. doi: 10.1099/00221287-133-5-1309. [DOI] [PubMed] [Google Scholar]
  30. Willshaw G. A., Smith H. R., Scotland S. M., Rowe B. Cloning of genes determining the production of vero cytotoxin by Escherichia coli. J Gen Microbiol. 1985 Nov;131(11):3047–3053. doi: 10.1099/00221287-131-11-3047. [DOI] [PubMed] [Google Scholar]
  31. ØRSKOV F. On the occurrence of E. coli belonging to O-group 26 in cases of infantile diarrhoea and white scours. Acta Pathol Microbiol Scand. 1951;29(4):373–378. [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES