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Canadian Journal of Veterinary Research logoLink to Canadian Journal of Veterinary Research
. 1995 Oct;59(4):265–270.

Detection of toxin genes in Escherichia coli isolated from normal dogs and dogs with diarrhea.

J Hammermueller 1, S Kruth 1, J Prescott 1, C Gyles 1
PMCID: PMC1263780  PMID: 8548687

Abstract

The etiology of acute, nonviral diarrhea in dogs is poorly understood. Enterotoxigenic and verotoxigenic Escherichia coli are causal agents of diarrhea in humans, pigs, and cattle, but the association of these toxigenic E. coli with diarrhea in dogs has not been explored to a significant extent. In this study, DNA hybridization and PCR amplification were used to identify the frequency with which the genes for E. coli enterotoxins (STap, STb, and LTI) and verotoxins (VT1 and VT2) occur in association with diarrhea in dogs. Genes for VT1 (8.9%), VT2 (22.2%), STa (26.7%), and STb (4.4%) were identified in E. coli cultured from feces of 20 of 45 dogs (44.4%) with diarrhea. Genes for VT2, STa, and STb were not identified in feces from normal dogs. Genes for VT1 were observed in similar proportions in fecal samples from diarrheic (8.9%) and normal (12.3%) dogs. Heat labile enterotoxin (LTI) was not detected in fecal samples from either diarrheic or normal dogs. Our results suggest that heat stable enterotoxins and VT2 may be causally associated with diarrhea in dogs. Dogs appear to be able to carry VT1-producing E. coli without showing overt signs of disease.

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Selected References

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  1. Abaas S., Franklin A., Kühn I., Orskov F., Orskov I. Cytotoxin activity on Vero cells among Escherichia coli strains associated with diarrhea in cats. Am J Vet Res. 1989 Aug;50(8):1294–1296. [PubMed] [Google Scholar]
  2. Baljer G., Hinsch F., Mayr B. Klinische Erfahrungen mit der zwingerspezifischen E.-coli Schluckimpfunbei Hunden. Tierarztl Prax. 1990 Feb;18(1):65–68. [PubMed] [Google Scholar]
  3. Brown J. E., Sethabutr O., Jackson M. P., Lolekha S., Echeverria P. Hybridization of Escherichia coli producing Shiga-like toxin I, Shiga-like toxin II, and a variant of Shiga-like toxin II with synthetic oligonucleotide probes. Infect Immun. 1989 Sep;57(9):2811–2814. doi: 10.1128/iai.57.9.2811-2814.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burgess M. N., Bywater R. J., Cowley C. M., Mullan N. A., Newsome P. M. Biological evaluation of a methanol-soluble, heat-stable Escherichia coli enterotoxin in infant mice, pigs, rabbits, and calves. Infect Immun. 1978 Aug;21(2):526–531. doi: 10.1128/iai.21.2.526-531.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collins J. E., Bergeland M. E., Lindeman C. J., Duimstra J. R. Enterococcus (Streptococcus) durans adherence in the small intestine of a diarrheic pup. Vet Pathol. 1988 Sep;25(5):396–398. doi: 10.1177/030098588802500513. [DOI] [PubMed] [Google Scholar]
  6. Dean A. G., Ching Y. C., Williams R. G., Harden L. B. Test for Escherichia coli enterotoxin using infant mice: application in a study of diarrhea in children in Honolulu. J Infect Dis. 1972 Apr;125(4):407–411. doi: 10.1093/infdis/125.4.407. [DOI] [PubMed] [Google Scholar]
  7. Dubreuil J. D., Fairbrother J. M., Lallier R., Larivière S. Production and purification of heat-stable enterotoxin b from a porcine Escherichia coli strain. Infect Immun. 1991 Jan;59(1):198–203. doi: 10.1128/iai.59.1.198-203.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fox J. G., Moore R., Ackerman J. I. Campylobacter jejuni-associated diarrhea in dogs. J Am Vet Med Assoc. 1983 Dec 15;183(12):1430–1433. [PubMed] [Google Scholar]
  9. Gannon V. P., Gyles C. L., Friendship R. W. Characteristics of verotoxigenic Escherichia coli from pigs. Can J Vet Res. 1988 Jul;52(3):331–337. [PMC free article] [PubMed] [Google Scholar]
  10. Georges M. C., Wachsmuth I. K., Meunier D. M., Nebout N., Didier F., Siopathis M. R., Georges A. J. Parasitic, bacterial, and viral enteric pathogens associated with diarrhea in the Central African Republic. J Clin Microbiol. 1984 May;19(5):571–575. doi: 10.1128/jcm.19.5.571-575.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gyles C. L., Barnum D. A. A heat-labile enterotoxin from strains of Eschericha coli enteropathogenic for pigs. J Infect Dis. 1969 Oct;120(4):419–426. doi: 10.1093/infdis/120.4.419. [DOI] [PubMed] [Google Scholar]
  12. Gyles C. L., De Grandis S. A., MacKenzie C., Brunton J. L. Cloning and nucleotide sequence analysis of the genes determining verocytotoxin production in a porcine edema disease isolate of Escherichia coli. Microb Pathog. 1988 Dec;5(6):419–426. doi: 10.1016/0882-4010(88)90003-4. [DOI] [PubMed] [Google Scholar]
  13. Gyles C. L. Escherichia coli cytotoxins and enterotoxins. Can J Microbiol. 1992 Jul;38(7):734–746. doi: 10.1139/m92-120. [DOI] [PubMed] [Google Scholar]
  14. Hammermueller J. D., Krell P. J., Derbyshire J. B., Nagy E. An improved dot-blot method for virus detection in chicken embryo fibroblast cultures. J Virol Methods. 1991 Jan;31(1):47–56. doi: 10.1016/0166-0934(91)90143-n. [DOI] [PubMed] [Google Scholar]
  15. Huang A., de Grandis S., Friesen J., Karmali M., Petric M., Congi R., Brunton J. L. Cloning and expression of the genes specifying Shiga-like toxin production in Escherichia coli H19. J Bacteriol. 1986 May;166(2):375–379. doi: 10.1128/jb.166.2.375-379.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson W. M., Pollard D. R., Lior H., Tyler S. D., Rozee K. R. Differentiation of genes coding for Escherichia coli verotoxin 2 and the verotoxin associated with porcine edema disease (VTe) by the polymerase chain reaction. J Clin Microbiol. 1990 Oct;28(10):2351–2353. doi: 10.1128/jcm.28.10.2351-2353.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Karch H., Meyer T. Single primer pair for amplifying segments of distinct Shiga-like-toxin genes by polymerase chain reaction. J Clin Microbiol. 1989 Dec;27(12):2751–2757. doi: 10.1128/jcm.27.12.2751-2757.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Karmali M. A. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989 Jan;2(1):15–38. doi: 10.1128/cmr.2.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kruth S. A., Prescott J. F., Welch M. K., Brodsky M. H. Nosocomial diarrhea associated with enterotoxigenic Clostridium perfringens infection in dogs. J Am Vet Med Assoc. 1989 Aug 1;195(3):331–334. [PubMed] [Google Scholar]
  20. Lee C. H., Moseley S. L., Moon H. W., Whipp S. C., Gyles C. L., So M. Characterization of the gene encoding heat-stable toxin II and preliminary molecular epidemiological studies of enterotoxigenic Escherichia coli heat-stable toxin II producers. Infect Immun. 1983 Oct;42(1):264–268. doi: 10.1128/iai.42.1.264-268.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Levine M. M. Travellers' diarrhoea: prospects for successful immunoprophylaxis. Scand J Gastroenterol Suppl. 1983;84:121–134. [PubMed] [Google Scholar]
  23. Linggood M. A., Thompson J. M. Verotoxin production among porcine strains of Escherichia coli and its association with oedema disease. J Med Microbiol. 1987 Dec;24(4):359–362. doi: 10.1099/00222615-24-4-359. [DOI] [PubMed] [Google Scholar]
  24. Lopez E. L., Diaz M., Devoto S., Grinstein S., Woloj M., Murray B. E., Rubeglio E., Mendilaharzu F., Turco M., Vasquez M. Evidence of infection with organisms producing Shiga-like toxins in household contacts of children with the hemolytic uremic syndrome. Pediatr Infect Dis J. 1991 Jan;10(1):20–24. doi: 10.1097/00006454-199101000-00005. [DOI] [PubMed] [Google Scholar]
  25. MacLeod D. L., Gyles C. L. Purification and characterization of an Escherichia coli Shiga-like toxin II variant. Infect Immun. 1990 May;58(5):1232–1239. doi: 10.1128/iai.58.5.1232-1239.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mainil J., Daube G., Deprez P., Kaeckenbeeck A., Pohl P. Detection and identification of pathotypes of verocytotoxigenic Escherichia coli isolated from weaned piglets using gene probes for seven E. coli toxins. FEMS Microbiol Lett. 1989 Jun;50(3):345–349. doi: 10.1016/0378-1097(89)90443-6. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Olson P., Hedhammar A., Faris A., Krovacek K., Wadström T. Enterotoxigenic Escherichia coli (ETEC) and Klebsiella pneumoniae isolated from dogs with diarrhoea. Vet Microbiol. 1985 Dec;10(6):577–589. doi: 10.1016/0378-1135(85)90066-5. [DOI] [PubMed] [Google Scholar]
  29. Papageorges M., Higgins R., Gosselin Y. Yersinia enterocolitica in two dogs. J Am Vet Med Assoc. 1983 Mar 15;182(6):618–619. [PubMed] [Google Scholar]
  30. Picken R. N., Mazaitis A. J., Maas W. K., Rey M., Heyneker H. Nucleotide sequence of the gene for heat-stable enterotoxin II of Escherichia coli. Infect Immun. 1983 Oct;42(1):269–275. doi: 10.1128/iai.42.1.269-275.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pollard D. R., Johnson W. M., Lior H., Tyler S. D., Rozee K. R. Differentiation of Shiga toxin and Vero cytotoxin type 1 genes by polymerase chain reaction. J Infect Dis. 1990 Nov;162(5):1195–1198. doi: 10.1093/infdis/162.5.1195. [DOI] [PubMed] [Google Scholar]
  32. Pollard D. R., Johnson W. M., Lior H., Tyler S. D., Rozee K. R. Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction. J Clin Microbiol. 1990 Mar;28(3):540–545. doi: 10.1128/jcm.28.3.540-545.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Read S. C., Clarke R. C., Martin A., De Grandis S. A., Hii J., McEwen S., Gyles C. L. Polymerase chain reaction for detection of verocytotoxigenic Escherichia coli isolated from animal and food sources. Mol Cell Probes. 1992 Apr;6(2):153–161. doi: 10.1016/0890-8508(92)90060-b. [DOI] [PubMed] [Google Scholar]
  34. Read S. C., Gyles C. L., Clarke R. C., Lior H., McEwen S. Prevalence of verocytotoxigenic Escherichia coli in ground beef, pork, and chicken in southwestern Ontario. Epidemiol Infect. 1990 Aug;105(1):11–20. doi: 10.1017/s0950268800047592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sack R. B., Johnson J., Pierce N. F., Keren D. F., Yardley J. H. Challenge of dogs with live enterotoxigenic Escherichia coli and effects of repeated challenges on fluid secretion in jejunal Thiry-Vella loops. J Infect Dis. 1976 Jul;134(1):15–24. doi: 10.1093/infdis/134.1.15. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Smith H. W., Halls S. Studies on Escherichia coli enterotoxin. J Pathol Bacteriol. 1967 Apr;93(2):531–543. doi: 10.1002/path.1700930212. [DOI] [PubMed] [Google Scholar]
  38. Smith H. W., Halls S. The production of oedema disease and diarrhoea in weaned pigs by the oral administration of Escherichia coli: factors that influence the course of the experimental disease. J Med Microbiol. 1968 Aug;1(1):45–59. doi: 10.1099/00222615-1-1-45. [DOI] [PubMed] [Google Scholar]
  39. Uhaa I. J., Hird D. W., Hirsh D. C., Jang S. S. Case-control study of risk factors associated with nosocomial Salmonella krefeld infection in dogs. Am J Vet Res. 1988 Sep;49(9):1501–1505. [PubMed] [Google Scholar]
  40. Vadivelu J., Dunn D. T., Feachem R. G., Drasar B. S., Cox N. P., Harrison T. J., Lloyd B. J. Comparison of five assays for the heat-labile enterotoxin of Escherichia coli. J Med Microbiol. 1987 May;23(3):221–226. doi: 10.1099/00222615-23-3-221. [DOI] [PubMed] [Google Scholar]
  41. Wasteson Y., Olsvik O., Skancke E., Bopp C. A., Fossum K. Heat-stable-enterotoxin-producing Escherichia coli strains isolated from dogs. J Clin Microbiol. 1988 Dec;26(12):2564–2566. doi: 10.1128/jcm.26.12.2564-2566.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weeratna R. D., Doyle M. P. Detection and production of verotoxin 1 of Escherichia coli O157:H7 in food. Appl Environ Microbiol. 1991 Oct;57(10):2951–2955. doi: 10.1128/aem.57.10.2951-2955.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Whipp S. C. Assay for enterotoxigenic Escherichia coli heat-stable toxin b in rats and mice. Infect Immun. 1990 Apr;58(4):930–934. doi: 10.1128/iai.58.4.930-934.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wilson J. B., McEwen S. A., Clarke R. C., Leslie K. E., Waltner-Toews D., Gyles C. L. A case-control study of selected pathogens including verocytotoxigenic Escherichia coli in calf diarrhea on an Ontario veal farm. Can J Vet Res. 1992 Jul;56(3):184–188. [PMC free article] [PubMed] [Google Scholar]
  45. Woodward M. J., Carroll P. J., Wray C. Detection of entero- and verocyto-toxin genes in Escherichia coli from diarrhoeal disease in animals using the polymerase chain reaction. Vet Microbiol. 1992 Jun 1;31(2-3):251–261. doi: 10.1016/0378-1135(92)90083-6. [DOI] [PubMed] [Google Scholar]
  46. Woodward M. J., Kearsley R., Wray C., Roeder P. L. DNA probes for the detection of toxin genes in Escherichia coli isolated from diarrhoeal disease in cattle and pigs. Vet Microbiol. 1990 Apr;22(2-3):277–290. doi: 10.1016/0378-1135(90)90115-c. [DOI] [PubMed] [Google Scholar]
  47. Woodward M. J., Wray C. Nine DNA probes for detection of toxin and adhesin genes in Escherichia coli isolated from diarrhoeal disease in animals. Vet Microbiol. 1990 Oct;25(1):55–65. doi: 10.1016/0378-1135(90)90093-b. [DOI] [PubMed] [Google Scholar]

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