Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1991 Nov;29(11):2522–2527. doi: 10.1128/jcm.29.11.2522-2527.1991

Virulence factors associated with cytotoxic necrotizing factor type two in bovine diarrheic and septicemic strains of Escherichia coli.

E Oswald 1, J de Rycke 1, P Lintermans 1, K van Muylem 1, J Mainil 1, G Daube 1, P Pohl 1
PMCID: PMC270366  PMID: 1774259

Abstract

Forty-three bovine isolates of Escherichia coli producing a second type of cytotoxic necrotizing factor (CNF2) and three K-12 strains carrying different Vir plasmids coding for CNF2 were tested for the presence of several virulence factors. Most of the strains were serum resistant (79%), produced an aerobactin (70%), and adhered to calf villi (53%); some of them produced a colicin (32%) and a hemolysin (9%). These strains were also tested by a colony hybridization assay with gene probes for six toxins (classical heat-stable [STaP and STb] and heat-labile [LT-I and LT-IIa] enterotoxins and Shiga-like toxins [SLT-I and SLT-II]) and five adhesion factors (K99, K88, 987P, F17, and F41). Only two gene probes, LT-IIa (9%) and F17A (53%), hybridized with the CNF2 strains. However, antibodies raised against F17 fimbriae did not agglutinate the strains hybridizing with the F17A probe. In contrast, all except one of these strains adhered to calf villi. Interestingly, these two properties, F17A positivity and adherence to calf villi, were the only ones expressed by the K-12 strains carrying different Vir plasmids. In conclusion, this study confirmed that CNF2-producing strains are unrelated to previously described toxigenic E. coli strains and also demonstrated that in half of the strains the production of CNF2 was associated with an adhesion factor genetically related to, but different from, F17, which is more than likely encoded by Vir plasmids.

Full text

PDF
2522

Selected References

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

  1. Anderson D. G., Moseley S. L. Escherichia coli F41 adhesin: genetic organization, nucleotide sequence, and homology with the K88 determinant. J Bacteriol. 1988 Oct;170(10):4890–4896. doi: 10.1128/jb.170.10.4890-4896.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bisicchia R., Ciammarughi R., Caprioli A., Falbo V., Ruggeri F. M. Toxin production and haemagglutination in strains of Escherichia coli from diarrhoea in Brescia, Italy. J Hyg (Lond) 1985 Oct;95(2):353–361. doi: 10.1017/s002217240006277x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blanco J., González E. A., García 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 Dec;18(3-4):297–311. doi: 10.1016/0378-1135(88)90095-8. [DOI] [PubMed] [Google Scholar]
  4. Caprioli A., Falbo V., Roda L. G., Ruggeri F. M., Zona C. Partial purification and characterization of an escherichia coli toxic factor that induces morphological cell alterations. Infect Immun. 1983 Mar;39(3):1300–1306. doi: 10.1128/iai.39.3.1300-1306.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Contrepois M., Dubourguier H. C., Parodi A. L., Girardeau J. P., Ollier J. L. Septicaemic Escherichia coli and experimental infection of calves. Vet Microbiol. 1986 Jul;12(2):109–118. doi: 10.1016/0378-1135(86)90073-8. [DOI] [PubMed] [Google Scholar]
  6. Contrepois M., Martel J. L., Bordas C., Hayers F., Millet A., Ramisse J., Sendral R. Fréquence des pili FY et K99 parmi des souches de Escherichia coli isolées de veaux diarrhéiques en France. Ann Rech Vet. 1985;16(1):25–28. [PubMed] [Google Scholar]
  7. Cross M. A., Koronakis V., Stanley P. L., Hughes C. HlyB-dependent secretion of hemolysin by uropathogenic Escherichia coli requires conserved sequences flanking the chromosomal hly determinant. J Bacteriol. 1990 Mar;172(3):1217–1224. doi: 10.1128/jb.172.3.1217-1224.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dallas W. S., Gill D. M., Falkow S. Cistrons encoding Escherichia coli heat-labile toxin. J Bacteriol. 1979 Sep;139(3):850–858. doi: 10.1128/jb.139.3.850-858.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Rycke J., González E. A., Blanco J., Oswald E., Blanco M., Boivin R. Evidence for two types of cytotoxic necrotizing factor in human and animal clinical isolates of Escherichia coli. J Clin Microbiol. 1990 Apr;28(4):694–699. doi: 10.1128/jcm.28.4.694-699.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Rycke J., Guillot J. F., Boivin R. Cytotoxins in non-enterotoxigenic strains of Escherichia coli isolated from feces of diarrheic calves. Vet Microbiol. 1987 Oct;15(1-2):137–150. doi: 10.1016/0378-1135(87)90139-8. [DOI] [PubMed] [Google Scholar]
  11. De Rycke J., Oswald E., Boivin R. An in vivo assay for the detection of cytotoxic strains of Escherichia coli. Ann Rech Vet. 1989;20(1):39–46. [PubMed] [Google Scholar]
  12. FREDERICQ P. Colicins. Annu Rev Microbiol. 1957;11:7–22. doi: 10.1146/annurev.mi.11.100157.000255. [DOI] [PubMed] [Google Scholar]
  13. Fierer J., Finley F., Braude A. I. A plaque assay on agar for detection of gram-negative bacilli sensitive to complement. J Immunol. 1972 Nov;109(5):1156–1158. [PubMed] [Google Scholar]
  14. Girardeau J. P. A new in vitro technique for attachment to intestinal villi using enteropathogenic Escherichia coli. Ann Microbiol (Paris) 1980 Jul-Aug;131B(1):31–37. [PubMed] [Google Scholar]
  15. Guinée P. A., Veldkamp J., Jansen W. H. Improved minca medium for the detection of K99 antigen in calf enterotoxigenic strains of Escherichia coli. Infect Immun. 1977 Feb;15(2):676–678. doi: 10.1128/iai.15.2.676-678.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kehoe M., Sellwood R., Shipley P., Dougan G. Genetic analysis of K88-mediated adhesion of enterotoxigenic Escherichia coli. Nature. 1981 May 14;291(5811):122–126. doi: 10.1038/291122a0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Lintermans P. F., Pohl P., Bertels A., Charlier G., Vandekerckhove J., Van Damme J., Schoup J., Schlicker C., Korhonen T., De Greve H. Characterization and purification of the F17 adhesin on the surface of bovine enteropathogenic and septicemic Escherichia coli. Am J Vet Res. 1988 Nov;49(11):1794–1799. [PubMed] [Google Scholar]
  19. Lintermans P., Pohl P., Deboeck F., Bertels A., Schlicker C., Vandekerckhove J., Van Damme J., Van Montagu M., De Greve H. Isolation and nucleotide sequence of the F17-A gene encoding the structural protein of the F17 fimbriae in bovine enterotoxigenic Escherichia coli. Infect Immun. 1988 Jun;56(6):1475–1484. doi: 10.1128/iai.56.6.1475-1484.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lopez-Alvarez J., Gyles C. L. Occurrence of the vir plasmid among animal and human strains of invasive Escherichia coli. Am J Vet Res. 1980 May;41(5):769–774. [PubMed] [Google Scholar]
  21. Lopez-Alvarez J., Gyles C. L., Shipley P. L., Falkow S. Genetic and molecular characteristics of Vir plasmids of bovine septicemic Escherichia coli. J Bacteriol. 1980 Feb;141(2):758–769. doi: 10.1128/jb.141.2.758-769.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mainil J. G., Bex F., Jacquemin E., Pohl P., Couturier M., Kaeckenbeeck A. Prevalence of four enterotoxin (STaP, STaH, STb, and LT) and four adhesin subunit (K99, K88, 987P, and F41) genes among Escherichia coli isolates from cattle. Am J Vet Res. 1990 Feb;51(2):187–190. [PubMed] [Google Scholar]
  23. Mainil J. G., Moseley S. L., Schneider R. A., Sutch K., Casey T. A., Moon H. W. Hybridization of bovine Escherichia coli isolates with gene probes for four enterotoxins (STaP, STaH, STb, LT) and one adhesion factor (K99). Am J Vet Res. 1986 May;47(5):1145–1148. [PubMed] [Google Scholar]
  24. Mainil J., Bex F., Couturier M., Kaeckenbeeck A. Hybridization on bovine enterotoxigenic Escherichia coli with two heat-stable enterotoxin gene probes. Am J Vet Res. 1985 Dec;46(12):2582–2584. [PubMed] [Google Scholar]
  25. Morris J. A., Thorns C. J., Scott A. C., Sojka W. J. Adhesive properties associated with the Vir plasmid: a transmissible pathogenic characteristic associated with strains of invasive Escherichia coli. J Gen Microbiol. 1982 Sep;128(9):2097–2103. doi: 10.1099/00221287-128-9-2097. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Oswald E., De Rycke J. A single protein of 110 kDa is associated with the multinucleating and necrotizing activity coded by the Vir plasmid of Escherichia coli. FEMS Microbiol Lett. 1990 Mar 15;56(3):279–284. [PubMed] [Google Scholar]
  28. Oswald E., De Rycke J., Guillot J. F., Boivin R. Cytotoxic effect of multinucleation in HeLa cell cultures associated with the presence of Vir plasmid in Escherichia coli strains. FEMS Microbiol Lett. 1989 Mar;49(1):95–99. doi: 10.1016/0378-1097(89)90349-2. [DOI] [PubMed] [Google Scholar]
  29. Pickett C. L., Twiddy E. M., Belisle B. W., Holmes R. K. Cloning of genes that encode a new heat-labile enterotoxin of Escherichia coli. J Bacteriol. 1986 Feb;165(2):348–352. doi: 10.1128/jb.165.2.348-352.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pohl P., Lintermans P., Mainil J., Daube G., Kaeckenbeeck A. ETEC-like strains from cattle. Vet Rec. 1989 Sep 30;125(14):382–382. doi: 10.1136/vr.125.14.382. [DOI] [PubMed] [Google Scholar]
  31. Shimizu M., Sakano T., Yamamoto J., Kitajima K. Incidence and some characteristics of fimbriae FY and 31A of Escherichia coli isolates from calves with diarrhea in Japan. Microbiol Immunol. 1987;31(5):417–426. doi: 10.1111/j.1348-0421.1987.tb03104.x. [DOI] [PubMed] [Google Scholar]
  32. Smith H. W. A search for transmissible pathogenic characters in invasive strains of Escherichia coli: the discovery of a plasmid-controlled toxin and a plasmid-controlled lethal character closely associated, or identical, with colicine V. J Gen Microbiol. 1974 Jul;83(0):95–111. doi: 10.1099/00221287-83-1-95. [DOI] [PubMed] [Google Scholar]
  33. Valvano M. A., Crosa J. H. Aerobactin iron transport genes commonly encoded by certain ColV plasmids occur in the chromosome of a human invasive strain of Escherichia coli K1. Infect Immun. 1984 Oct;46(1):159–167. doi: 10.1128/iai.46.1.159-167.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Welch R. A., Dellinger E. P., Minshew B., Falkow S. Haemolysin contributes to virulence of extra-intestinal E. coli infections. Nature. 1981 Dec 17;294(5842):665–667. doi: 10.1038/294665a0. [DOI] [PubMed] [Google Scholar]
  35. de Graaf F. K., Klaasen P. Organization and expression of genes involved in the biosynthesis of 987P fimbriae. Mol Gen Genet. 1986 Jul;204(1):75–81. doi: 10.1007/BF00330190. [DOI] [PubMed] [Google Scholar]

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

RESOURCES