1887

Abstract

The presence of bacteriophages encoding Shiga toxin 2 ( phages) was analysed in 168 strains of Shiga-toxin-producing (STEC) isolated from cattle. Following mitomycin C induction, strains carrying phages were screened by plaque blot and hybridization with an -probe. In the -phage-carrying strains, the amounts of phage production, phage DNA extracted and Stx produced after induction were assessed. The induced phages were characterized morphologically and genetically. Assays to obtain lysogens from different strains were also carried out and phages induced from the lysogens were compared with those induced from the STEC isolates. Results indicated that 18 % of the strains carried an inducible phage. Most of them showed a direct relationship between phage induction and toxin production. Each strain carried only one inducible phage, although a few strains had two copies of the in the chromosome. The phages showed diverse morphology and a wide variability in their genome. Assays to obtain lysogens showed that not all the phages were transduced with the same frequency and only six lysogens were obtained. Phages in the lysogens were the same as those induced from their respective initial STEC host strains, although the induction and relative toxin production of the lysogens varied. Most phages carried the gene, while a few carried variants. Infectivity of the phages depended on the different hosts, although O157 : H7 was preferentially infected by phages induced from O157 strains. The results show that inducible phages are common among STEC of animal origin and that they may enhance the spread of .

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27188-0
2004-09-01
2020-01-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/9/mic1502959.html?itemId=/content/journal/micro/10.1099/mic.0.27188-0&mimeType=html&fmt=ahah

References

  1. Acheson D. W. K., Reidl J., Zhang X., Keusch G. T., Mekalanos J. J., Waldor M. K. 1998; In vivo transduction with Shiga toxin 1-encoding phage. Infect Immun66:4496–4498
    [Google Scholar]
  2. Beutin L., Geier D., Zimmermann S., Aleksic S., Gillespie H. A., Whittam T. S. 1997; Epidemiological relatedness and clonal types of natural populations of Escherichia coli strains producing Shiga toxins in separate populations of cattle and sheep. Appl Environ Microbiol63:2175–2180
    [Google Scholar]
  3. Blanco J., Blanco M., Blanco J. E., Mora A., Alonso M. P., González E. A., Bernárdez M. I. 2001; Epidemiology of verocytotoxigenic Escherichia coli (VTEC) in ruminants. In Verocytotoxigenic Escherichia coli pp.113–148 Edited by Duffy G., Garvey P., McDowell D.. Trumbull, CT: Food & Nutrition Press;
    [Google Scholar]
  4. Blanco M., Blanco J. E., Mora A.. 9 other authors 2003; Serotypes, virulence genes and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolated from healthy sheep in Spain. J Clin Microbiol41:1351–1356[CrossRef]
    [Google Scholar]
  5. Brett K. N., Hornitzky M. A., Bettelheim K. A., Walker M. J., Djordjevic S. P. 2003; Bovine non-O157 Shiga toxin 2-containing Escherichia coli isolates commonly possess stx2-EDL933 and/or stx2vhb subtypes. J Clin Microbiol41:2716–2722[CrossRef]
    [Google Scholar]
  6. Corpet F. 1988; Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res16:10881–10890[CrossRef]
    [Google Scholar]
  7. Eklund M., Leino K., Siitonen A. 2002; Clinical Escherichia coli strains carrying stx genes: stx variants and stx-positive virulence profiles. J Clin Microbiol40:4585–4593[CrossRef]
    [Google Scholar]
  8. El-Sayed A. A. A. 2002; Molecular characterisation of two shigatoxin encoding bacteriophages from enterohaemorrhagic Escherichia coli Doctoral thesis Department of Veterinary Medicine, Justus-Liebig-University of Giessen; Germany:
    [Google Scholar]
  9. Franki R. I. B., Fauquet C. M., Knudson D. L., Brown F. 1991; Classification and nomenclature of viruses. 5th Report of theInternational Committee on Taxonomy of Viruses. Archives of Virology supplement 2 Vienna: Springer;
    [Google Scholar]
  10. Guinée P. A. M., Jansen W. H., Wadström T., Sellwood R. 1981; Escherichia coli associated with neonatal diarrhea in piglets and calves. Curr Top Vet Anim Sci13:126–162
    [Google Scholar]
  11. Hancock D., Besser T., Lejeune J., Davis M., Rice D. 2001; The control of VTEC in the animal reservoir. Int J Food Microbiol66:71–78[CrossRef]
    [Google Scholar]
  12. Heuvelink A. E., van den Biggeelaar F. L., Zwartkruis-Nahuis J., Herbes R. G., Huyben R., Nagelkerke N., Melchers W. J., Monnens L. A., de Boer E. 1998; Occurrence of verocytotoxin-producing Escherichia coli O157 on Dutch dairy farms. J Clin Microbiol36:3480–3487
    [Google Scholar]
  13. James C. E., Stanley K. N., Allison H. E., Flint H. J., Stewart C. S., Sharp R. J., Saunders J. R., McCarthy A. J. 2001; Lytic and lysogenic infection of diverse Escherichia coli and Shigella strains with a verocytotoxigenic bacteriophage. Appl Environ Microbiol67:4335–4337[CrossRef]
    [Google Scholar]
  14. Johansen B. K., Wasteson Y., Granum P. E., Brynestad S. 2001; Mosaic structure of Shiga-toxin-2 encoding phages isolated from Escherichia coli O157 : H7 indicates frequent gene exchange between lambdoid phage genomes. Microbiology147:1929–1936
    [Google Scholar]
  15. Karmali M. A. 2004; Infection by Shiga toxin-producing Escherichia coli: an overview. Mol Biotechnol26:117–122[CrossRef]
    [Google Scholar]
  16. Kimmitt P. T., Harwood C. R., Barer M. R. 2000; Toxin gene expression by Shiga toxin-producing Escherichia coli: the role of antibiotics and the bacterial SOS response. Emerg Infect Dis6:458–465[CrossRef]
    [Google Scholar]
  17. Koch C., Hertwig S., Lurz R., Appel B., Beutin L. 2001; Isolation of a lysogenic bacteriophage carrying the stx1OX3 gene, which is closely associated with Shiga toxin-producing Escherichia coli strains from sheep and humans. J Clin Microbiol39:3992–3998[CrossRef]
    [Google Scholar]
  18. Kohler B., Karch H., Schmidt H. 2000; Antibacterials that are used as growth promoters in animal husbandry can affect the production of Shiga-toxin-2-converting bacteriophages and Shiga toxin 2 from Escherichia coli strains. Microbiology146:1085–1090
    [Google Scholar]
  19. Licence K., Oates K. R., Synge B. A., Reid T. M. S. 2001; An outbreak of E. coli O157 infection with evidence of spread from animals to man through contamination of a private water supply. Epidemiol Infect126:135–138
    [Google Scholar]
  20. Livny J., Friedman D. I. 2004; Characterizing spontaneous induction of Stx encoding phages using a selectable reporter system. Mol Microbiol51:1691–1704[CrossRef]
    [Google Scholar]
  21. Meyer T. H., Karch H., Hacker J., Bocklage H., Heesemann J. 1992; Cloning and sequencing of a Shiga-like toxin II-related gene from Escherichia coli O157 : H7 strain 7279. Zentralbl Bakteriol276:176–188[CrossRef]
    [Google Scholar]
  22. Muniesa M., Jofre J. 1998; Abundance in sewage of bacteriophages that infect E. coli O157 : H7 and that carry the Shiga toxin 2 gene. Appl Environ Microbiol64:2443–2448
    [Google Scholar]
  23. Muniesa M., Jofre J. 2000; Occurrence of phages infecting Escherichia coli O157 : H7 carrying the Stx2 gene in sewage from different countries. FEMS Microbiol Lett183:197–200[CrossRef]
    [Google Scholar]
  24. Muniesa M., Recktenwald J., Bielaszewska M., Karch H., Schmidt H. 2000; Characterization of a Shiga toxin 2e-converting bacteriophage from an Escherichia coli strain of human origin. Infect Immun68:4850–4855[CrossRef]
    [Google Scholar]
  25. Muniesa M., Mocé-Llivina L., Katayama H., Jofre J. 2003; Bacterial host strains that support replication of somatic coliphages. Antonie Van Leeuwenhoek83:305–315[CrossRef]
    [Google Scholar]
  26. Muniesa M., Serra-Moreno R., Jofre J. 2004; Free Shiga toxin bacteriophages isolated from sewage showed diversity although the stx genes appeared conserved. Environ Microbiol6:716–725[CrossRef]
    [Google Scholar]
  27. Nataro J. P., Kaper J. B. 1998; Diarrheagenic Escherichia coli. Clin Microbiol Rev11:142–201
    [Google Scholar]
  28. Neely M. N., Friedman D. I. 1998; Functional and genetic analysis of regulatory regions of coliphage H-19B: location of Shiga-like toxin and lysis genes suggest a role for phage functions in toxin release. Mol Microbiol28:1255–1267[CrossRef]
    [Google Scholar]
  29. O'Brien A. D., Newland J. W., Miller S. F., Holmes R. K., Smith H. W., Formal S. B. 1984; Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea. Science226:694–696[CrossRef]
    [Google Scholar]
  30. Paton J. C., Paton A. W. 1998; Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli infections. Clin Microbiol Rev11:450–479
    [Google Scholar]
  31. Plunkett G., Rose D. J., Durfee T. J., Blattner F. R 3rd. 1999; Sequence of Shiga toxin 2 phage 933W from Escherichia coli O157 : H7: Shiga toxin as a phage late-gene product. J Bacteriol181:1767–1778
    [Google Scholar]
  32. Ritchie J. M., Wagner P. L., Acheson D. W. K., Waldor M. K. 2003; Comparison of Shiga toxin production by hemolytic-uremic syndrome-associated and bovine-associated Shiga toxin-producing Escherichia coli isolates. Appl Environ Microbiol69:1059–1066[CrossRef]
    [Google Scholar]
  33. Rüssmann, Schmidt H., Caprioli A., Karch H. 1994; Highly conserved B-subunit genes of Shiga-like toxin II variants found in Escherichia coli O157 strains. FEMS Microbiol Lett118:335–340[CrossRef]
    [Google Scholar]
  34. Sambrook J., Russell D. W. 2001; Molecular Cloning: a Laboratory Manual, 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  35. Schmidt H., Bielaszewska M., Karch H. 1999; Transduction of enteric Escherichia coli isolates with a derivative of Shiga toxin 2-encoding bacteriophage π3538 isolated fromE. coli O157 : H7. Appl Environ Microbiol65:3855–3861
    [Google Scholar]
  36. Schmidt H., Scheef J., Morabito S., Caprioli A., Wieler L. H., Karch H. 2000; A new Shiga toxin 2 variant (Stx2f) from Escherichia coli isolated from pigeons. Appl Environ Microbiol66:1205–1208[CrossRef]
    [Google Scholar]
  37. Stewart D. S., Tortorello M. L., Gendel S. M. 1998; Evaluation of DNA preparation techniques for detection of the SLT-1 gene of Escherichia coli O157 : H7 in bovine faeces using the polymerase chain reaction. Lett Appl Microbiol26:93–97[CrossRef]
    [Google Scholar]
  38. Strauch E., Lurz R., Beutin L. 2001; Characterization of a Shiga toxin-encoding temperate bacteriophage of Shigella sonnei. Infect Immun69:7588–7595[CrossRef]
    [Google Scholar]
  39. Tanji Y., Mizoguchi K., Yoichi M., Morita M., Kijima N., Kator H., Unno H. 2003; Seasonal change and fate of coliphages infected to Escherichia coli O157 : H7 in a wastewater treatment plant. Water Res37:1136–1142[CrossRef]
    [Google Scholar]
  40. Teel L. D., Melton-Celsa A. R., Schmitt C. K., O'Brien A. D. 2002; One of two copies of the gene for the activatable Shiga toxin type 2d in Escherichia coli O91 : H21 strain B2F1 is associated with an inducible bacteriophage. Infect Immun70:4282–4291[CrossRef]
    [Google Scholar]
  41. Unkmeir A., Schmidt H. 2000; Structural analysis of phage-borne stx genes and their flanking sequences in Shiga toxin-producingEscherichia coli and Shigella dysenteriae type 1 strains. Infect Immun68:4856–4864[CrossRef]
    [Google Scholar]
  42. Wagner P. L., Acheson D. W., Waldor M. K. 1999; Isogenic lysogens of diverse Shiga toxin 2-encoding bacteriophages produce markedly different amounts of Shiga toxin. Infect Immun67:6710–6714
    [Google Scholar]
  43. Widiasih D. A., Ido N., Omoe K., Sugii S., Shinagawa K. 2004; Duration and magnitude of faecal shedding of Shiga toxin-producing Escherichia coli from naturally infected cattle. Epidemiol Infect132:67–75[CrossRef]
    [Google Scholar]
  44. WHO 1991; Subcommittee meeting held at the conference ‘Shiga-like toxin producing Escherichia coli with a special emphasis on zoonotic aspects’ Giessen; Germany:
    [Google Scholar]
  45. WHO 2001; Monitoring antimicrobial usage in food animals for the protection of human health Report of a WHO consultation 10–13 September 2001; Available athttp://www.who.int/emc/diseases/zoo/antimicrobial.html
    [Google Scholar]
  46. Zhang X., McDaniel A. D., Wolf L. E., Keusch G. T., Waldor M. K., Acheson D. W. 2000; Quinolone antibiotics induce Shiga toxin-encoding bacteriophages, toxin production, and death in mice. J Infect Dis181:664–670[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.27188-0
Loading
/content/journal/micro/10.1099/mic.0.27188-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error