1887

Abstract

The presence of the cytolethal distending toxin B gene () was examined in eight sp. flexispira reference strains, ATCC 700114 and 12 Finnish porcine strains and canine flexispira isolates. Part of the gene was amplified by PCR with degenerate primers VAT2 and DHF1, cloned and sequenced. The presence/absence of the gene as determined by PCR was confirmed by Southern hybridization and toxin production by HeLa cell-line experiments. PCR amplification resulted in approximately 700 bp fragments from sp. flexispira taxa 2 (ATCC 49314), 3 (ATCC 49320) and 8 (ATCC 43880, ATCC 49308, ATCC 43879), from six canine isolates as well as from the control strains and . The hybridization patterns of III-, dIII- and I-digested chromosomal DNA confirmed the results of the PCR experiments. The -positive strains had effects ranging from weak to strong on HeLa cell cultures. PCR amplification from the reference strains sp. flexispira taxa 1 (ATCC 43968), 4 (ATCC 49310) and 5 (ATCC 43966) and (ATCC 700114), and also six of the Finnish strains, was unsuccessful. No toxic effect on HeLa cells was evident when bacterial suspensions of PCR-negative strains were used for toxicity assay. Our results are in accordance with previous observations that the gene is not present in all species. Further, the presence/absence of the gene in sp. flexispira strains was in accordance with recent taxonomic analysis of the same strains, which suggests that it could serve as a useful marker in taxonomy.

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2003-02-01
2019-10-15
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References

  1. Bryner, J. H., Ritchie, A. E., Pollet, L., Kirkbride, C. A. & Collins, J. E. ( 1987;). Experimental infection and abortion of pregnant guinea pigs with a unique spirillum-like bacterium isolated from aborted ovine fetuses. Am J Vet Res 48, 91–95.
    [Google Scholar]
  2. Chien, C.-C., Taylor, N. S., Ge, Z., Schauer, D. B., Young, V. B. & Fox, J. G. ( 2000;). Identification of cdtB homologues and cytolethal distending toxin activity in enterohepatic Helicobacter spp. J Med Microbiol 49, 525–534.
    [Google Scholar]
  3. Comayras, C., Tasca, C., Pérès, S. Y., Ducommun, B., Oswald, E. & De Rycke, J. ( 1997;). Escherichia coli cytolethal distending toxin blocks the HeLa cell cycle at the G2/M transition by preventing cdc2 protein kinase dephosphorylation and activation. Infect Immun 65, 5088–5095.
    [Google Scholar]
  4. Dewhirst, F. E., Fox, J. G., Mendes, E. N., Paster, B. J., Gates, C. E., Kirkbride, C. A. & Eaton, K. A. ( 2000;). ‘Flexispira rappini’ strains represent at least 10 Helicobacter taxa. Int J Syst Evol Microbiol 50, 1781–1787.
    [Google Scholar]
  5. Felsenstein, J. ( 1985;). Confidence limits of phylogeny: an approach using the bootstrap. Evolution 39, 783–791.[CrossRef]
    [Google Scholar]
  6. Fox, J. G., Dewhirst, F. E., Tully, J. G., Paster, B. J., Yan, L., Taylor, N. S., Collins, M. J., Jr, Gorelick, P. L. & Ward, J. M. ( 1994;). Helicobacter hepaticus sp. nov., a microaerophilic bacterium isolated from livers and intestinal mucosal scrapings from mice. J Clin Microbiol 32, 1238–1245.
    [Google Scholar]
  7. Fox, J. G., Yan, L., Dewhirst, F. E., Paster, B. J., Shames, B., Murphy, J. C., Hayward, A., Belcher, J. C. & Mendes, E. N. ( 1995;). Helicobacter bilis sp. nov., a novel Helicobacter species isolated from bile, livers, and intestines of aged, inbred mice. J Clin Microbiol 33, 445–454.
    [Google Scholar]
  8. Fox, J. G., Chien, C. C., Dewhirst, F. E., Paster, B. J., Shen, Z., Melito, P. L., Woodward, D. L. & Rodgers, F. G. ( 2000;). Helicobacter canadensis sp. nov. isolated from humans with diarrhea as an example of an emerging pathogen. J Clin Microbiol 38, 2546–2549.
    [Google Scholar]
  9. Hänninen, M.-L., Happonen, I., Saari, S. & Jalava, K. ( 1996;). Culture and characteristics of Helicobacter bizzozeronii, a new canine gastric Helicobacter sp. Int J Syst Bacteriol 46, 160–166.[CrossRef]
    [Google Scholar]
  10. Hänninen, M.-L., Utriainen, M., Happonen, I. & Dewhirst, F. E. ( 2003;). Helicobacter sp. flexispira 16S rDNA taxa 1, 4 and 5 and Finnish porcine Helicobacter isolates are members of the species Helicobacter trogontum (taxon 6). Int J Syst Evol Microbiol 5 3 (in press).
    [Google Scholar]
  11. Jalava, K., On, S. L. W., Vandamme, P. A., Happonen, I., Sukura, A. & Hänninen, M. L. ( 1998;). Isolation and identification of Helicobacter spp. from canine and feline gastric mucosa. Appl Environ Microbiol 64, 3998–4006.
    [Google Scholar]
  12. Jalava, K., Hielm, S., Hirvi, U. & Hänninen, M.-L. (1999). Evaluation of a molecular identification scheme based on 23S rRNA gene polymorphisms for differentiating canine and feline gastric Helicobacter spp. Lett Appl Microbiol 28, 269–274.
  13. Kirkbride, C. A., Gates, C. E., Collins, J. E. & Ritchie, A. E. ( 1985;). Ovine abortion associated with an anaerobic bacterium. J Am Vet Med Assoc 186, 789–791.
    [Google Scholar]
  14. Kirkbride, C. A., Gates, C. E. & Collins, J. E. ( 1986;). Abortion in sheep caused by a nonclassified, anaerobic, flagellated bacterium. Am J Vet Res 47, 259–262.
    [Google Scholar]
  15. Lara-Tejero, M. & Galán, J. E. ( 2001;). CdtA, CdtB, and CdtC form a tripartite complex that is required for cytolethal distending toxin activity. Infect Immun 69, 4358–4365.[CrossRef]
    [Google Scholar]
  16. Mayer, M. P. A., Bueno, L. C., Hansen, E. J. & DiRienzo, J. M. ( 1999;). Identification of a cytolethal distending toxin gene locus and features of a virulence-associated region in Actinobacillus actinomycetemcomitans. Infect Immun 67, 1227–1237.
    [Google Scholar]
  17. Pickett, C. L., Pesci, E. C., Cottle, D. L., Russell, G., Erdem, A. N. & Zeytin, H. ( 1996;). Prevalence of cytolethal distending toxin production in Campylobacter jejuni and relatedness of Campylobacter sp. cdtB genes. Infect Immun 64, 2070–2078.
    [Google Scholar]
  18. Pitcher, D. G., Saunders, N. A. & Owen, R. J. ( 1989;). Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett Appl Microbiol 8, 151–156.[CrossRef]
    [Google Scholar]
  19. Romero, S., Archer, J. R., Hamacher, M. E., Bologna, S. M. & Schell, R. F. ( 1988;). Case report of an unclassified microaerophilic bacterium associated with gastroenteritis. J Clin Microbiol 26, 142–143.
    [Google Scholar]
  20. Swofford, D. L. (1996). paup: phylogenetic analysis using parsimony (and other methods), version 4.0. Sunderland, MA: Sinauer Associates.
  21. Taylor, N. S., Fox, J. G. & Yan, L. ( 1995;). In-vitro hepatotoxic factor in Helicobacter hepaticus, H. pylori and other Helicobacter species. J Med Microbiol 42, 48–52.[CrossRef]
    [Google Scholar]
  22. Young, V. B., Knox, K. A. & Schauer, D. B. ( 2000;a). Cytolethal distending toxin sequence and activity in the enterohepatic pathogen Helicobacter hepaticus. Infect Immun 68, 184–191.[CrossRef]
    [Google Scholar]
  23. Young, V. B., Chien, C.-C., Knox, K. A., Taylor, N. S., Schauer, D. B. & Fox, J. G. ( 2000;b). Cytolethal distending toxin in avian and human isolates of Helicobacter pullorum. J Infect Dis 182, 620–623.[CrossRef]
    [Google Scholar]
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