1887

Journal of Medical Microbiology logo

Volume 64, Issue 10

A new variant of cytolethal distending toxin in a clinical isolate of Free

Abstract

Increasing numbers of have been isolated from humans and animals with gastroenteritis, although the virulence mechanism of this species remains largely unknown. Here, we show that isolated from a patient with diarrhoea in Thailand produced a novel variant of cytolethal distending toxin (CDT). Sequencing of a 13 965 bp genomic region of carrying the genes coding for Ch-CDT revealed three ORFs of 798, 804 and 537 bp, which code for the Ch-CdtA, Ch-CdtB and Ch-CdtC subunits, respectively. The deduced amino acid sequence of Ch-CdtA showed ∼38.9 % homology with the CdtA of , but sequences of Ch-CdtB and Ch-CdtC were homologous to CdtB (65.7 %) and CdtC (33.1 %) of , respectively. Filter-sterilized sonic lysate of demonstrated distension and death of HeLa cells by arresting the cell cycle at the G/M phase and phosphorylation of host histone H2AX, a sensitive marker of DNA double-strand breaks. Rabbit antiserum raised against recombinant Ch-CdtB was not reactive against the recombinant CdtB protein of . A reconstituted Ch-CDT holotoxin prepared using each of the recombinant subunit proteins demonstrated distension and death of HeLa cells, suggesting that the isolate indeed produced functionally active Ch-CDT. Furthermore, the immunological distinctiveness of the Ch-CDT produced by and the increasing prevalence of the species in patients and animals with gastroenteritis suggest that this species may be an important emerging zoonotic pathogen.

  • Received:
  • Accepted:
  • Published Online:
© 2015 The Authors
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000145
2015-10-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/jmm/64/10/1124.html?itemId=/content/journal/jmm/10.1099/jmm.0.000145&mimeType=html&fmt=ahah

References

  1. Asakura M., Samosornsuk W., Taguchi M., Kobayashi K., Misawa N., Kusumoto M., Nishimura K., Matsuhisa A., Yamasaki S. 2007; Comparative analysis of cytolethal distending toxin (cdt) genes among Campylobacter jejuni, C. coli and C. fetus strains. Microb Pathog 42:174–183 [View Article][PubMed]
     [Google Scholar]
  2. Asakura M., Samosornsuk W., Hinenoya A., Misawa N., Nishimura K., Matsuhisa A., Yamasaki S. 2008; Development of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection and identification of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus . FEMS Immunol Med Microbiol 52:260–266 [View Article][PubMed]
     [Google Scholar]
  3. Bullman S., O'Leary J., Corcoran D., Sleator R. D., Lucey B. 2012; Molecular-based detection of non-culturable and emerging campylobacteria in patients presenting with gastroenteritis. Epidemiol Infect 140:684–688 [View Article][PubMed]
     [Google Scholar]
  4. Chaban B., Ngeleka M., Hill J. E. 2010; Detection and quantification of 14 Campylobacter species in pet dogs reveals an increase in species richness in feces of diarrheic animals. BMC Microbiol 10:73 [View Article][PubMed]
     [Google Scholar]
  5. Dasti J. I., Tareen A. M., Lugert R., Zautner A. E., Gross U. 2010; Campylobacter jejuni: a brief overview on pathogenicity-associated factors and disease-mediating mechanisms. Int J Med Microbiol 300:205–211 [View Article][PubMed]
     [Google Scholar]
  6. Edmonds P., Patton C. M., Griffin P. M., Barrett T. J., Schmid G. P., Baker C. N., Lambert M. A., Brenner D. J. 1987; Campylobacter hyointestinalis associated with human gastrointestinal disease in the United States. J Clin Microbiol 25:685–691[PubMed]
     [Google Scholar]
  7. Fennell C. L., Rompalo A. M., Totten P. A., Bruch K. L., Flores B. M., Stamm W. E. 1986; Isolation of “Campylobacter hyointestinalis” from a human. J Clin Microbiol 24:146–148[PubMed]
     [Google Scholar]
  8. Gebhart C. J., Ward G. E., Chang K., Kurtz H. J. 1983; Campylobacter hyointestinalis (new species) isolated from swine with lesions of proliferative ileitis. Am J Vet Res 44:361–367[PubMed]
     [Google Scholar]
  9. Gebhart C. J., Edmonds P., Ward G. E., Kurtz H. J., Brenner D. J. 1985; “Campylobacter hyointestinalis” sp. nov.: a new species of Campylobacter found in the intestines of pigs and other animals. J Clin Microbiol 21:715–720[PubMed]
     [Google Scholar]
  10. Gebhart C. J., Murtaugh M. P., Lin G. F., Ward G. E. 1990; Species-specific DNA probes for Campylobacter species isolated from pigs with proliferative enteritis. Vet Microbiol 24:367–379 [View Article][PubMed]
     [Google Scholar]
  11. Gharst G., Oyarzabal O. A., Hussain S. K. 2013; Review of current methodologies to isolate and identify Campylobacter spp. from foods. J Microbiol Methods 95:84–92 [View Article][PubMed]
     [Google Scholar]
  12. Gorkiewicz G., Feierl G., Zechner R., Zechner E. L. 2002; Transmission of Campylobacter hyointestinalis from a pig to a human. J Clin Microbiol 40:2601–2605 [View Article][PubMed]
     [Google Scholar]
  13. Hänninen M. L., Sarelli L., Sukura A., On S. L., Harrington C. S., Matero P., Hirvelä-Koski V. 2002; Campylobacter hyointestinalis subsp. hyointestinalis, a common Campylobacter species in reindeer. J Appl Microbiol 92:717–723 [View Article][PubMed]
     [Google Scholar]
  14. Hickey T. E., McVeigh A. L., Scott D. A., Michielutti R. E., Bixby A., Carroll S. A., Bourgeois A. L., Guerry P. 2000; Campylobacter jejuni cytolethal distending toxin mediates release of interleukin-8 from intestinal epithelial cells. Infect Immun 68:6535–6541 [View Article][PubMed]
     [Google Scholar]
  15. Hill B. D., Thomas R. J., Mackenzie A. R. 1987; Campylobacter hyointestinalis-associated enteritis in Moluccan rusa deer (Cervus timorensis subsp. Moluccensis). J Comp Pathol 97:687–694 [View Article][PubMed]
     [Google Scholar]
  16. Hinenoya A., Naigita A., Ninomiya K., Asakura M., Shima K., Seto K., Tsukamoto T., Ramamurthy T., Faruque S. M., Yamasaki S. 2009; Prevalence and characteristics of cytolethal distending toxin-producing Escherichia coli from children with diarrhea in Japan. Microbiol Immunol 53:206–215 [View Article][PubMed]
     [Google Scholar]
  17. Hinenoya A., Shima K., Asakura M., Nishimura K., Tsukamoto T., Ooka T., Hayashi T., Ramamurthy T., Faruque S. M., Yamasaki S. 2014; Molecular characterization of cytolethal distending toxin gene-positive Escherichia coli from healthy cattle and swine in Nara, Japan. BMC Microbiol 14:97 [View Article][PubMed]
     [Google Scholar]
  18. Inglis G. D., Boras V. F., Houde A. 2011; Enteric campylobacteria and RNA viruses associated with healthy and diarrheic humans in the Chinook health region of southwestern Alberta, Canada. J Clin Microbiol 49:209–219 [View Article][PubMed]
     [Google Scholar]
  19. Jain D., Prasad K. N., Sinha S., Husain N. 2008; Differences in virulence attributes between cytolethal distending toxin positive and negative Campylobacter jejuni strains. J Med Microbiol 57:267–272 [View Article][PubMed]
     [Google Scholar]
  20. Johnson W. M., Lior H. 1987; Response of Chinese hamster ovary cells to a cytolethal distending toxin (CDT) of Escherichia coli and possible misinterpretation as heat-labile (LT) enterotoxin. FEMS Microbiol Lett 43:19–23 [View Article]
     [Google Scholar]
  21. Kabir S. M., Kikuchi K., Asakura M., Shiramaru S., Tsuruoka N., Goto A., Hinenoya A., Yamasaki S. 2011; Evaluation of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the identification of Campylobacter strains isolated from diarrheal patients in Japan. Jpn J Infect Dis 64:19–27[PubMed]
     [Google Scholar]
  22. Lastovica A. J. 1996; Campylobacter/Helicobacter bacteraemia in Cape Town, South Africa, 1977–1995. In Campylobacters, Helicobacters and Related Organisms pp. 475–479 Edited by Newell D. G., Ketley J. M., Feldman R. A. New York: Plenum Press; [CrossRef]
     [Google Scholar]
  23. Lastovica A. J., Le Roux E. 2000; Efficient isolation of campylobacteria from stools. J Clin Microbiol 38:2798–2799[PubMed]
     [Google Scholar]
  24. Li L., Sharipo A., Chaves-Olarte E., Masucci M. G., Levitsky V., Thelestam M., Frisan T. 2002; The Haemophilus ducreyi cytolethal distending toxin activates sensors of DNA damage and repair complexes in proliferating and non-proliferating cells. Cell Microbiol 4:87–99 [View Article][PubMed]
     [Google Scholar]
  25. Man S. M. 2011; The clinical importance of emerging Campylobacter species. Nat Rev Gastroenterol Hepatol 8:669–685 [View Article][PubMed]
     [Google Scholar]
  26. Matsuda M., Shigematsu M., Tazumi A., Sekizuka T., Takamiya S., Millar B. C., Taneike I., Moore J. E. 2008; Cloning and structural analysis of the full-length cytolethal distending toxin (cdt) gene operon from Campylobacter lari . Br J Biomed Sci 65:195–199[PubMed]
     [Google Scholar]
  27. Minet J., Grosbois B., Megraud F. 1988; Campylobacter hyointestinalis: an opportunistic enteropathogen?. J Clin Microbiol 26:2659–2660[PubMed]
     [Google Scholar]
  28. Nakajima T., Tazumi A., Hirayama J., Hayashi K., Tasaki E., Asakura M., Yamasaki S., Moored J. E., Millar B. C., other authors. 2012; Expression and analysis of a cytolethal distending toxin (cdt) gene operon in Campylobacter lari . Br J Biomed Sci 6926–30
     [Google Scholar]
  29. Ohya T., Nakazawa M. 1992; Production and some properties of cytotoxins produced by Campylobacter species isolated from proliferative enteropathy in swine. J Vet Med Sci 54:1031–1033 [View Article][PubMed]
     [Google Scholar]
  30. Okuda J., Fukumoto M., Takeda Y., Nishibuchi M. 1997; Examination of diarrheagenicity of cytolethal distending toxin: suckling mouse response to the products of the cdtABC genes of Shigella dysenteriae . Infect Immun 65:428–433[PubMed]
     [Google Scholar]
  31. Oporto B., Hurtado A. 2011; Emerging thermotolerant Campylobacter species in healthy ruminants and swine. Foodborne Pathog Dis 8:807–813 [View Article][PubMed]
     [Google Scholar]
  32. 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 gene. Infect Immun 64:2070–2078[PubMed]
     [Google Scholar]
  33. Prouzet-Mauléon V., Labadi L., Bouges N., Ménard A., Mégraud F. 2006; Arcobacter butzleri: underestimated enteropathogen. Emerg Infect Dis 12:307–309 [View Article][PubMed]
     [Google Scholar]
  34. Purdy D., Buswell C. M., Hodgson A. E., McAlpine K., Henderson I., Leach S. A. 2000; Characterisation of cytolethal distending toxin (CDT) mutants of Campylobacter jejuni . J Med Microbiol 49:473–479[PubMed] [CrossRef]
     [Google Scholar]
  35. Russell R. G., Kiehlbauch J. A., Gebhart C. J., DeTolla L. J. 1992; Uncommon Campylobacter species in infant Macaca nemestrina monkeys housed in a nursery. J Clin Microbiol 30:3024–3027
     [Google Scholar]
  36. Samosornsuk W., Asakura M., Yoshida E., Taguchi T., Eampokalap B., Chaicumpa W., Yamasaki S. 2015; Isolation and characterization of Campylobacter strains from diarrheal patients in Bangkok and its suburb in Thailand. Jpn J Infect Dis 68:209–215 [View Article][PubMed]
     [Google Scholar]
  37. Shima A., Hinenoya A., Asakura M., Sugimoto N., Tsukamoto T., Ito H., Nagita A., Faruque S. M., Yamasaki S. 2012; Molecular characterizations of cytolethal distending toxin produced by Providencia alcalifaciens strains isolated from patients with diarrhea. Infect Immun 80:1323–1332 [View Article][PubMed]
     [Google Scholar]
  38. Vandenberg O., Dediste A., Houf K., Ibekwem S., Souayah H., Cadranel S., Douat N., Zissis G., Butzler J. P., Vandamme P. 2004; Arcobacter species in humans. Emerg Infect Dis 10:1863–1867 [View Article][PubMed]
     [Google Scholar]
  39. Vandenberg O., Houf K., Douat N., Vlaes L., Retore P., Butzler J. P., Dediste A. 2006; Antimicrobial susceptibility of clinical isolates of non-jejuni/coli campylobacters and arcobacters from Belgium. J Antimicrob Chemother 57:908–913 [View Article][PubMed]
     [Google Scholar]
  40. Yamasaki S., Asakura M., Tsukamoto T., Faruque S. M., Deb R., Ramamurthy T. 2006; Cytolethal distending toxin (CDT): genetic diversity, structure and role in diarrheal disease. Toxin Rev 25:61–88 [View Article]
     [Google Scholar]
  41. Yokoyama K. 2006; [Occurrence of Campylobacter food poisoning]. Jpn J Food Microbiol 23:109–113 (in Japanese)
    [Google Scholar]
  42. Young V. B., Knox K. A., Pratt J. S., Cortez J. S., Mansfield L. S., Rogers A. B., Fox J. G., Schauer D. B. 2004; In vitro and in vivo characterization of Helicobacter hepaticus cytolethal distending toxin mutants. Infect Immun 72:2521–2527 [View Article][PubMed]
     [Google Scholar]
  43. Yutsudo T., Nakabayashi N., Hirayama T., Takeda Y. 1987; Purification and some properties of a Vero toxin from Escherichia coli O157 : H7 that is immunologically unrelated to Shiga toxin. Microb Pathog 3:21–30 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000145
Loading
A new variant of cytolethal distending toxin in a clinical isolate of Campylobacter hyointestinalis
J. Med. Microbiol. 64, 1124 (2015); https://doi.org/10.1099/jmm.0.000145
/content/journal/jmm/10.1099/jmm.0.000145
/content/journal/jmm/10.1099/jmm.0.000145
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF

Most cited Most Cited RSS feed

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