Vet Med - Czech, 2009, 54(3):111-117 | DOI: 10.17221/3050-VETMED

Intermediately virulent Rhodococcus equi isolates from pigs in Slovenia: discovery of new plasmid types and assessment of genetic diversity by pulsed-field gel electrophoresis

M. Pate1, M. Ocepek1, I. Zdovc1, C. Minato2, Y. Ohtsu2, M. Matsuoka2, Y. Honda2, L. Hashimoto2, Y. Sasaki2, T. Kakuda2, S. Takai2
1 Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia
2 School of Veterinary Medicine, Kitasato University, Towada, Aomori, Japan

The presence of large plasmids in 30 Rhodococcus equi strains isolated from pig lymph nodes with granulomatous changes was investigated. Plasmid DNAs were isolated and digested with the restriction endonucleases BamHI, EcoRI, EcoT22I and HindIII for detailed comparison and estimation of plasmid sizes. A total of nine isolates were identified as intermediately virulent (VapB-positive), harbouring large plasmids of type 5 (n = 5) and four new variants that we tentatively designated as type 19 (n = 1), 20 (n = 1), 21 (n = 1) and 24 (n = 1). All isolates were subjected to genotyping with pulsed-field gel electrophoresis (PFGE). High genetic diversity was observed: 21 distinct genotypes were detected; five were found in multiple isolates and the others were unique. Isolates of the same plasmid type exhibited different PFGE profiles and vice versa. In a few cases, multiple strains from certain farms were analysed, the majority of which exhibited diverse PFGE profiles. Our findings demonstrate the presence of a wide variety of R. equi strains even in small confined environments such as farms. This is the first molecular epidemiology study of intermediately virulent R. equi isolates from Slovenian pigs.

Keywords: genotyping; molecular epidemiology; restriction enzyme analysis; virulence-associated genes

Published: March 31, 2009  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Pate M, Ocepek M, Zdovc I, Minato C, Ohtsu Y, Matsuoka M, et al.. Intermediately virulent Rhodococcus equi isolates from pigs in Slovenia: discovery of new plasmid types and assessment of genetic diversity by pulsed-field gel electrophoresis. Vet Med - Czech. 2009;54(3):111-117. doi: 10.17221/3050-VETMED.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Barton M.D., Hughes K.L. (1980): Corynebacterium equi: a review. Veterinary Bulletin, 50, 65-80.
    2. Bell K.S., Philp J.C., Christofi N., Aw D.W. (1996): Identification of Rhodococcus equi using the polymerase chain reaction. Letters in Applied Microbiology, 23, 72-74. Go to original source... Go to PubMed...
    3. Birnboim H.C., Doly J. (1979): A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research, 7, 1513-1523. Go to original source... Go to PubMed...
    4. Dvorska L., Parmova I., Lavickova M., Bartl J., Vrbas V., Pavlik I. (1999): Isolation of Rhodococcus equi and atypical mycobacteria from lymph nodes of pigs and cattle in herds with the occurrence of tuberculoid gross changes in the Czech Republic over the period of 1996-1998. Veterinarni Medicina, 44, 321-330.
    5. Fukunaga N., Okoda T., Katsumi M., Takai S. (1999): Restriction cleavage patterns of plasmid DNA of intermediately virulent Rhodococcus equi isolates from the mandibular lymph nodes of pigs in Kagoshima, Aomori and Miyagi Prefectures and the environment of pig farms. Journal of the Japan Veterinary Medical Association, 52, 789-792. Go to original source...
    6. Jones M.R., Neale T.J., Say P.J., Horne J.G. (1989): Rhodococcus equi: an emerging opportunistic pathogen? Australian and New Zealand Journal of Medicine, 19, 103-107. Go to original source... Go to PubMed...
    7. Karlson A.G., Moses H.E., Feldman W.H. (1940): Corynebacterium equi (Magnusson, 1923) in the submaxillary lymph nodes of swine. Journal of Infectious Diseases, 67, 243-251. Go to original source...
    8. Katsumi M., Kodama N., Miki Y., Hiramune T., Kikuchi N., Yanagawa R., Nakazawa M. (1991): Typing of Rhodococcus equi isolated from submaxillary lymph nodes of pigs in Japan. Zentralblatt fur Veterinarmedizin B, 38, 299-302. Go to original source... Go to PubMed...
    9. Kedlaya I., Ing M.B., Wong S.S. (2001): Rhodococcus equi infections in immunocompetent hosts: case report and review. Clinical Infectious Diseases, 32, E39-E46. Go to original source... Go to PubMed...
    10. Komijn R.E., Wisselink H.J., Rijsman V.M., StockhofeZurwieden N., Bakker D., van Zijderveld F.G., Eger T., Wagenaar J.A., Putirulan F.F., Urlings B.A. (2007): Granulomatous lesions in lymph nodes of slaughter pigs bacteriologically negative for Mycobacterium avium subsp. avium and positive for Rhodococcus equi. Veterinary Microbiology, 120, 352-357. Go to original source... Go to PubMed...
    11. Madarame H., Yaegashi R., Fukunaga N., Matsukuma M., Mutoh K., Morisawa N., Sasaki Y., Tsubaki S., Hasegawa Y., Takai S. (1998): Pathogenicity of Rhodococcus equi strains possessing virulence-associated 15- to 17-kDa and 20-kDa antigens: experimental and natural cases in pigs. Journal of Comparative Pathology, 119, 397-405. Go to original source... Go to PubMed...
    12. Makrai L., Takai S., Tamura M., Tsukamoto A., Sekimoto R., Sasaki Y., Kakuda T., Tsubaki S., Varga J., Fodor L., Solymosi N., Major A. (2002): Characterization of virulence plasmid types in Rhodococcus equi isolates from foals, pigs, humans and soil in Hungary. Veterinary Microbiology, 88, 377-384. Go to original source... Go to PubMed...
    13. Makrai L., Takayama S., Denes B., Hajtos I., Sasaki Y., Kakuda T., Tsubaki S., Major A., Fodor L., Varga J., Takai S. (2005): Characterization of virulence plasmids and serotyping of Rhodococcus equi isolates from submaxillary lymph nodes of pigs in Hungary. Journal of Clinical Microbiology, 43, 1246-1250. Go to original source... Go to PubMed...
    14. Makrai L., Kobayashi A., Matsouka M., Sasaki Y., Kakuda T., Denes B., Hajtos I., Revesz I., Janosi K., Fodor L., Varga J., Takai S. (2008): Isolation and characterization of Rhodococcus equi from submaxillary lymph nodes of wild boars (Sus scrofa). Veterinary Microbiology, doi:10.1016/j.vetmic.2008.04.009. Go to original source... Go to PubMed...
    15. Pate M., Zdovc I., Pirs T., Krt B., Ocepek M. (2004): Isolation and characterisation of Mycobacterium avium and Rhodococcus equi from granulomatous lesions of swine lymph nodes in Slovenia. Acta Veterinaria Hungarica, 52, 143-150. Go to original source... Go to PubMed...
    16. Prescott J.F. (1991): Rhodococcus equi: an animal and human pathogen. Clinical Microbiology Reviews, 4, 20-34. Go to original source... Go to PubMed...
    17. Ribeiro M.G., Seki I., Yasuoka K., Kakuda T., Sasaki Y., Tsubaki S., Takai S. (2005): Molecular epidemiology of virulent Rhodococcus equi from foals in Brazil: virulence plasmids of 85-kb type I, 87-kb type I, and a new variant, 87-kb type III. Comparative Immunology, Microbiology and Infectious Diseases, 28, 53-61. Go to original source...
    18. Shitaye J.E., Parmova I., Matlova L., Dvorska L., Horvathova A., Vrbas V., Pavlik I. (2006): Mycobacterial and Rhodococcus equi infections in pigs in the Czech Republic between the years 1996 and 2004: the causal factors and distribution of infections in the tissues. Veterinarni Medicina, 51, 497-511. Go to original source...
    19. Takai S. (1997): Epidemiology of Rhodococcus equi infections: a review. Veterinary Microbiology, 56, 167-176. Go to original source... Go to PubMed...
    20. Takai S., Tsubaki S. (1985): The incidence of Rhodococcus (Corynebacterium) equi in domestic animals and soil. Japanese Journal of Veterinary Science, 47, 493-496. Go to original source... Go to PubMed...
    21. Takai S., Watanabe Y., Ikeda T., Ozawa T., Matsukura S., Tamada Y., Tsubaki S., Sekizaki T. (1993): Virulenceassociated plasmids in Rhodococcus equi. Journal of Clinical Microbiology, 31, 1726-1729. Go to original source... Go to PubMed...
    22. Takai S., Imai Y., Fukunaga N., Uchida Y., Kamisawa K., Sasaki Y., Tsubaki S., Sekizaki T. (1995): Identification of virulence-associated antigens and plasmids in Rhodococcus equi from patients with AIDS. Journal of Infectious Diseases, 172, 1306-1311. Go to original source... Go to PubMed...
    23. Takai S., Fukunaga N., Ochiai S., Imai Y., Sasaki Y., Tsubaki S., Sekizaki T. (1996): Identification of intermediately virulent Rhodococcus equi isolates from pigs. Journal of Clinical Microbiology, 34, 1034-1037. Go to original source... Go to PubMed...
    24. Takai S., Shoda M., Sasaki Y., Tsubaki S., Fortier G., Pronost S., Rahal K., Becu T., Begg A., Browning G., Nicholson V.M., Prescott J.F. (1999): Restriction fragment length polymorphisms of virulence plasmids in Rhodococcus equi. Journal of Clinical Microbiology, 37, 3417-3420. Go to original source... Go to PubMed...
    25. Takai S., Tharavichitkul P., Sasaki C., Onishi Y., Yamano S., Kakuda T., Tsubaki S., Trinarong C., Rojanasthien S., Sirimalaisuwan A., Tesaprateep T., Maneekarn N., Sirisanthana T., Kirikae T. (2002): Identification of virulence-associated antigens and plasmids in Rhodococcus equi from patients with acquired immune deficiency syndrome and prevalence of virulent R. equi in soil collected from domestic animal farms in Chiang Mai, Thailand. American Journal of Tropical Medicine and Hygiene, 66, 52-55. Go to original source... Go to PubMed...
    26. Torres-Tortosa M., Arrizabalaga J., Villanueva J.L., Galvez J., Leyes M., Valencia M.E., Flores J., Pena J.M., Perez-Cecilia E., Quereda C. (2003): Prognosis and clinical evaluation of infection caused by Rhodococcus equi in HIV-infected patients: a multicenter study of 67 cases. Chest, 123, 1970-1976. Go to original source... Go to PubMed...
    27. Ulivieri S., Oliveri G. (2006): Cerebellar abscess due to Rhodococcus equi in an immunocompetent patient: case report and literature review. Journal of Neurosurgical Sciences, 50, 127-129. Go to PubMed...
    28. Weinstock D.M., Brown A.E. (2002): Rhodococcus equi: an emerging pathogen. Clinical Infectious Diseases, 34, 1379-1385. Go to original source... Go to PubMed...

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content