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BMC veterinary research2017; 13(1); 35; doi: 10.1186/s12917-017-0954-2

Molecular characterization of Rhodococcus equi isolates from horses in Poland: pVapA characteristics and plasmid new variant, 85-kb type V.

Abstract: Rhodococcus equi is one of the most significant bacterial pathogens affecting foals up to 6 months of age worldwide. Rhodococcosis is present in Poland however information about molecular characterization of R. equi isolates is scarce. This study describes molecular characterization of Rhodococcus equi infection on 13 horse breeding farms in Poland between 2001 and 2012. Samples were collected by tracheobronchial aspiration from pneumonic foals or during necropsy. The R. equi isolates were genotyped by plasmid profiling and pulsed-field gel electrophoresis. Results: Totally, 58 R. equi isolates were investigated. One isolate lost its plasmid. Among the 57 VapA-positive isolates, 48 contained 85-kb type I plasmid (82.8%), 8 contained 87-kb type I plasmid (13.8%). One isolate (1.7%) had a unique restriction cleavage pattern and the 2nd fragment of EcoRI digests of this plasmid DNA was about 2600 bases smaller than that of the 85 kb type I. This new plasmid variant was designated as the "85-kb type V". Among the 58 isolates typeable with VspI-PFGE, ten PFGE clusters were detected. The majority of foals were infected mostly with isolates of low genetic diversity. Conclusions: Most of clinical isolates of R. equi from foals in Poland contain pVapA 85-kb type I and 87-kb type I similarly to the other European countries and the United States. However, the new variant of pVapA 85-kb type V was identified. The chromosomal variability was detected among some of the investigated isolates and the presence of farm-specific isolates might be possible.
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Publication Date: 2017-01-26 PubMed ID: 28122544PubMed Central: PMC5267371DOI: 10.1186/s12917-017-0954-2Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study is about the molecular characterization of Rhodococcus equi – a bacteria causing infections in foals, from Poland. It provides valuable insights on the genetic diversity amongst these strains and identifies a new variant of the bacteria.

Overview of the Study

  • The study aims to thoroughly investigate and understand the molecular characteristics of Rhodococcus equi, a bacterial pathogen causing infections in young foals, isolated from Polish horse breeding farms.
  • The research was carried out across 13 horse breeding farms in Poland, between the years 2001 and 2012.
  • The bacteria strains were collected either from foals showing symptoms of pneumonia through tracheobronchial aspiration, or during post-mortem examinations.
  • These strains were then analyzed using plasmid profiling and pulsed-field gel electrophoresis, both are powerful genotyping techniques used for genetic mapping and studying the DNA of microbes.

Key Findings

  • A total of 58 R. equi strains were examined in the study. Among them, a plasmid was lost in one of the isolates.
  • Among the remaining 57 VapA-positive samples, 48 contained the 85-kb type I plasmid, representing about 82.8% of the strains, 8 samples contained the 87-kb type I plasmid, which is about 13.8% of the strains.
  • Interestingly, one of the isolates had a unique restriction cleavage pattern and was found to be around 2600 bases smaller than the 85 kb type I. This new, unique plasmid was named as the “85-kb type V”.
  • A total of ten different PFGE clusters were identified among the 58 isolates that were typed using VspI-PFGE.
  • The study also showed that the majority of foals were infected with isolates showing low genetic diversity and that there might be farm-specific isolates.

Conclusions

  • Like most European countries and the United States, the clinical isolates of R. equi from foals in Poland mostly contain pVapA 85-kb type I and 87-kb type I. However, the presence of a new variant, the pVapA 85-kb type V was identified.
  • The study also observed chromosomal variability across some of the investigated strains and suggested the possibility of the existence of farm-specific isolates.

Cite This Article

APA
Witkowski L, Rzewuska M, Takai S, Chrobak-Chmiel D, Kizerwetter-Świda M, Feret M, Gawryś M, Witkowski M, Kita J. (2017). Molecular characterization of Rhodococcus equi isolates from horses in Poland: pVapA characteristics and plasmid new variant, 85-kb type V. BMC Vet Res, 13(1), 35. https://doi.org/10.1186/s12917-017-0954-2

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 13
Issue: 1
Pages: 35
PII: 35

Researcher Affiliations

Witkowski, Lucjan
  • Laboratory of Veterinary Epidemiology and Economics, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159c, 02-776, Warsaw, Poland. lucjan_witkowski@sggw.pl.
Rzewuska, Magdalena
  • Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Ciszewskiego 8, 02-786, Warsaw, Poland.
Takai, Shinji
  • Department of Animal Hygiene, School of Veterinary Medicine, Kitasato University, Higashi 23-35-1, Towada, Aomori, 034-8628, Japan.
Chrobak-Chmiel, Dorota
  • Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Ciszewskiego 8, 02-786, Warsaw, Poland.
Kizerwetter-Świda, Magdalena
  • Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Ciszewskiego 8, 02-786, Warsaw, Poland.
Feret, Małgorzata
  • Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Ciszewskiego 8, 02-786, Warsaw, Poland.
Gawryś, Marta
  • Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Ciszewskiego 8, 02-786, Warsaw, Poland.
Witkowski, Maciej
  • University Center of Veterinary Medicine UJ-UR, Mickiewicza 24/28, 30-059, Cracow, Poland.
Kita, Jerzy
  • Laboratory of Veterinary Epidemiology and Economics, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159c, 02-776, Warsaw, Poland.

MeSH Terms

  • Actinomycetales Infections / epidemiology
  • Actinomycetales Infections / microbiology
  • Actinomycetales Infections / veterinary
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Gene Expression Regulation, Bacterial
  • Genetic Variation
  • Horse Diseases / epidemiology
  • Horse Diseases / microbiology
  • Horses
  • Poland
  • Rhodococcus equi / genetics
  • Rhodococcus equi / metabolism

References

This article includes 38 references
  1. Chaffin MK, Cohen ND, Martens RJ, Edwards RF, Nevill M. Foal-related risk factors associated with development of Rhodococcus equi pneumonia on farms with endemic infection.. J Am Vet Med Assoc 2003 Dec 15;223(12):1791-9.
    doi: 10.2460/javma.2003.223.1791pubmed: 14690209google scholar: lookup
  2. Muscatello G, Anderson GA, Gilkerson JR, Browning GF. Associations between the ecology of virulent Rhodococcus equi and the epidemiology of R. equi pneumonia on Australian thoroughbred farms.. Appl Environ Microbiol 2006 Sep;72(9):6152-60.
    doi: 10.1128/AEM.00495-06pmc: PMC1563629pubmed: 16957241google scholar: lookup
  3. Giguère S, Cohen ND, Chaffin MK, Hines SA, Hondalus MK, Prescott JF, Slovis NM. Rhodococcus equi: clinical manifestations, virulence, and immunity.. J Vet Intern Med 2011 Nov-Dec;25(6):1221-30.
    doi: 10.1111/j.1939-1676.2011.00804.xpubmed: 22092609google scholar: lookup
  4. Muscatello G. Rhodococcus equi pneumonia in the foal--part 1: pathogenesis and epidemiology.. Vet J 2012 Apr;192(1):20-6.
    doi: 10.1016/j.tvjl.2011.08.014pubmed: 22015138google scholar: lookup
  5. Cohen ND. Rhodococcus equi foal pneumonia.. Vet Clin North Am Equine Pract 2014 Dec;30(3):609-22.
    doi: 10.1016/j.cveq.2014.08.010pubmed: 25282322google scholar: lookup
  6. Yamshchikov AV, Schuetz A, Lyon GM. Rhodococcus equi infection.. Lancet Infect Dis 2010 May;10(5):350-9.
    doi: 10.1016/S1473-3099(10)70068-2pubmed: 20417417google scholar: lookup
  7. Witkowski L, Kaba J, Rzewuska M, Nowicki M, Szaluś-Jordanow O, Kita J. Development of ELISA test for determination of the level of antibodies against Rhodococcus equi in equine serum and colostrum.. Vet Immunol Immunopathol 2012 Oct 15;149(3-4):280-5.
    doi: 10.1016/j.vetimm.2012.07.009pubmed: 22883938google scholar: lookup
  8. Gradzki Z, Zietek-Barszcz A. Detection of Rhodococcus equi in tracheobronchial aspirate of foals in enzootic farms depending on age and season.. Bull Vet Inst Pulawy 2011;55(4):619–24.
  9. Banbura MW, Witkowski L, Chmielewska A, Tucholska A, Rzewuska M, Ruszczyk A. Isolation of equine herpes viruses type 1 and 2 (EHV-1 and EHV-2) from foals infected with Rhodocaccus equi.. Med Weter 2004;60(12):1333–6.
  10. Witkowski L, Rzewuska M, Takai S, Kizerwetter-Świda M, Kita J. Molecular epidemiology of Rhodococcus equi in slaughtered swine, cattle and horses in Poland.. BMC Microbiol 2016 May 27;16:98.
    doi: 10.1186/s12866-016-0712-9pmc: PMC4882809pubmed: 27234339google scholar: lookup
  11. Vázquez-Boland JA, Giguère S, Hapeshi A, MacArthur I, Anastasi E, Valero-Rello A. Rhodococcus equi: the many facets of a pathogenic actinomycete.. Vet Microbiol 2013 Nov 29;167(1-2):9-33.
    doi: 10.1016/j.vetmic.2013.06.016pubmed: 23993705google scholar: lookup
  12. Ocampo-Sosa AA, Lewis DA, Navas J, Quigley F, Callejo R, Scortti M, Leadon DP, Fogarty U, Vazquez-Boland JA. Molecular epidemiology of Rhodococcus equi based on traA, vapA, and vapB virulence plasmid markers.. J Infect Dis 2007 Sep 1;196(5):763-9.
    doi: 10.1086/519688pubmed: 17674320google scholar: lookup
  13. Venner M, Meyer-Hamme B, Verspohl J, Hatori F, Shimizu N, Sasaki Y, Kakuda T, Tsubaki S, Takai S. Genotypic characterization of VapA positive Rhodococcus equi in foals with pulmonary affection and their soil environment on a warmblood horse breeding farm in Germany.. Res Vet Sci 2007 Dec;83(3):311-7.
    doi: 10.1016/j.rvsc.2007.01.009pubmed: 17360011google scholar: lookup
  14. Duquesne F, Hébert L, Sévin C, Breuil MF, Tapprest J, Laugier C, Petry S. Analysis of plasmid diversity in 96 Rhodococcus equi strains isolated in Normandy (France) and sequencing of the 87-kb type I virulence plasmid.. FEMS Microbiol Lett 2010 Oct;311(1):76-81.
    doi: 10.1111/j.1574-6968.2010.02070.xpubmed: 20707817google scholar: lookup
  15. Makrai L, Takai S, Tamura M, Tsukamoto A, Sekimoto R, Sasaki Y, Kakuda T, Tsubaki S, Varga J, Fodor L, Solymosi N, Major A. Characterization of virulence plasmid types in Rhodococcus equi isolates from foals, pigs, humans and soil in Hungary.. Vet Microbiol 2002 Sep 24;88(4):377-84.
    doi: 10.1016/S0378-1135(02)00157-8pubmed: 12220812google scholar: lookup
  16. Takai S, Chaffin MK, Cohen ND, Hara M, Nakamura M, Kakuda T, Sasaki Y, Tsubaki S, Martens RJ. Prevalence of virulent Rhodococcus equi in soil from five R. equi-endemic horse-breeding farms and restriction fragment length polymorphisms of virulence plasmids in isolates from soil and infected foals in Texas.. J Vet Diagn Invest 2001 Nov;13(6):489-94.
    doi: 10.1177/104063870101300606pubmed: 11724139google scholar: lookup
  17. Takai S, Murata N, Kudo R, Narematsu N, Kakuda T, Sasaki Y, Tsubaki S. Two new variants of the Rhodococcus equi virulence plasmid, 90 kb type III and type IV, recovered from a foal in Japan.. Vet Microbiol 2001 Oct 1;82(4):373-81.
    doi: 10.1016/S0378-1135(01)00398-4pubmed: 11506930google scholar: lookup
  18. Takai S, Henton MM, Picard JA, Guthrie AJ, Fukushi H, Sugimoto C. Prevalence of virulent Rhodococcus equi in isolates from soil collected from two horse farms in South Africa and restriction fragment length polymorphisms of virulence plasmids in the isolates from infected foals, a dog and a monkey.. Onderstepoort J Vet Res 2001 Jun;68(2):105-10.
    pubmed: 11585087
  19. Ribeiro MG, Seki I, Yasuoka K, Kakuda T, Sasaki Y, Tsubaki S, Takai S. 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.. Comp Immunol Microbiol Infect Dis 2005 Jan;28(1):53-61.
    doi: 10.1016/S0147-9571(04)00045-1pubmed: 15563953google scholar: lookup
  20. Makrai L, Kira K, Kono A, Sasaki Y, Kakuda T, Tsubaki S, Fodor L, Varga J, Taka S. Plasmid profiles of virulent Rhodococcus equi isolates from soil environment on horse-breeding farms in Hungary.. Acta Vet Hung 2006 Mar;54(1):11-8.
    doi: 10.1556/AVet.54.2006.1.2pubmed: 16613022google scholar: lookup
  21. Witkowski L, Rzewuska M, Cisek AA, Chrobak-Chmiel D, Kizerwetter-Świda M, Czopowicz M, Welz M, Kita J. Prevalence and genetic diversity of Rhodococcus equi in wild boars (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in Poland.. BMC Microbiol 2015 May 22;15:110.
    doi: 10.1186/s12866-015-0445-1pmc: PMC4440313pubmed: 25997952google scholar: lookup
  22. Yuyama T, Yusa S, Yoshizumi K, Yamano S, Murata S, Hirose T, Osanai R, Onishi Y, Osato S, Sasaki C, Sasaki Y, Kakuda T, Tsubaki S, Takai S. Molecular epidemiology of VapA-positive Rhodococcus equi in thoroughbred horses in Kagoshima, Japan.. J Vet Med Sci 2002 Aug;64(8):715-8.
    doi: 10.1292/jvms.64.715pubmed: 12237518google scholar: lookup
  23. Son WG, Lee DS, Yamatoda N, Hatori F, Shimizu N, Kakuda T, Sasaki Y, Tsubaki S, Takai S. Molecular typing of VapA-positive Rhodococcus equi isolates from Jeju native horses, Korea.. J Vet Med Sci 2006 Mar;68(3):249-53.
    doi: 10.1292/jvms.68.249pubmed: 16598168google scholar: lookup
  24. Takai S, Son WG, Lee DS, Madarame H, Seki I, Yamatoda N, Kimura A, Kakuda T, Sasaki Y, Tsubaki S, Lim YK. Rhodococcus equi virulence plasmids recovered from horses and their environment in Jeju, Korea: 90-kb type II and a new variant, 90-kb type V.. J Vet Med Sci 2003 Dec;65(12):1313-7.
    doi: 10.1292/jvms.65.1313pubmed: 14709819google scholar: lookup
  25. Morton AC, Begg AP, Anderson GA, Takai S, Lämmler C, Browning GF. Epidemiology of Rhodococcus equi strains on Thoroughbred horse farms.. Appl Environ Microbiol 2001 May;67(5):2167-75.
    doi: 10.1128/AEM.67.5.2167-2175.2001pmc: PMC92851pubmed: 11319096google scholar: lookup
  26. Cohen ND, Smith KE, Ficht TA, Takai S, Libal MC, West BR, DelRosario LS, Becu T, Leadon DP, Buckley T, Chaffin MK, Martens RJ. Epidemiologic study of results of pulsed-field gel electrophoresis of isolates of Rhodococcus equi obtained from horses and horse farms.. Am J Vet Res 2003 Feb;64(2):153-61.
    doi: 10.2460/ajvr.2003.64.153pubmed: 12602583google scholar: lookup
  27. Bolton T, Kuskie K, Halbert N, Chaffin K, Healy M, Lawhon S, Jackson A, Cohen N. Detection of strain variation in isolates of Rhodococcus equi from an affected foal using repetitive sequence-based polymerase chain reaction.. J Vet Diagn Invest 2010 Jul;22(4):611-5.
    doi: 10.1177/104063871002200419pubmed: 20622235google scholar: lookup
  28. Takai S, Shoda M, Sasaki Y, Tsubaki S, Fortier G, Pronost S, Rahal K, Becu T, Begg A, Browning G, Nicholson VM, Prescott JF. Restriction fragment length polymorphisms of virulence plasmids in Rhodococcus equi.. J Clin Microbiol 1999 Oct;37(10):3417-20.
    pmc: PMC85590pubmed: 10488224doi: 10.1128/jcm.37.10.3417-3420.1999google scholar: lookup
  29. Takai S, Ohbushi S, Koike K, Tsubaki S, Oishi H, Kamada M. Prevalence of virulent Rhodococcus equi in isolates from soil and feces of horses from horse-breeding farms with and without endemic infections.. J Clin Microbiol 1991 Dec;29(12):2887-9.
    pmc: PMC270455pubmed: 1757567doi: 10.1128/jcm.29.12.2887-2889.1991google scholar: lookup
  30. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing.. J Clin Microbiol 1995 Sep;33(9):2233-9.
    pmc: PMC228385pubmed: 7494007doi: 10.1128/jcm.33.9.2233-2239.1995google scholar: lookup
  31. Pate M, Ocepek M, Zdovc I, Minato C, Ohtsu Y, Matsuoka M, Honda Y, Hashimoto L, Sasaki Y, Kakuda T. 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 (Praha) 2009;54(3):111–7.
  32. Soedarmanto I, Zhicai W, Setyamahanani A, Lämmler C. Pheno- and genotyping of Rhodococcus equi isolated from faeces of healthy horses and cattle.. Res Vet Sci 1998 May-Jun;64(3):181-5.
    doi: 10.1016/S0034-5288(98)90121-7pubmed: 9690599google scholar: lookup
  33. Morton AC, Baseggio N, Peters MA, Browning GF. Diversity of isolates of Rhodococcus equi from Australian thoroughbred horse farms.. Antonie Van Leeuwenhoek 1998 Jul-Oct;74(1-3):21-5.
    doi: 10.1023/A:1001791509073pubmed: 10068785google scholar: lookup
  34. Martens RJ, Takai S, Cohen ND, Chaffin MK, Liu H, Sakurai K, Sugimoto H, Lingsweiler SW. Association of disease with isolation and virulence of Rhodococcus equi from farm soil and foals with pneumonia.. J Am Vet Med Assoc 2000 Jul 15;217(2):220-5.
    doi: 10.2460/javma.2000.217.220pubmed: 10909463google scholar: lookup
  35. Rzewuska M, Witkowski L, Cisek AA, Stefańska I, Chrobak D, Stefaniuk E, Kizerwetter-Świda M, Takai S. Characterization of Rhodococcus equi isolates from submaxillary lymph nodes of wild boars (Sus scrofa), red deer (Cervus elaphus) and roe deer (Capreolus capreolus).. Vet Microbiol 2014 Aug 6;172(1-2):272-8.
    doi: 10.1016/j.vetmic.2014.04.020pubmed: 24878324google scholar: lookup
  36. Takai S, Watanabe Y, Ikeda T, Ozawa T, Matsukura S, Tamada Y, Tsubaki S, Sekizaki T. Virulence-associated plasmids in Rhodococcus equi.. J Clin Microbiol 1993 Jul;31(7):1726-9.
    pmc: PMC265621pubmed: 8349748doi: 10.1128/jcm.31.7.1726-1729.1993google scholar: lookup
  37. Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.. Nucleic Acids Res 1979 Nov 24;7(6):1513-23.
    doi: 10.1093/nar/7.6.1513pmc: PMC342324pubmed: 388356google scholar: lookup
  38. Takai S, Sekizaki T, Ozawa T, Sugawara T, Watanabe Y, Tsubaki S. Association between a large plasmid and 15- to 17-kilodalton antigens in virulent Rhodococcus equi.. Infect Immun 1991 Nov;59(11):4056-60.
    pmc: PMC258996pubmed: 1937765doi: 10.1128/iai.59.11.4056-4060.1991google scholar: lookup

Citations

This article has been cited 4 times.
  1. Żychska M, Witkowski L, Klementowska A, Rzewuska M, Kwiecień E, Stefańska I, Czopowicz M, Szaluś-Jordanow O, Mickiewicz M, Moroz A, Bonecka J, Kaba J. Rhodococcus equi-Occurrence in Goats and Clinical Case Report.. Pathogens 2021 Sep 4;10(9).
    doi: 10.3390/pathogens10091141pubmed: 34578172google scholar: lookup
  2. Rakowska A, Cywinska A, Witkowski L. Current Trends in Understanding and Managing Equine Rhodococcosis.. Animals (Basel) 2020 Oct 18;10(10).
    doi: 10.3390/ani10101910pubmed: 33081047google scholar: lookup
  3. Rzewuska M, Kwiecień E, Chrobak-Chmiel D, Kizerwetter-Świda M, Stefańska I, Gieryńska M. Pathogenicity and Virulence of Trueperella pyogenes: A Review.. Int J Mol Sci 2019 Jun 4;20(11).
    doi: 10.3390/ijms20112737pubmed: 31167367google scholar: lookup
  4. Kalinowski M, Grądzki Z, Jarosz Ł, Adaszek Ł. Molecular analysis of the chromosomal 16S rRNA gene and vapA plasmid gene of Polish field strains of R. equi.. PLoS One 2018;13(9):e0204024.
    doi: 10.1371/journal.pone.0204024pubmed: 30252885google scholar: lookup
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