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Home / Equine Research Bank / Curr Microbiol / Molecular characterization of Rhodococcus equi from horse-br...
Current microbiology2009; 58(4); 399-403; doi: 10.1007/s00284-009-9370-6

Molecular characterization of Rhodococcus equi from horse-breeding farms by means of multiplex PCR for the vap gene family.

Abstract: This study evaluated the molecular characteristics of Rhodococcus equi isolates obtained from horses by a multiplex PCR assay that amplifies the vap gene family (vapA, -B, -C, -D, -E, -F, -G, and -H). A total of 180 R. equi isolates were studied from four different sources, namely healthy horse feces (112), soil (12), stalls (23), and clinical isolates (33) from horse-breeding farms. The technique was performed and confirmed by sequencing of amplified vap gene family controls. Thirty-two (17.8%) of the R. equi isolates were positive for the vapA gene and carried at least three other vap genes. All 147 isolates from equine feces, stalls, and soil failed to demonstrate any genes associated with virulence-inducing proteins. About 32 (97.0%) out of the 33 clinical equine isolates tested positive for the multiplex PCR assay for the vap gene family. They demonstrated six molecular profiles: 100% featured the vapA, vapD, and vapG genes, 86.6% vapF, 76.6% vapH, 43.3% vapC, 36.6% vapE, and none vapB. The most frequent molecular profile was vap A, -D, -F, G, and -H, where this profile was present in 37.5% of the strains. Moreover, there was no molecular epidemiological pattern for R. equi isolates that uniquely mapped to each horse-breeding farm studied. Our proposed technique allows the identification of eight members of the vap gene family (vapA, B, -C, -D, -E, -F, -G, and -H). It is a practical and efficient method of conducting clinical and epidemiological studies on R. equi isolates.
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Publication Date: 2009-02-10 PubMed ID: 19205798DOI: 10.1007/s00284-009-9370-6Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

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 research article discusses a study that used multiplex PCR to assess the genetic characteristics of Rhodococcus equi bacteria isolated from horses and their environment. By doing so, the researchers were able to identify different profiles of virulence-associated protein (vap) genes in different isolates, without finding a unique pattern specific to each breeding farm.

Overview of Research Methods and Findings

  • The researchers used a technique called multiplex PCR, which stands for polymerase chain reaction. This technique provides a way to amplify specific sections of DNA, in this case the vap gene family in Rhodococcus equi bacteria.
  • The researchers applied this technique on 180 isolates of R. equi obtained from four different sources: healthy horse feces, soil, stalls, and clinical isolates from sick horses from horse-breeding farms.
  • To ensure the accuracy and reliability of the PCR technique, the researchers also conducted sequencing on the amplified DNA samples as a form of control.

Specific Findings Regarding the vap Gene Family

  • About 18% of the R. equi isolates carried the vapA gene along with at least three other members of the vap gene family.
  • All isolates from healthy equine feces, stalls, and soil did not show any genes associated with virulence-inducing proteins.
  • Almost all (97%) of the clinical isolates from diseased horses tested positive for the vap gene family.
  • The researchers were able to identify six different molecular profiles among these clinical isolates based on the presence of different vap genes.

Implications of the Research Findings

  • The research findings represent significant progress in identifying genetic markers of R. equi bacteria, a common cause of pneumonia in foals.
  • However, the researchers did not find a unique molecular epidemiological pattern for R. equi from each horse-breeding farm, suggesting that strain differences are not strictly farm-specific.
  • The proposed method is efficient for identifying members of the vap gene family and may be used for future clinical and epidemiological studies on R. equi isolates.

Cite This Article

APA
Monego F, Maboni F, Krewer C, Vargas A, Costa M, Loreto E. (2009). Molecular characterization of Rhodococcus equi from horse-breeding farms by means of multiplex PCR for the vap gene family. Curr Microbiol, 58(4), 399-403. https://doi.org/10.1007/s00284-009-9370-6

Publication

Current microbiology
ISSN: 1432-0991
NlmUniqueID: 7808448
Country: United States
Language: English
Volume: 58
Issue: 4
Pages: 399-403

Researcher Affiliations

Monego, Fernanda
  • Medicina Veteirnaria Preventiva, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil. fernandamonego@hotmail.com
Maboni, Franciele
    Krewer, Cristina
      Vargas, Agueda
        Costa, Mateus
          Loreto, Elgion

            MeSH Terms

            • Actinomycetales Infections / epidemiology
            • Actinomycetales Infections / microbiology
            • Actinomycetales Infections / veterinary
            • Animals
            • Bacterial Proteins / genetics
            • Brazil / epidemiology
            • Breeding
            • Horse Diseases / epidemiology
            • Horse Diseases / microbiology
            • Horses / microbiology
            • Humans
            • Manure / microbiology
            • Membrane Glycoproteins / genetics
            • Molecular Epidemiology
            • Multigene Family / genetics
            • Polymerase Chain Reaction / methods
            • Rhodococcus equi / genetics
            • Rhodococcus equi / isolation & purification
            • Rhodococcus equi / pathogenicity
            • Soil Microbiology
            • Virulence / genetics
            • Virulence Factors / genetics

            References

            This article includes 26 references
            1. Krewer Cda C, Spricigo DA, de Avila Botton S, da Costa MM, Schrank I, de Vargas AC. Molecular characterization of Rhodococcus equi Isolates of horse breeding farms from an endemic region in South of Brazil by multiplex PCR.. Braz J Microbiol 2008 Jan;39(1):188-93.
              pubmed: 24031201doi: 10.1590/S1517-838220080001000036google scholar: lookup
            2. Halbert ND, Reitzel RA, Martens RJ, Cohen ND. Evaluation of a multiplex polymerase chain reaction assay for simultaneous detection of Rhodococcus equi and the vapA gene.. Am J Vet Res 2005 Aug;66(8):1380-5.
              pubmed: 16173481doi: 10.2460/ajvr.2005.66.1380google scholar: lookup
            3. Mizuno Y, Sato F, Sakamoto M, Yoshikawa K, Yoshida M, Shiba K, Onodera S, Matsuura R, Takai S. VapB-positive Rhodococcus equi infection in an HIV-infected patient in Japan.. J Infect Chemother 2005 Feb;11(1):37-40.
              pubmed: 15729486doi: 10.1007/s10156-004-0355-xgoogle scholar: lookup
            4. Hondalus MK. Pathogenesis and virulence of Rhodococcus equi.. Vet Microbiol 1997 Jun 16;56(3-4):257-68.
              pubmed: 9226840doi: 10.1016/s0378-1135(97)00094-1google scholar: lookup
            5. Giguère S, Hondalus MK, Yager JA, Darrah P, Mosser DM, Prescott JF. Role of the 85-kilobase plasmid and plasmid-encoded virulence-associated protein A in intracellular survival and virulence of Rhodococcus equi.. Infect Immun 1999 Jul;67(7):3548-57.
              pubmed: 10377138doi: 10.1128/IAI.67.7.3548-3557.1999google scholar: lookup
            6. 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.
              pubmed: 1937765doi: 10.1128/iai.59.11.4056-4060.1991google scholar: lookup
            7. Takai S, Hines SA, Sekizaki T, Nicholson VM, Alperin DA, Osaki M, Takamatsu D, Nakamura M, Suzuki K, Ogino N, Kakuda T, Dan H, Prescott JF. DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC 33701 and 103.. Infect Immun 2000 Dec;68(12):6840-7.
              pubmed: 11083803doi: 10.1128/IAI.68.12.6840-6847.2000google scholar: lookup
            8. Byrne BA, Prescott JF, Palmer GH, Takai S, Nicholson VM, Alperin DC, Hines SA. Virulence plasmid of Rhodococcus equi contains inducible gene family encoding secreted proteins.. Infect Immun 2001 Feb;69(2):650-6.
              pubmed: 11159951doi: 10.1128/IAI.69.2.650-656.2001google scholar: lookup
            9. 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.
              pubmed: 16957241doi: 10.1128/AEM.00495-06google scholar: lookup
            10. Bell KS, Philp JC, Christofi N, Aw DW. Identification of Rhodococcus equi using the polymerase chain reaction.. Lett Appl Microbiol 1996 Aug;23(2):72-4.
              pubmed: 8987445doi: 10.1111/j.1472-765x.1996.tb00033.xgoogle scholar: lookup
            11. Cantor GH, Byrne BA, Hines SA, Richards HM 3rd. VapA-negative Rhodococcus equi in a dog with necrotizing pyogranulomatous hepatitis, osteomyelitis, and myositis.. J Vet Diagn Invest 1998 Jul;10(3):297-300.
              pubmed: 9683085doi: 10.1177/104063879801000316google scholar: lookup
            12. Takai S, Imai Y, Fukunaga N, Uchida Y, Kamisawa K, Sasaki Y, Tsubaki S, Sekizaki T. Identification of virulence-associated antigens and plasmids in Rhodococcus equi from patients with AIDS.. J Infect Dis 1995 Nov;172(5):1306-11.
              pubmed: 7594668doi: 10.1093/infdis/172.5.1306google scholar: lookup
            13. Sydor T, von Bargen K, Becken U, Spuerck S, Nicholson VM, Prescott JF, Haas A. A mycolyl transferase mutant of Rhodococcus equi lacking capsule integrity is fully virulent.. Vet Microbiol 2008 Apr 30;128(3-4):327-41.
              pubmed: 18063488doi: 10.1016/j.vetmic.2007.10.020google scholar: lookup
            14. 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.
              pubmed: 12220812doi: 10.1016/s0378-1135(02)00157-8google scholar: lookup
            15. Takai S, Fukunaga N, Ochiai S, Sakai T, Sasaki Y, Tsubaki S. Isolation of virulent and intermediately virulent Rhodococcus equi from soil and sand on parks and yards in Japan.. J Vet Med Sci 1996 Jul;58(7):669-72.
              pubmed: 8844605doi: 10.1292/jvms.58.669google scholar: lookup
            16. Pei Y, Dupont C, Sydor T, Haas A, Prescott JF. Cholesterol oxidase (ChoE) is not important in the virulence of Rhodococcus equi.. Vet Microbiol 2006 Dec 20;118(3-4):240-6.
              pubmed: 16979852doi: 10.1016/j.vetmic.2006.08.006google scholar: lookup
            17. Jacks S, Giguère S, Prescott JF. In vivo expression of and cell-mediated immune responses to the plasmid-encoded virulence-associated proteins of Rhodococcus equi in foals.. Clin Vaccine Immunol 2007 Apr;14(4):369-74.
              pubmed: 17301216doi: 10.1128/CVI.00448-06google scholar: lookup
            18. Jain S, Bloom BR, Hondalus MK. Deletion of vapA encoding Virulence Associated Protein A attenuates the intracellular actinomycete Rhodococcus equi.. Mol Microbiol 2003 Oct;50(1):115-28.
              pubmed: 14507368doi: 10.1046/j.1365-2958.2003.03689.xgoogle scholar: lookup
            19. Staden R, Beal KF, Bonfield JK. The Staden package, 1998.. Methods Mol Biol 2000;132:115-30.
              pubmed: 10547834doi: 10.1385/1-59259-192-2:115google scholar: lookup
            20. Oldfield C, Bonella H, Renwick L, Dodson HI, Alderson G, Goodfellow M. Rapid determination of vapA/vapB genotype in Rhodococcus equi using a differential polymerase chain reaction method.. Antonie Van Leeuwenhoek 2004 May;85(4):317-26.
              pubmed: 15031644doi: 10.1023/B:ANTO.0000020383.66622.4dgoogle scholar: lookup
            21. Hondalus MK, Mosser DM. Survival and replication of Rhodococcus equi in macrophages.. Infect Immun 1994 Oct;62(10):4167-75.
              pubmed: 7927672doi: 10.1128/iai.62.10.4167-4175.1994google scholar: lookup
            22. Takai S, Anzai T, Fujita Y, Akita O, Shoda M, Tsubaki S, Wada R. Pathogenicity of Rhodococcus equi expressing a virulence-associated 20 kDa protein (VapB) in foals.. Vet Microbiol 2000 Sep 15;76(1):71-80.
              pubmed: 10925043doi: 10.1016/s0378-1135(00)00226-1google scholar: lookup
            23. 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.
              pubmed: 15563953doi: 10.1016/j.cimid.2004.07.001google scholar: lookup
            24. Takai S, Ikeda T, Sasaki Y, Watanabe Y, Ozawa T, Tsubaki S, Sekizaki T. Identification of virulent Rhodococcus equi by amplification of gene coding for 15- to 17-kilodalton antigens.. J Clin Microbiol 1995 Jun;33(6):1624-7.
              pubmed: 7650199doi: 10.1128/jcm.33.6.1624-1627.1995google scholar: lookup
            25. Napoleão F, Damasco PV, Camello TC, do Vale MD, de Andrade AF, Hirata R Jr, de Mattos-Guaraldi AL. Pyogenic liver abscess due to Rhodococcus equi in an immunocompetent host.. J Clin Microbiol 2005 Feb;43(2):1002-4.
              pubmed: 15695730doi: 10.1128/JCM.43.2.1002-1004.2005google scholar: lookup
            26. Takai S, Fukunaga N, Ochiai S, Imai Y, Sasaki Y, Tsubaki S, Sekizaki T. Identification of intermediately virulent Rhodococcus equi isolates from pigs.. J Clin Microbiol 1996 Apr;34(4):1034-7.
              pubmed: 8815079doi: 10.1128/jcm.34.4.1034-1037.1996google scholar: lookup

            Citations

            This article has been cited 5 times.
            1. Trevisani MM, Hanna ES, Oliveira AF, Cardoso SA, Roque-Barreira MC, Soares SG. Vaccination of Mice with Virulence-Associated Protein G (VapG) Antigen Confers Partial Protection against Rhodococcus equi Infection through Induced Humoral Immunity.. Front Microbiol 2017;8:857.
              doi: 10.3389/fmicb.2017.00857pubmed: 28553279google scholar: lookup
            2. Madrigal RG, Shaw SD, Witkowski LA, Sisson BE, Blodgett GP, Chaffin MK, Cohen ND. Use of Serial Quantitative PCR of the vapA Gene of Rhodococcus equi in Feces for Early Detection of R. equi Pneumonia in Foals.. J Vet Intern Med 2016 Mar-Apr;30(2):664-70.
              doi: 10.1111/jvim.13828pubmed: 26806422google scholar: lookup
            3. Shaw SD, Cohen ND, Chaffin MK, Blodgett GP, Syndergaard M, Hurych D. Estimating the Sensitivity and Specificity of Real-Time Quantitative PCR of Fecal Samples for Diagnosis of Rhodococcus equi Pneumonia in Foals.. J Vet Intern Med 2015 Nov-Dec;29(6):1712-7.
              doi: 10.1111/jvim.13631pubmed: 26436545google scholar: lookup
            4. Gressler LT, de Vargas AC, da Costa MM, Pötter L, da Silveira BP, Sangioni LA, de Avila Botton S. Genotypic and phenotypic detection of efflux pump in Rhodococcus equi.. Braz J Microbiol 2014;45(2):661-5.
              doi: 10.1590/s1517-83822014000200040pubmed: 25242956google scholar: lookup
            5. de Vargas AC, Monego F, Gressler LT, de Avila Botton S, Lazzari AM, da Costa MM, Ecco R, Ribeiro MG, Lara GH, Takai S. Bronchopneumonia in wild boar (Sus scrofa) caused by Rhodococcus equi carrying the VapB type 8 plasmid.. BMC Res Notes 2013 Mar 25;6:111.
              doi: 10.1186/1756-0500-6-111pubmed: 23531380google scholar: lookup
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