Characterization of virulence plasmid types in Rhodococcus equi isolates from foals, pigs, humans and soil in Hungary.
Abstract: Rhodococcus equi isolates (204) obtained from foals (lung abscesses, lymph nodes, nasal discharge, rectal swabs) bred in 15 studs located throughout Hungary, isolates from soil samples, lymph nodes of pigs and from lesions of human patients were examined to determine genotypic diversity of virulence-associated plasmids. Isolates were tested for the presence of 15-17 kDa virulence-associated protein antigen (VapA) and 20k Da (VapB) genes by polymerase chain reaction (PCR). Plasmid DNAs were isolated and analysed by digestion with restriction endonucleases for estimation of size and comparison of polymorphisms. Of 146 clinical isolates from foals in 15 studs, 129 (88.3%) gave positive results for the VapA gene, showing a 564 bp product of the expected size in the PCR amplification. Of the 129 clinical isolates from foals, 123 contained an 85 kb type I plasmid and the remaining six contained an 87 kb type I plasmid. Of 48 soil isolates from two horse studs, 26 (54.2%) were positive for VapA gene and contained an 85 kb type I plasmid. Of three pig isolates, one was positive for VapA gene and contained an 85 kb type I plasmid, and the remaining two were positive for the VapB gene, showing a 827 bp product of the expected size in the PCR amplification and were R. equi of intermediate virulence which contained a 95 kb type S5 plasmid. Of the seven human isolates, five were positive for VapB gene by PCR, these were R. equi of intermediate virulence, which contained a 95 kb type S5 plasmid. These results revealed that virulent R. equi strains harbouring a virulence plasmid of 85 kb type I or 87 kb type I, which have been found in clinical isolates from Europe and North and South America, are widespread in Hungary. Furthermore, same intermediately virulence plasmid type was found in both human and pig isolates.
Publication Date: 2002-09-11 PubMed ID: 12220812DOI: 10.1016/s0378-1135(02)00157-8Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research study is an investigation of the different types of virulence plasmid in Rhodococcus equi strains, isolated from various sources including foals, pigs, humans, and soil in Hungary, using polymerase chain reaction (PCR) and DNA analysis techniques.
Objective of the Research
- The objective of this research was to determine the genotypic diversity of virulence-associated plasmids in Rhodococcus equi isolates collected from different sources in Hungary.
Methodology
- 204 Rhodococcus equi isolates were obtained from different sources including foals, soil samples, lymph nodes of pigs, and lesions from human patients.
- The isolates were tested for the presence of the virulence-associated protein antigen (VapA and VapB) genes using polymerase chain reaction (PCR).
- Furthermore, the plasmid DNAs were isolated from these strains, and then analyzed by digestion with restriction endonucleases to determine the size and observe any polymorphisms.
Findings
- Of the total 146 clinical isolates from foals, 88.3% were confirmed positive for VapA gene, with PCR amplification showing a 564 bp product of the expected size.
- A majority of these isolates contained an 85 kb type I plasmid, while a small number had an 87 kb type I plasmid.
- In the case of soil isolates, 54.2% tested positive for VapA gene and also contained an 85 kb type I plasmid.
- Of the three pig isolates, one had a VapA gene and contained an 85 kb type I plasmid, while the other two were positive for the VapB gene. These two isolates showed a PCR result of 827 bp, indicating R. equi strains of intermediate virulence, and contained a 95 kb type S5 plasmid.
- Out of seven human isolates studied, five were positive for the VapB gene and had a 95 kb type S5 plasmid, suggesting R. equi strains of intermediate virulence.
Conclusions
- The results showed evidence of widespread distribution of virulent R. equi strains in Hungary, primarily those strains carrying an 85 kb or 87 kb type I virulence plasmid, similar to clinical isolates found in Europe, North, and South America.
- The study also discovered the presence of the same intermediately virulent plasmid type in both human and pig isolates.
Cite This Article
APA
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.
Vet Microbiol, 88(4), 377-384.
https://doi.org/10.1016/s0378-1135(02)00157-8 Publication
Researcher Affiliations
- Department of Microbiology and Infectious Diseases, Faculty of Veterinary Science, Szent István University, Hungária Krt. 23-25, 1143, Budapest, Hungary. makrai@novell.vmri.hu
MeSH Terms
- Actinomycetales Infections / epidemiology
- Actinomycetales Infections / microbiology
- Actinomycetales Infections / veterinary
- Animals
- Bacterial Proteins / genetics
- DNA, Bacterial / chemistry
- DNA, Bacterial / genetics
- DNA-Binding Proteins
- Genetic Variation
- Horse Diseases / epidemiology
- Horse Diseases / microbiology
- Horses
- Humans
- Hungary
- Immunocompromised Host
- Membrane Glycoproteins / genetics
- Plasmids / genetics
- Polymerase Chain Reaction / veterinary
- Prevalence
- Rhodococcus equi / classification
- Rhodococcus equi / genetics
- Rhodococcus equi / pathogenicity
- Soil Microbiology
- Swine
- Swine Diseases / epidemiology
- Swine Diseases / microbiology
- Virulence / genetics
- Virulence Factors
Citations
This article has been cited 17 times.- Löhr CV, O'Neill TW, Daw DN, Pitel MO, Schlipf JW. Pyogranulomatous enteritis and mesenteric lymphadenitis in an adult llama caused by Rhodococcus equi carrying virulence-associated protein A gene. J Vet Diagn Invest 2019 Sep;31(5):747-751.
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- Willingham-Lane JM, Berghaus LJ, Giguère S, Hondalus MK. Influence of Plasmid Type on the Replication of Rhodococcus equi in Host Macrophages. mSphere 2016 Sep-Oct;1(5).
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- Valero-Rello A, Hapeshi A, Anastasi E, Alvarez S, Scortti M, Meijer WG, MacArthur I, Vázquez-Boland JA. An Invertron-Like Linear Plasmid Mediates Intracellular Survival and Virulence in Bovine Isolates of Rhodococcus equi. Infect Immun 2015 Jul;83(7):2725-37.
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- Poolkhet C, Chumsing S, Wajjwalku W, Minato C, Otsu Y, Takai S. Plasmid Profiles and Prevalence of Intermediately Virulent Rhodococcus equi from Pigs in Nakhonpathom Province, Thailand: Identification of a New Variant of the 70-kb Virulence Plasmid, Type 18. Vet Med Int 2010;2010:491624.
- Monego F, Maboni F, Krewer C, Vargas A, Costa M, Loreto E. Molecular characterization of Rhodococcus equi from horse-breeding farms by means of multiplex PCR for the vap gene family. Curr Microbiol 2009 Apr;58(4):399-403.
- Rodríguez-Lázaro D, Lewis DA, Ocampo-Sosa AA, Fogarty U, Makrai L, Navas J, Scortti M, Hernández M, Vázquez-Boland JA. Internally controlled real-time PCR method for quantitative species-specific detection and vapA genotyping of Rhodococcus equi. Appl Environ Microbiol 2006 Jun;72(6):4256-63.
- Makrai L, Takayama S, Dénes B, Hajtós I, Sasaki Y, Kakuda T, Tsubaki S, Major A, Fodor L, Varga J, Takai S. Characterization of virulence plasmids and serotyping of rhodococcus equi isolates from submaxillary lymph nodes of pigs in Hungary. J Clin Microbiol 2005 Mar;43(3):1246-50.
- Matsuoka M, Kobayashi A, Matsuoka M, Honda Y, Hashimoto R, Sasaki Y, Kakuda T, Suzuki Y, Takai S. Isolation of vapB-positive Rhodococcus equi from submaxillary lymph nodes with or without granulomatous lesions in growing-finishing pigs in Japan. J Vet Med Sci 2024 Jun 1;86(6):600-605.
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