Two new variants of the Rhodococcus equi virulence plasmid, 90 kb type III and type IV, recovered from a foal in Japan.
Abstract: This report describes the discovery of two new virulence plasmid types from a crossbred foal with Rhodococcus equi pneumonia in Kumamoto died with severe R. equi pneumonia and ulcerative enteritis. R. equi was isolated in large numbers and isolates from the foal were investigated for the presence of virulence-associated 15-17 kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and by gene coding for VapA by PCR. Plasmid DNAs extracted from the isolates were digested with restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII. The digestion patterns that resulted divided the plasmids of these isolates into two closely related types. The digestion patterns were then compared with eight representative virulence plasmid types (85 kb types I, II, III and IV, 87 kb types I and II, 90 kb types I and II), which have already been reported. None of the EcoRI and EcoT22I digestion patterns of the eight representative plasmids matched those of the two plasmid types. We tentatively designated these new plasmid types as 90 kb type III and type IV, since HindIII and BamHI digestion patterns of the two plasmid types were identical with those of a 90 kb type I plasmid. This study, demonstrated that there are at least 10 distinct but closely related plasmids present in isolates from horses in the world.
Publication Date: 2001-08-17 PubMed ID: 11506930DOI: 10.1016/s0378-1135(01)00398-4Google Scholar: Lookup
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- Case Reports
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research study describes the identification of two new and closely-related plasmid types, which contribute to the virulence or disease-causing ability of the bacterium Rhodococcus equi, in a sick foal in Japan.
Details of the Study
- The foal, from which the bacterium was isolated, was suffering from a severe case of Rhodococcus equi pneumonia and ulcerative enteritis.
- The focus of the study was on identifying the presence of virulence-associated antigens (VapA) which play a key role in the pathogenicity of R. equi. Identifying different variations of the VapA gene can help understand the diversity and adaptability of this bacterium to host organisms.
Methods Employed
- The investigators used colony blotting and a special kind of stain (monoclonal antibody 10G5) to identify the presence of the VapA antigens in the bacterium.
- The detection of the VapA coding gene was facilitated by Polymerase Chain Reaction (PCR), a molecular biology technique used to amplify and thus detect targeted DNA sequences.
- Further exploration of the virulence-associated plasmids was carried out by using certain enzymes (restriction endonucleases) to break down the DNA at specific points and study the resultant digestion patterns.
Findings and Conclusions
- The digestion patterns identified were compared with those of eight other known plasmid types, previously reported.
- This comparison revealed that the plasmids extracted from the foal did not match any of the known types, therefore indicating the discovery of two new types. These were tentatively named as 90 kb type III and type IV.
- The results of this study add to our understanding of R. equi by highlighting the existence of at least 10 distinct, yet closely-related, plasmids in this bacterium within horse populations worldwide.
Cite This Article
APA
Takai S, Murata N, Kudo R, Narematsu N, Kakuda T, Sasaki Y, Tsubaki S.
(2001).
Two new variants of the Rhodococcus equi virulence plasmid, 90 kb type III and type IV, recovered from a foal in Japan.
Vet Microbiol, 82(4), 373-381.
https://doi.org/10.1016/s0378-1135(01)00398-4 Publication
Researcher Affiliations
- Department of Animal Hygiene, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, 034-8628, Aomori, Japan. takai@vmas.kitasato-u.ac.jp
MeSH Terms
- Actinomycetales Infections / microbiology
- Actinomycetales Infections / veterinary
- Animals
- Animals, Newborn
- DNA, Bacterial / analysis
- Enteritis / microbiology
- Enteritis / veterinary
- Fatal Outcome
- Horse Diseases / microbiology
- Horses
- Immunohistochemistry / veterinary
- Male
- Plasmids / classification
- Plasmids / genetics
- Pneumonia, Bacterial / microbiology
- Pneumonia, Bacterial / veterinary
- Polymerase Chain Reaction
- Restriction Mapping
- Rhodococcus equi / genetics
- Rhodococcus equi / pathogenicity
- Virulence
Citations
This article has been cited 5 times.- Ganbaatar O, Ganzorig S, Tseren-Ochir EO, Suzuki Y, Takai S. Isolation of vapA-positive Rhodococcus equi from soil and fecal samples in Mongolia. J Vet Med Sci 2025 Oct 1;87(10):1112-1115.
- Nakagawa R, Moki H, Hayashi K, Ooniwa K, Tokuyama K, Kakuda T, Yoshioka K, Takai S. A case report on disseminated Rhodococcus equi infection in a Japanese black heifer. J Vet Med Sci 2018 May 18;80(5):819-822.
- Witkowski L, Rzewuska M, Takai S, Chrobak-Chmiel D, Kizerwetter-Świda M, Feret M, Gawryś M, Witkowski M, Kita J. Molecular characterization of Rhodococcus equi isolates from horses in Poland: pVapA characteristics and plasmid new variant, 85-kb type V. BMC Vet Res 2017 Jan 26;13(1):35.
- Kakuda T, Hirota T, Takeuchi T, Hagiuda H, Miyazaki S, Takai S. VirS, an OmpR/PhoB subfamily response regulator, is required for activation of vapA gene expression in Rhodococcus equi. BMC Microbiol 2014 Oct 3;14:243.
- 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.
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