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Microbiology and immunology2022; 66(6); 307-316; doi: 10.1111/1348-0421.12975

Rhodococcus equiU19 strain harbors a nonmobilizable virulence plasmid.

Abstract: Rhodococcus equiis the causative agent of pyogenic pneumonia in foals, and a virulence-associated protein A (VapA) encoded on the pVAPA virulence plasmid is important for its pathogenicity. In this study, we analyzed the virulence of R. equi strain U19, originally isolated in the Netherlands in 1997 and the genetic characteristics of the pVAPA_U19 plasmid. U19 expressed VapA that was regulated by temperature and pH and underwent significant intracellular proliferation in macrophages. The restriction fragment length polymorphism of pVAPA_U19 digested with EcoRI was similar to that of pREAT701 (85 kb Type I) harbored by R. equi ATCC33701, although the band pattern at 10-20 kb differed. Whole-genome sequencing showed that pVAPA_U19 was 51,684 bp in length and that the vapA pathogenicity island region and the replication/participation were almost identical to those in pREAT701. By contrast, the open reading frames (ORF26-ORF45) genes of pREAT701 (approximately 29,000 bp) were absent from pVAPA_U19. In this lacking region, mobility (MOB) genes, such as relaxase, which allow conjugative DNA processing, and the mating pair formation (MPF) genes, which are a form of the Type IV secretion system and provide the mating channel, were present. Coculture between U19 and five different recipient strains (two plasmid-cured strains and three cryptic plasmid-harboring strains) demonstrated that pVAPA_U19 could not support conjugation. Therefore, pVAPA_U19 does not differ significantly from the previously reported pVAPA in terms of virulence and plasmid replication and maintenance but is a nonmobilizable plasmid unable to cause conjugation because of the absence of genes related to MOB and MPF.
© 2022 The Societies and John Wiley & Sons Australia, Ltd.
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Publication Date: 2022-04-13 PubMed ID: 35274358DOI: 10.1111/1348-0421.12975Google 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 research team studied a strain of Rhodococcus equi, called U19, to investigate its virulence and genetic characteristics. They found that, unlike other similar strains, the U19 strain does not contain genes that allow for DNA conjugation, the process of DNA transfer during bacterial mating.

Analysis of R. equi U19 Strain’s Virulence

  • The researchers focused on strains of bacteria known as R. equi, particularly a strain called U19. These bacteria can cause infection and illness in foals, young horses.
  • The researchers were particularly interested in a protein produced by this bacteria, VapA. This protein is encoded on the pVAPA virulence plasmid and is critical for the bacteria’s ability to cause disease.
  • The U19 strain was found to express the VapA protein and underwent significant intracellular proliferation within macrophages, large white blood cells that consume bacteria and other pathogens.
  • The expression of the VapA protein was regulated by changes in temperature and acidity (pH).

Genetic Analysis of R. equi U19 Strain

  • The researchers conducted a genetic analysis of the U19 strain. This involved sequencing the whole genome of the bacteria and closely examining the pVAPA_U19 plasmid.
  • The pVAPA_U19 plasmid was found to be 51,684 base pairs in length and the vapA pathogenicity island region and the replication/participation were almost identical to those in pREAT701, another plasmid harbored by R. equi ATCC33701.
  • There were differences in certain parts of the plasmid, particularly in the area concerning open reading frames (ORFs) genes. In fact, approximately 29,000 base pairs in this section (from ORF26 to ORF45) were found to be missing in pVAPA_U19. This missing region is where genes allowing DNA conjugation (MOB and MPF) are typically located.

Non-mobilizable Nature of pVAPA_U19 Plasmid

  • The absence of these MOB and MPF genes suggests that pVAPA_U19 is a nonmobilizable plasmid. Conjugation, a process that allows DNA to be transferred from one bacterium to another, was found to be unsupported by pVAPA_U19.
  • This finding was supported by coculture experiments, which involved growing the U19 strain with five other strains that also lack these genes.
  • The overall implication of these findings is that while pVAPA_U19 doesn’t differ significantly from other similar plasmids in terms of virulence, it lacks the potential to transfer its DNA to other bacterial strains through conjugation.

Cite This Article

APA
Suzuki Y, Takai S, Kubota H, Hasegawa N, Ito S, Yabuuchi Y, Sasaki Y, van Duijkeren E, Kakuda T. (2022). Rhodococcus equiU19 strain harbors a nonmobilizable virulence plasmid. Microbiol Immunol, 66(6), 307-316. https://doi.org/10.1111/1348-0421.12975

Publication

Microbiology and immunology
ISSN: 1348-0421
NlmUniqueID: 7703966
Country: Australia
Language: English
Volume: 66
Issue: 6
Pages: 307-316

Researcher Affiliations

Suzuki, Yasunori
  • Laboratory of Animal Hygiene, Kitasato University School of Veterinary Medicine, Aomori, Japan.
Takai, Shinji
  • Laboratory of Animal Hygiene, Kitasato University School of Veterinary Medicine, Aomori, Japan.
Kubota, Hiroaki
  • Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Tokyo, Japan.
Hasegawa, Noeru
  • Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Tokyo, Japan.
Ito, Shino
  • Laboratory of Animal Hygiene, Kitasato University School of Veterinary Medicine, Aomori, Japan.
Yabuuchi, Yoshino
  • Laboratory of Animal Hygiene, Kitasato University School of Veterinary Medicine, Aomori, Japan.
Sasaki, Yukako
  • Laboratory of Animal Hygiene, Kitasato University School of Veterinary Medicine, Aomori, Japan.
van Duijkeren, Engeline
  • Center for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands.
Kakuda, Tsutomu
  • Laboratory of Animal Hygiene, Kitasato University School of Veterinary Medicine, Aomori, Japan.

MeSH Terms

  • Animals
  • Bacterial Proteins / genetics
  • Horse Diseases
  • Horses / genetics
  • Plasmids / genetics
  • Rhodococcus / genetics
  • Rhodococcus equi / genetics
  • Virulence / genetics
  • Virulence Factors / genetics

Grant Funding

  • JP21K14975 / Japan Society for the Promotion of Science

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Citations

This article has been cited 1 times.
  1. 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.
    doi: 10.1292/jvms.25-0267pubmed: 40754416google scholar: lookup
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