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Home / Equine Research Bank / Microbiol Immunol / Rhodococcus equiU19 strain harbors a nonmobilizable virulenc...
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

References

This article includes 30 references
  1. Takai S. Epidemiology of Rhodococcus equi infections: a review.. Vet Microbiol 1997;56:167-76.
  2. Takai S, Madarame H, Sasaki Y, Suzuki Y, Kakuda T. Rhodococcus equi infections in domestic animals, companion animals, and wildlife.. JVAM 2021;74:695-706.
  3. Suzuki Y, Kubota H, Madarame H. Pathogenicity and genomic features of vapN-harboring Rhodococcus equi isolated from human patients.. Int J Med Microbiol 2021;311:151519.
  4. Suzuki Y, Takahashi K, Takase F. Serological epidemiological surveillance for vapN-harboring Rhodococcus equi infection in goats.. Comp Immunol Microbiol Infect Dis 2020;73:101540.
  5. Takai S, Sawada N, Nakayama Y. Reinvestigation of the virulence of Rhodococcus equi isolates from patients with and without AIDS.. Lett Appl Microbiol 2020;71:679-83.
  6. MacArthur I, Anastasi E, Alvarez S, Scortti M, Vázquez-Boland JA. Comparative genomics of Rhodococcus equi virulence plasmids indicates host-driven evolution of the vap pathogenicity Island.. Genome Biol Evol 2017;9:1241-47.
  7. Valero-Rello A, Hapeshi A, Anastasi E. An invertron-like linear plasmid mediates intracellular survival and virulence in bovine isolates of Rhodococcus equi.. Infect Immun 2015;83:2725-37.
  8. Vázquez-Boland JA, Meijer WG. The pathogenic actinobacterium Rhodococcus equi: what's in a name?. Mol Microbiol 2019;112:1-15.
  9. von Bargen K, Scraba M, Krämer I. Virulence-associated protein A from Rhodococcus equi is an intercompartmental pH-neutralising virulence factor.. Cell Microbiol 2019;21:e12958.
  10. Letek M, Ocampo-Sosa AA, Sanders M. Evolution of the Rhodococcus equi vap pathogenicity island seen through comparison of host-associated vapA and vapB virulence plasmids.. J Bacteriol 2008;190:5797-805.
  11. Takai S, Hines SA, Sekizaki T. DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC 33701 and 103.. Infect Immun 2000;68:6840-47.
  12. Takai S, Sugawara T, Watanabe Y, Sasaki Y, Tsubaki S, Sekizaki T. Effect of growth temperature on maintenance of virulent Rhodococcus equi.. Vet Microbiol 1994;39:187-92.
  13. Tripathi VN, Harding WC, Willingham-Lane JM, Hondalus MK. Conjugal transfer of a virulence plasmid in the opportunistic intracellular actinomycete Rhodococcus equi.. J Bacteriol 2012;194:6790-801.
  14. Sekine M, Tanikawa S, Omata S. Sequence analysis of three plasmids harboured in Rhodococcus erythropolis strain PR4.. Environ Microbiol 2006;8:334-46.
  15. Honda K, Imura M, Okano K, Omasa T, Kato J, Ohtake H. Identification of the replication region of a 111-kb circular plasmid from Rhodococcus opacus B-4 by λ Red recombination-based deletion analysis.. Biosci Biotechnol Biochem 2012;76:1758-64.
  16. Smillie C, Garcillán-Barcia MP, Francia MV, Rocha EP, de la Cruz F. Mobility of plasmids.. Microbiol Mol Biol Rev 2010;74:434-52.
  17. Garcillán-Barcia MP, Francia MV, de la Cruz F. The diversity of conjugative relaxases and its application in plasmid classification.. FEMS Microbiol Rev 2009;33:657-87.
  18. 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;59:4056-60.
  19. Takai S, Sasaki Y, Ikeda T, Uchida Y, Tsubaki S, Sekizaki T. Virulence of Rhodococcus equi isolates from patients with and without AIDS.. J Clin Microbiol 1994;32:457-60.
  20. Takai S, Tharavichitkul P, Takarn P. Molecular epidemiology of Rhodococcus equi of intermediate virulence isolated from patients with and without acquired immune deficiency syndrome in Chiang Mai, Thailand.. J Infect Dis 2003;188:1717-23.
  21. Suzuki Y, Takahashi K, Takase F. Serological epidemiological surveillance for vapN-harboring Rhodococcus equi infection in goats.. Comp Immunol Microbiol Infect Dis 2020;73:101540.
  22. Takai S, Iie M, Kobayashi C. Monoclonal antibody specific to virulence-associated 15- to 17-kilodalton antigens of Rhodococcus equi.. J Clin Microbiol 1993;31:2780-82.
  23. Sangkanjanavanich N, Kawai M, Kakuda T, Takai S. Rescue of an intracellular avirulent Rhodococcus equi replication defect by the extracellular addition of virulence-associated protein A.. J Vet Med Sci 2017;79:1323-26.
  24. Takai S, Ikeda T, Sasaki Y. Identification of virulent Rhodococcus equi by amplification of gene coding for 15- to 17-kilodalton antigens.. J Clin Microbiol 1995;33:1624-27.
  25. Wick RR, Judd LM, Gorrie CL, Holt KE. Unicycler: resolving bacterial genome assemblies from short and long sequencing reads.. PLoS Comput Biol 2017;13:e1005595.
  26. Andreu N, Zelmer A, Fletcher T. Optimisation of bioluminescent reporters for use with mycobacteria.. PLoS One 2010;5:e10777.
  27. 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;14:243.
  28. Huber L, Giguere S, Slovis NM. The novel and transferable erm(51) gene confers macrolides, lincosamides and streptogramins B (MLSB) resistance to clonal Rhodococcus equi in the environment.. Environ Microbiol 2020;22:2858-69.
  29. Takai S, Fukunaga N, Kamisawa K, Imai Y, Sasaki Y, Tsubaki S. Expression of virulence-associated antigens of Rhodococcus equi is regulated by temperature and pH.. Microbiol Immunol 1996;40:591-4.
  30. Matsui K, Shintani M, Yamane H, Nojiri H. Genetic elements and environmental factors involved in the plasmid conjugation.. J Environ Biotechnol 2011;11:69-75.

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