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BMC veterinary research2024; 20(1); 362; doi: 10.1186/s12917-024-04167-9

Whole-genome sequencing and pathogenicity analysis of Rhodococcus equi isolated in horses.

Abstract: Rhodococcus equi (R. equi) is a Gram-positive zoonotic pathogen that frequently leads to illness and death in young horses (foals). This study presents the complete genome sequence of R. equi strain BJ13, which was isolated from a thoroughbred racehorse breeding farm in Beijing, China. Results: The BJ13 genome has a length of 5.30 Mb and consists of a complete chromosome and a plasmid measuring 5.22 Mb and 0.08 Mb, respectively. We predicted 4,929 coding gene open reading frames, along with 52 tRNAs and 12 rRNAs. Through analysis of mobile genetic elements, we identified 6 gene islands and 1 prophage gene. Pathogenic system analysis predicted the presence of 418 virulence factors and 225 drug resistance genes. Secretion system analysis revealed the prediction of 297 secreted proteins and 1,106 transmembrane proteins. BJ13 exhibits genomic features, virulence-associated genes, potential drug resistance, and a virulence plasmid structure that may contribute to the evolution of its pathogenicity. Lastly, the pathogenicity of the isolated strain was assessed through animal experiments, which resulted in inflammatory reactions or damage in the lungs, liver, and spleen of mice. Moreover, by the 7th day post-infection, the mortality rate of the mice reached 50.0%, indicating complex immune regulatory mechanisms, including overexpression of IL-10 and increased production of pro-inflammatory cytokines like TNF-α. These findings validate the strong pathogenicity of the isolated strain and provide insights for studying the pathogenic mechanisms of Rhodococcus equi infection. Conclusions: The complete genome sequence of R. equi strain BJ13 provides valuable insights into its genomic characteristics, virulence potential, drug resistance, and secretion systems. The strong pathogenicity observed in animal experiments underscores the need for further investigation into the pathogenic mechanisms of R. equi infection.
Publication Date: 2024-08-12 PubMed ID: 39129003PubMed Central: PMC11318318DOI: 10.1186/s12917-024-04167-9Google Scholar: Lookup
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  • Journal Article

Summary

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Overview

  • This research focuses on the whole-genome sequencing and pathogenicity analysis of Rhodococcus equi (R. equi) strain BJ13, a bacterium isolated from horses.
  • The study identifies genomic features, virulence factors, and drug resistance genes of the strain and confirms its strong pathogenic effects through animal experiments.

Background and Importance

  • Rhodococcus equi: A Gram-positive zoonotic pathogen known to cause severe illness in young horses (foals), leading to significant morbidity and mortality.
  • Zoonotic nature: R. equi can potentially infect humans, highlighting the importance of understanding its pathogenic mechanisms.
  • Objective: Detailed genome sequencing of R. equi strain BJ13, isolated from a thoroughbred racehorse breeding farm in China, to understand its genetic makeup and virulence traits.

Genome Sequencing and Analysis

  • Genome Length: The BJ13 strain’s genome is 5.30 million base pairs (Mb) in size, comprising a complete chromosome (5.22 Mb) and a plasmid (0.08 Mb).
  • Genetic Content:
    • 4,929 predicted coding gene open reading frames (ORFs)
    • 52 transfer RNA (tRNA) genes
    • 12 ribosomal RNA (rRNA) genes
  • Mobile Genetic Elements: Identification of 6 gene islands and 1 prophage gene within the genome, which could contribute to horizontal gene transfer and variability in virulence.

Virulence Factors and Drug Resistance Genes

  • Virulence Factors: The analysis predicted 418 genes related to virulence, indicating multiple mechanisms by which the bacterium can infect and damage host tissues.
  • Drug Resistance: A total of 225 genes potentially linked to resistance against various antimicrobial agents were identified, highlighting challenges in treatment.
  • Virulence Plasmid: The plasmid present in BJ13 strain includes genes that contribute to pathogenicity, supporting the ability of the strain to cause disease.

Secretion Systems and Protein Prediction

  • Secreted Proteins: 297 secreted proteins were predicted, which could be involved in host interaction and immune evasion.
  • Transmembrane Proteins: 1,106 proteins predicted to span cell membranes, likely playing roles in nutrient transport, signaling, or immune system interactions.

Pathogenicity Assessment Through Animal Experiments

  • Experimental Model: Infection of mice to study the pathogenic effects of BJ13 strain.
  • Observed Effects:
    • Inflammatory responses and tissue damage noted in lungs, liver, and spleen.
    • By day 7 post-infection, 50% mortality rate in mice, indicating high virulence.
  • Immune Response: Complex immune regulation observed, including:
    • Overexpression of IL-10, an anti-inflammatory cytokine.
    • Increased production of pro-inflammatory cytokines such as TNF-α.

Conclusions and Implications

  • The complete genome sequencing of R. equi BJ13 sheds light on its genetic basis for virulence, drug resistance, and secretory capacity.
  • The strong pathogenicity demonstrated in animal models underlines the need for deeper investigation into mechanisms of infection and host immune response interaction.
  • Insights gained may aid in developing targeted interventions or therapeutics to manage R. equi infections in horses and possibly prevent zoonotic transmission.

Cite This Article

APA
Hu B, Gao S, Zhang H, Li Q, Li G, Zhang S, Xing Y, Huang Y, Han S, Tian Y, Zhang W, He H. (2024). Whole-genome sequencing and pathogenicity analysis of Rhodococcus equi isolated in horses. BMC Vet Res, 20(1), 362. https://doi.org/10.1186/s12917-024-04167-9

Publication

ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 20
Issue: 1
Pages: 362
PII: 362

Researcher Affiliations

Hu, Bin
  • College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, China.
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • University of Chinese Academy of Sciences, Beijing, China.
Gao, Sichao
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • Anhui University of Science and Technology, Huainan, China.
Zhang, Hao
  • University of Chinese Academy of Sciences, Beijing, China.
Li, Qiaoqiao
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • Anhui University of Science and Technology, Huainan, China.
Li, Gaojian
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • University of Chinese Academy of Sciences, Beijing, China.
Zhang, Shuairan
  • College of Shenyang Institute of Technology, Shenyang, Liaoning, China.
Xing, Yanan
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • University of Chinese Academy of Sciences, Beijing, China.
Huang, Yanyi
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • University of Chinese Academy of Sciences, Beijing, China.
Han, Shuyi
  • CAS Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • University of Chinese Academy of Sciences, Beijing, China.
Tian, Ying
  • Beijing Wildlife Rescue and Rehabilitation Center, Beijing, China.
Zhang, Wei
  • Beijing Wildlife Rescue and Rehabilitation Center, Beijing, China.
He, Hongxuan
  • College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, China. hehx@ioz.ac.cn.

MeSH Terms

  • Rhodococcus equi / pathogenicity
  • Rhodococcus equi / genetics
  • Animals
  • Horses
  • Horse Diseases / microbiology
  • Actinomycetales Infections / veterinary
  • Actinomycetales Infections / microbiology
  • Whole Genome Sequencing
  • Virulence / genetics
  • Genome, Bacterial
  • Mice
  • Virulence Factors / genetics
  • Female

Grant Funding

  • No. 2022YFC2601602 / the National Key Research and Development Program of China
  • No. 2022YFC2601602 / the National Key Research and Development Program of China
  • No. 2022YFC2601602 / the National Key Research and Development Program of China
  • No. 32090023 / the Major Program of National Natural Science Foundation of China
  • No. 32090023 / the Major Program of National Natural Science Foundation of China
  • No. 32090023 / the Major Program of National Natural Science Foundation of China

Conflict of Interest Statement

The authors declare no competing interests.

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