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Frontiers in microbiology2025; 16; 1635639; doi: 10.3389/fmicb.2025.1635639

Isolation, identification and comparative genomic analysis of Lactobacillus salivarius from Mongolian horse vagina.

Abstract: Reproductive health in mares is pivotal for the sustainability of the equine industry, yet vaginal microbiota dysbiosis remains an underrecognized contributor to infections such as endometritis and bacterial vaginosis. While spp. dominate healthy vaginal ecosystems in humans and livestock, their role in equine reproductive health, particularly in resilient breeds like Mongolian mares, is poorly understood. This study aimed to isolate and characterize a novel strain from the vaginal microbiota of healthy Mongolian mares and evaluate its probiotic potential for mitigating equine reproductive disorders. Unassigned: A polyphasic approach integrating phenotypic, biochemical, and 16S rRNA gene sequencing was employed to isolate and identify a novel strain (Y20) from the vaginal microbiota of healthy Mongolian mares. The probiotic potential of Y20 was assessed through assays, including tolerance to low pH (pH 2.5) and bile salts (0.3%), antagonistic activity against equine pathogens (, , ), and antioxidant capacity (DPPH radical scavenging assay). Whole-genome sequencing (1.74 Mb, 33.01% GC content) was performed to analyze genes related to carbohydrate metabolism, adhesion factors, bacteriocin biosynthesis, and stress response pathways. Comparative genomics was used to explore phylogenetic relationships and genomic adaptations for vaginal colonization. Unassigned: Y20 demonstrated robust tolerance to low pH and bile salts, potent antagonistic activity against key equine pathogens, and significant antioxidant capacity (82.4% DPPH radical scavenging). Genomic analysis revealed genes encoding carbohydrate metabolism, adhesion factors, bacteriocin biosynthesis (including a novel putative bacteriocin cluster), and stress response pathways. Comparative genomics confirmed Y20's close phylogenetic relationship with horse-derived strains and identified unique genomic adaptations for vaginal colonization. Unassigned: These findings identify Y20 as a promising candidate probiotic for mitigating equine reproductive disorders. Its multifaceted probiotic properties, including pathogen inhibition, antioxidant activity, and genomic adaptations for vaginal colonization, suggest potential applications in sustainable equine health management. This study advances microbiome-based strategies as viable alternatives to antibiotics, offering new avenues for improving reproductive health in mares.
Copyright © 2025 Zhao, Liu, Tao, Cao, Lin, He, Fang, Yun, Du, Su, Bao, Bai, Zhang and Dugarjaviin.
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Publication Date: 2025-07-31 PubMed ID: 40822386PubMed Central: PMC12350343DOI: 10.3389/fmicb.2025.1635639Google 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.

Overview

  • This study isolated and identified a novel strain of Lactobacillus salivarius (Y20) from the vaginal microbiota of healthy Mongolian mares.
  • The research evaluated the probiotic potential of this strain through laboratory assays and whole-genome analysis, showing promising properties for improving equine reproductive health.

Background and Importance

  • Reproductive health in mares is critical for the equine industry’s sustainability.
  • Disruptions in vaginal microbiota, termed dysbiosis, contribute to infections like endometritis and bacterial vaginosis.
  • Lactobacillus species dominate vaginal microbiota in humans and many livestock, playing protective roles, but their role in horses—especially resilient breeds like Mongolian mares—is not well understood.
  • The study addresses this knowledge gap by isolating and characterizing a vaginal Lactobacillus salivarius strain from healthy Mongolian mares.

Objectives

  • Isolate and identify a novel Lactobacillus salivarius strain from healthy Mongolian mare vaginal microbiota.
  • Evaluate probiotic properties relevant to equine reproductive health, including survival in harsh conditions, pathogen inhibition, and antioxidant activity.
  • Conduct whole-genome sequencing to understand genetic factors associated with probiotic functions and vaginal colonization.
  • Perform comparative genomic analysis to investigate phylogenetic relationships with other horse-derived L. salivarius strains and adaptations that enable vaginal colonization.

Methods

  • Isolation and Identification:
    • A polyphasic approach combining phenotypic (observable characteristics), biochemical tests, and 16S rRNA gene sequencing was used to identify the strain Y20 as Lactobacillus salivarius.
    • Sampled from healthy Mongolian mare vaginal microbiota.
  • Probiotic Potential Assessment:
    • Survival assays tested strain tolerance to low pH (2.5) and bile salts (0.3%), mimicking vaginal and digestive stress conditions.
    • Antagonistic activity was assessed against equine pathogens including species from genera Streptococcus, Escherichia, and Pseudomonas.
    • Antioxidant capacity measured using DPPH radical scavenging assay, evaluating the strain’s ability to neutralize free radicals.
  • Genomic Analysis:
    • Whole-genome sequencing produced a draft genome of 1.74 Mb with 33.01% GC content.
    • Genome annotated for genes involved in carbohydrate metabolism, adhesion factors (important for colonization), bacteriocin biosynthesis (antimicrobial peptides), and stress response mechanisms.
  • Comparative Genomics:
    • Phylogenetic analysis positioned Y20 close to other horse-derived L. salivarius strains.
    • Identified unique genomic features suggesting adaptations for survival and colonization in the vaginal environment.

Key Findings

  • Y20 exhibited strong tolerance to challenging conditions similar to those in the equine vaginal environment (low pH and bile salts).
  • The strain demonstrated potent antagonistic activity against major equine pathogens, implying a protective role against infections.
  • Showed significant antioxidant activity, scavenging 82.4% of DPPH radicals, which could reduce oxidative stress in the reproductive tract.
  • Genomic content included:
    • Genes for carbohydrate metabolism, enabling utilization of available sugars for growth.
    • Adhesion factors to facilitate colonization and persistence in the vaginal mucosa.
    • Bacteriocin biosynthesis clusters, including a novel putative cluster, suggesting capability to produce antimicrobial compounds.
    • Stress response genes supporting survival in hostile environments.
  • Phylogenetic and comparative analysis confirmed close relation to other equine-derived L. salivarius strains but also highlighted unique adaptations enabling vaginal niche colonization.

Implications and Applications

  • The isolated strain Y20 is a promising probiotic candidate to help mitigate equine reproductive disorders by:
    • Inhibiting pathogen growth and colonization.
    • Reducing oxidative stress through antioxidant activity.
    • Stably colonizing the vaginal environment due to specialized adhesion mechanisms.
  • This provides a microbiome-based alternative to antibiotics, important for combating antimicrobial resistance.
  • The findings support development of sustainable equine health management strategies focusing on natural microbial interventions.
  • This research enhances understanding of the equine vaginal microbiome and its role in reproductive health, which has been less studied compared to other species.

Conclusion

  • The study successfully isolated and thoroughly characterized a novel Lactobacillus salivarius strain with strong probiotic properties from Mongolian mare vaginal microbiota.
  • The integrated phenotypic, biochemical, and genomic evidence supports Y20’s potential as a beneficial microbe for improving reproductive health and preventing infections in horses.
  • This work opens new avenues for using probiotics to enhance mare fertility and overall animal wellbeing in the equine industry.

Cite This Article

APA
Zhao Y, Liu Y, Tao J, Cao J, Lin Y, He Q, Fang X, Yun S, Du M, Su S, Bao T, Bai D, Zhang X, Dugarjaviin M. (2025). Isolation, identification and comparative genomic analysis of Lactobacillus salivarius from Mongolian horse vagina. Front Microbiol, 16, 1635639. https://doi.org/10.3389/fmicb.2025.1635639

Publication

Frontiers in microbiology
ISSN: 1664-302X
NlmUniqueID: 101548977
Country: Switzerland
Language: English
Volume: 16
Pages: 1635639
PII: 1635639

Researcher Affiliations

Zhao, Yiping
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Liu, Yuanyi
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Tao, Jinshan
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Cao, Jialong
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Lin, Yanan
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
He, Qianqian
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Fang, Xinlan
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Yun, Siqin
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Du, Ming
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Su, Shaofeng
  • Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot, China.
Bao, Tugeqin
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Bai, Dongyi
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Zhang, Xinzhuang
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Dugarjaviin, Manglai
  • Key Laboratory of Equus Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Hohhot, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot, China.
  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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