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Home / Equine Research Bank / Microbiol Spectr / Clostridioides difficile in equidae necropsied in Northweste...
Microbiology spectrum2025; 14(2); e0216525; doi: 10.1128/spectrum.02165-25

Clostridioides difficile in equidae necropsied in Northwestern France, between 2019 and 2021.

Abstract: an anaerobic, spore-forming enteropathogen, is less studied in animals than in humans despite its importance in One Health. We evaluated occurrence, diversity, circulation, and virulence in French Equidae ( = 100) after their necropsy in northwestern France, from 2019 to 2021. We systematically recovered all cecal contents and any watery intestinal contents. We isolated strains, determined their toxin gene profile by PCR, and established their PCR-ribotype according to the WEBRIBO database. We also performed free toxin detection. Twenty-seven Equidae were positive for and 20 had a toxigenic strain, including one animal co-colonized by a non-toxigenic strain. Toxigenic isolates belonged to eight already described ribotypes: (i) 078 and 126 (toxin genes of wild-type size: ), (ii) 005, 012, 020, AI-53, and FR227 (), and (iii) 017 (). Non-toxigenic isolates were of ribotypes 009, 035, and 439. Ribotypes 017 and 009 were predominant ( = 5). In two cases, Equidae of the same premises shared the same ribotype, either 020 or 009. Free toxins were detected in four animals: they displayed signs of diarrhea and a of ribotype 126 ( = 1) or 017 ( = 3) as the only detected enteropathogen, suggesting a infection (CDI). Three of them had received antibiotics. Two had died from an entero-toxic infection, for which ribotype 017 was the only identified cause. French Equidae displayed common pathogenic and ribotype 017 was highly virulent. These findings are of concern from a One Health perspective.IMPORTANCE, a major enteropathogen widely disseminated in the environment, is a One Health issue. Animals are raising concern as human contamination sources. Equidae are in close contact with humans and also develop -antibiotic and healthcare-associated CDIs. The systematic survey of Equidae necropsied from 2019 to 2021 in the leading horse breeding region in France revealed that 20% harbored pathogenic strains. These belonged to clinically important ribotypes, raising the possibility of cross-species, zoonotic or anthropo-zoonotic transmission. Free toxins, which are rarely tested in animals, were detected in four animals with signs of diarrhea and a toxigenic as the only identified enteropathogen, suggesting CDI. In two of them, ribotype 017 was the only identified cause of entero-toxic disease and death. French Equidae could play a role in the dissemination of pathogenic and notably ribotype 017. They should be surveilled carefully from a One Health perspective.
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Publication Date: 2025-12-30 PubMed ID: 41467783PubMed Central: PMC12889072DOI: 10.1128/spectrum.02165-25Google Scholar: Lookup
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  • Journal Article

Summary

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Overview

  • This study investigated the presence, diversity, and virulence of Clostridioides difficile (C. difficile), a significant bacterial pathogen, in horses in northwestern France.
  • The research identified toxigenic strains of C. difficile in equidae and examined their potential role in disease and public health from a One Health perspective.

Introduction and Background

  • Clostridioides difficile is an anaerobic, spore-forming bacterium that causes intestinal infections (enteropathogen).
  • While well studied in humans, less is known about its presence and impact in animals, particularly horses (equidae).
  • The One Health concept links human, animal, and environmental health, emphasizing the importance of studying zoonotic pathogens like C. difficile in animals.
  • Horses can live in close contact with humans and may act as reservoirs or vectors for transmission of pathogenic bacteria.

Objectives

  • Evaluate the occurrence and diversity of C. difficile strains in French equidae undergoing necropsy from 2019 to 2021 in northwestern France.
  • Assess the virulence of these strains by detecting toxin genes and free toxins.
  • Determine the potential role of horses in disseminating clinically important C. difficile strains relevant to public health.

Methods

  • Sample Collection:
    • Collected cecal contents and watery intestinal contents systematically from 100 necropsied equidae.
  • Bacterial Isolation:
    • Isolated C. difficile strains anaerobically from intestinal samples.
  • Molecular Characterization:
    • Performed PCR to determine the toxin gene profile of isolated strains.
    • Used PCR ribotyping and compared results with the WEBRIBO database to classify strains.
  • Toxin Detection:
    • Assessed presence of free toxins in intestinal contents to establish clinical relevance.

Key Findings

  • Occurrence: 27 out of 100 equidae tested positive for C. difficile, and 20 had toxigenic strains.
  • Strain Diversity:
    • Toxigenic strains belonged to eight known ribotypes, including ribotypes 078, 126, 005, 012, 020, AI-53, FR227, and notably 017.
    • Non-toxigenic strains were ribotypes 009, 035, and 439.
    • Ribotypes 017 (particularly virulent) and 009 were the most common (five each).
    • Same ribotype strains were found in horses from the same farm, indicating some circulation.
  • Toxin Activity and Disease Correlation:
    • Free toxins (indicating active toxin production) were detected in four horses displaying diarrhea.
    • These horses had either ribotype 126 or ribotype 017 as the sole enteropathogen, suggesting confirmed C. difficile infection (CDI).
    • Three of these animals had prior antibiotic use, a known CDI risk factor.
    • Two horses died from entero-toxic infections caused solely by ribotype 017.
  • Virulence and Public Health Concern:
    • Ribotype 017 showed high virulence and was confirmed as the cause of severe disease and death in some cases.
    • Presence of clinically important ribotypes in horses raises concerns about zoonotic or cross-species transmission.

Conclusions and Implications

  • Approximately 20% of necropsied horses in the study region harbored pathogenic C. difficile strains.
  • The identification of toxigenic and virulent ribotypes—including ribotype 017—in equidae highlights their potential role as reservoirs for pathogens relevant to human and animal health.
  • Horses may contribute to environmental dissemination and cross-species transmission of C. difficile.
  • The findings underscore the importance of surveillance and monitoring of C. difficile in animals under a One Health framework.
  • Routine testing for free toxins in animals with diarrhea may improve diagnosis and understanding of CDI in veterinary medicine.

Importance of the Study

  • Highlights the underexplored role of horses in the ecology of C. difficile.
  • Supports the need for integrated approaches in managing infectious diseases that affect both animals and humans.
  • Raises awareness of antibiotic-associated disease in horses, analogous to hospital-acquired infections in humans.
  • Suggests that controlling C. difficile in animals could reduce risks of zoonotic transmission and environmental contamination.

Cite This Article

APA
Petry S, Tapprest J, Maillard K, Barbut F, Duquesne F, Kozak S, Foucher N, Bernez-Romand M, Bridoux L, Poquet I. (2025). Clostridioides difficile in equidae necropsied in Northwestern France, between 2019 and 2021. Microbiol Spectr, 14(2), e0216525. https://doi.org/10.1128/spectrum.02165-25

Publication

Microbiology spectrum
ISSN: 2165-0497
NlmUniqueID: 101634614
Country: United States
Language: English
Volume: 14
Issue: 2
Pages: e0216525
PII: e02165-25

Researcher Affiliations

Petry, Sandrine
  • ANSES, Normandy Laboratory for Animal Health, Physiopathology and Epidemiology of Equine Diseases Unit (PhEED), Goustranville, Normandy, France.
Tapprest, Jackie
  • ANSES, Normandy Laboratory for Animal Health, Physiopathology and Epidemiology of Equine Diseases Unit (PhEED), Goustranville, Normandy, France.
Maillard, Karine
  • LABÉO Frank Duncombe, Caen, France.
Barbut, Frédéric
  • National Reference Laboratory for C. difficile, Hôpital Saint Antoine, Assistance Publique Hôpitaux de Paris (AP-HP), Paris, France.
  • INSERM S-1139, Université Paris-Cité, Paris, France.
Duquesne, Fabien
  • ANSES, Normandy Laboratory for Animal Health, Physiopathology and Epidemiology of Equine Diseases Unit (PhEED), Goustranville, Normandy, France.
Kozak, Sofia
  • ANSES, Normandy Laboratory for Animal Health, Physiopathology and Epidemiology of Equine Diseases Unit (PhEED), Goustranville, Normandy, France.
Foucher, Nathalie
  • ANSES, Normandy Laboratory for Animal Health, Physiopathology and Epidemiology of Equine Diseases Unit (PhEED), Goustranville, Normandy, France.
Bernez-Romand, Maud
  • ANSES, Normandy Laboratory for Animal Health, Physiopathology and Epidemiology of Equine Diseases Unit (PhEED), Goustranville, Normandy, France.
Bridoux, Ludovic
  • Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, Jouy-en-Josas, France.
Poquet, Isabelle
  • Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, Jouy-en-Josas, France.

MeSH Terms

  • Clostridioides difficile / genetics
  • Clostridioides difficile / isolation & purification
  • Clostridioides difficile / pathogenicity
  • Clostridioides difficile / classification
  • Animals
  • France / epidemiology
  • Clostridium Infections / veterinary
  • Clostridium Infections / microbiology
  • Clostridium Infections / epidemiology
  • Ribotyping
  • Bacterial Toxins / genetics
  • Equidae / microbiology
  • Horse Diseases / microbiology
  • Horse Diseases / epidemiology
  • Horses / microbiology
  • Virulence
  • Autopsy
  • Bacterial Proteins / genetics

Grant Funding

  • 2019-009-Rech CloDifEqui / IFCE
  • 2019-009-Rech CloDifEqui / IFCE
  • SE / Agence Nationale de Sécurité Sanitaire de l'Alimentation, de l'Environnement et du Travail (ANSES)
  • SE / Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement

Conflict of Interest Statement

The authors declare no conflict of interest.

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