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Animals : an open access journal from MDPI2025; 15(22); 3343; doi: 10.3390/ani15223343

In Vitro Investigation of Equine Gut Microbiota Alterations During Hypoglycin A Exposure.

Abstract: Hypoglycin A is a plant-derived protoxin that causes atypical myopathy in equids. In atypical myopathy-affected horses, metabolomic and microbiome studies have reported alterations in metabolic markers and faecal microbiota composition, pointing to a potential disruption of microbial homeostasis. However, in vivo observations are strongly confounded by host-related factors, underscoring the need for controlled in vitro approaches. To address this, we used an in vitro static batch fermentation model simulating the equine colon to investigate the direct effects of hypoglycin A on microbiota composition and activity. Faecal inocula from healthy horses were incubated in control and hypoglycin A-treated fermenters for 48 h, with serial analyses of hypoglycin A concentration, short-chain fatty acids, and 16S rRNA gene profiles. Hypoglycin A remained stable in the nutritive medium in the absence of microbiota, confirming that its degradation in inoculated fermenters was microbiota-dependent. The results showed significant microbial-associated hypoglycin A degradation without evidence of toxic metabolite formation. The analysis of α- and β-diversity revealed both an effect of incubation time, reflecting the natural temporal dynamics of microbial communities under batch fermentation, and a specific impact of hypoglycin A exposure, with certain taxa such as being affected. This study provides the first in vitro evidence that the equine microbiota contributes to hypoglycin A degradation.
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Publication Date: 2025-11-19 PubMed ID: 41302050PubMed Central: PMC12649563DOI: 10.3390/ani15223343Google 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.

Equine gut microbiota can degrade hypoglycin A, a plant toxin causing atypical myopathy in horses, as demonstrated in an in vitro colon fermentation model. This study shows hypoglycin A influences microbial community composition and activity without producing toxic metabolites.

Background and Objective

  • Hypoglycin A is a toxin from plants that causes atypical myopathy, a severe muscle disease, in horses.
  • Previous studies in affected horses noted changes in metabolic markers and gut microbiota, suggesting disruption of the microbial balance in the gut.
  • However, studies conducted in living animals (in vivo) are complicated by many host-related factors, making it hard to isolate the effects of hypoglycin A on gut microbes alone.
  • The main objective was to study the direct effects of hypoglycin A on the equine gut microbiota in a controlled laboratory environment (in vitro) using a model simulating the horse colon.

Methods

  • An in vitro static batch fermentation system mimicking the equine colon was used.
  • Faecal samples from healthy horses provided the microbial inoculum to populate the system with natural gut microbes.
  • Two conditions were tested: control fermenters without hypoglycin A and fermenters treated with hypoglycin A, incubated for 48 hours.
  • Regular sampling was performed to analyze:
    • Concentration of hypoglycin A to monitor degradation.
    • Production of short-chain fatty acids (SCFAs), indicators of microbial metabolic activity.
    • 16S rRNA gene sequencing to profile changes in microbial community composition (microbiota diversity).
  • Additional controls included nutrient medium without microbiota to test if hypoglycin A degrades spontaneously.

Key Findings

  • Hypoglycin A remained chemically stable in the nutrient medium alone, confirming it was not degraded abiotically.
  • In the presence of the gut microbiota, significant microbial degradation of hypoglycin A occurred over 48 hours.
  • No toxic metabolites from hypoglycin A were detected, indicating the microbial breakdown did not produce harmful byproducts in this model.
  • Microbial community analysis showed two main effects:
    • Changes associated with fermentation time, reflecting the natural evolution of microbial populations in batch culture.
    • Specific alterations linked to hypoglycin A exposure, including shifts in the abundance of particular microbial taxa (though not all taxa are specified in the abstract).

Implications and Significance

  • This is the first in vitro evidence that the horse gut microbiota can directly degrade hypoglycin A, potentially modulating its toxicity.
  • The absence of toxic metabolites suggests microbial degradation could be a protective mechanism mitigating the toxin’s impact in the gut.
  • Understanding this microbial interaction might help develop strategies to prevent or treat atypical myopathy by targeting or supporting beneficial microbiota.
  • The in vitro model provides a valuable tool to study toxin-microbiota dynamics free from the confounding systemic effects present in living animals.

Cite This Article

APA
François AC, Taminiau B, Renaud B, Gonza-Quito IE, Massey C, Hyde C, Piercy RJ, Douny C, Scippo ML, Daube G, Gustin P, Delcenserie V, Votion DM. (2025). In Vitro Investigation of Equine Gut Microbiota Alterations During Hypoglycin A Exposure. Animals (Basel), 15(22), 3343. https://doi.org/10.3390/ani15223343

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 15
Issue: 22
PII: 3343

Researcher Affiliations

François, Anne-Christine
  • Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Taminiau, Bernard
  • Department of Food Sciences-Microbiology, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Renaud, Benoît
  • Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Gonza-Quito, Irma Elizabeth
  • Laboratory of Food Quality Management, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH)-Veterinary Public Health, University of Liège, 4000 Liège, Belgium.
Massey, Claire
  • Comparative Neuromuscular Diseases Laboratory, Royal Veterinary College, London NW1 0TU, UK.
Hyde, Carolyn
  • Bio-Analysis Centre, London NW1 0NH, UK.
Piercy, Richard J
  • Comparative Neuromuscular Diseases Laboratory, Royal Veterinary College, London NW1 0TU, UK.
Douny, Caroline
  • Laboratory of Food Analysis, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Scippo, Marie-Louise
  • Laboratory of Food Analysis, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Daube, Georges
  • Department of Food Sciences-Microbiology, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Gustin, Pascal
  • Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Delcenserie, Véronique
  • Laboratory of Food Quality Management, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH)-Veterinary Public Health, University of Liège, 4000 Liège, Belgium.
Votion, Dominique-Marie
  • Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.

Grant Funding

  • D31-1381/S1-SAMA / Wallonie agriculture SPW

Conflict of Interest Statement

The authors declare no conflicts of interest.

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