{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:31:23Z","timestamp":1763641883794,"version":"3.45.0"},"reference-count":106,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T00:00:00Z","timestamp":1763510400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Wallonie agriculture SPW","award":["D31-1381\/S1-SAMA"],"award-info":[{"award-number":["D31-1381\/S1-SAMA"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>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 \u03b1- and \u03b2-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 Paraclostridium being affected. This study provides the first in vitro evidence that the equine microbiota contributes to hypoglycin A degradation.<\/jats:p>","DOI":"10.3390\/ani15223343","type":"journal-article","created":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T15:03:15Z","timestamp":1763564595000},"page":"3343","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["In Vitro Investigation of Equine Gut Microbiota Alterations During Hypoglycin A Exposure"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3736-2868","authenticated-orcid":false,"given":"Anne-Christine","family":"Fran\u00e7ois","sequence":"first","affiliation":[{"name":"Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6456-329X","authenticated-orcid":false,"given":"Bernard","family":"Taminiau","sequence":"additional","affiliation":[{"name":"Department of Food Sciences\u2014Microbiology, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9391-3359","authenticated-orcid":false,"given":"Beno\u00eet","family":"Renaud","sequence":"additional","affiliation":[{"name":"Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0076-3473","authenticated-orcid":false,"given":"Irma Elizabeth","family":"Gonza-Quito","sequence":"additional","affiliation":[{"name":"Laboratory of Food Quality Management, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH)\u2014Veterinary Public Health, University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0823-6547","authenticated-orcid":false,"given":"Claire","family":"Massey","sequence":"additional","affiliation":[{"name":"Comparative Neuromuscular Diseases Laboratory, Royal Veterinary College, London NW1 0TU, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolyn","family":"Hyde","sequence":"additional","affiliation":[{"name":"Bio-Analysis Centre, London NW1 0NH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4344-6438","authenticated-orcid":false,"given":"Richard J.","family":"Piercy","sequence":"additional","affiliation":[{"name":"Comparative Neuromuscular Diseases Laboratory, Royal Veterinary College, London NW1 0TU, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4162-1589","authenticated-orcid":false,"given":"Caroline","family":"Douny","sequence":"additional","affiliation":[{"name":"Laboratory of Food Analysis, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8463-3264","authenticated-orcid":false,"given":"Marie-Louise","family":"Scippo","sequence":"additional","affiliation":[{"name":"Laboratory of Food Analysis, Department of Food Sciences, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9282-8408","authenticated-orcid":false,"given":"Georges","family":"Daube","sequence":"additional","affiliation":[{"name":"Department of Food Sciences\u2014Microbiology, Faculty of Veterinary Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pascal","family":"Gustin","sequence":"additional","affiliation":[{"name":"Department of Functional Sciences, Faculty of Veterinary Medicine, Pharmacology and Toxicology, Fundamental and Applied Research for Animals & Health (FARAH), University of Li\u00e8ge, 4000 Li\u00e8ge, 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Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1111\/evj.12117","article-title":"Identification of Methylenecyclopropyl Acetic Acid in Serum of European Horses with Atypical Myopathy","volume":"46","author":"Votion","year":"2014","journal-title":"Equine Vet. J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1111\/evj.12499","article-title":"Samaras and Seedlings of Acer Pseudoplatanus Are Potential Sources of Hypoglycin A Intoxication in Atypical Myopathy without Necessarily Inducing Clinical Signs","volume":"48","author":"Baise","year":"2016","journal-title":"Equine Vet. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1111\/jvim.12367","article-title":"Hypoglycin A Concentrations in Seeds of Acer Pseudoplatanus Trees Growing on Atypical Myopathy-Affected and Control Pastures","volume":"28","author":"Unger","year":"2014","journal-title":"J. Vet. Intern. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1016\/0031-9422(73)80387-5","article-title":"Cyclopropylamino Acids of the Genus Acer","volume":"12","author":"Fowden","year":"1973","journal-title":"Phytochemistry"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bochnia, M., Sander, J., Ziegler, J., Terhardt, M., Sander, S., Janzen, N., Cavalleri, J.M.V., Zuraw, A., Wensch-Dorendorf, M., and Zeyner, A. (2019). Detection of MCPG Metabolites in Horses with Atypical Myopathy. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0211698"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.1016\/S0040-4039(01)90775-0","article-title":"A New Synthesis of Hypoglycin a","volume":"4","author":"Black","year":"1963","journal-title":"Tetrahedron Lett."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1016\/S0076-6879(81)72051-2","article-title":"Hypoglycin and Metabolically Related Inhibitors","volume":"72","author":"Billington","year":"1981","journal-title":"Methods Enzymol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"El-Khatib, A.H., Engel, A.M., and Weigel, S. (2022). Co-Occurrence of Hypoglycin A and Hypoglycin B in Sycamore and Box Elder Maple Proved by LC-MS\/MS and LC-HR-MS. Toxins, 14.","DOI":"10.3390\/toxins14090608"},{"key":"ref_9","first-page":"125","article-title":"Studies of the Action of Hypoglycin-A, an Hypoglycaemic Substance","volume":"13","author":"Feng","year":"1958","journal-title":"J. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3537","DOI":"10.1093\/jas\/sky200","article-title":"Atypical Myopathy in P\u00e8re David\u2019s Deer (Elaphurus davidianus) Associated with Ingestion of Hypoglycin A","volume":"96","author":"Bunert","year":"2018","journal-title":"J. Anim. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Renaud, B., Kruse, C.J., Fran\u00e7ois, A.C., Grund, L., Bunert, C., Brisson, L., Boemer, F., Gault, G., Ghislain, B., and Petitjean, T. (2022). Acer Pseudoplatanus: A Potential Risk of Poisoning for Several Herbivore Species. Toxins, 14.","DOI":"10.3390\/toxins14080512"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1016\/S0022-3565(25)11947-2","article-title":"Pharmacologic Action of Hypoglycin A and B","volume":"121","author":"Chen","year":"1957","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1038\/173356b0","article-title":"Hypoglycin A, B: Biologically Active Polypeptides from Blighia sapida","volume":"173","author":"Hassall","year":"1954","journal-title":"Nature"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1177\/10406387211020721","article-title":"Atypical Myopathy in 2 Bactrian Camels","volume":"33","author":"Hirz","year":"2021","journal-title":"J. Vet. Diagn. Investig."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1093\/oxfordjournals.aje.a118321","article-title":"The Vomiting Sickness of Jamaica, BWI and Its Relation to Akee Poisoning","volume":"25","author":"Jordan","year":"1937","journal-title":"Am. J. Epidemiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1056\/NEJM197608262950901","article-title":"Jamaican Vomiting Sickness: Biochemical Investigation of Two Cases","volume":"295","author":"Tanaka","year":"1976","journal-title":"N. Engl. J. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"104515","DOI":"10.1016\/j.etap.2024.104515","article-title":"Large-Scale Study of Blood Markers in Equine Atypical Myopathy Reveals Subclinical Poisoning and Advances in Diagnostic and Prognostic Criteria","volume":"110","author":"Renaud","year":"2024","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kruse, C.-J., Stern, D., Mouithys-Mickalad, A., Niesten, A., Art, T., Lemieux, H., and Votion, D.-M. (2021). In Vitro Assays for the Assessment of Impaired Mitochondrial Bioenergetics in Equine Atypical Myopathy. Life, 11.","DOI":"10.3390\/life11070719"},{"key":"ref_19","first-page":"1","article-title":"Methylenecyclopropaneacetic Acid, a Metabolite of Hypoglycin","volume":"3","year":"1966","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Boemer, F., Detilleux, J., Cello, C., Amory, H., Marcillaud-Pitel, C., Richard, E., Van Galen, G., Van Loon, G., Lef\u00e8re, L., and Votion, D.M. (2017). Acylcarnitines Profile Best Predicts Survival in Horses with Atypical Myopathy. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0182761"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.tvjl.2016.07.015","article-title":"Equine Atypical Myopathy: A Metabolic Study","volume":"216","author":"Jahn","year":"2016","journal-title":"Vet. J."},{"key":"ref_22","first-page":"100827","article-title":"Diagnosis of Atypical Myopathy Based on Organic Acid and Acylcarnitine Profiles and Evolution of Biomarkers in Surviving Horses","volume":"29","author":"Mathis","year":"2021","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Bochnia, M., Ziegler, J., Sander, J., Uhlig, A., Schaefer, S., Vollstedt, S., Glatter, M., Abel, S., Recknagel, S., and Schusser, G.F. (2015). Hypoglycin a Content in Blood and Urine Discriminates Horses with Atypical Myopathy from Clinically Normal Horses Grazing on the Same Pasture. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0136785"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1016\/j.nmd.2008.02.007","article-title":"Acquired Multiple Acyl-CoA Dehydrogenase Deficiency in 10 Horses with Atypical Myopathy","volume":"18","author":"Westermann","year":"2008","journal-title":"Neuromuscul. Disord."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1016\/j.ymgme.2007.04.010","article-title":"Equine Biochemical Multiple Acyl-CoA Dehydrogenase Deficiency (MADD) as a Cause of Rhabdomyolysis","volume":"91","author":"Westermann","year":"2007","journal-title":"Mol. Genet. Metab."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1967","DOI":"10.1016\/j.cell.2025.01.045","article-title":"Functional Diversification of Dietary Plant Small Molecules by the Gut Microbiome","volume":"188","author":"Kuziel","year":"2025","journal-title":"Cell"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Tan, Y., An, K., and Su, J. (2024). Review: Mechanism of Herbivores Synergistically Metabolizing Toxic Plants through Liver and Intestinal Microbiota. Comp. Biochem. Physiol. Part C Toxicol. Pharmacol., 281.","DOI":"10.1016\/j.cbpc.2024.109925"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1111\/1462-2920.16507","article-title":"Microbial Degradation of Plant Toxins","volume":"25","author":"Coolen","year":"2023","journal-title":"Environ. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Engel, A.M., El-Khatib, A.H., Bachmann, M., Wensch-Dorendorf, M., Klevenhusen, F., Weigel, S., Pieper, R., and Zeyner, A. (2025). Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures. Toxins, 17.","DOI":"10.3390\/toxins17020046"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.tvjl.2019.05.005","article-title":"Methods and Basic Concepts for Microbiota Assessment","volume":"249","author":"Costa","year":"2019","journal-title":"Vet. J."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Wimmer-Scherr, C., Taminiau, B., Renaud, B., van Loon, G., Palmers, K., Votion, D., Amory, H., Daube, G., and Cesarini, C. (2021). Comparison of Fecal Microbiota of Horses Suffering from Atypical Myopathy and Healthy Co-Grazers. Animals, 11.","DOI":"10.3390\/ani11020506"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Fran\u00e7ois, A.-C., Cesarini, C., Taminiau, B., Renaud, B., Kruse, C.-J., Boemer, F., van Loon, G., Palmers, K., Daube, G., and Wouters, C.P. (2025). Unravelling Faecal Microbiota Variations in Equine Atypical Myopathy: Correlation with Blood Markers and Contribution of Microbiome. Animals, 15.","DOI":"10.3390\/ani15030354"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"102943","DOI":"10.1016\/j.jevs.2020.102943","article-title":"Factors Influencing Equine Gut Microbiota: Current Knowledge","volume":"88","author":"Garber","year":"2020","journal-title":"J. Equine Vet. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Boucher, L., Leduc, L., Lecl\u00e8re, M., and Costa, M.C. (2024). Current Understanding of Equine Gut Dysbiosis and Microbiota Manipulation Techniques: Comparison with Current Knowledge in Other Species. Animals, 14.","DOI":"10.3390\/ani14050758"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e02093-19","DOI":"10.1128\/AEM.02093-19","article-title":"\u201cBowel on the Bench\u201d: Proof of Concept of a Three-Stage, In Vitro Fermentation Model of the Equine Large Intestine","volume":"86","author":"Leng","year":"2020","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/S0377-8401(99)00039-5","article-title":"Evulation of an In Vitro Batch Culture Technique for Estimating the In Vivo Digestibility and Digestible Energy Content of Equine Feeds Using Equine Faeces as the Source of Microbial Inoculum","volume":"80","author":"Lowman","year":"1999","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.jchromb.2015.06.004","article-title":"Quantification of Hypoglycin A in Serum Using ATRAQ\u00ae Assay","volume":"997","author":"Boemer","year":"2015","journal-title":"J. Chromatogr. B"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1111\/j.2042-3306.2012.00684.x","article-title":"Seasonal Pasture Myopathy\/Atypical Myopathy in North America Associated with Ingestion of Hypoglycin A within Seeds of the Box Elder Tree","volume":"45","author":"Valberg","year":"2013","journal-title":"Equine Vet. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mimet.2014.08.022","article-title":"To Pool or Not to Pool? Impact of the Use of Individual and Pooled Fecal Samples for In Vitro Fermentation Studies","volume":"107","author":"Aguirre","year":"2014","journal-title":"J. Microbiol. Methods"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.fct.2005.07.002","article-title":"Ackee (Blighia Sapida) Hypoglycin A Toxicity: Dose Response Assessment in Laboratory Rats","volume":"44","author":"Blake","year":"2006","journal-title":"Food Chem. Toxicol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"27","DOI":"10.4103\/0976-0105.177703","article-title":"A Simple Practice Guide for Dose Conversion between Animals and Human","volume":"7","author":"Nair","year":"2016","journal-title":"J. Basic Clin. Pharm."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.jevs.2019.02.013","article-title":"The Influence of Body Mass and Height on Equine Hoof Conformation and Symmetry","volume":"77","author":"Whittington","year":"2019","journal-title":"J. Equine Vet. Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1097\/01.NURSE.0000558092.00476.db","article-title":"Take a Deeper Look into Body Surface Area","volume":"49","author":"Smith","year":"2019","journal-title":"Nursing"},{"key":"ref_44","first-page":"13","article-title":"Digestive Physiology of the Horse","volume":"46","author":"Hintz","year":"1975","journal-title":"J. S. Afr. Vet. Assoc."},{"key":"ref_45","first-page":"191","article-title":"Validation of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) Reactor Using Microorganism-Associated Activities","volume":"7","author":"Molly","year":"1994","journal-title":"Microb. Ecol. Health Dis."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"113348","DOI":"10.1016\/j.foodres.2023.113348","article-title":"Butyrogenic, Bifidogenic and Slight Anti-Inflammatory Effects of a Green Kiwifruit Powder (Kiwi FFG\u00ae) in a Human Gastrointestinal Model Simulating Mild Constipation","volume":"173","author":"Bondue","year":"2023","journal-title":"Food Res. Int."},{"key":"ref_47","unstructured":"Geor, R.J., Harris, P.A., and Coenen, M. (2013). Gastrointestinal Physiology. Equine Applied and Clinical Nutrition, W.B. Saunders."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1080\/15563650600584410","article-title":"Ackee Fruit Poisoning: An Outbreak Investigation in Haiti 2000-2001, and Review of the Literature","volume":"44","author":"Joskow","year":"2006","journal-title":"Clin. Toxicol."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Wang, Y., and Qian, P.Y. (2009). Conservative Fragments in Bacterial 16S RRNA Genes and Primer Design for 16S Ribosomal DNA Amplicons in Metagenomic Studies. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0007401"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2727","DOI":"10.1111\/jvim.15937","article-title":"Effect of Oral Administration of Omeprazole on the Microbiota of the Gastric Glandular Mucosa and Feces of Healthy Horses","volume":"34","author":"Cerri","year":"2020","journal-title":"J. Vet. Intern. Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"7537","DOI":"10.1128\/AEM.01541-09","article-title":"Introducing Mothur: Open-Source, Platform-Independent, Community-Supported Software for Describing and Comparing Microbial Communities","volume":"75","author":"Schloss","year":"2009","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e2584","DOI":"10.7717\/peerj.2584","article-title":"VSEARCH: A Versatile Open Source Tool for Metagenomics","volume":"2016","author":"Rognes","year":"2016","journal-title":"PeerJ"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"D590","DOI":"10.1093\/nar\/gks1219","article-title":"The SILVA Ribosomal RNA Gene Database Project: Improved Data Processing and Web-Based Tools","volume":"41","author":"Quast","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_54","unstructured":"Oksanen, J., Simpso, G.L., Blanchet, F.G., Kindt, R., Legendre, P., Minchin, P.R., O\u2019Hara, R.B., Solymos, P., Stevens, M.H.M., and Szoecs, E. (2024, October 09). Package \u201cvegan\u201d\u2014Community Ecology Package. Available online: https:\/\/cran.r-project.org\/web\/packages\/vegan\/vegan.pdf."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Kers, J.G., and Saccenti, E. (2022). The Power of Microbiome Studies: Some Considerations on Which Alpha and Beta Metrics to Use and How to Report Results. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.796025"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"3514","DOI":"10.1002\/ece3.1155","article-title":"Choosing and Using Diversity Indices: Insights for Ecological Applications from the German Biodiversity Exploratories","volume":"4","author":"Morris","year":"2014","journal-title":"Ecol. Evol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1111\/j.1461-0248.2004.00707.x","article-title":"A New Statistical Approach for Assessing Similarity of Species Composition with Incidence and Abundance Data","volume":"8","author":"Chao","year":"2005","journal-title":"Ecol. Lett."},{"key":"ref_58","unstructured":"Oksanen, J., Kindt, R., and Simpson, G. (2024, October 09). Package \u201cvegan3d\u201d\u2014Static and Dynamic 3D and Editable Interactive Plots for the \u201cVegan\u201d Package. Available online: https:\/\/cran.r-project.org\/web\/packages\/vegan3d\/vegan3d.pdf."},{"key":"ref_59","unstructured":"Martinez Arbizu, P. (2024, October 09). PairwiseAdonis: Pairwise Multilevel Comparison Using Adonis_. R Package Version 0.4.1. Available online: https:\/\/github.com\/pmartinezarbizu\/pairwiseAdonis."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/j.jchromb.2019.06.013","article-title":"Development of an Analytical Method to Detect Short-Chain Fatty Acids by SPME-GC\u2013MS in Samples Coming from an In Vitro Gastrointestinal Model","volume":"1124","author":"Douny","year":"2019","journal-title":"J. Chromatogr. B"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1111\/evj.13303","article-title":"Detection of Hypoglycin A and MCPA-Carnitine in Equine Serum and Muscle Tissue: Optimisation and Validation of a LC-MS-Based Method without Derivatisation","volume":"53","author":"Hyde","year":"2021","journal-title":"Equine Vet. J."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"102600","DOI":"10.1016\/j.anaerobe.2022.102600","article-title":"Reconsidering the In Vivo Functions of Clostridial Stickland Amino Acid Fermentations","volume":"76","author":"Pavao","year":"2022","journal-title":"Anaerobe"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1139\/m82-043","article-title":"Leucine Dissimilation to Isovaleric and Isocaproic Acids by Cell Suspensions of Amino Acid Fermenting Anaerobes: The Stickland Reaction Revisited","volume":"28","author":"Britz","year":"1981","journal-title":"Can. J. Microbiol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1768","DOI":"10.2741\/3820","article-title":"Amino Acid Metabolism in Intestinal Bacteria-Links between Gut Ecology and Host Health","volume":"16","author":"Dai","year":"2011","journal-title":"Front. Biosci."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.phrs.2012.11.005","article-title":"Intestinal Luminal Nitrogen Metabolism: Role of the Gut Microbiota and Consequences for the Host","volume":"68","author":"Davila","year":"2013","journal-title":"Pharmacol. Res."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Granado-Serrano, A.B., Mart\u00edn-Gar\u00ed, M., S\u00e1nchez, V., Riart Solans, M., Berd\u00fan, R., Ludwig, I.A., Rubi\u00f3, L., Vilapriny\u00f3, E., Portero-Ot\u00edn, M., and Serrano, J.C.E. (2019). Faecal Bacterial and Short-Chain Fatty Acids Signature in Hypercholesterolemia. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-38874-3"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1152\/physrev.1990.70.2.567","article-title":"Energy Contributions of Volatile Fatty Acids from the Gastrointestinal Tract in Various Species","volume":"70","author":"Bergman","year":"1990","journal-title":"Physiol. Rev."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1111\/j.1574-6968.2002.tb11467.x","article-title":"The Microbiology of Butyrate Formation in the Human Colon","volume":"217","author":"Pryde","year":"2002","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1079\/BJN20041150","article-title":"Contribution of Acetate to Butyrate Formation by Human Faecal Bacteria","volume":"91","author":"Duncan","year":"2004","journal-title":"Br. J. Nutr."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1574-6968.2009.01514.x","article-title":"Diversity, Metabolism and Microbial Ecology of Butyrate-Producing Bacteria from the Human Large Intestine","volume":"294","author":"Louis","year":"2009","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1170","DOI":"10.1111\/jvim.16077","article-title":"Hypoglycin A Absorption in Sheep without Concurrent Clinical or Biochemical Evidence of Disease","volume":"35","author":"Bevin","year":"2021","journal-title":"J. Vet. Intern. Med."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"809","DOI":"10.1002\/anie.197908093","article-title":"Bonding Properties of Cyclopropane and Their Chemical Consequences","volume":"18","year":"1979","journal-title":"Angew. Chem."},{"key":"ref_73","unstructured":"Freeman, W.H. (2021). Principles of Biochemistry, Macmillan International. [8th ed.]."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/S0031-6997(25)06873-5","article-title":"Hypoglycin and Hypoglycin-like Compounds","volume":"21","author":"Bressler","year":"1969","journal-title":"Pharmacol. Rev."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1101\/SQB.1961.026.01.031","article-title":"Catabolite Repression","volume":"26","author":"Magasanik","year":"1961","journal-title":"Cold Spring Harb. Symp. Quant. Biol."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"1803","DOI":"10.1021\/ja01199a067","article-title":"The Structure of Leucenol","volume":"69","author":"Adams","year":"1947","journal-title":"J. Am. Chem. Soc."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"1801","DOI":"10.1021\/ja01199a066","article-title":"On the Structure of Leucaenine (Leucaenol) from Leucaena Glauca Bentham","volume":"69","author":"Bickel","year":"1947","journal-title":"J. Am. Chem. Soc."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1111\/j.1751-0813.1981.tb07097.x","article-title":"Does Ruminal Metabolism of Mimosine Explain the Absence of Leucaena Toxicity in Hawaii?","volume":"57","author":"Jones","year":"1981","journal-title":"Aust. Vet. J."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1435","DOI":"10.1007\/BF01951931","article-title":"Australian Goats Detoxify the Goitrogen 3-Hydroxy-4(IH) Pyridone (DHP) after Rumen Infusion from an Indone-Sian Goat","volume":"40","author":"Jones","year":"1984","journal-title":"Experientia"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1111\/j.1751-0813.1986.tb02990.x","article-title":"Successful Transfer of DHP-Degrading Bacteria from Hawaiian Goats to Australian Ruminants to Overcome the Toxicity of Leucaena","volume":"63","author":"Jones","year":"1986","journal-title":"Aust. Vet. J."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1016\/S0723-2020(11)80111-6","article-title":"Synergistes jonesii, gen. nov., sp.nov.: A Rumen Bacterium That Degrades Toxic Pyridinediols","volume":"15","author":"Allison","year":"1992","journal-title":"Syst. Appl. Microbiol."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1128\/aem.56.3.590-594.1990","article-title":"Detection of Ruminal Bacteria That Degrade Toxic Dihydroxypyridine Compounds Produced from Mimosine","volume":"56","author":"Allison","year":"1990","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_83","doi-asserted-by":"crossref","unstructured":"Baraille, M., Buttet, M., Grimm, P., Milojevic, V., Julliand, S., and Julliand, V. (2024). Changes of Faecal Bacterial Communities and Microbial Fibrolytic Activity in Horses Aged from 6 to 30 Years Old. PLoS ONE, 19.","DOI":"10.1371\/journal.pone.0303029"},{"key":"ref_84","doi-asserted-by":"crossref","unstructured":"Theelen, M.J.P., Luiken, R.E.C., Wagenaar, J.A., van Oldruitenborgh-Oosterbaan, M.M.S., Rossen, J.W.A., and Zomer, A.L. (2021). The Equine Faecal Microbiota of Healthy Horses and Ponies in The Netherlands: Impact of Host and Environmental Factors. Animals, 11.","DOI":"10.3390\/ani11061762"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"103355","DOI":"10.1016\/j.jevs.2020.103355","article-title":"Effect of Gender Bias on Equine Fecal Microbiota","volume":"97","author":"Hu","year":"2021","journal-title":"J. Equine Vet. Sci."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1111\/j.2042-3306.2012.00555.x","article-title":"European Outbreaks of Atypical Myopathy in Grazing Horses (2006\u20132009): Determination of Indicators for Risk and Prognostic Factors","volume":"44","author":"Saegerman","year":"2012","journal-title":"Equine Vet. J."},{"key":"ref_87","doi-asserted-by":"crossref","unstructured":"Votion, D.M., Fran\u00e7ois, A.C., Kruse, C., Renaud, B., Farinelle, A., Bouquieaux, M.C., Marcillaud-pitel, C., and Gustin, P. (2020). Answers to the Frequently Asked Questions Regarding Horse Feeding and Management Practices to Reduce the Risk of Atypical Myopathy. Animals, 10.","DOI":"10.3390\/ani10020365"},{"key":"ref_88","doi-asserted-by":"crossref","unstructured":"Fernandes, K.A., Rogers, C.W., Gee, E.K., Kittelmann, S., Bolwell, C.F., Bermingham, E.N., Biggs, P.J., and Thomas, D.G. (2021). Resilience of Faecal Microbiota in Stabled Thoroughbred Horses Following Abrupt Dietary Transition between Freshly Cut Pasture and Three Forage-Based Diets. Animals, 11.","DOI":"10.3390\/ani11092611"},{"key":"ref_89","doi-asserted-by":"crossref","unstructured":"Fernandes, K.A., Gee, E.K., Rogers, C.W., Kittelmann, S., Biggs, P.J., Bermingham, E.N., Bolwell, C.F., and Thomas, D.G. (2021). Seasonal Variation in the Faecal Microbiota of Mature Adult Horses Maintained on Pasture in New Zealand. Animals, 11.","DOI":"10.3390\/ani11082300"},{"key":"ref_90","doi-asserted-by":"crossref","unstructured":"Raspa, F., Chessa, S., Bergero, D., Sacchi, P., Ferrocino, I., Cocolin, L., Corvaglia, M.R., Moretti, R., Cavallini, D., and Valle, E. (2024). Microbiota Characterization throughout the Digestive Tract of Horses Fed a High-Fiber vs. a High-Starch Diet. Front. Vet. Sci., 11.","DOI":"10.3389\/fvets.2024.1386135"},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"Salem, S.E., Maddox, T.W., Berg, A., Antczak, P., Ketley, J.M., Williams, N.J., and Archer, D.C. (2018). Variation in Faecal Microbiota in a Group of Horses Managed at Pasture over a 12-Month Period. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-26930-3"},{"key":"ref_92","doi-asserted-by":"crossref","unstructured":"Chaucheyras-Durand, F., Sacy, A., Karges, K., and Apper, E. (2022). Gastro-Intestinal Microbiota in Equines and Its Role in Health and Disease: The Black Box Opens. Microorganisms, 10.","DOI":"10.3390\/microorganisms10122517"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"e18860","DOI":"10.7717\/peerj.18860","article-title":"Changes in Bacterial Viability after Preparation and Storage of Fecal Microbiota Transplantation Solution Using Equine Feces","volume":"13","author":"Arantes","year":"2025","journal-title":"PeerJ"},{"key":"ref_94","doi-asserted-by":"crossref","unstructured":"Fouhy, F., Deane, J., Rea, M.C., O\u2019Sullivan, \u00d3., Ross, R.P., O\u2019Callaghan, G., Plant, B.J., and Stanton, C. (2015). The Effects of Freezing on Faecal Microbiota as Determined Using Miseq Sequencing and Culture-Based Investigations. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0119355"},{"key":"ref_95","doi-asserted-by":"crossref","unstructured":"Carroll, I.M., Ringel-Kulka, T., Siddle, J.P., Klaenhammer, T.R., and Ringel, Y. (2012). Characterization of the Fecal Microbiota Using High-Throughput Sequencing Reveals a Stable Microbial Community during Storage. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0046953"},{"key":"ref_96","doi-asserted-by":"crossref","unstructured":"De Bustamante, M.M., Plummer, C., Macnicol, J., and Gomez, D. (2021). Impact of Ambient Temperature Sample Storage on the Equine Fecal Microbiota. Animals, 11.","DOI":"10.3390\/ani11030819"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"106220","DOI":"10.1016\/j.tvjl.2024.106220","article-title":"Storage of Equine Faecal Microbiota Transplantation Solution Has Minimal Impact on Major Bacterial Communities and Structure","volume":"307","author":"Bell","year":"2024","journal-title":"Vet. J."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"e10837","DOI":"10.7717\/peerj.10837","article-title":"Long-Term Storage of Feces at \u221280 \u00b0C versus \u221220 \u00b0C Is Negligible for 16S RRNA Amplicon Profiling of the Equine Bacterial Microbiome","volume":"9","author":"Gavriliuc","year":"2021","journal-title":"PeerJ"},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1111\/evj.12723","article-title":"Identification of Hypoglycin A Binding Adsorbents as Potential Preventive Measures in Co-Grazers of Atypical Myopathy Affected Horses","volume":"50","author":"Cavalleri","year":"2018","journal-title":"Equine Vet. J."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"679","DOI":"10.5713\/ajas.20.0050","article-title":"Amino Acid Requirements in Horses","volume":"33","author":"Mok","year":"2020","journal-title":"Asian-Australas. J. Anim. Sci."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"400","DOI":"10.2527\/jas1988.662400x","article-title":"Digestion of Hay Protein in Different Segments of the Equine Digestive Tract","volume":"66","author":"Gibbs","year":"1988","journal-title":"J. Anim. Sci."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"1190","DOI":"10.2527\/jas1972.3561190x","article-title":"Digestion and Utilization of Cecally Infused Protein by the Equine","volume":"35","author":"Reitnour","year":"1972","journal-title":"J. Anim. Sci."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"943","DOI":"10.2527\/jas1983.574943x","article-title":"Nitrogen Partitioning Along the Equine Digestive Tract","volume":"57","author":"Glade","year":"1983","journal-title":"J. Anim. Sci."},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"1028","DOI":"10.2527\/jas.2009-2406","article-title":"Cationic and Neutral Amino Acid Transporter Transcript Abundances Are Differentially Expressed in the Equine Intestinal Tract","volume":"88","author":"Woodward","year":"2010","journal-title":"J. Anim. Sci."},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/S0007-1935(17)37583-8","article-title":"Digestion and Absorption of 15N-Labelled Microbial Protein in the Large Intestine of the Horse","volume":"127","author":"Slade","year":"1971","journal-title":"Br. Vet. J."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"2930","DOI":"10.3390\/nu7042930","article-title":"The Role of Microbial Amino Acid Metabolism in Host Metabolism","volume":"7","author":"Neis","year":"2015","journal-title":"Nutrients"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/22\/3343\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:19:25Z","timestamp":1763641165000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/22\/3343"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,19]]},"references-count":106,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ani15223343"],"URL":"https:\/\/doi.org\/10.3390\/ani15223343","relation":{},"ISSN":["2076-2615"],"issn-type":[{"type":"electronic","value":"2076-2615"}],"subject":[],"published":{"date-parts":[[2025,11,19]]}}}