FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Sci Rep / The metabolomic profile of a high starch versus no starch di...
Scientific reports2025; 15(1); 35576; doi: 10.1038/s41598-025-23422-z

The metabolomic profile of a high starch versus no starch diet in athletic horses.

Abstract: Feeding a high amount of starch-rich grains is common practice for performance horses even though the horse has evolved to eat a grass based, i.e. low starch diet. To our knowledge, there are no studies using metabolomics to investigate the effects of a high-starch diet in horses. In this study we investigated differences in the plasma metabolic profile of 6 Standardbred horses fed a no-starch, forage-only (F) diet or a high-starch forage-concentrate (FC) diet for 29 days, respectively in a cross-over design. Postprandial plasma samples were collected on the morning of day 25 of each dietary period. Metabolomics analysis of plasma using a targeted H NMR resulted in the quantification of 52 metabolites. Both a univariate and multivariate analysis of metabolites was performed. The univariate analysis found increased (p < 0.05) plasma concentrations of 2-hydroxybutyrate, citrate, dimethyl sulfone, hippurate, methionine, myo-inositol and proline in diet F and higher concentrations of glycine in diet FC. A PLS-DA analysis could discriminate between diets with good predictive power (Q2 (cum) = 0.745, p = 0.032 in CV-ANOVA). We conclude that diet F was strongest identified by metabolites originating from host-microbial co-metabolism and that the clear metabolomic profile discrimination between diets may have implications for health, performance and behaviour. The online version contains supplementary material available at 10.1038/s41598-025-23422-z.
Get Full Text
Publication Date: 2025-10-13 PubMed ID: 41083709PubMed Central: PMC12518770DOI: 10.1038/s41598-025-23422-zGoogle Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

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 research investigated how feeding athletic horses a high-starch diet versus a no-starch, forage-only diet affects their blood plasma metabolite profiles.
  • The study used advanced metabolomics techniques to identify differences in metabolic compounds, which could influence horse health and performance.

Background and Purpose

  • Performance horses are commonly fed starch-rich grains to provide energy, despite their natural evolution towards a grass-based, low-starch diet.
  • There were no previous metabolomics studies assessing how a high-starch diet modifies the metabolic profile in horses.
  • This study aimed to use targeted proton nuclear magnetic resonance (1H NMR) metabolomics to compare the plasma metabolites of horses on two different diets over time.

Study Design

  • Subjects: 6 Standardbred athletic horses.
  • Diets:
    • No-starch, forage-only diet (F).
    • High-starch forage-concentrate diet (FC).
  • Design: Cross-over with each horse fed both diets for 29 days in separate periods.
  • Sample collection: Blood plasma samples collected post-meal on the 25th day of each diet period.

Methods

  • Metabolomics profiling of plasma using targeted 1H NMR spectroscopy.
  • Quantification of 52 different metabolites was achieved.
  • Both univariate (single metabolite differences) and multivariate (pattern analysis) statistical methods applied:
    • Univariate analysis identified specific metabolites that changed significantly.
    • Partial least squares discriminant analysis (PLS-DA) used to classify diets based on metabolite profiles.

Key Findings

  • Univariate analysis results:
    • Higher plasma concentrations of 2-hydroxybutyrate, citrate, dimethyl sulfone, hippurate, methionine, myo-inositol, and proline with the no-starch forage-only diet (F).
    • Higher plasma glycine concentrations with the high-starch forage-concentrate diet (FC).
  • Multivariate PLS-DA analysis:
    • Successfully distinguished between the two diets based on metabolite patterns (Q2 cumulative = 0.745, indicating good predictive power).
    • Significant model validation with CV-ANOVA p = 0.032.
  • Metabolites increased in the forage-only diet suggest greater host-microbial co-metabolism, indicating interactions between the horse’s metabolism and gut microbiota.

Interpretation and Implications

  • The metabolomic differences indicate that diet composition profoundly influences the metabolic processes in athletic horses.
  • The forage-only diet produced a metabolic profile linked to microbial metabolism, which may imply enhanced gut health or different energy utilization compared to the starch-rich diet.
  • The clear discrimination between diets based on plasma metabolites suggests dietary starch level could affect horse health, athletic performance, and possibly behavior.
  • The findings provide a metabolic insight that could guide feeding strategies to optimize health and performance in sport horses.

Additional Information

  • The study’s supplementary materials are available online with detailed data and analysis methods.
  • Published in a peer-reviewed scientific journal (Scientific Reports).

Cite This Article

APA
Nilsson E, Moazzami AA, Lindberg JE, Jansson A. (2025). The metabolomic profile of a high starch versus no starch diet in athletic horses. Sci Rep, 15(1), 35576. https://doi.org/10.1038/s41598-025-23422-z

Publication

Scientific reports
ISSN: 2045-2322
NlmUniqueID: 101563288
Country: England
Language: English
Volume: 15
Issue: 1
Pages: 35576
PII: 35576

Researcher Affiliations

Nilsson, Emma
  • Department of Animal Biosciences, Swedish University of Agricultural Sciences, P.O. Box 7011, 750 07, Uppsala, Sweden. emma.l.nilsson@slu.se.
Moazzami, Ali A
  • Department of Molecular Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Lindberg, Jan Erik
  • Department of Applied Animal Science and Welfare, Swedish University of Agricultural Sciences, P.O. Box 7024, 75007, Uppsala, Sweden.
Jansson, Anna
  • Department of Animal Biosciences, Swedish University of Agricultural Sciences, P.O. Box 7011, 750 07, Uppsala, Sweden.

Conflict of Interest Statement

Declarations. Competing interests: The authors declare no conflict of interest.

References

This article includes 75 references
  1. Jansson A, Harris PA. A bibliometric review on nutrition of the exercising horse from 1970 to 2010.. 169–180 (2013).
    doi: 10.3920/CEP13018google scholar: lookup
  2. Willing B. Changes in faecal bacteria associated with concentrate and forage-only diets fed to horses in training.. 908–914 (2009).
    doi: 10.2746/042516409x447806pubmed: 20383990google scholar: lookup
  3. Hintz HF, Argenzio RA, Schryver HF. Digestion coefficients, blood glucose levels and molar percentage of volatile acids in intestinal fluid of ponies fed varying forage-grain ratios.. 992–995 (1971).
    doi: 10.2527/jas1971.335992xpubmed: 5119977google scholar: lookup
  4. Geor, R. J., Harris, P. & Coenen, M. . (Saunders, 2013).
  5. Bloemen JG. Short chain fatty acids exchange across the gut and liver in humans measured at surgery.. 657–661 (2009).
    doi: 10.1016/j.clnu.2009.05.011pubmed: 19523724google scholar: lookup
  6. Pratt SH, Lawrence LM, Warren LK, Powell DM. The effect of exercise on the clearance of infused acetate in the horse.. 266–271 (2005).
    doi: 10.1016/j.jevs.2005.05.009google scholar: lookup
  7. Emmanuel B. Oxidation of butyrate to ketone bodies and CO2 in the rumen epithelium, liver, kidney, heart and lung of camel (Camelus dromedarius), sheep (Ovis aries) and goat (Carpa hircus).. 699–704 (1980).
    doi: 10.1016/0305-0491(80)90182-0google scholar: lookup
  8. Connysson M, Essén-Gustavsson B, Lindberg JE, Jansson A. Effects of feed deprivation on standardbred horses fed a forage-only diet and a 50:50 forage-oats diet.. 335–340 (2010).
    doi: 10.1111/j.2042-3306.2010.00174.xpubmed: 21059027google scholar: lookup
  9. Connysson M, Muhonen S, Jansson A. Road transport and diet affect metabolic response to exercise in horses.. 4869–4879 (2017).
    doi: 10.2527/jas2017.1670pmc: PMC6292253pubmed: 29293735google scholar: lookup
  10. Jansson A, Lindberg JE. A forage-only diet alters the metabolic response of horses in training.. 1939–1946 (2012).
    doi: 10.1017/s1751731112000948pubmed: 22717208google scholar: lookup
  11. Jansson, A., Connysson, M., Lindberg, J. E. & Dahlborn, K. in (Uppsala Sweden, 2018).
  12. Hothersall B, Nicol C. Role of diet and feeding in normal and stereotypic behaviors in horses.. 167–181 (2009).
    doi: 10.1016/j.cveq.2009.01.002pubmed: 19303558google scholar: lookup
  13. Redondo AJ, Carranza J, Trigo P. Fat diet reduces stress and intensity of startle reaction in horses.. 69–75 (2009).
    doi: 10.1016/j.applanim.2009.02.008google scholar: lookup
  14. Bulmer LS. High-starch diets alter equine faecal microbiota and increase behavioural reactivity.. 18621 (2019).
    doi: 10.1038/s41598-019-54039-8pmc: PMC6901590pubmed: 31819069google scholar: lookup
  15. Valentine BA, Van Saun RJ, Thompson KN, Hintz HF. Role of dietary carbohydrate and fat in horses with equine polysaccharide storage myopathy.. 1537–1544 (2001).
    doi: 10.2460/javma.2001.219.1537pubmed: 11759989google scholar: lookup
  16. Macleay JM, Sorum SA, Valberg SJ, Marsh WE, Sorum MD. Epidemiologic analysis of factors influencing exertional rhabdomyolysis in Thoroughbreds. 1999;60(12):1562–1566.
    doi: 10.2460/ajvr.1999.60.12.1562pubmed: 10622169google scholar: lookup
  17. Luthersson N, Nielsen KH, Harris P, Parkin TDH. Risk factors associated with equine gastric ulceration syndrome (EGUS) in 201 horses in Denmark. 2009;625–630.
    doi: 10.2746/042516409x441929pubmed: 19927579google scholar: lookup
  18. Ametaj BN et al. Metabolomics reveals unhealthy alterations in rumen metabolism with increased proportion of cereal grain in the diet of dairy cows. 2010;583–594.
    doi: 10.1007/s11306-010-0227-6google scholar: lookup
  19. Saleem F et al. A metabolomics approach to uncover the effects of grain diets on rumen health in dairy cows. 2012;6606–6623.
    doi: 10.3168/jds.2012-5403pubmed: 22959937google scholar: lookup
  20. Yang Y et al. Rumen and plasma metabolomics profiling by UHPLC-QTOF/MS revealed metabolic alterations associated with a high-corn diet in beef steers. 2018;e0208031.
    doi: 10.1371/journal.pone.0208031pmc: PMC6261619pubmed: 30485366google scholar: lookup
  21. Goldansaz SA et al. Livestock metabolomics and the livestock metabolome: A systematic review. 2017;e0177675.
    doi: 10.1371/journal.pone.0177675pmc: PMC5439675pubmed: 28531195google scholar: lookup
  22. Bazzano M et al. Exercise induced changes in salivary and serum metabolome in trained standardbred, assessed by (1)H-NMR. 2020;298.
    doi: 10.3390/metabo10070298pmc: PMC7407172pubmed: 32708237google scholar: lookup
  23. Johansson L, Ringmark S, Bergquist J, Skiöldebrand E, Jansson A. A metabolomics perspective on 2 years of high-intensity training in horses. 2024;52188.
    doi: 10.1038/s41598-024-52188-zpmc: PMC10810775pubmed: 38273017google scholar: lookup
  24. Klein DJ, McKeever KH, Mirek ET, Anthony TG. Metabolomic response of equine skeletal muscle to acute fatiguing exercise and training. 2020;110.
    doi: 10.3389/fphys.2020.00110pmc: PMC7040365pubmed: 32132934google scholar: lookup
  25. Leng J et al. Hay versus haylage: Forage type influences the equine urinary metabonome and faecal microbiota. 2022;614–625.
    doi: 10.1111/evj.13456pubmed: 33900659google scholar: lookup
  26. Zhu Y et al. The effect of ryegrass silage feeding on equine fecal microbiota and blood metabolite profile. 2021;715709.
    doi: 10.3389/fmicb.2021.715709pmc: PMC8419423pubmed: 34497595google scholar: lookup
  27. Muhonen S et al. Effects of crude protein intake from grass silage-only diets on the equine colon ecosystem after an abrupt feed change1. 2008;3465–3472.
    doi: 10.2527/jas.2007-0374pubmed: 18676731google scholar: lookup
  28. Muhonen S, Julliand V, Lindberg JE, Bertilsson J, Jansson A. Effects on the equine colon ecosystem of grass silage and haylage diets after an abrupt change from hay1. 2009;2291–2298.
    doi: 10.2527/jas.2008-1461pubmed: 19329474google scholar: lookup
  29. NRC. . (The National Academies Press, 1989).
  30. Lindgren E. The nutritional value of roughages estimated in vivo and by laboratory methods. 61 pp. (1979).
  31. Jansson A. Utfodringsrekommendationer för häst. Department of Animal Nutrition and Management, Uppsala, 2013.
  32. Röhnisch HE et al. AQuA: An automated quantification algorithm for high-throughput nmr-based metabolomics and its application in human plasma. 2095–2102 (2018).
    doi: 10.1021/acs.analchem.7b04324pubmed: 29260864google scholar: lookup
  33. Geor, R. J., Harris, P. A. & Green, E. M.
  34. Lees HJ, Swann JR, Wilson ID, Nicholson JK, Holmes E. Hippurate: The natural history of a mammalian-microbial cometabolite. 1527–1546 (2013).
    doi: 10.1021/pr300900bpubmed: 23342949google scholar: lookup
  35. Besle J-M, Cornu A, Jouany J-P. Roles of structural phenylpropanoids in forage cell wall digestion. 171–190 (1994).
    doi: 10.1002/jsfa.2740640206google scholar: lookup
  36. Rapisarda S, Abu-Ghannam N. Polyphenol characterization and antioxidant capacity of multi-species swards grown in Ireland—Environmental sustainability and nutraceutical potential. 634 (2022).
    doi: 10.3390/su15010634google scholar: lookup
  37. Pallister T et al. Hippurate as a metabolomic marker of gut microbiome diversity: Modulation by diet and relationship to metabolic syndrome. 13670 (2017).
    doi: 10.1038/s41598-017-13722-4pmc: PMC5651863pubmed: 29057986google scholar: lookup
  38. Brial F et al. Human and preclinical studies of the host-gut microbiome co-metabolite hippurate as a marker and mediator of metabolic health. 2105–2114 (2021).
    doi: 10.1136/gutjnl-2020-323314pmc: PMC8515120pubmed: 33975870google scholar: lookup
  39. Escalona EE et al. Dominant components of the Thoroughbred metabolome characterised by (1) H-nuclear magnetic resonance spectroscopy: A metabolite atlas of common biofluids. 721–730 (2015).
    doi: 10.1111/evj.12333pubmed: 25130591google scholar: lookup
  40. Carpio A et al. Evaluation of hippuric acid content in goat milk as a marker of feeding regimen. 5426–5434 (2013).
    doi: 10.3168/jds.2012-6396pubmed: 23849634google scholar: lookup
  41. He X, Slupsky CM. Metabolic fingerprint of dimethyl sulfone (DMSO2) in microbial-mammalian co-metabolism. 5281–5292 (2014).
    doi: 10.1021/pr500629tpubmed: 25245235google scholar: lookup
  42. Butawan M, Benjamin RL, Bloomer RJ. Methylsulfonylmethane: Applications and safety of a novel dietary supplement. 290 (2017).
    doi: 10.3390/nu9030290pmc: PMC5372953pubmed: 28300758google scholar: lookup
  43. Sp N et al. Methylsulfonylmethane inhibits cortisol-induced stress through p53-mediated SDHA/HPRT1 expression in racehorse skeletal muscle cells: A primary step against exercise stress. 214–222 (2020).
    doi: 10.3892/etm.2019.8196pmc: PMC6909739pubmed: 31853292google scholar: lookup
  44. Park JW et al. Regulation of toll-like receptors expression in muscle cells by exercise-induced stress. 1590–1599 (2021).
    doi: 10.5713/ab.20.0484pmc: PMC8495349pubmed: 33332945google scholar: lookup
  45. Yde CC, Bertram HC, Theil PK, Knudsen KEB. Effects of high dietary fibre diets formulated from by-products from vegetable and agricultural industries on plasma metabolites in gestating sows. 460–476 (2011).
    doi: 10.1080/1745039x.2011.621284pubmed: 22256676google scholar: lookup
  46. Owen OE et al. Brain Metabolism during Fasting*. 1589–1595 (1967).
    doi: 10.1172/jci105650pmc: PMC292907pubmed: 6061736google scholar: lookup
  47. Brokner C et al. Metabolic response to dietary fibre composition in horses. 1155–1163 (2016).
    doi: 10.1017/S1751731115003006pubmed: 26755337google scholar: lookup
  48. Sato K et al. Insulin, ketone bodies, and mitochondrial energy transduction. 651–658 (1995).
    doi: 10.1096/fasebj.9.8.7768357pubmed: 7768357google scholar: lookup
  49. Willard JG, Willard JC, Wolfram SA, Baker JP. Effect of diet on cecal pH and feeding behavior of horses. 87–93 (1977).
    doi: 10.2527/jas1977.45187xpubmed: 18431google scholar: lookup
  50. Baldwin RL, Allison MJ. Rumen metabolism. (Suppl 2), 461–477 (1983).
    pubmed: 6352592
  51. Julliand V, de Fombelle A, Drogoul C, Jacotot E. Feeding and microbial disorders in horses: Part 3—Effects of three hay:grain ratios on microbial profile and activities. 543–546 (2001).
    doi: 10.1016/S0737-0806(01)70159-1google scholar: lookup
  52. van Hall G, Sacchetti M, Radegran G. Whole body and leg acetate kinetics at rest, during exercise and recovery in humans. 263–272 (2002).
    doi: 10.1113/jphysiol.2001.014340pmc: PMC2290395pubmed: 12096068google scholar: lookup
  53. Houpt, R. T. in (ed M.J Swenson) 496 (Cornell University Press, Ithaca, New York, USA, 1989).
  54. Essén-Gustavsson B, Connysson M, Jansson A. Effects of crude protein intake from forage-only diets on muscle amino acids and glycogen levels in horses in training. 341–346 (2010).
    doi: 10.1111/j.2042-3306.2010.00283.xpubmed: 21059028google scholar: lookup
  55. Graham-Thiers PM, Bowen LK. Effect of protein source on nitrogen balance and plasma amino acids in exercising horses. 729–735 (2011).
    doi: 10.2527/jas.2010-3081pubmed: 21075968google scholar: lookup
  56. Adeva-Andany M et al. Insulin resistance and glycine metabolism in humans. 11–27 (2018).
    doi: 10.1007/s00726-017-2508-0pubmed: 29094215google scholar: lookup
  57. Alves A, Bassot A, Bulteau AL, Pirola L, Morio B. Glycine metabolism and its alterations in obesity and metabolic diseases. 356 (2019).
    doi: 10.3390/nu11061356pmc: PMC6627940pubmed: 31208147google scholar: lookup
  58. Kreider RB et al. International Society of Sports Nutrition position stand: safety and efficacy of creatine supplementation in exercise, sport, and medicine. 18 (2017).
    doi: 10.1186/s12970-017-0173-zpmc: PMC5469049pubmed: 28615996google scholar: lookup
  59. Schuback K, Essen-Gustavsson B, Persson SG. Effect of creatine supplementation on muscle metabolic response to a maximal treadmill exercise test in Standardbred horses. 533–540 (2000).
    doi: 10.2746/042516400777584578pubmed: 11093628google scholar: lookup
  60. Teixeira FA et al. Oral creatine supplementation on performance of Quarter Horses used in barrel racing. 513–519 (2016).
    doi: 10.1111/jpn.12411pubmed: 26613801google scholar: lookup
  61. Sewell DA, Harris RC. Effects of creatine supplementation in the Thoroughbred horse. 239–242 (1995).
    doi: 10.1111/j.2042-3306.1995.tb04928.xgoogle scholar: lookup
  62. Kohlmeier, M. in (ed Martin Kohlmeier) 265–477 (Academic Press, 2015).
  63. Gall WE et al. alpha-hydroxybutyrate is an early biomarker of insulin resistance and glucose intolerance in a nondiabetic population. e10883 (2010).
    doi: 10.1371/journal.pone.0010883pmc: PMC2878333pubmed: 20526369google scholar: lookup
  64. Wadsworth BJ et al. A 2-hydroxybutyrate-mediated feedback loop regulates muscular fatigue. 92707 (2024).
    doi: 10.7554/eLife.92707pmc: PMC11371357pubmed: 39226092google scholar: lookup
  65. Zhang J et al. Asparagine plays a critical role in regulating cellular adaptation to glutamine depletion. 205–218 (2014).
    doi: 10.1016/j.molcel.2014.08.018pmc: PMC4224619pubmed: 25242145google scholar: lookup
  66. Tanaka H, Takahashi K, Mori M, Ogura M. Metabolism of asparagine and glutamine in growing rats at various dietary protein levels. 1867–1872 (1991).
    doi: 10.1080/00021369.1991.10870873google scholar: lookup
  67. Cederblad G. Effect of diet on plasma carnitine levels and urinary carnitine excretion in humans. 725–729 (1987).
    doi: 10.1093/ajcn/45.4.725pubmed: 3565299google scholar: lookup
  68. Bevilacqua A, Bizzarri M. Inositols in Insulin Signaling and Glucose Metabolism. 1968450 (2018).
    doi: 10.1155/2018/1968450pmc: PMC6286734pubmed: 30595691google scholar: lookup
  69. Raboy V. myo-Inositol-1,2,3,4,5,6-hexakisphosphate. 1033–1043 (2003).
    doi: 10.1016/s0031-9422(03)00446-1pubmed: 14568069google scholar: lookup
  70. Berridge MJ, Irvine RF. Inositol phosphates and cell signalling. 197–205 (1989).
    doi: 10.1038/341197a0pubmed: 2550825google scholar: lookup
  71. Dang NT, Mukai R, Yoshida K-I, Ashida H. D-pinitol and myo-inositol stimulate translocation of glucose transporter 4 in skeletal muscle of C57BL/6 Mice. 1062–1067 (2010).
    doi: 10.1271/bbb.90963pubmed: 20460718google scholar: lookup
  72. Zhang Y et al. Proteomic and metabolomic profiling of a trait anxiety mouse model implicate affected pathways. 008110 (2011).
    doi: 10.1074/mcp.M111.008110pmc: PMC3237072pubmed: 21862759google scholar: lookup
  73. Homer B et al. Microbiota and behavioural issues in production, performance, and companion animals: A systematic review. 458 (2023).
    doi: 10.3390/ani13091458pmc: PMC10177538pubmed: 37174495google scholar: lookup
  74. Palmgren-Karlsson C, Jansson A, Essén-Gustavsson B, Lindberg JE. Effect of molassed sugar beet pulp on nutrient utilisation and metabolic parameters during exercise. 44–49 (2002).
    doi: 10.1111/j.2042-3306.2002.tb05390.xpubmed: 12405658google scholar: lookup
  75. Jansson A et al. Straw as an alternative to grass forage in horses—Effects on post-prandial metabolic profile, energy intake behaviour and gastric ulceration. 2197 (2021).
    doi: 10.3390/ani11082197pmc: PMC8388405pubmed: 34438656google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,005 horse owners like you who receive our email updates on horse health and nutrition.