Oxidative stress biomarkers and free amino acid concentrations in the blood plasma of moderately exercised horses indicate adaptive response to prolonged exercise training.
Abstract: Oxidative stress caused by routine physical stressors may negatively impact the performance of equine athletes; thus, the present study identifies oxidative biomarkers in the blood plasma of exercising horses. Stock-type horses were subject to a standardized moderate-intensity exercise protocol 3 times per week for 8 wk. Exercise protocol followed NRC guidelines consisting of 30% walk, 55% trot, and 15% canter, with a target heart rate (HR) of 90 BPM. Blood plasma was collected in wk 1, 2, 7, and 8 immediately before and 0, 30, 60, and 90 min after exercise and analyzed for total antioxidant capacity (TAC), thiobarbituric acid reactive substance (TBARS), glutathione peroxidase activity (GPx), and superoxide dismutase activity (SOD). Data were analyzed as repeated measures with wk, d, time, and their interactions as fixed effects. The TAC on day 2 (0.40 mM Trolox) was 7.5% greater than on day 3 (P = 0.013). There were wk × d × time interactions for SOD, TBARS, and GPx (P < 0.001). The TBARS remained at pre-exercise baseline (d-1 wk-1; 2.7 µM malondialdehyde) for most collection times within weeks 1, 7, and 8 (P ≥ 0.058); however, TBARS increased by 0.24 to 0.41 µM on day 2 of week 2 post-exercise (P < 0.001) and remained similarly elevated on day 3 pre- and immediately post-exercise (P < 0.001). The GPx similarly remained at baseline (172.6 µM/min; P ≥ 0.621) but increased by 48.18 to 83.4 µM/min at most collection times on days 1 and 2 of week 2 (P ≤ 0.023). The SOD remained at baseline (167.2 U/ mL; P ≥ 0.055) until increasing by 11.28 to 15.61 U/mL at 30 min post-exercise on day 1, week 1 and at most collection times on day 3, week 8 (P ≤ 0.043). Amino acids with antioxidant properties such as Met, Tyr, and Trp drastically decreased from weeks 2 to 8 (P < 0.001). Met and Tyr also decreased from -60 to 90 min (P < 0.047), whereas there was no time effect on Trp concentration (P = 0.841). The current study indicates the time-dependent nature of oxidative stress concerning persistent stressors such as exercise. Performance horses are subjected to numerous stressors. These stressors may subsequently impact their overall performance. The present study measured oxidative stress biomarkers in the blood of exercising horses. Horses were moderately exercised over an 8-wk period and blood plasma was collected to measure total antioxidant capacity (TAC), thiobarbituric acid reactive substance (TBARS), glutathione peroxidase activity (GPx), and superoxide dismutase activity (SOD). Amino acid concentration was also evaluated. The TAC was greater on day 2 vs. day 3. The TBARS remained at pre-exercise (baseline) at most times except for increasing on day 2 of week 2 post-exercise. The GPx also remained at baseline for most times but increased on days 1 and 2 of week 2. The SOD remained at baseline until increasing at 30 min post-exercise on day 1, week 1 and at most collection times on day 3, week 8. Amino acids with antioxidant properties drastically decreased from weeks 2 to 8. Horses are exposed to a variety of physical stressors on a regular basis that may produce similar effects in the equine stress response. Understanding the response in the equine athlete when exposed to new stressors is crucial in determining how to prevent oxidative damage in future athletes.
© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science.
All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
Publication Date: 2022-03-18 PubMed ID: 35298640PubMed Central: PMC9030216DOI: 10.1093/jas/skac086Google 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.
- 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.
This research explores how stress markers and amino acids found in horse’s blood plasma adjust in response to moderate, prolonged exercise. The study finds out that performance horses demonstrate a time-dependent adjustment to exercise stress, which is reflected in specific oxidative stress biomarkers and a decrease in certain amino acids.
Study Setup and Methods
- The research focused on stock-type horses which underwent a standard moderate-intensity exercise protocol three times a week over an 8-week period. The designed exercise pattern followed Nutrition Requirements of Horses guidelines which contained walk (30%), trot (55%), and canter (15%) at a target heart rate of 90 BPM.
- Blood plasma was collected on specific days and times (immediately before and 0, 30, 60, 90 minutes after exercise). This was done in the first, second, seventh, and eighth week.
- The blood plasma was then examined for several indicators. These were the total antioxidant capacity (TAC), thiobarbituric acid reactive substance (TBARS), glutathione peroxidase activity (GPx), and superoxide dismutase activity (SOD). Any interactions and fixed effects influenced by day, duration, and week were statistically analyzed.
Findings
- The research found certain variations in the recorded measurements. For instance, the total antioxidant capacity (TAC) on the second day showed a 7.5% increase compared to the third day.
- However, by observing the mark changes in the oxidative stress biomarkers over time, there were interactions for SOD, TBARS, and GPx.
- Specifically, TBARS remained at baseline levels during most of the collection times within weeks 1, 7, and 8. A notable spike was observed on the second day in the second week of training (post-exercise) and remained elevated on the third day (pre- and immediately post-exercise). GPx activity exhibited a similar pattern.
- In contrast, SOD remained at the baseline level until it increased around 30 minutes after exercise on Day 1 of Week 1 and at most collection times of Day 3 in Week 8.
- Moreover, there was a significant drop in concentrations of amino acids that demonstrate antioxidant properties (Met, Tyr, and Trp) from week 2 to 8 of the exercise protocol.
Implications
- The performance of horses is subject to various stressors that can impact their performance. One of the stressors highlighted in this study is exercise.
- This study unveils the time-dependent nature of oxidative stress related to persistent stressors like exercise. The changes over time in the oxidative stress biomarkers and amino acids in blood plasma give an insight into how horses are adapting to their training regimen.
- Better understanding of an athlete horse’s response to continual stressors is important for the design of effective training methods to prevent oxidative damage, optimize performance, and maintain optimal horse health.
Cite This Article
APA
Ott EC, Cavinder CA, Wang S, Smith T, Lemley CO, Dinh TTN.
(2022).
Oxidative stress biomarkers and free amino acid concentrations in the blood plasma of moderately exercised horses indicate adaptive response to prolonged exercise training.
J Anim Sci, 100(4), skac086.
https://doi.org/10.1093/jas/skac086 Publication
Researcher Affiliations
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
MeSH Terms
- Amino Acids / metabolism
- Animals
- Antioxidants / metabolism
- Biomarkers / metabolism
- Glutathione Peroxidase / metabolism
- Horses
- Oxidative Stress / physiology
- Physical Conditioning, Animal / physiology
- Plasma / metabolism
- Superoxide Dismutase / metabolism
- Thiobarbituric Acid Reactive Substances
References
This article includes 40 references
- Andriichuk A, Tkachenko H, Tkachova I. Oxidative stress biomarkers and erythrocytes hemolysis in well-trained equine athletes before and after exercise. J. Equine Vet. Sci. 2016;36:32–43.
- Austin L, Arthur H, de Niese M, Gurusinghe A, Baker MS. Micromethods in single muscle fibers. 2. Determination of glutathione reductase and glutathione peroxidase.. Anal Biochem 1988 Nov 1;174(2):575-9.
- Balogh N, Gaál T, Ribiczeyné PS, Petri A. Biochemical and antioxidant changes in plasma and erythrocytes of pentathlon horses before and after exercise.. Vet Clin Pathol 2001;30(4):214-218.
- Barja G. Mitochondrial oxygen radical generation and leak: sites of production in states 4 and 3, organ specificity, and relation to aging and longevity.. J Bioenerg Biomembr 1999 Aug;31(4):347-66.
- Bowtell JL, Marwood S, Bruce M, Constantin-Teodosiu D, Greenhaff PL. Tricarboxylic acid cycle intermediate pool size: functional importance for oxidative metabolism in exercising human skeletal muscle.. Sports Med 2007;37(12):1071-88.
- Chen Z, Zhong C. Oxidative stress in Alzheimer's disease.. Neurosci Bull 2014 Apr;30(2):271-81.
- Davies KJ, Quintanilha AT, Brooks GA, Packer L. Free radicals and tissue damage produced by exercise.. Biochem Biophys Res Commun 1982 Aug 31;107(4):1198-205.
- Deaton CM, Marlin DJ. Exercise-associated oxidative stress. Clin. Tech. Equine Pract. 2003;2:278–291.
- Draper HH, Squires EJ, Mahmoodi H, Wu J, Agarwal S, Hadley M. A comparative evaluation of thiobarbituric acid methods for the determination of malondialdehyde in biological materials.. Free Radic Biol Med 1993 Oct;15(4):353-63.
- Fisher-Wellman K, Bloomer RJ. Acute exercise and oxidative stress: a 30 year history.. Dyn Med 2009 Jan 13;8:1.
- Fukai T, Ushio-Fukai M. Superoxide dismutases: role in redox signaling, vascular function, and diseases.. Antioxid Redox Signal 2011 Sep 15;15(6):1583-606.
- Hackl S, van den Hoven R, Zickl M, Spona J, Zentek J. The effects of short intensive exercise on plasma free amino acids in standardbred trotters.. J Anim Physiol Anim Nutr (Berl) 2009 Apr;93(2):165-73.
- Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. .
- Hargreaves BJ, Kronfeld DS, Waldron JN, Lopes MA, Gay LS, Saker KE, Cooper WL, Sklan DJ, Harris PA. Antioxidant status of horses during two 80-km endurance races.. J Nutr 2002 Jun;132(6 Suppl 2):1781S-3S.
- Henneke DR, Potter GD, Kreider JL, Yeates BF. Relationship between condition score, physical measurements and body fat percentage in mares.. Equine Vet J 1983 Oct;15(4):371-2.
- Ji LL. Modulation of skeletal muscle antioxidant defense by exercise: Role of redox signaling.. Free Radic Biol Med 2008 Jan 15;44(2):142-52.
- Kaspar H, Dettmer K, Gronwald W, Oefner PJ. Automated GC-MS analysis of free amino acids in biological fluids.. J Chromatogr B Analyt Technol Biomed Life Sci 2008 Jul 15;870(2):222-32.
- Lamprecht ED, Williams CA. Biomarkers of antioxidant status, inflammation, and cartilage metabolism are affected by acute intense exercise but not superoxide dismutase supplementation in horses.. Oxid Med Cell Longev 2012;2012:920932.
- Leeuwenburgh C, Fiebig R, Chandwaney R, Ji LL. Aging and exercise training in skeletal muscle: responses of glutathione and antioxidant enzyme systems.. Am J Physiol 1994 Aug;267(2 Pt 2):R439-45.
- Marlin DJ, Fenn K, Smith N, Deaton CD, Roberts CA, Harris PA, Dunster C, Kelly FJ. Changes in circulatory antioxidant status in horses during prolonged exercise.. J Nutr 2002 Jun;132(6 Suppl 2):1622S-7S.
- de Moffarts B, Kirschvink N, Art T, Pincemail J, Michaux C, Cayeux K, Defraigne JO, Lekeux P. Impact of training and exercise intensity on blood antioxidant markers in healthy standardbred horses. Equine Comp. Exerc. Physiol. 2004;1:211–220.
- NRC. Nutrient requirements of horses. 2007;6th rev. ed..
- Niedzwiedz A, Jaworski Z. Oxidant-antioxidant status in the blood of horses with symptomatic recurrent airway obstruction (RAO).. J Vet Intern Med 2014 Nov-Dec;28(6):1845-52.
- Niedźwiedź A, Kubiak K, Nicpoń J. Plasma total antioxidant status in horses after 8-hours of road transportation.. Acta Vet Scand 2013 Aug 14;55(1):58.
- Ono K, Inui K, Hasegawa T, Matsuki N, Watanabe H, Takagi S, Hasegawa A, Tomoda I. The changes of antioxidative enzyme activities in equine erythrocytes following exercise.. Nihon Juigaku Zasshi 1990 Aug;52(4):759-65.
- Powers SK, Jackson MJ. Exercise-induced oxidative stress: cellular mechanisms and impact on muscle force production.. Physiol Rev 2008 Oct;88(4):1243-76.
- Powers SK, Ji LL, Leeuwenburgh C. Exercise training-induced alterations in skeletal muscle antioxidant capacity: a brief review.. Med Sci Sports Exerc 1999 Jul;31(7):987-97.
- Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay.. Free Radic Biol Med 1999 May;26(9-10):1231-7.
- Reid MB. Invited Review: redox modulation of skeletal muscle contraction: what we know and what we don't.. J Appl Physiol (1985) 2001 Feb;90(2):724-31.
- Reid MB, Khawli FA, Moody MR. Reactive oxygen in skeletal muscle. III. Contractility of unfatigued muscle.. J Appl Physiol (1985) 1993 Sep;75(3):1081-7.
- Sachdev S, Davies KJ. Production, detection, and adaptive responses to free radicals in exercise.. Free Radic Biol Med 2008 Jan 15;44(2):215-23.
- Sen CK. Oxidants and antioxidants in exercise.. J Appl Physiol (1985) 1995 Sep;79(3):675-86.
- Smarsh DN, Williams CA. Oxidative stress and antioxidant status in standardbreds: effect of age and training in resting plasma and muscle. J. Equine Vet. Sci. 2017;48:93–102.
- Somani SM, Frank S, Rybak LP. Responses of antioxidant system to acute and trained exercise in rat heart subcellular fractions.. Pharmacol Biochem Behav 1995 Aug;51(4):627-34.
- Steinbacher P, Eckl P. Impact of oxidative stress on exercising skeletal muscle.. Biomolecules 2015 Apr 10;5(2):356-77.
- Traustadóttir T, Davies SS, Su Y, Choi L, Brown-Borg HM, Roberts LJ 2nd, Harman SM. Oxidative stress in older adults: effects of physical fitness.. Age (Dordr) 2012 Aug;34(4):969-82.
- Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease.. Int J Biochem Cell Biol 2007;39(1):44-84.
- Wagenmakers AJ. Protein and amino acid metabolism in human muscle.. Adv Exp Med Biol 1998;441:307-19.
- Williams CA, Carlucci SA. Oral vitamin E supplementation on oxidative stress, vitamin and antioxidant status in intensely exercised horses.. Equine Vet J Suppl 2006 Aug;(36):617-21.
- Xu N, Chen G, Liu H. Antioxidative Categorization of Twenty Amino Acids Based on Experimental Evaluation.. Molecules 2017 Nov 27;22(12).
Citations
This article has been cited 10 times.- Bollinger L, Bartel A, Weber C, Gehlen H. Pre-Ride Biomarkers and Endurance Horse Welfare: Analyzing the Impact of the Elimination of Superoxide Dismutase, δ-Aminolevulinic-Dehydratase, Thiobarbituric Acid Reactive Substances, Iron, and Serum Amyloid A Levels in Elite 160 km Endurance Rides. Animals (Basel) 2023 May 17;13(10).
- Johnson SE, Barshick MR, Gonzalez ML, Riley JW, Pelletier ME, Castanho BC, Ealy EN. A Carnitine-Containing Product Improves Aspects of Post-Exercise Recovery in Adult Horses. Animals (Basel) 2023 Feb 14;13(4).
- Reemtsma FP, Giers J, Horstmann S, Stoeckle SD, Gehlen H. Evaluation of Concentration Changes in Plasma Amino Acids and Their Metabolites in Eventing Horses During Cross-Country Competitions as Potential Performance Predictors. Animals (Basel) 2025 Dec 17;15(24).
- Li F, Li S, Xie H. The Effects of Physical Activity on Skin Health: A Narrative Review. Clin Cosmet Investig Dermatol 2025;18:2189-2200.
- Reemtsma FP, Giers J, Horstmann S, Stoeckle SD, Gehlen H. Concentration Changes in Plasma Amino Acids and Their Metabolites in Eventing Horses During Cross-Country Competitions. Animals (Basel) 2025 Jun 22;15(13).
- James C, Lloyd EM, Arthur PG. The Level of Thiol-Oxidised Plasma Albumin Is Elevated Following a Race in Australian Thoroughbred Horses. Vet Med Sci 2025 Jul;11(4):e70487.
- Barshick MR, Ely KM, Mogge KC, Chance LM, Johnson SE. Methylsulfonylmethane (MSM) Supplementation in Adult Horses Supports Improved Skeletal Muscle Inflammatory Gene Expression Following Exercise. Animals (Basel) 2025 Jan 14;15(2).
- Aitken RJ, Wilkins A, Harrison N, Bahrami M, Gibb Z, McIntosh K, Vuong Q, Lambourne S. A Comparative Analysis of the Antioxidant Profiles Generated by the RoXsta(TM) System for Diverse Biological Fluids Highlights the Powerful Protective Role of Human Seminal Plasma. Antioxidants (Basel) 2025 Jan 14;14(1).
- Giercuszkiewicz-Hecold B, Kulka M, Czopowicz M, Wilczak J, Szarska E, Strzelec K, Grzeczka A, Graczyk S, Hryniszyn A, Mularczyk M, Marycz K, Cywińska A. The effect of long term astaxanthin supplementation on the antioxidant status of racing Arabian horses - preliminary study. Sci Rep 2024 Nov 14;14(1):27991.
- Hisaeda K, Ono T, Kadekaru S, Hata A, Miyama TS, Kutara K, Sugimoto K, Hiasa Y, Ohzawa E, Kunieda T, Iwata E, Kitagawa H. Serum amino acid profiles in clinically normal Noma horses. J Equine Sci 2024;35(2):29-34.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists