European Journal of Applied Physiology

, Volume 95, Issue 5–6, pp 550–556

Oxygen radical absorbance capacity (ORAC) and exercise-induced oxidative stress in trotters

  • Susanna Kinnunen
  • Seppo Hyyppä
  • Arja Lehmuskero
  • Niku Oksala
  • Pekka Mäenpää
  • Osmo Hänninen
  • Mustafa Atalay
Original Article


Strenuous exercise is a potent inducer of oxidative stress, which has been suggested to be associated with disturbances in muscle homeostasis, fatigue and injury. There is no comprehensive or uniform view of the antioxidant status in horses. We have previously shown that moderate exercise induces protein oxidation in trotters. The aim of this study was to measure the antioxidative capacity of the horse in relation to different antioxidant components and oxidative stress markers after a single bout of moderate exercise to elucidate the mechanisms of antioxidant protection in horses. Eight clinically normal and regularly trained standard-bred trotters were treadmill-exercised for 53 min at moderate intensity. Blood samples were collected prior to and immediately after exercise and at 4 and 24 h of recovery. Muscle biopsies from the middle gluteal muscle were taken before exercise and after 4 h of recovery. Acute induction of oxygen radical absorbance capacity (ORAC) did not prevent exercise-induced oxidative stress, which was demonstrated by increased lipid hydroperoxides (LPO). Pre-exercise ORAC levels were, however, a determinant of total glutathione content of the blood after 4 and 24 h of recovery. Furthermore, baseline ORAC level correlated negatively with 4-h recovery LPO levels. Our results imply that horses are susceptible to oxidative stress, but a stronger antioxidant capacity may improve coping with exercise-induced oxidative stress.


Exercise Horse Oxidative stress Recovery Total antioxidant capacity 


  1. Alessio HM, Hagerman AE, Fulkerson BK, Ambrose J, Rice RE, Wiley RL (2000) Generation of reactive oxygen species after exhaustive aerobic and isometric exercise. Med Sci Sports Exerc 32:1576–1581PubMedCrossRefGoogle Scholar
  2. Arab K, Steghens JP (2004) Plasma lipid hydroperoxides measurement by an automated xylenol orange method. Anal Biochem 325:158–163PubMedCrossRefGoogle Scholar
  3. Atalay M, Laaksonen DE, Khanna S, Kaliste-Korhonen E, Hänninen O, Sen CK (2000) Vitamin E regulates changes in tissue antioxidants induced by fish oil and acute exercise. Med Sci Sports Exerc 32:601–607PubMedCrossRefGoogle Scholar
  4. Atalay M, Laaksonen DE (2002) Diabetes, oxidative stress and physical exercise. Journal of Sports Science and Medicine 1:1–14Google Scholar
  5. Atalay M, Oksala NK, Laaksonen DE, Khanna S, Nakao C, Lappalainen J, Roy S, Hänninen O, Sen CK (2004) Exercise training modulates heat shock protein response in diabetic rats. J Appl Physiol 97:605–611PubMedCrossRefGoogle Scholar
  6. Avellini L, Silvestrelli M, Gaiti A (1995) Training-induced modifications in some biochemical defences against free radicals in equine erythrocytes. Vet Res Commun 19:179–184PubMedCrossRefGoogle Scholar
  7. Balakrishnan VS, Blumberg J, Pereira BJ, Jaber BL (2003) Antioxidant and oxidative stress indices in dialysis-dependent acute renal failure. Blood Purif 21:213–219PubMedCrossRefGoogle Scholar
  8. Balogh N, Gaal T, Ribiczeyne PS, Petri A (2001) Biochemical and antioxidant changes in plasma and erythrocytes of pentathlon horses before and after exercise. Vet Clin Pathol 30:214–218PubMedCrossRefGoogle Scholar
  9. Cao G, Alessio HM, Cutler RG (1993) Oxygen-radical absorbance capacity assay for antioxidants. Free Radic Biol Med 14:303–311PubMedCrossRefGoogle Scholar
  10. Cao G, Shukitt-Hale B, Bickford PC, Joseph JA, McEwen J, Prior RL (1999) Hyperoxia-induced changes in antioxidant capacity and the effect of dietary antioxidants. J Appl Physiol 86:1817–1822PubMedCrossRefGoogle Scholar
  11. Childs A, Jacobs C, Kaminski T, Halliwell B, Leeuwenburgh C (2001) Supplementation with vitamin C and N-acetyl-cysteine increases oxidative stress in humans after an acute muscle injury induced by eccentric exercise. Free Radic Biol Med 31:745–753PubMedCrossRefGoogle Scholar
  12. De Leenheer AP, De Bevere VO, De Ruyter MG, Claeys AE (1979) Simultaneous determination of retinol and alpha-tocopherol in human serum by high-performance liquid chromatography. J Chromatogr 162:408–413PubMedCrossRefGoogle Scholar
  13. Gul M, Atalay M, Hänninen O (2003) Endurance training and glutathione-dependent antioxidant defense mechanism in heart of the diabetic rats. Journal of Sports Science and Medicine 2:52–61Google Scholar
  14. Jackson MJ, Papa S, Bolanos J, Bruckdorfer R, Carlsen H, Elliott RM, Flier J, Griffiths HR, Heales S, Holst B, Lorusso M, Lund E, Oivind Moskaug J, Moser U, Di Paola M, Polidori MC, Signorile A, Stahl W, Vina-Ribes J, Astley SB (2002) Antioxidants, reactive oxygen and nitrogen species, gene induction and mitochondrial function. Mol Aspects Med 23:209–285PubMedCrossRefGoogle Scholar
  15. Jiao H, Wang SY (2000) Correlation of antioxidant capacities to oxygen radical scavenging enzyme activities in blackberry. J Agric Food Chem 48:5672–5676PubMedCrossRefGoogle Scholar
  16. Kaikkonen J, Porkkala-Sarataho E, Tuomainen TP, Nyyssönen K, Kosonen L, Ristonmaa U, Lakka HM, Salonen R, Korpela H, Salonen JT (2002) Exhaustive exercise increases plasma/serum total oxidation resistance in moderately trained men and women, whereas their VLDL + LDL lipoprotein fraction is more susceptible to oxidation. Scand J Clin Lab Invest 62:599–607PubMedCrossRefGoogle Scholar
  17. Kayatekin BM, Gonenc S, Acikgoz O, Uysal N, Dayi A (2002) Effects of sprint exercise on oxidative stress in skeletal muscle and liver. Eur J Appl Physiol 87:141–144PubMedCrossRefGoogle Scholar
  18. Kinnunen S, Hyyppä S, Lappalainen J, Oksala N, Venojärvi M, Nakao C, Hänninen O, Sen CK, Atalay M (2005) Exercise-induced oxidative stress and muscle stress protein responses in trotters. Eur J Appl Physiol 93:496–501PubMedCrossRefGoogle Scholar
  19. Laaksonen DE, Atalay M, Niskanen L, Uusitupa M, Hänninen O, Sen CK (1999) Blood glutathione homeostasis as a determinant of resting and exercise-induced oxidative stress in young men. Redox Rep 4:53–59PubMedCrossRefGoogle Scholar
  20. Lee J, Goldfarb AH, Rescino MH, Hegde S, Patrick S, Apperson K (2002) Eccentric exercise effect on blood oxidative-stress markers and delayed onset of muscle soreness. Med Sci Sports Exerc 34:443–448PubMedCrossRefGoogle Scholar
  21. Lindholm A, Piehl K (1974) Fibre composition, enzyme activity and concentrations of metabolites and electrolytes in muscles of standardbred horses. Acta Vet Scand 15:287–309PubMedGoogle Scholar
  22. Mills PC, Smith NC, Casas I, Harris P, Harris RC, Marlin DJ (1996) Effects of exercise intensity and environmental stress on indices of oxidative stress and iron homeostasis during exercise in the horse. Eur J Appl Physiol Occup Physiol 74:60–66PubMedCrossRefGoogle Scholar
  23. Naguib Y, Hari SP, Passwater R, Jr., Huang D (2003) Antioxidant activities of natural vitamin E formulations. J Nutr Sci Vitaminol (Tokyo) 49:217–220Google Scholar
  24. Oztasan N, Taysi S, Gumustekin K, Altinkaynak K, Aktas O, Timur H, Siktar E, Keles S, Akar S, Akcay F, Dane S, Gul M (2004) Endurance training attenuates exercise-induced oxidative stress in erythrocytes in rat. Eur J Appl Physiol 91:622–627PubMedCrossRefGoogle Scholar
  25. Pieri C, Marra M, Moroni F, Recchioni R, Marcheselli F (1994) Melatonin: a peroxyl radical scavenger more effective than vitamin E. Life Sci 55:PL271–276PubMedCrossRefGoogle Scholar
  26. Prior RL, Cao G (1999) In vivo total antioxidant capacity: comparison of different analytical methods. Free Radic Biol Med 27:1173–1181PubMedCrossRefGoogle Scholar
  27. Prior RL, Hoang H, Gu L, Wu X, Bacchiocca M, Howard L, Hampsch-Woodill M, Huang D, Ou B, Jacob R (2003) Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORAC(FL))) of plasma and other biological and food samples. J Agric Food Chem 51:3273–3279PubMedCrossRefGoogle Scholar
  28. Räsänen LA, Myllymäki T, Hyyppä S, Maisi P, Pösö AR (1993) Accumulation of allantoin and uric acid in plasma of exercising trotters. Am J Vet Res 54:1923–1928PubMedGoogle Scholar
  29. Räsänen LA, Wiitanen PA, Lilius EM, Hyyppä S, Pösö AR (1996) Accumulation of uric acid in plasma after repeated bouts of exercise in the horse. Comp Biochem Physiol B Biochem Mol Biol 114:139–144PubMedCrossRefGoogle Scholar
  30. Sacheck JM, Milbury PE, Cannon JG, Roubenoff R, Blumberg JB (2003) Effect of vitamin E and eccentric exercise on selected biomarkers of oxidative stress in young and elderly men. Free Radic Biol Med 34:1575–1588PubMedCrossRefGoogle Scholar
  31. Sen CK, Marin E, Kretzschmar M, Hänninen O (1992) Skeletal muscle and liver glutathione homeostasis in response to training, exercise, and immobilization. J Appl Physiol 73:1265–1272PubMedGoogle Scholar
  32. Sen CK, Atalay M, Hänninen O (1994) Exercise-induced oxidative stress: glutathione supplementation and deficiency. J Appl Physiol 77:2177–2187PubMedGoogle Scholar
  33. Sen CK, Atalay M, Å gren J, Laaksonen DE, Roy S, Hänninen O (1997) Fish oil and vitamin E supplementation in oxidative stress at rest and after physical exercise. J Appl Physiol 83:189–195PubMedGoogle Scholar
  34. Sen CK, Packer L (2000) Thiol homeostasis and supplements in physical exercise. Am J Clin Nutr 72:653S–669SPubMedGoogle Scholar
  35. Smith NC, Dunnett M, Mills PC (1995) Simultaneous quantitation of oxidised and reduced glutathione in equine biological fluids by reversed-phase high-performance liquid chromatography using electrochemical detection. J Chromatogr B Biomed Appl 673:35–41PubMedCrossRefGoogle Scholar
  36. Warburton DE, Gledhill N, Jamnik VK, Krip B, Card N (1999) Induced hypervolemia, cardiac function, VO2max, and performance of elite cyclists. Med Sci Sports Exerc 31:800–808PubMedCrossRefGoogle Scholar
  37. White A, Estrada M, Walker K, Wisnia P, Filgueira G, Valdes F, Araneda O, Behn C, Martinez R (2001) Role of exercise and ascorbate on plasma antioxidant capacity in thoroughbred race horses. Comp Biochem Physiol A Mol Integr Physiol 128:99–104PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Susanna Kinnunen
    • 1
    • 2
  • Seppo Hyyppä
    • 3
  • Arja Lehmuskero
    • 2
  • Niku Oksala
    • 1
    • 4
  • Pekka Mäenpää
    • 2
  • Osmo Hänninen
    • 1
  • Mustafa Atalay
    • 1
  1. 1.Department of PhysiologyUniversity of KuopioKuopioFinland
  2. 2.Equine Information CentreKuopioFinland
  3. 3.Agricultural Research CentreEquine ResearchYpäjäFinland
  4. 4.Division of Vascular Surgery, Department of SurgeryTampere University HospitalTampereFinland

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