International Journal of Biometeorology

, Volume 63, Issue 12, pp 1569–1584 | Cite as

Cellular antioxidant enzyme activity and biomarkers for oxidative stress are affected by heat stress

  • Walid S. Habashy
  • Marie C. Milfort
  • Romdhane Rekaya
  • Samuel E. AggreyEmail author
Original Paper


Heat stress (HS) causes oxidative stress and cellular changes in an attempt to detoxify the harmful effects of reactive oxygen species (ROS). However, how ROS affect different organs in chickens under acute and chronic HS is relatively unknown. We investigated the cellular enzyme activity and biomarker changes in the liver and Pectoralis (P) major muscle in broiler chickens subjected to both acute and chronic HS. Forty-eight broiler chickens at 14 days old were randomly assigned to either 25 °C (control) or 35 °C (heat-stressed) for 12 days. Five birds per treatment at 1 and 12 days post-HS were euthanized, and the liver and P. major muscle were sampled. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione (GSH), glutathione reductase (GR), glutathione S-transferase (GST) activity as well as 8-hydroxy-2′-deoxyguanosine (8-OHdG), advanced glycation end product (AGE), malondialdehyde (MDA), and protein carbonyl (PCO) were analyzed as biomarkers for DNA, carbohydrate, lipid, and protein oxidation, respectively. The SOD, CAT, and GSH-GPx activity levels in the liver and the P. major muscle changed under HS; however, some of the changes were tissue-specific or dependent on the duration of the HS. There were increased liver 8-OHdG during chronic HS and also increased liver AGE levels during both acute and chronic HS indicating significant carbohydrate and DNA oxidations. In the P. major muscle, we observed significant increases in lipid peroxidation and protein oxidation which may reflect that this tissue is less resilient to oxidative damage under heat stress. We show that heat stress caused tissue-specific changes to levels of oxidation biomarkers in chicken.


Heat stress Oxidative stress DNA oxidation Lipid peroxidation Protein oxidation 



Walid Habashy was supported by the Missions Sector of the Egyptian Ministry of Higher Education.


  1. Ahmed N (2005) Advanced glycation end products: role in pathology of diabetic complications. Diabetes Res Clin Pract 67:3–21CrossRefGoogle Scholar
  2. Altan O, Pabuccuoglu A, Altan A, Konyalioglu S, Bayraktar H (2003) Effect of heat stress on oxidative stress, lipid peroxidation and some stress parameters in broilers. Br Poult Sci 44:545–550CrossRefGoogle Scholar
  3. Azad MAK, Kikusato M, Maekawa T, Shirakawa H, Toyomizu M (2010) Metabolic characteristics and oxidative damage to skeletal muscle in broiler chickens exposed to chronic heat stress. Comp. Comp Biochem Physiol A Mol Integr Physiol 155:401–406CrossRefGoogle Scholar
  4. Barciszewski J, Barciszewski MZ, Siboska G, Rattan SI, Clark BF (1999) Some unusual nucleic acid bases are products of hydroxyl radical oxidation of DNA and RNA. Mol Biol Rep 26:231–238CrossRefGoogle Scholar
  5. Baud O, Greene AE, Li J, Wang H, Volpe JJ, Rosenberg PA (2004) Glutathione peroxidase–catalase cooperativity is required for resistance to hydrogen peroxide by mature rat oligodendrocytes. J Neurol Sci 24:1531–1540Google Scholar
  6. Bucala R, Cerami A (1992) Advanced glycation end products: role in pathology of diabetic complications. Adv Pharmacol 23:1–34Google Scholar
  7. Cadenas E, Davies KJA (2000) Mitochondrial free radical generation, oxidative stress and aging. Free Radic Biol Med 29:222–230CrossRefGoogle Scholar
  8. Chen J, Bhandar B, Kavdia M (2015) Interaction of ROS and RNS with GSH and GSH/GPX systems. FASEB J 29:636–637CrossRefGoogle Scholar
  9. Cheng JZ, Sharma R, Yang Y, Singhal SS, Sharma A, Saini MK, Singh SV, Zimniak P, Awasthi S, Awasthi YC (2001) Accelerated metabolism and exclusion of 4-hydroxynonenal through induction of RLIP76 and hGST5.8 is an early adaptive response of cells to heat and oxidative stress. J Biol Chem 276:41213–41223CrossRefGoogle Scholar
  10. Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R (2003) Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta 329:23–38CrossRefGoogle Scholar
  11. Dalle-Donne I, Aldini G, Carini M, Colombo R, Rossi R, Milzani A (2006) Protein carbonylation, cellular dysfunction and disease progression. J Cell Mol Med 10:389–406CrossRefGoogle Scholar
  12. Dean RT, Roberts CR, Jessup W (1985) Fragmentation of extracellular and intracellular polypeptides by free radicals. Prog Clin Biol Res 180:341–350Google Scholar
  13. Dröge W (2002) Free radicals in physiological control of cell function. Physiol Rev 82:47–95CrossRefGoogle Scholar
  14. El-Bahr SM (2013) Biochemistry of free radicals and oxidative stress. Sci Int 1:111–117CrossRefGoogle Scholar
  15. Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408:239–247CrossRefGoogle Scholar
  16. Flohé L (2013) The fairytale of the GSSG/GSH redox potential. Biochim Biophys Acta 1830:3139–3142CrossRefGoogle Scholar
  17. Fridovich I (1997) Superoxide anion radical, superoxide dismutases, and related matters. J Biol Chem 272:18515–18517CrossRefGoogle Scholar
  18. Geraert PA, Padilha CF, Guillaumin S (1996) Metabolic and endocrine changes induced by chronic heat exposure in broiler chickens: growth performance, body composition and energy retention. Br J Nutr 75:195–204Google Scholar
  19. Girotti AW (1985) Mechanisms of lipid peroxidation. J Free Radic Biol Med 1:87–95CrossRefGoogle Scholar
  20. Habashy WS, Milfort MC, Adomako K, Attia YA, Rekaya R, Aggrey SE (2017a) Effect of heat stress on amino acid digestibility and transporters in meat-type chickens. Poult Sci 96:2312–2319CrossRefGoogle Scholar
  21. Habashy WS, Milfort MC, Fuller AL, Attia YA, Rekaya R, Aggrey SE (2017b) Effect of heat stress on protein utilization and nutrient transporters in meat-type chickens. Int J Biometeorol 61:2111–2118CrossRefGoogle Scholar
  22. Habashy WS, Milfort MC, Rekaya R, Aggrey SE (2018) Expression of genes that encode cellular oxidant/antioxidant systems are affected by heat stress. Mol Biol Rep 45:389–394CrossRefGoogle Scholar
  23. Halliwell B (2001) Free radicals and other reactive species in disease. Encyclopedia of life sciences. Nature Publishing Group / Accessed 15 Dec 2018
  24. Hayes JD, McLellan LI (1999) Glutathione and glutathione-dependent enzymes represent a co-ordinately regulated defence against oxidative stress. Free Radic Res 31:273–300CrossRefGoogle Scholar
  25. Hayes JD, Flanagan JU, Jowsey IR (2005) Glutathione transferases. Annu Rev Pharmacol Toxicol 45:51–88CrossRefGoogle Scholar
  26. Helbock HJ, Beckman KB, Ames BN (1999) 8-hydroxydeoxyguanosine and 8-hydroxyguanine as biomarkers of oxidative DNA damage. Methods Enzymol 300:156–166CrossRefGoogle Scholar
  27. Huang C, Jiao H, Song Z, Zhao J, Wang X, Lin H (2015) Heat stress impairs mitochondria functions and induces oxidative injury in broiler chickens. J Anim Sci 93:2144–2153CrossRefGoogle Scholar
  28. Ismail IB, Al-Busadah KA, El-Bahr SM (2013) Oxidative stress biomarkers and biochemical profile in broilers chicken fed zinc bacitracin and ascorbic acid under hot climate. Am J Biochem Mol Biol 3:202–214CrossRefGoogle Scholar
  29. Jenkins AJ, Lyons TF (1997) Glycation, oxidation, and lipoxidation in the development of the complications of diabetes: a carbonyl stress hypothesis. Diabetes Rev (Alex) 5:365–391Google Scholar
  30. Kelly FJ, Mudway IS (2003) Protein oxidation at the air-lung interface. Amino Acids 25:375–396CrossRefGoogle Scholar
  31. Kikusato M, Toyomizy M (2013) Crucial role of membrane potential in heat stress-induced overproduction of reactive oxygen species in avian skeletal muscle mitochondria. PLoS One 8:e64412CrossRefGoogle Scholar
  32. Kojima T, Norose T, Tsuchiya K, Sakamoto K (2010) Mouse 3T3-L1 cells acquire resistance against oxidative stress as the adipocytes differentiate via the transcription factor Foxo. Apoptosis 15:83–93CrossRefGoogle Scholar
  33. Kuehe A, Emmert H, Soehle J, Winnefeld M, Fischer F, Wenck H, Gallinat S, Terstegen L, Lucius R, Hildebrand J, Zamboni N (2015) Acute activation of oxidative pentose phosphate pathway as first-line response to oxidative stress in human skin cells. Mol Cell 59:359–371CrossRefGoogle Scholar
  34. Levine RL, Williams JA, Stadtman ER, Shacter E (1994) Carbonyl assays for determination of oxidatively modified proteins. Methods Enzymol 233:346–357CrossRefGoogle Scholar
  35. Levine RL, Moskovitz J, Stadtman ER (2000) Oxidation of methionine in proteins: roles in antioxidant defense and cellular regulation. IUBMB Life 50:301–307CrossRefGoogle Scholar
  36. Lin H, Du R, Zhang ZY (2000) Peroxide status in tissues of heat stressed broilers. Asian-Australas J Anim Sci 13:1373–1376CrossRefGoogle Scholar
  37. Lu T, Piao XL, Zhang Q, Wang D, Piao XS, Kim SW (2010) Protective effects of Forsythia suspensa extract against oxidative stress induced by diquat in rats. Food Chem Toxicol 48:764–770CrossRefGoogle Scholar
  38. Lyras L, Cairns NJ, Jenner A, Jenner P, Halliwell B (1997) An assessment of oxidative damage to proteins, lipids, and DNA in brain from patients with Alzheimer’s disease. J Neurochem 68:2061–2069CrossRefGoogle Scholar
  39. Mashaly MM, Hendricks GL 3rd, Kalama MA, Gehad AE, Abbas AO, Patterson PH (2004) Effect of heat stress on production parameters and immune responses of commercial laying hens. Poult Sci 83:889–894CrossRefGoogle Scholar
  40. Mujahid A, Akiba Y, Toyomizu M (2007) Acute heat stress induces oxidative stress and decrease adaptation in young White Leghorn cockerels by down regulation of avian uncoupling protein. Poult Sci 86:364–371CrossRefGoogle Scholar
  41. Mujahid A, Akiba Y, Toyomizu M (2009) Progressive changes in the physiological responses of heat-stressed broiler chickens. J Poult Sci 46:163–167CrossRefGoogle Scholar
  42. Murphy ME, Kehrer JP (1989) Oxidation state of tissue thiol groups and content of protein carbonyl groups in chickens with inherited muscular dystrophy. Biochem J 260:359–364CrossRefGoogle Scholar
  43. Ramnath V, Rekha PS, Sujatha KS (2008) Amelioration of heat stress induced disturbances of antioxidant defense system in chicken by Brahma Rasayana. Evid Based Complement Alternat Med 5:77–84CrossRefGoogle Scholar
  44. Rozenboim I, Tako E, Gal-Garber O, Proudman JA, Uni Z (2007) The effect of heat stress on ovarian function of laying hens. Poult Sci 86:1760–1765CrossRefGoogle Scholar
  45. Sahin K, Orhan C, Tuzcu M, Ali S, Sahin N, Hayirli A (2010) Epigallocatechin-3-gallate prevents lipid peroxidation and enhances antioxidant defense system via modulating hepatic nuclear transcription factors in heat-stressed quails. Poult Sci 89:2251–2258CrossRefGoogle Scholar
  46. SAS Institute (2011) SAS/ SAT user guide: statistics ver, 9.3 edn. AS Inst. Inc., CaryGoogle Scholar
  47. Seven PT, Yilmaz S, Seven I, Cerci IH, Azman MA, Yilmaz M (2009) Effects of propolis on selected blood indicators and antioxidant enzyme activities in broilers under heat stress. Acta Vet Brno 78:75–83CrossRefGoogle Scholar
  48. Siu GM, Draper HH (1982) Metabolism of malonaldehyde in vivo and in vitro. Lipids 17:349–355CrossRefGoogle Scholar
  49. Srinivasan U, Mieyal PA, Mieyal JJ (1997) pH profiles indicative of rate limiting nucleophilic displacement in thioltransferase catalysis. Biochem 36:3199–3206CrossRefGoogle Scholar
  50. Sultana R, Cenini G, Butterfield DA (2013) Biomarkers of oxidative stress in neurodegenerative diseases. In: Villamena FA (ed) Molecular basis of oxidative stress: chemistry, mechanisms, and disease pathogenesis. John Wiley and Sons, Inc., New York, pp 359–376CrossRefGoogle Scholar
  51. Sun X, Zhang H, Sheikhahmadi A, Wang Y, Jiao H, Lin H, Song Z (2015) Effects of heat stress on the gene expression of nutrient transporters in the jejunum of broiler chickens (Gallus gallus domesticus). Int J Biometeorol 59:127–113CrossRefGoogle Scholar
  52. Tan GY, Yang L, Fu YQ, Feng JH, Zhang MH (2010) Effect of different acute ambient temperatures on function of hepetic mitochondria respiration, antioxidative enzymes, and oxidative injury in broiler chickens. Poult Sci 89:115–122CrossRefGoogle Scholar
  53. Temim S, Chagneau AM, Guillaumin S, Michel J, Peresson R, Geraert PA, Tesseraud S (1999) Effects of chronic heat exposure and protein intake on growth performance, nitrogen retention and muscle development in broiler chickens. Reprod Nutr Dev 39:145–156CrossRefGoogle Scholar
  54. Thannickal VJ, Fanburg BL (2000) Reactive oxygen species in cell signaling. Am J Phys Lung Cell Mol Phys 279:L1005–L1028Google Scholar
  55. Wautier M, Chappey O, Corda S, Stern DM, AM, Wautier JL (2001) Activation of NADPH oxidase by AGE links oxidant stress to altered gene expression via RAGE. Am J Physiol Endocrinol Metab 280:E685–E694CrossRefGoogle Scholar
  56. Yang L, Tan GY, Fu YQ, Feng JH, Zhang MH (2010) Effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration ROS production and lipid peroxidation in broiler chickens. Comp Biochem Physiol C Toxicol Pharmacol 151:204–208CrossRefGoogle Scholar
  57. Zhang X, Wu RS, Fu W, Xu L, Lam PK (2004) Production of reactive oxygen species and 8-hydroxy-2'deoxyguanosine in KB cells co-exposed to benzo[a]pyrene and UV-A radiation. Chemosphere 55:1303–1308CrossRefGoogle Scholar
  58. Zhang W, Ahn DU, Lee EJ, Xiao S (2011) Consumption of oxidized oil increases oxidative stress in broilers and affects the quality of breast meat. J Agric Food Chem 59:969–974CrossRefGoogle Scholar
  59. Zhang W, Xiao S, Ahn DU (2013) Protein oxidation: basic principles and implications for meat quality. Crit Rev Food Sci Nutr 53:1191–1201CrossRefGoogle Scholar
  60. Zhu H, Wang J, Santo A, Li Y (2013) Downregulation of antioxidants and phase 2 proteins. In: Villamena FA (ed) Molecular basis of oxidative stress: chemistry, mechanisms, and disease pathogenesis. John Wiley and Sons, Inc., New York, pp 113–121CrossRefGoogle Scholar
  61. Zitka O, Skalickova S, Gumulec J, Masarik M, Adam V, Hubalek J, Trnkova L, Kruseova J, Eckschlager T, Kizek R (2012) Redox status expressed as GSH:GSSG ratio as a marker for oxidative stress in paediatric tumour patients. Oncol Lett 4:1247–1253CrossRefGoogle Scholar

Copyright information

© ISB 2019

Authors and Affiliations

  1. 1.NutriGenomics Laboratory, Department of Poultry ScienceUniversity of GeorgiaAthensUSA
  2. 2.Department of Animal and Poultry ProductionDamanhour UniversityDamanhourEgypt
  3. 3.Department of Animal and Dairy ScienceUniversity of GeorgiaAthensUSA

Personalised recommendations