Fish Physiology and Biochemistry

, Volume 44, Issue 3, pp 939–948 | Cite as

Effects of dietary inclusions of red beet and betaine on the acute stress response and muscle lipid peroxidation in rainbow trout

  • Julia Pinedo-Gil
  • Ana Belén Martín-Diana
  • Daniela Bertotto
  • Miguel Ángel Sanz-Calvo
  • Miguel Jover-Cerdá
  • Ana Tomás-Vidal


This study evaluates the effects of red beet (RB) and betaine on rainbow trout submitted to an acute stress challenge. A control diet was compared with four experimental diets in which red beet (14 and 28%) and betaine (0.9 and 1.63%) were incorporated in different concentrations according to a factorial design. Cortisol in plasma and fin, glucose and lactate plasma levels, and malondialdehide (MDA) in muscle were all measured before the stress challenge and 30 min and 6 and 12 h after the stress challenge as parameters to determine the diet effects. RB and betaine had no effect on cortisol, glucose, and MDA basal levels. However, lactate basal levels were significantly lower on fish fed with RB and betaine. Thirty minutes after the stress challenge, there was a significant increase in plasma and fin cortisol, glucose and lactate concentrations, although fish fed with diets containing RB and betaine showed significantly higher plasma cortisol values. MDA values of fish fed with 14% RB and 0.9% betaine were significantly higher than MDA values from fish fed with 28% RB and 1.63% betaine. After 6 and 12 h, plasma and fin cortisol and lactate levels recovered in a similar trend. Glucose plasma levels recovered in almost all groups 12 h after the stress. Also, MDA values recovered basal levels after 6 and 12 h. RB and betaine did not enhance the tolerance to the stress challenge compared to the control group, although the presence of these ingredients had no negative effect on any of the stress indicators.


Red beet Betaine Rainbow trout Acute stress challenge 



This work has been co-funded with FEDER and INIA funds. Julia Pinedo has been granted with the FPI-INIA grant number 21 (call 2012, BOE-2012-13337).

Compliance with ethical standards

Ethical statement

The rainbow trout Oncorhynchus mykiss (Walbaum) study complied with the European Union Council 282 Directive 2010/63/UE, which lays down minimum standards for the protection of animals, and Spanish national legislation (Spanish Royal Decree 53/2013) protecting animals used in experimentation and for other scientific purposes and approved by Animal Ethics Committee of Agro-Technological Institute of Castilla y León (Spain).


  1. Aluru N, Vijayan MM (2006) Aryl hydrocarbon receptor activation impairs cortisol response to stress in rainbow trout by disrupting the rate limiting steps in steroidogenesis. Endocrinology 147:1895–1903CrossRefPubMedGoogle Scholar
  2. Ashley PJ (2007) Fish welfare: current issues in aquaculture. Appl Anim Behav Sci 104:199–235CrossRefGoogle Scholar
  3. Barton BA, Iwama GK (1991) Physiological changes in fish from stress in aquaculture with emphasis on the response and effects of corticosteroids. Annu Rev Fish Dis 1:3–26CrossRefGoogle Scholar
  4. Bertotto D, Poltronieri C, Negrato E, Majolini D, Radaelli G, Simontacchi C (2010) Alternative matrices for cortisol measurement in fish. Aquac Res 41:1261–1267Google Scholar
  5. Bertotto D, Poltronieri C, Negrato E, Richard J, Pascoli F, Simontacchi C, Radaelli G (2011) Whole body cortisol and expression of HSP70, IGF-I and MSTN in early development of sea bass subjected to heat shok. Gen Comp Endocrinol 174:44–50CrossRefPubMedGoogle Scholar
  6. Chagas EC, Val AL (2006) Ascorbic acid reduces the effects of hypoxia on the Amazon fish tambaqui. J Fish Biol 69:608–612CrossRefGoogle Scholar
  7. Cui XJ, Zhou QC, Liang HO, Yang J, Zhao LM (2010) Effects of dietary carbohydrate sources on the growth performance and hepatic carbohydrate metabolic enzyme activities of juvenile cobia (Rachycentron canadum Linnaeus.) Aquac Res 42:99–107CrossRefGoogle Scholar
  8. Dabrowski K, Lee KJ, Guz L, Verlhac V, Gabaudan J (2004) Effects of dietary ascorbic acid on oxygen stress (hypoxia or hyperoxia), growth and tissue vitamin concentration in juvenile rainbow trout (Oncorhynchus mykiss). Aquaculture 233:383–392CrossRefGoogle Scholar
  9. Enes P, Panserat S, Kaushik S, Oliva-Teles A (2006) Rapid metabolic adaptation of European sea beass (Dicentrarchus labrax) juveniles fed different carbohydrate sources after heat shock stress. Comp Biochem Physiol A 145:73–81CrossRefGoogle Scholar
  10. Fast MD, Hosoya S, Johnson SC, Alfonso LOB (2008) Cortisol response and immune-related effects of Atlantic salmon (Salmo salar Linnaeus) subjected to short- and long-term stress. Fish Shellfish Immunol 24:194–204CrossRefPubMedGoogle Scholar
  11. Francis G, Makkar HPS, Becker K (2001) Antinutritional factors present in plant-derived alternate fish feed ingredients and their effects in fish. Aquaculture 199:197–227CrossRefGoogle Scholar
  12. Ganessan B, Anandan R, Lakshmanan PT (2011) Studies on the protective effects of betaine against oxidative damage during experimentally induced restraint stress in Wistar albino rats. Cell Stress Chaperones 16:641–652CrossRefGoogle Scholar
  13. Gesto M, López-Patiño MA, Hernández J, Soengas JL, Míguez JM (2013) The response of brain serotonergic and dopaminergic systems to an acute stressor in rainbow trout: a time course study. J Exp Biol 216:4435–4442CrossRefPubMedGoogle Scholar
  14. Gesto M, López-Patiño MA, Hernández J, Soengas JL, Míguez JM (2015) Gradation of the stress response in rainbow trout exposed to stressors of different severity: the role of brain serotonergic and dopaminergic systems. J Neuroendocrinol 27:131–141CrossRefPubMedGoogle Scholar
  15. Hemre GI, Mommsen TP, Krogdahl Å (2002) Carbohydrates in fish nutrition: effects on growth, glucose metabolism and heptic enzymes. Aquac Nutr 8:175–194CrossRefGoogle Scholar
  16. Ings JS, Vijayan MM, Servos MR (2012) Tissue-specific metabolic changes in response to an acute handling disturbance in juvenile rainbow trout exposed to municipal wastewater effluent. Aquat Toxicol 108:53–59CrossRefPubMedGoogle Scholar
  17. Janssens PA, Waterman J (1988) Hormonal regulation of gluconeogenesis and glycogenolysis in carp (Cyprinus carpio) liver pieces cultured in vitro. Comp Biochem Physiol 91A:451–457CrossRefGoogle Scholar
  18. Jeney G, Galeotti M, Volpatti D, Anderson DP (1997) Prevention of stress in rainbow trout (Oncorhynchus mykiss) fed diets containing different doses of glucan. Aquaculture 154:1–15CrossRefGoogle Scholar
  19. Kaplan LA, Pesce AJ (1984) Clinical chemistry: theory, analysis and correlation. Mosby, St. Louis, pp 1032–1036Google Scholar
  20. Krogdahl Å, Sundby A, Olli JJ (2004) Atlantic salmon (Salmon salar) and rainbow trout (Oncorhynchus mykiss) digest and metabolize nutrients differently. Effects of water salinity and dietary starch levels. Aquaculture 229:335–360CrossRefGoogle Scholar
  21. Kujala TS, Vienola MS, Klika KD, Loponen JM, Pihlaja K (2002) Betalain and phenolic composition of four beetroot (Beta vulgaris) cultivars. Eur Food Res Technol 214:505–510CrossRefGoogle Scholar
  22. Kumar N, Jadhao SB, Chandan NK, Kumar K, Jha AK, Bhushan S, Kumar S, Rana RS (2012) Dietary choline, betaine and lecithin mitigate endosulfan-induced stress in Labeo rohita fingerlings. Fish Physiol Biochem 38:989–1000CrossRefPubMedGoogle Scholar
  23. Leveelahti L, Rytkönen KT, Renshaw GMC, Nikinmaa M (2014) Revisiting redox-active antioxidant defences in response to hypoxic challenge in both hypoxia-tolerant and hypoxia-sensitive fish species. Fish Physiol Biochem 40:183–191CrossRefPubMedGoogle Scholar
  24. Lushchak VI, Bagnyukova TV, Lushchak OV, Storey JM, Storey KB (2005) Hypoxia and recovery perturb free radical processes and antioxidant potential in common carp (Cyprinus carpio) tissues. Int J Bichem Cell Biol 37:1319–1330CrossRefGoogle Scholar
  25. Lushchak VI, Bagnyukova TV (2006) Temperature increase results in oxidative stress in goldfish tissues. 1. Indices of oxidative stress. Comp Biochem Physiol C 143:30–35Google Scholar
  26. Madaro A, Olsen RE, Kristiansen TS, Ebbeson LOE, Nilsen TO, Flik G, Gorissen M (2015) Stress in Atlantic salmon: response to unpredictable chronic stress. J Exp Biol 218:2538–2550CrossRefPubMedGoogle Scholar
  27. Ming J, Xie J, Xu P, Ge X, Liu W, Ye J (2012) Effects of emodin and vitamin C on growth performance, biochemical parameters and two HSP70s mRNA expression of Wuchang bream (Megalobrama amblycephala Yih) under high temperature stress. Fish Shellfish Immunol 32:651–661CrossRefPubMedGoogle Scholar
  28. Mommsen TP, Vijayan MM, Moon TW (1999) Cortisol in teleosts: dynamics, mechanisms of action, and metabolic regulation. Rev Fish Biol Fish 9:211–268CrossRefGoogle Scholar
  29. Montero D, Tort L, Robaina L, Vergara JM, Izquierdo MS (2001) Low vitamin E in diet reduces stress resistance of gilthead seabream (Sparus aurta) juveniles. Fish Shellfish Immunol 11:473–490CrossRefPubMedGoogle Scholar
  30. Ortuño J, Esteban MA, Meseguer J (2003) Effect of dietary intake of vitamins C and E on the stress response of gilthead seabream (Sparus aurata L.) Fish Shellfish Immunol 14:145–156CrossRefPubMedGoogle Scholar
  31. Øverli Ø, Sørensen C, Kiessling A, Pottinger TG, Gjøen HM (2006) Selection for improved stress tolerance in rainbow trout (Oncorhynchus Mykiss) leads to reduced feed waste. Aquaculture 261:776–781CrossRefGoogle Scholar
  32. Pérez-Jiménez A, Peres H, Rubio VC, Oliva-Teles A (2012) The effect of hypoxia on intermediary metabolism and oxidative status in gilthead sea bream (Sparus aurata) fed on diets supplemented with methionine and white tea. Comp Biochem Physiol C 155:506–516Google Scholar
  33. Pichavant K, Maxime V, Thébault MT, Ollivier H, Garnier JP, Bousquet B, Diouris M, Boeuf G, Nonnotte G (2002) Effects of hypoxia and subsequent recovery on turbot (Scophthalmus maximus): hormonal changes and anaerobic metabolism. Mar Ecol Prog Ser 225:275–285CrossRefGoogle Scholar
  34. Pinedo-Gil J, Tomás-Vidal A, Larrán-García AM, Tomás-Almenar C, Jover-Cerdá M, Sanz-Calvo MA, Martín-Diana AB (2017a) Enhancement of quality of rainbow trout (Oncorhynchus mykiss) flesh incorporating barley on diet without negative effect on rearing parameters. Aquacult Int 25:1005–1023. CrossRefGoogle Scholar
  35. Pinedo-Gil J, Tomás-Vidal A, Jover-Cerdá M, Tomás-Almenar C, Sanz-Calvo MA, Martín-Diana AB (2017b) Red beet and betaine as ingredients in diets of rainbow trout (Oncorhynchus mykiss): effects on growth performance, nutrient retention and flesh quality. Arch Anim Nutr 71:486–505. CrossRefPubMedGoogle Scholar
  36. Rabeh NM (2015) Effect of red beetroot (Beta vulgaris L.) and its fresh juice against carbon tetrachloride induced hepatotoxicity in rats. World Appl Sci J 33(6):931–938Google Scholar
  37. Rollo A, Sulpizio R, Nardi M, Silvi S, Orpianesi C, Caggiano M, Cresci A, Carnevalli O (2006) Live microbial feed supplement in aquaculture for improvement of stress tolerance. Fish Physiol Biochem 32:167–177CrossRefGoogle Scholar
  38. Sadoul B, Leguen I, Colson V, Friggens NC, Prunet P (2015) A multivariate analysis using physiology and behaviour to characterize robustness in two isogenic lines of rainbow trout exposed to a confinement stress. Physiol Behav 140:139–147CrossRefPubMedGoogle Scholar
  39. Tan Q, Xie S, Zhu X, Lei W, Yang Y (2006) Effect of dietary carbohydrates sources on growth performance and utilization for gibel carp (Carassius auratus) and Chinese longsnout catfish (Leiocassis Longirostris Günther). Aquac Nutr 12:61–70CrossRefGoogle Scholar
  40. Tintos A, Míguez JM, Mancera JM, Soengas JL (2006) Development of a microtitre plate indirect ELISA for measuring cortisol in teleosts, and evaluation of stress responses in rainbow trout and gilthead sea bream. J Fish Biol 68:251–263CrossRefGoogle Scholar
  41. Van Anholt RD, Spanings FAT, Koven WM, Nixon O, Wendelaar Bonga SE (2004) Arachidonic acid reduces the stress response of gilthead seabream, Sparus aurata L. J Exp Biol 207:3419–3430CrossRefPubMedGoogle Scholar
  42. Virtanen E (1995) Piecing together the betaine puzzle. Feed Min 3:12–17Google Scholar
  43. Wu XY, Liu YJ, TIan LX, Mai KS, Yang HJ (2007) Utilization of several different carbohydrate sources by juvenile yellowfin seabream (Sparus latus). J Fish China 31(4):463–471Google Scholar
  44. Yoshida Y, Itoh N, Hayakawa M, Piga R, Cynshi O, Jishage K, Niki E (2005) Lipid peroxidation induced by carbon tetrachloride and its inhibition by antioxidant as evaluated by an oxidative stress marker, HODE. Toxicol Appl Pharmacol 208:87–97CrossRefPubMedGoogle Scholar
  45. Zeng L, Wang YH, Ai CX, Zheng JL, Wu CW, Cai R (2016) Effects of β-glucan on ROS production and energy metabolism in yellow croaker (Pseudosciaena crocea) under acute hypoxic stress. Fish Physiol Biochem 42:1395–1405CrossRefPubMedGoogle Scholar
  46. Zolderdo AJ, Algera DA, Lawrence MJ, Gilmour KM, Fast MD, Thuswaldner J, Willmore WG, Cooke SJ (2016) Stress, nutrition and parental care in a teleost fish: exploring mechanisms with supplemental feeding and cortisol manipulation. J Exp Biol 219:1237–1248CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Julia Pinedo-Gil
    • 1
  • Ana Belén Martín-Diana
    • 2
  • Daniela Bertotto
    • 3
  • Miguel Ángel Sanz-Calvo
    • 2
  • Miguel Jover-Cerdá
    • 1
  • Ana Tomás-Vidal
    • 1
  1. 1.Research Group of Aquaculture and Biodiversity, Institute of Animal Science and TechnologyUniversitat Poliècnica de ValènciaValenciaSpain
  2. 2.Subdirection of Research and Technology, Agro-Technological Institute of Castilla y León, Consejería de Agricultura y GanaderíaValladolidSpain
  3. 3.Department of Comparative Biomedicine and Food ScienceUniversity of PadovaLegnaroItaly

Personalised recommendations