Fish Physiology and Biochemistry

, Volume 43, Issue 2, pp 591–602 | Cite as

Effects of amino acid supplementations on metabolic and physiological parameters in Atlantic cod (Gadus morhua) under stress

  • Marcelino Herrera
  • María Antonia Herves
  • Inmaculada Giráldez
  • Kristin Skar
  • Hanne Mogren
  • Atle Mortensen
  • Velmurugu Puvanendran


The effects of tryptophan (Trp) and phenylalanine (Phe) diet supplementation on the stress and metabolism of the Atlantic cod have been studied. Fish were fed diet supplemented with Trp or Phe or control diet for 1 week. At the end of the feeding trial, fish were subjected to air exposure or heat shock. Following samples of blood, liver and muscle were taken from the fish and were analyzed for stress and metabolic indicators. After an air exposure, plasma cortisol levels in fish fed with Trp and Phe diets were lower compared to the fish fed the control diet. Diets containing both amino acids increased significantly the liver transaminase activities in juvenile cod. During thermal stress, high Trp contents had significant effects on fructose biphosphatase activity though Phe did not. Overall, activities of glucose 6-phosphate dehydrogenase, pyruvate kinase, and phosphofructokinase increased significantly for both amino acid diets. For the thermal stress, fish had the highest values of those activities for the 3Trp diet. Trp content in the diet had significant effects on the transaminase activity in muscle during air stress compared to fish fed control and Phe diets. Muscle alanine transaminase activity for thermal stress in fish fed any diet was not significantly different from the control. Both Trp and Phe supplementations reduced the stress markers in the cod; hence, they could be used as additives for the stress attenuation. However, they also raised the activity of key enzymes in glycolysis and gluconeogenesis, mainly the Trp diets.


Stress Cod Welfare Tryptophan Phenylalanine Metabolism 



We would like to thank all the staff at the CMAR in Tromsø for their help in successfully conducting this experiment. This work has been supported by Aquaexcel project (FP7). M. Herrera post-doc contract is supported by INIA-FSE.

Compliance with ethical standards

The experiment complied with the guidelines of the European Union Council (2010/63/EU) and the Norwegian Animal Welfare Act for the use of laboratory animals. According to the Spanish RD1201/2005, M. Herrera is certified (type C) for working and designing experiments with animals.


  1. Ahmed I (2009) Dietary total aromatic amino acid requirement and tyrosine replacement value for phenylalanine in Indian major carp: Cirrhinus mrigala (Hamilton) fingerlings. J Appl Ichthyol 25:719–727CrossRefGoogle Scholar
  2. Ashley PJ (2007) Fish welfare: current issues in aquaculture. Appl Anim Behav Sci 104(3–4):199–235CrossRefGoogle Scholar
  3. Banderet LE, Lieberman HR (1989) Treatment with tyrosine, a neurotransmitter precursor, reduces environmental stress in human. Brain Res Bull 22:759–762CrossRefPubMedGoogle Scholar
  4. Barton BA, Iwama GK (1991) Physiological changes in fish from stress in aquaculture with emphasis on the response and effects of corticosteroids. Ann Rev Fish Dis 129:3–26CrossRefGoogle Scholar
  5. Barton BA (2002) Stress in fishes: a diversity of responses with particular reference to changes in circulating corticosteroids. Integr Comp Biol 42:517–525CrossRefPubMedGoogle Scholar
  6. Barton BA, Morgan JD, Vijayan MM (2002) Physiological and condition-related indicators of environmental stress on fish. In: Adams SM (ed) Biological indicators of aquatic ecosystems stress. American Fisheries Society, BethesdaGoogle Scholar
  7. Basic D, Schjolden J, Krogdahl A, Krogh KV, Hillestad M, Winberg S, Mayer I, Skjerve E, Höglund E (2013) Changes in regional brain monoaminergic activity and temporary down-regulation in stress response from dietary supplementation with L-tryptophan in Atlantic cod (Gadus morhua). Brit J Nutr 109:2166–2174CrossRefPubMedGoogle Scholar
  8. Borlongan IG (1992) Dietary requirement of milkfish (Chanos chanos Forsskal) for total aromatic amino acids. Aquaculture 102:309–317CrossRefGoogle Scholar
  9. Brady K, Brown JW, Thurmond JB (1980) Behavioral and neurochemical effects of dietary tyrosine in young and aged mice following cold-swim stress. Pharmacol Biochem Be 12:667–674CrossRefGoogle Scholar
  10. Costas B, Conceição LEC, Dias J, Novoa B, Figueras A, Afonso A (2011) Dietary arginine and repeated handling increase disease resistance and modulate innate immune mechanisms of Senegalese sole (Solea senegalensis Kaup, 1858). Fish Shellfish Immun 3:838–847CrossRefGoogle Scholar
  11. Cotoia A, Scrima R, Gefter JV, Piccoli C, Cinnella G, Dambrosio M, Fink MP, Capitanio N (2014) P-hydroxyphenylpyruvate, an intermediate of the Phe/Tyr catabolism, improves mitochondrial oxidative metabolism under stressing conditions and prolongs survival in rats subjected to profound hemorrhagic shock. PLoS One 9(3):e90917CrossRefPubMedPubMedCentralGoogle Scholar
  12. Dai Z, Wu Z, Ji S, Wu G (2014) Analysis of amino acid composition in proteins of animal tissues and foods as pre-column o-phthaldialdehyde derivatives by HPLC with fluorescence detection. J Chromatogr B 964:116–127CrossRefGoogle Scholar
  13. Hansen ØJ, Puvanendran V (2010) Fertilization success and blastomere morphology as predictors of egg and juvenile quality for domesticated Atlantic cod, Gadus morhua, broodstock. Aquac Res 41:1791–1798CrossRefGoogle Scholar
  14. Hansen ØJ, Puvanendran V, Bangera R (2015) Do maternal age and experience contribute to better growth, survival and disease resistance of offspring in Atlantic cod (Gadus morhua)? Aquacult Int 23:1157–1164CrossRefGoogle Scholar
  15. Hansen ØJ, Puvanendran V, Mortensen A (2013) Importance of broodstock holding temperature on fecundity and egg quality in three groups of photo-manipulated Atlantic cod broodstock. Aquac Res 44:140–150CrossRefGoogle Scholar
  16. Herrera M, Aragão A, Hachero I, Ruiz-Jarabo I, Vargas-Chacoff L, Mancera JM, Conceição L (2012) Physiological short-term response to sudden salinity change in the Senegalese sole (Solea senegalensis). Fish Physiol Biochem 38:1741–1751CrossRefPubMedGoogle Scholar
  17. Herrera M, Ruiz-Jarabo I, Vargas-Chacoff L, De La Roca E, Mancera JM (2015) Metabolic enzyme activities in relation to crowding stress in the wedge sole (Dicologoglossa cuneata). Aquac Res 46:2808–2818CrossRefGoogle Scholar
  18. Höglund E, Bakke MJ, Øverli Ø, Winberg S, Nilsson GE (2005) Suppression of aggressive behaviour in juvenile Atlantic cod (Gadus morhua) by l-tryptophan supplementation. Aquaculture 249:525–531CrossRefGoogle Scholar
  19. Höglund E, Balm PHM, Winberg S (2000) Skin darkening, a potential social signal in subordinate Arctic charr (Salvelinus alpinus): the regulatory role of brain monoamines and pro-opiomelanocortin derived peptides. J Exp Biol 203:1711–1721PubMedGoogle Scholar
  20. Höglund E, Kolm N, Winberg S (2001) Stress-induced effects on brain serotonergic activity, plasma cortisol and aggressive behaviour in Arctic charr (Salvelinus alpinus) is counteracted by L-dopa. Physiol Behav 74:381–389CrossRefPubMedGoogle Scholar
  21. Hoseini SM, Hosseini SA, Soudagar M (2012) Dietary tryptophan changes serum stress markers, enzyme activity, and ions concentration of wild common carp Cyprinus carpio exposed to ambient copper. Fish Physiol Biochem 38(5):1419–1426CrossRefPubMedGoogle Scholar
  22. Hseu J, Lu F, Su H, Wang L, Tsai C, Hwang P (2003) Effect of exogenous tryptophan on cannibalism, survival and growth in juvenile grouper, Epinephelus coioides. Aquaculture 218(1–4):251–263CrossRefGoogle Scholar
  23. Iwama GK, Afonso LOB, Vijayan MM (2006) Stress in fishes. In: Evans DH, Claiborne JB (eds) The physiology of fishes. CRC Press, New York, pp. 319–342Google Scholar
  24. Laiz-Carrión R, Martín del Río MP, Mínguez J, Mancera JM, Soengas JL (2003) Influence of cortisol on osmoregulation and energy metabolism in gilthead seabream Sparus aurata. J Exp Zool 298A:105–118CrossRefGoogle Scholar
  25. Laranja LQJ Jr, Quinitio EM, Catacutan MR, Coloso RM (2010) Effects of dietary L-tryptophan on the agonistic behavior, growth and survival of juvenile mud crab Scylla serrata. Aquaculture 310:84–90CrossRefGoogle Scholar
  26. Le Floc’h N, Otten W, Merlot E (2011) Tryptophan metabolism, from nutrition to potential therapeutic applications. Amino Acids 41:1195–1205CrossRefPubMedGoogle Scholar
  27. Lehnert H, Reinstein DK, Strowbridge BW, Wurtman RJ (1984) Neurochemical and behavioral consequences of acute and uncontrollable stress: effects of dietary tyrosine. Brain Res 303:215–223CrossRefPubMedGoogle Scholar
  28. Lepage O, Tottmar O, Winberg S (2002) Elevated dietary intake of L-tryptophan counteracts the stress-induced elevation of plasma cortisol in rainbow trout (Oncorhynchus mykiss). J Exp Biol 205:3679–3687PubMedGoogle Scholar
  29. Lepage O, Vílchez IM, Pottinger TG, Winberg S (2003) Time-course of the effect of dietary L-tryptophan on plasma cortisol levels in rainbow trout Oncorhynchus mykiss. J Exp Biol 206:3589–3599CrossRefPubMedGoogle Scholar
  30. Li P, Mai K, Trushenski J, Wu G (2009) New developments in fish amino acid nutrition: towards functional and environmentally oriented aquafeeds. Amino Acids 37(1):43–53CrossRefPubMedGoogle Scholar
  31. Maricchiolo G, Mirto S, Caruso G, Caruso T, Bonaventura R, Celi M, Matranga V, Genovese L (2011) Welfare status of cage farmed European sea bass (Dicentrarchus labrax): a comparison between submerged and surface cages. Aquaculture 314(1–4):173–181CrossRefGoogle Scholar
  32. Martin DW, Mayers PA, Rodwell VW (1983) Harper’s review of biochemistry. Lange Medical Publications, Maruzen, AsiaGoogle Scholar
  33. Martins CIM, Silva PIM, Costas B, Larsen BK, Santos GA, Conceição LEC, Dias J, Øverli Ø, Höglund E, Schrama JW (2013) The effect of tryptophan supplemented diets on brain serotonergic activity and plasma cortisol under undisturbed and stressed conditions in grouped-housed Nile tilapia Oreochromis niloticus. Aquaculture 400-401:129–134CrossRefGoogle Scholar
  34. Mommsen TP, Vijayan MM, Moon TW (1999) Cortisol in teleosts: dynamics, mechanisms of action, and metabolic regulation. Rev Fish Biol Fisher 9(3):211–268CrossRefGoogle Scholar
  35. Montero D, Izquierdo MS, Tort L, Robaina L, Vergara JM (1999) High stocking density produces crowding stress altering some physiological and biochemical parameters in gilthead seabream, Sparus aurata, juveniles. Fish Physiol Biochem 20:53–60CrossRefGoogle Scholar
  36. NRC (National Research Council) (2011) Nutrient requirements of fish and shrimp. National Academy Press, Washington (DC 978-0-309-16338-5 (376 + XVI pp.))Google Scholar
  37. Øverli Ø, Harris CA, Winberg S (1999) Short term effects of fights for social dominance and the establishment of dominant-subordinate relationships on brain monoamines and cortisol in rainbow trout. Brain Behav Evolut 54:263–275CrossRefGoogle Scholar
  38. Papoutsoglou SE, Karakatsouli N, Chiras G (2005) Dietary l-tryptophan and tank colour effects on growth performance of rainbow trout (Oncorhynchus mykiss) juveniles reared in a recirculating water system. Aquac Eng 32:277–284CrossRefGoogle Scholar
  39. Phillips RS, Miles EW, Cohen LA (1984) Interactions of tryptophan synthase, tryptophanase, and pyridoxal phosphate with oxindolyl-L-alanine and 2, 3-dihydro-L-tryptophan: support for an indolenine intermediate in tryptophan metabolism. Biochemistry 23(25):6228–6234CrossRefPubMedGoogle Scholar
  40. Polakof S, Arjona FJ, Sangiao-Alvarellos S, Martín del Río MP, Mancera JM, Soengas JL (2006) Food deprivation alters osmoregulatory and metabolic responses to salinity acclimation in gilthead sea bream Sparus auratus. J Comp Physiol 176B:441–452CrossRefGoogle Scholar
  41. Ren M, Liua B, Habte-Tsion H-M, Ge X, Xie J, Zhoua Q, Liang H, Zhao Z, Pana L (2015) Dietary phenylalanine requirement and tyrosine replacement value for phenylalanine of juvenile blunt snout bream, Megalobrama amblycephala. Aquaculture 442:51–57CrossRefGoogle Scholar
  42. Saavedra M, Conceição LEC, Barr Y, Helland S, Pousao-Ferreira P, Yúfera M, Dinis MT (2010) Tyrosine and phenylalanine supplementation on Diplodus sargus larvae: effect on growth and quality. Aquac Res 41:1523–1532CrossRefGoogle Scholar
  43. Saha N, Jyrwa LM, Das M, Biswas K (2011) Influence of increased environmental water salinity on gluconeogenesis in the air-breathing walking catfish, Clarias batrachus. Fish Physiol Biochem 37:681–692CrossRefPubMedGoogle Scholar
  44. Sangiao-Alvarellos S, Guzmán JM, Laiz-Carrión R, Míguez JM, Martín del Rio MP, Mancera JM, Soengas JL (2005) Interactive effects of high stocking density and food deprivation on carbohydrate metabolism in several tissues of gilthead sea bream (Sparus aurata L.). J Exp Zool 303A:761–775CrossRefGoogle Scholar
  45. Sangiao-Alvarellos S, Laíz-Carrión R, Guzmán JM, Martín del Río MP, Migues JM, Mancera JM, Soengas JL (2003) Acclimation of S. auratus to various salinities alters energy metabolism of osmoregulatory and non-osmoregulatory organs. Am J Phys 285: R897–R907.Google Scholar
  46. Santos GA, Schrama JW, Mamauag REP, Rombout JHWM, Verreth JAJ (2010) Chronic stress impairs performance, energy metabolism and welfare indicators in European seabass (Dicentrarchus labrax): the combined effects of fish crowding and water quality deterioration. Aquaculture 299:73–80CrossRefGoogle Scholar
  47. Sawin EA, Murali SG, Ney DM (2014) Differential effects of low-phenylalanine protein sources on brain neurotransmitters and behavior in C57Bl/6-Pah enu2 mice. Mol Genet Metab 111:452–461CrossRefPubMedPubMedCentralGoogle Scholar
  48. Shafik M, Ibrahime H, Elyazeid IA, Abass O, Saad HM (2014) The stress of phenylalanine on rats to study the phenylketonuria at biochemical and molecular level. J App Pharmac Sci 4(04):024–029Google Scholar
  49. Tejpal CS, Pal AK, Sahu NP, Ashish Kumar J, Muthappa NA, Vidya S, Rajan MG (2009) Dietary supplementation of L-tryptophan mitigates crowding stress and augments the growth in Cirrhinus mrigala fingerlings. Aquaculture 293(3–4):272–277CrossRefGoogle Scholar
  50. Tiwari S, Singh A (2006) Biochemical stress response in freshwater fish Channa punctatus induced by aqueous extracts of Euphorbia tirucalli plant. Chemosphere 64:36–42CrossRefPubMedGoogle Scholar
  51. Toni C, Martos-Sitcha JA, Ruiz-Jarabo I, Mancera JM, Martínez-Rodríguez G, Pinheiro CG, Heinzmann BM, Baldisserotto B (2015) Stress response in silver catfish (Rhamdia quelen) exposed to the essential oil of Hesperozygis ringens. Fish Physiol Biochem 41:129–138CrossRefPubMedGoogle Scholar
  52. Vargas-Chacoff L, Arjona FJ, Ruiz-Jarabo I, Páscoa I, Gonçalves O, Martín del Río MP, Mancera JM (2009) Seasonal variation in osmoregulatory and metabolic parameters in earthen pond-cultured gilthead sea bream Sparus auratus. Aquac Res 40:1279–1290CrossRefGoogle Scholar
  53. Vijayan MM, Ballantyne JS, Leatherland JF (1990) High stocking density alters the energy metabolism of brook charr, Salvelinus fontinalis. Aquaculture 88:371–381CrossRefGoogle Scholar
  54. Wendelaar Bonga SE (1997) The stress response in fish. Physiol Revi 7:591–265Google Scholar
  55. Winberg S, Nilsson GE (1992) Induction of social dominance by L-dopa treatment in Arctic charr. Neuroreport 3:243–246CrossRefPubMedGoogle Scholar
  56. Yao K, Fang J, Yin Y-L, Feng Z-M, Tang Z-R, Wu G (2011) Tryptophan metabolism in animals: important roles in nutrition and health. Front Biosci S3:286–297CrossRefGoogle Scholar
  57. Zehra S, Khan MA (2014) Dietary phenylalanine requirement and tyrosine replacement value for phenylalanine for fingerling Catla catla (Hamilton). Aquaculture 433:256–265CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.IFAPA Centro Agua del PinoCartayaSpain
  2. 2.Facultad de Ciencias ExperimentalesUniversidad de HuelvaHuelvaSpain
  3. 3.Centre for Marine Aquaculture ResearchKvaløyaNorway
  4. 4.NofimaTromsøNorway

Personalised recommendations