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Effects of dietary DHA and α-tocopherol on bone development, early mineralisation and oxidative stress in Sparus aurata (Linnaeus, 1758) larvae

  • M. S. Izquierdo (a1), M. Scolamacchia (a1), M. Betancor (a1), J. Roo (a1), M. J. Caballero (a1), G. Terova (a2) and P. E. Witten (a3)...

DHA deficiency has been related to skeletal malformations in fish, but high DHA levels have produced controversial results that could relate to the oxidative status of fish tissues in the different reports. In the present study, gilthead seabream (Sparus aurata) larvae were fed deficient, adequate or high DHA levels, or high DHA levels supplemented with the antioxidant α-tocopherol. Larvae fed deficient DHA levels tended to be smaller, and showed the highest incidence of urinary bladder calculi, lordosis and kyphosis and the lowest number of mineralised vertebrae for any given size class. Elevation of dietary DHA increased larval growth and significantly enhanced the expression of the insulin-like growth factor 1 (IGF-1) gene. However, a DHA level increase up to 5 % raised the degree of lipid oxidation in larval tissues and deformities in cranial endochondral bones and in axial skeletal haemal and neural arches. The increase in dietary α-tocopherol supplementation in high-DHA feeds reduced again the occurrence of skeletal deformities. Moreover, the expression of genes coding for specific antioxidants such as catalase, superoxide dismutase or glutathione peroxidase, which neutralised reactive oxygen substances formed by increased dietary DHA, was significantly decreased in larvae fed high α-tocopherol levels. These results denoted the importance of DHA for early bone formation and mineralisation. Low dietary DHA levels delay early mineralisation and increase the risk of cranial and axial skeletal deformities. Excessive DHA levels, without an adequate balance of antioxidant nutrients, increase the production of free radicals damaging cartilaginous structures before bone formation.

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      Effects of dietary DHA and α-tocopherol on bone development, early mineralisation and oxidative stress in Sparus aurata (Linnaeus, 1758) larvae
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*Professor M. S. Izquierdo, fax +34 928132908, email;
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1Lewis-McCrea, M & Lall, SP (2007) Effects of moderately oxidized dietary lipid and the role of vitamin E on the development of skeletal abnormalities in juvenile Atlantic halibut (Hippoglossus hippoglossus). Aquaculture 262, 142155.
2Satoh, S, Poe, WE & Wilson, RP (1989) Effect of supplementation of phytate and/or 3-calcium phosphate on wet gain, feed efficiency and zinc content in vertebrae of channel catfish. Aquaculture 80, 155161.
3Takeuchi, T (1999) Mechanism and prevention of color abnormality in flounder. Aquanet 2, 4852.
4Takeuchi, T, Dedi, J, Haga, Y, et al. (1998) Effect of vitamin A compounds on bone deformity in larval Japanese flounder Paralichthys olivaceus. Aquaculture 169, 155165.
5Cahu, CL, Zambonino Infante, JL & Barbosa, V (2003) Phospholipid level in dietary lipid fraction is determining for sea bass (Dicentrarchus labrax) larval development. Br J Nutr 90, 2128.
6Izquierdo MS (1988) Essential fatty acid requirements of marine fish larvae: modification of live prey lipid composition. PhD Thesis, La Laguna University, Spain, 205 pp.
7Roo FJ (2009) Improvement in the production technology of red porgy (Pagrus pagrus) larvae and fry: importance of rearing conditions and diet nutritional value on their quality. PhD Thesis, Universidad de Las Palmas de Gran Canaria, Spain.
8Lewis-McCrea, LM & Lall, SP (2010) Effects of phosphorus and vitamin C deficiency, vitamin A toxicity, and lipid peroxidation on skeletal abnormalities in Atlantic halibut (Hippoglossus hippoglossus). J App Ichthyol 26, 334343.
9Kanazawa, A, Teshima, S, Inamori, S, et al. (1981) Effects of phospholipids on growth, survival rate and incidence of malformation in the larval ayu. Mem Fac Fish Kagoshima Univ 30, 301309.
10Izquierdo, MS, Socorro, J & Roo, J (2010) Studies on the appearance of skeletal anomalies in red porgy: effect of culture intensiveness, feeding habits and nutritional quality of live preys. J App Ichthyol 26, 320326.
11Gapasin, R & Duray, M (2001) Effects of DHA-enriched live food on growth, survival and incidence of opercular deformities in milkfish (Chanos chanos). Aquaculture 193, 4963.
12Villeneuve, L, Gisbert, E, Zambonino Infante, JL, et al. (2005) Effect of nature of dietary lipids on European sea bass morphogenesis: implication of retinoid receptors. Br J Nutr 94, 877884.
13Betancor, M, Caballero, MJ, Terova, G, et al. (2012) Selenium inclusion decreases oxidative stress indicators and muscle injuries in sea bass larvae fed high DHA microdiets. Br J Nutr 108, 21152128.
14Zhang, X, Xie, P, Li, D, et al. (2009) Time-dependent oxidative stress responses of crucian carp (Carassius auratus) to intraperitoneal injection of extracted microcystins. Bull Environ Contam Toxicol 82, 574578.
15Pi, J, Zhang, Q, Fu, JG, et al. (2010) ROS signaling, oxidative stress and Nrf2 in pancreatic beta-cell function. Toxicol Appl Pharmacol 244, 7783.
16Jin, Y, Zheng, S, Pu, Y, et al. (2010) Cypermethrin has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in adult zebrafish (Danio rerio). Chemosphere 82, 398404.
17Carnevalli, O, de Vivo, L, Sulpicio, R, et al. (2006) Growth improvement by probiotic in European sea bass (Dicentrarchus labrax, L.) juveniles, with particular attention to IGF-1, myostatin and cortisol gene expression. Aquaculture 258, 430438.
18Watkins, BA & Seifert, MF (2000) Conjugated linoleic acid and bone biology. J Am Coll Nutr 19, 478486.
19Lall, SP & Lewis-McCrea, LM (2007) Role of nutrients in skeletal metabolism and pathology in fish – an overview. Aquaculture 267, 319.
20Hall, BK & Witten, PE (2007) Plasticity of and transitions between skeletal tissues in vertebrate evolution and development. In Major Transitions in Vertebrate Evolution, pp. 5669 [Anderson, JS and Sues, DH, editors]. Bloomington, IN: Indiana University Press.
21Huxley, TH (1859) Observations on the development of some parts of the skeleton of fishes. Quar J Microscop Sci 7, 3346.
22Arratia, G, Schultze, HP & Casciotta, J (2001) Vertebral column and associated elements in dipnoans and comparison with other fishes: development and homology. J Morphol 250, 101172.
23Fernández-Palacios, H, Izquierdo, MS, Robaina, L, et al. (1995) Effect of n-3 HUFA level in broodstock diets on egg quality of gilthead seabream (Sparus aurata L.). Aquaculture 132, 325337.
24Izquierdo, MS, Watanabe, T, Takeuchi, T, et al. (1989) Optimal EFA levels in Artemia to meet the EFA requirements of red seabream Pagrus major. In The Current Status of Fish Nutrition in Aquaculture, pp. 221232 [Takeda, M and Watanabe, T, editors]. Tokyo: Japan Translation Centre.
25Vandewalle, P, Gluckmann, I & Wagemans, F (1998) A critical assessment of the alcian blue/alizarin double staining in fish larvae and fry. Belgian J Zool 128, 9395.
26Boglione, C, Gagliardi, F, Scardi, M, et al. (2001) Skeletal descriptors and quality assessment in larvae and post-larvae of wild caught and hatchery reared gilthead sea bream (Sparus aurata L. 1758). Aquaculture 192, 122.
27Folch, J, Lees, M & Sloane Stanley, GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226, 497509.
28Christie, WW (1989) Gas Chromatography and Lipids: A Practical Guide. Glasgow: The Oily Press.
29Izquierdo, MS, Arakawa, T, Takeuchi, T, et al. (1992) Effect of n-3 HUFA levels in Artemia on growth of larval Japanese flounder (Paralichthys olivaceus). Aquaculture 105, 7382.
30AOAC (1995) Official Methods of Analysis of the Association Analytical Chemistries. Arlington, VA: AOAC.
31Burk, RF, Trumble, MJ & Lawrence, RA (1980) Rat hepatic cytosolic GSH-dependent enzyme protection against lipid peroxidation in the NADPH microsomal lipid peroxidation system. Biochim Biophys Acta 618, 3541.
32Tocher, DR, Mourente, G, van der Eecken, A, et al. (2002) Effects of dietary vitamin E on antioxidant defence mechanisms of juvenile turbot (Scophtalmus maximus), halibut (Hippoglossus hippoglossus L.) and sea bream (Sparus aurata). Aquaculture Nutr 8, 195207.
33Izquierdo, MS & Koven, W (2010) Lipids. In Larval Fish Nutrition, pp. 4782 [Holt, J, editor]. New York: Wiley-Blackwell.
34Izquierdo, MS (2005) Essential fatty acids requirements in Mediterranean fish species. Cah Opt Med 63, 91101.
35Liu, J, Caballero, MJ, Izquierdo, MS, et al. (2002) Necessity of dietary lecithin and eicosapentaenoic acid for growth, survival, stress resistance and lipoprotein formation in gilthead seabream Sparus aurata. Fish Sci 225, 325340.
36Rodríguez, C, Pérez, JA, Badía, P, et al. (1998) The n-3 highly unsaturated fatty acids requirements of gilthead sea bream (Sparus aurata L.) larvae when using an appropriate DHA/EPA ratio in the diet. Aquaculture 169, 923.
37Kranenbarg, S, Waarsing, JH, Muller, M, et al. (2005) Lordotic vertebrae in sea bass (Dicentrarchus labrax L.) are adapted to increased loads. J. Biomech 38, 1239.
38Kruger, MC, Coetzee, M, Haag, M, et al. (2010) Long-chain polyunsaturated fatty acids: selected mechanisms of action on bone. Prog Lipid Res 49, 438449.
39Kruger, MC & Schollum, LM (2005) Is docosahexaenoic acid more effective than eicosapentaenoic acid for increasing calcium bioavailability? Prostaglandins Leukot Essen Fatty Acids 73, 327334.
40Raisz, LG, Alander, CB & Simmons, HA (1989) Effects of prostaglandin E3 and eicosapentaenoic acid on rat bone in organ culture. Prostaglandins 37, 615625.
41Serhan, CN (2005) Novel eicosanoid and docosanoid mediators: resolvins, docosatrienes and neuroprotectins. Curr Opin Clin Nutr Metab Care 8, 115121.
42Ganga, R, Tort, L, Acerete, L, et al. (2006) Modulation of ACTH-induced cortisol release by polyunsaturated fatty acids in interregnal cells from gilthead seabream, Sparus aurata. J Endocrinol 190, 3945.
43Ganga, R, Bell, GJ, Montero, D, et al. (2011) Adrenocorticotrophic hormone-stimulated cortisol release by the head kidney interrenal tissue from sea bream (Sparus aurata) fed with linseed oil and soyabean oil. Br J Nutr 105, 238247.
44Fernandes, G, Bhattacharaya, A, Rahman, M, et al. (2008) Effects of n-3 fatty acids on autoimmunity and osteoporosis. Front Biosci 13, 40154020.
45Pozios, KC, Ding, J, Degger, B, et al. (2001) IGFs stimulate zebrafish cell proliferation by activating MAP kinase ans PI3-kinase-signalling pathways. Am J Physiol 280, 12301239.
46Collins, DA & Chambers, TJ (1991) Effect of prostaglandins E1, E2, and F2a on osteoclast formation in mouse bone marrow cultures. J Bone Miner Res 6, 157164.
47Pombinho, AR, Laize, V, Molha, DM, et al. (2004) Development of two bone-derived cell lines from the marine teleost Sparus aurata: evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin. Cell Tissue Res 315, 393406.
48Komondouros, G (2010) Morpho-anatomical abnormalities in Mediterranean marine aquaculture. Recent Adv Aquacult Res 2010, 125148.
49Betancor, MB, Atalah, E, Caballero, MJ, et al. (2010) Alpha-tocopherol in weaning diets for European sea bass (Dicentrarchus labrax) improves survival and reduces tissue damage caused by excess dietary DHA contents. Aquaculture Nutr 17, 112122.
50Beer, AM & Wegener, T (2011) Vitamin E for gonarthrosis and coxarthrosis: results of a postmarketing surveillance study. Fortschr Med 153, 1420.
51Dabrowski, K, El-Fiky, N, Köck, G, et al. (1990) Requirement and utilization of ascorbic acid and ascorbic sulfate in juvenile rainbow trout. Aquaculture 91, 317337.
52Atalah E (2008) Importance of the proportions of polyunsaturated fatty acids and antioxidants in larval development of marine fish. PhD Thesis, Universidad de Las Palmas de Gran Canaria, Spain.
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