Fish Physiology and Biochemistry

, Volume 14, Issue 2, pp 125–137 | Cite as

Effects of salinity on the fatty acid compositions of total lipid and individual glycerophospholipid classes of Atlantic salmon (Salmo salar) and turbot (Scophthalmus maximus) cells in culture

  • Douglas R. Tocher
  • John D. Castell
  • James R. Dick
  • John R. Sargent


Cells from a relatively stenohaline marine species, turbot (Scophthalmus maximus) (TF) and an anadromous species, Atlantic salmon (AS) were cultured in media supplemented with NaCl to produce OPs varying from 300 to 500 mOsm kg−1 and the direct effects of OP (salinity) on the fatty acid compositions of the main glycerophospholipid classes were determined. The most dramatic effects of salinity on total lipid fatty acids were observed in polyunsaturated fatty acids (PUFA) in TF cells. There was a graded decrease in the percentage of 18:2n-9, and consequently total n-9 PUFA, and concomitantly increased percentages of both total n-3 and n-6 PUFA with increasing salinity. The increased n-3 and n-6 PUFA was due to significantly increased percentages of the major fatty acids in each of these groups, namely 22:6n-3 and 20:4n-6, respectively. The reciprocal changes in n-9 PUFA and n-3/n-6 PUFA in TF cell total lipid resulted in the percentage of total PUFA not being significantly affected by changes in salinity. The graded decrease in 18:2n-9 with increasing salinity in TF cells was observed in all the major glycerophospholipids but especially PE, PI and PS. Increasing salinity resulted in graded increases in the percentages of 22:6n-3 in PE and PS in TF cells. The quantitatively greatest increase in the percentage of n-6 PUFA in TF cells occurred with 20:4n-6 in PC, PE and PL. There were less significant changes in the fatty acid compositions of glycerophospholipids in AS cells. However, the proportion of total n-3 + n-6 PUFA in PE varied reciprocally with the proportion of dimethylacetals in response to salinity. Similar reciprocal changes between fatty acids in response to salinity were also evident in the quantitatively more minor glycerophospholipids PS and Pl. In PS, the percentage of 22:6n-3 was significantly lower at 400 mOsm kg−1 whereas the proportion of total monoenes was significantly higher at that salinity. A similar inverse relationship between total monoenes and 20:4n-6 (and, to a lesser extent total saturates) in response to salinity was noted in PI. The results show that environmental salinity, without whole-body physiological stimuli, has direct effects on the fatty acid composition of major glycerophospholipid classes in fish cells and that these effects differ in cells from different fish species


Atlantic salmon turbot cell culture salinity fatty acids glycerophospholipids 



analysis of variance


butylated hydroxytoluene


bovine serum albumin




Eagle's minimal essential medium


fetal calf serum


gas chromatography


Hank's balanced salt solution (without Ca2+ and Mg2+)


osmotic pressure










polyunsaturated fatty acid


thin-layer chromatography


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References cited

  1. Al-Amoudi, M.M. 1987. Acclimation of commercially culturedOreochromis species to sea water — an experimental study. Aquaculture 65: 333–342.Google Scholar
  2. Alexis, M.N., Papaparaskeva-Papoutsoglou, E. and Papoutsoglou, S. 1984. Influence of acclimation temperature on the osmotic regulation and survival of rainbow trout (Salmo gardneri) rapidly transferred from fresh water to sea water. Aquaculture 40: 333–341.Google Scholar
  3. Arnold-Reed, D.E. and Balment, R.J. 1991. Salinity tolerance and its seasonal variation in the flounder,Platichthys flesus. Comp. Biochem. Physiol. 99A: 145–149.Google Scholar
  4. Assem, H. and Hanke, W. 1981. Cortisol and osmotic adjustment of the euryhaline teleost,Sarotherodon mossambicus. Gen. Comp. Endocrinol. 43: 370–380.Google Scholar
  5. Avella, M., Young, G., Prunet, P. and Schreck, C.B. 1990. Plasma prolactin and cortisol concentrations during salinity challenges of coho salmon (Oncorhynchus kisutch) at smolt and post-smolt stages. Aquaculture 91: 359–372.Google Scholar
  6. Bell, M.V., Henderson, R.J. and Sargent, J.R. 1985b. Changes in the fatty acid composition of phospholipids from turbot (Scophthalmus maximus) in relation to dietary polyunsaturated fatty acid deficiencies. Comp. Biochem. Physiol. 81B: 193–198.Google Scholar
  7. Bell, M.V., Simpson, C.M.F. and Sargent, J.R. 1983. (n-3) and (n-6) Polyunsaturated fatty acids in the phosphoglycerides of salt-secreting epithelia from two marine fish species. Lipids 18: 720–726.Google Scholar
  8. Bell, M.V., Henderson, R.J., Pirie, B.J.S. and Sargent, J.R. 1985a. Effects of dietary polyunsaturated fatty acid deficiencies on mortality, growth and gill structure in the turbot,Scophthalmus maximus. J. Fish Biol. 26: 181–191.Google Scholar
  9. Bentinck-Smith, J., Beleau, M.H., Waterstrat, P., Tucker, C.S., Stiles, F., Bowser, P.R. and Brown, L.A. 1987. Biochemical reference ranges for commercially reared channel catfish. Progr. Fish Cult. 49: 108–114.Google Scholar
  10. Borlongan, I.G. and Benitez, L.V. 1992. Lipid and fatty acid composition of milkfish (Chanos chanos Forsskal) grown in freshwater and seawater. Aquaculture 104: 79–89.Google Scholar
  11. Christie, W.W. 1982. Lipid Analysis, 2nd Edn. Pergamon Press, Oxford.Google Scholar
  12. Daikoku, T., Yano, I. and Masui, M. 1982. Lipid and fatty acid compositions and their changes in the different organs and tissues of guppy,Poecilia reticulata on sea water adaptation. Comp. Biochem. Physiol. 73A: 167–174.Google Scholar
  13. Dustan, J. and Knox, J.D.E. 1992. Acclimation of Atlantic salmon (Salmo salar) parr to seawater in autumn: stimulatory effect of a long photoperiod. Aquaculture 103: 341–358.Google Scholar
  14. Erdal, J.I., Evensen, Oe., Kaurstad, O.K., Lillehaug, A., Solbakken, R. and Thorud, K. 1991. Relationship between diet and immune response in Atlantic salmon (Salmo salar L.) after feeding various levels of ascorbic acid and omega-3 fatty acids. Aquaculture 98: 363–379.Google Scholar
  15. Finstad, B., Staurnes, M. and Reite, O.B. 1988. Effect of low temperature on sea-water tolerance in rainbow trout,Salmo gairdneri. Aquaculture 72: 319–328.Google Scholar
  16. Finstad, B., Nilssen, K.J. and Gulseth, O.A. 1989. Sea-water tolerance in freshwater-resident Arctic char (Salvelinus alpinus). Comp. Biochem. Physiol. 92A: 599–600.Google Scholar
  17. Folch, J., Lees, M. and Sloane Stanley, G.H. 1957. A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem. 226: 497–509.Google Scholar
  18. Hagve, T.A., Gronn, M. and Christophersen, B.O. 1991b. The decrease in osmotic fragility of erythrocytes durine supplementation with n-3 fatty acids is a transient phenomenon. Scand. J. Clin. Lab. Invest. 51: 493–495.Google Scholar
  19. Hagve, T.A., Johansen, Y. and Christophersen, B. 1991a. The effects of n-3 fatty acids on osmotic fragility of rat erythrocyte. Biochim. Biophys. Acta 1084: 251–254.Google Scholar
  20. Hansen, H.J.M. 1987. Comparative studies on lipid metabolism in various salt-transporting organs of the European eel (Anguilla anguilla). Monounsaturated phosphatidylethanolamine as a key substance. Comp. Biochem. Physiol. 88B: 323–332.Google Scholar
  21. Hansen, H.J.M., Olsen, A.G. and Rosenkilde, P. 1992. Comparative studies on lipid metabolism in salt-transporting organs of the rainbow trout (Oncorhynchus mykiss W) — further evidence of monounsaturated phosphatidylethanolamine as a key substance. Comp. Biochem. Physiol. 103B: 81–88.Google Scholar
  22. Hegab, S.A. and Hanke, W. 1982. Electrolyte changes and volume regulatory processes in the carp (Cyprinus carpio) during osmotic stress. Comp. Biochem. Physiol. 71A: 157–164.Google Scholar
  23. Henderson, R.J. and Tocher, D.R. 1987. The lipid composition and biochemistry of freshwater fish. Prog. Lipid Res. 26: 281–347.Google Scholar
  24. Hirano, T., Morisawa, M. and Suzuki, K. 1978. Changes in plasma and coelomic fluid composition of the mature salmon (Oncorhynchus keta) during freshwater adaptation. Comp. Biochem. Physiol. 61A: 5–8.Google Scholar
  25. Hirano, T., Ogasawara, T., Hasegawa, S., Iwata, M. and Nagahama, Y. 1990. Changes in plasma hormone levels during loss of hypoosmoregulatory capacity in mature chum salmon (Oncorhynchus keta) kept in seawater. Gen. Comp. Endocrinol. 78: 254–262.Google Scholar
  26. Hwang, P.P., Sun, C.M. and Wu, S.M. 1989. Changes of plasma osmolality, concentration and gill Na-K-ATPase activity in tilapiaOreochromis mossambicus during seawater acclimation. Mar. Biol. 100: 295–299.Google Scholar
  27. Lega, Y.V., Chernitsky, A.G. and Belkovsky, N.M. 1992. Effect of low sea water temperature on water balance in the Atlantic salmon, (Salmo salar L.). Fish Physiol. Biochem. 10: 145–148.Google Scholar
  28. Leray, C., Chapelle, S., Duportail, G. and Lorenz, A.F. 1984. Changes in the fluidity and 22:6(n-3) content in phospholipids of trout intestinal brush-border membrane as related to environmental salinity. Biochim. Biophys. Acta 778: 233–238.Google Scholar
  29. Morisawa, M., Hirano, T. and Suzuki, K. 1979. Changes in blood and seminal plasma composition of the mature salmon (Oncorhynchus keta) during adaptation to freshwater. Comp. Biochem. Physiol. 64B: 325–329.Google Scholar
  30. Nicholson, B.L. and Byrne, C. 1973. An established cell line from the Atlantic salmon (Salmo salar). J. Fish. Res. Bd. Can. 30: 913–916.Google Scholar
  31. Nonnotte, G. and Truchot, J.-P. 1990. Time course of extracellular acid-base adjustments under hypo- or hyperosmotic conditions in the euryhaline fishPlatichthys flesus. J. Fish Biol. 36: 181–190.Google Scholar
  32. Oguri, M. and Ooshima, Y. 1977. Early changes in the plasma osmolality and ionic concentrations of rainbow trout and goldfish following direct transfer from fresh-water to sea water. Bull. Jap. Soc. Sci. Fish. 43: 1253–1257.Google Scholar
  33. Rydevik, M., Borg, B., Haux, C., Kawauchi, H. and Bjornsson, B.Th. 1990. Plasma growth hormone levels increase during seawater exposure of sexually mature Atlantic salmon parr (Salmo salar L.). Gen. Comp. Endocrinol. 80: 9–15.Google Scholar
  34. Salte, R., Thomassen, M.S. and Wold, K. 1988. Do high levels of dietary polyunsaturated fatty acids (EPA/DHA) prevent diseases associated with membrane degeneration in farmed Atlantic salmon at low water temperatures. Bull. Eur. Ass. Fish Pathol. 8: 63–66.Google Scholar
  35. Schmidt-Nielsen, K. 1979. Animal Physiology: Adaptation and Environment. 2nd. Edition. Cambridge University Press, Cambridge.Google Scholar
  36. Sheridan, M.A. 1989. Alterations in lipid metabolism accompanying smoltification and seawater adaptation of salmonid fish. Aquaculture 82: 191–203.Google Scholar
  37. Sheridan, M.A., Allen, W.V. and Kerstetter, T.H. 1985. Changes in the fatty acid composition of steelhead trout,Salmo gairdnerii Richardson, associated with parr-smolt transformation. Comp. Biochem. Physiol. 80B: 671–676.Google Scholar
  38. Surette, M.E., Croset, M., Lokesh, B.R. and Kinsella, J.E. 1990. The fatty acid composition and Na+·K+ATPase activity of kidney microsomes from mice consuming diets of varying docosahexaenoic acid and linoleic acid ratios. Nutr. Res. 10: 211–218.Google Scholar
  39. Takeuchi, T., Kang, S.-J. and Watanabe, T. 1989. Effects of environmental salinity on lipid classes and fatty acid composition in gills of Atlantic salmon. Nippon Suisan Gakkaishi 55: 1395–1405.Google Scholar
  40. Tocher, D.R. and Dick, J.R. 1990a. Polyunsaturated fatty acid metabolism in cultured fish cells: Incorporation and metabolism of (n-3) and (n-6) series acids by Atlantic salmon (Salmo salar) cells. Fish Physiol. Biochem. 8: 311–319.Google Scholar
  41. Tocher, D.R. and Dick, J.R. 1990b. Incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids in phospholipid classes in cultured Atlantic salmon (Salmo solar) cells. Comp. Biochem. Physiol. 96B: 73–79.Google Scholar
  42. Tocher, D.R. and Harvie, D.G. 1988. Fatty acid compositions of the major phosphoglycerides from fish neural tissues; (n-3) and (n-6) polyunsaturated fatty acids in rainbow trout (Salmo gairdneri) and cod (Gadus morhua) brains and retinas. Fish Physiol. Biochem. 5: 229–239.Google Scholar
  43. Tocher, D.R. and Mackinlay, E.E. 1990. Incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids in phospholipid classes in cultured turbot (Scophthalmus maximus) cells. Fish Physiol. Biochem. 8: 251–260.Google Scholar
  44. Tocher, D.R. and Sargent, J.R. 1990. Effect of temperature on the incorporation into phospholipid classes and metabolismvia desaturation and elongation of n-3 and n-6 polyunsaturated fatty acids in fish cells in culture. Lipids 25: 435–442.Google Scholar
  45. Tocher, D.R., Carr, J. and Sargent, J.R. 1989. Polyunsaturated fatty acid metabolism in fish cells: Differential metabolism of (n-3) and (n-6) series acids by cultured cells originating from a freshwater teleost fish and from a marine teleost fish. Comp. Biochem. Physiol. 94B: 367–374.Google Scholar
  46. Tocher, D.R., Castell, J.D., Dick, J.R. and Sargent, J.R. 1994. Effects of salinity on the growth and lipid composition of Atlantic salmon (Salmo solar) and turbot (Scophthalmus maximus) cells in culture. Fish Physiol. Biochem. (In press).Google Scholar
  47. Tocher, D.R., Sargent, J.R. and Frerichs, G.N. 1988. The fatty acid compositions of established fish cell lines after long-term culture in mammalian sera. Fish Physiol. Biochem. 5: 219–227.Google Scholar
  48. Toneys, M.L. and Coble, D.W. 1980. Mortality, hematocrit, osmolality, electrolyte regulation, and fat depletion of young-of-the-year freshwater fishes under simulated winter conditions. Can. J. Fish. Aquat. Sci. 37: 225–232.Google Scholar
  49. Vitiello, F. and Zanetta, J.-P. 1978. Thin-layer chromatography of phospholipids. J. Chromat. 166: 637–640.Google Scholar
  50. Weirich, C.R. and Tomasso, J.R. 1991. Confinement- and transport-induced stress on red drum juveniles: effects of salinity. Progr. Fish Cult. 53: 146–149.Google Scholar
  51. Wertheimer, A.C. 1984. Maturation success of pink salmon (Oncorhynchus gorbuscha) and coho salmon (O. kisutch) held under three salinity regimes. Aquaculture 43: 195–212.Google Scholar
  52. Woo, N.Y.S. and Tong, W.C.M. 1982. Salinity adaptation in the snakehead,Ophiocephalus maculatus Lacepede: changes in oxygen consumption, branchial Na+-K+-ATPase and body composition. J. Fish Biol. 20: 11–19.Google Scholar
  53. Zuniga, M.E., Lokesh, B.R. and Kinsella, J.E. 1988. Effects of dietary N-6 and N-3 polyunsaturated fatty acids on composition and enzyme activities in liver plasma membrane of mice. Nutr. Res. 8: 1051–1060.Google Scholar

Copyright information

© Kugler Publications 1995

Authors and Affiliations

  • Douglas R. Tocher
    • 1
  • John D. Castell
    • 1
  • James R. Dick
    • 1
  • John R. Sargent
    • 1
  1. 1.NERC Unit of Aquatic Biochemistry, School of Natural SciencesUniversity of StirlingStirlingScotland

Personalised recommendations