|Appears in Collections:||eTheses from Faculty of Natural Sciences legacy departments|
|Title:||Influences of dietary polyunsaturated fatty acids on tissue fatty acid composition and eicosanoid production in Atlantic salmon (Salmo salar)|
|Authors:||Bell, John Gordon|
|Publisher:||University of Stirling|
|Abstract:||1. The literature has been reviewed with respect to the dietary intake and subsequent metabolism of polyunsaturated fatty acids (PUFA), of both the n-6 and n-3 series, in teleost fish. Particular emphasis has been made to the physiological roles of PUFA with respect to cell membrane function and eicosanoid production. 2. Atlantic salmon post-smolts were fed practical-type diets, based on fish meal, in three separate dietary experiments of 10-16 weeks duration. The first trial compared dietary lipid supplied either as fish oil (FO) or as sunflower oil (SO) with the diets having an n-3/n-6 PUFA ratio of 9.4 and 0.2 respectively. The second trial used diets formulated with blends of FO, SO, grape seed oil and safflower oil to provide linoleic acid at 10, 25 and 45% of total dietary fatty acids. The third trial was similar to the first but with an additional diet in which the lipid component was supplied by linseed oil (LO). All diets satisfied the nutritional requirements of salmonid fish for n-3 PUFA. There were no statistically significant differences in final weights between dietary treatments in the third trial. However, in the second trial fish fed the intermediate level of linoleic acid (25%) attained a significantly higher final weight compared to both other treatments while fish fed the highest level of linoleic acid (45%) had significantly lower final weights compared to both other treatments. In the first trial the effect of diet on growth (weight gain) could not be ascertained as the initial weights of the fish were significantly different. 3. A number of fish fed SO developed severe cardiac lesions which caused thinning of the ventricular wall and heart muscle necrosis. In addition the fish fed diets containing SO were susceptible to a transportation-induced shock syndrome that resulted in 30% mortality. 4. Incorporation of linoleic acid (18:2n-6) into membrane phospholipids increased in response to dietary intake with fish fed SO having increased levels of 18:2n-6 (up to 15-fold), 20:2n-6 (up to 12-fold), 20:3n-6 (up to 25-fold) and arachidonic acid (AA; 20:4n-6) (up to 3-fold), and decreased levels of eicosapentaenoic acid (EPA; 20:5n-3) (up to 3-fold). The ratio of n-3/n-6 PUFA was decreased (up to 4-fold) and the20:4n-6/20:5n-3 ratio increased (up to 9-fold) in membrane phospholipids from fish fed SO compared to those fed fish oil. While the tissue phospholipids from fish fed La had increased levels of 18:2n-6, 20:2n-6 and 20:3n-6, the levels of AA, 22:4n-6 and 22:5n-6 were similar to or significantly reduced compared to fish fed FO. Membrane phospholipids from fish fed LO also had increased 18:3n-3 and 20:4n-3 compared to both other treatments while in some tissues and phospholipid classes EPA was increased compared to fish fed FO. 5. These dietary induced changes in phospholipid eicosanoid precursor ratio were reflected in altered eicosanoid production. In gill cells, stimulated with the calcium ionophore A23187, 12-hydroxy-8, 10, 14, 17-eicosapentaenoic acid (12-HEPE) was the major 12-lipoxygenase product in fish fed Fa. In stimulated gill cells from fish fed SO and LO, 12-HEPE, 12-hydroxy-5, 8, 10, 14-eicosatetraenoic acid (12-HETE), 14- hydroxy-4, 7, 10, 13, 16, 19-docosahexaenoic acid (14-HDHE) and thromboxane B2 (TXB2) were all decreased compared to fish fed FO. However, the ratio of 12- HETE/12-HEPE was significantly elevated in stimulated gill cells from SO-fed fish compared to both other treatments. In stimulated blood leucocytes leukotriene B4 (LTB4)' 12-HETE and TXB2 were significantly increased while LTB5 and 12-HEPE were significantly decreased in fish fed SO compared to those fed FO. Blood leucocytes from fish fed LO produced less TXB2 compared to fish fed SO and prostaglandin E2 was reduced compared to both other treatments. In isolated cardiac myocytes stimulated with A23187, TXB2 production was increased in SO fed fish compared to those fed FO. 6. The activity of cardiac sarcoplasmic reticulum Ca2+-Mg2+ATPase was not affected by dietary treatment. 7. An established cell line derived from chum salmon heart (CHH-1) was utilised to study PUFA metabolism. The CHH-1 cells exhibited considerable A6 desaturase activity but showed no preference towards n-3 over n-6 PUFA. CHH-1 cells did exhibit significant A5 desaturase activity which showed a preference towards n-3 PUFA. No A4 desaturation activity was observed. Elongation of C20 PUFA was especially active in CHH-1 cells with C22 PUFA being specifically incorporated into phosphatidylethanolamine (PE) and phosphatidylserine (PS). CHH-1 cells supplemented with 20:3n-6 showed reduced growth rate, cell death and unusual pycnotic appearance, compared to those supplemented with other PUFA. 8. The lipid compositions of hearts and livers from wild and farmed parr and presmolts were analysed and compared. The fatty acid compositions of triacylglycerols (TAG) and phospholipids from both farmed parr and pre-smolts contained greater amounts of monoenoic fatty acids compared to their wild counterparts. TAG, phosphatidylcholine (PC) and PE from heart and liver of wild fish contained more 18:2n-6 and AA compared to farmed fish. Linolenic acid, EPA and 22:Sn-3 were increased in hearts and livers of wild fish compared to farmed. Docosahexaenoic acid (DHA; 22:6n-3) levels were higher in heart and liver of farmed fish, particularly in heart PC, PS and TAG. The n-3/n-6 PUFA ratio was generally lower in wild compared to farmed fish, largely due to higher n-6 PUFA, in particular AA, in wild fish. 9. The results are discussed with respect to the competitive interactions between PUFA of the n-6 and n-3 series which determine the fatty acid compositions of membrane phospholipids in salmon. The ratio of n-3/n-6 PUFA in membrane phospholipids, and in particular the ratio of AAIEPA, appears important in terms of membrane physiology and biochemistry, eicosanoid production and the development of cardiac histopathological lesions.|
|Type:||Thesis or Dissertation|
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