Duplicate groups of Atlantic salmon post-smolts were fed five practical-type diets in which the added lipid was 100% fish oil [FO; 0% rapeseed oil (0% RO)], 90% FO + 10% RO (10% RO), 75% FO + 25% RO (25% RO), 50% FO + 50% RO (50% RO) or 100% RO, for a period of 17 wk. There were no effects of diet on growth rate or feed conversion nor were any histopathological lesions found in liver, heart, muscle or kidney. The greatest accumulation of muscle lipid was in fish fed 0% RO, which corresponded to significantly lower muscle protein in this group. The highest lipid levels in liver were found in fish fed 100% RO. Fatty acid compositions of muscle lipid correlated with RO inclusion in that the proportions of 18:1(n-9), 18:2(n-6) and 18:3(n-3) all increased with increasing dietary RO (r = 0.98-1.00, P < 0.013). The concentrations of eicosapentaenoic acid [20:5(n-3)] and docosahexaenoic acid [22:6(n-3)] in muscle lipid were significantly reduced (P < 0.05), along with total saturated fatty acids, with increasing dietary RO. Diet-induced changes in liver fatty acid compositions were broadly similar to those in muscle. Hepatic fatty acid desaturation and elongation activities, measured using [1-(14)C] 18:3(n-3), were increased with increasing dietary RO. Limited supplies of marine fish oils require that substitutes be found if growth in aquaculture is to be maintained such that fish health and product quality are not compromised. Thus, RO can be used successfully as a substitute for fish oil in the culture of Atlantic salmon in sea water although at levels of RO >50% of dietary lipid, substantial reductions occur in muscle 20:5(n-3), 22:6(n-3) and the (n-3)/(n-6) polyunsaturated fatty acid (PUFA) ratio, which will result in reduced availability of the (n-3) highly unsaturated fatty acids that are beneficial for human health.

Supplies of marine fish oils (FO) are limited and continued growth in aquaculture production dictates that substitutes must be found that do not compromise fish health and product quality. In this study the suitability of crude palm oil (PO) as a replacement for FO in diets of Atlantic salmon was investigated. Duplicate groups of Atlantic salmon post-smolts were fed four practical-type diets in which the added lipid was either 100% FO and 0% crude PO (0% PO); 75% FO and 25% PO (25% PO); 50% FO and 50% PO (50% PO); and 100% PO, for 30 wk. There were no effects of diet on growth rate or feed conversion ratio nor were any histopathological lesions found in liver, heart or muscle. Lipid deposition was greatest in fish fed 0% PO and was significantly greater than in fish fed 50% and 100% PO. Fatty acid compositions of muscle total lipid were correlated with dietary PO inclusion such that the concentrations of 16:0, 18:1(n-9), 18:2(n-6), total saturated fatty acids and total monoenoic fatty acids increased linearly with increasing dietary PO. The concentration of eicosapentaenoic acid [20:5(n-3)] was reduced significantly with increasing levels of dietary PO but the concentration of docosahexaenoic acid [22:6(n-3)] was significantly reduced only in fish fed 100% PO, compared with the other three treatments. Similar diet-induced changes were seen in liver total lipid fatty acid compositions. Hepatic fatty acid desaturation and elongation activities were approximately 10-fold greater in fish fed 100% PO than in those fed 0% PO. This study suggests that PO can be used successfully as a substitute for FO in the culture of Atlantic salmon in sea water. However, at levels of PO inclusion above 50% of dietary lipid, significant reductions in muscle 20:5(n-3), 22:6(n-3) and the (n-3):(n-6) PUFA ratio occur, resulting in reduced availability of these essential (n-3) highly unsaturated fatty acids to the consumer.

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Abbreviations: AA, arachidonic acid; BHT, butylated hydroxytoluene; DHA, docosahexaenoic acid; EFA, essential fatty acid; EPA, eicosapentaenoic acid; FAF-BSA, fatty acid free bovine serum albumin; FO, fish oil; HBSS, Hanks balanced salt solution; HUFA, highly unsaturated fatty acids acids (carbon chain length ≥ C 20 with ≥ 3 double bonds); PL, polar lipid; PUFA, polyunsaturated fatty acids; VO, vegetable oil; TLC, thin-layer chromatography. 2 AbstractThe desaturation and elongation of [1-14 C]18:3n-3 was investigated in hepatocytes from different populations and three different species of salmonids indigenous to Scotland, brown trout, Atlantic salmon and Arctic charr. Two groups of fish were sampled, before and after they were fed two experimental diets, a control diet containing fish oil and a diet containing vegetable oil (a 1:1 blend of linseed oil and rapeseed oil) for 12 weeks. At each sampling time, fatty acyl desaturation and elongation activity was determined in isolated hepatocytes and samples of liver were also collected for lipid compositional analysis. At the initiation of the dietary trial, the liver polar lipid fatty acid compositions of salmon and brown trout were very similar to each other, and the two charr populations were similar to each other having lower total n-3 polyunsaturated fatty acids (PUFA) and 22:6n-3, but higher 20:5n-3 than the other salmonids. Initially, hepatocyte desaturation activity varied with the highest activity in brown trout followed by salmon then charr. Production of 20:5n-3 was particularly high in brown trout. Desaturation of [1-14 C]18:3n-3 was significantly greater in all fish fed the diet containing vegetable oil compared to fish fed the diet containing fish oil. The increase in activity was less in brown trout compared to the other groups of fish. Feeding the vegetable oil diet increased the levels of 18:2n-6, 20:3n-6, total n-6PUFA, 18:3n-3, 18:4n-3, 20:3n-3 and 20:4n-3, and decreased 22:6n-3 and the n-3/n-6 ratio in salmon and brown trout. By contrast, in charr fed the vegetable oil diet, there was no increase in 18:3n-3, 18:4n-3, 20:3n-3 or 20:4n-3 in liver polar lipid and the level of 22:6n-3 was not decreased. In addition, there was only a modest increase in the levels of 18:2n-6 and total n-6PUFA and so the n-3/n-6 ratio was only slightly decreased. The percentage of 20:4n-6, which was not increased in salmon and brown trout fed vegetable oil, was increased in charr fed the vegetable oil diet. Overall, the results indicated that there were significant differences in liver PUFA metabolism between Arctic charr and the other salmonids that could have important consequences, both physiologically and in their ability to be successfully cultured on diets containing vegetable oils.Running title: PUFA composition and desaturation in salmonids.

Samples of steaks from market-size salmon (2.5-5 kg), produced under the requirements of the Product Certification Scheme for Scottish Quality Farmed Salmon, were obtained from five Scottish salmon producers on a weekly basis for over 2 years. Samples were assayed for total lipid content, lipid class composition, fatty acid composition, vitamin E content, and carotenoid pigment [astaxanthin (AX) and canthaxanthin (CX)] content. In addition, samples were obtained from the same producers to assess loss of carotenoid pigment in the period up to 120 h post-harvest and to measure lipid and pigment content of different body regions. The analyses showed a positive correlation between dietary lipid and deposition of lipid in flesh, although there was great variation in flesh lipid content within each dietary lipid level. The average lipid content of Scottish salmon was 10.1 +/- 2.9% (n = 495). Salmon flesh was rich in the n-3 highly unsaturated fatty acids, 22:6n-3 and 20:5n-3, with average values of 11.3 and 5.4% of total fatty acids or 10.2 and 4.8 g/kg flesh, respectively. Vitamin E content of salmon flesh was around 30 mg/kg but was significantly higher in fish fed the lowest dietary lipid level. Studies investigating changes in carotenoid pigmentation up to 120 h post-harvest suggested that fish fed AX alone showed a loss of pigment over time that did not occur in fish fed either CX alone or a combination of AX and CX. Measurement of lipid content of different body regions showed that the highest levels were found in the region immediately in front of the dorsal fin and that the lowest lipid levels were in the tail region. Total lipid content of all five body zones was positively correlated with dietary lipid. Carotenoid pigment levels were the same across all body zones and were not affected by dietary lipid.

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Changes in fatty acid metabolism in Atlantic salmon (Salmo salar) induced by vegetable oil (VO) replacement of fish oil (FO) and high dietary oil in aquaculture diets can have negative impacts on the nutritional quality of the product for the human consumer, including altered flesh fatty acid composition and lipid content. A dietary trial was designed to investigate the twin problems of FO replacement and high energy diets in salmon throughout the entire production cycle. Salmon were grown from first feeding to around 2 kg on diets in which FO was completely replaced by a 1:1 blend of linseed and rapeseed oils at low (14-17%) and high (25-35%) dietary oil levels. This paper reports specifically on the influence of diet on various aspects of fatty acid metabolism. Fatty acid compositions of liver, intestinal tissue and gill were altered by the diets with increased proportions of C 18 polyunsaturated fatty acids and decreased proportions of n-3 highly unsaturated fatty acids (HUFA) in fish fed VO compared to fish fed FO. HUFA synthesis in hepatocytes and enterocytes was significantly higher in fish fed VO, whereas β-oxidation was unaltered by either dietary oil content or type. Over the entire production cycle, HUFA synthesis in hepatocytes showed a decreasing trend with age interrupted by a large peak in activity at seawater transfer. Gill cell prostaglandin (PG) production showed a possible seasonal trend, with peak activities in winter and low activities in summer and at seawater transfer. PG production in seawater was lower in fish fed the high oil diets with the lowest PG production generally observed in fish fed high VO. The changes in fatty acid metabolism induced by high dietary oil and VO replacement contribute to altered flesh lipid content and fatty acid compositions, and so merit continued investigation to minimize any negative impacts that sustainable, environmentally-friendly and cost-effective aquaculture diets could have in the future.Abbreviations: FO, fish oil; HUFA, highly unsaturated fatty acids acids (carbon chain length ≥ C 20 with ≥ 3 double bonds); LO, linseed oil; RO, rapeseed oil; VO, vegetable oil.3

Duplicate groups of Atlantic salmon parr were fed diets containing either fish oil (FO), rapeseed oil (RO), linseed oil (LO) or linseed oil supplemented with arachidonic acid (20:4n-6; AA) (LOA) from October (week 0) to seawater transfer in March (week 19). From March to July (weeks 20-34) all fish were fed a fish oil-containing diet. Fatty acyl desaturation and elongation activity in isolated hepatocytes incubated with [1-14 C]18:3n-3 increased in all dietary groups, peaking in early March about one month prior to seawater transfer. Desaturation activities at their peak were significantly greater in fish fed the vegetable oils, particularly RO, compared to fish fed FO. Docosahexaenoic acid (22:6n-3:DHA) and AA in liver and gill polar lipids (PL) increased in all dietary groups during the freshwater phase whereas eicosapentaenoic acid (20:5n-3;EPA) increased greatly in all groups after seawater transfer. The AA/EPA ratio in tissue PL increased up to seawater transfer and then decreased after transfer. AA levels and the AA/EPA ratio in gill PL were generally higher in the LOA group. The levels of 18:3n-3 in muscle total lipid were increased significantly in the LO, LOA and, to a lesser extent, RO groups prior to transfer but were reduced to initial levels by the termination of the experiment (week 34). In contrast, 18:2n-6 in muscle total lipid was significantly increased after 18 weeks in fish fed the diets supplemented with RO and LO, and was significantly greater in the FO and RO groups at the termination of the experiment. Gill PGF production showed a large peak about two months after transfer to seawater. The production of total PGF post-transfer was significantly lower in fish previously fed the LOA diet. However, plasma chloride concentrations in fish subjected to a seawater challenge at 18 weeks were all lower in fish fed the diets with vegetable oils. This effect was significant in the case of fish receiving the diet with LOA, compared to those fed the diet containing FO. The present study showed that during parr-smolt transformation in Atlantic salmon there is a pre-adaptive increase in hepatocyte fatty acyl desaturation/elongation activities that is controlled primarily by environmental factors such as photoperiod and temperature but that can also be significantly modulated by diet. Feeding salmon parr diets supplemented with rapeseed or linseed oils prevented inhibition of the desaturase activities that is induced by feeding parr diets with fish oils and thus influenced the smoltification process by altering tissue PL fatty acid compositions and eicosanoid production. These effect, in turn, had a beneficial effect on the ability of the fish to osmoregulate and thus adapt to salinity changes.