Replacing dietary fish oil with increasing levels of rapeseed oil and olive oil – effects on Atlantic salmon (Salmo salar L.) tissue and lipoprotein lipid composition and lipogenic enzyme activities

Six groups of Atlantic salmon, initial weight 142 ± 1 g, were fed increasing dietary inclusion of rapeseed oil (RO) in a regression design and one group was fed a 50% olive oil/50% capelin oil diet. Fatty acid composition was measured in red and white muscle, liver, and fatty acid and lipid class composition was measured in plasma and in the lipoproteins; very low density lipoprotein, low density lipoprotein, high density lipoprotein and nonlipoprotein fraction after 22 and 42 weeks of feeding. Further, the activities of liver NADH‐isocitrate dehydrogenase (ICDH), malic enzyme, glucose‐6‐phosphate dehydrogenase (G6PDH) and 6‐phosphogluconate dehydrogenase were measured at each sampling point. After 42 weeks of feeding the experimental diets, the tissue and lipoprotein fatty acid composition was highly affected by dietary fatty acid composition. Regressions showed that 22:1n ? 11, 18:1n ? 9, 18:3n ? 3 and 18:2n ? 6 are readily metabolized in all tissues analysed. Further, 20:5n ? 3 seems to be metabolized in muscle and retained in liver. 22:6n ? 3 was selectively retained in all the analysed tissues, and with higher retention in liver and plasma with higher polar lipid/neutral lipid ratio compared to white and red muscle. Liver from salmon fed 100% RO showed decreased G6PDH and increased ICDH activities compared to the other dietary groups; however, no linear relationship related to increased RO inclusion was detected. The amount of plasma lipoproteins, liver monoene fatty acid level and lipogenic enzyme activity decreased from the autumn to the winter sampling with concomitant decrease in temperature.     

Gene expression of fatty acid-binding proteins, fatty acid transport proteins (cd36 and FATP) and β-oxidation-related genes in Atlantic salmon (Salmo salar L.) fed fish oil or vegetable oil

Relative gene expression pattern of fatty acid transport proteins (FATP and cd36), intracellular fatty acid-binding proteins (FABP3, FABP10 and FABP11), β-oxidation-related genes [carnitine palmitoyl transferase II (CPTII), peroxisome proliferator-activated receptor β (PPARβ), acyl-CoA oxidase (AOX), long-chain fatty acyl-CoA synthetase (FACS), acyl-CoA dehydrogenase (dehydrogenase)] and uncoupling protein 2 (UCP2) was assessed by RT-qPCR in Atlantic salmon muscle (red and white), liver, heart, myosepta and visceral fat. FABP11, a FABP isoform not previously described in Atlantic salmon, was highly expressed in visceral fat and myosepta and at the lower level in red muscle, white muscle, myosepta and heart. Furthermore, Atlantic salmon were fed either a diet containing fish oil (FO) or a complete replacement of FO with a vegetable oil blend (55% rapeseed oil, 30% palm oil and 15% linseed oil; VO) for the production cycle (27 months from start of feeding and until ∼4.5 kg mean weight). The expression of genes related to β-oxidation, fatty acid uptake and transport in the white muscle indicate ( n  = 3) significant down-regulation in VO fed Atlantic salmon and correlated with previously reported white muscle triacylglycerol stores and β-oxidation. FABP11 in visceral fat and myosepta was also down-regulated in VO fed fish.     

Replacement of a large portion of fish oil by vegetable oils does not affect lipogenesis, lipid transport and tissue lipid uptake in European seabass (Dicentrarchus labrax L.)

In order to investigate the impact of dietary lipid sources on mechanisms involved in lipid deposition, three groups of European seabass fingerlings with average initial body weight of 5.2 ± 1.0 g were fed three diets differing only by lipid source. These diets were: 100% anchovy oil (diet A), 40% anchovy oil-60% mix of vegetable oils (35% linseed, 15% palm, 10% rapeseed) (diet B) and 40% anchovy oil-60% mix of vegetable oils (24% linseed, 12% palm, 24% rapeseed) (diet C). After 64 weeks of rearing, when seabass reached the size of 160 g, the activity of lipogenic enzymes (fatty acid synthetase, glucose-6-phosphate dehydrogenase and malic enzyme) in liver and of lipoprotein lipase (LPL) in perivisceral adipose tissue, liver and white muscle were measured. Transport of lipid by lipoproteins was examined by determining plasma lipid composition and lipoprotein classes. Dietary oil source did not modify growth performance or lipid content of flesh and liver of seabass. Replacement of 60% of fish oil by the two mixtures of vegetable oils had no significant effect on hepatic lipogenesis and activity of LPL in liver and adipose tissue. Activity of LPL in white muscle was decreased in fish fed diet C compared to those fed diets A and B. Diets containing the mixture of vegetable oils led to lowered plasma, VLDL and LDL cholesterol levels compared to diet A.It is concluded that replacing 60% of fish oil by the two mixtures of vegetable oils in the feeds of European seabass fingerlings until they reach the size of 160 g has no marked effect on growth performance, lipogenesis and tissue lipid uptake but has a hypocholesterolemic effect.     

Effects of dietary lipid level and vegetable oil on fatty acid metabolism in Atlantic salmon (Salmo salar L.) over the whole production cycle

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 C₁₈ 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. This revised version was published online in July 2006 with corrections to the Cover Date.     

Efficacy of an equal blend of canola oil and poultry fat as an alternate dietary lipid source for Atlantic salmon (Salmo salar L.) in seawater. II: effects on haematology and immunocompetence

This research examined the haematological and immunological responses of quadruplicate groups of juvenile (～400 g initial weight) Atlantic salmon (Salmo salar L.) that had each been fed daily to satiation for 12 weeks one of three high‐energy extruded diets of identical composition except for the supplemental dietary lipid (234.7 g kg^?1) source. The three experimental diets varied in the composition of supplement lipid; diet 1 contained 100% anchovy oil (AO), while diets 2 and 3 replaced 29.8% and 59.7% of the AO (respectively) with a 1:1 blend of canola oil (CO) and poultry fat (PF). Immediately following the feeding trial, a random sample of fish from each diet was sampled for determination of baseline levels of various haematological and immunological parameters. Thereafter, duplicate diet groups were vaccinated (against Listonella anguillarum) and reared on their respective experimental diets for an additional 4 weeks. At that time, the remaining fish were sampled similarly, and the different parameters were measured again. Comparisons between the different diet treatment groups were made before and after vaccination. There were no significant diet treatment effects at either sample time, for haematocrit, differential leucocyte counts, erythrocyte counts, serum hemolytic activity or head kidney leucocyte respiratory burst activity. The fish fed diet 1 however, did show significantly higher post‐vaccination levels of peripheral blood leucocyte respiratory burst activity and higher serum antibody titres against L. anguillarum. The results suggest that the relatively low n‐6/n‐3 fatty acid ratios in the muscle and presumably other tissues of fish fed diet 1, may have resulted in a reduced production of immunocompromising eicosanoids than were produced in fish ingesting the other two diets that were based in part on the different amounts of the CO and PF blend. Long‐term studies are required to confirm this possibility.     

Efficacy of an equal blend of canola oil and poultry fat as an alternate dietary lipid source for Atlantic salmon (Salmo salar L.) in sea water. I: effects on growth performance, and whole body and fillet proximate and lipid composition

This study assessed the suitability and cost efficacy of an equal blend of canola oil (CO) and poultry fat (PF) as a supplemental dietary lipid source for juvenile Atlantic salmon. Quadruplicate groups of Atlantic salmon (～400 g) held in 4000 L outdoor fibreglass tanks supplied with running (35–40 L min^?1), aerated (dissolved oxygen, 7.88–10.4 mg L^?1), ambient temperature (8.6–10.9°C) sea water (salinity, 26–35 g L^?1) were fed twice daily to satiation one of three extruded dry pelleted diets of equivalent protein (488–493 g kg^?1 dry matter) and lipid (267–274 g kg^?1 dry matter) content for 84 days. The diets were identical in composition except for the supplemental lipid (234.7 g kg^?1) source viz., 100% anchovy oil (AO; diet COPF‐0), 70.2% AO and 29.8% CO and PF (diet COPF‐30), and 40.3% AO and 59.7% CO and PF (diet COPF‐60). Atlantic salmon growth rate, feed intake, feed efficiency, protein and gross energy utilization, percent survival and whole body and fillet proximate compositions were not affected by diet treatment. Cost per kilogram weight gain was about 10% less for fish fed diet COPF‐60 than for diet COPF‐0. Percentages of saturated fatty acids in dietary and fillet lipids varied narrowly. Moreover, percentages of 18:1n‐9, monounsaturated fatty acids, 18:2n‐6, n‐6 fatty acids, 18:3n‐3, and ratios of n‐6 to n‐3 fatty acids in the flesh lipids were directly related to the dietary level of CO and PF whereas 22:6n‐3, the total of 20:5n‐3 (eicosapentaenoic acid; EPA) and 22:6n‐3 (docosahexaenoic acid; DHA), and n‐3 fatty acids revealed the opposite trend. Percentages of 22:6n‐3, EPA and DHA, and n‐3 fatty acids were significantly depressed in fish fed diet COPF‐60 versus diet COPF‐0. We conclude that a 1:1 blend of CO and PF is an excellent cost‐effective dietary source of supplemental lipid for Atlantic salmon in sea water.     

Effects of substitution of dietary fish oil with a blend of vegetable oils on liver and peripheral blood leucocyte fatty acid composition, plasma prostaglandin E2 and immune parameters in three strains of Atlantic salmon (Salmo salar)

Duplicate groups of three genetic strains of Atlantic salmon smolts were cultured on diets containing either fish oil (FO) or a blend of vegetable oils (VO). Fatty acid compositions of liver and peripheral blood leucocytes were compared. The effect of different strains and diets on innate immune parameters and plasma prostaglandin E₂ were also measured. Two strains were selected as being either 'fat' or 'lean' in terms of muscle adiposity. The third strain was a commercial stock (MH). Total replacement of dietary FO with VO resulted in reduced docosahexaenoic (DHA; 22:6 n -3) and eicosapentaenoic acids (EPA; 20:5 n -3) in liver, while oleic (18:1 n -9), linoleic (18:2 n -6) and α-linolenic (18:3 n -3) acids were all increased in VO-fed fish. Fatty acid compositions of blood leucocytes showed similar changes. Evaluation of innate immune function showed that in the fat strain, circulating leucocytes were significantly lower in VO fish. The lean strain also had significantly higher serum lysozyme activity than MH fish. Reduced haematocrit was seen in VO lean fish compared with FO lean fish. This study provides evidence of strain-induced differences in liver and leucocyte fatty acid compositions and innate immunity in Atlantic salmon fed either FO- or VO-based diets.     

Effects of dietary protein, and fat level and rapeseed oil on growth and tissue fatty acid composition and metabolism in Atlantic salmon (Salmo salar L.) reared at low water temperatures

A 12‐week feeding trial was conducted to elucidate the interactive effects of dietary fat, protein contents and oil source on growth, whole body proximate composition, protein productive value (PPV) and fatty acid (FA) composition of muscle and liver in Atlantic salmon (Salmo salar L.)` at low water temperatures (4.2 °C). Triplicate groups of Atlantic salmon (initial weight 1168 g) were fed six isoenergetic diets, formulated to provide either 390 g kg⁻¹ protein and 320 g kg⁻¹ fat (high‐protein diets) or 340 g kg⁻¹ protein and 360 g kg⁻¹ fat (low‐protein diets). Within each dietary protein/fat level, crude rapeseed oil (RO) comprised 0, 30 or 60% (R0, R30, R60, respectively) of the added oil. After 12 weeks, the overall growth and feed conversion ratio (FCR) were very good for all treatments [thermal growth coefficient (TGC): 4.76 (±0.23); FCR: 0.85 (±0.02)]. Significant effects were shown owing to the oil source on specific growth rate and TGC only. The liver and muscle FA compositions were highly affected by the graded inclusion of RO. The PPV was significantly affected by the dietary protein level. The results of this study suggest that more sustainable, lower protein diets with moderate RO inclusion can be used in Atlantic salmon culture at low water temperatures with no negative effects on growth and feed conversion, no major detrimental effects on lipid and FA metabolism and a positive effect on protein sparing.     

Temperature modulates liver lipid accumulation in Atlantic salmon (Salmo salar L.) fed low dietary levels of long‐chain n‐3 fatty acids

Atlantic salmon (Salmo salar) were fed five graded levels of eicosapentaenoic acid (EPA, 20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3), from 1.4 to 5.2% of total fatty acids (FA, 5–17 mg kg^?1 feed), and grew from ~160 g to ~3000 g, with the period from 1450 g onwards conducted both at 6 °C and at 12 °C. All fish appeared healthy, and there were no diet‐related differences in haematological or plasma parameters, as well as intestinal histological or gut microbiota analysis. Fish reared at 6 °C had higher accumulation of storage lipids in the liver compared to fish reared at 12 °C. Liver lipids also increased with decreasing dietary EPA + DHA at 6 °C, while there was no such relationship at 12 °C. Gene expression of SREBP1 and 2, LXR, FAS and CPT1 could not explain the differences in liver lipid accumulation. In liver polar lipids, DHA was found to be reduced when dietary EPA + DHA was <2.7% of FAs, while the level of EPA in the membranes was not affected. In conclusion, reducing dietary EPA + DHA from 5.2 to 1.4% of total FAs had a minor impact on fish health. Temperature was the factor that most affected the liver lipid accumulation, but there was also an interaction with dietary components.     

Partial substitution of fish oil with rapeseed, linseed and olive oils in diets for European sea bass (Dicentrarchus labrax L.): effects on flesh fatty acid composition, plasma prostaglandins E2 and F2α, immune function and effectiveness of a fish oil finishing diet

Triplicate groups of European sea bass (Dicentrarchus labrax L.), of initial weight 90 g, were fed four practical‐type diets in which the added oil was 1000 g kg^?1 fish oil (FO) (control diet), 600 g kg^?1 rapeseed oil (RO) and 400 g kg^?1 FO, 600 g kg^?1 linseed oil (LO) and 400 g kg^?1 FO, and 600 g kg^?1 olive oil (OO) and 400 g kg^?1 FO for 34 weeks. After sampling, the remaining fish were switched to the 1000 g kg^?1 FO diet for a further 14 weeks. Fatty acid composition of flesh total lipid was influenced by dietary fatty acid input but specific fatty acids were selectively retained or utilized. There was selective deposition and retention of docosahexaenoic acid (DHA; 22:6n‐3). Eicosapentaenoic acid (EPA; 20:5n‐3) and DHA were significantly reduced and linolenic (LNA; 18:3n‐3), linoleic (LA; 18:2n‐6) and oleic (OA; 18:1n‐9) acids significantly increased in flesh lipids following the inclusion of 600 g kg^?1 RO, LO and OO in the diets. No significant differences were found among different treatments on plasma concentrations of prostaglandin E₂ and prostaglandin F_2α. Evaluation of non‐specific immune function, showed that the number of circulating leucocytes was significantly affected (P < 0.001), as was macrophage respiratory burst activity (P < 0.006) in fish fed vegetable oil diets. Accumulation of large amounts of lipid droplets were observed within the hepatocytes in relation to decreased levels of dietary n‐3 HUFA, although no signs of cellular necrosis was evident. After feeding a FO finishing diet for 14 weeks, DHA and total n‐3 HUFA levels were restored to values in control fish although EPA remained 18% higher in control than in the other treatments. This study suggests that vegetable oils such as RO, LO and OO can potentially be used as partial substitutes for dietary FO in European sea bass culture, during the grow out phase, without compromising growth rates but may alter some immune parameters.     

Immune response and disease resistance in Atlantic salmon (Salmo salar L.) fed three levels of dietary vitamin E and the effect of vaccination on the liver status of antioxidant vitamins

Atlantic salmon were fed 40, 300 or 1100 mgall-rac--tocopheryl acetate kg feed^–1 for 12 weeks.After 6 weeks half of the fish in each group were marked and vaccinated (i.p.injection) against furunculosis and vibriosis. Liver -tocopherol levelsreflected the dietary input after 6 and 12 weeks of feeding. Noimmunomodulatoryeffects of dietary vitamin E on baseline levels of the immune parameters beforevaccination were detected in this study, as evaluated by antibody dependent andspontaneous plasma complement activities. In general, vaccination increasedplasma complement activities and the number of antigen specific antibodyproducing cells as compared with unvaccinated control fish, but with nodifferences with respect to the vitamin E regimes. Also the ability ofunvaccinated fish to withstand experimental furunculosis was unaffected bydietary vitamin E. The concentrations of -tocopherol and ascorbic acidinthe liver were, however, negatively affected 6 weeks post vaccination comparedto respective unvaccinated fish.     

High dietary 18:2n‐6/18:3n‐3 ratio does not inhibit elongation and desaturation of 18:3n‐3 to EPA and DHA in Atlantic salmon (Salmo salar L.)

This study investigated whether retention of n‐3 fatty acids (FAs) is influenced by the levels of dietary monounsaturated (MUFA)‐, saturated (SFA)‐ and n‐6 FAs. The feeding trial used a mixture design, with rapeseed‐, palm (PO)‐ and soybean oil (SO), providing high levels of MUFA, SFA and n‐6 FAs, respectively, while 18:3n‐3, eicosapentaenoic acid (20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3) were kept constant. Furthermore, a diet group with high SO and reduced 18:3n‐3 was included in addition to the mixture design. There were no differences in growth or proximate composition, but the PO diet gave reduced FA digestibility and increased feed conversion ratio. All diet groups had a net production of DHA, shown by retention values >100% (133%–177%). High dietary n‐6 FAs of up to 43% of FAs (18:2n‐6/18:3n‐3 ratio 8.6) did not negatively affect DHA retention, but rather had a small positive impact. High levels of substrate (dietary 18:3n‐3 at ~10% of FAs) did not enhance DHA retention compared to 5% 18:3n‐3. This is a highly significant finding for the aquaculture industry, allowing for greater flexibility in the choice of lipid sources to replace fish oil in salmon feeds.     

Effects of dietary levels of 20:5n-3 and 22:6n-3 on tissue lipid composition in juvenile Atlantic salmon, Salmo salar, with emphasis on brain and eye

Four dietary groups of juvenile Atlantic salmon, Salmo salar L., each with three replicates, were fed diets with increasing levels of docosahexaenoic acid (22:6n-3; DHA) and eicosapentaenoic acid (20:5n-3; EPA). Fatty acid composition of brain and eye was determined at the start and approximately every 3 weeks during the experimental period, and fatty acid composition of liver and fillet was determined in fish from the final sampling. Lipid class composition of brain and eye, and fatty acid composition of these lipid classes was determined at the end of the experiment. There was no effect of increasing dietary DHA content on fatty acid composition, lipid class composition or DHA levels in the lipid classes in the juvenile Atlantic salmon brain. The increasing dietary EPA content, however, was reflected in both the total fatty acid composition and in the EPA content in neutral lipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). A minor effect of the increasing dietary DHA content was found in the lipid composition of the juvenile salmon eye. Both EPA and 18:2n-6 levels in eye, however, clearly reflected the increasing and decreasing, respectively, dietary levels of these two fatty acids. The dietary EPA levels also affected the EPA levels in neutral lipids, PC, PE, PI and PS (phosphatidylserine) in the juvenile salmon eye. The results demonstrate that these dietary levels of DHA had no effect on brain lipid composition and only a minor effect on eye lipid composition. Furthermore, the dietary EPA levels significantly affected the lipid composition of both brain and eye. The fillet fatty acid composition reflected the dietary fatty acid composition, except for the DHA/EPA ratio, which was reversed in fillet compared with that in the diets. The liver fatty acid composition was also affected by the increasing dietary EPA and DHA levels.     

Effects of total fish oil replacement to vegetable oils at two dietary lipid levels on the growth,body composition,haemato‐immunological and serum biochemical parameters in caspian brown trout (Salmo trutta caspius Kessler, 1877)

This research aimed to evaluate the effects of two dietary fat levels [low fat (LF) (10%), high fat (HF) (20%)] and sources [fish oil (FO), vegetable oil (VO)] on the growth and some physiological parameters of Caspian brown trout fingerlings for 60 days. Tuna oil or blends of canola and soybean oils (85:15) were added to diets to design four feeds namely LFFO, HFFO, LFVO and HFVO according to the fat levels and sources. The fish fed the LFFO diet had lower weight gain than the other fish (P<0.05). The total n‐6 fatty acids increased in fish fed diets with the blends of VO, while the total n‐3 fatty acids decreased in these fish (P<0.05). Serum lysozyme activity was higher in fish fed the HFVO diet than the other fish (P<0.05). Serum glucose, total cholesterol, triglyceride and very low‐density lipoprotein were lower in fish fed LFFO than the other fish (P<0.05). The present study demonstrates that in terms of fish growth, VOs can be used as an alternate source of dietary fat, whereas fish health and nutritional value are improved with the LFFO diet. According to these results, a partial substitution of FO by VO in high‐level fat diets is suggested for long‐term feeding of Caspian brown trout.