The effect of temperature and dietary fat level on tissue lipid composition in Atlantic salmon (Salmo salar) fed wax ester‐rich oil from Calanus finmarchicus

Copepod oil (CO) from the marine zooplankton, Calanus finmarchicus, is a potential alternative to fish oils (FOs) for inclusion in aquafeeds. The oil is composed mainly of wax esters (WE) containing high levels of saturated fatty acids (SFAs) and monounsaturated fatty alcohols that are poorly digested by fish at low temperatures. Consequently, tissue lipid compositions may be adversely affected in salmon‐fed CO at low temperatures. This study examined the lipid and FA compositions of muscle and liver of Atlantic salmon reared at two temperatures (3 and 12 °C) and fed diets containing either FO or CO, supplying 50% of dietary lipid as WE, at two fat levels (～330 g kg^?1, high; ～180 g kg^?1, low). Fish were acclimatized to rearing temperature for 1 month and then fed one of four diets: high‐fat fish oil (HFFO), high‐fat Calanus oil (HFCO), low‐fat fish oil (LFFO) and low‐fat Calanus oil (LFCO). The fish were grown to produce an approximate doubling of initial weight at harvest (220 days at 3 °C and 67 days at 12 °C), and lipid content, lipid class composition and FA composition of liver and muscle were determined. The differences in tissue lipid composition between dietary groups were relatively small. The majority of FA in triacylglycerols (TAG) in both tissues were monounsaturated, and their levels were generally higher at 3 °C than 12 °C. Polyunsaturated fatty acids (PUFA), particularly (n‐3) PUFA, predominated in the polar lipids, and their level was not significantly affected by temperature. The PUFA content of TAG was highest (～26%) in the muscle of fish fed the HFCO diet at both temperatures. Tissue levels of SFAs were lower in fish‐fed diets containing HFCO than those fed HFFO, LFFO or LFCO, particularly at 3 °C. The results are consistent with Atlantic salmon being able to incorporate both the FA and fatty alcohol components of WE into tissue lipids but, overall, the effects of environmental temperature on tissue lipids were more pronounced in fish fed the CO diets than FO diets.     

Marine wax ester digestion in salmonid fish: a review

Alternative marine resources from lower trophic levels could partly cover the rapidly increasing needs for marine proteins and oils in the future. The North Atlantic calanoid copepod, Calanus finmarchicus, has a high level of lipids rich in n‐3 fatty acids. However, these animals have wax esters as the main lipid storage component rather than triacylglycerol (TAG). Although these esters are considered difficult to digest by many fish, is it well known that juvenile Atlantic salmon (Salmo salar) feed on zooplankton species. It is therefore possible that the capacity to utilize these lipids should be well developed in salmonids. Nonetheless, salmon hydrolyse wax esters slower than TAG and absorb fatty alcohols slower than fatty acids. However, salmon have several adaptations to digest diets rich in wax esters. These includes increased feed conversion, higher production of bile and higher activity of lipolytic enzymes in the midgut. Atlantic salmon has been shown to feed and grow on diets with a medium amount of wax esters (30% of the lipid) with results comparable to fish maintained on fish oil diets. Ingestion of higher level of wax esters (50% of the lipid) cause, however, poorer lipid digestibility and growth, so that optimal utilization of wax esters in Atlantic salmon is closer to 30% than 50% of the dietary lipid.     

Molecular cloning,characterization and nutritional regulation of key enzymes required for the effective utilization of marine wax esters by Atlantic salmon (Salmo salar L.)

Previous studies had shown that wax ester‐rich lipid extracted from calanoid copepods could be a useful alternative to fish oil as a provider of long‐chain n‐3 polyunsaturated fatty acids in diets for use in salmon aquaculture. Effective utilization of wax ester requires digestion and metabolism in the intestine with the fatty alcohol component being oxidized to fatty acid in intestinal cells through the combined activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). We studied wax ester utilization in Atlantic salmon using a candidate gene approach, focusing on ADH and ALDH as sequence information was available for these genes, including fish sequences, facilitating isolation of the cDNAs. Here, we report on the isolation and cloning of full‐length cDNAs for ADH3 and ALDH3a2 genes from salmon intestinal tissue. Functional characterization by heterologous expression in the yeast, Saccharomyces cerevisiae, showed the products of these cDNAs had long‐chain ADH and ALDH enzyme activities. Thus, ADH3 was capable of oxidizing long‐chain fatty alcohol, and ALDH3a2 was capable of oxidizing long‐chain fatty aldehyde to the corresponding fatty acid. The genes were highly expressed in intestinal tissue, particularly pyloric caeca, but their expression was not increased in salmon fed dietary copepod oil in comparison to fish fed fish oil.     

Effect of soy oil containing stearidonic acid on growth performance and fillet fatty acid composition of Atlantic salmon

The effects of stearidonic acid (SDA; 18:4n‐3) derived from SDA‐enhanced, genetically modified soybeans (Monsanto Company, St Louis, MO, USA) on growth performance and fatty acid (FA) composition of large Atlantic salmon (Salmo salar; 2.1 kg initial weight) were evaluated. There was a stepwise decrease in feed intake and subsequent weight gain of immature Atlantic salmon with increased replacement of fish oil by SDA soy oil from 0%, 50% to 100% added oil. SDA increased and n‐3 highly unsaturated FA (n‐3 HUFA; eicosapentaenoic acid + docosahexaenoic acid) decreased in the diet and corresponding fillet with increased SDA oil inclusion. Salmon with the same weight gain fed SDA oil compared with rapeseed oil at 50% fish oil replacement had similar n‐3 HUFA fillet levels indicating little or no increased synthesis of n‐3 HUFA from SDA for deposition in the fillet. However, elongation of dietary SDA to 20:4n‐3 for deposition in the fillet of SDA oil fed fish was indicated. The increased SDA and 20:4n‐3 in the fillet of Atlantic salmon fed SDA oil compared with rapeseed oil at 50% fish oil replacement may be more effective as precursors for EPA in humans than 18:3n‐3 which was in the fillet at similar levels.     

Effects of diets containing linseed oil on fatty acid desaturation and oxidation in hepatocytes and intestinal enterocytes in Atlantic salmon (Salmo salar)

We hypothesized that replacing fish oil with 18:3n-3-rich linseed oil may enable salmon to maintain the levels of tissue n-3HUFA levels through a combination of increased desaturation activity and increased substrate fatty acid provision. To this end we investigated desaturation/elongation of [1-¹⁴C18:3n-3 in hepatocytes and intestinal enterocytes, and determined the extent to which 18:3n-3 was oxidized and desaturated by measuring both simultaneously in a combined assay. Salmon smolts were stocked randomly into five seawater pens and fed for 40 weeks on diets in which the fish oil was replaced in a graded manner by linseed oil. At the end of the trial, fatty acyl desaturation/elongation and oxidation activities were determined in isolated hepatocytes and intestinal enterocytes using [1-¹⁴C]18:3n-3 as substrate, and samples of liver and intestinal tissue were collected for analysis of lipid and fatty acid composition. The results showed that, despite increased desaturation of [1-¹⁴C]18:3n-3 in hepatocytes, provision of dietary 18:3n-3 did not prevent the decrease in tissue n-3HUFA in fish fed linseed oil. Intestinal enterocytes were a site of significant fatty acid desaturation but, in contrast to hepatocytes, the activity was not increased by feeding linseed oil and was generally lower in fish fed linseed oil compared to fish fed only fish oil. In contrast, oxidation of [1-¹⁴C]18:3n-3 in enterocytes was generally increased in fish fed linseed oil compared to fish fed the diet containing only fish oil. However, oxidation of [1-¹⁴C]18:3n-3 in hepatocytes was 4- to 8-fold lower than in enterocytes and was not affected by diet. Furthermore, oxidation of [1-¹⁴C]18:3n-3 in enterocytes exceeded desaturation irrespective of dietary treatment, whereas similar amounts of [1-¹⁴C]18:3n-3 were desaturated and oxidized in hepatocytes from fish fed only fish oil and desaturation exceeded oxidation by 3-fold in fish fed the diet containing 100% linseed oil. The molecular mechanisms underpinning these results were discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Digestibility of Calanus finmarchicus wax esters in Atlantic salmon (Salmo salar) freshwater presmolts and seawater postsmolts maintained at constant water temperature

Calanoid copepods are a rich source of marine lipid for potential use in aquafeeds. Copepod oil is primarily composed of wax esters (WE) and there are concerns over the efficiency of wax ester, versus triacylglycerol (TAG), digestion and utilization in fish. As smoltification represents a period of major physiological adaptation, the present study examined the digestibility of a high WE diet ( Calanus oil; 48% WE, 26% TAG), compared with a TAG diet (fish oil; 58% TAG), in Atlantic salmon freshwater presmolts and seawater postsmolts, of similar age (9 months) and weight (112 g and 141 g initial, respectively), over a 98-day period at constant temperature. Fish grew significantly better, and possessed lower feed conversion ratios (FCR), in seawater than freshwater. However, total lipid apparent digestibility coefficient (ADC) values were significantly lower in seawater fish, as were total fasted bile volumes. Dietary Calanus oil also had a significant effect, reducing growth and lipid ADC values in both freshwater and seawater groups. Postsmolts fed dietary Calanus oil had the poorest lipid ADC values and analysis of faecal lipid class composition revealed that 33% of the remaining lipid was WE and 32% fatty alcohols. Dietary prevalent 22:1n-11 and 20:1n-9 fatty alcohols were particularly poorly utilized. A decrease in primary bile acid, taurocholate, concentration was observed in the bile of dietary Calanus oil groups which could be related to the lower cholesterol content of the diet. The dietary WE : TAG ratio is discussed in relation to life stage and biliary intestinal adaptation to the seawater environment postsmoltification.     

Enhancing highly unsaturated ω‐3 fatty acids in phase‐fed rainbow trout (Oncorhynchus mykiss) using Alaskan fish oils

Rainbow trout, average weight 185–187 g, were fed feeds containing menhaden oil, canola oil or fish oils (pollock, pink salmon or rockfish) produced from Alaskan seafood processing waste as the added oil for 8 weeks, at which time the fish weighed 391–411 g (average 404 g, pooled SE = 5.7). The fish were previously fed from 75 g average weight fed commercial feed containing poultry oil as the added oil. No significant differences were measured in final weight or feed conversion ratio among dietary treatment groups. Significant differences were found in fillet ω‐3 fatty acid (FA) levels from fish receiving fish oil‐supplemented feeds compared to those from fish receiving feeds containing canola oil. Fillet contents of eicosapentaenoic acid (EPA; 20:5ω3) and decosahexaenoic acid (DHA; 22:6ω3) were highest in the pollock oil treatment group, although all fish oils increased highly unsaturated ω‐3 FA contents (mg 100 g^?1) of fillets. Fish oil used through the production cycle was reduced by 25% by supplementing feeds with poultry oil during the middle phase of production (75–175 g) compared to using feeds containing fish oil throughout the production cycle. Fish oils recovered from Alaskan seafood processing waste were suitable alternatives to conventional fish oil as ingredients in rainbow trout production feeds.     

The effect of PUFA enriched Artemia on growth, survival and lipid composition of western rock lobster, Panulirus cygnus, phyllosoma

Western rock lobster, Panulirus cygnus, phyllosoma were grown from hatching to stage IV. Larvae were fed with Artemia enriched with a (i) base enrichment (Base) containing 520 g kg^?1 squid oil or tailor made enrichments in which oils high in polyunsaturated fatty acid (PUFA) have been added at the expense of squid oil. These treatments were (ii) base enrichment supplemented with docosahexaenoic acid (DHA) rich oil, (iii) base enrichment supplemented with arachidonic acid (AA) rich oil, or (iv) base enrichment supplemented with DHA and AA (D + A) rich oils. Total survival of phyllosoma to stage IV was high, with no significant difference between treatments (range 12.3–17.5%). By stage IV, the larvae fed the DHA or AA enriched Artemia were significantly larger (3.33 mm length) than larvae fed the Base or D + A enriched Artemia (3.18–3.24 mm length). Phyllosoma were sampled at stages II and III for biochemical analysis. The major lipid class (LC) in all phyllosoma was polar lipid (PL) (88.9–92.4%), followed by sterol (ST) (6.2–9.7%). Triacylglycerol (TAG), free fatty acid (FFA) and hydrocarbon/wax ester were minor components (≤1%) in all phyllosoma samples. In contrast, the major LC in all enrichments and enriched Artemia was TAG (76.3–85.1% and 53.4–60.2%, respectively), followed by PL (11.4–14.8% and 30.6–38.1% respectively). The main fatty acids (FA) in phyllosoma were 16:0, 18:1n‐9, 18:1n‐7, 18:0, AA, eicosapentaenoic acid (EPA) and DHA. Addition of AA, and to a lesser extent DHA, to enrichments resulted in increased levels of those FA in Artemia and phyllosoma compared with the Base enrichment. This was particularly evident for stage III larvae. Comparatively, elevated growth for phyllosoma to stage IV was achieved with DHA and AA enriched diets. Our findings highlight the importance of lipids and in particular essential long‐chain PUFA, as nutritional components for phyllosoma diets.     

Complete replacement of dietary fish oil with a vegetable oil blend affect liver lipid and plasma lipoprotein levels in Atlantic salmon (Salmo salar L.)

To study how hepatic lipid turnover and lipid transport may be affected by complete replacement of dietary fish oil (FO) with a vegetable oil blend (VO) from start feeding until the adult stages, Atlantic salmon (Salmo salar L.) were fed either 100% FO‐ or 100% VO‐based diets (55% rapeseed oil, 30% palm oil and 15% linseed oil) from start feeding until 22 months. Liver and plasma lipoprotein lipid class levels and lipoprotein fatty acid composition were analysed through the seawater phase, whereas liver fatty acid composition, plasma cholesterol, triacylglycerol (TAG) and protein levels were analysed through both freshwater and seawater stages. Further, enzyme activity of liver fatty acid synthetase (FAS), NADH‐isocitrate dehydrogenase, malic enzyme, glucose‐6‐phosphate dehydrogenase and 6‐phosphogluconate dehydrogenase and expression of the gene Peroxisome proliferator‐activated receptor γ (PPARγ) was analysed during both fresh water and seawater stages through the experiment. Dietary VO significantly increased salmon liver TAG and hence total liver lipid stores after 14 and 22 months of feeding. Further, after 22 months of feeding, plasma lipid levels and plasma low‐density lipoprotein (LDL) levels were significantly decreased in VO‐fed salmon compared with FO‐fed fish. The same trend, although not statistically significant, was seen for plasma very low‐density lipoprotein (VLDL). The activity of FAS was generally low throughout the experiment with the VO group having significantly lower activity after 16 months of feeding. The expression of PPARγ in livers increased prior to seawater transfer followed by a decrease, and then another increase towards the final sampling (22 months). Dietary vegetable oil replacement had no impact on PPARγ expression in salmon liver. In summary, liver TAG stores, plasma lipid and LDL levels were affected by dietary vegetable oil replacement in Atlantic salmon during a long–term feeding experiment. Current results indicate that high dietary vegetable oil inclusion increase hepatic TAG stores and decrease plasma lipid levels possible through decreased VLDL synthesis.     

Nutritional and environmental regulation of the synthesis of highly unsaturated fatty acids and of fatty-acid oxidation in Atlantic salmon (Salmo salar L.) enterocytes and hepatocytes

The objective of this work was to determine whether highly unsaturated fatty acid (HUFA) synthesis and fatty-acid oxidation in Atlantic salmon (Salmo salar L.) intestine was under environmental and/or seasonal regulation. Triplicate groups of salmon were grown through a full two-year cycle on two diets containing either fish oil (FO) or a diet with 75% of the FO replaced by a vegetable oil (VO) blend containing rapeseed, palm, and linseed oils. At key points in the life cycle fatty acyl desaturation/elongation (HUFA synthesis) and oxidation activity were determined in enterocytes and hepatocytes using [1−¹⁴C]18:3n−3 as substrate. As observed previously, HUFA synthesis in hepatocytes reached a peak at seawater transfer and declined thereafter, with activity consistently greater in fish fed the VO diet. In fish fed FO, HUFA synthesis in enterocytes in the freshwater stage was at a level similar to that in hepatocytes. HUFA synthesis in enterocytes increased rapidly after seawater transfer, however, and remained high for some months after transfer before decreasing to levels that were again similar to those observed in hepatocytes. Enterocyte synthesis of HUFA was usually higher in fish fed the VO diet than in those fed the FO diet. Oxidation of [1−¹⁴C]18:3n−3 in hepatocytes from fish fed FO tended to decrease during the freshwater phase but then increased steeply, peaking just after transfer before decreasing during the remaining seawater phase. At the peak in oxidation activity around seawater transfer, activity was significantly lower in fish fed VO than in fish fed FO. In enterocytes, oxidation of [1−¹⁴C]18:3 in fish fed FO reached a peak in activity just before seawater transfer. In fish fed VO, except for high activity at nine months the pattern was similar to that obtained in enterocytes from fish fed FO, with high activity around seawater transfer and declining activity in seawater. In conclusion, fatty acid metabolism in intestinal cells seemed to be under dual nutritional and environmental or seasonal regulation. Temporal patterns of oxidation of fatty acids were usually similar in the two cell types, but HUFA synthesis in enterocytes peaked over the summer seawater phase rather than at transfer, as with hepatocytes, suggesting the possibility of different regulatory cues.     

Influence of dietary palm oil on growth, tissue fatty acid compositions, and fatty acid metabolism in liver and intestine in rainbow trout (Oncorhynchus mykiss)

This study aimed to investigate the effects of dietary crude palm oil (CPO) on fatty acid metabolism in liver and intestine of rainbow trout. Triplicate groups of rainbow trout for 10 weeks at 13 °C were fed on diets in which CPO replaced fish oil (FO) in a graded manner (0–100%). At the end of the trial, fatty acid compositions of flesh, liver and pyloric caeca were determined and highly unsaturated fatty acid (HUFA) synthesis and fatty acid oxidation were estimated in isolated hepatocytes and caecal enterocytes using [1‐¹⁴C]18:3n‐3 as substrate. Growth performance and feed efficiency were unaffected by dietary CPO. Fatty acid compositions of selected tissues reflected the dietary fatty acid composition with increasing CPO resulting in increased proportions of 18:1n‐9 and 18:2n‐6 and decreased proportions of n‐3HUFA, 20:5n‐3 and 22:6n‐3. Palmitic acid, 16:0, was also increased in flesh and pyloric caeca, but not in liver. The capacity of HUFA synthesis from 18:3n‐3 increased by up to threefold in both hepatocytes and enterocytes in response to graded increases in dietary CPO. In contrast, oxidation of 18:3n‐3 was unaffected by dietary CPO in hepatocytes and reduced by high levels of dietary CPO in enterocytes. The results of this study suggest that CPO can be used at least to partially replace FO in diets for rainbow trout in terms of permitting similar growth and feed conversion, and having no major detrimental effects on lipid and fatty acid metabolism, although flesh fatty acid compositions are significantly affected at an inclusion level above 50%, with n‐3HUFA reduced by up to 40%.     

Effects of oxidized fish oil intake on tissue lipid metabolism and fatty acid composition of channel catfish (Ictalurus punctatus)

Recent studies in terrestrial animals have shown that feeding the oxidized lipids led to a reduction in triacylglycerols (TAG) and total cholesterol (TC) in liver and plasma. However, limited information is available on the effect of oxidized lipids on lipid metabolism in fish. In this study, four diets containing 0 g kg^?1 (control: fresh fish oil), 30 g kg^?1 (low‐oxidized oil, LOO), 60 g kg^?1 (medium‐oxidized oil, MOO) and 90 g kg^?1 (high‐oxidized oil, HOO) graded oxidized oil levels with the same dietary lipid level were fed to channel catfish for 86 days. The tissue lipid metabolism and fatty acid composition of the fish were investigated after this period. The results showed that plasma and liver concentrations of TAG and TC decreased with increasing dietary oxidized oil level (P < 0.05). Decreasing liver lipoprotein lipase and hepatic lipase activities were observed with increasing dietary oxidized fish oil inclusion (P < 0.05). The liver C22:6n?3 concentrations significantly decreased with increasing dietary oxidized oil level (P < 0.05), while muscle lipid had a high proportion of polyunsaturated fatty acids. It suggests that the adverse effects of dietary oxidized oil may be induced by inhibiting lipid metabolism enzymes and, consequently, inhibition of cholesterol homoeostasis and fatty acid synthesis.     

Tissue and organ distribution of 14C‐activity in dextrin‐adapted Atlantic salmon after oral administration of radiolabelled 14C1‐glucose

Accumulation of ¹⁴C in various tissues and organs was studied in three different groups of 0.8‐kg Atlantic salmon Salmo salar force‐fed with ¹⁴C₁‐glucose in order to evaluate if metabolism of glucose depended on adaptation to dietary carbohydrate level. The salmon had been fed diets supplemented with 0, 100 and 200 g maize dextrin kg^?1 for 10 months before the experiment. The fish were force‐fed 6.65 × 10⁴ Bq of ¹⁴C₁ glucose kg^?1 BW, in gelatin capsules. Fish for analysis were obtained 16 h later. ¹⁴C was measured in blood plasma, gill, kidney, liver and white muscle, and in lipid extract of liver. The liver contained most ¹⁴C, followed by heart, blood plasma, gill and liver lipid extract, while kidney and muscle contained the least ¹⁴C per gram or millilitre tissue. The muscle contained most radioactivity, on an estimated total tissue basis, followed by liver, blood plasma, gill, liver lipid extract, kidney and heart tissue. Thirty‐eight per cent of the orally administered ¹⁴C was recovered in the salmon adapted to the diet without dextrin after 16 h. This was significantly (P < 0.05) higher than the 30% and 32% recovered in the salmon adapted to diets with 10% and 20% dextrin. This effect on adaptation to dietary dextrin level in glucose uptake or metabolism was supported by a trend (P < 0.10) toward higher radioactivity per gram or millilitre of each individual tissue in the fish adapted to the diet without dextrin, when compared with the other two adaptation regimes.     

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.     

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.     

Alternative vegetable lipid sources in diets for grouper, Epinephelus coioides (Hamilton): effects on growth, and muscle and liver fatty acid composition

The aim of this study was to investigate the effects of different oils on growth performance and lipid metabolism of the grouper, Epinephelus coioides. Five experimental fish meal‐based isonitrogenous and isolipidic diets were formulated containing either 5.5%‐added fish oil (FO), soybean oil (SBO), corn oil (CO), sunflower oil (SFO) or peanut oil (PO). Each diet was fed to triplicate groups of 20 fish (initial body weight 13.2±0.02 g) grown in seawater at 28.0–30.5 °C for 8 weeks. Fish were fed twice a day to visual satiety. No significant differences in the survival, weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio or hepatosomatic index were found between fish fed the FO or vegetable oils (VO) diets. Dietary lipid sources did not affect whole‐body composition among grouper fed the various diets. Muscle of fish fed the FO diet had significantly higher levels of 14:0, 16:0, 16:1n‐7, 20:5n‐3[eicosapentaenoic acid (EPA)] and docosahexaenoic acid (DHA)+EPA (except for PO fed fish) compared with those of fish fed VO diets. However, the levels of 18:1n‐9, 18:2n‐6 and DHA/EPA ratios in the muscle of fish fed FO diet were significantly lower than those of fish fed the VO diets. The liver of fish fed the FO diet had significantly higher levels of 18:0, 20:5n‐3, 22:6n‐3, n‐3 highly unsaturated fatty acids and DHA+EPA than those of fish fed the VO diets, whereas increases in 18:1n‐9, 18:2n‐6 and mono‐unsaturated fatty acid levels were observed in the liver of fish fed the VO diets.     

Effects of varying dietary fatty acid profile on growth performance, fatty acid, body and tissue composition of juvenile pike perch (Sander lucioperca)

Pike perch (Sander lucioperca) has been identified as specie destined to diverse European inland aquaculture, but knowledge on the nutritional requirements is weak. Therefore, we investigated the effect of varying dietary fatty acid (FA) profile by partial replacement of fish oil (FO) with vegetable oils on growth, FA and body composition of juvenile pike perch. An extruded basal diet containing 59 g kg^?1 crude lipids (FO) was added with 60 g kg^?1 FO, 60 g kg^?1 linseed oil (LO) or 60 g kg^?1 soybean oil (SO). The resulting dietary FA composition differed mainly in the triglyceride fraction and was characterized by highest amounts of linolenic acid (18:3 n‐3) in the LO diet and linoleic acid in the SO diet. Diet enriched with FO contained highest contents of highly unsaturated FA 20:5 n‐3 (eicosapentaenic acid) and 22:6 n‐3 (docosahexaenic acid). Pike perch were held in a recirculation system and each feeding group (in triplicate) was fed with experimental diets at a daily rate of 35 g kg^?1 of biomass for 57 days by automatic feeders. Weight gain and specific growth rate of experimental feeding groups ranged between 18.47 and 19.58 g and 1.37–1.45% day^?1 and was not affected by the dietary composition indicating that FO can be replaced by vegetable oils without negative impact on growth performance. In contrast to the whole body and muscle composition, liver tissue was affected by the varying diets. Liver tissues of fish fed diets enriched with vegetable oils showed significantly increased lipid contents of 162 (LO) and 147 (SO) g kg^?1 and indicate decreased lipid utilization compared with fish fed FO diet (liver lipid content 112 g kg^?1). Nevertheless, hepatosomatic index of pike perch was not influenced by dietary lipid composition. The FA profile of pike perch was generally determined by the dietary FAs.     

Atlantic salmon, Salmo salar, utilizes wax ester-rich oil from Calanus finmarchicus effectively

Against a background of decreasing availability of fish oils for use in aquaculture, the present study was undertaken to examine whether a wax ester-rich oil derived from the calanoid copepod Calanus finmarchicus, could be used effectively by Atlantic salmon when supplied in their diet. Individually tagged Atlantic salmon of initial weight around 500 g were divided into replicate tanks of two dietary groups and fed either a fish oil supplemented diet, or an experimental diet coated with Calanus oil. Wax esters accounted for 37.5% of the lipids in the Calanus oil diet but were absent from the fish oil diet in which triacylglycerols (TAG) were the major lipid class. Over the feeding period (140 days) the salmon fed fish oil displayed a greater increase in length, but there was no significant difference between the two groups in weight gained. The specific growth rates (0.75) and the feed conversion ratio of fish fed the two diets were similar throughout the study. No differences were observed in the apparent digestibility coefficients (ADC) of fish fed Calanus oil or fish oil. The ADC of fatty acids decreased with chain length and increased with unsaturation. Long-chain alcohol utilization showed a similar tendency although there was a notable difference in that saturated long-chain alcohols were utilized better than the comparable fatty acid homologue. In fecal lipid of fish fed Calanus oil, the content of 16:0 alcohol decreased in both the free long-chain alcohol and wax ester fractions, while the corresponding fatty acid increased in the feces of both dietary groups of fish. In contrast, the proportion of the 22:1n−11 alcohol increased in both fecal wax esters and free long-chain alcohol fractions whereas 22:1n−11 fatty acid displayed no accumulation. The observed patterns of fatty acid and long-chain alcohol compositions in fecal lipid compared to those of the initial dietary lipid are consistent with the digestive lipases of salmon preferentially hydrolyzing esters containing polyunsaturated fatty acid (PUFA) moieties. The wax esters of Calanus oil contained substantial amounts of the n−3 PUFA, 20:5n−3 and 22:6n−3, that were effectively deposited in muscle and liver tissues. No major differences were seen in either lipid content/lipid classes or in gross fatty acid composition of these tissues between the two dietary groups. It is concluded that that Atlantic salmon in seawater can effectively utilize diets in which a major lipid component is derived from zooplankton rich in wax ester without any detrimental change in growth or body lipid composition. This finding gives support to the use of lipid from zooplankton from high latitudes as an alternative or as a supplement to fish oil and a provider of long-chain n−3 PUFA in diets for use in salmon aquaculture.     

Efficacy of various lipid supplements in formulated pellet diets for juvenile Scylla serrata

Efficacy of sunflower oil (diet SF) and soybean oil (diet SB) alone and in combination with cod liver oil (diets M1‐2.80:1.40:1.40, M2‐2.80:2.24:0.56 and M3‐2.80:0.56:2.24; cod liver oil:sunflower oil:soybean oil) as lipid supplements (5.6%) in formulated diets (crude fat ～9.79%) for juvenile Scylla serrata (weight=0.28±0.07 g, carapace width=9.7±0.1 mm) were compared with diet CL, containing cod liver oil alone as the lipid supplement (6 diets × 24 crabs stocked individually, randomized block design). Growth performance, nutrient (protein and lipid) intake and gain of crabs fed M1, M2 and M3 were higher (P≤0.05) than the crabs fed SF and SB, but were not significantly different (P≥0.05) from crabs fed CL. Dietary fatty acids (FAs) are found to influence the FA profile of test crabs. Higher tissue levels of 16:1n‐7, 18:1n‐9 and 18:1n‐7 reflected the essential FA deficiency in crabs fed diets supplemented only with vegetable oils. Results confirmed that S. serrata could utilize vegetable oil supplements in the formulated diets as a partial replacement (50%) of cod liver oil without compromising growth and survival. Partial substitution of marine fish oil with suitable vegetable oils can reduce the feed cost considerably, in the context of rising fish oil prices.     

The uptake and esterification of radiolabelled fatty acids by enterocytes isolated from rainbow trout (Oncorhynchus mykiss)

To study the intestinal fatty acid absorption in fish in vitro, enterocytes were isolated from the intestine of rainbow trout and incubated with an equimolar mixture of seven fatty acids [16:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3] in which the component carrying a radioactive label was varied. The fatty acid mixture was presented in the form of micelles formed by sonication with sodium taurocholate. Control studies showed that the presence of sodium taurocholate in the incubation medium caused an immediate 23% increase in the mortality of the cells but the remaining cells were viable. The effect of the bile salt on cellular permeability was evident at longer exposure periods. The proportions of 14C-labelled 20:5n-3 and 22:6n-3 present as monomeric fatty acids in the micellar solutions were higher than those of 16:0, 18:1n-9, 18:2n-6, 18:3n-3 and 20:4n-6. The rates of uptake of 14C-labelled 20:4n-6, 20:5n-3 and 22:6n-3 by enterocytes were significantly lower than those of the other fatty acid substrates over the first minute of incubation but no significant differences in uptake rate between fatty acids were obvious over a 15 min incubation period. Notable differences were observed between substrates in their distribution pattern in enterocyte lipid classes. Although most of the radioactivity from all radiolabelled substrates was recovered in triacylglycerols, the amounts of 14C- labelled 16:0, 20:4n-6, 20:5n-3 and 22:6n-3 recovered in the polar lipid fraction were higher than those of 14C-labelled C18 substrates, particularly after 15 min. Conversely, the initial esterification rates of 18:1n-9, 18:2n-6 and 18:3n-3 into triacylglycerols were significantly higher than those of 20:4n-6, 20:5n-3 and 22:6n-3. It is concluded that isolated enterocytes can be used for the study of the mechanism of intestinal fatty acid absorption in fish.