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Meagre (Argyrosomus regius) is a promising new aquaculture species, however with some bottlenecks to high-scale commercial production. One of those constraints are the scarce information about its nutritional requirements, which difficult the production of optimized diets and can lead to pathological alterations in liver, such as hepatic steatosis. Furthermore, meagre production is widely affected by granulomatosis, an infectious or non-infectious disease that may compromise fish performance and appearance. The cause of non-infectious granulomatosis is still unclear, but nutritional deficiencies are potential explanations. Since essential fatty acids are nutrients required to sustain vital functions, growth, but also immune status and disease resistance, the present work evaluated, in a 30-day feeding trial, the effect of 5 dietary n-3 long-chain polyunsaturated fatty acid levels (0.8, 1.4, 2.0, 2.6 and 3.5% in dry weight of the diet) on liver health status of meagre fingerlings. Fish fed the lowest n-3 LC-PUFA diets (0.8 and 1.4%) showed a higher hepatosomatic index, as well as a higher hepatic lipid infiltration, which resulted in severe hepatic steatosis. Oleic acid and docosahexaenoic acid were the most important fatty acids driving the variability of meagre hepatic fatty acid composition and, consequently, hepatic histological alterations. Although fatty acid composition of phospholipids was less affected by the dietary n-3 LC-PUFA, meagre fed 0.8% n-3 LC-PUFA tended to present lower levels of essential fatty acids and higher levels of oleic acid in polar lipids. Moreover, the diet with the lowest n-3 LC-PUFA content (0.8%) led to a higher incidence of hepatic granulomas, suggesting a possible relation between essential fatty acid deficiency and hepatic granulomatosis in meagre. These results suggest that the inclusion of vegetable oils in diets for meagre fingerlings should be taken with careful, guaranteeing that, at least, 2.0% DW of n-3 LC-PUFA is available in feeds, in order to maintain the normal hepatic function in meagre.

Earlier studies have indicated that a high inclusion of n-6 fatty acids (FA) in feeds for Atlantic salmon can affect the stress response. To test this hypothesis, Atlantic salmon (Salmo salar) were fed diets containing varying dietary n-6/n-3 FA ratios and different absolute levels of n-6 and n-3 FAs. The fish were divided into two different stress challenge groups, where one group was exposed to three weekly hypoxia challenges for 4 weeks (repeated stress), while one group was left undisturbed. At the end of the experiment, both groups were exposed to an acute stressor (lowering of water level). Thus, effects of the diets on acute stress, repeated stress and the combined effect of these could be investigated. In general, there were few effects of the repeated stress, while fish in all diet groups responded strongly to the acute stress based on several stress markers. Dietary n-6/n-3 ratio did not affect growth, all fish appeared phenotypically healthy, and all groups were able to mount an acute stress response. However, there was an interaction between diet and repeated stress on cortisol response after acute stress, possibly indicating altered hypothalamic-pituitary adrenal axis reactivity in fish fed high n-6/n-3 FA ratio. Hepatic levels of prostaglandin D2 (PGD2) and leukotriene B4 responded differently to acute stress depending on the dietary n-6/n3 FA ratio, indicating an altered acute stress response. Additionally, increasing the dietary n-6/n-3 FA content led to higher levels of PGD2 and PGE2 as well as higher liver triacylglycerol. In summary, the results suggest that increasing the dietary n-6/n-3 FA ratio in salmon feeds can affect the way they respond to stressors in an aquaculture setting, possibly affecting the fish robustness.

The substitution of fish oil and fishmeal with plant-based ingredients in commercial aquafeeds for Atlantic salmon, may introduce novel contaminants that have not previously been associated with farmed fish. The organophosphate pesticide pirimiphos-methyl (PM) is one of the novel contaminants that is most prevalent in commercial salmon feed. In this study, the feed-to-fillet transfer of dietary PM and its main metabolites was investigated in Atlantic salmon fillet. Based on the experimental determined PM and metabolite uptake, metabolisation, and elimination kinetics, a physiologically based toxicokinetic (PBTK) compartmental model was developed. Fish fed PM had a relatively low (~4%) PM retention and two main metabolites (2-DAMP and Desethyl-PM) were identified in liver, muscle, kidney and bile. The absence of more metabolised forms of 2-DAMP and Desethyl-PM in Atlantic salmon indicates different metabolism in cold-water fish compared to previous studies on ruminants. The model was used to simulate the long term (>1.5 years) feed-to-fillet transfer of PM + metabolite in Atlantic salmon under realistic farming conditions including seasonal fluctuations in feed intake, growth, and fat deposition in muscle tissue. The model predictions show that with the constant presence of the highest observed PM concentration in commercial salmon feed, fillet PM+ metabolite levels were approximately 5 nmol kg−1, with highest levels for the metabolite 2-DAMP. No EU maximum residue levels (MRL) for PM and its main metabolites exist in seafood to date, but the predicted levels were lower than the MRL for PM in swine of 32.7 nmol kg−1.

PurposeTo explore whether high intake of cod or salmon would affect gut microbiota profile, faecal output and serum concentrations of lipids and bile acids.MethodsSeventy-six adults with overweight/obesity with no reported gastrointestinal disease were randomly assigned to consume 750 g/week of either cod or salmon, or to avoid fish intake (Control group) for 8 weeks. Fifteen participants from each group were randomly selected for 72 h faeces collection at baseline and end point for gut microbiota profile analyses using 54 bacterial DNA probes. Food intake was registered, and fasting serum and morning urine were collected at baseline and end point.ResultsSixty-five participants were included in serum and urine analyses, and gut microbiota profile was analysed for 33 participants. Principal component analysis of gut microbiota showed an almost complete separation of the Salmon group from the Control group, with lower counts for bacteria in theBacteroidetesphylum and theClostridialesorder of theFirmicutesphyla, and higher counts for bacteria in theSelenomonadalesorder of theFirmicutesphylum. The Cod group showed greater similarity to the Salmon group than to the Control group. Intake of fibres, proteins, fats and carbohydrates, faecal daily mass and output of fat, cholesterol and total bile acids, and serum concentrations of cholesterol, triacylglycerols, non-esterified fatty acids and total bile acids were not altered in the experimental groups.ConclusionA high intake of cod or salmon fillet modulated gut microbiota but did not affect faecal output or serum concentrations of lipids and total bile acids.Clinical trial registrationThis trial was registered at clinicaltrials.gov as NCT02350595.

A well-balanced diet adapted to fulfil the specific nutritional requirements of greater amberjack (Seriola dumerili) would contribute to optimize reproduction and spawning quality. The main objective of the present study was to examine the effects of supplemental histidine, taurine and protein in broodstock diets, on egg quality of greater amberjack. Twelve broodstocks were distributed in three 40 m3 circular tanks and fed three different diets, one high in histidine, another in taurine and the third one in protein contents. Overall the best spawning quality was obtained histidine levels were elevated from 1 to 1.5%, increasing the relative fecundity, fertilization rates, egg viability, hatching rates, larval survival, egg protein contents, and egg and larval size, denoting the importance of this amino acid for embryo and larval development. The increase of taurine from 0.3 to 1.1% in diets for greater amberjack increased the relative fecundity in comparison to fish fed higher protein levels. Fertilization rates tend to increase with the elevation of dietary taurine and were not significantly different from those of broodstock fed higher histidine levels. Besides, dietary taurine increased egg diameter and taurine contents. Increase in dietary protein contents from 51 to 56% lead to an increase protein content in egg, as well as a larger yolk sac volume, but did not improve any of the spawning quality parameters. In conclusion, the results of this study have pointed out the importance of raising histidine contents in broodstock diets from 1 to 1.5% to optimize the reproductive performance of greater amberjack. Besides, the study showed that taurine levels in broodstock diets increased fecundity, maintaining good fertilization rates, but further studies must be conducted to determine the optimum taurine dietary levels.

Systemic granulomatosis is a growing disease with a high morbidity, which affects to the majority of farmed meagre (Argyrosomus regius). The impossibility of isolating any infectious agents has hypothesized a nutritional origin of the disease. In order to try to elucidate the nutritional origin of granulomas, juvenile meagre were fed for 15 weeks with six diets containing different levels of vitamin E and C and with or without addition of vitamin K: Diet 0 (basal premix, no K, 150 mg kg‐−1 E, 20 mg kg‐−1 C), K (added 23 mg kg‐−1 K), EC (300 mg kg‐−1 E, 70 mg kg‐−1 C), KEC (23 mg kg‐−1 K, 300 mg kg‐−1 E, 70 mg kg‐−1 C), EECC (450 mg kg‐−1 E, 230 mg kg‐−1 C) and KEECC (23 mg kg‐−1 K, 450 mg kg‐−1 E, 230 mg kg‐−1 C). The diet EC significantly increased meagre growth in terms of final weight and length. Fish fed the highest levels of vitamin E and C presented lower percentage of granulomas in liver and heart than fish fed diet 0. The scored severity of granulomatosis in liver and kidney (main affected organs) tended to be lower with dietary increase of vitamin E, C and addition of vitamin K (from 1.83 diet 0 to 1.3 diet KEECC and from 0.91 diet 0 to 0.39 diet KEECC). In liver, the diet KEECC significantly increased catalase expression compared with diet 0. In kidney tnfα expression was significantly up-regulated in fish fed diet EECC and KEECC. In heart, low vitamin E and C levels (300 and 70 mg kg‐−1, respectively) significantly increased superoxide dismutase and glutathione peroxidase expression and high addition increased the expression of tnfα and cox-2 (0 or 23 mg kg‐−1 K, 450 mg kg‐−1 E and 230 mg kg‐−1 C, diet EECC and KEECC). The results show that combination of high dietary content of vitamin K and antioxidant vitamins E and C (23, 450 and 230 mg kg‐−1, respectively) influenced in the incidence of the granulomatosis, which suggests that this pathology could be mediated by nutritional factors.

The aim of the study was to investigate how the dietary sterol composition, including cholesterol, phytosterol:cholesterol ratio and phytosterols, affect the absorption, biliary excretion, retention, tissue storage and distribution of cholesterol and individual phytosterols in Atlantic salmon (Salmo salarL.). A feeding trial was conducted at two different temperatures (6 and 12°C), using nine different diets with varying contents of phytosterols, cholesterol and phytosterol:cholesterol ratio. Cholesterol retention values were clearly dependent on dietary cholesterol, and showed that fish fed cholesterol levels de novo. Despite this production, cholesterol content increased with increasing dietary cholesterol in liver, plasma, bile, muscle, adipose tissue and whole fish at 12°C, and in plasma, bile and whole fish at 6°C. The tissue sterol composition generally depended on the dietary cholesterol content and on the dietary phytosterol:cholesterol ratio, but not on the dietary phytosterol content in itself. Campesterol and brassicasterol appeared to be the phytosterols with the highest intestinal absorption in Atlantic salmon. There was a high biliary excretion of campesterol, but not of brassicasterol, which accumulated in tissues and particularly in adipose tissue, with 2-fold-higher retention at 12°C compared with 6°C. Campesterol had the second highest retention of the phytosterols in the fish, but with no difference between the two temperatures. Other phytosterols had very low retention. Although brassicasterol retention decreased with increasing dietary phytosterols, campesterol retention decreased with increasing dietary cholesterol, indicating differences in the uptake mechanisms for these two sterols.

The increasing use of plant-based ingredients in aquafeeds for Atlantic salmon has led to increasing feed concentrations of plant sterols (phytosterols) combined with decreasing concentrations of cholesterol. Questions have been raised regarding how this may affect performance, health and lipid metabolism in a carnivorous fish species adapted to a high cholesterol diet in nature, and whether phytosterols could be involved in the liver lipid accumulation commonly observed in salmon fed diets with high content of vegetable oils. A feeding trial with 9 diet groups with varying additions of phytosterols and cholesterol was conducted at two temperatures (6 and 12 °C). Fish grew from an initial weight of ~ 250 g to a final weight of ~ 900 g. Overall, the addition of phytosterols and/or cholesterol within the range tested (1353–3147 mg kg− 1 phytosterols, 867–3153 mg kg− 1 cholesterol and phytosterol:cholesterol ratios of 0.7–3.2) exhibited few effects on fish performance, lipid metabolism and health. The main hypothesis regarding phytosterols or phytosterol:cholesterol ratio affecting liver lipid accumulation appears to be false. However, dietary cholesterol did increase muscle pigmentation at both temperatures, while fillet dry matter and hepatosomatic index were also increased by dietary cholesterol at 12 °C. Dietary cholesterol also had some minor effects on the fatty acid composition of liver polar lipids, probably mediated by LXR. In general, more effects were seen from variations in dietary cholesterol compared to phytosterols. The current trial is the first to feed salmon graded levels of phytosterols, cholesterol and their ratio without changing the dietary fatty acid composition, and thereby isolating the effects of dietary sterols.

To identify biomarkers to assess participants’ compliance in an intervention study with high intake of cod or salmon, compared to a fish-free diet. In this randomised clinical trial, 62 healthy overweight/obese participants consumed 750 g/week of either cod (N = 21) or salmon (N = 22) across 5 weekly dinners, or were instructed to continue their normal eating habits but avoid fish intake (Control group, N = 19) for 8 weeks. After cod intake, serum concentrations of trimethylamine N-oxide (TMAO, p = 0.0043), creatine (p = 0.024) and 1-methylhistidine (1-MeHis, p = 0.014), and urine concentrations (relative to creatinine) of TMAO (p = 2.8 × 10−5), creatine (p = 8.3 × 10−4) and 1-MeHis (p = 0.016) were increased when compared to Control group. After salmon intake, serum concentrations of 1-MeHis (p = 2.0 × 10−6) and creatine (p = 6.1 × 10−4), and urine concentrations (relative to creatinine) of 1-MeHis (p = 4.2 × 10−6) and creatine (p = 4.0 × 10−5) were increased when compared to Control group. Serum and urine concentrations of TMAO were more increased following cod intake compared to salmon intake (p = 0.028 and 2.9 × 10−4, respectively), and serum and urine 1-MeHis concentrations were more increased after salmon intake compared to cod intake (p = 8.7 × 10−5 and 1.2 × 10−4, respectively). Cod and salmon intake did not affect serum and urine concentrations of 3-methylhistidine, and only marginally affected concentrations of free amino acids and amino acid metabolites. TMAO measured in serum or urine is a potential biomarker of cod intake, and 1-MeHis measured in serum or urine is a potential biomarker of salmon intake.

// Ingvild Eide Graff 1 , Jannike Oyen 1 , Marian Kjellevold 1 , Livar Froyland 1 , Clara Gram Gjesdal 2,3 , Bjorg Almas 4 , Grethe Rosenlund 5 and Oyvind Lie 1 1 National Institute of Nutrition and Seafood Research (NIFES), Bergen, Norway 2 Department of Rheumatology, Haukeland University Hospital, Bergen, Norway 3 Department of Clinical Science, University of Bergen, Bergen, Norway 4 Hormone Laboratory, Haukeland University Hospital, Bergen, Norway 5 Skretting Aquaculture Research Centre, Stavanger, Norway Correspondence to: Jannike Oyen, email: // Keywords : Atlantic salmon, bone health, vitamin D, vitamin K, bone biomarkers, Gerotarget Received : March 09, 2016 Accepted : June 09, 2016 Published : August 14, 2016 Abstract Suboptimal vitamin D status is common among humans, and might increase bone resorption with subsequent negative effects on bone health. Fatty fish, including Atlantic salmon, is an important dietary vitamin D source. However, due to a considerable change in fish feed composition, the contribution of vitamin D from salmon fillet has been reduced. The main objective was to investigate if intake of vitamin D 3 enriched salmon or vitamin D 3 tablets decreased bone biomarkers (urinary N-telopeptides, deoxypyridinoline, serum bone-specific alkaline phosphatase, and osteocalcin) compared to a low vitamin D 3 intake. The 122 healthy postmenopausal women included in this 12 weeks intervention trial were randomized into four groups: three salmon groups (150 grams/two times/week) and one tablet group (800 IU vitamin D and 1000 mg calcium/day). The salmon groups also received calcium supplements. The salmon had three different vitamin D 3 /vitamin K 1 combinations: high D 3 +high K 1 , low D 3 +high K 1 , or high D 3 +low K 1 . Increased intake of salmon containing high levels of vitamin D 3 (0.35-0.38 mg/kg/fillet) and supplements with the same weekly contribution had a positive influence on bone health as measured by bone biomarkers in postmenopausal women. Consequently, an increased level of vitamin D 3 at least to original level in feed for salmonids will contribute to an improved vitamin D 3 status and may improve human bone health.

The aim of the present study was to examine whether high intake of lean or fatty fish (cod and farmed salmon, respectively) by healthy, normal-weight adults would affect risk factors of type 2 diabetes and CVD when compared with lean meat (chicken). More knowledge is needed concerning the potential health effects of high fish intake (>300 g/week) in normal-weight adults. In this randomised clinical trial, thirty-eight young, healthy, normal-weight participants consumed 750 g/week of lean or fatty fish or lean meat (as control) for 4 weeks at dinner according to provided recipes to ensure similar ways of preparations and choices of side dishes between the groups. Energy and macronutrient intakes at baseline and end point were similar in all groups, and there were no changes in energy and macronutrient intakes within any of the groups during the course of the study. High intake of fatty fish, but not lean fish, significantly reduced TAG and increased HDL-cholesterol concentrations in fasting serum when compared with lean meat intake. When compared with lean fish intake, fatty fish intake increased serum HDL-cholesterol. No differences were observed between lean fish, fatty fish and lean meat groups regarding fasting and postprandial glucose regulation. These findings suggest that high intake of fatty fish, but not of lean fish, could beneficially affect serum concentrations of TAG and HDL-cholesterol, which are CVD risk factors, in healthy, normal-weight adults, when compared with high intake of lean meat.

The present study was conducted to evaluate the effects on Atlantic salmon hepatic lipid metabolism when fed diets with increasing substitution of fish oil (FO) with a vegetable oil (VO) blend. Four diets with VOs replacing 100, 90, 79 and 65 % of the FO were fed for 5 months. The levels of eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) in the experimental diets ranged from 1.3 to 7.4 % of fatty acids (FAs), while cholesterol levels ranged from 0.6 to 1.2 g kg(-1). In hepatocytes added [1-(14)C] α-linolenic acid (ALA, 18:3n-3), more ALA was desaturated and elongated to EPA and DHA in cells from fish fed 100 % VO, while in fish fed 65 % VO, ALA was elongated to eicosatrienoic acid (ETE; 20:3n-3), indicating reduced Δ6 desaturation activity. Despite increased desaturation activity and activation of the transcription factor Sp1 in fish fed 100 % VO, liver phospholipids contained less EPA and DHA compared with the 65 % VO group. The cholesterol levels in the liver of the 100 % VO group exceeded the levels in fish fed the 65 % VO diet, showing an inverse relationship between cholesterol intake and liver cholesterol content. For the phytosterols, levels in liver were generally low. The area as a proxy of volume of lipid droplets was significantly higher in salmon fed 100 % VO compared with salmon fed 65 % VO. In conclusion, the current study suggests that suboptimal dietary levels of cholesterol in combination with low levels of EPA and DHA (1.3 % of FAs) can result in minor metabolic perturbations in the liver of Atlantic salmon.

The use of plant-based feeds has introduced undesirables, such as pesticides, that have previously not been associated with seawater farming of fish species such as Atlantic salmon. Earlier wide-scope chemical screening showed that the organophosphate (OP) pirimiphos-methyl (PM-m) is one of the most prevalent pesticides present in commercially produced Atlantic salmon feed. Information on the safe upper limit of background levels of PM-m in Atlantic salmon feeds with regards to possible adverse effect on fish health is lacking. Therefore, Atlantic salmon (132 ± 25 g) were fed graded levels of PM-m spiked feeds (0, 0.35, 1.5, 5, and 22 mg kg−1) in triplicate for 3 months. Adverse effects were assessed on OP target toxic exposure such as plasma choline esterase (ChE), and secondary toxic responses such as lipid metabolism and oxidative stress, as well as general adverse effect parameters (plasma biochemistry, haematology, and growth). Safe limits were set by model-fitting the effect data in a dose-response (lower bound) bench mark dose (BMDL) regression analysis. Fish fed 1.5 mg kg−1 and higher had a significant (p

Atlantic salmon (Salmo salar) were fed diets with the three vegetable oils, rapeseed oil (RO), palm oil (PO) and soybean oil (SO), in a mixture design for 12 weeks. The diets contained high levels of monounsaturated fatty acids (mainly 18:1n-9), saturated fatty acids (mainly 16:0) and n-6 fatty acids (mainly 18:2n-6), respectively. The dietary content of 20:4n-6 (arachidonic acid, ARA), 20:5n-3 (eicosapentaenoic acid, EPA) and 22:6n-3 (docosahexaenoic acid, DHA) was similar in all diets. The aim was to investigate how dietary 18:1n-9, 16:0 and 18:2n-6 affect the fatty acid composition of salmon tissues, as well as the lipid metabolism and the fish health through lipid-derived compounds such as eicosanoids. Dietary 18:2n-6 had a clear impact on the fatty acid profile of polar lipids, and caused an increase in ARA levels combined with a concomitant decrease in EPA levels, indicating a direct replacement effect. There was a gradient among the tissues, where the dietary effects were most pronounced in liver polar lipids, followed by heart polar lipids and erythrocytes, while retina and brain were the most conserved. The DHA level in polar lipids was highly conserved. Eicosanoid levels in liver were affected by diets, but did not directly reflect the content of precursor fatty acids in the cell membranes, indicating a more complex relationship. Profiling of the main metabolites of the liver revealed distinct differences between fish fed RO and PO compared to fish fed SO, where the latter had reduced levels of both free carnitine and several acylcarnitines. The accumulation of triacylglycerol in the liver also varied between the diet groups, with the level of dietary saturated fat (16:0) appearing to be the variable in the design explaining most of the variation.

Post-smolt Atlantic salmon (Salmo salar) were fed with standard feed added one of five concentrations of either pure deoxynivalenol (DON; 0.5–6 mg/kg) or pure ochratoxin A (OTA; 0.2–2.4 mg/kg), or no added toxins for up to 8 weeks. Performance effects (feed intake, feed efficiency, gain, length and condition factor), various clinical biochemical parameters, packed cell volume and vaccination response against Aeromonas salmonicidae were all inversely correlated with DON dose, whereas relative liver weight increased with DON dose. In fish fed OTA, however, the effects at the doses tested were rather small. We observed no effects of OTA exposure on performance parameters, but some clinical biochemical parameters tended to increase with OTA dose primarily at 3 weeks, and compared with controls OTA exposure caused increased mRNA expression of two immune markers in the spleen. No liver histopathological effects were found from DON or OTA exposure. For DON, we derived a BMDL20 of 0.3 mg/kg feed for reduced total protein in plasma, a BMDL5 of 0.5 mg/kg feed for reduced condition factor, and a NOAEL of 1 mg/kg feed for DON. For OTA, a BMDL or NOAEL could not be derived (>2.4 mg/kg).

The aim of this study was to compare how different dietary vegetable oil n-6/n-3 ratios affect gene responses involved in inflammation, signaling pathways, fatty acid synthesis and oxidation, oxidation and apoptosis as well as eicosanoid production in salmon head kidney tissues and isolated head kidney leukocytes. Salmon smolts (200 g) were fed four different diets where the main lipid components were palm oil (n-6/n-3 ratio = 0.7), rapeseed oil (n-6/n-3 ratio = 0.9), and soybean oil (n-6/n-3 ratio = 2.4) and a high soybean oil diet with an n-6/n-3 ratio = 4. Both head kidney tissue and leukocytes isolated from head kidneys were sampled from the four diets, but from different fish. Leukocytes isolated from the head kidneys were seeded into culture wells and added lipopolysaccharide (LPS) to induce inflammatory responses. Controls without LPS were included. Head kidney leukocytes and the tissues should have the same phenotype reflecting the different diets. Interleukin 1β (IL-1β) transcription was elevated in head kidney tissue and especially in LPS treated leukocytes isolated from soybean oil (n-6/n-3 = 2.4) fed salmon, which confirmed the suitability of the in vitro model in this experiment. Leukocytes, treated with LPS, and isolated from salmon fed the soybean oil diet (n-6/n-3 = 2.4) also upregulated tumor necrosis factor alpha (tnf-α), cyclooxygenase (cox2), prostaglandin D and E synthase (ptgds, ptges), fatty acyl synthase (fas), 5 and 6 desaturases (5des, 6 des) and a fatty acid translocase protein (cd36) when compared to the other diets. The results suggest that diets with a specific n-6/n-3 ratio influence the transcription of pro-inflammatory genes and may be cross-linked to transcription of selected fatty acid metabolism genes. Salmon fed the palm oil diet (n-6/n-3 = 0.7) showed a lower expression of inflammatory genes. Instead, peroxisome proliferator activated receptor β1 (pparβ1), acyl coenzyme A (aco), apoptosis regulator (bax) and superoxide dismutase (sod) were upregulated in leukocytes in vitro, while head kidney tissue transcription of a dendritic marker (cd83) was lower than measured in tissues from fish fed the other diets. The concentration of LTB4 (10-20 ng/mL) were relatively constant in leukocyte supernatants, all diets. Head kidney leukocytes from soybean oil (n-6/n-3 = 2.4) fed fish produced LPS induced PGE2 (mean 0.5 ng/mL) while leukocytes isolated from palm oil diet (n-6/n-3 = 0.7) secreted very high amounts of LTB5 (50-70 ng/mL). In addition, equal amounts of LPS induced PGE2 and PGE3 (mean 0, 5 ng/mL) were produced, indicating that the n-6/n-3 ratio of this saturated fatty acid may have a specific impact on eicosanoid production in the head kidney of salmon.

Post-smolt Atlantic salmon (Salmo salar) were fed standard feed with added 2 or 6 mg kg–1 pure deoxynivalenol (DON), 0.8 or 2.4 mg kg–1 pure ochratoxin A (OTA), or no added toxins for up to 8 weeks. The experiments were performed in duplicate tanks with 25 fish each per diet group, and the feed was given for three 2-h periods per day. After 3, 6 and 8 weeks, 10 fish from each diet group were sampled. In the following hours after the last feeding at 8 weeks, toxin elimination was studied by sampling three fish per diet group at five time points. Analysis of DON and OTA in fish tissues and plasma was conducted by liquid chromatography-mass spectrometry and high-pressure liquid chromatography with fluorescence detection, respectively. DON was distributed to the liver, kidney, plasma, muscle, skin and brain, and the concentrations in liver and muscle increased significantly from 3 to 8 weeks of exposure to the high-DON diet. After the last feeding at 8 weeks, DON concentration in liver reached a maxim...

In two long-term feeding trials in seawater, Atlantic salmon were fed EPA+DHA in graded levels, from 1·3 to 7·4 % of fatty acids (FA, 4–24 g/kg feed) combined with approximately 10 % 18 : 3n-3, at 6 and 12°C. Dietary EPA appeared to be sufficient in all diet groups, as no differences were seen in polar lipid tissue concentrations of either the brain, retina or erythrocytes. For DHA, a reduction in tissue levels was observed with low dietary supply. Effects on brain DHA at ≤1·4 % EPA+DHA of dietary FA and retina DHA at ≤2·7 % EPA+DHA of dietary FA were only observed in fish reared at 6°C, suggesting an effect of temperature, whereas tissue levels of n-6 FA increased as a response to increased dietary n-6 FA in both the brain and the retina at both temperatures. DHA levels in erythrocytes were affected by ≤2·7 % EPA+DHA at both temperatures. Therefore, DHA appears to be the limiting n-3 FA in diets where EPA and DHA are present in the ratios found in fishmeal and fish oil. To assess the physiological significance of FA differences in erythrocytes, the osmotic resistance was tested, but it did not vary between dietary groups. In conclusion, ≤2·7 % EPA+DHA of FA (≤9 g/kg feed) is not sufficient to maintain tissue DHA status in important tissues of Atlantic salmon throughout the seawater production cycle despite the presence of dietary 18 : 3n-3, and effects may be more severe at low water temperatures.

The nutritional requirement for n-3 long-chain PUFA in fast-growing Atlantic salmon (Salmo salar) during grow out in the sea is not well documented. Diets were formulated with levels of EPA (20 : 5n-3) and DHA (22 : 6n-3) ranging from 1·3 to 7·4 % of fatty acids (4–24 g/kg feed). Two long-term trials were conducted through the seawater phase, the first at 6 and 12°C, and the second at 12°C. In the first trial, growth at both temperatures was significantly lower in fish fed 1·4 % EPA+DHA of total fatty acids compared with the 5·2 % EPA+DHA group. In the second trial, growth was significantly lower in fish fed 1·3 and 2·7 % compared with 4·4 and 7·4 % EPA + DHA. Fatty acid composition in the fish reflected diet composition, but only after a 7-fold increase in body weight did the fatty acid profile of the fish stabilise according to dietary fatty acids (shown for EPA and DHA). The retention efficiency of DHA increased with decreasing dietary levels, and was 120–190 and 120–200 % in trials 1 and 2, respectively. The retention efficiency of EPA was lower (60–200 %), and values >100 % were only achieved at the lowest dietary levels in both trials. Temperature did not affect fatty acid retention efficiency. These results suggest that Atlantic salmon have a specific requirement for EPA + DHA >2·7 % of fatty acids for optimal long-term growth in seawater, and that short-term growth trials with less weight increase would not show these effects.

A two-compartmental model for the kinetics of carry-over of the brominated flame retardant α-hexabromocyclododecane (HBCD) from feed to the fillet of farmed harvest-sized Atlantic salmon (Salmo salar L.) was developed. The model is based on a fat compartment for storage of the lipophilic α-HBCD and a central compartment comprising all other tissues. Specific for this model is that the salmon has a continuous growth and that fillet contaminant levels are explained by both the fat and the central compartments. The uptake and elimination kinetics are obtained from experimental data where consumer sized (start weight approximately 1 kg) Atlantic salmon was fed α-HBCD spiked feed (280 ± 11 µg kg−1) for 2 months followed by a depuration period of 3 months. The model was used to simulate the HBCD feed-to-fillet transfer in Atlantic salmon under realistic farming conditions such as the seasonal fluctuations in feed intake, growth and fillet fat deposition. The model predictions gave fillet concentrations of 0.2–1.8 µg kg−1 depending on the level of fish oil inclusion in the salmon diets when using fish oil with high POP background levels. Model simulations show that currently farmed Atlantic salmon can contribute to a maximum of 6% of the estimated provisional food reference dose for HBCD.

Atlantic cod (Gadus morhua), initial weight 15 g, were fed ten experimental diets for 15 weeks. The diets were based on a mixture of plant proteins (PP) and fish meal (FM), where PP constituted 65% of dietary protein. PP mixtures were chosen to reach as low levels of lysine and methionine as possible. The diets were supplemented with increasing amounts of lysine (19.2–31.9 g kg−1 diet) or methionine (9.4–12.3 g kg−1 diet), in a regression design. No growth difference among diet groups was found in the plant-based diets. Increased dietary lysine resulted in decreased liver size, plasma triacylglycerol concentration (TAG) and lipid productive value (LPV). Methionine additions did not result in changed Hepatosomatic index (HSI), LPV or plasma TAG. Feed conversion ratio (FCR) and protein utilization were neither affected by lysine nor methionine. Plasma and muscle concentrations of free lysine and methionine correlated with dietary levels 5-h post feeding. Overall conclusion was that cod maintain growth rates in plant-based diets if dietary protein was high, without additional supplements of crystalline lysine or methionine. Lysine intake significantly influenced lipid metabolism, showing the necessity to add lysine in plant protein-based diets to hinder increased lipid deposition. No such effects were found because of lack of methionine additions.

A regression design with increasing levels of a plant protein mixture replacing fish meal in diets for Atlantic cod, was used to elucidate effects on growth, feed utilization (FCR), digestibility (ADC), and retention of nutrients (PER, PPV and HSI). The plant protein diets contained soybean meal, soy protein concentrate and wheat gluten meal, all of high quality, and were added crystalline methionine and lysine to achieve levels of indispensable amino acids for maximizing growth in other fish species. Initial fish weight was 1652 ± 6 g (mean ± SEM) and the experiment lasted for 28 weeks from December 2004. High growth (SGR 0.31–0.35) and feed utilization (FCR 1.06–1.26) were obtained up to 50% plant protein inclusion. Above this inclusion level, growth (SGR 0.30–0.14) and feed utilization (FCR 1.42–2.71) were reduced. None of the diets affected whole body, liver or muscle proximate compositions. Hepatosomatic index was not effected by diet up to 75% plant protein (HSI 12.7–14.3), but was significantly lower for the 100% plant protein diet group (HSI 10.1–11.4). Plasma and muscle free amino acid pools, sampled 5 h post-feeding, partly reflected diet amino acid composition. Reduced protein retention was found as dietary plant protein increased (PPV 0.45–0.16 and PER 2.11–0.73), however without any reductions in apparent digestibility coefficients. There was a decrease in vitamin B12 concentrations in the diets (120–10 μg kg− 1) as inclusion of plant protein increased, but no specific deficiency symptoms were detected in the fish. Furthermore, the sufficiency of dietary methionine supply in the highest replacement groups, can be questioned based on low levels in the muscle free amino acid pool. Overall conclusion is a great potential for using quite high inclusions of plant proteins in cod diets, provided that the plant ingredients are of high quality.

Atlantic cod were fed 7 different diets from August 2001 to June 2002 varying in protein (range 36–66% of feed), lipid (10–28% of feed) and starch (4–19% of feed) concentration. These dietary treatments were duplicated in netpens with continuous light from metal halogen lamps mounted above surface and under natural light conditions. None of the diets resulted in impaired health of the fish measured as mortality or increased plasma levels of organ specific transaminases or glucose. A few fish in the groups under continuous light conditions spawned during the experimental period while all cod under natural light conditions spawned during spring 2002. Use of continuous light from August 2001 delayed spawning for most of the cod by at least 4 months, this even though the fish was farmed under continuous light from February to August 2001. Spawning decreased final weight in all groups irrespective of diet composition. Postponed sexual maturation will allow continuous growth and thus the cod to reach market size before first spawning. Increased dietary protein increased growth irrespective of light treatment. Liver indices were negatively correlated with increased dietary protein: lipid (P : L) ratio. These results were not affected by spawning. Investment in maturation measured as gonad index in March for the groups kept under natural light conditions was also negatively correlated with increased P : L ratio. Consequently, groups fed diets with the highest dietary protein had the smallest livers and gonads. The present study confirms that dietary protein is the main driver of growth in cod whereas energy deposition is more closely related to dietary lipid content.

To determine the effect of long term feeding of diets with high levels of vegetable oils gilthead seabream juveniles were fed for 7 months with several diets containing vegetable oils at two inclusion levels. Anchovy oil was the only lipid source of FO (fish oil) diet, whereas in diets 60RO, 60LO and 60SO, 60% of fish oil was substituted by rapeseed (RO), linseed (LO) or soybean (SO) oils, respectively, and 80% was substituted by linseed or soybean oils in diets 80LO and 80SO. Afterwards, all fish were fed only a fish oil diet to determine the progressive evolution of fillet fatty acid profiles, since fish muscle fatty acid composition changes by feeding vegetable oils affecting its nutritional value for human health. Results showed that substitution by vegetable oils of up to 60% fish oil in diets for gilthead seabream does not affect growth and feed utilization even after a long feeding period. However, 80% substitution of fish oil significantly reduced growth. Feeding vegetable oils reduced muscle contents of docosahexaenoic acid (DHA) and arachidonic acid (ARA) to a lower degree than their reduction in the diet, denoting their importance, reduction of eicosapentaenoic acid (EPA) in muscle being more pronounced. Moreover, re-feeding with a FO diet for 60 days effectively recovered muscle DHA and ARA contents, whereas those of EPA were not recovered even after 90 days. Linoleic acid was strongly retained even after “wash out”. Fish fillets were very well accepted by the panel of judges. Dietary soybean oil inclusion produced significantly less shiny and less hard fillets, but more juicy and adhesive than FO fillets, with a slight earthy flavour when fish were fed 80SO diet. However linseed oil inclusion only increased shininess when fish were fed 80LO diet. A significantly higher chroma value was obtained for 80LO fish fillets, although it was not noticeable to the human eye.

Juvenile Atlantic cod (Gadus morhua) were fed extruded feeds formulated to contain 360–660 g kg−1 protein, 80–280 g kg−1 lipid and 80–180 g kg−1 starch at feeding frequencies of either once per day or every second day to satiation. The trial was conducted at 8 °C and lasted for 28 weeks during which fish were weighed five times at regular intervals. Sampling for proximate analysis was performed at the start, after 12 weeks and at the end of the trial. Fish grew from an average weight of 192 g to between 750 and 866 g, with growth being negatively affected by low dietary protein concentration. High dietary starch concentrations had some negative effects on growth, whereas changes in dietary fat concentration had no significant effect on growth. Liver indices (at the end of the experiment) varied between 80 and 170 g kg−1, and there was a negative correlation between the ratio of protein to fat and liver index. Feed conversion ratio (FCR) ranged between 0.74 and 0.88, and feed utilization improved with increasing concentrations of dietary protein and fat. Increasing dietary starch concentrations resulted in poorer feed utilization. To achieve good growth and protein retention, and avoid excessive liver size in juvenile cod, feeds should contain 500–600 g kg−1 crude protein, 130–200 g kg−1 lipid and

This study evaluated the effects of fish oil (FO) replacement by vegetable oils [soybean oil (SO), rapeseed oil (RO), linseed oil (LO)] and subsequent feeding with FO on the liver morphology of sea bream. A short-term trial (3 months) and long-term trial (6 months) were carried out feeding sea bream with the following experimental diets: FO100%; SO60% + FO40%; RO60% +FO40%; LO60% + FO40%; SO + RO +LO60% + FO40%. Finally, all groups from the long-term trial were fed with FO100% for 95 days (washout period). Liver samples were taken for histological and biochemical studies. In both the short- and long-term trials, livers of sea bream fed LO60% and SO + RO + LO60% showed a similar hepatic morphology to that observed in fish fed FO100%. In contrast, sea bream fed SO60% showed an intense steatosis, with foci of swollen hepatocytes containing numerous lipid vacuoles. After the washout period, a considerable reduction of the cytoplasmic vacuolation and the lipid vacuole accumulation were observed in the livers of fish fed the different experimental diets. The results of this study suggested that the type of non-essential fatty acid, characteristic of vegetable oils, induces the appearance of steatosis in the following order: linoleic acid > linolenic acid > oleic acid. However, the liver alterations found during the experimental periods with vegetable oils are reversible when the fish are re-fed with a balanced diet (FO100%), indicating the non-pathological character of these histological changes.

As the supply of marine fish oil is becoming a limiting factor in the production of Atlantic salmon (Salmo salar), new diets and alternative sources of energy are being tested. Plant oils are natural potential candidates to replace fish oil, but the different levels of essential polyunsaturated fatty acids may influence the health and growth of salmon. In this study, we have investigated the resistance to transport stress and bacterial infection, phagocytic activity in head kidney macrophages and eicosanoid metabolism in salmon fed three different diets. In high-energy fishmeal based diets, 50% and 100% of the supplementary fish oil (FO) was replaced with soybean oil (SO). The three dietary groups were fed for 950 day-degrees at 5 � C (27 weeks) and 12 � C (11 weeks) before challenging the fish with Aeromonas salmonicida, analyzing the lipid composition of head kidney and examining macrophage function in vivo and in vitro. Dietary fatty acids affected the lipid composition of the kidney. The level of eicosanoid precursor’s 20:4n-6 and 20:3n-6 were 3 and 7-fold higher in the 100% SO group compared with the FO group. The total fraction of n-3 lipids in kidney was 19% in the SO group, compared to 16% and 12% in the 50% or 100% SO groups, respectively. However, the production of leucotriene B4 (LTB) and prostaglandin E2 (PGE) immunoreactive materiel from exogenously added arachidonic acid in head kidney macrophages was only affected by the composite diet (increased) at 5 � C. In addition, the phagocytic activity of kidney macrophages in vivo and in vitro was not affected by diet. No effect of diet was observed on transport stress or susceptibility to a bacterial infection with Aeromonas salmonicida. Atlantic salmon therefore seems to tolerate a diet solely based on soybean oil as lipid source, without any detrimental effects on growth, health and immune functions.

Effect of season and diet on muscle composition were evaluated in farmed Atlantic cod (Gadus morhua L.), fed varying levels of macro-nutrients, and kept at two different light regimes during 1 year grow-out in sea-cages. The cod were fed seven different diets varying in protein, lipid and starch, in a mixture design. The diets spanned 4–20% starch, 8–26% lipid and 36–66% protein. Each dietary regime was subjected to two different light regimes: continuous light (24 h), or natural light (August 2001 to June 2002). Fish subjected to natural light started to mature in December/January and spawning was more or less completed during March/April. No maturation was registered in the continuous light groups at this point. No variation was found in muscle dry matter, protein or lipid concentration as a consequence of the dietary or light regime variations, except for the groups spawning in March. Glycogen varied from 1 to 6.5 mg g−1 wet weight, without any correlation to the present dietary variations. At the June 2002 sampling all groups given a natural light regime showed almost twice the concentration of muscle glycogen compared with fish subjected to continuous light. Such clear results were not measured at the December or March samplings. Increased dietary lipid resulted in lowered muscle vitamin E concentration. A strong covariation between muscle vitamins C and E was found at all samplings, and these showed a negative correlation towards eicosapentaenoic (EPA) and docosahexaenoic acid (DHA), and the ratio n-3/n-6 in muscle. The highest dietary vitamin E resulted in the lowest muscle vitamin C concentrations. All dietary lipid added was identical and of marine origin; in addition, the wheat added as a starch source summed up parts of the dietary lipid fraction, resulting in slightly decreased sum of polyenes in the diets holding the highest levels of wheat. The lean cod muscle consists almost solely of membrane lipids. These were, however, highly influenced by the dietary lipid composition, especially as concerned the different monoenes. Two of the diets showed tendencies to increased thiobarbituric acid-reactive substances (TBARS). This was not reflected in the muscle concentrations of vitamins E or C, and did not result in any changes in muscle TBARS values at any of the samplings, except for the fish spawning in March. The water-soluble fraction of the muscle protein, pH range from 4.5 to 6, did show a difference in structure when comparing our experimental cod to wild cod. We could also identify a differential pattern between some of the experimental groups. The method used to identify this was, however, not quantitative, and further studies are needed. Taste panel evaluation and shear force measurements after final sampling in June concluded with minor differences between muscles from the different diet groups, except for fish given one diet with relatively high protein, intermediate lipid and low starch levels. Fish kept at continuous light was described as less firm (texture) compared with fish kept at natural light, explained partly by the different size of these two categories of fish. In conclusion, both diet and season (spawning or not) did influence several of the measured muscle parameters.

Background To ensure sustainable aquaculture, fish derived raw materials are replaced by vegetable ingredients. Fatty acid composition and contaminant status of farmed Atlantic salmon (Salmo salar L.) are affected by the use of plant ingredients and a spillover effect on consumers is thus expected. Here we aimed to compare the effects of intake of Atlantic salmon fed fish oil (FO) with intake of Atlantic salmon fed a high proportion of vegetable oils (VOs) on development of insulin resistance and obesity in mice. Methodology/principal findings Atlantic salmon were fed diets where FO was partly (80%) replaced with three different VOs; rapeseed oil (RO), olive oil (OO) or soy bean oil (SO). Fillets from Atlantic salmon were subsequently used to prepare Western diets (WD) for a mouse feeding trial. Partial replacement of FO with VOs reduced the levels of polychlorinated biphenyls (PCB) and dichloro-diphenyl-tricloroethanes (DDT) with more than 50% in salmon fillets, in WDs containing the fillets, and in white adipose tissue from mice consuming the WDs. Replacement with VOs, SO in particular, lowered the n−3 polyunsaturated fatty acid (PUFA) content and increased n−6 PUFA levels in the salmon fillets, in the prepared WDs, and in red blood cells collected from mice consuming the WDs. Replacing FO with VO did not influence obesity development in the mice, but replacement of FO with RO improved glucose tolerance. Compared with WD-FO fed mice, feeding mice WD-SO containing lower PCB and DDT levels but high levels of linoleic acid (LA), exaggerated insulin resistance and increased accumulation of fat in the liver. Conclusion/Significance Replacement of FO with VOs in aqua feed for farmed salmon had markedly different spillover effects on metabolism in mice. Our results suggest that the content of LA in VOs may be a matter of concern that warrants further investigation.

Diets with increasing levels of potato and maize starch were fed to duplicate groups of maturing cod, Gadus morhuaL., from June 1991 to January 1992.The level of starch in the diet did not influence feed conversion factors, broodfish growth or gonadal development. Low levels of fillet glycogen were measured at all samplings, without variations according to dietary starch, or to developmental stage of the brood fish. During the reproductive phase no decreases were measured in relative liver size or liver protein, lipid or glycogen, indicating no net use of stored energy from the liver to build up gonads as long as the fish was offered feed. The variations in dietary starch vs. protein did not alter the composition of gonad dry matter, protein, lipid or glycogen levels. However, dry matter and protein levels increased during maturation in all groups. Plasma glucose levels were within normal ranges in all dietary groups at all sampling times, indicating no negative effects of high levels of dietary starch in brood fish, as also confirmed by stable and normal haematocrit, red blood cell count, and mean cell volume in blood. Haemoglobin, mean cell haematocrit and mean cell haemoglobin concentration decreased during the reproductive phase. Serum haemolytic complement activity did not vary between dietary treatments. Glycogen, dry matter, protein and lipid levels in eggs did not vary between dietary treatments of brood fish. Very low levels of glycogen were measured, suggesting that this energy reserve was of minor importance. Dry matter and lipid levels were quite stable in relation to egg developmental stage: protein levels decreased from day 0 until hatching.

The effect of soybean oil (SO) and fish oil (FO) on the relative molecular species distribution of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylserine (PS) in Atlantic salmon head kidney was studied using normal-phase liquid chromatography coupled with negative ion electrospray tandem mass spectrometry. The conformation of identity of the phospholipid species was based on retention time, the mass of the [M−H] − ([M−15] − for PC) molecular ions and the carboxylate anion fragments in the product ion spectrum. The intensity ratio of sn -1/ sn- 2 fragment ions increased with increasing number of double bonds in the sn- 2 acyl chain but was not affected by increasing number of double bonds in the sn- 1 acyl chain of the species examined. The relative distribution of the molecular species was determined by multiple reaction monitoring of the carboxylate anion fragment from the sn -1 position. A total of 68 different phospholipid species were determined in the head kidney and the largest amount was found in PE (22 species). Depending on the diet, the main species identified in the different phospholipid classes were; PC 16:0/18:1, PE 16:0/22:6, PI 18:0/20:4 and PS 16:0/22:6. The SO diet significantly increased the 18:2, 20:3 and most 20:4 containing species and significantly reduced the 14:0 and most 20:5 and 22:6 fatty acid containing species. The increase of the 20:4 and the decrease of the 20:5 and 22:6 containing species were dependent on the fatty acid combination of the species

Assimilation and elimination rate constant of dietary polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DLPCBs) with a World Health Organization toxic equivalency factor (WHO-TEF) were estimated in market-size Atlantic salmon (Salmo salar) using fish that were previously fed vegetable oil-based (low in PCDD/Fs and DLPCBs) or fish oil-based (high in PCDD/Fs and PCBs) diets. At the start of the kinetic trial, half the fish that were fed fish oils were fed vegetable oil feeds and inverted (cross-over design) for five months. The assimilation efficiencies of the PCDD/F congeners were more variable (3-89%) and, generally, were lower than those of the DLPCBs (70-80%). Among the PCDD/F congeners, the assimilation efficiency of the most toxic tetra- and pentachlorinated PCDD/Fs was greater than that of higher-chlorinated PCDD/Fs. Elimination rates for DLPCBs were higher than those for PCDD/Fs. Lower-chlorinated PCDDs had a lower elimination rate than the higher-chlorinated PCDDs, but no differences were observed among PCDF congeners or DLPCB congeners. Kinetic parameters were used to predict the level of WHO-TEF dioxins and DLPCBs in Atlantic salmon reared in a large-scale facility under commercial conditions. Predictions were based on preanalyzed levels of these organochlorines in feeds with three different replacement levels (0, 30, and 60%) of vegetable oil. A simple one-compartmental, first-order kinetic model was used to predict the level of sum WHO toxic equivalents for PCDD/Fs and DL-PCBs. The predicted values varied by 0 to 11% from the measured values in the commercially reared salmon.

1. 1. Cod ( Gadus morhua ) were fed diets containing protein energy to total energy levels (PE/TE) of 10.0, 20.6, 29.6, 38.4, 56.2 and 74.1% for 21 days. 2. 2. Ribosomes were isolated from the white trunk muscle tissue, the capacity for protein synthesis in vitro determined and related to muscle tissue wet weight, rRNA and DNA. 3. 3. Protein concentrations of less than 47.4% PE/TE in the diets reduce the ribosomal capacity for protein synthesis per g wet weight and per mg DNA, and the tissue contents of rRNA and ratio of rRNA/DNA. 4. 4. The capacity for muscle protein synthesis in vitro is a significant and sensitive parameter of protein inadequacy in fish diets.

The present study was designed to investigate potential effects of arachidonic acid (ARA) on the reproductive physiology of female Atlantic cod (Gadus morhua L.). Two-year old Atlantic cod of both sexes were equally distributed into eight sea cages after completion of their first spawning in May 2005. Four experimental groups were established and fed diets with different levels of ARA corresponding to 0.5, 1, 2 and 4% of total fatty acid. Ovarian growth and development was documented every month. Fatty acid composition was analysed in ovaries, liver and plasma at the beginning of the experiment, one month prior to spawning, and in spent fish, one month after spawning was completed. Plasma concentrations of estradiol-17β, testosterone and vitellogenin, and ovarian gene transcript levels of steroidogenic acute regulatory protein (star), P450aromatase (cyp19a1a) and 20β-hydroxy steroid dehydrogenase (20bhsd/cbr1) were monitored every month in fish fed the experimental diets and related to oocyte stage. Potential fecundity was calculated based on ovarian samples taken one month before onset of spawning. Ovarian and plasma ARA levels were highly correlated to dietary ARA levels. There was a net accumulation of ARA compared to other essential fatty acids in ovarian tissue that was reflected in a decrease in EPA:ARA ratio. Plasma concentrations of vitellogenin, estradiol-17β and testosterone and key gene transcript levels were affected by dietary ARA and stage of maturation. The results show that ARA has a significant influence on the reproductive physiology of female Atlantic cod.

Few ingredients allow the complete replacement of fish meals (FM) and fish oils (FO) in aquaculture feeds without affecting fish performance or fillet nutritive value. This is due to the adequate content of essential nutrients, including the n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), and the unique high palatability of FM and fish oil. Some microalgae present abundant amounts of these fatty acids, for instance docosahexaenoic acid (DHA). Therefore, the aim of the present study was to evaluate the effect of two different microalgae products, one providing DHA and eicosapentaenoic acid (EPA, 20:5n-3; ED diet), and the other one DHA and n-6 docosapentaenoic acid (DPA, 22:5n-6; DD diet), in combination with poultry oil and rapeseed oil, as total replacers of fish oil, at two different dietary fish meal contents (15 and 7.5%). The effects of these dietary oil combinations on performance, composition and nutritive quality indexes of gilthead sea bream (Sparus aurata) juveniles were studied and compared against a positive control diet (FO) and two negative control diets (PO diets), one for each dietary fish meal content tested, giving in total 7 experimental diets. Both microalgae products in combination with poultry and rapseed oils were able to completely replace fish oil in practical diets with 15% FM without affecting growth performance, utilization of dietary fatty acids or the nutritional quality of fish fillet for the consumer. On the contrary, PO alone was not able to completely replace fish oil and negatively affected fish performance, in relation to an insufficient dietary n-3 LC-PUFA content. A similar decrease in growth performance was also observed with the reduction of the dietary FM content to 7.5%. In conclusion, both oils from microalgae, providing either DHA and EPA or DHA and n-6 DPA, were effective n-3 LC-PUFA sources for sea bream juveniles and allowed complete replacement of fish oil in combination with more cost-effective lipid sources, such as poultry and rapeseed oils.