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The present study evaluated the effects of increasing the dietary levels of EPA and DHA in Atlantic salmon (Salmo salar) reared in sea cages, in terms of growth performance, welfare, robustness and overall quality. Fish with an average starting weight of 275 g were fed one of four different diets containing 10, 13, 16 and 35 g/kg of EPA and DHA (designated as 1·0, 1·3, 1·6 and 3·5 % EPA and DHA) until they reached approximately 5 kg. The 3·5 % EPA and DHA diet showed a significantly beneficial effect on growth performance and fillet quality compared with all other diets, particularly the 1 % EPA and DHA diet. Fish fed the diet containing 3·5 % EPA and DHA showed 400–600 g higher final weights, improved internal organ health scores and external welfare indicators, better fillet quality in terms of higher visual colour score and lower occurrence of dark spots and higher EPA and DHA content in tissues at the end of the feeding trial. Moreover, fish fed the 3·5 % EPA and DHA diet showed lower mortality during a naturally occurring cardiomyopathy syndrome outbreak, although this did not reach statistical significance. Altogether, our findings emphasise the importance of dietary EPA and DHA to maintain good growth, robustness, welfare and fillet quality of Atlantic salmon reared in sea cages.

The present study evaluated the effects of increasing the dietary levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in Atlantic salmon (Salmo salar) reared in sea cages, in terms of growth performance, welfare, robustness, and overall quality. Fish with an average starting weight of 275 g were fed one of four different diets containing 10, 13, 16, and 35 g/kg of EPA and DHA (designated as 1.0%, 1.3%, 1.6%, and 3.5% EPA and DHA) until they reached approximately 5 kg. The 3.5% EPA and DHA diet showed a significantly beneficial effect on growth performance and fillet quality compared with all other diets, particularly the 1% EPA and DHA diet. Fish fed the diet containing 3.5% EPA and DHA showed 400-600 g higher final weights, improved internal organ health scores and external welfare indicators, better fillet quality in terms of higher visual colour score and lower occurrence of dark spots, and higher EPA and DHA content in tissues at the end of the feeding trial. Moreover, fish fed the 3.5% EPA and DHA diet showed lower mortality during a naturally occurring cardiomyopathy syndrome outbreak, although this did not reach statistical significance. Altogether, our findings emphasise the importance of dietary EPA and DHA to maintain good growth, robustness, welfare, and fillet quality of Atlantic salmon reared in sea cages.

There is an increased use of vegetable oils containing n-6 fatty acids (FA) in aquafeeds, and several trials indicate that there might be an increased requirement of EPA and DHA for Atlantic salmon when they are fed higher dietary n-6 FA. With a limited supply of EPA and DHA for production of aquafeeds, it is important to know how to efficiently use these FA to maintain growth and health of the fish. In the present trial, three diets containing equal amounts of n-3 FA (about 7·7 % of total FA) and different n-6:n-3 FA ratios (about 1, 2 and 6), as well as one diet with n-6:n-3 FA ratio at about 1 but twice as much n-3 FA, were fed to Atlantic salmon. Despite constant dietary n-3, increasing dietary n-6 led to significantly reduced n-3 in tissue polar lipids. Interestingly, EPA was significantly reduced while DHA was not. Maintaining a stable n-3 content in the polar lipids when increasing dietary n-6 FA was only obtained by simultaneously increasing the dietary n-3 content and with this maintaining the same n-6:n-3 FA ratio. Polar lipid n-6 FA in tissues thus primarily reflected the dietary n-6:n-3 FA ratio and not the absolute dietary n-6 FA content. Neutral lipids, on the other hand, reflected the dietary absolute levels of both n-3 and n-6 FA. This study indicates that a better use of dietary EPA is achieved by keeping the dietary n-6:n-3 FA ratio low.

Atlantic salmon were fed diets containing graded levels of EPA + DHA (1·0, 1·3, 1·6 and 3·5 % in the diet) and one diet with 1·3 % of EPA + DHA with reduced total fat content. Fish were reared in sea cages from about 275 g until harvest size (about 5 kg) and were subjected to delousing procedure (about 2·5 kg), with sampling pre-, 1 h and 24 h post-stress. Delousing stress affected plasma cortisol and hepatic mRNA expression of genes involved in oxidative stress and immune response, but with no dietary effects. Increasing EPA + DHA levels in the diet increased the trace mineral levels in plasma and liver during mechanical delousing stress period and whole body at harvest size. The liver Se, Zn, Fe, Cu, and Mn and plasma Se levels were increased in fish fed a diet high in EPA + DHA (3·5 %) upon delousing stress. Furthermore, increased dietary EPA + DHA caused a significant increase in mRNA expression of hepcidin antimicrobial peptide (HAMP), which is concurrent with downregulated transferrin receptor (TFR) expression levels. High dietary EPA + DHA also significantly increased the whole-body Zn, Se, and Mn levels at harvest size fish. Additionally, the plasma and whole-body Zn status increased, respectively, during stress and at harvest size in fish fed reduced-fat diet with less EPA + DHA. As the dietary upper limits of Zn and Se are legally added to the feeds and play important roles in maintaining fish health, knowledge on how the dietary fatty acid composition and lipid level affect body stores of these minerals is crucial for the aquaculture industry.

Dietary electrolyte balance is the equilibrium of monovalent cations and anions that influence the acid-base balance of the feed (dEB = Na + K − Cl, mEq kg−1). Dietary electrolytes/minerals can influence the physiological changes during smoltification in Atlantic salmon. In this context, we aimed to study if the dEB of the freshwater feeds can be used to pre-adapt the hypoosmotic functionality and the associated effects on mineral metabolism. The dEB of commercial freshwater Atlantic salmon feeds in Norway varied from −9 to 400 mEq kg−1 feed. Three experimental feeds were formulated to study incremental levels of dEB reflecting the low (L-dEB, −50 to 0), median (M-dEB, 200–250) and high (H-dEB, 350–400). Triplicate groups of Atlantic salmon parr (36 g) were fed one of the three feeds for 8 weeks in freshwater at 12 °C. The fish were transferred to full strength seawater in indoor tanks and fed a commercial diet for 6 weeks. Growth was not differentially affected by dEB levels, neither in the freshwater phase nor in the seawater. Plasma electrolytes (Na+ and Cl−) and gill mRNA expression of sodium potassium ATPase (NKA a1b, seawater isoform) were significantly lower in L-dEB fed fish. In the intestine, carbonate precipitates 24 h after seawater transfer was higher in fish fed both L-dEB and H-dEB feeds compared to the M-dEB fed fish. Whole body and plasma mineral levels were significantly affected by dEB levels in freshwater feeds. Interestingly, the carryover effect of dEB in freshwater feeds was significant after 6 weeks in seawater for plasma and whole-body Zn status, with the H-dEB fed fish showing significantly increased body Zn status compared to L-dEB and M-dEB fed fish. The study revealed that mineral metabolism and intestinal response to seawater transfer can be pre-adapted by modulating the electrolyte and/or mineral balance in freshwater feeds in Atlantic salmon. Further, dEB did not affect long term development of cataract or vertebral deformities.

The aim of the study was to investigate the influence of dietary level and ratio of n-6/n-3 fatty acids (FA) on growth, disease progression and expression of immune and inflammatory markers in Atlantic salmon (Salmo salar) following challenge with Paramoeba perurans. Fish (80 g) were fed four different diets with different ratios of n-6/n-3 FA; at 1.3, 2.4 and 6.0 and one diet with ratio of 1.3 combined with a higher level of n-3 FA and n-6 FA. The diet with the n-6/n-3 FA ratio of 6.0 was included to ensure potential n-6 FA effects were revealed, while the three other diets were more commercially relevant n-6/n-3 FA ratios and levels. After a pre-feeding period of 3 months, fish from each diet regime were challenged with a standardized laboratory challenge using a clonal culture of P. perurans at the concentration of 1,000 cells L−1. The subsequent development of the disease was monitored (by gross gill score), and sampling conducted before challenge and at weekly sampling points for 5 weeks post-challenge. Challenge with P. perurans did not have a significant impact on the growth of the fish during the challenge period, but fish given the feed with the highest n-6/n-3 FA ratio had reduced growth compared to the other groups. Total gill score for all surfaces showed a significant increase with time, reaching a maximum at 21 days post-challenge and declined thereafter, irrespective of diet groups. Challenge with P. perurans influenced the mRNA expression of examined genes involved in immune and inflammatory response (TNF-α, iNOS, IL4-13b, GATA-3, IL-1β, p53, COX2 and PGE2-EP4), but diet did not influence the gene expression. In conclusion, an increase in dietary n-6/n-3 FA ratio influenced the growth of Atlantic salmon challenged with P. perurans; however, it did not alter the mRNA expression of immune genes or progression of the disease.

Limited availability of fish oils (FO), rich in n-3 long-chain (≥C20) PUFA, is a major constraint for further growth of the aquaculture industry. Long-chain n-3 rich oils from crops GM with algal genes are promising new sources for the industry. This project studied the use of a newly developed n-3 canola oil (DHA-CA) in diets of Atlantic salmon fingerlings in freshwater. The DHA-CA oil has high proportions of the n-3 fatty acids (FA) 18 : 3n-3 and DHA and lower proportions of n-6 FA than conventional plant oils. Levels of phytosterols, vitamin E and minerals in the DHA-CA were within the natural variation of commercial canola oils. Pesticides, mycotoxins, polyaromatic hydrocarbons and heavy metals were below lowest qualifiable concentration. Two feeding trials were conducted to evaluate effects of two dietary levels of DHA-CA compared with two dietary levels of FO at two water temperatures. Fish increased their weight approximately 20-fold at 16°C and 12-fold at 12°C during the experimental periods, with equal growth in salmon fed the FO diets compared with DHA-CA diets. Salmon fed DHA-CA diets had approximately the same EPA+DHA content in whole body as salmon fed FO diets. Gene expression, lipid composition and oxidative stress-related enzyme activities showed only minor differences between the dietary groups, and the effects were mostly a result of dietary oil level, rather than the oil source. The results demonstrated that DHA-CA is a safe and effective replacement for FO in diets of Atlantic salmon during the sensitive fingerling life-stage.

The aim of the current study was to investigate how freshwater diets impact on immunity in Atlantic salmon smolts in freshwater, during transfer to seawater and in post smolts during the seawater stage with and without pancreas disease (PD) infection. Three specific freshwater diets were prepared: (i) A diet similar in composition to commercial salmon freshwater diets (Standard diet); (ii) A diet composed of vegetable oils (rapeseed, palm and linseed oils) mimicking the fat composition in aquatic insects - the natural diet of wild salmon in freshwater (Fatty acid diet); (iii) A diet enriched with possible immune modulating amino acids including dl-methionine, l-lysine, l-threonine and taurine (Amino acid diet). After seawater transfer, all fish were fed the same commercial diet. The head kidneys were extracted, and their leukocytes were isolated from smolts right before transfer to seawater, from post smolts one and six weeks after transfer to seawater, and from post smolts in seawater after 8 weeks of ongoing PD infection. In addition, to provoke bacterial or virus induced inflammation in vitro, the individual leukocyte suspension from all fish were stimulated by lipopolysaccharide (LPS) or polyinosinic acid: polycytidylic acid (PIC). The transfer of smolts from fresh-to seawater changed the transcription of several types of genes. Particularly in isolates from fish fed the Standard or Fatty acid diet in freshwater, overall gene transcription (IL-1β, CD83, INF-γ, cox2, cd36, MGAT2, catalase) declined. However, the Amino acid diet stimulated the LPS induced gene transcription of IL-1β, CD83, Cox2, and INF-γ at this stage. In freshwater smolts, PIC stimulated leukocytes showed higher transcription level of Mx and viperin in the Fatty acid and Amino acid diet groups compared to the Standard diet group. In seawater post smolts, Mx and viperin responded similarly to PIC challenge in all diet groups. Furthermore, leukocytes isolated from PD infected fish, continued responding to PIC, regardless of freshwater diet.

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 loss of fish in the seawater (SW) phase of Atlantic salmon farming is high, and a major proportion of this loss occurs in the period just after SW transfer. In the current study, we hypothesize that improvements made to the diet during the freshwater (FW) stage affect fish growth, survival and robustness later in the SW stage. To test this, salmon parr were fed five experimental diets in FW at 12 °C. In addition to a commercial-like control diet, fish were fed a diet with changed FA composition aimed to be more like the natural feed of salmon in FW, a diet with increased concentrations of selected AA/N-compounds (methionine, lysine, threonine and taurine), a diet with increased concentrations of methionine and certain B-vitamins (folate, B12 and B6) and a final diet combining all of these potential improvements. At the time of SW transfer, the robustness of fish fed the different diets was tested by direct transfer to SW at three different temperatures (8, 12 and 16 °C, without prior acclimation), as well as transfer into open net pens, while fed on a common commercial diet. Growth and proximate composition of the fish did not differ between the diet groups. All diet groups seemed to handle transfer to SW well, and while SW transfer elicited a stress response in the fish, this was not significantly different between diet groups. Fish transferred to SW at 8 °C had higher mortality, reduced mucus layer and increased prevalence of scale loss and wounds, but this applied to all diet groups. Hence, direct transfer to SW at a lower temperature than the fish has been acclimated to cannot be recommended. At the two highest temperatures, there were some differences between the groups in the severity of cataracts. Apart from this, none of the health- or welfare related parameters measured showed any difference between the diet groups, indicating that the control diet was already sufficient.

The salmon louse, Lepeophtheirus salmonis, is a parasitic copepod infecting wild and farmed salmonid fishes in the northern hemisphere. It has a direct lifecycle with a planktonic dispersal phase and an infective copepodid stage preceding five host bound stages. Several models predicting the dispersal of this ecologically and economically important pathogen have been developed, but none include variability in capability to infect. Therefore, the effect of age and temperature on infectivity and lipid metabolism was investigated experimentally using seven synchronized cohorts of copepodids at 5, 10 and 15 °C. In newly molted copepodids infectivity initially increase and then decrease with senescence. Within the experimental temperature range, peak infectivity was higher and occurred earlier at higher temperatures. While degree-days may serve as a useful crude descriptor of developmental age, it did not allow accurate prediction of infectivity peak timing and magnitude unless temperature was included as a separate factor in the derived infectivity model. Senescence was reflected in lipid store depletion and a temperature dependent variability in membrane lipid composition was evident. Interestingly, copepodids developing at 5 °C had approximately 50% less storage lipids when they molted into the parasitic stage than those developing at 10 and 15 °C. The declines in infectivity and storage energy were mirrored in decreasing copepodid swimming activity. When incorporating the copepodid infectivity results from this study into salmon louse dispersal model parameterization, the predictions suggest that earlier models may have underestimated the seasonal differences in salmon louse infection risk. publishedVersion

Several feeding trials with Atlantic salmon fed naturally high phytosterol concentrations due to dietary rapeseed oil inclusion have shown changes in lipid metabolism and increased hepatic lipid storage in the fish. An in vitro trial with Atlantic salmon hepatocytes was, therefore, performed to study the possible direct effects of phytosterols on lipid storage and metabolism. The isolated hepatocytes were exposed to seven different sterol treatments and gene expression, as well as lipid accumulation by Oil Red O dyeing, was assessed. Fucosterol, a sterol found in many algae species, had an effect on the size of individual lipid droplets, leading to smaller lipid droplets than in the control without added sterols. A sterol extract from soybean/rapeseed led to an increase in the percentage of hepatocytes with visible lipid droplets at 20× magnification, while hepatocytes of both the sterol extract-treated groups and fucosterol-treated groups had a larger proportion of their area covered with lipids compared to control cells. Brassicasterol, a sterol characteristic of rapeseed oil, was the only sterol treatment leading to a change in gene expression, affecting the expression of the nuclear receptors, peroxisome proliferator-activated receptor gamma (pparg) and retinoid X receptor (rxr). The current study thus shows that phytosterols can have direct, although subtle, effects on both hepatic lipid storage and gene expression of Atlantic salmon in vitro.

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.

Atlantic salmon (Salmo salar) feeds have changed drastically in their composition from being predominantly marine-based to plant-based. This has altered the dietary supply and availability of micro-nutrients to Atlantic salmon. The impact of graded inclusion levels of a nutrient package (NP) comprising of 25 different micro-nutrients were studied in Atlantic salmon parr in freshwater (Trial 1) and post-smolts in seawater (Trial 2). In brief, the NP was included from 0 to 400%, where 100% corresponded to the recommendation by the National Research Council, 2011. Micro-nutrients, namely Zn, Mn, Se, Cu, Fe, Co, I and vitamin D3 were included in the NP with the objective of (re)evaluating the dietary need to meet the requirement of Atlantic salmon parr and post-smolt, when fed low fish meal, plant ingredient-based diets. Responses in apparent availability coefficient (AAC), whole body and vertebrae mineral concentrations, and retention were analysed. AAC of Cu, Mn, Se and Zn responded in a quadratic fashion with an increase in NP from 0 to 400% in freshwater parr; AAC could not be measured in post-smolt salmon. The whole-body concentration of Zn, Se, Co and I in Atlantic salmon parr were significantly affected by increasing NP inclusion; the same was observed for Zn, Se and Co in post-smolt Atlantic salmon. Vertebrae mineral concentration as the response criterion was non-responsive in parr; whereas, in post-smolt, Co had a linear increase, while Zn and Se showed a non-linear increase upon 0 to 400 NP inclusion. Zinc concentration and activities of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) in vertebrae indicated increased bone resorption in post-smolt Atlantic salmon; TRAP activity increased linearly with NP inclusion in post-smolt, but not in parr. Significant correlations between Zn and Se were observed in AAC and vertebral concentrations, indicating an interaction in intestinal uptake and vertebral deposition. Overall, Atlantic salmon parr held in freshwater were able to satisfy the requirement for the trace minerals Zn, Mn, Se, Cu, and Fe through supply from 100–150 NP, corresponding to 101–132, 47–63, 0.6–0.8, 12–16 and 150–166 mg kg −1, respectively; for iodine, dietary supply from 150–200 NP, corresponding to 0.7–1.6 mg kg−1, was required. In the seawater, Atlantic salmon post-smolt, in general, required micro-minerals and vitamin D3 levels as supplied through 150–200 NP, corresponding to 140–177, Zn; 61–67, Mn; 0.9–1, Se; 14–16, Cu; and vitamin D3, 0.06–0.09 mg kg −1 to fulfil the requirement, except for Cu which was satisfied at 100–150 NP, equivalent to 13–14 mg kg −1 diet.

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

As fish oil is an increasingly limited input-factor, content of the n-3 fatty acids eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) is being gradually reduced in feeds for farmed salmon, and consequences for fish health and fillet quality need to be addressed. In the present study, Atlantic salmon (Salmo salar L.) were fed two levels of EPA and DHA through the seawater rearing period from six weeks after smolt transfer (~ 150 g) until harvest (~ 5000 g). The trial was conducted under commercial production conditions, with triplicate net pens used for each diet group. The control (standard) diet contained 8% EPA + DHA of total fatty acids (FAs, 26 g kg− 1 feed), while the low n-3 diet contained 6% EPA + DHA of FAs until 1200 g body weight and 4.5% in following feeds, averaging out to 5% of FAs (16 g kg− 1 feed) during the entire production cycle. Growth performance and measured health parameters including cataract, liver lipid accumulation and vaccine side-effects did not differ significantly between the two diet groups. Through the production period, the fish faced stressful and challenging conditions such as five treatments against sea lice (one oral and four bath treatments), as well as an outbreak of pancreas disease and considerable mortality from gill infections. Despite this, mortality did not differ significantly between the diet groups, indicating that fish fed low dietary n-3 FAs were equally robust as fish fed the standard diet. Quality classification at slaughter did not reveal any diet effects, and neither did sensory evaluation of the fish fillets. As expected, the fillet FA profile differed, with 5.4% EPA + DHA of total FA (0.9 g 100 g− 1) in the fillet of fish fed the low n-3 diet and 7.5% (1.3 g 100 g− 1) in fish fed the standard diet. The fish fed low EPA + DHA showed a higher frequency of fillet melanin spots, 28.2%, compared to 21.5% in the salmon fed the standard diet, possibly related to dietary FA composition affecting inflammatory processes. Statement of relevance Supply of fish oil is limiting growth of the salmon industry

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

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

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.

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.

The European Food Safety Authority (EFSA) published a risk assessment of erucic acid (22:1n-9) in 2016, establishing a Tolerable Daily Intake (TDI) for humans of 7 mg kg&minus, 1 body weight per day. This report largely excluded the contribution of erucic acid from fish and seafood, due to this fatty acid often not being reported separately in seafood. The Institute of Marine Research (IMR) in Norway analyzes erucic acid and has accumulated extensive data from analyses of fish feeds, farmed and wild fish, and seafood products. Our data show that rapeseed oil (low erucic acid varieties) and fish oil are the main sources of erucic acid in feed for farmed fish. Erucic acid content increases with total fat content, both in farmed and wild fish, and it is particularly high in fish liver, fish oil, and oily fish, such as mackerel. We show that the current TDI could be exceeded with a 200 g meal of mackerel, as at the maximum concentration analyzed, such a meal would contribute 143% to the TDI of a 60 kg person. These data cover a current knowledge gap in the scientific literature regarding the content of erucic acid in fish and seafood.

There was an error published in J. Exp. Biol. (2018) 221 , [jeb161521][1] (doi: [10.1242/jeb.161521][2]). The y -axis labels in [Fig. 3][3] should have the units ‘g per 100 g fillet’. The corrected figure appears below. ![Fig. 3.][4] Fig. 3. The author apologises to the readers for

‘Are we what we eat?’ Yes and no. Although dietary fat affects body fat, there are many modifying mechanisms. In Atlantic salmon, there is a high level of retention of the n-3 fatty acid (FA) docosahexaenoic acid (DHA, 22:6n-3) relative to the dietary content, whereas saturated FAs never seem to increase above a specified level, which is probably an adaptation to low and fluctuating body temperature. Net production of eicosapentaenoic acid (EPA, 20:5n-3) and especially DHA occurs in salmon when dietary levels are low; however, this synthesis is not sufficient to maintain EPA and DHA at similar tissue levels to those of a traditional fish oil-fed farmed salmon. The commercial diets of farmed salmon have changed over the past 15 years towards a more plant-based diet owing to the limited availability of the marine ingredients fish meal and fish oil, resulting in decreased EPA and DHA and increased n-6 FAs. Salmon is part of the human diet, leading to the question ‘Are we what the salmon eats?’ Dietary intervention studies using salmon have shown positive effects on FA profiles and health biomarkers in humans; however, most of these studies used salmon that were fed high levels of marine ingredients. Only a few human intervention studies and mouse trials have explored the effects of the changing feed composition of farmed salmon. In conclusion, when evaluating feed ingredients for farmed fish, effects throughout the food chain on fish health, fillet composition and human health need to be considered.

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.

In the present study, zebrafish (Danio rerio) were fed casein/gelatin-based diets containing either 19 % Bt (Bacillus thuringiensis)-maize or its parental non-Bt (nBt)-maize control for two generations (F0: sixty fish; F1: forty-two to seventy fish per treatment). The study focused on growth and reproductive performance, liver CuZn superoxide dismutase (SOD) enzyme activity, gene transcript levels targeting important cellular pathways in the liver and mid-intestine, histomorphological evaluation of the intestine, differential leucocyte counts, offspring larva swimming activity and global DNA methylation in offspring embryos. No significant effects were observed in the parental generation. The offspring were either fed the same diets as those fed to their parents (Bt–Bt or nBt–nBt) or switched from the Bt diet to the nBt diet (Bt–nBt). The Bt–Bt offspring exhibited a significantly higher body mass increase, specific growth rate and feed utilisation than fish fed the nBt–nBt diet and/or fish fed the Bt–nBt diet. Liver and mid-intestinal gene transcript levels of CuZn SOD were significantly higher in fish fed the nBt–nBt diet than in those fed the Bt–Bt diet. Liver gene transcript levels of caspase 6 were significantly lower for the nBt–nBt group than for the Bt–Bt group. Overall, enhanced growth performance was observed in fish fed the Bt diet for two generations than in those fed the nBt diet for one and two generations. Effects observed on gene biomarkers for oxidative stress and the cell cycle (apoptosis) may be related to the contamination of nBt-maize with fumonisin B1 and aflatoxin B1. In conclusion, it is suggested that Bt-maize is as safe and nutritious as its nBt control when fed to zebrafish for two generations.

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.

To investigate effects on glucose and amino acid (AA) metabolism and neuroendocrine regulation of energy homoeostasis and GH-IGF system growth regulation, Atlantic salmon were fed high levels of plant protein (PP) and vegetable oil (VO) for 1 year and compared to fish fed intermediate levels or a control diet based on fish meal and fish oil. All plant-fed groups had reduced liver glucokinase mRNA and increased muscle glycogen compared to the control group. As digestible starch was highest in the control diet, the increased muscle glycogen might be related to other factors such as lower levels of lysine or higher levels of glucogenic AAs. There were minor effects on transcription of genes related to leptin, ghrelin and GH-IGF system. However, muscle growth hormone receptor was elevated in fish fed high PP, probably related to lipolytic action in these fish. Fish fed high levels of both PP and VO simultaneously had elevated plasma alanine aminotransferase and glucose and of pentose phosphate pathway enzymes. Furthermore, this group had elevated concentrations of some free AAs, especially proline. These results support interaction effects when both high levels of PP and VO are used together, which needs to be considered in the formulation of aquafeeds.

Genetically modified (GM) plants were first grown commercially more than 20 years ago, but their use is still controversial in some parts of the world. Many GM plant varieties are produced in large quantities globally and are approved for use in fish feeds both in Norway and the European Union. European consumers, however, are skeptical to fish produced by means of GM feed ingredients. Concerns have been raised regarding the safety of GM plants, including potential toxicity and (or) allergenicity of the novel protein, potential unintended effects, and risk of horizontal gene transfer to other species. This review will present the current state of knowledge regarding GM plants as fish feed ingredients, focusing on fish performance and health as well as the fate of the GM DNA fragments in the fish, identifying limitations of the current work and areas where further research is needed.

The aim of this study was to examine the presence or absence of dietary transgenic (Roundup Ready® soybean – RRS®) and soybean DNA (sRubisco) in the intestinal tract of Atlantic salmon fed either genetically modified (GM) or conventional (non-GM) soybeans. Uptake of dietary DNA was evaluated in the post gastric intestine (pyloric ceca – PC, mid intestine – MI and distal intestine – DI) after continuous feeding (6 months), feed restriction and re-feeding using qPCR and in situ hybridization. No transgenic DNA fragments were detected in any of the intestinal samples using event specific primers. Soybean DNA was detected in all segments of the intestinal tissue (PC, MI and DI) and visualized in the cell vacuolar system of the DI in the apex area of the intestinal fold. Dietary DNA was gradually cleared from the intestinal tissues when feed was restricted and could not be detected after 5 days. Re-feeding resulted in dietary plant DNA uptake after 2 h. The results show that the salmon intestine is able to take up dietary plant DNA shortly after feed intake and that one of the factors affecting uptake and clearance of nucleic acids in the various intestinal segments are the feeding status of the fish.

This study was conducted to investigate potential differences between genetically modified (GM) Roundup Ready® soy and its near-isogenic maternal line as feed ingredients for Atlantic salmon, with focus on intestinal changes commonly caused by soybean meal, histomorphology of other organs and stress response. A 7-month feeding trial was conducted with an inclusion level of 25% GM soy in the diet. Samples for histology were collected after 4 months, after 6 months, when a cross-over of the diet groups was conducted, and at the end of the trial of the crossed-over groups. Histomorphology of spleen, head kidney and mid intestine exhibited no differences between the diet groups, while glycogen deposits in liver were decreased in the GM fed fish at the final sampling. Common soybean meal-induced changes of the distal intestine in Atlantic salmon were observed in both diet groups at all sampling points, within levels expected at the current inclusion level of soy in the diets. However, mucosal fold height in the distal intestine was lower in the GM fed group at one of the three sampling points, and mucosal fold fusion was more pronounced in this group overall in the trial. A stress test conducted at the end of the trial gave responses in haematological parameters, plasma nutrients and mRNA transcription of heat shock protein (HSP) 27 in both liver and distal intestine, but responses were similar between the two diet groups, indicating similar ability to handle stress. The cross-over design, implemented to look at reversibility of potential GM-effects, proved to be inadequate as the crossing of diet groups in itself caused responses that would obscure possible minor diet effects. In conclusion, minor differences were observed between the diet groups; however, GM soy did not appear to cause any adverse effects on organ morphology or stress response compared to non-GM soy.

A 20-d zebrafish (Danio rerio) feeding trial, in which a near doubling of fish weight was achieved, was conducted with GM feed ingredients to evaluate feed intake, growth, stress response and uptake of dietary DNA. A partial aim of the study was to assess zebrafish as a model organism in GM safety assessments. Roundup Ready®soya (RRS®), YieldGard®Bt maize (MON810) and their non-modified, maternal, near-isogenic lines were used in a 2 × 2 factorial design. Soya variety and maize variety were the main factors, both with two levels; non-GM and GM. Compared with fish fed non-GM maize, those fed GM maize exhibited significantly better growth, had lower mRNA transcription levels ofsuperoxide dismutase (SOD)-1and a tendency (non-significant) towards lower transcription ofheat shock protein 70in liver. Sex of the fish and soya variety had significant interaction effects on total RNA yield from the whole liver and transcription ofSOD-1, suggesting that some diet component affecting males and females differently was present in different levels in the GM and the non-GM soya used in the present study. Dietary DNA sequences were detected in all of the organs analysed, but not all of the samples. Soya and maizerubisco(non-transgenic, multicopy genes) were most frequently detected, while MON810 transgenic DNA fragments were detected in some samples and RRS®fragments were not detected. In conclusion, zebrafish shows promise as a model for this application.

The aim of this study was to investigate potential differences in liver protein expression of Atlantic salmon fed genetically modified (GM) Roundup Ready® soy at a high inclusion level (25% inclusion, constituting 21% of crude protein in the diet) for 7 months or a compositionally similar non-GM diet. The liver was selected as the target organ due to its importance in the general metabolism, and 2D gel electrophoresis used as a screening tool. Samples from 12 individual fish from each diet group were evaluated. Of a total of 781 analysed protein spots, only 36 were significantly different by ANOVA (p

Responses to GM maize Bt-maize, MON810) expressing Cry1Ab protein from the soil bacterium Bacillus thuringiensis (Bt) in diets for both normal and immune-sensitised (with soyabean meal (SBM)-induced enteropathy) post-smolt Atlantic salmon were investigated following 33 and 97 d of exposure. Triplicate tanks of salmon were fed one of four diets, all containing 20 % whole-kernel meal maize, either Bt-maize or its near-isogenic maternal line, without or with 15 % extracted SBM inclusion. The fish fed Bt-maize utilised the feed less efficiently, as revealed by lower protein and mineral digestibilities and lower lipid and energy retention efficiencies. Higher intestinal weight, as well as increased interferon-γ and decreased sodium–glucose co-transporter mRNA expression, and a transient increase in T-helper cell presence, as measured by cluster of differentiation 4 (CD4) protein in the distal intestine (DI), may partly explain the lower nutrient digestibilities and retentions. The Bt-maize seemed to potentiate oxidative cellular stress in the DI of immune-sensitised fish, as indicated by increases in superoxide dismutase and heat shock protein 70 mRNA expression. The data suggest that Cry1Ab protein or other antigens in Bt-maize have local immunogenic effects in salmon DI. No systemic immune responses could be detected, as indicated by haematology, differential leucocyte counts, plasma clinical chemistry, as well as absence of Cry1Ab-specific antibodies and Cry1Ab protein in plasma. The responses to Bt-maize observed in the present study differed from results from earlier studies in salmon and other animals fed the same event Bt-maize. Longer-term experiments and more in-depth studies on intestinal physiology and immune responses are needed to evaluate health implications.

The effects of feeding six diets spiked with increasing levels of DON for 45 days to zebrafish (Danio rerio) on performance and liver gene biomarkers were investigated. In addition long term effects on fecundity, offspring larvae swimming activity and global DNA methylation in embryos were investigated. Zebrafish performance was not affected. Liver CYP1A mRNA levels were significantly higher in fish fed 2.0 ppm DON compared to the control group, 0.1, 0.5 and 1.5 ppm group. Gene transcripts of CuZn SOD and Cyclin G1 increased with increasing content of dietary DON. The percentage of 5-methylcytosine in embryos did not differ and was 7.0-7.1% across the groups. Fecundity showed a biphasic response pattern. Interestingly, fish fed 1.5 ppm DON had 22% higher fecundity compared to control. A trend towards increased larvae swimming activity was seen in the high DON group. Our data suggest that DON is detoxified in the liver through the phase 1 system resulting in a disturbance in the oxidative balance. We do not know if effects observed on fecundity and larvae swimming activity are attributed to a direct interaction of DON with the reproductive organ or secondary to the maternal/paternal liver oxidative imbalance.

The present study was conducted to follow up on apparent differences in growth, relative organ sizes, cellular stress and immune function in Atlantic salmon fed feed containing GMBacillus thuringiensismaize compared with feed containing the non-modified parental maize line. Gene expression profiling on the distal intestinal segment and liver was performed by microarray, and selected genes were followed up by quantitative PCR (qPCR). In the liver, qPCR revealed some differentially regulated genes, including up-regulation of gelsolin precursor, down-regulation of ferritin heavy subunit and a tendency towards down-regulation of metallothionein (MT)-B. This, combined with the up-regulation of anti-apoptotic protein NR13 and similar tendencies for ferritin heavy chain and MT-A and -B in the distal intestine, suggests changes in cellular stress/antioxidant status. This corresponds well with and strengthens previous findings in these fish. To exclude possible confounding factors, the maize ingredients were analysed for mycotoxins and metabolites. The GM maize contained 90 μg/kg of deoxynivalenol (DON), while the non-GM maize was below the detection limit. Differences were also observed in the metabolite profiles of the two maize varieties, some of which seemed connected to the mycotoxin level. The effects on salmon observed in the present and previous studies correspond relatively well with the effects of DON as reported in the literature for other production animals, but knowledge regarding effects and harmful dose levels in fish is scarce. Thus, it is difficult to conclude whether the observed effects are caused by the DON level or by some other aspect of the GM maize ingredient.