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Systematic granulomatosis is a chronic disease that affects the majority of farmed meagre (A. regius). Nutritional imbalances and overcrowding can increase the risk to suffer oxidative stress, and consequently impact the incidence of granulomatosis. In order to evaluate this, juvenile meagre were fed five isolipidic (16.7 %) and isoproteic (49.6 %) fish meal and fish oil-based feeds prepared by adding different levels of vitamin C, minerals (Mn, Zn, Se) with constant vitamin E and K (300 and 35 mg kg-1, respectively): Diet KEC (100 mg kg-1C), Diet KEC+Mn/Zn/Se (100 mg kg-1C, 40 mg kg-1Mn, 200 mg kg-1Zn, 1.5 mg kg-1Se), Diet KECC (600 mg kg-1C), Diet KECCC (1,200 mg kg-1C), Diet KECCCC (3,200 mg kg-1C). All diets were tested at 3.20 kg m-3, but diets KECC and KECCCC was also tested at 6.20 kg m-3. Growth performance was only affected by stocking density, being lower at high density. Percentage of fish with granulomas was significantly lower in fish fed with the highest dietary vitamin C contents (KECCC and KECCCC) at low density. TBARS content was correlated with the percentage of granulomas in liver (R2=0.9439, y=0.003x-0.1242) denoting the involvement of an imbalance oxidative status in the appearance of granulomas. The present results show that high levels of vitamin C (1,200-3,200 mg kg-1C) and low stock density (3.20 kg m-3) favours the growth of juvenile meagre, reducing the lipid peroxidation indicators and decreasing the incidence of granulomas, which confirms that this pathology is mostly triggered by deficiency of antioxidant nutrients, particularly vitamin C.

Aquaculture, the farming of fish and seafood, is recognized as a highly efficient system for producing protein for human consumption. In contrast, many terrestrial animal protein production systems are inefficient, impacting land use and exacerbating climate change. Humankind needs to adopt a more plant-centric diet, the only exception being fish consumed as both a source of protein and essential dietary nutrients such as omega-3 fatty acids. Here we consider the implications of such a transition, and the challenges that aquaculture must overcome to increase productivity within planetary boundaries. We consider how agriculture, specifically crops, can provide solutions for aquaculture, especially the sectors that are dependent on marine ingredients. For example, agriculture can provide experience with managing monocultures and new technologies such as genetically modified crops tailored specifically for use in aquaculture. We propose that a closer connection between agriculture and aquaculture will create a resilient food system capable of meeting increasing dietary and nutritional demands without exhausting planetary resources. Aquaculture must develop within planetary boundaries. Experience from agriculture, such as in managing monocultures and using genetically modified crops, can inform sustainable solutions for aquaculture.

An 8-week feeding trial was conducted to evaluate the effects of dietary n-3 LC-PUFA levels on growth performance, tissue fatty acid profiles and relative expression of genes involved in the lipid metabolism of mud crab (Scylla paramamosain). Ten isonitrogenous diets were formulated to contain five n-3 LC-PUFA levels at 7 and 12 % dietary lipid levels. The highest weight gain and specific growth rate were observed in crabs fed the diets with 19·8 and 13·2 mg/g n-3 LC-PUFA at 7 and 12 % lipid, respectively. Moisture and lipid contents in hepatopancreas and muscle were significantly influenced by dietary n-3 LC-PUFA at the two lipid levels. The DHA, EPA, n-3 LC-PUFA contents and n-3:n-6 PUFA ratio in hepatopancreas and muscle significantly increased as dietary n-3 LC-PUFA levels increased at both lipid levels. The expression levels of -6 fatty acyl desaturase and acyl-CoA oxidase in hepatopancreas increased significantly, and expression levels of fatty acid synthase, carnitine palmitoyltransferase I and hormone-sensitive TAG lipase were down-regulated, with increased dietary n-3 LC-PUFA regardless of lipid level. Based on weight gain, n-3 LC-PUFA requirements of S. paramamosain were estimated to be 20·1 and 12·7 mg/g of diet at 7 and 12 % dietary lipid, respectively. Overall, dietary lipid level influenced lipid metabolism, and purified, high-lipid diets rich in palmitic acid reduced the n-3 LC-PUFA requirement of juvenile mud crab.

Lipidomics methodologies traditionally utilize either reverse phase- or hydrophilic interaction liquid chromatography-type separations; however, supercritical fluid chromatography can offer a rapid normal phase type separation while reducing the dependence on organic solvents. However, normal phase type lipid separations typically lack pronounced intraclass separation, which is problematic for complex lipidomes containing very-long-chain polyunsaturated fatty acids, especially those from genetically modified organisms. A high-strength silica C18 method was developed, which benefitted from discrete class separation, as well as displaying intraclass selectivity sufficient for profiling flesh of salmon fed with a diet supplemented with oil from the genetically engineered oilseed Camelina sativa, a terrestrial oilseed with a fish oil-type profile. Salmon fed a diet containing this Camelina oil were found to have flesh enriched in triacylglycerols and phospholipids containing 18:3, 20:5, and 22:6, whereas salmon fed the control diet were differentiated by shorter chain plant-type fatty acids integrated within complex lipids. Coupled with active scanning quadrupole technology, data acquisition was enhanced, allowing for fragmentation data to be acquired in a data independent fashion, permitting acyl chain identification of resolved isomers. Therefore, we have developed a method, which is amenable for lipidomics studies of complex lipidomes, specifically those altered by synthetic biology approaches.

Ensuring lumpfish health and welfare in salmon farms is vital to reduce the high mortality rates reported and to guarantee a high delousing efficiency. Recent observations of farmed lumpfish livers have shown colours ranging from pale (colours 1 and 2), through bright orange (colours 3 and 4), to dark reddish-brown (colours 5 and 6), some of which may be related to welfare condition. To characterize the status of lumpfish deployed in four Faroese salmon farms, several welfare indicators were assessed: a weight-length relationship, scoring of external physical damage, and after dissection, stomach content and liver colour scoring. Liver samples were weighed, stored and analysed for lipid content, lipid classes, total pigments, fatty acid profile and histopathology to explain the differences between the mentioned liver colours. Bright orange livers, liver colours 3 and 4, were related to increased levels of carotenoid pigments rather than levels of lipids and appear to reflect good fish welfare. However, dark reddish-brown colours, liver colours 5 and 6, were associated with very low levels of triacyl glycerides in the liver, indicating use of lipid reserves and poor welfare condition. Histopathology confirmed that the dark reddish-brown livers, liver colours 5 and 6, formed a distinct group. Thus, liver colour was shown to be a good welfare indicator and should be monitored in farms.

The use of low fishmeal/fish oil in marine fish diets affects dietary essential fatty acids (EFAs) composition and concentration and, subsequently, may produce a marginal deficiency of those fatty acids with a direct impact on the fish intestinal physiology. Supplementation of essential fatty acids is necessary to cover the requirements of the different EFAs, including the ones belonging to the n-6 series, such as arachidonic acid (ARA). ARA, besides its structural role in the configuration of the lipid classes of the intestine, plays an important role in the functionality of the gut-associated immune tissue (GALT). The present study aimed to test five levels of dietary ARA (ARA0.5 (0.5%), ARA1 (1%), ARA2 (2%), ARA4 (4%), and ARA6 (6%)) for European sea bass (Dicentrarchus labrax) juveniles in order to determine (a) its effect in selected distal intestine (DI) lipid classes composition and (b) how these changes affected gut bacterial translocation rates and selected GALT-related gene expression pre and post challenge. No differences were found between distal intestines of fish fed with the graded ARA levels in total neutral lipids and total polar lipids. However, DI of fish fed with the ARA6 diet presented a higher (P

Untargeted lipidomic analysis was conducted to explore how different dietary docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) ratio and, specifically, how an optimal ratio (2.3) compared to a suboptimum ratio (0.6) impacted lipid molecular species and the positional distribution of fatty acids in hepatopancreas of mud crab. The results indicated that major category of lipid affected by dietary DHA/EPA ratio was glycerophospholipids (GPs). The optimum dietary DHA/EPA ratio increased the contents of DHA bound to the sn-2 and sn-3 positions of phosphatidylcholine (PC) and triacylglycerol, EPA bound to the sn-2 position of phosphatidylcholine and 18:2n-6 bound to the sn-2 position of phosphatidylethanolamine (PE). Increased dietary DHA/EPA ratio also led to competition between arachidonic acid (ARA) and 18:2n-6 bound to esterified sites. Appropriate dietary DHA/EPA ratio can not only improve the growth performance and nutritional quality of mud crab, but also provide higher quality products for human consumers.

The regulation of lipogenesis and lipolysis mechanisms related to consumption of lipid has not been studied in swimming crab. The aims of present study were to evaluate the effects of dietary lipid levels on growth, enzymes activities, and expression of genes of lipid metabolism in hepatopancreas of juvenile swimming crab. Three isonitrogenous diets were formulated to contain crude lipid levels at 5.8 %, 9.9 % and 15.1 %, respectively. Crabs fed the diet containing 15.1 % lipid had significantly lower weight gain, specific growth rate and survival, and higher feed conversion ratio than those fed the 5.8 % and 9.9 % lipid diets. Crabs fed 5.8 % lipid had lower malondialdehyde concentrations in the hemolymph and hepatopancreas than those fed the other diets. Highest glutathione peroxidase in hemolymph and superoxide dismutase in hepatopancreas were observed in crabs fed 5.8 % lipid. The lowest fatty acid synthase and glucose 6-phosphate dehydrogenase activities in hepatopancreas were observed in crabs fed 15.1 % lipid, whereas crabs fed 5.8 % lipid had lower carnitine palmitoyltransferase-1 activity than those fed the other diets. Crabs fed 15.1 % lipid showed lower hepatopancreas expression of genes involved in LC-PUFA biosynthesis, lipoprotein clearance, fatty acid uptake, fatty acid oxidation, lipid anabolism and lipid catabolism than those fed the other diets, whereas expression of some genes of lipoprotein assembly and fatty acid oxidation were up-regulated compared with crabs fed 5.8 % lipid. Overall, high dietary lipid level can inhibit growth, reduce feed utilization and reduce antioxidant enzyme activities. Moreover, dietary lipid influenced enzyme activities and gene expression involved in lipid metabolism of juvenile swimming crab., The present study was supported by National Key R & D Program of China (2018YFD0900400), China Agriculture Research System (CARS-48), National Natural Science Foundation of China (41476125), Nature Science Foundation of Zhejiang Province (LY17C190002), Key Research Program of Zhejiang Province of China (2018C02037), Zhejiang Aquaculture Nutrition & Feed Technology Service Team (ZJANFTST2017-2) and Major Agriculture Program of Ningbo (2017C110007). This research was also sponsored by the K. C. Wong Magna Fund in Ningbo University.

The long-chain (≥C20) polyunsaturated fatty acid biosynthesis capacity of fish varies among species, with trophic level hypothesised as a major factor. The biosynthesis capacity is largely dependent upon the presence of functionally diversified fatty acyl desaturase 2 (Fads2) enzymes, since many teleosts have lost the gene encoding a Δ5 desaturase (Fads1). The present study aimed to characterise Fads2 from four teleosts occupying different trophic levels, namely Sarpa salpa, Chelon labrosus, Pegusa lascaris and Atherina presbyter, which were selected based on available data on functions of Fads2 from closely related species. Therefore, we had insight into the variability of Fads2 within the same phylogenetic group. Our results showed that Fads2 from S. salpa and C. labrosus were both Δ6 desaturases with further Δ8 activity while P. lascaris and A. presbyter Fads2 showed Δ4 activity. Fads2 activities of herbivorous S. salpa are consistent with those reported for carnivorous Sparidae species. The results suggested that trophic level might not directly drive diversification of teleost Fads2 as initially hypothesised, and other factors such as the species' phylogeny appeared to be more influential. In agreement, Fads2 activities from P. lascaris and A. presbyter were similar to their corresponding phylogenetic counterparts Solea senegalensis and Chirostoma estor., Te present study was funded by Ministerio de Economía y Competitividad (AGL2015-70994-R). Dr Covadonga Rodríguez is a member of the Instituto de Biotecnología de Canarias (ITB).

Systemic granulomatosis has already been reported in meagre larvae with an adequate feeding protocol and enrichment media preventing its appearance in the first weeks of life. Afterwards, the control of this disease could be prevented through nutritional components of the inert food, being the antioxidants the key to success. For this reason, in the present study, meagre larvae were reared from 30 days post hatching (dph) with five isonitrogenous and isolipidic experimental microdiets with different levels of vitamin E and C: C– (40 mg kg−1 E, 100 mg kg−1C), C+ (400 mg kg−1 E, 1000 mg kg−1C), Krill (400 mg kg−1 E, 1000 mg kg−1C and substitution of fish oil by krill oil), EC (200 mg kg−1 E, 500 mg kg−1C) and EECC (800 mg kg−1 E, 2000 mg kg−1C). Prior to this, larvae were co-fed with rotifers and Artemia following a protocol which prevented the appearance of granulomas, as previously demonstrated. The substitution of fish oil by krill oil significantly increased levels of eicosapentaenoic acid (EPA, 16.6%) and docosahexaenoic acid (DHA, 17.6%) in meagre, consequently increasing the peroxidation index, which in turn translated into a higher incidence of granulomas. Although even low levels of vitamin E and C (40 mg kg−1 E, 100 mg kg−1C; C-) allowed the adequate growth of larvae, these levels were not enough to prevent the appearance of granulomas, requiring superior levels of both antioxidant vitamins (800 mg kg−1 E and 2000 mg kg−1C) to mitigate systemic granulomatosis. This mitigation was simultaneous with the reduction of thiobarbituric acid reactive substances TBARs content in larvae, which were highly correlated with the appearance of granulomas (R2 = 0.892, y = 0.0446× + 0.0756). A strong negative correlation was observed between the dietary levels of vitamin E (y = −0.0098× + 11.174, R2 = 0.8766, p value = .019, r = −0.93) and vitamin C (y = −0.0022× + 6.4777, R2 = 0.9278, p value = .003, r = −0.96) and the percentage of larvae with granulomas. The results showed that the occurrence of systemic granulomatosis seems to be associated to the larvae peroxidation status, so that high dietary levels of vitamin E and C (800 and 2000 mg kg−1, respectively; Diet EECC), reduced lipid peroxidation and completely prevented the appearance of granulomas in meagre larvae at 44 dph.

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.

Copper is an essential micronutrient for living organisms, but its biological function has not been sufficiently studied, especially in crustaceans. Therefore, the present study aimed to evaluate the role of copper in the maintenance of energy homeostasis by mediating lipid and energy metabolism in Litopenaeus vannamei. L. vannamei (initial weight 0.90 ± 0.00 g) were fed diets containing 12.4 (Cu12.4), 22.0 (Cu22.0), 31.0 (Cu31.0), 41.7 (Cu41.7) and 49.8 (Cu49.8) mg kg−1 Cu for 8 weeks. Lipid and energy metabolism-related biochemical and molecular parameters were determined, and ultrastructural observation performed in hepatopancreas. Growth performance, tissue Cu deposition, antioxidant parameters, and expression of genes involved in Cu homeostasis and autophagy were also evaluated. Dietary copper supplementation improved growth, Cu concentration in tissue, and antioxidant capacity compared to the Cu-deficient diet. Copper promoted lipolysis and β-oxidation, confirmed by elevated expression levels of cpt1, ampkγ, acbp, acadl, acadm, acadvl, acox1, acox3 and decreased expression of srebp, acc1, 6pgd, and markedly increased activities of LPS and CPT1 and decreased ACC activity. Expression levels of genes related to autophagy (atg13, atg3, atg12, atg9, beclin1) were up-regulated, and autolysosomes and autophagosome were observed in the hepatopancreas of shrimp fed diets supplemented with copper. Furthermore, energy metabolism related parameters (ATPase, cytochrome c oxidase, citrate synthase, adenosine 5′-triphosphate) and genes involved in tricarboxylic-acid cycle (cs, aco, idh, ogdh, sdhc, sdhd, fh, mdh) were up-regulated in shrimp fed copper supplemented diets. The present study showed that copper-mediated lipolysis and autophagy can coordinate energy generation, and provided new insight into copper nutrition and the potential beneficial effects of dietary Cu on energy metabolism in crustaceans.

All living organisms require copper for growth and development, but the gene expression profiles and molecular mechanisms underpinning dietary copper are poorly investigated. Therefore, the present study aimed to determine the potential metabolic changes in response to dietary copper based on analysis of hepatopancreas transcriptome in Litopenaeus vannamei. Three practical diets were formulated to supplement 0 (control diet; Csingle bondCu) and 40 mg kg−1 inorganic Cu (CuSO4·5H2O; Isingle bondCu) and copper amino acid chelate (Osingle bondCu), with analyzed Cu being 12.4, 49.8 and 50.0 mg kg−1, respectively. Shrimp fed Isingle bondCu and Osingle bondCu diets had higher percent weight gain and Cu concentration in tissues. Some essential amino acids (lysine, methionine, isoleucine, leucine, valine) and non-essential amino acids (tyrosine, glycine, aspartic acid, proline and serine) in hepatopancreas significantly increased in shrimp fed the copper supplemented diets. Transcriptome analysis indicated a total of 742 and 912 genes were differentially expressed (q < 0.001; log2fold change ≥2) in shrimp fed the Isingle bondCu and Osingle bondCu diets, respectively, in comparison to shrimp fed the control diet. Five and eight significantly changed pathways were annotated in the Csingle bondCu vs. Isingle bondCu and Csingle bondCu vs. Osingle bondCu comparisons, with metabolism the leading category for both. Similarly, the proportion of differentially expressed genes revealed that most were enriched in the category of metabolism. Further analysis revealed that dietary copper mainly affected amino acid and glycerophospholipid metabolism. Moreover, two significantly changed pathways (phagosome and IL-17 signaling pathway) related to the immune system were identified in shrimp fed the Osingle bondCu diet. The present study analyzing the hepatopancreas transcriptome identified potential roles of dietary copper on amino acid and glycerophospholipid metabolism and provided new insight that will be valuable in future studies to further elucidate the nutritional molecular basis of copper.

The digestive system presents daily rhythms at both physiological and histological levels. Although cell morphology rhythms in mammals have been reported, they have scarcely been investigated in fish. The aim of the present research was to investigate the existence of daily rhythms in the morphology of cells in the liver and intestine of a teleost fish, the European sea bass (Dicentrarchus labrax), and how feeding time influences them. Regarding liver, we also focused on differences between the two metabolic zones: perivenous and periportal. For this purpose, fish were divided into two groups: fish fed once a day in the mid-light phase (ML) or the mid-dark phase (MD). After 1 month under each feeding regime, liver and intestine samples were collected every 4 h during a 24-h cycle, and different parameters were studied by light microscopy and image analysis. Daily rhythms occurred in most of the parameters evaluated in the liver. The effect of feeding time depended on the metabolic zone: the rhythms in the periportal zone were synchronized mainly by the light/dark cycle regardless of feeding time, whereas in the perivenous zone, rhythms were influenced more by feeding time. In the intestine, a daily rhythm in villi height was found with acrophases coinciding with feeding time in each group. These findings show for the first time the existence of cellular morphological rhythms in fish liver and intestine, and highlight the interactions between light and feeding cycles in the different metabolic zones of the liver.

Routine gill swabbing is a non-destructive sampling method used for the downstream qPCR detection and quantitation of the pathogen Neoparamoeba perurans, a causative agent of amoebic gill disease (AGD). Three commercially available swabs were compared aiming their application for timelier AGD diagnosis (Calgiswab® (calcium alginate fibre-tipped), Isohelix® DNA buccal and cotton wool-tipped). Calcium alginate is soluble in most sodium salts, which potentially allows the total recovery of biological material, hence a better extraction of target organisms' DNA. Thus, this study consisted of (a) an in vitro assessment involving spiking of the swabs with known amounts of amoebae and additional assessment of retrieval efficiency of amoebae from agar plates; (b) in vivo testing by swabbing of gill arches (second, third and fourth) of AGD-infected fish. Both in vitro and in vivo experiments identified an enhanced amoeba retrieval with Calgiswab® and Isohelix® swabs in comparison with cotton swabs. Additionally, the third and fourth gill arches presented significantly higher amoebic loads compared to the second gill arch. Results suggest that limiting routine gill swabbing to one or two arches, instead of all, could likely lead to reduced stress-related effects incurred by handling and sampling and a timelier diagnosis of AGD.

The present study evaluated the effects of dietary Zn level on growth performance, serum and hepatopancreas metabolites, expression of genes involved in lipid and energy metabolism, and the signalling pathway of dietary Zn-induced lipolysis. Five isonitrogenous and isolipidic diets were formulated to contain different Zn levels: 46·4 (basal diet), 77·2, 87·0, 117·1 and 136·8 mg/kg, respectively. The results indicated that shrimp fed the diet containing Zn at 117·1 mg/kg had higher weight gain and specific growth rate, and the lowest feed intake and feed conversion rate, than shrimp fed the other diets. The deposition rate of Zn in whole body significantly decreased with increasing dietary Zn level. Dietary Zn prevented the accumulation of free radicals and improved antioxidant activities by increasing Cu/Zn superoxide dismutase and reducing malondialdehyde in hepatopancreas. Dietary Zn supplementation enhanced lipase activity and adiponectin, which could promote TAG breakdown and fatty acid oxidation and lead to reduced lipid in hepatopancreas. The mRNA expressions ofob-rb,adipor,camkkβ,ampk,cd36,mcdandcpt1involved in Zn-induced lipid catabolism were up-regulated, and the expressions ofsrebp,acc,fasandscd1were down-regulated. The mRNA levels of SLC39 family genes (zip3,zip9,zip11andzip14) in hepatopancreas were up-regulated with increasing dietary Zn level. The results demonstrated that dietary Zn level could significantly affect growth performance, tissue deposition of Zn, lipid metabolites and expression of genes involved in lipogenesis and lipolysis inLitopenaeus vannamei.

This study investigated whether dietary supplementation of phytogenic compounds limonene and thymol had synergistic or additive effects on growth and selected nutritional physiology pathways in Nile tilapia. A 63-day feeding experiment was conducted using fish of 1.5 ± 0.0 g (± standard error) fed on a commercial diet coated with either 0 ppm limonene and thymol (control), 400 ppm limonene (L), 500 ppm thymol, (T) or a combination of 400 ppm limonene and 500 ppm thymol (LT). Final fish weight (FW) was significantly improved to similar extents by diet LT (16.7 ± 0.3 g) and L (16.6 ± 0.4 g). Dietary thymol alone and the control did not enhance FW (15.0 ± 0.4 g and 13.7 ± 0.4 g respectively). Dietary thymol had shown a strong tendency to improve somatic growth (P = .052). The analysed candidate genes involved in the pathways of nutrient digestion, absorption and transport (muc), lipid metabolism (lpl), antioxidant enzymes (cat) and somatotropic axis growth (igfI) were also up-regulated to similar extents in Nile tilapia by diet L and LT (P < .05), above the regulation observed with the diet supplemented exclusively with thymol. This suggests lack of synergistic or additive effects on growth and nutritional physiology pathways when limonene and thymol are supplied in the diet., This work was funded by the Schlumberger Foundation PhD Training Program, and the Government of Uganda through the Agricultural Technology and Agribusiness Services (ATAAS) Project of the National Agricultural Research Organization (NARO).

Carps, barbels and other cyprinids are the major contributors to freshwater aquaculture at global scale. Nevertheless, freshwater fish aquaculture needs to diversify their production in order to offer consumers new species. Tench (Tinca tinca) is a freshwater species with great interest for the diversification of continental aquaculture. However, up to date, no commercial formulated diet exists for this species in order to optimize their nutritional requirements and the quality of its final product. Using multiple methodological approaches, the aim of this study was to evaluate the long chain polyunsaturated fatty acid (LC-PUFA) metabolism of T. tinca. Firstly, the molecular cloning and functional characterisation by heterologous expression in yeast of a desaturase (Fads2) and two elongases (Elovl2 and Elovl5) involved in LC-PUFA biosynthesis, and the analysis of gene expression among tissues were performed. Secondly, in order to confirm the LC-PUFA biosynthesis capacity of isolated hepatocytes and enterocytes, cells were incubated with [1-14C] labelled linoleic acid (18:2n-6, LA), linolenic acid (18:3n-3, ALA) and eicosapentaenoic acid (20:5n-3, EPA). In yeast, Fads2 showed a Δ6/Δ5 bifunctional activity. Elovl2 was more active over C20 and C22 substrates, whereas Elovl5 was over C18 and C20. Liver displayed the highest expression for the three target genes (fads2, elovl2 and elovl5). Incubated cells also showed Fads2 bifunctional activity as well as elongation products in concordance with yeast heterologous expression results. Importantly, our results demonstrated that tench is able to biosynthesise docosahexaenoic acid (DHA) from 18:3n-3 in both hepatocytes and enterocytes, a capacity that seems to explain in part the surprisingly high levels of DHA found in the fish flesh compared to its dietary supply. Tench is a promising freshwater species with a potential capacity to endogenously increase its flesh DHA contents, reducing the impact that the usage of fish oils from forage fisheries may have on the aquaculture industry., This study was funded by Ministerio de Economía y Competitividad (AGL2015-70994-R).

Atlantic bluefin tuna (ABT; Thunnus thynnus) is a highly regarded and consumed species, but farming is still in its infancy. Currently, nothing is known about the presence of circadian rhythmicity at central or peripheral tissues, or if there are daily rhythms in expression of genes involved in lipid metabolism. In order to elucidate clock gene regulation of genes of lipid metabolism in ABT, six clock genes (bmal1, clock, cry1, cry2, per1 and per2) were sequenced and 24 h expression of these genes determined in brain and liver of fish acclimated to a light:dark (L:D) photoperiod. Additionally, the daily expression of lipid metabolism and digestive enzyme genes in ABT was also determined in liver. All six clock genes displayed rhythmicity in the brain and liver, other than cry2, which did not show an acrophase in liver. In liver, all the transcription factors analysed other than srebp1 and srebp2 displayed rhythmicity, with lxr and pparα displaying diurnal expression, whereas pparγ was highly expressed at the end of the scotophase. Some of the target genes of lxr such as elovl5 and lpl also oscillated rhythmically, with acrophases during the photophase. In contrast, only three of the eight digestive enzyme genes studied displayed rhythmicity, at different times of the day, suggesting that either ABT display different feeding periods or the digestion of some nutrients (e.g. lipids) is prioritized over others. The present study showed that clock and lipid metabolism genes displayed strong daily rhythmicity in ABT brain and liver, which could be an area of considerable interest for the establishment of efficient feeding protocols in this new aquaculture species., SI

The use of plant-based feeds warrants the supplementation with selenium (Se) to cover the requirement for Atlantic salmon. Depending on its chemical form, Se is a trace element with a narrow range between requirement and toxicity for most vertebrates. Information on safe upper limit for Atlantic salmon feed supplementation is lacking. Atlantic salmon (147 g) were fed a low natural background organic Se diet (0.45 mg Se kg−1, wet weight (ww)) fortified with 5 graded levels of inorganic sodium selenite (0.45, 5.4, 11.0, 29.4, or 60.0 mg kg−1 ww) or organic selenomethionine (SeMet) (0.45, 6.2, 16.2, 21, or 39 mg kg−1 ww), in triplicate for 3 months. Excess Se supplementation was assessed by targeted biomarkers of Se toxicity pathways (e.g. markers of oxidative stress and lipid metabolism), as well as general adverse effect parameters (plasma biochemistry, hematology, liver histopathology, and growth). Safe limits were set by model-fitting the effect data in a dose-response (lower bound) bench mark dose (BMDL) evaluation. Fish fed the two highest selenite levels showed mortality while fish fed SeMet had no mortality. Fish fed 5.4–11 mg selenite kg−1 feed showed significantly (ANOVA, Tukey's t-test, p

Phytogenic compounds such as limonene and thymol have been shown to have growth-promoting properties in farmed animals but studies in fish are scarce. Two experiments (Experiments I and II) were carried out to investigate the individual effects of dietary limonene and thymol on the growth and nutritional physiology of juvenile Nile tilapia (Oreochromis niloticus). In Experiment I, the fish were fed on a commercial diet coated with limonene at 0 (Control), 200, 400, and 600 mg kg−1 (ppm), while in Experiment II thymol was supplemented in the diet at 0 (Control), 250 and 500 ppm. Our results showed a significant increase in fish weight and weight gain with diets supplemented with 400 and 600 ppm limonene compared to the control. Moreover, the expression of insulin growth factor I (igf-I), mucin-like protein (muc), oligo-peptide transporter I (pept1), lipoprotein lipase (lpl), alkaline phosphatase (alp) and catalase (cat) was up-regulated by dietary limonene. Our results confirm that dietary limonene can enhance the growth of Nile tilapia juveniles through the activation of key genes involved in somatotropic axis-mediated growth, nutrient digestion and antioxidant enzyme defence. Dietary thymol did not seem to influence growth or regulate the same pathways activated by limonene in Nile tilapia juveniles at inclusion levels up to 500 ppm. Overall, the present results suggest that potential growth-promoting effects are dependent upon the phytogenetic itself and its inclusion level.

Five isoproteic (54.8%) and isolipidic (24.1%) microdiets, which varied in their docosahexaenoic acid (DHA) content (0.25%, 0.75%, 1.64%, 1.99% and 3.17%; dw), were manufactured to determine its effects on longfin yellowtail Seriola rivoliana larvae in terms of fish biological performance, whole body fatty acid profile and incidence of skeletal anomalies from 30 dah (11.31 ± 1.79 Total Length, TL) to 50 dah (19.80 ± 0.58 mm TL). The inclusion of dietary DHA up to 3.17% (dw) improved larval resistance to air exposure, although DHA did not significantly affect fish final growth or final survival. Indeed, high levels of dietary DHA (1.99% and 3.17%, dw) tended to increase the incidence of skeletal anomalies in S. rivoliana larvae, albeit no significant differences were observed. Furthermore, the occurrence of severe anomalies such as kyphosis and lordosis, was mainly associated to the larvae fed the highest levels of dietary DHA. In terms of survival, increasing dietary DHA levels did not significantly affect longfin yellowtail survival rate, despite a tendency for enhanced survival. The results of the present study proved that the inclusion of dietary DHA in inert diets up to a 3.17% (dw) and a DHA/EPA ratio above 3.1 increased the final survival and stress resistance in S. rivoliana larvae.

While chromium (Cr) has been recognized as an essential nutrient for all animals, and dietary supplementation can be beneficial, it can also be toxic. The present study aimed to investigate the contrasting effects of dietary chromium in Pacific white shrimp Litopenaeus vannamei. Five experimental diets were formulated to contain Cr at levels of 0.82 (Cr0.82, unsupplemented diet), 1.01 (Cr1.01), 1.22 (Cu1.22), 1.43 (Cr1.43) and 1.63 (Cr1.63) mg/kg and were fed to shrimp for 8 weeks. Highest weight gain was recorded in shrimp fed the diet containing 1.22 mg/kg Cr. Shrimp fed the diet containing the highest level of Cr (1.63 mg/kg) showed the lowest weight gain and clear signs of oxidative stress and apoptosis as evidenced by higher levels of H2O2, malondialdehyde and 8-hydroxydeoxyguanosine, and expression of caspase 2, 3, 5, and lower contents of total and oxidized glutathione, and expression of Cu/Zn sod, cat, gpx, mt, bcl2. Chromium supplementation promoted glycolysis and inhibited gluconeogenesis as shown by increased activities of hexokinase, phosphofructokinase and pyruvate kinase, and reduced activity of phosphoenolpyruvate carboxykinase in shrimp fed the diet containing 1.43 mg/kg Cr. Shrimp fed the diet with 1.63 mg/kg Cr had lowest contents of crustacean hyperglycemic hormone and insulin like peptide in hemolymph. Expression of genes involved in insulin signaling pathway and glycose metabolism including insr, irs1, pik3ca, pdpk1, akt, acc1, gys, glut1, pk, hk were up-regulated, and foxO1, gsk-3β, g6pc, pepck were down-regulated in shrimp fed the diets supplemented with Cr. This study demonstrated that optimum dietary supplementation of Cr had beneficial effects on glucose homeostasis and growth, whereas excess caused oxidative damage and impaired growth. The results contribute to our understanding of the biological functions of chromium in shrimp.

Several causative factors have been proposed for the occurrence of skeletal anomalies in fish larvae, among which we quote nutritional factors, such as LC-PUFAs. This study aimed to investigate the effect of different dietary DHA and ARA level and ratio on pikeperch (Sander lucioperca) larval development and performance, digestive capacity, fatty acids composition, skeleton anomalies and molecular markers of oxidative stress status (sod, gpx, and cat), stress response (StAR, gr, pparα, hsl and pepck), fatty acid synthesis (fadsd6, elovl5), eicosanoids synthesis (pla2, cox2, lox5, pge2, and lta4h), and bone development (twist, mef2c, sox9, and alp). Pikeperch larvae were fed six microdiets containing two different dietary levels of DHA (0.5% and 3.5%) combined with three levels of ARA (1.2%, 0.6%, and 0.3%). Dietary fatty acid changes did not affect growth performance but significantly influenced enzymatic activities. A significant increase in skeletal anomalies with DHA intake increment was recorded. StAR, cox2, pla2 and hsl expression were significantly depressed in 2.5% DHA larvae. An opposite effect of dietary DHA elevation was recorded in gpx expression. Both DHA and ARA had a significant effect on pparα, gr, and pge2 expressions. Although no significant interactions were found, pge2, gr, and pparα displayed a differential pattern of expression between the different treatments. A strong association was found for the larval tissue amount of ARA and DHA with eicosanoid metabolism, stress response and skeleton anomaly related genes. These results denoted the effects of dietary LC-PUFAs on immune/stress gene regulation and their potential implication in skeleton development.

While tissue fatty acid compositions reflect that of the dietary lipid source, little information is available on how dietary oils modify lipid class and molecular species profiles in hepatopancreas of crustacean. Herein, an 8-week nutritional trial and untargeted lipidomic analysis were used to investigate the impacts of dietary n-3 PUFA lipid sources including fish oil, krill oil and linseed oil on the lipidomic characteristics of hepatopancreas of swimming crab (Portunus trituberculatus). Dietary krill oil significantly increased distribution of 20:5n-3 and 22:6n-3 at sn-2 in phosphatidylcholine and phosphatidylethanolamine compared to fish oil. Fish oil intake promoted the deposition of 20:5n-3 and 22:6n-3 at sn-1,2,3 in triglyceride compared to linseed oil, which significantly increased the specific accumulation of 18:3n-3 at sn-1,3 in triglyceride and sn-2 in phosphatidylcholine and phosphatidylethanolamine. The study revealed metabolic responses to different dietary n-3 PUFA in swimming crab, which provided novel insight into the lipid nutrition of crustacean.

The aim of this study was to evaluate the effects of increasing dietary arachidonic acid (ARA) levels (from 1 up to 6 % of total fatty acids) on European sea bass (Dicentrarchus labrax) juveniles growth performance, tissue fatty acid profile, liver morphology as well as long chain polyunsaturated fatty acids (LC-PUFA) biosynthesis, triglyceride and cholesterol synthesis and lipid transport. A diet with total fish oil (FO) replacement and defatted fish meal (FM) containing a 0.1 g ARA g-1 diet was added to the experimental design as a negative control diet. Dietary ARA inclusion levels below 0.2 g ARA g-1 diet significantly worsened growth even only 30 days after the start of the feeding trial, whereas dietary ARA had no effect on fish survival. Liver, muscle and whole body fatty acid profile mainly reflected dietary contents and ARA content increased accordingly with ARA dietary levels. Tissue eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) levels were positively correlated among them. Hepatic lipid vacuolization increased with reduced dietary ARA levels. Expressions of fatty acyl desaturase 2 and 3-hydroxy-3-methylglutaryl-coenzyme genes were up-regulated in fish fed the negative control diet compared to the rest of the dietary treatments denoting the influence of ARA on lipid metabolism. Results obtained highlight the need to include adequate n-6 levels and not only n-3 LC-PUFA levels in European sea bass diets. Keywords: Dicentrarchus labrax, arachidonic acid, growth performance, tissue fatty acid profile, lipid metabolism

Seven experimental isonitrogenous (50%) and isolipidic (16%) diets with different levels of α-tocopherol acetate (16, 100, 190, 285, 430, 880 and 1300 mg kg − 1 ) were tested during 72 days to evaluate growth performance, tissue composition, fillet oxidation and liver histology in meagre juveniles, Argyrosomus regius . Growth performance, feed conversion ratio (FCR) and tissue composition were similar among treatments ( P > 0.05). In the liver, no major differences were recorded in lipid and fatty acid composition but higher lipid vacuolization were observed in diets E100, E190 and E880. Muscle fatty acid profiles showed an increment of the highly unsaturated fatty acid (HUFA) and a decrease of the saturated fatty acid with the increase of dietary vitamin E, which was accompanied with a reduction of the muscle TBARS responses. Therefore, is suggested that diets for this species should be supplemented with 451 mg kg − 1 of DL-α-tocopherol acetate (496 UI of vitamin E), as determine by broken-line regression analysis of muscle TBARS, to provide good overall growth performance and improved fish quality and storage stability. Moreover, results suggest that vitamin E deficiency or excess may deteriorate fish health. Statement of relevance The optimization of the dietary vitamin E level will contribute to formulate a suitable diet for meagre, a candidate for European aquaculture diversification, that until now is being fed with diets specific for other species. This study will narrow the knowledge gap that exists regarding meagre nutritional needs.

Lipid sources as alternatives to fish oil could alter the nutritional value and flavor quality of crab meat affecting consumer preferences. Herein, an 8-week nutritional trial was designed to investigate the effects of dietary lipid sources including fish oil (FO), krill oil (KO), palm oil, rapeseed oil, soybean oil and linseed oil on profiles of amino acids, fatty acids and volatiles in muscle of swimming crab (Portunus trituberculatus). Volatiles of crab muscle were characterized by headspace solid-phase microextraction and gas chromatography-tandem mass spectrometry. Results revealed that crabs fed FO and KO had significantly higher levels of protein, indispensable amino acids, eicosapentaenoic acid and docosahexaenoic acid in muscle. Principal component analysis and hierarchical cluster analysis demonstrated that muscle volatiles of crabs fed different dietary oils exhibited significant variations. Dietary FO and KO significantly increased the relative levels of 3-methylbutanal, heptanal, benzaldehyde and nonanal in muscle, which may produce more pleasant flavors.

Elongation of very long-chain fatty acid 4 (Elovl4) proteins are involved in the biosynthesis of very long-chain (> C24) fatty acids and in many teleost fish species they are key enzymes in the pathway for the production of docosahexaenoic acid (DHA; 22:6n-3) from eicosapentaenoic acid (EPA; 20:5n-3). Therefore, Elovl4 may be particularly important in Atlantic bluefin tuna (ABT; Thunnus thynnus) characterised by having high DHA to EPA ratios. The present study cloned and characterised both the function and expression of an elovl4 cDNA from ABT. The Elovl4 had an open reading frame of 915 base pairs encoding a putative protein of 304 amino acids. Alignment and phylogenetic analyses indicated that the Elovl4 isoform identified in the present study was an Elovl4b. Functional characterisation demonstrated that the Elovl4b enzyme had elongase activity towards all the polyunsaturated fatty acid (PUFA) substrates assayed. The ABT Elovl4b contributed to DHA biosynthesis by elongation of EPA and DPA to 24:5n-3, the latter being desaturated to 24:6n-3 by the action of fads2 (Δ6 desaturase). Additionally, the ABT Elovl4b has a role in the biosynthesis of very long-chain PUFA up to C34, compounds of key structural roles in neural tissues such as eye and brain, which had high levels of elovl4b transcripts. Surprisingly, while the relative expression of fads2, required for the production of DHA from EPA, was increased in liver of ABT fed a diet with reduced levels of EPA and DHA, expression of elovl4b was reduced. Results indicated that ABT has enzymes necessary for endogenous production of DHA from EPA and demonstrate that Elovl4b can effectively compensate for absence of Elovl2., SI

The present study aimed to investigate whether dietary choline can regulate lipid metabolism and suppress NFκB activation and, consequently, attenuate inflammation induced by a high-fat diet in black sea bream (Acanthopagrus schlegelii). An 8-week feeding trial was conducted on fish with an initial weight of 8·16 ± 0·01 g. Five diets were formulated: control, low-fat diet (11 %); HFD, high-fat diet (17 %); and HFD supplemented with graded levels of choline (3, 6 or 12 g/kg) termed HFD + C1, HFD + C2 and HFD + C3, respectively. Dietary choline decreased lipid content in whole body and tissues. Highest TAG and cholesterol concentrations in serum and liver were recorded in fish fed the HFD. Similarly, compared with fish fed the HFD, dietary choline reduced vacuolar fat drops and ameliorated HFD-induced pathological changes in liver. Expression of genes of lipolysis pathways were up-regulated, and genes of lipogenesis down-regulated, by dietary choline compared with fish fed the HFD. Expression of nfκb and pro-inflammatory cytokines in liver and intestine was suppressed by choline supplementation, whereas expression of anti-inflammatory cytokines was promoted in fish fed choline-supplemented diets. In fish that received lipopolysaccharide to stimulate inflammatory responses, the expression of nfκb and pro-inflammatory cytokines in liver, intestine and kidney were all down-regulated by dietary choline compared with the HFD. Overall, the present study indicated that dietary choline had a lipid-lowering effect, which could protect the liver by regulating intrahepatic lipid metabolism, reducing lipid droplet accumulation and suppressing NFκB activation, consequently attenuating HFD-induced inflammation in A. schlegelii., This research was supported by the National Natural Science Foundation of China (grant no. 31802303), Natural Science Foundation of Ningbo (grant no. 2018A610343), National Key R & D Program of China (2018YFD0900400), Key Research Program of Zhejiang Province of China (no. 2018C02037), Zhejiang Aquaculture Nutrition & Feed Technology Service Team (ZJANFTST2017-2), the Open Fund of Zhejiang Provincial Top Key Discipline of Aquaculture in Ningbo University and K. C. Wong Magna Fund in Ningbo University.

Ethoxyquin (EQ; 6-Ethoxy-2,2,4-trimethyl-1,2-dihydroquinoline) has been used as an antioxidant in feed components for pets, livestock and aquaculture. However, possible risks of EQ used in aquafeed for fish health have not yet been characterized. The present study investigated the toxicity and dose-response of subchronic dietary EQ exposure at doses ranging from 41 to 9666 mg EQ/kg feed in Atlantic salmon (Salmo salar L.). Feed at concentrations higher than 1173 mg EQ/kg were rejected by the fish, resulting in reduced feed intake and growth performance. No mortality was observed in fish exposed to any of the doses. A multi-omic screening of metabolome and proteome in salmon liver indicated an effect of dietary EQ on bioenergetics pathways and hepatic redox homeostasis in fish fed concentrations above 119 mg EQ/kg feed. Increased energy expenditure associated with an upregulation of hepatic fatty acid β-oxidation and induction and carbohydrate catabolic pathways resulted in a dose-dependent depletion of intracytoplasmic lipid vacuoles in liver histological sections, decreasing whole body lipid levels and altered purine/pyrimidine metabolism. Increased GSH and TBARS in the liver indicated a state of oxidative stress, which was associated with activation of the NRF2-mediated oxidative stress response and glutathione-mediated detoxification processes. However, no oxidative DNA damage was observed. As manifestation of altered energy metabolism, the depletion of liver intracytoplasmic lipid vacuoles was considered the critical endpoint for benchmark dose assessment, and a BMDL10 of 243 mg EQ/kg feed was derived as a safe upper limit of EQ exposure in Atlantic salmon.

Taurine appears to be a crucial nutrient for teleosts, especially top predator species such as Atlantic bluefin tuna (Thunnus thynnus, L.; ABT). While dietary taurine supplementation has been highly recommended, there is a lack of studies on taurine assimilation and biosynthesis for this iconic species. The present study aims to provide insight into the molecular mechanisms involved in taurine biosynthesis and transport in ABT by studying tissue distribution and ontogenetic development of expression of cysteine dioxygenase (cdo), cysteine sulfinic acid decarboxylase (csad), 2-aminoethanethiol dioxygenase (ado) and taurine transporter (tauT) in response to graded levels of dietary taurine supplementation. The full open reading frame (ORF) for cdo and partial sequences for csad, ado and tauT were obtained, with the translated polypeptides being 202, 176, 166 and 324 amino acids, respectively. All three showed characteristics such as cupin motifs in Cdo and predicted N-glycosylation sites in Taut that are common to these genes in other species. Phylogenetic analysis showed that the ABT sequences clustered with sequences of other teleosts, and separately from mammals and molluscs. Tissue distribution varied, with adipose tissue, kidney, white muscle and testis/brain showing highest expression of cdo, csad, ado and tauT, respectively. Whole larvae expression of csad peaked at 15 dah, whereas the other genes generally increased throughout development to show highest expression at 25 dah. The nutritional trial was carried out by feeding ABT larvae from mouth opening to 14 days after hatching (dah) with rotifers (Brachionus rotundiformis) enriched with 4 different levels of taurine: 0.0 (tau0), 0.5 (tau0.5), 1.0 (tau1), and 2.0 g taurine per 106 rotifers (tau2). Rotifers effectively accumulated taurine with ABT larvae fed on treatment tau2 attaining the highest concentration of taurine. However, ABT larvae fed tau1 displayed higher growth and survival, and flexion index at 14 dah, than larvae fed the other taurine levels. Larvae fed tau1 also showed generally higher expression of tauT and cdo and digestive and antioxidant enzyme genes. While this study showed that larval ABT express taurine metabolism genes, suggesting possible synthesis that could contribute to the taurine pool in the fish, larval performance was enhanced by a level of dietary taurine (3.7 mg taurine g−1 rotifer) supplied by enrichment of rotifers at 1 g taurine per 106 rotifers., SI

Mass mortality is still one of the main constraints in larval rearing of Atlantic bluefin tuna (Thunnus thynnus L.; ABT). Early data related to the feeding sequence of ABT larvae suggested that mortality observed during the first stages of life could be partly due to nutritional deficiencies. Previous studies demonstrated that copepods ap- peared to be a superior live prey compared to rotifers during the first 2 weeks of life. Our overarching aim was to evaluate different feeding strategies during first feeding of ABT larvae from a performance, compositional and molecular perspective. In order to do so, two groups of ABT larvae were fed with either copepod (Acartia tonsa; C) nauplii or rotifers (Brachionus rotundiformis; R) enriched with Algamac 3050® from mouth opening to 13 days after hatching (dah). After this, the group C-larvae was fed either Artemia enriched with Algamac 3050® (CA), Acartia nauplii and copepodites (CC) or sea bream (Sparus aurata) yolk-sac larvae (CY), while the R group were fed on Artemia enriched with Algamac 3050® (RA) up to 18 dah. At 13 dah, larvae fed copepods (C) had grown better than those fed enriched rotifers (R) although there were no significant differences in survival. ABT larvae fed R accumulated highest eicosapentaenoate (EPA) but lowest docosahexaenoate (DHA) and total n-3 long- chain polyunsaturated fatty acids (LC-PUFA) than C-fed larvae, reflecting the dietary contents. There was no activation in the expression of the enzymes involved in EPA and DHA biosynthesis. However, the different live prey showed regulation of transcription factor, digestive enzyme, lipid metabolism and oxidative stress genes. At 18 dah, larvae fed CY and CA treatments were largest in size, with larvae fed RA displaying the lowest growth, with no significant differences in survival among the dietary treatments. The highest DHA contents were found in ABT larvae fed CC and CY, whereas the lowest contents were found in RA-fed larvae. Indeed, larvae fed RA showed the highest level of the intermediate product n-3 docosapentaenoate, which could reflect increased activity of the biosynthetic pathway although this was not supported by gene expression data., SI

Salinity is one of the important factors affecting the physiological state of crustaceans in marine environments. Lipid plays major roles in energy supply and is main sources of essential fatty acids for membrane integrity, which is critical in adaptations to changes in salinity. Here we evaluated the effects of salinity (medium, 23 ppt and low, 4 ppt) and dietary lipid source (fish oil, FO and soybean oil, SO) on intestinal health of the marine crustacean mud crab Scylla paramamosain. The results indicated that low salinity and dietary SO (LSO group) significantly affected intestinal histomorphology, with a significant decrease of intestinal fold height and width as well as down-regulation of intestinal mRNA levels of tight junction genes compared to crab reared at medium salinity and fed FO diets (MFO group). Crabs reared at low salinity and fed SO showed an increased inflammatory response in intestine, which stimulated a physiological detoxification response together with apoptosis compared to crab in the MFO group. Low salinity and SO diets also could be responsible for multiply the pathogenic bacteria of Photobacterium and inhibit the beneficial bacteria of Firmicutes and Rhodobacteraceae in intestine, and act on a crucial impact on the development of intestinal microbial barrier disorders. The results of microbial function predictive analysis also support these inferences. The findings of the present study demonstrated that soybean oil as the main dietary lipid source could exacerbate the adverse effects of low salinity on intestinal health of mud crab, and provided evidence suggesting that dietary lipid source and fatty acid composition may play vital roles in intestinal health and the process of adaptation to environmental salinity in marine crustaceans.

An 8-week feeding trial was conducted to evaluate effects of dietary organic zinc (zinc amino acid chelate) on growth performance, mineral bioaccumulation in whole body, hepatopancreas and carapace, innate immune response and antioxidant capacity of juvenile Pacific white shrimp Litopenaeus vannamei. Five isonitrogenous and isolipidic diets were formulated to contain different zinc levels of 46.4 (basal diet), 65.5, 85.9, 108.4 and 130.6 mg kg−1. Dietary zinc level significantly influenced growth and feed utilization, with the lowest weight gain and highest feed conversion ratio observed in shrimp fed the basal diet. The optimal dietary zinc requirement was estimated to be 104.8 mg kg-1 for juvenile Pacific white shrimp. Shrimp fed the diet containing 130.6 mg kg−1 Zn had the highest zinc concentration in hepatopancreas and carapace, but there were no significant differences in calcium or phosphorus concentration in tissues. Dietary Zn increased the activities of lysozyme, alkaline phosphatase and polyphenol oxidase in hepatopancreas. Shrimp fed the diets supplemented with zinc had significantly higher activity of Cu/Zn SOD and lower content of malondialdehyde in hepatopancreas. The expression levels of toll, imd, lzm, proPO and alp involved in immunity and Cu/Zn sod related to oxidation resistance were up-regulated. Zinc also promoted the expression levels of mt and mtf-1, and up-regulated the expression of SLC39 family genes (zip3, zip9, zip11, zip14) in hepatopancreas. These data provided novel insights in the potential mechanism of organic zinc-induced enhancement of immunity and antioxidant capacity in Pacific white shrimp.

An 8-week feeding trial was conducted to determine the optimal dietary docosahexaenoic acid/eisosapentaenoic acid (DHA/EPA) ratio of mud crab (Scylla paramamosain) supplied with optimal n-3 LC-PUFA at two dietary lipid levels. Eight isonitrogenous diets were formulated to contain 7% and 12% crude lipid, each with DHA/EPA ratios of 0.6, 1.2, 2.3 and 3.2, respectively. Each diet was randomly assigned to triplicate groups of 30 juvenile mud crabs (initial weight 20.9 ± 0.6 g) that were stocked in single crab cells. In crabs fed 7% lipid, the diet with a DHA/EPA ratio of 2.3 showed significantly higher weight gain than crabs fed the other ratios while in crabs fed 12% lipid, lower weight gain and specific growth rate were observed in crabs fed the diet with a DHA/EPA ratio of 0.6 than crabs fed the other ratios. Lipid content in hepatopancreas significantly increased as dietary DHA/EPA ratio increased from 1.2 to 2.3 in crabs fed 7% lipid, while no differences were observed among crabs fed the diets with DHA/EPA ratios higher than 0.6 when fed 12% lipid. Total fatty acid and DHA contents and DHA/EPA ratio showed increasing, and EPA decreasing, trends in muscle and hepatopancreas with increased dietary DHA/EPA ratio, at both dietary lipid levels. The hemolymph triacylglycerol and total cholesterol contents were higher in crabs fed dietary DHA/PA ratios of 1.2 and 2.3 than those fed ratios of 0.6 and 3.2 at 7% dietary lipid, and lowest low and high-density lipoprotein cholesterol contents were observed in crabs fed DHA/EPA dietary ratios of 0.6 and 3.2 at 7% and 12% lipid, respectively. The expression levels of fas, aco3 and fatp4 were significantly up-regulated, and cptI, hsl and ldlr were down-regulated, with increased dietary DHA/EPA ratio in crabs fed 7% lipid. In crabs fed 12% lipid, the expression levels of g6pd, 6pgd, srebp-1, aco1 and fatp4 were down-regulated, and fabp-1 was up-regulated, with increased dietary DHA/EPA ratio. The expression levels of elovl4 and Δ6 fad initially increased and then decreased as dietary DHA/EPA ratio increased from 0.6 to 3.2 in crabs fed both 7% and 12% lipid. Based on analysis of weight gain versus dietary DHA/EPA ratio, the optimal dietary DHA/EPA ratios of mud crab S. paramamosa were estimated to be 2.2 and 1.2 when supplied with optimal n-3 LC-PUFA at 7% and 12% lipid, respectively.

Aquaculture, the fastest growing food production sector cannot continue to rely on finite stocks of marine fish as the primary source of the omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA), eicosapentaenoic acid (EPA; 20:5n3) and docosahexaenoic acid (DHA; 22:6n-3), for feeds. A four-month feeding trial was conducted to investigate the impact of a de novo oil, with high levels of EPA and DHA, obtained from transgenic Camelina sativa on growth performance, tissue fatty acid profiles, and expression of lipid metabolism genes when used as a replacement for fish oil in feed for European seabass (Dicentrachus labrax). Triplicate groups of 50 juvenile fish (initial weight 16.7 ± 0.92 g) per tank were fed for 4 months with one of three isolipidic and isoproteic experimental diets consisting of a standard diet containing a commercial blend of fish oil and rapeseed oil (CFO), a diet containing transgenic Camelina oil (TCO), or a blend of fish oil and rapeseed oil with enhanced levels of EPA and DHA (EFO) formulated to match the n-3 LC-PUFA profile of the TCO feed. Final weight of fish fed the GM-derived oil was not different to fish fed either CFO or EFO. Slight lower growth performance of fish fed TCO at the beginning of the trial was related to transient reduced feed intake, possibly caused by glucosinolates in the raw Camelina sativa oil. The GM-derived oil improved the nutritional quality of the fish fillet by enhancing total n-3 PUFA levels compared to the fish fed the other two feeds, and maintained flesh EPA and DHA at the same levels as in fish fed the diets containing fish oil. The metabolic response in liver and intestine was generally relatively mild although diets TCO and EFO seemed to trigger a metabolic response consisting of an up-regulation of both β-oxidation (cpt1a) and fatty acid transport (fabp1), possibly reflecting higher levels of LC-PUFA. Overall, the present study indicated that an oil of terrestrial origin, Camelina sativa, when engineered to contain high levels of EPA and DHA can replace fish oil in feeds for European seabass with no detrimental impact on growth or feed efficiency, while also maintaining or increasing tissue n-3 LC-PUFA contents.

Reducing the dependency of fishfeed for marine ingredients and species diversification are both considered crucial factors for the sustainable development of aquaculture. The substitution of fish oil (FO) by vegetable oils (VO) in aquafeeds is an economically feasible solution. However, such substitution may compromise the fish flesh content of essential n-3 long chain polyunsaturated fatty acids (n-3 LC-PUFA) and, therefore, its nutritional value for human consumption. Likewise, there is a wide range of strategies to select new target species for sector diversification, among which, the capacity to biosynthesize n-3 LC-PUFA from their C18 precursors abundant in VO might be considered as a fair preliminary strategy. Therefore, the aim of the present study was to analyze the metabolic fate of [1-14C] labeled 18:2n-6, 18:3n-3, 20:5n-3 and 22:6n-3 in isolated hepatocytes and enterocytes from wild individuals of three fish species with different trophic level: the marine herbivorous salema (Sarpa salpa), the strict carnivorous sand sole (Pegusa lascaris) and the omnivorous thicklip grey mullet (Chelon labrosus). These species were selected for their phylogenetic proximity to consolidated farmed species such as gilthead seabream (Sparus aurata), senegalese sole (Solea senegalensis), and golden grey mullet (Liza aurata), respectively. The study also assessed the molecular cloning, functional characterization and tissue distribution of the fatty acyl elongase (Elovl) gene, elovl5, involved in the biosynthetic metabolism of n-3 LC-PUFA. The three species were able to biosynthesize docosahexaenoic acid (22:6n-3). S. salpa seems to have similar biosynthetic capacity than S. aurata, with a fatty acyl desaturase 2 (Fads2), with Δ6, Δ8 and Δ5 activities. P. lascaris showed a wider Fads2 activity repertory than S. senegalensis, including Δ4 and residual Δ6/Δ5 activities. In C. labrosus, both Δ8 and Δ5 activities but not the Δ6 described for L. aurata were detected in the incubated cells. Elongation from C18 and C20 precursors to C20 and C22 products occurred in hepatocytes and enterocytes as well as in the functional characterization of Elovl5 by heterologous expression in yeast. Elovl5 showed a species specific expression pattern, with the highest rates observed in the liver, gut and brain in S. salpa and P. lascaris, and in the brain for C. labrosus. In summary, the LC-PUFA biosynthesis capacity from S. salpa, P. lascaris and C. labrosus greatly resembled that of their phylogenetic closer species. The three studied species could be further explored as candidates for the aquaculture diversification from their potential ability to biosynthesize LC-PUFA., This study was funded by Ministerio de Economía y Competitividad (AGL2015-70994-R). A. Galindo and M. Marrero are supported by a PhD grant by Cajasiete and Gobierno de Canarias, respectively. Dr. Covadonga Rodríguez and Dr. Ana Bolaños are members of the Instituto de Tecnologías Biomédicas de Canarias (ITB).

The increase in the global popularity and production of farmed Atlantic salmon (Salmo salar L.) has led to compositional changes in their feeds that can potentially diminish their nutritive value. Thus, the aim of the study was to compare the lipid, protein, fatty acid (omega-3) and mineral contents of salmon fillet portions available in the UK and estimate their contribution towards consumer dietary intake levels. Twenty pre-packaged fresh salmon fillets, encompassing all ranges (value, standard, premium and organic) and farmed origins (Scotland and Norway) were purchased from 10 main UK-wide retailers and analysed for their nutritional compositions. Lipid contents were between 11.2–16.3 % wet weight (ww), except the Retailer 10 value product which was significantly lower due to a high proportion of tail pieces. No difference in protein contents (17.5–20.2 % ww) were observed between fillets. However, fatty acid profiles showed marked variations between samples with marker fatty acids 18:1n-9 (24.3–42.0 %), 18:2n-6 (8.3–15.1 %) and 18:3n-3 (2.6–8.1 %) reflecting the differing levels of vegetable oil inclusion and eicosapentaenoic and docosahexaenoic acids (EPA + DHA, 5.6–16.6 %) indicating the level of marine oils included within salmon feeds. Consequently, EPA + DHA contents varied from 0.88 to 2.36 g EPA + DHA.130 g−1 flesh ww, equivalent to supplying 26–67 % of the recommended 3.5 g EPA + DHA weekly intake suggested for optimal cardiac health in adults. Similarly, selenium contents differed significantly between samples delivering between 13.9–55.5 % and 17.3–69.3 % of the 75 and 60 μg day−1 UK intake for males and females, respectively. Additionally, EPA + DHA and selenium contents were both affected by farmed origin, reflecting differences in production strategies of the two salmon producing nations. Overall, the study highlights the contrasting nutritional profiles of farmed salmon fillets available to consumers based on retailer requirements (healthy versus sustainable product) and how this can affect the recommended dietary intakes from a human nutrition perspective.

Skull, pectoral and pelvic fin bone structures in longfin yellowtail Seriola rivoliana were studied from 3.43 ± 0.15 to 16.20 ± 0.73 mm standard length (SL) specimens. The S. rivoliana skull started to mineralize with the appearance of the parasphenoid and maxillary by 3.43 ± 0.15 mm SL at the neurocranium and jaw regions, respectively. The first pectoral structure to mineralize was the cleithrum at 3.75 ± 0.14 mm SL shortly followed by the supracleithrum and posttemporal. The pelvic fin started by 6.16 ± 0.32 mm SL with the spine and continued with the soft rays and basipterygium. The present study determined the onset of the skull, pectoral and pelvic fin mineralization. These results might be used as a reference for future studies in S. rivoliana or related species.

Fish differ in their capacity for endogenous synthesis of long-chain (C20–24) polyunsaturated fatty acids (LC-PUFA) from dietary C18 precursors (α-linolenic and linoleic acids). Understanding this capacity is of benefit to fish feed formulation. This, together with the importance of fish as the primary source of omega-3 LC-PUFA in the human diet has necessitated the rigorous study of the biochemical and molecular mechanisms involved in the LC-PUFA biosynthesis pathway in fish species. Studies have shown the potential of a species for LC-PUFA biosynthesis is associated with the complement and function of fatty acyl desaturase (fads) and elongase of very long chain fatty acid (elovl) gene it possesses. The present study therefore aimed to investigate these genes in the African catfish (Clarias gariepinus), the most commercially important farmed fish species in Sub-Saharan Africa. A fads2 and an elovl2 cDNA were cloned containing open reading frames (ORF) of 1338 base pair (bp) and 864 bp specifying proteins of 445 and 287 amino acids, respectively. Functional characterisation by heterologous expression in yeast showed that the Fads2 was bifunctional with Δ5Δ6 activities catalysing the desaturation of both 18:3n-3 and 20:4n-3 and their corresponding n-6 fatty acids, 18:2n-6 and 20:3n-6. The Elovl2 showed activity towards C18, C20 and C22 PUFA with highest activity towards C20 and C22 PUFA. Tissue expression analysis showed a typical freshwater species expression pattern; higher expression in the liver compared to brain and all other tissues with the exception of elovl5 which showed highest expression in the intestine. Consistent with feeding studies of typical freshwater fish species that show their essential fatty acid requirement can be satisfied by dietary C18 PUFA, the present study confirms that the LC-PUFA biosynthesis pathway is active in the African catfish C. gariepinus. Statement of relevance Clarias gariepinus has enzymes to use dietary vegetable oils.

Atlantic salmon (Salmo salar) were fed 6 experimental diets containing three levels of arachidonic acid (ARA) (0.18 g kg−1, 0.28 g kg−1 and 0.63 g kg−1 for low, medium and high levels, respectively) and two levels of vitamin E (150 and 730 mg kg−1 for low and high levels, respectively). At the end of the experimental period, fatty acids in the liver and immunity markers (lysozyme activity, respiratory burst and phagocytic activity) were determined and fish subjected to a challenge test against the salmonid rickettsial syndrome (SRS) pathogen. ARA, vitamin E or their interaction did not exert an effect on fish performance, whereas ARA alone clearly increased the deposition of ARA. Dietary vitamin E only enhanced liver vitamin E deposition, while the interaction of ARA and vitamin E influenced lysozyme activity and EPA/ARA ratio pointing out the effect of both nutrients on the fish immune system and metabolism. Only the medium concentration contributed to reducing mortality when the fish were exposed to the SRS pathogen. In conclusion, different levels of supplementation with ARA and vitamin E in the diet had no effect on productivity, but did have effects on immune markers and cumulative mortality when fish were exposed to the SRS pathogen.

Increased substitution of marine ingredients by terrestrial plant products in aquafeeds has been proven to be suitable for Atlantic salmon farming. However, a reduction inn-3 long-chain PUFA is a consequence of this substitution. In contrast, relatively little attention has been paid to the effects of fishmeal and oil substitution on levels of micronutrients such as Se, considering fish are major sources of this mineral for human consumers. To evaluate the effects of dietary marine ingredient substitution on tissue Se distribution and the expression of Se metabolism and antioxidant enzyme genes, Atlantic salmons were fed three feeds based on commercial formulations with increasing levels of plant proteins (PP) and vegetable oil. Lipid content in flesh did not vary at any sampling point, but it was higher in the liver of 1 kg of fish fed higher PP. Fatty acid content reflected dietary input and was related to oxidation levels (thiobarbituric acid-reactive substances). Liver had the highest Se levels, followed by head kidney, whereas the lowest contents were found in brain and gill. The Se concentration of flesh decreased considerably with high levels of substitution, reducing the added value of fish consumption. Only the brain showed significant differences in glutathione peroxidase, transfer RNA selenocysteine 1-associated protein 1b and superoxide dismutase expression, whereas no significant regulation of Se-related genes was found in liver. Although Se levels in the diets satisfied the essential requirements of salmon, high PP levels led to a reduction in the supply of this essential micronutrient.

Copper is a widespread pollutant in marine environments, and marine animals can ingest large amounts of copper through the food chain. Here, an 8-week feeding trial was designed to investigate the effects of different dietary copper levels on coloration, copper bioaccumulation, stress response and oxidation resistance of juvenile mud crab Scylla paramamosain. The results indicated that crabs fed the diet with 162 mg/kg copper exhibited a dark-blue carapace and hemolymph. The accumulation of copper in tissues was positively correlated with the level of copper in feed. High/excess dietary copper (162 mg/kg) up-regulated the expression of stress response related genes, and reduced the expression/activities of anti-oxidation genes/enzymes. The activity of phenoloxidase decreased significantly when dietary copper level was 86-162 mg/kg, and the expression of hemocyanin was up-regulated in crab fed the diets with 28-162 mg/kg copper. Overall, the results of the present study indicated that high dietary copper led to parachrea in carapace and hemolymph of mud crab, and caused copper deposition abnormality in carapace and hepatopancreas. The data suggested that the toxic effects of dietary copper were concentration-dependent such that, excess dietary copper (162 mg/kg) had adverse impacts on oxidation resistance.

A common-garden experiment was carried out to compare two genetically distinct strains of Atlantic salmon (Salmo salar) fed diets with either high (CHO) or low (NoCHO) digestible carbohydrate (starch). Twenty salmon from either a commercial farmed strain (F) or a land-locked population (G) were placed in two tanks (10 fish of each population in each tank) and fed either CHO or NoCHO feeds. At the end of the experiment fish were fasted for 8 h, euthanized and blood and liver collected. Both diet and population had an effect on circulating glucose levels with G showing hypoglycaemia and dietary starch increasing this parameter. In contrast, G showed increased plasma triacylglycerol levels regardless of dietary treatment suggesting faster conversion of glucose to triacylglycerol. This different ability to metabolize dietary starch among strains was also reflected at a molecular (gene) level as most of the metabolic pathways evaluated were mainly affected by the factor population rather than by diet. The data are promising and suggest different regulatory capacities toward starch utilization between land-locked salmon and the farmed stock. Further analyses are necessary in order to fully characterize the capacity of land-locked salmon to utilize dietary carbohydrate. © 2018 Betancor, Olsen, Marandel, Skulstad, Madaro, Tocher and Panserat. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Facing a bottleneck in the growth of aquaculture, and a gap in the supply and demand of the highly beneficial n-3 long-chain PUFA (LC-PUFA), sustainable alternatives to traditional marine-based feeds are required. Therefore, in the present trial, a novel oil obtained from a genetically engineered oilseed crop, Camelina sativa, that supplied over 25 % n-3 LC-PUFA was tested as a sole dietary-added lipid source in Atlantic salmon (Salmo salar) feed. Three groups of fish were fed three experimental diets for 12 weeks with the same basal composition and containing 20 % added oil supplied by either a blend of fish oil and rapeseed oil (1:3) (COM) reflecting current commercial formulations, wild-type Camelina oil (WCO) or the novel transgenic Camelina oil (TCO). There were no negative effects on the growth, survival rate or health of the fish. The whole fish and flesh n-3 LC-PUFA levels were highest in fish fed TCO, with levels more than 2-fold higher compared with those of fish fed the COM and WCO diets, respectively. Diet TCO had no negative impacts on the evaluated immune and physiological parameters of head kidney monocytes. The transcriptomic responses of liver and mid-intestine showed only mild effects on metabolism genes. Overall, the results clearly indicated that the oil from transgenic Camelina was highly efficient in supplying n-3 LC-PUFA providing levels double that obtained with a current commercial standard, and similar to those a decade ago before substantial dietary fishmeal and oil replacement. This article has been accepted for publication in British Journal of Nutrition . This version is free to view and download for private research and study only. Not for re-distribution, re-sale or use in derivative works. © The Authors 2018

The development of formulated diets and feeds is essential to increase production of farmed tuna species. There is limited knowledge of this topic, mainly on Pacific Bluefin tuna (Thunnus orientalis) in Japan, whereas no major attempts have been made with Atlantic Bluefin tuna (Thunnus thynnus; ABT). In the present study, two trials were performed using inert formulated diets as on-growing feeds for weaned ABT juvenile in order to establish adequate dietary levels of both lipid and omega-3 long-chain polyunsaturated fatty acids (LC-PUFA). In a first trial, ABT (initial weight = 2.9±0.9g) were fed for 10 days with either a commercial (Magokoro®, MGK) or two experimental feeds with two different lipid levels (15 or 20%) using krill oil (KO) as the single lipid source in order to estimate the suitable lipid content. Fish fed MGK displayed the highest growth, followed by 15KO, with no differences in fish survival. Thus, a lipid content of 15% was considered better than 20% for ABT juveniles. In the second trial, fish (initial weight = 3.3 ± 0.6g) were fed either MGK, 15KO or a feed containing 15% lipid with a combination (1:1, v/v) KO and rapeseed oil (RO) (15KORO). Fish fed 15KO and 15KORO showed the highest growth in terms of weight and fork length (including weight gain and SGR). Increasing dietary lipid level or adding RO to the feeds did not increase liver lipid content. The liver fatty acid profile largely reflected dietary intake confirming very limited LC-PUFA biosynthetic activity for this teleost species. In this respect, liver of fish fed 15KO and 20KO displayed the highest contents of docosahexaenoic acid (DHA). The hepatic expression of genes of lipid and fatty acid metabolism, transcription factors, and antioxidant enzymes was investigated with many of the genes showing regulation by both dietary lipid and LC-PUFA contents. The present study showed promising results that suggested ABT juveniles can be on grown on inert dry feeds that supported good fish growth and the accumulation of the health-promoting fatty acid DHA. Further studies are required in order to fully elucidate lipid and fatty acid requirements of this iconic species regarding dietary sources and production costs., SI

The present study aimed to evaluate the effect of the supplementation of different crab zoeas to enriched Artemia basal diet for Octopus vulgaris paralarvae during the first month of life. Paralarvae were fed using enriched Artemia nauplii alone and Artemia co-fed either first zoea stages of Grapsus adscensionis or Plagusia depressa. The experiment was carried out over a period of 28 days, in 0.12 m3 tanks with a flow-through rearing system. Growth in dry weight as well as mantle length and width were assessed weekly. Additionally, prey and paralarvae fatty acid composition and digestive gland (DG) histology were evaluated. Addition of low amounts of crab zoeas (approx. 100 indv. L−1 day−1) provided during critical life stages of O. vulgaris proved to be good enough to improve paralarvae growth and survival in comparison with those fed exclusively on enriched Artemia. These results were supported by the finding of a higher number of glycoprotein absorption vacuoles in the DG from paralarvae co-fed crab zoeas, suggesting a higher feeding activity. In addition, fatty acid analysis of crab zoea showed that these are good sources of dietary arachidonic and eicosapentaenoic acids during the octopus planktonic life stage, whereas the low docosahexaenoic (DHA) content suggests the use of additional DHA sources or higher zoea densities to meet paralarvae nutritional demand to carry out a successful metamorphosis to benthic life.

In vertebrates, selenium (Se) is an essential micronutrient for vertebrates that is involved in antioxidant protection and thyroid hormone regulation among other roles and functions through its incorporation into proteins, the selenoproteins. Long-chain polyunsaturated fatty acids, such as docosahexaenoic acid (DHA), are essential nutrients for fish although high dietary levels may lead to increased oxidative stress due to the high degree of unsaturation. The present study investigated the effects of Se supplementation on zebrafish, Danio rerio, oxidative status together with selenoprotein expression profiles when subjected to a high-DHA diet. Fish were fed for 8 weeks with one of the four experimental diets, containing high or low DHA in combination with or without organic Se (7 mg/kg). Fish performance, Se content, fatty acid composition and TBARS of zebrafish were determined, as well as gene expression of selected selenoproteins in liver and muscle. The Se levels in whole fish reflected dietary content. High dietary DHA increased oxidative stress as indicated by reduced growth and high TBARS content, although Se supplementation reduced oxidation. The expression patterns of selenoproteins varied between liver and muscle with only deiodinase type II displaying a transcriptional response when high dietary Se was supplied. High dietary DHA decreased selenoprotein W expression in muscle and sps2 expression in liver regardless of the dietary Se content. These data suggest that oxidative stress protection associated with a high dietary intake of Se may not be solely mediated by transcriptional changes in teleost selenoprotein expression.