More Than an Antioxidant: Role of Dietary Astaxanthin on Lipid and Glucose Metabolism in the Liver of Rainbow Trout (Oncorhynchus mykiss).

This study investigated the influence of dietary astaxanthin (AX) on glucose and lipid metabolism in rainbow trout liver. Two iso-nitrogenous and iso-lipidic diets were tested for 12 weeks in rainbow trout with an initial mean weight of 309 g. The S-ASTA diet was supplemented with 100 mg of synthetic AX per kg of feed, whereas the control diet (CTRL) had no AX. Fish fed the S-ASTA diet displayed lower neutral and higher polar lipids in the liver, associated with smaller hepatocytes and lower cytoplasm vacuolization. Dietary AX upregulated adipose triglyceride lipase (atgl), hormone-sensitive lipase (hsl2) and 1,2-diacylglycerol choline phosphotransferase (chpt), and downregulated diacylglycerol acyltransferase (dgat2), suggesting the AX's role in triacylglycerol (TAG) turnover and phospholipid (PL) synthesis. Dietary AX may also affect beta-oxidation with the upregulation of carnitine palmitoyltransferase 1 (cpt1α2). Although hepatic cholesterol levels were not affected, dietary AX increased gene expression of sterol regulatory element-binding protein 2 (srebp2). Dietary AX upregulated the expression of 6-phosphogluconate dehydrogenase (6pgdh) and downregulated pyruvate kinase (pkl). Overall, results suggest that dietary AX modulates the oxidative phase of the pentose phosphate pathway and the last step of glycolysis, affecting TAG turnover, ß-oxidation, PL and cholesterol synthesis in rainbow trout liver.

Stable isotope and fatty acid analysis reveal the ability of sea cucumbers to use fish farm waste in integrated multi-trophic aquaculture.

Stable isotope ratios, carbon (δ13C) and nitrogen (δ15N), and fatty acids validated the trophic connection between farmed fish in a commercial nearshore fish farm and sea cucumbers in the Mediterranean Sea. This dual tracer approach evaluated organic matter transfer in integrated multi-trophic aquaculture (IMTA) and the ability of sea cucumbers to incorporate fish farm waste (fish faeces and uneaten artificial fish feed) into their tissue. Between October 2018 and September 2019, Holothuria (Roweothuria) poli Delle Chiaje, 1824, co-cultured at IMTA sites directly below one of the commercial fish cage , at 10 m and 25 m from the selected fish cage, and at two reference sites over 800 m from the fish farm. Sea cucumbers were sampled from each site in February, May and September, except at 0 m due to mass mortalities recorded here in the first month of study. Isotopic mixing models revealed that fish farm organic waste was the dominant dietary source for H. poli in IMTA at 10 m and 25 m from the cage. The contribution of marine plant-derived organic matter, Posidonia oceanica leaves and rhizomes, was least important. The isotopic signatures of sea cucumber tissues at reference sites were not explained by the sampled food resources. Importantly, fatty acid profiling revealed a high abundance of individual terrestrial plant fatty acids, such as oleic (18:1n-9), linoleic (18:2n-6) and eicosenoic (20:1n-9) acids in sea cucumber tissue at 10 m and 25 m from the fish cage, presumably linked to the terrestrial plant oil content of the fish feeds. At the reference sites, sea cucumber tissues were characterised by higher relative abundance of arachidonic acid (20:4n-6) acid, and the natural marine-based eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids. These analyses revealed important differences in the composition of H. poli between the IMTA and reference locations, driven by aquaculture-derived waste near fish cages. Moreover, this study revealed temporal variation in food availability and quality, and possible differences in the physiological responses of H. poli. Stable isotope analysis and fatty acid profiling provided complementary evidence for the important dietary preferences of H. poli and validated the potential of sea cucumbers to uptake aquaculture organic waste as part of inshore fish-sea cucumber IMTA. It reveals the important implications that an established trophic link has on the viability of using sea cucumbers for the development of IMTA and the sustainable expansion of aquaculture.

Molecular Antioxidant Functions are Enhanced in Atlantic Bluefin Tuna (Thunnus Thynnus, L.) Larvae Fed Selenium-Enriched Rotifers Brachionus Rotundiformis.

Selenium (Se) is an essential trace element for fish with more than 40 selenoproteins identified, many exhibiting antioxidant functions. This study investigated the effect of dietary Se supplementation on physiological parameters, selenoprotein and antioxidant enzyme gene expression in Atlantic bluefin tuna (ABT, Thunnus thynnus) larvae. First-feeding ABT larvae were divided into triplicate groups and fed rotifers Brachionus rotundiformis enriched with five different levels of Se (0, 3, 10, 30, and 100 µg Se·L-1) until 14 days after hatching. Both rotifers and ABT larvae effectively accumulated Se achieving maximum levels in the Se100 treatment (30.05 µg Se·g-1 and 194 ± 38 µg Se·g-1 dry mass, respectively). Larvae showed highest total length when fed Se3 rotifers, whereas flexion index was highest in larvae fed Se10. Selenium supplementation increased the gene expression of selenoproteins gpx1, msrb1, trxr2, selenom, selenop, and selenoe compared to the non-supplemented control (Se0), but only marginal differences were detected between supplementation levels. In contrast, expression of the antioxidant enzymes cat and sod1 were lowest in larvae fed Se100. To conclude, non-Se-enriched rotifers may be suboptimal for first feeding ABT larvae, which showed improved selenoprotein and antioxidant gene expression when fed a diet containing 4.42 µg Se·g-1 dry mass.

Oil from transgenic Camelina sativa containing over 25 % n-3 long-chain PUFA as the major lipid source in feed for Atlantic salmon (Salmo salar).

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.

The potential impact of compositional changes in farmed fish on its health-giving properties: is it time to reconsider current dietary recommendations?

Assessment of national dietary guidelines in a number of European countries reveals that some are based on cohort studies, focusing on total seafood consumption, while others are based on the content of EPA and DHA, distinguishing between oily and other fish. The mean actual intake of fish in most countries is around or below the recommended intake, with differences in intake of fish being present between sex and age groups. Many people do not reach the national recommendation for total fish intake. Dietary recommendations for fish and EPA/DHA are based mainly on data collected more than 10 years ago. However, methods of farmed fish production have changed considerably since then. The actual content of EPA and DHA in farmed salmon has nearly halved as the traditional finite marine ingredients fish meal and fish oil in salmon diets have been replaced with sustainable alternatives of terrestrial origin. As farmed salmon is an important source of EPA and DHA in many Western countries, our intake of these fatty acids is likely to have decreased. In addition, levels of vitamin D and Se are also found to have declined in farmed fish in the past decade. Significant changes in the EPA and DHA, vitamin D and Se content of farmed fish means that average intakes of these nutrients in Western populations are probably lower than before. This may have consequences for the health-giving properties of fish as well as future dietary recommendations for fish intake.

An oil containing EPA and DHA from transgenic Camelina sativa to replace marine fish oil in feeds for Atlantic salmon (Salmo salar L.): Effects on intestinal transcriptome, histology, tissue fatty acid profiles and plasma biochemistry.

New de novo sources of omega 3 (n-3) long chain polyunsaturated fatty acids (LC-PUFA) are required as alternatives to fish oil in aquafeeds in order to maintain adequate levels of the beneficial fatty acids, eicosapentaenoic and docosahexaenoic (EPA and DHA, respectively). The present study investigated the use of an EPA+DHA oil derived from transgenic Camelina sativa in Atlantic salmon (Salmo salar) feeds containing low levels of fishmeal (35%) and fish oil (10%), reflecting current commercial formulations, to determine the impacts on tissue fatty acid profile, intestinal transcriptome, and health of farmed salmon. Post-smolt Atlantic salmon were fed for 12-weeks with one of three experimental diets containing either a blend of fish oil/rapeseed oil (FO), wild-type camelina oil (WCO) or transgenic camelina oil (DCO) as added lipid source. The DCO diet did not affect any of the fish performance or health parameters studied. Analyses of the mid and hindgut transcriptomes showed only mild effects on metabolism. Flesh of fish fed the DCO diet accumulated almost double the amount of n-3 LC-PUFA than fish fed the FO or WCO diets, indicating that these oils from transgenic oilseeds offer the opportunity to increase the n-3 LC-PUFA in farmed fish to levels comparable to those found a decade ago.

Nutritional Evaluation of an EPA-DHA Oil from Transgenic Camelina sativa in Feeds for Post-Smolt Atlantic Salmon (Salmo salar L.).

Vegetable oils (VO) are possible substitutes for fish oil in aquafeeds but their use is limited by their lack of omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA). However, oilseed crops can be modified to produce n-3 LC-PUFA such as eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, representing a potential option to fill the gap between supply and demand of these important nutrients. Camelina sativa was metabolically engineered to produce a seed oil with around 15% total n-3 LC-PUFA to potentially substitute for fish oil in salmon feeds. Post-smolt Atlantic salmon (Salmo salar) were fed for 11-weeks with one of three experimental diets containing either fish oil (FO), wild-type Camelina oil (WCO) or transgenic Camelina oil (DCO) as added lipid source to evaluate fish performance, nutrient digestibility, tissue n-3 LC-PUFA, and metabolic impact determined by liver transcriptome analysis. The DCO diet did not affect any of the performance or health parameters studied and enhanced apparent digestibility of EPA and DHA compared to the WCO diet. The level of total n-3 LC-PUFA was higher in all the tissues of DCO-fed fish than in WCO-fed fish with levels in liver similar to those in fish fed FO. Endogenous LC-PUFA biosynthetic activity was observed in fish fed both the Camelina oil diets as indicated by the liver transcriptome and levels of intermediate metabolites such as docosapentaenoic acid, with data suggesting that the dietary combination of EPA and DHA inhibited desaturation and elongation activities. Expression of genes involved in phospholipid and triacylglycerol metabolism followed a similar pattern in fish fed DCO and WCO despite the difference in n-3 LC-PUFA contents.

Effects of decontaminated fish oil or a fish and vegetable oil blend on persistent organic pollutant and fatty acid compositions in diet and flesh of Atlantic salmon ( Salmo salar).

The health benefits of seafood are well documented and based on the unique supply of n-3 highly unsaturated fatty acids (HUFA). Aquaculture now contributes about 50 % of food-grade seafood globally and Atlantic salmon (Salmo salar) is a rich source of n-3 HUFA. However, salmon and other oily fish can accumulate lipophilic persistent organic pollutants (POP), including dioxins (PCDD/F), polychlorinated biphenyls (PCB) and polybrominated diphenyl ethers (PBDE), derived largely from feed. In the present study, triplicate groups of salmon, of initial weight 0.78 kg, were fed one of three experimental diets for 11 weeks. The diets were coated with either a northern fish oil (FO) with a high POP content (cNFO), the same oil that had been decontaminated (deNFO) or a blend of southern fish oil, rapeseed and soyabean oils (SFO/RO/SO). Dietary PCDD/F+dioxin-like PCB (DL-PCB) concentrations were 17.36, 0.45 and 0.53 ng toxic equivalents (TEQ)/kg, respectively. After 11 weeks, the flesh concentrations in fish fed the cNFO, deNFO and SFO/RO/SO diets were 6.42, 0.34 and 0.41 ng TEQ/kg, respectively. There were no differences in flesh EPA and DHA between fish fed the cNFO or deNFO diets although EPA and DHA were reduced by 50 and 30 %, respectively, in fish fed the SFO/RO/SO diet. Thus, decontaminated FO can be used to produce salmon high in n-3 HUFA and low in POP. Salmon produced using deNFO would be of high nutritional value and very low in POP and would utilise valuable fish oils that would otherwise be destroyed due to their high pollutant concentrations.