Profiling Phospholipids within Atlantic Salmon Salmo salar with Regards to a Novel Terrestrial Omega-3 Oil Source

The development and inclusion of novel oils derived from genetically modified (GM) oilseeds into aquafeeds, to supplement and supplant current terrestrial oilseeds, as well as fish oils, warrants a more thorough investigation into lipid biochemical alterations within finfish species, such as Atlantic salmon. Five tissues were examined across two harvesting timepoints to establish whether lipid isomeric alterations could be detected between a standard commercial diet versus a diet that incorporated the long-chain polyunsaturated fatty acids (LC-PUFA), EPA (eicosapentaenoic acid), and DHA (docosahexaenoic acid), derived from the GM oilseed Camelina sativa. Tissue-dependent trends were detected, indicating that certain organs, such as the brain, have a basal limit to LC-PUFA incorporation, though enrichment of these fatty acids is possible. Lipid acyl alterations, as well as putative stereospecific numbering (sn) isomer alterations, were also detected, providing evidence that GM oils may modify lipid structure, with lipids of interest providing a set of targeted markers by which lipid alterations can be monitored across various novel diets

Comparative Fatty Acid Composition of Eggs from White Bass Fed Live Food or Commercial Feed

We evaluated the influence of two broodstock feeding practices on fatty acid composition and viability of eggs in white bass Morone chrysops. The two dietary groups tested were (1) white bass females fed a commercially formulated feed (crude protein, 45%; crude fat, 16%) and (2) white bass females maintained on live food (fathead minnow Pimephales promelas and golden shiners Notemigonus crysoleucas). Significant differences existed between the dietary treatments in egg fatty acid levels. Eggs of white bass fed live food contained more 11-octadecenoic acid (18:1[n-7]), α-linolenic acid (18:3[n-3]), arachidonic acid (20:4[n-6]), docosahexaenoic acid (22:6[n-3]), and total n-3 fatty acids than eggs of fish fed formulated feed. Conversely, eggs of fish fed the formulated feed contained more oleic acid (18:1[n-9]), linoleic acid (18:2[n-6]), and total monoeic acids. Female white bass fed live food produced significantly more viable eggs (68.0 ± 2.0%) at 48 ± 2 h posthatch than did females fed the commercial feed (57.0 ± 2.0% [mean ± SD]). We found that the egg fatty acids of white bass are significantly affected by the diet of the female and suggest that the fatty acid composition of eggs contribute to overall reproductive success and viability of progeny. More information on the nutrient requirements of piscivorous broodstock is needed to allow feeds to be formulated to enhance the viability of eggs and fry

Polyunsaturated fatty acid metabolism in Atlantic salmon (Salmo salar) undergoing parr-smolt transformation and the effects of dietary linseed and rapeseed oils

Duplicate groups of Atlantic salmon parr were fed diets containing either fish oil (FO), rapeseed oil (RO), linseed oil (LO) or linseed oil supplemented with arachidonic acid (20:4n-6; AA) (LOA) from October (week 0) to seawater transfer in March (week 19). From March to July (weeks 20-34) all fish were fed a fish oil-containing diet. Fatty acyl desaturation and elongation activity in isolated hepatocytes incubated with [1-14C]18:3n-3 increased in all dietary groups, peaking in early March about one month prior to seawater transfer. Desaturation activities at their peak were significantly greater in fish fed the vegetable oils, particularly RO, compared to fish fed FO. Docosahexaenoic acid (22:6n-3:DHA) and AA in liver and gill polar lipids (PL) increased in all dietary groups during the freshwater phase whereas eicosapentaenoic acid (20:5n-3;EPA) increased greatly in all groups after seawater transfer. The AA/EPA ratio in tissue PL increased up to seawater transfer and then decreased after transfer. AA levels and the AA/EPA ratio in gill PL were generally higher in the LOA group. The levels of 18:3n-3 in muscle total lipid were increased significantly in the LO, LOA and, to a lesser extent, RO groups prior to transfer but were reduced to initial levels by the termination of the experiment (week 34). In contrast, 18:2n-6 in muscle total lipid was significantly increased after 18 weeks in fish fed the diets supplemented with RO and LO, and was significantly greater in the FO and RO groups at the termination of the experiment. Gill PGF production showed a large peak about two months after transfer to seawater. The production of total PGF post-transfer was significantly lower in fish previously fed the LOA diet. However, plasma chloride concentrations in fish subjected to a seawater challenge at 18 weeks were all lower in fish fed the diets with vegetable oils. This effect was significant in the case of fish receiving the diet with LOA, compared to those fed the diet containing FO. The present study showed that during parr-smolt transformation in Atlantic salmon there is a pre-adaptive increase in hepatocyte fatty acyl desaturation/elongation activities that is controlled primarily by environmental factors such as photoperiod and temperature but that can also be significantly modulated by diet. Feeding salmon parr diets supplemented with rapeseed or linseed oils prevented inhibition of the desaturase activities that is induced by feeding parr diets with fish oils and thus influenced the smoltification process by altering tissue PL fatty acid compositions and eicosanoid production. These effect, in turn, had a beneficial effect on the ability of the fish to osmoregulate and thus adapt to salinity changes

Risk assessment of the use of alternative animal and plant raw material resources in aquaculture feeds

A wide range of raw materials are now used routinely in aquaculture feeds throughout the world, primarily to supply protein and energy in the form of lipid from edible oils. Protein meals and oils used can generally be divided into those of plant or animal origin and many have considerable potential to supply the required dietary nutrients required by aquaculture species. However, the use of any raw material introduces a suite of risks that need to be considered to enable the production of safe, sustainable and functional feeds to underpin this sector. A lack of understanding of some of those risks can result in failure of dietary specifications being met and/or negative nutritional elements being introduced (e.g. antinutritional factors). Importantly, it is this feed that when fed to food‐producing animals is such an important element of food safety, and as such any undesirable aspects relating to feed production can also have a negative impact on the rest of the food chain. However, there is some disparity internationally among raw materials that are used and the perceptions surrounding the risk of their use. It is the scientific assessment of these risks that is the basis of this review

Mass Production of Lemna minor and Its Amino Acid and Fatty Acid Profiles

The surface floating duckweed Lemna minor (Lemnaceae) is a potential ingredient to replace the application of fish-meal in the aqua-feed. The culture technique of the duckweed was standardized in outdoor tanks and then applied in the pond. Three consecutive experiments were conducted in tanks (1.2 × 0.35 × 0.3 m). In experiment 1, four different manures were used. In manure 1 (organic manure, OM) and manure 3 (2x OM), cattle manure, poultry droppings, and mustard oil cake (1:1:1) were used; in manure 2 (inorganic fertilizer, IF), urea, potash, triple superphosphate were used; manure 4 (2x OM+IF) was a combination of manure 2 and manure 3. In experiment 2, manure 1 (OM) and manure 2 (IF) were used, and manure 3 (OM+IF) was a combination of both manures. In experiment 3, OM and IF were selected. In pond (20 × 10 × 0.5 m), OM was applied. Fresh duckweed was seeded after 5 days of manure application. In experiments 1 and 3, total production was significantly (P 0.05) difference in production between OM and IF. In pond, relative growth rate (RGR) of duckweed ranged from 0.422 to 0.073 g/g/day and total production was 702.5 Kg/ha/month (dry weight). Protein, lipid, and ash contents were higher in duckweed cultured in OM compared to IF. The duckweed was a rich source of essential (39.20%), non-essential (53.64%), and non-proteinogenic (7.13%) amino acids. Among essential amino acids, leucine, isoleucine, and valine constituted 48.67%. Glutamic acid was 25.87% of total non-essential amino acids. Citrulline, hydroxiproline, taurine, etc. were found in the duckweed. The fatty acid composition was dominated by PUFA, 60–63% of total fatty acids, largely α-linolenic acid (LNA, 18:3n-3) at around 41 to 47% and linoleic acid (LA, 18:2n-6) at 17–18%. The nutritional value of duckweeds and their production potential in the pond conditions were evaluated. Duckweed biomass may thus be used to replace commercial fish-meal that is currently used in aquaculture

Effect of salinity on the biosynthesis of n-3 long-chain polyunsaturated fatty acids in silverside Chirostoma estor

The genus Chirostoma (silversides) belongs to the family Atherinopsidae, which contains around 150 species, most of which are marine. However, Mexican silverside (Chirostoma estor) is one of the few representatives of freshwater atherinopsids and is only found in some lakes of the Mexican Central Plateau. However, studies have shown that C. estor has improved survival, growth and development when cultured in water conditions with increased salinity. In addition, C. estor displays an unusual fatty acid composition for a freshwater fish with high docosahexaenoic acid (DHA) : eicosapentaenoic acid (EPA) ratios. Freshwater and marine fish species display very different essential fatty acid metabolism and requirements and so the present study investigated long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis to determine the capacity of C. estor for endogenous production of EPA and DHA, and the effect that salinity has on these pathways. Briefly, C. estor were maintained at three salinities (0, 5 and 15 ppt) and the metabolism of 14C-labelled 18:3n-3 determined in isolated hepatocyte and enterocyte cells. The results showed that C. estor has the capacity for endogenous biosynthesis of LC-PUFA from 18-carbon fatty acid precursors, but that the pathway was essentially only active in saline conditions with virtually no activity in cells isolated from fish grown in freshwater. The activity of the LCPUFA biosynthesis pathway was also higher in cells isolated from fish at 15 ppt compared to fish at 5 ppt, The pathway was around 5-fold higher in hepatocytes compared to enterocytes, although the majority of 18:3n-3 was converted to 18:4n-3 and 20:4n-3 in hepatocytes whereas the proportions of 18:3n-3 converted to EPA and DHA were higher in enterocytes. The data were consistent with the hypothesis that conversion of EPA to DHA could contribute, at least in part, to the generally high DHA:EPA ratios observed in the tissue lipids of C. estor

Endogenous production of n-3 long-chain PUFA from first feeding and the influence of dietary linoleic acid and the α-linolenic:linoleic ratio in Atlantic salmon (Salmo salar)

Atlantic salmon (Salmo salar) possess enzymes required for the endogenous biosynthesis of n-3 long-chain polyunsaturated fatty acids (LC-PUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from a-linolenic acid (ALA). Linoleic acid (LA) competes with ALA for LC-PUFA biosynthesis enzymes leading to the production of n-6 LC-PUFA, including arachidonic acid (ARA). We aimed to quantify the endogenous production of EPA and DHA from ALA in salmon fed from first feeding on diets that contain no EPA and DHA, and to determine the influence of dietary LA and ALA:LA ratio on LC-PUFA production. Salmon were fed from first feeding for 22 weeks with three diets formulated with linseed and sunflower oils to provide ALA:LA ratios of approximately 3:1, 1:1 and 1:3. Endogenous production of n-3 LC-PUFA was 5.9, 4.4 and 2.8 mg per g fish and that of n-6 LC-PUFA was 0.2, 0.5 and 1.4 mg per g of fish in salmon fed diets with ALA:LA ratios of 3:1, 1:1 and 1:3, respectively. The ratio of n-3:n-6 LC-PUFA production decreased from 27.4 to 2.0, and DHA:EPA ratio increased and EPA:ARA and DHA:ARA ratios decreased, as dietary ALA:LA ratio decreased. In conclusion, with a dietary ALA:LA ratio of 1, salmon fry/parr produced around 28 μg n-3 LC-PUFA per g of fish per day, with a DHA:EPA ratio of 3.4. Production of n-3 LC-PUFA exceeded that of n-6 LC-PUFA by almost 9-fold. Reducing the dietary ALA:LA ratio reduced n-3 LC-PUFA production, and EPA:ARA and DHA:ARA ratios, and increased n-6 LC-PUFA production, and DHA:EPA ratio

Heritability and mechanisms of n-3 long chain polyunsaturated fatty acid deposition in the flesh of Atlantic salmon

N-3 long chain polyunsaturated fatty acids (n-3LC-PUFA) are essential components of vertebrate membrane lipids and are crucially deficient in modern Western diets. The main human dietary source for n-3LC-PUFA is fish and seafood, particularly oily fish and over 50% of global fish production is currently supplied by aquaculture. However, increasing pressure to include vegetable oils, which are devoid of n-3LC-PUFA, in aquaculture feeds reduces the content of these crucial nutrients in farmed fish flesh. The aim of this study was to measure the heritability and infer mechanisms determining flesh n-3LC-PUFA content in Atlantic salmon. This was achieved by analysing flesh lipid parameters in 48 families of Atlantic salmon, and by measuring differences in hepatic mRNA expression in families with high and low flesh n-3LC-PUFA. The results show that flesh n-3LC-PUFA level is a highly heritable trait (h2 = 0.77±0.14) and indicate the involvement of increased lipid transport, most likely in the form of very low density lipoprotein (VLDL) from liver. This increase in lipid transport may be associated with increased activity of a transcription factor, hepatic nuclear factor 4α (HNF4α), possibly as a result of family differences in transforming growth factor β1 (Tgfβ1) signalling. This study paves the way for identification of quantitative trait loci and gene interaction networks that are associated with levels of n-3LC-PUFA in fish flesh. Such markers can be used to assist the sustainable production of Atlantic salmon and provide optimal levels of critical nutrients for human consumers

Investigation of highly unsaturated fatty acid metabolism in the Asian sea bass, Lates calcarifer

Lates calcarifer, commonly known as the Asian sea bass or barramundi, is an interesting species that has great aquaculture potential in Asia including Malaysia and also Australia. We have investigated essential fatty acid metabolism in this species, focusing on the endogenous highly unsaturated fatty acid (HUFA) synthesis pathway using both biochemical and molecular biological approaches. Fatty acyl desaturase (Fad) and elongase (Elovl) cDNAs were cloned and functional characterization identified them as ∆6 Fad and Elovl5 elongase enzymes, respectively. The ∆6 Fad was equally active towards 18:3n-3 and 18:2n-6, and Elovl5 exhibited elongation activity for C18-20 and C20-22 elongation and a trace of C22-24 activity. The tissue profile of gene expression for ∆6 fad and elovl5 genes, showed brain to have the highest expression of both genes compared to all other tissues. The results of tissue fatty acid analysis showed that the brain contained more docosahexaenoic acid (DHA, 22:6n-3) than flesh, liver and intestine. The HUFA synthesis activity in isolated hepatocytes and enterocytes using [1-14C]18:3n-3 as substrate was very low with the only desaturated product detected being 18:4n-3. These findings indicate that L. calcarifer display an essential fatty acid pattern similar to other marine fish in that they appear unable to synthesize HUFA from C18 substrates. High expression of ∆6 fad and elovl5 genes in brain may indicate a role for these enzymes in maintaining high DHA levels in neural tissues through conversion of 20:5n-3

Egg quality determinants in cod (Gadus morhua L.): egg performance and lipids in eggs from farmed and wild broodstock

Lipids and essential fatty acids, particularly the highly unsaturated fatty acids, 20:5n-3 (eicosapentaenoic acid; EPA), 22:6n-3 (docosahexaenoic acid; DHA) and 20:4n-6 (arachidonic acid, AA) have been shown to be crucial determinants of marine fish reproduction directly affecting fecundity, egg quality, hatching success, larval malformation and pigmentation. In Atlantic cod (Gadus morhua L.) culture, eggs from farmed broodstock can have much lower fertilisation and hatching rates than eggs from wild broodstock. The present study aimed to test the hypothesis that potential quality and performance differences between eggs from different cod broodstock would be reflected in differences in lipid and fatty acid composition. Thus eggs were obtained from three broodstock, farmed, wild/fed and wild/unfed, and lipid content, lipid class composition, fatty acid composition and pigment content were determined and related to performance parameters including fertilisation rate, symmetry of cell division and survival to hatching. Eggs from farmed broodstock showed significantly lower fertilisation rates, cell symmetry and survival to hatching rates than eggs from wild broodstock. There were no differences in total lipid content or the proportions of the major lipid classes between eggs from the different broodstock. However, eggs from farmed broodstock were characterised by having significantly lower levels of some quantitatively minor phospholipid classes, particularly phosphatidylinositol. There were no differences between eggs from farmed and wild broodstock in the proportions of saturated, monounsaturated and total polyunsaturated fatty acids. The DHA content was also similar. However, eggs from farmed broodstock had significantly lower levels of AA, and consequently significantly higher EPA/AA ratios than eggs from wild broodstock. Total pigment and astaxanthin levels were significantly higher in eggs from wild broodstock. Therefore, the levels of AA and phosphatidylinositol, the predominant AA-containing lipid class, and egg pigment content were positively related to egg quality or performance parameters such as fertilisation and hatching success rates, and cell symmetry