Essential fatty acid requirement of juvenile red drum (Sciaenops ocellatus)

Feeding experiments and laboratory analyses were conducted to establish the essential fatty acid (EFA) requirement of red drum (Sciaenops ocellatus). Juvenile red drum were maintained in aquaria containing brackish water (5 ± 2‰ total dissolved solids) for two 6-week experiments. Semipurified diets contained a total of 70% lipid consisting of different combinations of tristearin [predominantly 18:0] and the following fatty acid ethyl esters: oleate, linoleate, linolenate, and a mixture of highly unsaturated fatty acids (HUFA) containing approximately 60% eicosapentaenoate plus docosahexaenoate. EFA-deficient diets (containing only tristearin or oleate) rapidly reduced fish growth and feed efficiency, and increased mortality. Fin erosion and a “shock syndrome” also occurred in association with EFA deficiency. Of the diets containing fatty acid ethyl esters, those with 0.5–1% (n-3) HUFA (0.3–0.6% eicosapentaenoate plus docosahexaenoate) promoted the best growth, survival, and feed efficiency; however, the control diet containing 7% menhaden fish oil provided the best performance. Excess (n-3) HUFA suppressed fish weight gain; suppression became evident at 1.5% (n-3) HUFA, and was pronounced at 2.5%. Fatty acid compositions of whole-body, muscle and liver tissues from red drum fed the various diets generally reflected dietary fatty acids, but modifications of these patterns also were evident. Levels of saturated fatty acids appeared to be regulated independent of diet. In fish fed EFA-deficient diets (containing only tristearin or oleate), monoenes increased and (n-3) HUFA were preferentially conserved in polar lipid fractions. Eicosatrienoic acid [20:3(n-9)] was not elevated in EFA-deficient red drum, apparently due to their limited ability to transform fatty acids. Red drum exhibited some limited ability to elongate and desaturate linoleic acid [18:2(n-6)] and linolenic acid [18:3(n-3)]; however, metabolism of 18:3(n-3) did not generally result in increased tissue levels of (n-3) HUFA. Based on these responses, the red drum required approximately 0.5% (n-3) HUFA in the diet (approximately 7% of dietary lipid) for proper growth and health.     

Requirements for n-3 highly unsaturated fatty acids in feeding juvenile Iranian sturgeon (<Emphasis Type="Italic">Acipenser persicus</Emphasis>) and its effects on growth,carcass quality,and fatty acid composition

A 60-day feeding experiment was carried out on juvenile Iranian sturgeon (Acipenser persicus) to evaluate the effects of different percentages of canola oil and fish oil containing n-3 highly unsaturated fatty acids (n-3 HUFA) on fish growth and fatty acid composition. The requirement for n-3 HUFA of juvenile Iranian sturgeon (48.4 ± 1.98 g) was studied by feeding the fish with various diets containing six different percentage of n-3 HUFA ranging from 1.56 to 17.25 (% of total fatty acids). Neither the weight gain, feed conversion ratio, condition factor, specific growth rate nor the protein efficiency ratio showed any significant differences between the dietary treatments nor in the body composition of juvenile Iranian sturgeon (P > 0.05); also there were no significant difference with respect to the effect of the dietary treatment (P > 0.05) on the blood parameters, for the content of plasma protein, glucose, cholesterol, and triglyceride. The fatty acid composition of the carcass of the Iranian sturgeon fed with the diets containing various levels of n-3 HUFA was reflected by the dietary fatty acid composition. The content of n-3 HUFA in the fish increased with an increase in dietary n-3 HUFA levels. The results indicate that the dietary n-3 HUFA had no effect on the growth of juvenile Iranian sturgeon.     

Incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids in phospholipid classes in cultured turbot (Scophthalmus maximus) cells

The incorporation and metabolism of various (n-3) and (n-6) polyunsaturated fatty acids (PUFA) supplemented to the culture medium was investigated in a turbot cell line (TF). The distribution, and the occurrence and extent of further metabolism of incorporated PUFA via desaturation/elongation mechanisms in specific phospholipid classes was determined from the different fatty acid compositions. The cells contained Δ6 and Δ4 desaturase activities but were generally deficient in C18–20 elongase activity. Δ5 Desaturase activity was generally masked by this deficiency but was present. The compositional data indicated that there was a high degree of specificity between individual phospholipid classes and particular fatty acids probably driven by the specificities of the acylating enzymes. The highest percentages of the supplemented acids were generally observed in the phosphatidic acid/cardiolipin fraction (PA/CL), suggesting a role for PA in the incorporation of the supplemented acids into the phospholipid pool. PI had a characteristic composition consistent with a putative role as a pool of precursor fatty acid for eicosanoid synthesis. Mechanisms were evident for generating and/or maintaining this composition.     

A quantitative comparison between diet and body fatty acid composition in wild northern pike (Esox lucius L.)

The fatty acid compositions of wild female northern pike (Esox lucius L.) and their principle prey species were compared to assess the extent to which pike modify the relative abundance of dietary fatty acids during assimilation and to indicate the optimum dietary content of essential fatty acids (EFAs) for pike. Only minor differences existed between the estimated whole body fatty acid composition of pike and diet fatty acid composition as calculated from the contribution of each prey species to the pike's diet. Saturated fatty acids comprised a slightly higher percentage of diet lipids (25% wt) than of pike lipids (21% wt) whereas monounsaturated fatty acids were less abundant in diet lipids (26% wt) than in pike (29% wt). Percentages of total polyunsaturated fatty acids (PUFAs), n - 3 fatty acids, and n - 6 fatty acids were approximately 43, 30, and 13% wt respectively and differed by less than 1% wt between pike and diet lipids. Among individual PUFAs, the largest differences occurred in 20:5 (n-3) and 22:6(n-3) which comprised, on average, 9.6 and 14.7% wt respectively of diet lipids and 5.9 and 18.3% wt respectively of pike lipids. The close similarity in fatty acid composition between pike and their diet suggests that pike may have limited abilities to elongate and desaturate 18 carbon PUFAs and may require specific long chain PUFAs in the diet. The n-3 PUFA content of the pike's natural diet may exceed the minimum EFA requirements of better studied species such as rainbow trout and turbot.     

Effects of temperature and dietary n-3 and n-6 fatty acids on endocytic processes in isolated rainbow trout (Oncorhynchus mykiss,Walbaum) hepatocytes

Effects of different incubation temperatures (2, 8, 14 and 20°C) and hepatocyte membrane fatty acid composition on the rate of internalization and lysosomal degradation of the ligand, mannosylated albumin, that is taken up by receptor-mediated endocytosis, were investigated in rainbow trout (Oncorhynchus mykiss, Walbaum). The fish were kept at a water temperature ranging from 9 to 14°C and fed pelleted diets coated with either capelin oil (control), EPA/DHA-concentrate (rich in n-3 polyunsaturated fatty acids) or soybean oil (rich in n-6 unsaturated fatty acids) for at least 3 months prior to sampling. The endocytic uptake mediated by the mannose receptor was very efficient at all temperatures studied. Lysosomal degradation, on the other hand, came to a halt below 8°C. The activation energies for uptake and degradation were 54.6 and 164.2 kJ/mol respectively. No negative effects of increased amounts of either n-3 or n-6 fatty acids were observed on the endocytic parameters studied. On the contrary, multivariate analysis indicated a positive relationship between high levels of n-6 fatty acids and low unsaturation index in the phosphatidylcholine (PC) fraction of the hepatocytes and the internalization rate of 2°C, meaning that the rate of receptor-mediated endocytosis may be affected by membrane fatty acid composition.     

Occurrence of all-<Emphasis Type="Italic">cis</Emphasis>-5,8,11,14,17,20,23-hexacosaheptaenoic acid (26:7n-3) in roughscale sole <Emphasis Type="Italic">Clidoderma asperrimum</Emphasis> flesh lipids

Fatty acid analysis of roughscale sole Clidoderma asperrimum flesh lipids was carried out by gas chromatography. An unidentified peak appeared in the chromatogram in the elution region of ≥C24 fatty acids. After enrichment by solvent partitioning, reversed-phase thin-layer chromatography (TLC), and argentation TLC, the peak component was subjected to structural analyses. The partially hydrogenated products after reaction with hydrazine hydrate gave seven isomers of cis-hexacosenoic acid (26:1). Gas chromatography-mass spectrometry (GC–MS) analyses of their dimethyl disulfide adducts identified the monounsaturates as 5-, 8-, 11-, 14-, 17-, 20-, and 23-26:1. The peak component was assigned to all-cis-5,8,11,14,17,20,23-hexacosaheptaenoic acid (26:7n-3). GC–MS analyses of the 4,4-dimethyloxazoline derivative and methyl ester confirmed this structure. This fatty acid is a rare, very long-chain polyunsaturated fatty acid (VLCPUFA). The concentrations of the acid found in roughscale sole were 0.69 ± 0.34% (N = 5) of total fatty acids in flesh lipids. Roughscale sole appears to be characterized by the occurrence of 26:7n-3, which is lacking in popular sources of methylene-interrupted VLCPUFA, such as vertebrate retina, spermatozoa, and herring.     

Determination of amino acid and fatty acid composition of goldband goatfish [Upeneus moluccensis (Bleeker, 1855)] fishing from the Gulf of Antalya (Turkey)

In this study, we aimed to determine the basic food components, fatty acids and amino acids, and variations in these components with months in goldband goatfish (Upeneus moluccensis) that fishing from Gulf of Antalya. As a result of the analyzes, the crude fat values were determined between 1.43 and 3.78%, and the crude protein values were determined between 20.79 and 22.16%. The most abundant fatty acids were determined: palmitic acid (C16:0), stearic acid (C18:0), palmitoleic acid (C16:1c9), oleic acid (C18:1c9), linoleic acid (C18:2n-6), eicosatrienoic acid (C20:3n-3), arachidonic acid (C20:4n-6), eicosapentaenoic acid (C20:5n-3), docosapentaenoic acid (C22:5n-6), and docosahexaenoic acid (C22:6n-3). The most abundant amino acids were determined lysine and leucine, aspartic acid, glutamic acid, alanine, and glycine. The differentiations of essential nutrient components, fatty acids, and amino acids were found generally significant (P < 0.05).

     

Administration of two oils rich in n-3 long-chain polyunsaturated fatty acids to rat pups of dams fed a diet high in fat and low in n-3 polyunsaturated fatty acids

Long-chain polyunsaturated fatty acids (LCPUFA) with 20 or 22 carbons are considered important to the development of infants and sometimes added to infant formulae. In this study, two characteristic sources of n-3 LCPUFA (fish oil and microalgal oil) were orally administrated to rat pups of mildly n-3 PUFA — deficient dams to compare the consequences of the administration. The milk from the dams fed a n-3 PUFA — restricted diet contained less n-3 LCPUFA than that of the dams fed a control diet. Pups were administered 1 mg/g weight of the test oil at the age of 5–7 days. At the age of 7 days, they were sacrificed before or after the administration and fatty acid compositions of the stomach and serum lipid were studied. The administration changed docosahexaenoic acid (DHA; 22:6n−3) levels in the stomach contents and serum lipids with time. Eicosapentaenoic acid (EPA; 20:5n−3) levels increased immediately after the administration of fish oil. The administration of microalgal oil also affected the serum lipid EPA level, in spite of a lack of EPA. In this study, both oils effectively supplemented DHA. Fish oil returned the serum EPA level close to the control value while microalgae oil had little effect.     

Reinvestigation of positional distribution of tetracosahexaenoic acid in triacyl-sn-glycerols of flathead flounder flesh

Positional distribution of fatty acids in triacyl-sn-glycerols (TAG) of the flathead flounder Hippoglossoides dubius has been reinvestigated in order to accurately determine the contents of tetracosahexaenoic acid (24:6n-3) in the sn-1, sn-2, and sn-3 positions. Flesh TAG obtained from three flounders were subjected to stereospecific analysis using a suitable procedure for fish TAG analysis. The 24:6n-3 acid was found in the three positions at the concentrations of 0.3–5.5 mole% (the sn-1 position), 1.6–23.3 mole% (the sn-2 position), and 0.6–8.9 mole% (the sn-3 position). In contrast to a previous analysis, the present analysis revealed that 24:6n-3 is preferentially esterified in the sn-2 position followed by the sn-3 and sn-1 positions. Other polyunsaturated fatty acids, docosahexaenoic acid (22:6n-3; DHA) and docosapentaenoic acid (22:5n-3; DPA), showed a similar distribution pattern. These results indicate that the general tendency observed for long-chain polyunsaturated fatty acids in marine fish TAG can be extended to the distribution of 24:6n-3 in flathead flounder TAG. Because the use of flathead flounders is entirely for human food, we thus intake 24:6n-3 concentrated in the sn-2 position of their TAG.     

Nutritional regulation of hepatocyte fatty acid desaturation and polyunsaturated fatty acid composition in zebrafish (Danio rerio) and tilapia (Oreochromis niloticus)

The desaturation and elongation of [1-¹⁴C]18:3n-3 was investigated in hepatocytes of the tropical warm freshwater species, zebrafish (Danio rerio) and Nile tilapia (Oreochromis niloticus). The hepatocyte fatty acid desaturation/elongation pathway was assayed before and after the fish were fed two experimental diets, a control diet containing fish oil (FO) and a diet containing vegetable oil (VO; a blend of olive, linseed and high oleic acid sunflower oils) for 10 weeks. The VO diet was formulated to provide 1% each of 18:2n-6 and 18:3n-3, and so satisfy the possible EFA requirements of zebrafish and tilapia. At the end of the dietary trial, the lipid and fatty acid composition was determined in whole zebrafish, and liver, white muscle and brain of tilapia. Both zebrafish and tilapia expressed a hepatocyte fatty acid desaturation/elongation pattern consistent with them being freshwater and planktonivorous fish. The data also showed that hepatic fatty acid desaturation/elongation was nutritionally regulated with the activities being higher in fish fed the VO diet compared to fish fed the FO diet. In zebrafish, the main effect of the VO diet was increased fatty acid Δ6 desaturase activity resulting in the production of significantly more 18:4n-3 compared to fish fed the FO diet. In tilapia, all activities in the pathway were greater in fish fed the VO diet resulting in increased amounts of all fatty acids in the pathway, but primarily eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). However, the fatty acid compositional data indicated that despite increased activity, desaturation of 18:3n-3 was insufficient to maintain tissue proportions of EPA and DHA in fish fed the VO diet at the same level as in fish fed the FO diet. Practically, these results indicate that manipulation of tilapia diets in commercial culture in response to the declining global fish oil market would have important consequences for fish fatty acid composition and the health of consumers. Scientifically, zebrafish and tilapia, both the subject of active genome mapping projects, could be useful models for studies of lipid and fatty acid metabolism at a molecular biological and genetic level. This revised version was published online in August 2006 with corrections to the Cover Date.     

Polyunsaturated fatty acid metabolism in cultured fish cells: Incorporation and metabolism of (n-3) and (n-6) series acids by Atlantic salmon (Salmo salar) cells

The incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids (PUFA) supplemented to growing cultures were studied in Atlantic salmon (AS) cells. A fatty acid concentration of 25 μM considerably altered the fatty acid composition of AS cells without increasing the neutral lipid content of the cells or inducing the production of cytoplasmic lipid droplets. Whereas Δ6 and Δ5 desaturase activities were significantly expressed in AS cells, Δ4 desaturase activity was very low. Both the Δ6 desaturase activity and the Δ5 desaturase activity showed some preference for (n-3) PUFA.     

Influence of partial substitution of dietary fish oil by vegetable oils on the metabolism of [1-14C] 18:3n-3 in isolated hepatocytes of European sea bass (Dicentrarchus labrax L.)

The static or declining supply of fish oil from industrial fisheries demands the search of alternatives, such as plant (vegetable) oils, for diets in expanding marine aquaculture. Vegetable oils are rich in C₁₈ polyunsaturated fatty acids but devoid of the n-3 highly unsaturated fatty acids in fish oils. Previous studies, primarily with salmonids, have shown that including vegetable oils in their diets increased hepatocyte fatty acid desaturation. In the present study, we have investigated the effects of dietary partial substitution of fish oil (FO) with rapeseed oil (RO), linseed oil (LO) and olive oil (OO) on the desaturation /elongation and, -oxidation capacities of [1-¹⁴C]18:3n-3 in isolated hepatocytes from European sea bass (Dicentrarchus labrax L.), in a simultaneous combined assay. Fish were fed during 34 weeks with diets containing 100% FO, or RO, LO and OO, each included at 60% with the balance being met by FO, with no detrimental effect upon growth or survival. The highest total desaturation rates were found in hepatocytes of fish fed FO diet (0.52±0.08 pmol/h/mg protein) and OO diet (0.43±0.09 pmol/h/mg protein), which represented 3.2% and 2.7% of total [1-¹⁴C]18:3n-3 incorporated, respectively. In contrast, lowest desaturation rates were presented by hepatocytes of fish fed LO and RO diets (0.23±0.06 and 0.14±0.05 pmol/h/mg protein, respectively) represented 1.4% and 0.9% of total [1-¹⁴C]18:3n-3 incorporated, respectively. The rates of [1-¹⁴C]18:3n-3 β-oxidized were between 11-fold and 35-fold higher than desaturation. However, no significant differences were observed among β-oxidation activities in hepatocytes of fish fed any of the diets. The present study demonstrated that the European sea bass, as a carnivorous marine fish, presented a ‘marine’ fish pattern in the metabolism of 18:3n-3 to 20:5n-3 and 22:6n-3. This species appeared to have all the enzymic activities necessary to produce 22:6n-3 but presented only extremely low rates of fatty acid bioconversion. Furthermore, nutritional regulation of hepatocyte fatty acid desaturation was minimal, and dietary vegetable oils did not increase desaturase activities, and in RO and LO treatments the activity was significantly lower. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ration of 20:3 (n-9) to 20:5 (n-3) in Phospholipids as an Indicators of Dietary Essential Fatty Acid Sufficiency in Striped Bass,Morone saxarilis,and Palmetto Bass,M. saxatilis x M. chrysops

The effects of feeding different sources of brine shrimp nauplii with different fatty acid compositions on growth, survival, and fatty acid composition of striped bass, Morone saxarilis and palmetto bass (M. saxatilis x M. chrysops) were determined. The sources of brine shrimp were Chinese (CH), with a high percentage of 20:5(n-3), eicosapentaenoic acid (EPA), and Colombian (COL), San Francisco Bay (SFB), and Great Salt Lake (GSL), with low percentages of EPA but high percentages of 18:3(n-3), linoienic acid. None of the brine shrimp sources contained a measurable amount of 22:6(n-3), docosahexaenoic acid (DHA). After enrichment with menhaden oil to increase the content of EPA and DHA, the GSL brine shrimp nauplii were also fed to hybrid striped bass.Growth and survival of fish larvae fed brine shrimp nauplii with high percentages of EPA and DHA (CH and GSLE) were higher (P < 0.05) than those of fish fed brine shrimp with a low percentage of EPA (COL, SFB, and GSL). The ratio of 20:3(n-9) eicosatrienoic acid (ETA), to DHA in polar lipids (phospholipids) of fish, traditionally used as an indicator of essential fatty acid (EFA) sufficiency of the diet, was not a reliable indicator of essential fatty acid sufficiency of diets for larval striped bass and hybrid striped bass. However, the ratio of ETA to EPA appears to be an appropriate indicator. An ETA-to-EPA ratio in phospholipids of less than 0.10 is consistent with an EFA sufficient diet.     

n-3 LC-PUFA deposition efficiency and appetite-regulating hormones are modulated by the dietary lipid source during rainbow trout grow-out and finishing periods

Largely attributable to concerns surrounding sustainability, the utilisation of omega-3 long-chain polyunsaturated fatty acid-rich (n-3 LC-PUFA) fish oils in aquafeeds for farmed fish species is an increasingly concerning issue. Therefore, strategies to maximise the deposition efficiency of these key health beneficial fatty acids are being investigated. The present study examined the effects of four vegetable-based dietary lipid sources (linseed, olive, palm and sunflower oil) on the deposition efficiency of n-3 LC-PUFA and the circulating blood plasma concentrations of the appetite-regulating hormones, leptin and ghrelin, during the grow-out and finishing phases in rainbow trout culture. Minimal detrimental effects were noted in fish performance; however, major modifications were apparent in tissue fatty acid compositions, which generally reflected that of the diet. These modifications diminished somewhat following the fish oil finishing phase, but longer-lasting effects remained evident. The fatty acid composition of the alternative oils was demonstrated to have a modulatory effect on the deposition efficiency of n-3 LC-PUFA and on the key endocrine hormones involved in appetite regulation, growth and feed intake during both the grow-out and finishing phases. In particular, n-6 PUFA (sunflower oil diet) appeared to ‘spare’ the catabolism of n-3 LC-PUFA and, as such, resulted in the highest retention of these fatty acids, ultimately highlighting new nutritional approaches to maximise the maintenance of the qualitative benefits of fish oils when they are used in feeds for aquaculture species.     

Importance of fatty acids in broodstock diets with emphasis on Arctic char (Salvelinus alpinus) eggs

The importance of long-chain polyunsaturated fatty acids, especially the eicosanoid precursors, is addressed in this paper. It has been generally recognized that eicosapentaenoic (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) are of significant importance in fish reproduction while arachidonic acid (AA, 20:4n-6) has often been overlooked. The ratio between C20 fatty acids EPA and AA might be important for many physiological functions depending on the species evolution and its requirements. Arctic char (Salvelinus alpinus) has a much more pronounced freshwater history and therefore different fatty acid requirements than the other commonly farmed salmonids such as salmon (Salmo salar), brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss). Therefore there is reason to formulate a feed that is more suitable for farming of this freshwater species. In this study, freshwater wild-origin char eggs were compared to farmed eggs of char. The ratio n-3/n-6 of total phospholipids of eggs was much lower in the wild fish, 3.5 versus 13.5, and the hatching rate of eggs from natural environment was much higher (20–70% vs. >80%). We conclude that feed based on marine raw product does not fulfill the requirements for essential fatty acids for freshwater char and we suggest that AA is supplemented to the broodstock diet and that at least linoleic acid (18:2n-6) is included in the on-growth diet formulas to lower the n-3/n-6 fatty acid ratio.     

Effects of dietary lipid level and vegetable oil on fatty acid metabolism in Atlantic salmon (Salmo salar L.) over the whole production cycle

Changes in fatty acid metabolism in Atlantic salmon (Salmo salar) induced by vegetable oil (VO) replacement of fish oil (FO) and high dietary oil in aquaculture diets can have negative impacts on the nutritional quality of the product for the human consumer, including altered flesh fatty acid composition and lipid content. A dietary trial was designed to investigate the twin problems of FO replacement and high energy diets in salmon throughout the entire production cycle. Salmon were grown from first feeding to around 2 kg on diets in which FO was completely replaced by a 1:1 blend of linseed and rapeseed oils at low (14–17%) and high (25–35%) dietary oil levels. This paper reports specifically on the influence of diet on various aspects of fatty acid metabolism. Fatty acid compositions of liver, intestinal tissue and gill were altered by the diets with increased proportions of C₁₈ polyunsaturated fatty acids and decreased proportions of n-3 highly unsaturated fatty acids (HUFA) in fish fed VO compared to fish fed FO. HUFA synthesis in hepatocytes and enterocytes was significantly higher in fish fed VO, whereas β-oxidation was unaltered by either dietary oil content or type. Over the entire production cycle, HUFA synthesis in hepatocytes showed a decreasing trend with age interrupted by a large peak in activity at seawater transfer. Gill cell prostaglandin (PG) production showed a possible seasonal trend, with peak activities in winter and low activities in summer and at seawater transfer. PG production in seawater was lower in fish fed the high oil diets with the lowest PG production generally observed in fish fed high VO. The changes in fatty acid metabolism induced by high dietary oil and VO replacement contribute to altered flesh lipid content and fatty acid compositions, and so merit continued investigation to minimize any negative impacts that sustainable, environmentally-friendly and cost-effective aquaculture diets could have in the future. Abbreviations: FO - fish oil; HUFA - highly unsaturated fatty acids acids (carbon chain length ≥C ₂₀ with ≥3 double bonds); LO - linseed oil; RO - rapeseed oil; VO - vegetable oil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative fatty acid profiles of wild and farmed tropical freshwater fish rohu (<Emphasis Type="Italic">Labeo rohita</Emphasis>)

The proximate composition of the whole body and the fatty acid composition of the liver, muscle, eye and brain of wild and cultured rohu (Labeo rohita) were analyzed. The cultured species was found to have significantly (P < 0.05) higher lipid contents than its wild counterpart. The saturated (SFA) and monounsaturated (MUFA) fatty acid contents were significantly higher in the cultured species, whereas the n-6 and n-3 polyunsaturated fatty acid (PUFA) levels were higher in the wild species. Fatty acids C16:0 and C18:1 n-9 were the principal fatty acids of the SFAs and MUFAs, respectively, identified in the analyses. Docosahexaenoic acid, eicosapentaenoic acid, and arachidonic acid were the predominant PUFAs in both groups, and all three were found to be present at significantly (P < 0.05) higher levels in the wild species. Erucic acid (C22:1 n-9), which was the predominant fatty acid (30.76%) in the feed, was detected only at low levels in muscle (0.30%), liver (1.04%) and eye (1.28%) of cultured fish tissue.     

Effect of soybean oil and soybean lecithin on intestinal lipid composition and lipid droplet accumulation of rainbow trout, Oncorhynchus mykiss Walbaum

Rainbow trout (Oncorhynchus mykiss Walbaum) were fed purified diets containing fish oil for six weeks and then soybean lecithin or soybean oil for 25 days. The gastrointestinal tract segments, stomach, midgut and hindgut were then sampled for lipid and fatty acid composition and electron microscopy. Membrane lipid class composition was fairly similar in all three segments of trout fed fish oil. Hindgut contained slightly more phosphatidylserine than stomach and midgut, while midgut contained more phosphatidylcholine and less lysophospatidylcholine/sphingomyelin. Feeding soybean products appeared to marginally decrease free cholesterol. The fatty acid compositions of the main lipid classes showed significant regional differences. In control fish, stomach had higher levels of arachidonic acid (20:4n-6) and n-6 polyunsaturated fatty acids than midgut and hindgut, and lower content of docosahexaenoic acid (22:6n-3) and n-3 polyunsaturated fatty acids. Midgut phosphatidylethanolamine also had higher levels of saturated fatty acids and less n-3 polyunsaturated fatty acids than the other tissues. Feeding soybean products decreased the n-3/n-6 ratio mainly due to increases in linoleic (18:2n-6) and 20:4n-6 and decreases in 22:6n-3 and eicosapentaenoic acid (20:5n-3). Phosphatidylcholine and to a lesser extent phosphatidylethanolamine adapted more readily to dietary changes by major increases in 18:2n-6 and C₂₀₋₂₂ n-6 polyunsaturated fatty acids. The composition of phosphatidyl-serine and -inositol appeared to be under more strict metabolic control. Linoleic acid hardly increased at all while the increase in other n-6 polyunsaturated fatty acids was less pronounced than for the other lipid classes. Regardless of lipid class, stomach resisted dietary changes more strongly than midgut and hindgut. Increases in n-6 polyunsaturated fatty acids were minor as were the loss of n-3 polyunsaturated fatty acids. The dead-end product 20:2n-6 accumulated to a higher degree in hindgut phosphatidyl-ethanolamine and -coline compared to midgut and stomach, suggesting that the activity of Δ₆ desaturation is higher in the anterior part of the intestine where most of the lipid is absorbed. Feeding soybean oil caused massive accumulation of free lipid droplets in midgut enterocytes while little lipid droplets were observed in trout fed fish oil or soybean lecithin. Since both soybean products influenced intestinal composition to the same degree, altered fatty acid profiles in membranes is not responsible for the observed lipid accumulation. This supports previous observations in Arctic charr (Salvelinus alpinus L.), suggesting that fish may require exogenous phospholipids in order to sustain a sufficient rate of lipoprotein synthesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of dietary n-3 fatty acids on the desaturation and elongation of [1-14C] 18:2 n-6 and [1-14C] 18:3 n-3 in Atlantic salmon hepatocytes

Atlantic salmon (Salmo salar) were fed diets containing fish oil supplemented with 22:6n-3 (FO diet) or linseed oil supplemented with 20:5n-3 (LO diet) for 6 months. The effects of these diets, both containing about 36% n-3 fatty acids, on the esterification, desaturation and elongation of [1-¹⁴C] 18:2n-6 and [1-¹⁴C] 18:3n-3 were investigated in isolated hepatocytes. The percentages of radioactivity which was esterified from [1-¹⁴C] 18:2n-6 or [1-¹⁴C]18:3n-3 into total lipids, were approximately 20% lower in hepatocytes from fish fed the FO diet than in hepatocytes from fish fed the LO diet. The percentages of radioactivity esterified in both groups were further reduced when 0.1 mM unlabelled 22:6n-3 was added to the incubation. The percentage of desaturation and elongation products formed from [1-¹⁴C] 18:2n-6 was twice as high in hepatocytes from salmon fed the FO diet as it was in hepatocytes from fish fed the LO diet. The ratio of 18:2n-6 to 18:3n-3 was five times higher in the FO diet, and this probably promoted the conversion of 18:2n-6 to longer chain n-6 fatty acids. When 0.1mM unlabelled 22:6n-3 was added to the incubation medium, the percentages of desaturation and elongation products formed were unchanged. Thus, a high level of 22:6n-3 in the diet is apparently not inhibiting the conversion of 18:2n-6 to 20:4n-6, as long as the amount of 18:2n-6 present is substantially higher than that of 18:3n-3. No desaturation and elongation products were recovered from the phospholipids of hepatocytes incubated with [1-¹⁴C] 18:3n-3 in any of the groups. However, the `dead end' elongation product 20:3n-3 was found in the triacylglycerol fraction, and the percentage of this fatty acid increased when 22:6n-3 was added to the incubation medium.     

Incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids in phospholipid classes in cultured rainbow trout (Salmo gairdneri) cells

The incorporation and metabolism of various (n-3) and (n-6) polyunsaturated fatty acids supplemented to the culture medium was investigated in the rainbow trout cell line, RTG-2. The distribution, and the occurrence and relative extent of further desaturation and elongation of the incorporated acids was determined in individual phospholipid classes by analysis of the fatty acid compositions. RTG-2 cells exhibited Δ6 and Δ5 desaturase activities whereas Δ4 desaturase activity was almost totally absent. The percentage of precursor acids was greatest in the phosphatidic acid/cardiolipin fraction (PA/CL), suggesting a role for possibly PA in the initial incorporation of these acids into the phospholipid pool. The compositional data indicated that individual intermediates and products of the desaturation pathways were associated with specific phospholipid classes probably via mechanisms depending upon the specificities of the acylating enzymes. The composition of phosphatidylinositol (PI) and the tightly controlled mechanisms for generating/maintaining it are consistent with a role for this phospholipid in providing precursor fatty acid for eicosanoid synthesis.