The desaturation and elongation of 14C-labelled polyunsaturated fatty acids by pike (Esox lucius L.) in vivo

To examine the ability of pike (Esox lucius L.) to modify exogenous PUFA by desaturation and elongation, ¹⁴C-labelled 18:2(n-6), 18:3(n-3), 20:4(n-6) and 20:5(n-3) were injected intraperitoneally and the distribution of radioactivity in tissue lipid classes and liver PUFA measured. In all tissues examined, radioactivity from all ¹⁴C-PUFA was recovered in many classes of acyl lipids and the level of recovery generally reflected the relative abundance of the lipid classes. Triacylglycerols, CGP and EGP usually contained high levels of all incorporated ¹⁴C-PUFA. PI contained higher levels of radioactivity from ¹⁴C-20:4(n-6) than from other injected substrates. In liver lipid, the 6 desaturation products of ¹⁴C-18:2(n-6) and ¹⁴C-18:3(n-3) contained no measurable radioactivity although the elongation products of the 6 desaturation products were labelled, as were the direct elongation products of these injected substrates. No radioactivity from ¹⁴C-18:2(n-6) or ¹⁴C-18:3(n-3) was detected in C₂₀ or C₂₂ products of 5 and 4 desaturation. Almost all radioactivity from injected ¹⁴C-20:4(n-6) was recovered in this PUFA. Of the total radioactivity from ¹⁴C-20:5(n-3) incorporated into liver lipid, 7% was present as 24:5 and 16.4% was recovered in hexaenoic fatty acids. In liver, 24:5(n-3) and 24:6(n-3) each accounted for 1% of the mass of total fatty acids and were located almost exclusively in triacylglycerols. The presence of radioactivity in these C₂₄ PUFA suggests that in pike the synthesis of 22:6(n-3) from 20:5(n-3) may proceed without 4 desaturase via the pathway which involves chain shortening of 24:6(n-3). It is concluded that under the circumstances employed in this study pike, do not exhibit 5 desaturase activity and are unable to synthesize 20:4(n-6) and 20:5(n-3) from 18:2(n-6) and 18:3(n-3), respectively. This suggests that pike may require 20:4(n-6) and 20:5(n-3) preformed in the diet.Abbreviations CGP choline glycerophospholipids - CL cardiolipin - EGP ethanolamine glycerophospholipids - PG phosphatidylglycerol - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acids - SM sphingomyelin - TLC thin-layer chromatography     

河川沙塘鳢胚胎、仔鱼发育过程中脂类含量及脂肪酸组成的变化

采用生物化学方法测定和分析了河川沙塘鳢(Odontobutis potamophila)胚胎、仔鱼发育过程中脂类的含量及脂肪酸的组成。结果显示,随着胚胎和仔鱼的发育,其总脂的含量呈下降趋势,饱和脂肪酸(SFA)的含量亦呈现出下降趋势,单不饱和脂肪酸(MUFA)的含量在胚胎期保持稳定、在仔鱼期下降,而多不饱和脂肪酸(PUFA)却呈现出不断增长的趋势。在整个胚胎和仔鱼发育过程中,平均含量最高的脂肪酸依次是C16∶0、C18∶1n-9、C18∶0、C16∶1、C22∶6n-3(DHA)和C20∶5n-3(EPA)。结果表明:河川沙塘鳢在胚胎和仔鱼发育过程中有消耗饱和脂肪酸和单不饱和脂肪酸,而保存n-3系列和n-6系列的高度不饱和脂肪酸的趋势,饱和脂肪酸被作为胚胎期能量代谢的主要来源,单不饱和脂肪酸被作为仔鱼期能量代谢的主要来源,而C18∶3n-3(亚麻酸)和C18∶2n-6(亚油酸)被用于合成DHA、EPA和C20∶4n-6(AA)。     

Polyunsaturated fatty acid metabolism in a cell culture model of essential fatty acid deficiency in a freshwater fish, carp (Cyprinus carpio)

Proliferation of an essential fatty acid deficient cell line from carp (EPC-EFAD; epithelioma papillosum carp-essential fatty acid deficient) is stimulated by supplementing the cells with C₂₀, but not C₁₈ polyunsaturated fatty acids (PUFA). It is hypothesized that the differential ability of the PUFA to stimulate proliferation of the EPC-EFAD cells may be related to the extent of the cells' ability to desaturate and elongate C₁₈ PUFA. In the present study, the metabolism of ¹⁴C-labeled C₁₈ and C₂₀ PUFA was investigated in EPC-EFAD cells in comparison with normal EPC cells. The incorporation of all the PUFA was significantly greater in EPC-EFAD cells but the rank order, 20:5n-3 > 18:3n-3 = 18:2n-6 >20:4n-6 was the same in both cell lines. The proportion of radioactivity from all labeled PUFA recovered in phosphatidylethanolamine and total polar lipids was significantly lower in EPC-EFAD cells compared to EPC cells, whereas the proportion of radioactivity recovered in all the other phospholipid classes and total neutral lipid was greater in EPC-EFAD cells. Both cell lines desaturated[1-¹⁴C]18:3n-3 and [1-¹⁴C]20:5n-3 to a greater extent than the corresponding (n-6) substrates but the desaturation of all the ¹⁴ C-labeled PUFA was significantly greater in EPC-EFAD cells compared to EPC cells. The results showed that, although essential fatty acid deficiency had several significant effects on PUFA metabolism in EPC cells, the fatty acid desaturation/elongation pathway was not impaired in EPC-EFAD cells and so they can desaturate 18:3n-3 to 20:5n-3 and 22:6n-3, and 18:2n-6 to 20:4n-6. However, 20:4n-3 and 20:3n-6, and not 20:4n-6 and 20:5n-3, were the predominant C₂₀ PUFA produced by the elongation and desaturation of [1-¹⁴C]18:3n-3 and [1-¹⁴C]18:2n-6, respectively. Therefore, the previously reported inability of 18:3n-3 and 18:2n-6, compared to 20:5n-3 and 20:4n-6, to stimulate proliferation of the cells is apparently not due to a general deficiency in the fatty acid desaturation pathway in EPC-EFAD cells but may be related to potential differences in eicosanoid profiles in cells supplemented with C₁₈ PUFA compared to C₂₀ PUFA.     

The effect of culture on morphology, lipid and fatty acid composition, and polyunsaturated fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells

Rainbow trout (Oncorhynchus mykiss) skin cell cultures were obtained by trypsinization of the tissue and grown in Leibovitz L-15 medium. Lipid class compositions, and fatty acid profiles of total lipids and individual phospholipid classes were determined at different times of culture. The metabolism of polyunsaturated fatty acids (PUFA) was investigated by incubating primary cultures after 7 and 14 days with [1-¹⁴C]18:2n-6 and [1-¹⁴C-]18:3n-3. The change in morphology between epithelial-like primary cultures and fibroblastic-like secondary subcultures was accompanied by alterations in the lipid composition. Polar lipids became predominant by 14 days in culture. The relative proportions of phosphatidylcholine (PC), the most abundant phospholipid, phosphatidylinositol and cholesterol increased significantly, while sphingomyelin decreased. Saturated fatty acids, 18:1n-9, n-6 and n-9PUFA were more abundant in total lipid in cultures at 14 days and 4 months than in cells initially isolated which contained higher percentages of longer chain monoenes and n-3PUFA. The changes in fatty acid composition with time in culture were observed in all the major phospholipid classes. Rainbow trout skin cells in culture desaturated and elongated both 18:2n-6 and 18:3n-3, with 20:4n-6 and 20:5n-3 being the most abundant products, respectively. PC presented the highest incorporation of radioactivity, especially following incubation with 18:3n-3. Lipid metabolism in general increased with the age of primary cultures, with both the amount of C18 PUFA incorporated and metabolized by desaturation/elongation significantly increased in 14 day cultures compared to 7 day cultures. Product/precursor ratios calculated for both n-6 and n-3 fatty acids showed that, while 6 desaturase activity was increased significantly with cell age, 5 desaturase activity was more affected by the fatty acid series, with 18:3n-3 being more readily transformed to 20:5n-3 than 18:2n-6 to 20:4n-6. Further desaturation of 20:5n-3 to hexaenes was low. Overall, the data suggested that the trout skin cell cultures were more similar to mammalian skin fibroblasts than mammalian epidermal/keratinocyte cultures.     

The conversion of linoleic acid and linolenic acid to longer chain polyunsaturated fatty acids by Tilapia (Oreochromis) nilotica in vivo

Tilapia (Oreochromis) nilotica were fed either a commercial diet containing 2.2% (n-3) and 0.5% (n-6) polyunsaturated fatty acids (PUFA), or a diet containing 1.0% methyl linoleate as the only PUFA. The fatty acid composition of tissue lipids generally reflected that of the diet. Fish from both dietary groups were injected intraperitoneally with ¹⁴C-labelled linoleic acid, 18:2 (n-6), or linolenic acid, 18:3 (n-3), and the distribution of radioactivity in tissue lipids examined. The conversion of both 18:2 (n-6) and 18:3 (n-3) to longer chain PUFA was lower in fish fed the commercial diet than in those fed the diet containing only 18:2 (n-6). Half of the radioactivity from both substrates recovered in liver polar lipids was present in C20 and C22 PUFA with fish maintained on the experimental diet. It is concluded that T. nilotica is capable of elongating and desaturating both 18:2 (n-6) and 18:3 (n-3), but that this conversion is suppressed by dietary longer chain PUFA. NERC Unit of Aquatic Biochemistry     

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.     

Essential fatty acids in Atlantic salmon: time course of changes in fatty acid composition of liver, blood and carcass induced by a diet deficient in n-3 and n-6 fatty acids

Atlantic salmon fry (4 g) were fed a semi-synthetic diet devoid of n-3 and n-6 polyunsaturated fatty acids (PUFA) for 4 months. External signs of nutritional imbalance during the period were slow growth rate and increased mortality. Some symptoms of essential fatty acid deficiency appeared in the fatty acid composition of the blood and liver during the fourth month. At that time, the percentages of n-3 and n-6 PUFA of triacylglycerols (TAG) were nearly exhausted, and the percentages of 22:6 n-3 and of 20:5 n-3 in phospholipids (PL) showed a marked decrease. This decrease in the PUFA level of the PL was paralleled by the appearance of 20:3 n-9, whereas in the TAG an increase in the percentage of 18:1 n-9 was observed. After 4 months the monounsaturated fatty acid 18:1 n-9 constituted nearly 50% of the fatty acids in the TAG fraction of the liver. The time course of the changes in fatty acid composition of liver and blood lipids was quite similar, whereas the carcass lipid composition appeared to respond slowly to a diet devoid of essential fatty acids.     

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.     

The digestion and selective absorption of dietary fatty acids in Arctic charr, Salvelinus alpinus

To compare the rates of digestion and absorption of individual fatty acids, Arctic charr, Salvelinus alpinus (L.), were fed isoenergetic diets containing 40 g kg^?1 coconut oil, and various combinations of 10 g kg^?1 of polyunsaturated fatty acids (PUFA) (18:2n-6 or 18:3n-3) and monounsaturated fatty acids (MONO) (20:1n-9 or 22:1n-9) in the form of free fatty acids (FFA) or triacylglycerol (TAG). The average lipid digestibility for all diets measured by use of the chromic oxide method in the pyloric caeca area, midgut and hindgut were 72%, 83% and 88%, respectively, showing that lipid digestion and absorption occur mainly in the pyloric caeca area, but also extend throughout the intestinal tract. Analyses of digesta present in the intestinal segments suggest the predominance of non-specific lipolytic activity producing primarily FFA and glycerol from dietary TAG. Comparisons of the fatty acid composition of the lipid classes in the digesta showed that the utilization of dietary TAG was dependent both on the rate of release of the individual fatty acids from TAG, and their subsequent rate of absorption. When supplied as either FFA or TAG, the levels of PUFA (18:2n-6 or 18:3n-3) in the digesta were very low, indicating almost complete utilization. Both MONO used (20:1n-9 or 22:1n-9) were absorbed less efficiently than PUFA, but the rate of release from TAG seemed to be rate limiting only for 22:1n-9, which accumulated in the digesta. The rates of absorption of 20:1n-9 and 22:1n-9 when fed as FFA were the same. Comparisons of the levels of fatty acids in the dietary coconut oil TAG with those of the digesta lipids showed that 12:0 was a good substrate for intestinal lipase and was quickly absorbed. The lipolysis of 14:0 and 16:0 was intermediate while the longer-chain 18:0 appeared very resistant to digestion and was a major component of TAG, diacylglycerols and monoacylglycerols present in particularly the hindgut digesta. The absorption of 18:0 also appeared to be very low. The results suggest that PUFA are released very rapidly from dietary TAG by intestinal lipases in Arctic charr, and are specifically absorbed compared with long-chain saturated and monounsaturated fatty acids. The rate of lipolysis may be the rate-limiting step in the digestion of very long chain monounsaturated fatty acids such as 22:1n-9, while both the rate of lipolysis and absorption may be rate limiting for long-chain saturated fatty acids such as 18:0.     

Essential fatty acids in Atlantic salmon: effects of increasing dietary doses of n-6 and n-3 fatty acids on growth, survival and fatty acid composition of liver, blood and carcass

Atlantic salmon fry (4 g) were fed for 4 months on semi-synthetic diets containing fatty acid methyl esters of either 18:2 n-6, 18:3 n-3 or a mixture of equal amounts of 20:5 n-3 and 22:6 n-3. The different amounts of polyunsaturated fatty acids added were 0, 0.1, 0.2, 0.5, 1 and 2% (by dry weight). Increasing levels of dietary n-3 fatty acids up to 1% gave faster growth rates in salmon fry, and fish fed the mixture of 20:5 n-3 and 22:6 n-3 seemed to grow faster than fish fed only 18:3 n-3. No significant effect on growth rate was seen when the dietary level of 18:2 n-6 was increased. Dietary inclusions of n-3 fatty acids reduced the mortality of salmon, while dietary 18:2 n-6 had no such beneficial effects.
The dietary treatments caused substantial changes in the fatty acid composition of blood and liver phospholipids (PL), whereas the total lipid fraction of the carcass was less affected. Increasing doses of 18:2 n-6 in the diet resulted in an increased percentage of 20:4 n-6 in liver and blood PLs, while the percentage of 20:3 n-9 decreased. The percentage of 18:2 n-6 also increased in liver, blood and carcass. Dietary 18:3 n-3 resulted in increased percentages of 18:3 n-3 and 20:5 n-3 in liver PLs, while the percentage of 20:3 n-9 decreased. There was, however, no significant increase in the percentage of 22:6 n-3. Dietary 18:3 n-3 produced no significant changes in the composition of blood fatty acids, but increased the percentage of 18:3 n-3 in the carcass. The dietary combination of the n-3 fatty acids 20:5 and 22:6 resulted in an increased percentage of 22:6 n-3 in blood and liver lipids and a decreased percentage of 20:3 n-9, but there were no changes in the percentage of 20:5 n-3.     

Dynamics of Total Body Fatty Acids During Early Ontogeny of Pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>) Larvae

The fatty acid composition of pikeperch (Sander lucioperca) was determined according to their physiological status, during starvation (10 days) and feeding (28 days). In starved larvae, polyunsaturated, monounsaturated and saturated fatty acids were utilized as metabolic substrates until day 9. At day 10, all fatty acid levels remained stable or, at least, increased in larval body. Among fatty acids, docosahexaenoic acid 22:6 n-3 was used preferentially (20.3% from total fatty acids utilized) followed by palmitoleic acid 16:1 n-7 (13.9%) and then by oleic 18:1 n-9 (12.3%), linoleic 18:2 n-6 (10.1%), linolenic 18:3 n-3 (9.7%) and eicosapentaenoic 20:5 n-3 (9.1%) acids. On the other hand, arachidonic acid 20:4 n-6 was utilized very lowly (0.3%). In fed larvae, no utilization of body fatty acids was observed during the experiment. It seems that energy requirements (and others) of fed larvae were satisfied by feed.     

Effects of essential fatty acid-deficient diets on growth, mortality, tissue histopathology and fatty acid compositions in juvenile turbot (Scophthalmus maximus)

Three diets in which the lipid component was supplied either as fish oil (FO), linseed oil (LO) or olive oil (OO) were fed to duplicate groups of juvenile turbot (Scophthalmus maximus) of initial weight 1.2 g for a period of up to 12 weeks. The latter two diets resulted in a significant reduction in specific growth rate and an increased mortality compared to the FO (control) fed fish. A liver histopathology was evident in around half of the fish fed the LO and OO diets but was absent in fish fed FO. The lesion showed indications of cellular alterations consisting of foci of densely basophilic cells but without evidence of inflammatory activity. The total lipid fatty acid composition of the carcass from fish fed LO had increased percentages of 18:2n-6 and 18:3n-3, but decreased percentages of all other polyunsaturated fatty acids (PUFA) including the physiologically important 20:4n-6, 20:5n-3 and 22:6n-3, compared to fish fed FO. Almost 2/3 of the total fatty acids in the carcass of OO-fed fish were monounsaturated while the percentages of total saturated fatty acids and all other PUFA, except 18:2n-6, were significantly reduced compared to fish fed FO. Broadly similar effects on total lipid fatty acid composition were observed in liver. In the liver glycerophospholipid classes of fish fed LO, percentages of 18:2n-6, 18:3n-3 and 20:3n-3 were significantly increased whereas all C20 and C22 PUFA, with the exception of 20:5n-3 in PI, were significantly reduced compared to fish fed FO. The liver glycerophospholipids of fish fed OO all showed significantly increased total monounsaturates, 18:2n-6, 20:2n-6, 18:2n-9 and 20:2n-9 as well as reduced percentages of 20:4n-6 and 22:6n-3, compared to fish fed FO. The brain glycerophospholipids showed broadly similar changes in response to dietary treatment although the magnitude of fatty acid alterations was less than those observed in liver. The greater mortalities in the OO-fed fish compared to the LO-fed fish suggests that incorporation of 18:3n-3 into tissue phospholipids can offset losses of long-chain PUFA more effectively than incorporation of 18:1n-9. However, levels of dietary long-chain PUFA must be optimised to allow normal growth and development. We conclude that the very low flux through the fatty acid desaturase/elongase pathways in turbot is not up-regulated by diets deficient in 20:5n-3 and 22:6n-3.     

The effect of dietary n-3 and n-6 fatty acid balance on the growth of the prawn Penaeus monodon

A post-hoc study of the influence of dietary fatty acids of the n-3 and n-6 series on the growth of the prawn, Penaeus monodon showed a clear example of interaction by these nutrients to influence growth. Data from three independent growth studies examining the dietary requirements for linoleic (LOA, 18:2n-6), linolenic (LNA, 18:3n-3), arachidonic (ARA, 20:4n-6), eicosapentaenoic (EPA, 20:5n-2) and docosahexaenoic (DHA, 22:6n-3) acids were standardized through a common reference to allow comparison. Analysis of the variation within the experiments was able to define effects attributable to the individual experiments or the overall dietary n-3 and n-6 levels. A generalized additive model (GAM) indicated that both parameters (experiment, and n-3 and n-6 levels) had significant ( P  < 0.05) effects on growth. Loess nonparametric modelling of the data clearly demonstrates `the effect of relationship' on prawn growth to the levels of dietary n-3 and n-6 fatty acids. The response surface model shows clear effects of both n-3 and n-6, and that the effect of n-3 changes with the level of n-6 (and vice versa). Parametric examination of the relationship ( y =–37.149 x ³ + 160.84 x ² – 118.64 x  + 290.6, r ²=0.492, P  < 0.05) between growth and the ratio between the two fatty acid classes suggested that the optimal ratio of n-3 and n-6 fatty acid is about 2.5 to 1. The results of this study demonstrated that the interaction of the dietary n-3 and n-6 fatty acid classes is an important factor of prawn fatty acid nutrition.     

Chemical changes in fed and starved larval trout cod, Maccullochella macquarensis during early development

The changes in proximate composition, amino acid (total and free) and fatty acid content of artificially propagated trout cod, Maccullochella macquariensis larvae from five mothers hatched, weaned and reared separately, each in two groups, one fed with Artemia naupli and the other starved, for 15 days (after yolk resorption), are presented. There was no significant change in the proximate composition of fed larvae with devlopment, but in starved larvae the protein (linearly) and lipid (curvi-linearly) content decreased significantly as starvation progressed. The essential amino acids (EAA) and non- essential amino acids (NEAA) found in highest amounts in trout cod larvae were lysine, leucine, threonine and arginine, and alanine, serine and glutamic acid, respectively. In fed larvae the total amino acid (TAA), TEAA and TNEAA content did not vary significantly as development progressed. In starved larvae the TAA, EAA and NEAA content, as well as all the individual amino acids decreased significantly (P<0.05) from the levels in day of hatch and/or yolk-sac resorbed larvae. The greatest decrease occurred in the TEAA content (7.38±0.76 at day of hatch to 1.96±0.09 15 day starved in moles larva^–1; approximately a 74% decrease), whereas the decrease in TNEAA was about 38%. Unlike in the case of TAA distinct changes in the free amino acid (FAA) pool were discernible, from day of hatch and onwards, in both fed and starved trout cod larvae. In both groups of larvae the most noticeable being the decrease of % FEAA in TFAA, but not the % FAA in TAA. Four fatty acids together, accounted for more than 50% of the total in each of the major fatty acid categories in all larvae sampled; 16: 0, 18:1n-9, 22: 6n-3 and 20: 4n-6, amongst saturates, monoenes, n-3 PUFA and n-6 PUFA, respectively. Twelve fatty acids either decreased (14: 0, 16: 1n-7, 20: 1n-9, 20: 4n-6, 20: 5n-3, 22: 5n-3 and 22: 6n-3) or increased (18: 2n-6, 18: 3n-3, 18: 3n-6, 18: 4n-3 and 20: 3n-3) in quantity, after 15 days of feeding, from the base level in day of hatch and/ or yolk- sac resorbed larvae. The greatest increase occurred in 18: 3n-3 from 6.4±0.1 to 106.2±13.1 g mg lipid^–1 larva^–1, and the greatest decrease occurred in 22: 6n-3 (181.2±12.4 to 81.4±6.2 g mg lipid^–1 larva^–1). In starved larvae, at the end of 15 days, all the fatty acids, except 18: 0, 20: 3n-3 and 20: 4n-6, decreased significantly (P<0.05) from the levels in day of hatch and/or yolk- sac resorbed larvae.     

Investigation of highly unsaturated fatty acid metabolism in the Asian sea bass, <Emphasis Type="Italic">Lates calcarifer</Emphasis>

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 toward 18:3n-3 and 18:2n-6, and Elovl5 exhibited elongation activity for C_18–20 and C_20–22 elongation and a trace of C_22–24 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.     

Dynamics of total lipids and fatty acids during embryogenesis and larval development of Eurasian perch (Perca fluviatilis)

Total lipid and fatty acid compositions were determined during embryogenesis and larval development in Eurasian perch (Perca fluviatilis). During embryonic development, perch did not catabolize lipids and fatty acids as an energy source. However, during larval development, there was an exponential relationship between the decrease in total lipids and the duration of starving (r ²=0.9957) and feeding (r ²=1). The duration of the starving period (10 days post hatching) was shorter than the feeding period (35 days post hatching). In both starved and fed larvae, there is an apparent preference in utilization of polyunsaturated fatty acids followed by monounsaturated fatty acids. Saturated fatty acids were utilized by neither fed perch larvae nor by starved perch larvae. In starved larvae, palmitoleic 16:1(n-7) and oleic 18:1(n-9) acids were the preferentially monounsaturated fatty acids catabolized and their contribution as energy source from total fatty acids catabolized over the first week was 37.6%. In fed larvae, these 2 nutrients were also the most monounsaturated fatty acids utilized as energy source and possibly also as precursors for others monounsaturated fatty acids biosynthesis. During the same period and among (n-6) polyunsaturated fatty acids, starved perch utilized less linoleic 18:2(n-6) and arachidonic 20:4(n-6) acids than fed larvae despite the fact that the starved perch were in more unfavorable nutritional conditions. In the case of (n-3) fatty acids, starved larvae utilized more linolenic acid 18:3(n-3) and less eicosapentaenoic 20:5(n-3) acid and docosahexaenoic 22:6(n-3) acid than fed perch. Starved larvae probably spared 20:5(n-3) and 22:6(n-3) for physiological functions.     

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).

     

南极磷虾粉脂质及脂肪酸组成分析

为分析南极磷虾粉的脂质成分,实验采用薄层层析法分离制备不同脂质组分,通过酸甲酯化衍生结合气相色谱/质谱法对不同脂质的脂肪酸组成进行比较研究.结果显示:①南极磷虾粉中脂质丰富,总脂含量高达11.37％,且以甘油三酯、磷脂和游离脂肪酸为主,胆固醇含量较低;②总脂脂肪酸主要为C16:0、C18:1n-9、C18:1n-7、C20:5n-3和C22:6n-3,且C20:5n-3和C22:6n-3占总脂肪酸含量的30.67％,表明南极磷虾粉在脂质方面具有较高的营养价值和开发潜力.③不同脂质的脂肪酸组成差异显著,胆固醇酯和甘油三酯中饱和脂肪酸和单不饱和脂肪酸含量均显著高于游离脂肪酸和磷脂(P＜0.05);而游离脂肪酸和磷脂富含多不饱和脂肪酸,两者含量分别为48.50％和49.96％,远高于胆固醇酯和甘油三酯的13.11％和24.36％,具有显著差异(P＜0.05).     

养殖和野生亚东鲑机体成分比较分析

亚东鲑(Salmo trutta fario)是鲑属鱼类在青藏高原仅有的鱼类种群,是亚东地区全国农产品地理标志产品。本研究对西藏亚东地区野生和养殖亚东鲑的常规营养成分及各组织氨基酸和脂肪酸组成进行分析,旨在比较野生和养殖亚东鲑营养成分的异同,为养殖亚东鲑的品质评价和饲料配方的完善提供参考信息。实验采集亚东河中野生亚东鲑和亚东渔业产业园中使用配合饲料养殖的亚东鲑各10尾用于相关成分分析,每尾为一个独立样本。结果显示,野生组肥满度显著低于养殖组,而全鱼水分和灰分显著高于养殖组。全鱼粗蛋白、粗脂肪以及肝脏常规成分在野生组和养殖组间无显著差异。野生组肌肉粗脂肪显著低于养殖组,而水分含量显著高于养殖组。野生组全鱼必需氨基酸总量显著高于养殖组,且野生组肌肉中苏氨酸、缬氨酸、苯丙氨酸、赖氨酸和甘氨酸含量显著高于养殖组。野生组全鱼、肌肉和肝脏中饱和脂肪酸、n-3多不饱和脂肪酸(n-3 PUFA)总量、EPA、C20:4n-6以及肌肉中DHA含量高于养殖组,而全鱼和组织中单不饱和脂肪酸和n-6 PUFA总量低于养殖组。综上所述,目前养殖亚东鲑和野生亚东鲑在机体成分上存在较大差异。由于养殖鱼类体成分很大程度上反映了饲料组成,因此,亚东鲑养殖中饲料营养组成可能有待进一步优化。     

Essential fatty acid deficiency in freshwater fish: the effects of linoleic, α-linolenic, γ-linolenic and stearidonic acids on the metabolism of [1-14C]18:3n-3 in a carp cell culture model

The desaturation of [1-¹⁴C]18:3n-3 to 20:5n-3 and 22:6n-3 is enhanced in an essential fatty acid deficient cell line (EPC-EFAD) in comparison with the parent cell line (EPC) from carp. In the present study, the effects of competing, unlabeled C₁₈ polyunsaturated fatty acids (PUFA), linoleic (18:2n-6), -linolenic (18:3n-3), -linolenic (18:3n-6) and stearidonic (18:4n-3) acids, on the metabolism of [1-¹⁴C]18:3n-3 were investigated in EPC-EFAD cells in comparison with EPC cells. The incorporation of [1-¹⁴C]18:3n-3 in both cell lines was significantly reduced by competing C₁₈ PUFA, with the rank order being 18:4n-3>18:3n-3 = 18:2n-6>18:3n-6. In the absence of competing PUFA, radioactivity from [1-¹⁴C]18:3n-3 in EPC cells was predominantly recovered in phosphatidylethanolamine followed by phosphatidylcholine. This pattern was unaffected by competing n-6PUFA, but n-3PUFA reversed this pattern as did essential fatty acid deficiency in the presence of all competing PUFA. The altered lipid class distribution was most pronounced in cells supplemented with 18:4n-3. Competing C₁₈ PUFA significantly decreased the proportions of radioactivity recovered in 22:6n-3, pentaene and tetraene products, with the proportions of radioactivity recovered in 18:3n-3 and 20:3n-3 increased, in both cell lines. However, the inhibitory effect of competing C₁₈ PUFA on the desaturation of [1-¹⁴C]18:3n-3 was significantly greater in EPC-EFAD cells. The magnitude of the inhibitory effects of C₁₈ PUFA on [1-¹⁴C]18:3n-3 desaturation was dependent upon the specific fatty acid with the rank order being 18:4n-3>18:3n-3>18:2n-6, with 18:3n-6 having little inhibitory effect on the metabolism of [1-¹⁴C]18:3n-3 in EPC cells. The differential effects of the C₁₈ PUFA on [1-¹⁴C]18:3n-3 metabolism were consistent with mass competition in combination with increased desaturation activity in EPC-EFAD cells and the known substrate fatty acid specificities of desaturase enzymes. However, the mechanism underpinning the greater efficacy with which the unlabeled C₁₈PUFA competed with [1-¹⁴C]18:3n-3 in the desaturation pathway in EPC-EFAD cells was unclear.