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.     

Fatty Acids Profile,Atherogenic, Thrombogenic,and Polyene Lipid Indices in Golden Grey Mullet (Liza aurata) and Gold Band Goatfish (Upeneus moluccensis) from Mediterranean Sea

ABSTRACT

The aim of this work was to study the seasonal variations of the proximate compositions, fatty acids contents, and related lipid indices (atherogenic index [AI], thrombogenic index [TI], and polyene index [PI]) of golden grey mullet (Liza aurata) and gold band goatfish (Upeneus moluccensis) caught from the northeastern Mediterranean Sea. Proximate compositions and fatty acid contents of both fish species showed statistically significant seasonal variations (p < 0.05). The lipid contents were highest in autumn and lowest in winter in golden grey mullet and gold band goatfish. The main saturated and unsaturated fatty acids were palmitic acid (C16:0), stearic acid (C18:0), palmiteoleic acid (C16:1n-7), oleic acid (18:1n-9c), arachidonic acid (C20:4n-6), eicosapentaenoic acid (EPA, C20:5n-3), and docosahexaenoic acid (DHA, C22:6n-3). The total polyunsaturated fatty acids in gold band goatfish did not show any differences among the seasons, whereas that in golden grey mullet was highest in winter. Levels of EPA, DHA, and n-3/n-6 ratios, as well as AI, TI, and PI, were significantly changed among seasons. The results show that golden grey mullet and gold band goatfish, which are among the important fish in Turkey and of commercial value, are a good source for human consumption in terms of their nutritional quality.     

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).     

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     

Effects of temperature and feed composition on essential fatty acid (n-3 and n-6) retention in Atlantic salmon (Salmo salar L.) parr

Retentions of total n-3 and n-6 essential fatty acids (EFAs) were assessed in Atlantic salmon (Salmo salar L.) parr held at 8 °C and 2 °C until they increased in weight from ca. 19 g to 38 g. Feeds contained sandeel oil or a rapeseed:linseed oil blend at 21 and 34% dietary fat. EFA retention efficiencies [(g EFA gained g EFA ingested-1) × 100] were estimated by the 'mass balance method' from measurements of feed intake, changes in biomass for each tank of fish, and fatty acid compositions of the feeds and fish. The n-3 EFA retentions were higher (overall mean 71%) across feed treatments and temperatures than the n-6 EFA retentions (overall mean 63%). Retentions of the n-3 fatty acids were higher in the fish given the feeds with the lower fat content (77% vs. 65%), implying improved retention with reduced n-3 EFA availability. n-3 EFA retention tended to be higher at 2 °C than at 8 °C, although this was not consistent across feeds. At low temperature there was very high retention of the n-3 EFAs in feeds containing sandeel oil (80%). Such high retention may represent an adaptation response to low temperature. Lower n-6 EFA retentions imply that more n-6 fatty acids were metabolized than n-3 EFAs. Feed oil influenced retention of the n-6 fatty acids, retention being lower for the salmon parr given the feeds containing sandeel oil (56% vs. 71%). This could indicate a higher tissue deposition of n-6 fatty acids when they are freely available via the diet. Abbreviations: AA – arachidonic acid (C20:4 n-6); DHA – docosahexaenoic acid (C22:6 n-3); EFA – essential fatty acid; EPA – eicosapentaenoic acid (C20:5 n-3); HUFA – highly-unsaturated fatty acid (\ge4 double bonds); MUFA – monounsaturated fatty acid (1 double bond); PL – phospholipid; PUFA – poly-unsaturated fatty acids (\ge2 double bonds); SFA – saturated fatty acid (no double bond); TAG – triacylglycerol. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Variation in the Fatty Acid Composition Between and Within Two Mantis Shrimp Species,Harpiosquilla harpax and Miyakea nepa: Impact of Season and Sex

ABSTRACT

Harpiosquilla harpax and Miyakea nepa are two out of 400 marine crustaceans commonly known as mantis shrimp. Mantis shrimp are commercially valuable and can be found in fish markets of some Mediterranean and many Asian countries. This study focused on a comparative analysis of variation throughout the year in regards to proximate and total fatty acid composition in the muscle of male and female H. harpax and M. nepa. The most abundant fatty acids identified in the muscle of both species were C16:0, C18:0, docosahexaenoic acid (DHA; C22:6n-3), eicosapentaenoic acid (EPA; C20:5n-3), and C20:4n-6. Results of this study revealed that the muscle fatty acid profiles of H. harpax and M. nepa were not significantly different (p > 0.05), except for C14:0, C16:0, C22:0, and DHA amounts, and were mostly influenced by sex within each sampling month. Generally, the muscle of H. harpax contained higher (p < 0.05) levels of DHA than the muscle of M. nepa. The male mantis shrimp, within each species and most of the sampling months, appeared to have higher (p < 0.05) DHA, total n-3 polyunsaturated fatty acid (PUFA), and PUFA/saturated fatty acid ratio compared to the females. Both species could be considered as a good source of n-3 PUFA in the human diet.     

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。     

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.     

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.     

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 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.     

Biochemical composition of juvenile cultured vs. wild silver pomfret, <Emphasis Type="Italic">Pampus argenteus</Emphasis>: determining the diet for cultured fish

We evaluated the biochemical composition of juvenile cultured and wild silver pomfret Pampus argenteus. Cultured silver pomfret (SF) had significantly (P < 0.05) lower crude protein content and higher crude lipid content than wild SF. Concentrations of almost all amino acids, except glycine and arginine, were significantly higher (P < 0.05) in wild SF compared to cultured SF. Further, wild SF had a significantly (P < 0.05) higher concentration of total amino acids (TAA), essential amino acids (EAA), and a higher essential amino acid index (EAAI) than cultured SF. Saturated fatty acids (SFA) in cultured SF were significantly higher than in wild SF. However, the content of poly-unsaturated fatty acids (PUFA) and n-3 fatty acids in wild SF was significantly higher than in cultured SF. Mono-unsaturated fatty acids (MUFA) in cultured and wild SF were not significantly different. Fatty acids C16:0 and C18:1 were the principal fatty acids of the SFAs and MUFAs, respectively, in both cultured and wild SF. The results will be useful for determining the dietary requirements for culture of SF.     

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.     

蚕蛹替代鱼粉对吉富罗非鱼肌肉营养成分的影响

试验以300尾初始体重为(44.40±0.41)g的吉富罗非鱼(GIFT,Oreochromis niloticus)为研究对象,分别饲喂用蚕蛹(SP)替代0、25%、50%、75%和100%鱼粉(FM)的5种等氮(32%)等脂(5.5%)的饲料(分别命名SP0、SP25、SP50、SP75和SP100,基础饲料中含8%的鱼粉),每组3重复,每重复20尾鱼,在室内循环水养殖系统进行为期8周的生长试验,旨在探讨蚕蛹替代鱼粉对吉富罗非鱼肌肉营养成分的影响。结果表明:随蚕蛹替代水平上升,吉富罗非鱼肌肉粗蛋白、灰分、总氨基酸、必需氨基酸显著下降,肌肉粗脂肪含量无显著差异;较SP0组,SP100组肌肉总氨基酸和必需氨基酸分别下降3.99%和3.46%。随蚕蛹替代量增加,吉富罗非鱼肌肉C18∶3n-3和C20∶5n-3显著上升,C18∶2n-6显著下降;SP100组吉富罗非鱼肌肉C18∶3n-3和C20∶5n-3较SP0组分别上升105.33%和132.72%。结论:蚕蛹替代鱼粉后显著提高了吉富罗非鱼肌肉C18∶3n-3和C20∶5n-3含量,有利于生产富含n-3PUFA的鱼肉产品。     

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.     

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 salinity on the fatty acid compositions of total lipid and individual glycerophospholipid classes of Atlantic salmon (Salmo salar) and turbot (Scophthalmus maximus) cells in culture

Cells from a relatively stenohaline marine species, turbot (Scophthalmus maximus) (TF) and an anadromous species, Atlantic salmon (AS) were cultured in media supplemented with NaCl to produce OPs varying from 300 to 500 mOsm kg^–1 and the direct effects of OP (salinity) on the fatty acid compositions of the main glycerophospholipid classes were determined. The most dramatic effects of salinity on total lipid fatty acids were observed in polyunsaturated fatty acids (PUFA) in TF cells. There was a graded decrease in the percentage of 18:2n-9, and consequently total n-9 PUFA, and concomitantly increased percentages of both total n-3 and n-6 PUFA with increasing salinity. The increased n-3 and n-6 PUFA was due to significantly increased percentages of the major fatty acids in each of these groups, namely 22:6n-3 and 20:4n-6, respectively. The reciprocal changes in n-9 PUFA and n-3/n-6 PUFA in TF cell total lipid resulted in the percentage of total PUFA not being significantly affected by changes in salinity. The graded decrease in 18:2n-9 with increasing salinity in TF cells was observed in all the major glycerophospholipids but especially PE, PI and PS. Increasing salinity resulted in graded increases in the percentages of 22:6n-3 in PE and PS in TF cells. The quantitatively greatest increase in the percentage of n-6 PUFA in TF cells occurred with 20:4n-6 in PC, PE and PL. There were less significant changes in the fatty acid compositions of glycerophospholipids in AS cells. However, the proportion of total n-3 + n-6 PUFA in PE varied reciprocally with the proportion of dimethylacetals in response to salinity. Similar reciprocal changes between fatty acids in response to salinity were also evident in the quantitatively more minor glycerophospholipids PS and Pl. In PS, the percentage of 22:6n-3 was significantly lower at 400 mOsm kg^–1 whereas the proportion of total monoenes was significantly higher at that salinity. A similar inverse relationship between total monoenes and 20:4n-6 (and, to a lesser extent total saturates) in response to salinity was noted in PI. The results show that environmental salinity, without whole-body physiological stimuli, has direct effects on the fatty acid composition of major glycerophospholipid classes in fish cells and that these effects differ in cells from different fish speciesAbbreviations ANOVA analysis of variance - BHT butylated hydroxytoluene - BSA bovine serum albumin - DMA dimethylacetals - EMEM Eagle's minimal essential medium - FCS fetal calf serum - GC gas chromatography - HBSS Hank's balanced salt solution (without Ca²⁺ and Mg²⁺) - OP osmotic pressure - PC phosphatidylcholine - PE phosphatidylethanolamine - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - TLC thin-layer chromatography     

Fatty acid profiles of two freshwater fish larvae (gudgeon and perch) reared with Brachionus calyciflorus Pallas (rotifer) and/or dry diet

The efficiency of the rotifer Brachionus calyciflorus Pallas as a nutritional source for rearing larvae was studied in a coldwater cyprinid, the gudgeon Gobio gobio (L.), and in a percid, the perch Perca fluviatilis L., through their composition in fatty acids. Rotifer intake affected the fatty acid profiles of the larvae significantly, with an especially remarkable presence of the linoleic family. In gudgeon fed with rotifers, the polyunsaturated fatty acids (PUFA) reached 10.98% of the dry weight of the sample. This rate was highly influenced by the presence of the acids C18:2n-6 and C22:6n-3 which represented 66% of the total PUFA. In perch fed exclusively with rotifers, the PUFA represented 7.27% of the dry weight. In both cases, the ratio n-3/n–6 decreased by 75% and 73% after 10 days of feeding with B. calyciflorus. This variation was probably due to the exogenous supply in acids of the linoleic family through the rotifers and to the fact that these two species of fish seem to favour the mobilization of the n-3 PUFA such as C22:6n-3 for growth and survival. Moreover, with the utilization of rotifers. the reactions of elongation and desaturation from the C18:2n-6 and C18:1n-9 seemed to be much more important in the larvae. Lastly, the transition from a diet based on rotifers to one made up exclusively of frozen Artemia nauplii led to a significant reduction of fatty acids in fish. It reached 60.2% and 26.5% of the total fatty acids in the gudgeon and perch, respectively, and was observed especially at the level of the PUFA. On the other hand, a slight increase of the ratio n-3/n-6 was pointed out in the perch fed a mixed diet (co-feeding with rotifers and dry food), a phenomenon probably due to the reduction of C18:2n-6 in the larvae.     

Fatty acid and amino acid composition of eggs, muscle and midgut glands of freshwater prawns, Macrobrachium rosenbergii (de Man), raised in fertilized ponds, unfertilized ponds or fed prepared diets

Fatty acid and amino acid profiles were determined in three tissues (eggs, muscle and midgut glands) from freshwater prawns, Macrobrachium rosenbergii (De Man), raised unfed in ponds with no organic fertilization, unfed in ponds with organic fertilization or fed a formulated, pelleted diet. Amino acid profiles of tail muscle and eggs were not treatment-dependent. Comparison of amino acid profiles of the diet with those of tissues of animals fed that diet suggest that dietary levels of arginine, histidine, methionine, and especially lysine may be suboptimal. Fatty acid profiles of the midgut gland, tail muscle, and to a lesser extent, eggs reflected the fatty acid composition of the diet in the fed treatment, the organic fertilizer material in the fertilized treatment and natural food organisms in the unfed treatment. Direct consumption of organic fertilizer particles by the prawns is supported by these data. Selective retention of essential fatty acids appears likely in all three treatments. Comparison of the fatty acid composition of prawn eggs and muscle tissue with that of the pelleted diet indicates that 18: 3(n-3), 20: 5(n-3), and especially 20: 4(n-6) are probably below optimal levels. However, under practical culture conditions, prawn growth may not be significantly reduced because of relatively low dietary requirements and contributions from natural foods in the ponds.