Influence of dietary polar lipids' quantity and quality on ingestion and assimilation of labelled fatty acids by larval gilthead seabream

Dietary supplementation of phospholipids seems to be extremely important to promote growth and survival in fish larvae. Several studies also suggest the importance of n-3 highly unsaturated fatty acids (HUFA) rich phospholipids to further enhance larval performance. In the present study, four different diets were formulated in order to compare the effect of total dietary polar lipid contents, of soya bean lecithin supplementation and of feeding n-3 HUFA in the form of neutral or polar lipids on ingestion and incorporation of labelled fatty acids in gilthead seabream larvae. These diets were prepared including radiolabelled fatty acids from palmitoyl phosphatidylcholine, glycerol trioleate, free oleic acid (FOA) and free eicosapentaenoic acid (FEPA) and were fed to 25 day-old larvae. The results of these experiments showed that the elevation of the dietary polar lipid levels significantly improved microdiet ingestion, regardless of the origins of the polar lipids. This effect caused an improved incorporation of phosphatidylcholine fatty acids to the larval polar and total lipids (TL) as the dietary polar lipids increased. Nevertheless, a better incorporation of fatty acids from dietary polar lipids in comparison with that of fatty acids from dietary triglycerides into larval lipids was found in gilthead seabream, whereas a better utilization of dietary triglycerides fatty acids than dietary free fatty acids could also be observed. Besides, the presence of n-3 HUFA rich neutral lipids (NL) significanlty increased the absorption efficiency of labelled oleic acid from dietary triglycerides, but the presence of n-3 HUFA rich polar lipids, particularly improved the incorporation of FEPA. This fatty acid was preferentially incorporated into larval polar lipids in comparison with FOA.     

Effect of a dietary phospholipid supplementation on growth and fatty acid composition of European sea bass (Dicentrarchus labrax L.) and turbot (Scophthalmus maximus L.) juveniles from weaning onwards

Two 40-day feeding trials using extruded diets were conducted to assess the effect of a dietary phospholipid (PL) supplementation on growth, survival and fatty acid composition of European sea bass (Dicentrarchus labrax) and turbot (Scophthalmus maximus) from weaning onwards. Two dietary treatments (FO and PL) were tested; both had an identical extruded basis (92.5% total diet weight) coated with a different lipid fraction (7.5% total diet weight). Diet PL contained 2% egg yolk PL (69% pure). In diet FO the PL was replaced by hydrogenated coconut oil. The isolipidic diets contained an equal amount of fish oil ethyl esters providing 1.6% (% diet dry weight) of n-3 highly unsaturated fatty acids (HUFA). A diet water stability test showed no effect of the PL supplementation on the leaching of the dietary fatty acids. In both fish species weight, but not survival, significantly increased as a result of PL supplementation. Weaning onto the experimental diets resulted in similar changes in the relative percent levels of fatty acids in both species. In general, the percentage of saturated fatty acids levelled off after a rapid increase, while monoenes increased after an initial decrease. Total n-3 polyunsaturated fatty acids (PUFA) decreased and total n-6 PUFA remained almost constant. The major effect of the dietary PL on fish fatty acid composition was a 50% increase in n-6 and n-3 HUFAs compared to the PL-free FO diet. The rise in n-6 HUFA may have reflected the higher moiety in the dietary PL. On the other hand this was not the case for the n-3 HUFA since they represented only low levels in the PL fraction (0.1%) compared to that provided by the ethyl esters (1.6%) suggesting a more efficient incorporation of the PL n-3 HUFA than of the ethyl ester n-3 HUFA. A second hypothesis is that the dietary PL may have favored the incorporation of the dietary ethyl ester n-3 HUFA.     

Dietary effects of n-3 highly unsaturated fatty acid levels on egg and larval quality, and the fatty acid composition of the eggs of Chilean flounder Paralichthys adspersus broodstock

The effects of dietary n-3 highly unsaturated fatty acid (n-3 HUFA) on eggs and larval quality were investigated in the Chilean flounder Paralichthys adspersus . Broodstock were fed with three formulated diets with similar proximate compositions but different n-3 HUFA (2.1%, 3.1% or 4.1%) estimated levels from 5 months before and during the spawning period. The diet with an intermediate n-3 HUFA level resulted in a significantly higher ( P <0.05) percentage of buoyant eggs (68.2 ± 2.9%), fertilization (92.8 ± 3.9%), normal cell cleavages (93.5 ± 1.9%), hatching rate (87.7 ± 4.1%) and normal larvae (76.3 ± 3.7%) compared with the other two diets. In contrast, high levels of n-3 HUFA produced larvae with a higher survival capacity when subjected to fasting. The diet with the lowest content of n-3 HUFA produces lower quality eggs and larvae. The n-3 HUFA level in eggs increased with an increase in the dietary level, and the n-3/n-6 ratios were 1:1, 2:1 and 3:1. The DHA/EPA and EPA/ARA ratios of 2 and 4 in eggs, respectively, were associated with improved egg and larval quality and were similar to the ratios found in eggs from wild broodstock. Attainment of optimal fatty acid contents in broodstock diets is one of the key factors for producing the high-quality spawning required for managed culture of this flounder.     

Effects of dietary levels and ratio of phosphatidylcholine and phosphatidylinositol on the growth,survival and deformity levels of Atlantic cod larvae and early juveniles

Effects of two weaning diets that differed in phospholipid (PL) classes on growth, survival and deformities of cod larvae and early juveniles were evaluated. Cod larvae were fed rotifers until 21 days post hatch (dph) and then weaning onto dry diet started. One group of larvae were fed a control diet with low levels of phosphatidylcholine (PC), PE and phosphatidylinositol (PI) and the other group of larvae were fed with an experimental diet containing higher levels of PC, PE and PI. Larvae fed with the control diet were significantly smaller than larvae fed with the experimental diet at the end of the experiment. Swim bladder abnormalities were significantly higher in larvae fed with control diet at 35 dph than the larvae fed with experimental diet; however, no significant difference was evident at 42 dph. Vertebral deformities were significantly higher in larvae fed with control diet and scoliosis was significantly different between the treatments. Survival was also significantly higher in the experimental group. Our results indicate that dietary levels of PL, PC and PI may affect the cod larval growth, survival and deformities. More detail studies are needed to find out the optimal levels of these important PL classes in larval cod diets.     

Incorporation of fatty acids from dietary neutral lipid in eye, brain and muscle of postlarval turbot fed diets with different types of phosphatidylcholine

Previous results demonstrated the stimulating effect of soybean phosphatidylcholine (PC) on the utilization of dietary neutral lipid in larval and postlarval fish. The present study further investigated the effect of the degree of saturation of dietary PC on the enhancement of dietary fatty acid incorporation in lipids of turbot. Newly-weaned turbot were fed for 20 days on four isolipidic diets containing the same amount of highly unsaturated fatty acids (HUFA), presented either as neutral lipid, i.e. fish oil ethyl esters, or as polar lipid. Diet FO was a phospholipid-free control diet. Diets HPC, SPC and FPC were supplemented with 3% hydrogenated soybean PC, 3% native soybean PC and 3% marine fish roe PC, respectively.The three PC-supplemented diets resulted in better growth and higher muscle triacylglycerol levels than the PC-free diet FO. The fish fatty acids were determined in 3 lipid classes (neutral lipid, PC, phosphatidylethanolamine) of 3 organs or tissues (eye, brain and muscle). Despite the identical amounts of n-6 and n-3 fatty acids provided by the soybean oil and by the HUFA ethyl esters, the substitution of 3% hydrogenated coconut oil in diet FO by 3% hydrogenated PC in diet HPC caused, averaged over the various tissues and lipid classes, a 7 to 12% higher incorporation of 18:2n-6, 20:4n-6, 20:5n-3 and a 32% higher 22:6n-3 level in turbot lipid. Diet HPC appeared as efficient as diet SPC for enhancing the incorporation of the n-3 HUFA from the ethyl esters. Feeding diet FPC, in which the n-3 HUFA were provided through the marine PC source, resulted in slightly higher levels of these fatty acids in the fish than feeding the ethyl ester HUFA diets, even if supplemented with PC. Present results confirm the positive effect of PC, either hydrogenated or native, on the utilization of fatty acids provided in the diet as neutral lipid. The slightly higher incorporation of HUFA, when esterified on dietary PC instead of neutral lipid, raises the question regarding the form of intestinal absorption of PL in fish.p>     

Biomarkers of bone development and oxidative stress in gilthead sea bream larvae fed microdiets with several levels of polar lipids and α‐tocopherol

Although dietary marine phospholipids are able to improve culture performance of marine fish larvae in a further extend than soybean lecithin, both types of phospholipids (PL) markedly increase oxidative risk. The inclusion of a fat‐soluble antioxidant such as the vitamin E α‐tocopherol could allow a better control of oxidative stress. The objective of this study was to determine the combined effect of graded levels of α‐tocopherol with different levels and sources of krill phospholipids (KPL) and soybean lecithin (SBL) on growth, survival, resistance to stress, oxidative status, bone metabolism‐related genes expression and biochemical composition of sea bream larvae. Sea bream larvae were completely weaned at 16 dph and fed for 30 days seven microdiets with three different levels of PL (0, 40 and 80 g kg^?1 diet) and two of α‐tocopherol 1500 and 3000 mg kg^?1 diet. Sea bream larvae fed diets without PL supplementation showed the lowest survival, growth and stress resistance, whereas increase in PL, particularly KPL, markedly promoted larval survival and growth. However, feeding SBL markedly increased TBARs and GPX gene expression increasing the peroxidation risk in the larvae. Besides, KPL inclusion improved incorporation of n‐3 HUFA and, particularly, EPA into larval tissues, these fatty acids being positively correlated with the expression of BMP‐4, RUNX 2, ALP, OC and OP genes and to bone mineralization for a given larval size class. The increase in dietary α‐tocopherol tends to improve growth in relation to the n‐3 HUFA levels in the diet, denoting the protective role of this vitamin against oxidation. Indeed, dietary α‐tocopherol decreased the oxidative stress in the larvae as denoted by the reduction in larval TBARs contents and gene expression of SOD and CAT, but not GPX. Thus, increase in dietary α‐tocopherol effectively prevented the formation of free radicals from HUFA, particularly EPA, but did not affect the incidence of bone anomalies or the expression of genes related to osteogenetic processes.     

Body lipid and fatty acid composition in male gilthead seabream broodstock at different stages of the reproductive cycle: effects of a diet lacking n-3 and n-6 HUFA

Total lipid (TL), lipid classes and fatty acid composition of neutral (NL) and polar (PL) lipids were studied in the gonads, liver and muscle of gilthead seabream males ( Sparus aurata ) fed a control diet (diet C) or an n-3 and n-6 highly unsaturated fatty acids (HUFA)-deficient diet (diet D), at different stages of the reproductive cycle. Between pre-spermatogenesis (November) and spermatogenesis (March), the lipid content was high and particularly rich in cholesterol, phosphatidylcholine and phosphatidylethanolamine in gonads from both dietary groups. At post-spermatogenesis (June), TL and especially PL dramatically decreased in the gonads from both groups. However, at this period diet C fish gonads were richer in triacylglycerides (TAG) than those from diet D fish. The liver lipid contents and particularly TAG were over 200% lower in June than in March for both groups. Nevertheless, the most noteworthy depletion of lipids during this period was achieved by the n-3 HUFA in diet D fish. Conversely, arachidonic acid (20:4n-6) did not decrease in NL or PL from gonads and liver in groups C and D. Muscle lipids from diet C fish were relatively insensitive to seasonal influences. However, in June, the muscle TAG content was significantly reduced in diet D fish.     

The importance of dietary HUFA for meagre larvae (Argyrosomus regius; Asso, 1801) and its relation with antioxidant vitamins E and C

Despite the interest of meagre (Argyrosomus regius) as a fast‐growing candidate for Mediterranean aquaculture diversification, there is a lack of information on nutrition along larval development. Importance of highly unsaturated fatty acids (HUFA) and the antioxidant vitamins E and vitamin C has not been investigated yet in this species. Six diets with two levels of HUFA (0.4% and 3% dw), two of vitamin E (1500 and 3000 mg kg^?1) and two of vitamin C (1800 and 3600 mg kg^?1) were fed to 15 dah meagre larvae. Larval growth in total length and dry body weight was significantly lowest in larvae fed diet 0.4/150/180 and showed few lipid droplets in enterocytes and hepatocytes and lower HUFA contents than the initial larvae. Increase in dietary HUFA up to 3%, significantly improved larval growth and lipid absorption and deposition. Besides, among fish fed 3% HUFA, increase in vitamin E and vitamin C significantly improved body weight, as well as total lipid, 22:6n‐3 and n‐3 fatty acids contents in the larvae. Thus, the results showed that 0.4% dietary HUFA is not enough to cover the essential fatty acid requirements of larval meagre and a high HUFA requirement in weaning diets is foreseen for this species. Besides, the results also pointed out the importance of dietary vitamin E and C to protect these essential fatty acids from oxidation, increase their contents in the larvae and promote growth, suggesting high vitamin E and C requirements in meagre larvae (higher than 1500 and 1800 mg kg^?1 for vitamin E and vitamin C respectively).     

Effects of dietary phospholipid and highly unsaturated fatty acid on the gonadal development, tissue proximate composition, lipid class and fatty acid composition of precocious Chinese mitten crab, Eriocheir sinensis

Precocious puberty is a major constraint to the economical rearing Eriocheir sinensis farming. Although dietary phospholipid (PL) and highly unsaturated fatty acids (HUFA) reportedly enhanced ovarian development in normal adult E. sinensis , it had opposite effects of reducing precocity in juveniles . This study investigated the effects of dietary PL and HUFA on survival, gonadal development and biochemical composition of precocious E. sinensis . Two diets with PL and HUFA supplementation (diet A) and deficient (diet B) were formulated and fed to precocious E. sinensis . Although no significant differences were found on survival and gonadosomatic index of crabs fed both diets, crabs fed diet B had significantly higher hepatosomatic index and hepatopancrean lipid content than crabs fed diet A. The percentages, 14:0, 18:1n-9, 20:5n-3 and 22:6n-3, in hepatopancreas, muscle and gonads were positively correlated to the diets. Interestingly, the crabs fed diet B accumulated significantly higher 18:2n-6 in gonads and muscle than those fed diet A. Furthermore, substantially higher 20:4n-6 was detected in the gonad and muscle than in the diets. These results suggested that dietary PL and HUFA had limited effects on male gonad biochemical composition. Meanwhile, the precocious E. sinensis may be capable of negating the negative effects of HUFA deficiency by enhanced tissue 18:2n-6 accumulation and converting 18:2n-6 to 20:4n-6.     

Influences of dietary fatty acid profile on growth, body composition and blood chemistry in juvenile fat cod (Hexagrammos otakii Jordan et Starks)

This study was conducted to investigate the influence of dietary lipid source and n‐3 highly unsaturated fatty acids (n‐3 HUFA) level on growth, body composition and blood chemistry of juvenile fat cod. Triplicate groups of fish (13.2 ± 0.54 g) were fed the diets containing different n‐3 HUFA levels (0–30 g kg^?1) adjusted by either lauric acid or different proportions of corn oil, linseed oil and squid liver oil at 100 g kg^?1 of total lipid level. Survival was not affected by dietary fatty acids composition. Weight gain, feed efficiency and protein efficiency ratio (PER) of fish fed the diets containing squid liver oil were significantly (P < 0.05) higher than those fed the diets containing lauric acid, corn oil or linseed oil as the sole lipid source. Weight gain, feed efficiency and PER of fish increased with increasing dietary n‐3 HUFA level up to 12–16 g kg^?1, but the values decreased in fish fed the diet containing 30 g kg^?1 n‐3 HUFA. The result of second‐order polynomial regression showed that the maximum weight gain and feed efficiency could be attained at 17 g kg^?1 n‐3 HUFA. Plasma protein, glucose and cholesterol contents were not affected by dietary fatty acids composition. However, plasma triglyceride content in fish fed the diet containing lauric acid as the sole lipid source was significantly (P < 0.05) lower than that of fish fed the other diets. Lipid content of fish fed the diets containing each of lauric acid or corn oil was lower than that of fish fed the diets containing linseed oil or squid liver oil only. Fatty acid composition of polar and neutral lipid fractions in the whole body of fat cod fed the diets containing various levels of n‐3 HUFA were reflected by dietary fatty acids compositions. The contents of n‐3 HUFA in polar and neutral lipids of fish increased with an increase in dietary n‐3 HUFA level. These results indicate that dietary n‐3 HUFA are essential and the diet containing 12–17 g kg^?1 n‐3 HUFA is optimal for growth and efficient feed utilization of juvenile fat cod, however, excessive n‐3 HUFA supplement may impair the growth of fish.     

The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

Nutrition of Marine Fish Larvae

ABSTRACT

In the hatchery production of aquatic animals for aquaculture, livefoods such as diatoms; rotifer, Brachionus plicatilis and brine shrimp, Artemia salina, have been used throughout the world. However, such production requires large facilities, maintenance expenses, and labor to produce a desired amount of live foods constantly and reliably. Also, the nutritive value of planktonic organisms is occasionally variable, indicating that the dietary quality of these live foods varies with the content of n-3 highly unsaturated fatty acid (n-3 HUFA). Therefore it is necessary to develop microparticulate diets as a substitute for live foods to further increase theproductivity of seed for fish culture. The nutritional components of microparticulate diets for fish larvae should be determined on the basis of requirements of the larval fish for proteins and amino acids, lipids and fatty acids, carbohydrates, vitamins, and minerals. Moreover, the efficient development of microparticulate diets for the fish larvae has promoted the improvement of nutritional requirement studies. The present reviewconcerning the nutrition of marine fish larvae focuses on the proteins, amino acids, peptides, fatty acids, phospholipids, depigmentation of flatfish, stress tolerance of lipids, incorporation of HUFA in neural tissues, HUFA in egg and larvae, HUFA enrichment of live food, carbohydrates, vitamins, energy source during embryo and larval stages, enzyme supplement in microparticulate diets, and application of microparticulate diets inaquaculture.     

Effect of krill phospholipids versus soybean lecithin in microdiets for gilthead seabream (Sparus aurata) larvae on molecular markers of antioxidative metabolism and bone development

The objective of the present study was to compare the effectiveness of dietary marine phospholipids (MPL) obtained from krill and soybean lecithin (SBL) on the rearing performance and development of seabream (Sparus aurata) larvae. Larvae were fed from 16 to 44 day posthatching (dph) five formulated microdiets with three different levels (50, 70 and 90 g kg^–1) of phospholipids (PL) obtained either from an MPL or from a SBL source. Larvae‐fed MPL show a higher survival, stress resistance and growth than those‐fed SBL, regardless the dietary PL level. Overall, the increase in MPL up to 70 g kg^–1 total PL in diet was enough to improve larval gilthead seabream performance, whereas even the highest SBL inclusion level (90 g kg^–1 PL) was not able to provide a similar success in larval growth or survival. Inclusion of SBL markedly increased the peroxidation risk as denoted by the higher TBARs in larvae, as well as a higher expression of CAT, GPX and SOD genes. Moreover, SBL tends to produce larvae with a lower number of mineralized vertebrae and a lower expression of osteocalcin, osteopontin and BMP4 genes. Finally, increasing dietary MPL or SBL lead to a better assimilation of polyunsaturated fatty acids in the larvae, n‐3HUFA (especially 20:5n‐3) or n‐6 fatty acids (especially 18:2n‐6), respectively. In conclusion, MPL had a higher effectiveness in promoting survival, growth and skeletal mineralization of gilthead seabream larvae in comparison with SBL.     

Essentiality of Dietary n-3 Highly Unsaturated Fatty Acids in Juvenile Japanese Flounder Paralichthys olivaceus

This study was conducted to confirm the essentiality of dietary n-3 highly unsaturated fatty acids (n-3 HUFA) and to investigate the effects of dietary lipid sources on growth performance, liver, and blood chemistry in juvenile Japanese flounder. Three replicate groups of fish (average weighing 3.0 g) were fed experimental diets containing lauric acid ethyl ester, soybean oil, soybean and linseed oils mixture, and squid liver oil as lipid sources for 13 wk. No significant difference was observed in survival among all groups ( P >0.05). Weight gain, feed efficiency and protein efficiency ratio of fish fed the squid liver oil diet containing high n-3 HUFA level were significantly higher than those of fish fed the other diets ( P 0.05). Saturated and monounsaturated fatty acids of liver polar and neutral lipid fractions in fish fed the diet containing lauric acid tended to increase compared to those of the other groups. Fish fed the diets containing soybean and/or linseed oils, which contained high contents of 18:2n-6 and 18:3n-3, respectively, showed the highest contents of 18:2n-6 and 18:3n-3 in both lipid fractions of the liver ( P 0.05). Significantly higher content of n-3 HUFA was observed in both lipid fractions of the liver from fish fed the diet containing squid liver oil than for fish fed the other diets ( P 0.05). Total cholesterol, glucose, and glutamic-oxaloacetic acid transaminase in plasma were significantly affected by dietary lipids ( P 0.05). Histologically, the liver of fish fed the diet containing squid liver oil had a clear distinction between nuclear and cytoplasm membranes; however, cytoplasm of fish fed the diets containing lauric acid and soybean oil was shrunken, and the hepatic cell outline was indistinguishable. It is concluded that the dietary n-3 HUFA is essential for normal growth, and that the dietary lipid sources affect growth performance, liver cell property, and blood chemistry in juvenile Japanese flounder.     

Early ontogeny-related changes of the fatty acid composition in the Percichthyid fishes trout cod,Maccullochella macquariensis and Murray cod,M. peelii peelii

Changes in the fatty acid profiles of the Percichthyid fish trout cod, Maccullochella macquariensis (Cuvier), and Murray cod, M. peelii peelii (Mitchell), two Australian native freshwater fish species, were investigated during early development from egg to yolk-sac-resorbed larval stage. In the two Percichthyid fishes polyunsaturated fatty acids (PUFA) accounted for more than 50 % of the 19 quantified fatty acids in total lipid. The fatty acids that occurred in the highest abundance in both trout cod and Murray cod, in all developmental stages, in order, were docosahexaenoic acid [DHA 22:6(n-3)], arachidonic acid [AA 20:4(n-6)], oleic acid [18:1(n-9)] and palmitic acid (16:0), all of which exceeded 100 μg per mg total lipid in most instances. The ratio of 22:6(n-3) to eicosapentaenoic acid- 20:5(n-3) in eggs of trout cod and Murray cod was 5.4:1 and 7.3:1, respectively, and remained almost unchanged through development, and was considerably higher than the 2:1 ratio generally reported for fish eggs. In trout cod, 11 of the 19 fatty acids in total lipid decreased during the transformation from egg to yolk-sac-resorbed larva. In Murray cod, only 16:1(n-7) showed a significant decrease whilst 20:4(n-6) increased significantly with development. Overall, there was a tendency in both species to conserve n-3 and n-6 series highly unsaturated fatty acids (HUFA), suggesting their essentiality in first feeding larvae. These observations are discussed in relation to the feeding habits of trout cod and Murray cod, which are top order, freshwater carnivores.     

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.     

Artemia enriched with high n-3 HUFA may give a large improvement in performance of Atlantic halibut (Hippoglossus hippoglossus L.) larvae

Atlantic halibut larvae were fed Artemia enriched with two different oil emulsions (cod liver oil and 2050TG) from first feeding to 70 days after first-feeding (dpff). Larvae fed 2050TG enriched Artemia had better growth, survival and eye migration than larvae fed the cod liver oil enriched Artemia, while pigmentation rate was similar in the two groups. In addition to the difference in fatty acids, the two emulsions differed in lipid class composition, since 2050TG is a synthetic oil and a mixture of mono-, di- and tri-acylglycerol, while cod liver oil is a tri-acylglycerol. Total lipid level, estimated as fatty acid methyl esters (FAME) was similar in the two Artemia types, but sum of n-6 and n-3 fatty acids, arachidonic acid (20:4n-6, ARA), docosahexaenoic acid (22:6n-3, DHA) and eicosapentaenoic acid (20:5n-3, EPA) were higher in Artemia enriched with 2050TG than in the cod liver oil enriched Artemia. However, the main difference in fatty acid composition in the larvae, was a higher DHA (% of total fatty acids) in 2050TG larvae than in cod liver oil larvae. The lipid level measured as FAME was up to four times higher in the 2050TG larvae than in the cod liver oil larvae, and the reason for this may have been a better bioavailability of the partly digested lipid in the 2050TG emulsion. The correlation between a high level of lipid in the larval tissues (e.g. high energy status) and improved eye migration in larvae fed the 2050TG enriched Artemia supports the hypothesis that energy limitation on the larval stage may be a cause of the impaired eye migration commonly observed in farmed Atlantic halibut juveniles.     

Effects of a diet lacking HUFA on lipid and fatty acid content of intestine and gills of male gilthead seabream (Sparus aurata L.) broodstock at different stages of the reproductive cycle

A feeding experiment was carried out to determine the effect of a diet lacking n-3 and n-6 highly unsaturated fatty acids (HUFA) on lipid and fatty acid content in intestine and gills of male gilthead seabream (Sparus aurata L.) broodstock at different stages of the reproductive cycle: November (pre-spermatogenesis), March (spermatogenesis), and June (post-spermatogenesis). For this purpose, gilthead seabream broodstock were fed either a control diet (C) or an n-3 and n-6 HUFA-deficient diet (D). The results showed no changes in fatty acid content of polar lipids of intestine and gills from fish fed diet C at different stages of the reproductive cycle. However, significant changes were observed in the fatty acid content of neutral lipids in intestine but not in gills in this group. Thus, between November and March, saturates and n-3 HUFA decreased while monoenes increased. In June, the contents of these fatty acids had returned to their initial values (November). Moreover, in fish fed diet D, the fatty acid content of neutral lipid changed in both intestine and gills. In intestine NL, a decrease in saturates and n-3 HUFA and an increment in monoenes were observed from November to June. In gills, a decrease was also observed in n-3 HUFA from NL along the cycle. Nevertheless, n-6 HUFA content remained unchanged. These results show both tissue specificity in seasonal mobilization of fatty acids linked to reproductive processes and the influence of dietary fatty acids on body composition.     

Beneficial effects of n-3HUFA enriched Artemia as food for larval palmetto bass (Morone saxatilis × M. chrysops)

The beneficial effects of feeding n−3 highly unsaturated fatty acids (HUFA ≥ 20 carbon fatty acids with three or more double bonds) to palmetto bass (striped bass × white bass) larvae, 4–30 days of age, were studied using Artemia diets enriched with six n-3HUFA levels. Dietary n-3HUFA concentrations were < 0.03% (control diet), 0.33%, 0.63%, 0.87%, 1.26%, and 2.27% of dry-wt Artemia. Larval n-3HUFA contents were reduced at a faster rate with decreasing dietary n-3HUFA intake, and were significantly different by 30 days posthatch (4–20 mg g⁻¹ dry-wt fish). Starved larvae selectively conserved endogenous n-3HUFA reserves, indicating an essential role of n-3HUFA in larval development. Mass mortality in the control and 0.33% n-3HUFA diets occurred at metamorphosis (26–28 days posthatch). At harvest, all fish, except those fed the two highest n-3HUFA diets, suffered from handling stress (shock syndrome) with increasing severity (25 to 100%) at decreasing dietary n-3HUFA intake. Recovery from shock syndrome was 100% at the 0.63% and 0.87% n-3HUFA diets, 63% at the 0.33% n-3HUFA diet and 0% at the control diet. Post-harvest survival was similar among the four highest dietary n-3HUFA groups (64–73%), whereas the two lowest n-3HUFA groups differed significantly (0 and 37%). Growth promotion was evident with increased dietary n-3HUFA intake as fish fed the highest n-3HUFA diet were twice the size of those fed the control diet (99 vs 52 mg wet-wt). Final fish sizes at the three highest n-3HUFA diets were similar. Given similar culture conditions, a minimum dietary n-3HUFA intake of 1.26% of dry-wt Artermia is recommended to avoid handling stress and promote growth in larval palmetto bass.     

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.