The effects of dietary HUFA level on flame angelfish (Centropyge loriculus) spawning,egg quality and early larval characteristics

This study was conducted to determine the effects of dietary highly unsaturated fatty acids (HUFA) on flame angelfish (Centropyge loriculus) reproduction, and egg and larval quality. In the experiment, formulated diets containing 1.8, 2.9 or 3.6% n‐3 HUFA were fed to flame angelfish broodstock (n = 4) for 5 months. Mean fecundity (daily egg production), egg fertilization rates and embryo viability were used as indicators of egg quality. In addition, mean egg diameter, oil globule diameter, per cent hatch, larval size at hatch, per cent survival to yolk exhaustion and larval size at yolk exhaustion were recorded for each treatment. Flame angelfish fed the diet containing 3.6% n‐3 HUFA exhibited significantly increased fecundity, fertilization rates and embryo viability than fish that were fed the other two formulated diets. Egg diameter, egg oil globule diameter, larval size at hatch, larval survival to yolk exhaustion and larval size at yolk exhaustion from the 3.6% n‐3 HUFA broodstock treatment group were not significantly different from those derived from the Control broodstock treatment. These data revealed that flame angelfish egg quality could respond rapidly (within weeks) to maternal dietary changes. Results from this study further support that dietary HUFA composition can significantly affect broodstock reproductive performance as well as subsequent performance of eggs and larvae.     

Utilization of yolk fatty acids by goldfish embryos and larvae

In each of two separate experiments, eggs from a single female goldfish were fertilized, incubated at 22°C and sampled regularly up to day 6 when the larvae could be expected to commence feeding. Hatching normally occurred on Day 4. Lipids were extracted from the eggs and larvae and the neutral lipid and neutral phospholipids were isolated on aminopropyl columns. Fatty acid analysis of the eggs revealed the typical situation in fish where the phospholipids were rich in polyunsaturated fatty acids (PUFA) and the neutral lipids were rich in monounsaturated fatty acids (MUFA). Assay of lipid masses revealed that little depletion of lipid occurred until after hatch and that the neutral phospholipids were the principal fraction consumed. Although the neutral lipid mass did not change substantially during development, its fatty acid profile did. The proportions of several PUFA in the neutral lipids, especially 226(n–3), 205(n–3) and 204(n–6), increased substantially during development while proportions of MUFA and 182(n–6) declined. This appears to be a mechanism by which the larva can retain essential fatty acid released on hydrolysis of phospholipid while deriving the benefits of catabolism of phospholipid as fuel, namely the provision of phosphate and choline for intermediary metabolism and for the synthesis of macromolecules and neurotransmitter.Abbreviations AA arachidonic acid (204(n–6)) - DHA docosahexaenoic acid (226(n–3)) - EPA eicosapentaenoic acid (205(n–3)) - MUFA monounsaturated fatty acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PUFA polyunsaturated fatty acid - SFA saturated fatty acid     

Biochemical composition of eggsin relation to egg quality in the Japanese eel, Anguilla japonica

This paper presents the relationship between egg quality and egg biochemical composition of cultured and wild Japanese eel, Anguilla japonica. Eggs were obtained by artificialinduction of maturation. Fertilization and hatching rates were used as characteristics of egg quality. Egg quality characteristics showed large variation; fertilization rate, 0–96; hatching rate, 0–84%. The biochemical composition also showed a large variation. There was no marked relationship between egg quality and fatty acid contents of eggs, except for n-6 highly unsaturated fatty acids (HUFA). Both the fertilization and hatching ratesincreased proportionally withincreases of the -tocopherol g(-Toc) contentin eggs. A more significant correlation was found between the amount of -Toc relative to the amount of HUFA and egg quality. The results of this study show that the egg quality of Japanese eel is affected by the –Toc level, andin particular, the ratio of -Toc to HUFAin the eggs. Abbreviations: BHT – butylhydroxytoluene; EFA – essential fatty acids; FAME – fatty acid methyl esters; HPLC – high performance liquid chromatography; HUFA – highly unsaturated fatty acids; NADH – nicotinamide adenine dinucleotide; NADPH - nicotinamide adenine dinucleotide phosphate; ROS – reactive oxygen species; -Toc –-tocopherol.     

Changes in lipid class and fatty acid composition during development in pike (Esox lucius L) eggs and larvae

To establish the changes which occur during embryogenesis and early larval development, eggs, yolk sac larvae and swim-up larvae of pike were examined for lipid class and fatty acid composition. At a water temperature of 15.5°C, the embryonic phase was short (6 days) and characterized by a 41.3% decline in the lipid content of eggs, accompanied by large reductions in the amount of phosphatidylcholine (41.4% decrease), sterol esters and triacylglycerols (respectively a 41.2% decrease and a 58.1% decrease), but not phosphatidylethanolamine which increased markedly (35.6%). By the time of yolk sac absorption (7 to 11 days after fertilization) the larvae remained inactive and a limited utilization of lipids was observed. Yolk sac phosphatidylcholine was selectively incorporated into larval bodies while the levels of other lipid classes remained unchanged in the yolk. When the swim bladder was filled and the swimming stage was reached (11 days to 13 days af), the yolk was completely depleted and yolk phosphatidylcholine together with yolk triacylglycerols were catabolised. Yolk phosphatidylethanolamine and yolk sterol esters were partly incorporated into the body lipids. In the subsequent swim-up larval stage (13 to 15 days af), a steady decrease in lipids was observed (41.6%). Fluctuations in the levels of polyunsaturated fatty acids, monounsaturated fatty acids or saturated fatty acids examined from eggs or larvae were consistent with changes in lipid classes during pike development. During yolk sac absorption, pike incorporated yolk PUFA released on hydrolysis of phosphatidylcholine into the larval body. The results are discussed with reference to water temperature and in relation to the ontogenic and ecological context of pike development. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of wolffish eggs at different temperature regimes

Embryonic development of common wolffish (Anarhichas lupus L.) was studied at constant temperatures 5.0, 7.0, 9.0, 11.0, 13.0 and 15.0°C. Duration of development from egg activation to several morphological stages including 50% hatching was determined. At 5.0–11.0°C, the survival rate of eggs to hatching ranged from 51 to 88% with a tendency to increase at 5.0 and 7.0°C. Morphological anomalies, bacterial contamination and large mortalities were observed in eggs incubated at 13.0 and 15.0°C. The period of hatching lasted from 10 to 50 d in different egg groups. Embryo length and yolk sac volume at identical morphological stages of development showed only slight relation to temperature. At lower temperatures newly hatched larvae were longer and at more advanced stages of ontogeny. Normal numbers of fin rays in larvae (mean values 74 for dorsal fin and 46 for anal fin) were observed at 5.0 and 7.0°C and in most larvae at 9.0°C. At 11.0 and 13.0°C, many rays were absent, with mean values for dorsal fin 60 and 39 respectively and for anal fin 28 and 4 respectively. The approximate upper limit for normal development of fin rays appeared to be 9.0 °C.     

Cathepsin L in eggs and larvae of perch Perca fluviatilis: variations with developmental stage and spawning period

This study describes changes in cathepsin L activity during early development in Eurasian perch (Perca fluviatilis) and determines to which extend a precocious proteolysis of the yolk reserves by activated cathepsin L could be involved in the seasonal decline of egg and larval quality. During embryogenesis and early larval stages, samples were collected for determination of fertilization and hatching rates, assays of cathepsin L activity, and evaluation of larval resistance to stress (osmotic shock and fasting). Cathepsin L activity increased significantly during the embryonic development, reaching a maximum of 177.1±12.3 nmol min^–1mg fresh weight^–1 on day 1 post-hatching and significantly decreasing on day 3. Activity increased significantly during the spawning season and individual variations between egg strands became very high during the second half of the spawning period. An inverse relationship was established between the cathepsin L activity in 7-day old eggs and the decrease of hatching rate. A negative exponential regression was calculated between the cathepsin L activity of eggs and the resistance of corresponding newly-hatched larvae to osmotic shock. Resistance to fasting was significantly reduced in larvae from eggs with cathepsin L activity higher or lower than 20 nmol min^–1 mg^–1 (lethal time₅₀=4.7±0.7 and 3.5±0.4 days, respectively). The involvement of cathepsin L in the degradation of yolk reserves and its potential consequences on the quality of perch eggs and larvae are discussed in relation to results reported in salmonid fishes.     

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

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

The essential fatty acid requirement of milkfish (Chanos chanos Forsskal)

The essential fatty acid (EFA) requirement of milkfish was examined by a 12-week feeding trial using defined, purified diets at water temperature of 28–29°C and salinity of 32. The test diets contained varying levels of 18:0 (triglyceride form, TG), 18:3(n–3), 18:2(n–6) and (n–3) highly unsaturated fatty acids (n–3 HUFA). Milkfish juveniles were starved for 7 days and were than fed lipid-free diet for 30 days before the initiation of feeding trials. Low growth and feed efficiency together with high mortalities were observed in fish fed the lipid-free diet as well as in the EFA-deficient diet. Supplementation of 2% 18:2(n–6) to the tristearin based diet did not improve growth rate of milkfish as effectively as feeding with (n–3) fatty acids. The highest weight gain was obtained in milkfish fed a combination of 5% 18:0 + 1.0% 18:3(n–3) + 0.5% 20:5(n–3) + 0.5% 22:6(n–3) although the supplementation of 2% 18:3(n–3) alone or combination of 0.5% 20:5(n–3) + 0.5% 22:6(n–3) to the tristearin based diets were also effective for improvement of growth. Thus, (n–3) fatty acids, such as 18:3(n–3) and (n–3)HUFA were nutritionally more important than 18:2(n–6) for milkfish. The fatty acid composition of the polar lipids from whole body of milkfish juveniles fed the various test diets were influenced by the composition of the dietary fatty acids.     

Lipids of arctic charr,Salvelinus alpinus (L.) II. Influence of dietary fatty acids on the elongation and desaturation of linoleic and linolenic acid

Arctic charr,Salvelinus alpinus L. were fed five test diets containing 0% or 1% of different polyunsaturated fatty acids (PUFA) for 93 days. The fish were injected intraperitoneally with (1–¹⁴C)–18:2(n–6) or (1–¹⁴C)–18:3(n–3), and the bioconversion to longer chain PUFA studied. The conversion rate in neutral lipids was slow, with most label found as the fatty acid injected, while extensive modification took place prior to or during incorporation into polar lipids. Linolenic acid was preferred over linoleic acid as substrate for elongation and desaturation regardless of diet. In polar lipids, the predominant products of (1–¹⁴C)–18:2(n–6) metabolism were generally 20:3(n–6) and 20:4(n–6), while 18:4(n–3), 20:5(n–3) and 22:6(n–3) were the major products of (1–¹⁴C)–18:3(n–3) metabolism. The lack of radioactivity in 22:5(n–6) suggests that 4 desaturation is specific for (n–3) PUFA. Feeding the PUFA deficient diet reduced the 5 desaturation compared to fish maintained on PUFA supplemented diets. The 6 desaturation was only reduced in fish fed C18 PUFA and injected with (1–¹⁴C)–18:3(n–3). Longer chain C20 and C22 PUFA, particularly those of the (n–3) family, exerted some inhibition on the elongation and desaturation of injected fatty acids compared to those fed C18 PUFA. The incorporation of radiolabelled fatty acids into polar lipids of fish fed a commercial diet was very low, and the desaturation neglectible in both polar and neutral lipids, showing that Arctic charr under culture conditions do not convert short chain PUFA to longer chain metabolites.     

Effect of broodstock diets on total lipids and fatty acid composition of larvae of gilthead sea bream (Sparus aurata L.) during yolksac stage

Total lipid content and total lipid fatty acid compositions were studied in larvae at the yolksac stage of gilthead sea bream from two different broodstocks. The two broodstock diets had the same total lipid content but were different with respect to their fatty acid levels. There were differences in the fatty acid composition of total lipids in the two groups of larvae, reflecting the influence of fatty acid levels in the diets fed to the broodstock. The main fatty acids in both larvae lots during yolksac stage were 16:0, 18:1n-9 and 22:6n-3. The desaturation index Δx = [P + Σ (n − x) − Σ (n − x) diet]/P allowed the determination of the relative importance of the Δ9, Δ6, Δ5 and Δ4 desaturations during the egg and larval yolksac stages. The levels of total lipid fatty acids in eggs and larvae during yolksac period were almost constant and very low desaturation and elongation activities were apparent. A competitive inhibition was observed between the fatty acids of the n-6 series with respect to the ones of the n-3 series in those eggs and larvae from broodstock whose diet contained a high proportion of linoleic acid and its derivatives.     

Biochemical composition and quality of captive-spawned cobia Rachycentron canadum eggs

Interest in the commercial production of cobia Rachycentron canadum continues to rise as additional insight is gained into the hardy and fast growing nature of this species. However, research regarding the biochemical composition of captive-spawned eggs and egg and larval quality remains scarce. Such data is essential as a common bottleneck to production is a steady supply of fingerlings for grow-out. This study quantified the biochemical composition and quality of cobia eggs produced over 2 spawning seasons by broodstock on a traditional ‘trash fish’ diet which is commonly fed to tank spawning cobia. Throughout the study, batch fecundity, proportion of floating eggs and percent hatch averaged > 1 million eggs,  0.8 and 70%, respectively. Batch fecundity was significantly higher during the second spawning season as a result of the increased size of the females which weighed 18/22 kg and 22/26 kg at the beginning of each season. A positive correlation was found between the proportion of floating eggs and hatch rate for both spawning seasons. No correlations were found between egg composition (total lipid (30.0 ± 1.1% dry wt), protein (25.4 ± 2.2% dry wt), carbohydrate (2.4 ± 0.3% dry wt), vitamin E (10.2 ± 0.6 μg/g wet wt) or dry weight (119.1 ± 5.5 μg/egg)) and egg quality (proportion of floating eggs, hatch rate, larval growth and larval survival). Further, no differences in egg composition were noted between seasons or over the course of each season. The fatty acid composition of cobia eggs varied between seasons possibly due to changes in the quality of frozen feed (fish, shrimp, squid) given to the broodstock. The only correlation between the fatty acid profile and egg quality was a decrease in the proportion of floating eggs as the total amount of n-3 highly unsaturated fatty acids increased. No relationship between egg quality and amino acid content was noted with the most prominent amino acids being glutamate, leucine, alanine, proline, lysine and aspartate nor were any differences detected between spawning seasons.     

Effect of water hardness on egg hatchability and larval viability of Clarias gariepinus

Studies were conducted to determine the effect of water hardness onClarias gariepinus egg hatchability and larval viability.The fertilized eggs were incubated at 28 °C and with waterhardnesses ranging from 10–700 mg/l CaCO₃. Themean hatching rate fluctuated between 42.31% at hardness of 10mg/land 64.66% at 2000 mg/l. Abnormalities in the larvae were observedbeyond 200 mg/l and increased with increase in water hardness. Thehighest larval survival of 71.05% was recorded at 60 mg/l waterhardness. Based on statistics performed with analysis of variance (ANOVA) andfurther compared with Duncan's multiple range test (p = 0.05), the resultsimplythat very soft water (0–10 mg/l) and very hard water (300mg/l and above) are not suitable for Clariasegg incubation and larval rearing. A water hardness of 30–60mg/l CaCO₃ is recommended for optimal normal hatching,high viability and maximum larval development of Clariasgariepinus.     

Utilization of lipids by Dentex dentex L. (Osteichthyes, Sparidae) larvae during lecitotrophia and subsequent starvation

Total lipids, lipid classes and their associated fatty acids were measured in developing eggs, yolksac larvae and starving larvae (from fertilized egg to day 9 after hatch) of the common dentex Dentex dentex (L., 1758). The larvae of common dentex during lecitotrophia and subsequent starvation consumed 1.6 g of total lipid per larvae per day. The overall decrease was mainly due to utilization of the major neutral lipids, TAG and SE (0.5 and 0.6 g larvae^–1 day^–1, respectively) which was 3.4-fold greater than that of the the major phosphoglycerides (primarily PC by 0.2 g larvae^–1 day^–1). There was net synthesis/conservation of PE during the first half of the study period before it decreased rapidly (0.2 g larvae^–1 day^–1) during the second half. PUFAs were principally catabolized (468.6 ng larvae^–1 day^–1), primarily 22:6(n-3), 20:5(n-3) and 20:4(n-6) (221.8, 58.5 and 12.1 ng larvae^–1 day^–1, respectively). Saturated and monounsaturated fatty acids were also utilized (227.2 and 256.7 ng larvae^–1 day^–1, respectively), principally 16:0 and 18:1(n-9) that were both consumed at 149.8 and 156.7 ng larvae^–1 day^–1, respectively. The rank order of utilization of fatty acids (ng larvae^–1 day^–1) by D. dentex larvae from total lipids, PC and TAG coincided with the order of abundance of the different fatty acids in the respective lipid fractions. However, in PE, the most abundant fatty acid, DHA, was relatively conserved and 16:0, the second most abundant fatty acid, was catabolized to the greatest extent. D. dentex showed a pattern of lipid metabolism during early development similar to that of marine larval fish from temperate waters whose eggs contain high levels of total lipids, including an oil globule, and which preferentially utilize neutral lipids as the primary energy source.     

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.     

Comparison of the lipid profiles from wild caught eggs and unfed larvae of two scombroid fish: northern bluefin tuna (<Emphasis Type="Italic">Thunnus thynnus</Emphasis> L., 1758) and Atlantic bonito (<Emphasis Type="Italic">Sarda sarda</Emphasis> Bloch, 1793)

Lipids and essential fatty acids are determinants of the reproductive process in marine fish, affecting fecundity, egg quality, hatching performance, pigmentation and larval malformation. We have analyzed and characterized the lipids of eggs and unfed larvae of two wild caught scombroid fish, the Atlantic northern bluefin tuna (Thunnus thynnus) and Atlantic bonito (Sarda sarda). Dry matter and total lipid contents, polar and neutral lipid classes and total lipid fatty acid contents were determined in the eggs of bluefin tuna and eggs and unfed larvae during the development of Atlantic bonito. Bluefin tuna eggs had slightly but significantly more dry mass than bonito eggs but very similar lipid content. However, bluefin tuna eggs presented a higher polar lipid content due to increased proportions of phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI). Bonito eggs and larvae showed increasing dry mass and decreasing lipid content with development. The proportion of polar lipids increased due to increased PE, PS and PI, whereas choline-containing polar lipids (phosphatidylcholine and sphingomyelin) remained relatively constant. Free cholesterol also increased, whereas the levels of other neutral lipids, especially triacylglycerol and steryl ester fractions, decreased, presumably due to utilization for energy to drive development. Bluefin tuna eggs had higher levels of n − 3 and n − 6 highly unsaturated fatty acids due to higher docosahexaenoic and arachidonic acid contents, respectively, than bonito eggs. The results are discussed in relation to the lipid and fatty acid requirements of larval scombroid fish in comparison to those of other larval marine finfish species under culture conditions.     

黄颡鱼早期发育阶段受精卵和鱼体脂肪酸组成变化

研究了黄颡鱼受精卵孵化期间和仔鱼发育阶段脂肪含量和脂肪酸的组成变化规律.采用常规化学分析方法和气相色谱法对黄颡鱼从鱼卵受精开始至仔鱼孵化后未投饵的7 d内的脂肪含量和脂肪酸组成进行测定.结果表明,受精卵在整个孵化期间脂肪含量有下降趋势.受精卵中不饱和脂肪酸含量大于饱和脂肪酸含量.受精卵在整个孵化期间各种脂肪酸含量无明显变化.仔鱼孵化后,鱼体总脂肪含量急剧下降,总脂含量从0日龄的4.57%降低到7日龄的0.75%.仔鱼在饥饿期间鱼体脂肪酸组成发生明显变化,单不饱和脂肪酸含量下降最为明显,尤其是C18:1.仔鱼在饥饿期间,脂肪酸按n-9>n-6>n-3顺序被先后利用,黄颡鱼仔鱼发育阶段主要以单不饱和脂肪酸作为能量代谢基质,而C20:4n6(AA)和C22:6n3(DHA)优先于C20:5n3(EPA)被保存下来.     

Biochemical composition and quality of turbot (Scophthalmus maximus) eggs throughout the reproductive season

The present study investigated the biochemical composition and quality of turbot (Scophthalmus maximus) eggs throughout the reproductive season. Results showed that the fertilization, hatching and egg floating rates were variable throughout the reproductive season, with the highest values recorded during the mid-season. Meanwhile, positive correlations were found between fertilization, hatching rate and floating rate. The composition of turbot eggs, including total lipid, protein, carbohydrate, moisture and dry weight showed no significant differences during the reproductive season. Furthermore, no correlations were found between egg compositions and viability parameters (VPs), including fertilization and hatching rates as well as larval deformity rate. However, egg diameter varied and correlated with fertilization, hatching and egg floating rates. The fatty acid in eggs at mid-season had significantly higher levels of C14:0, C16:0, C16:1n-7, C18:0, C18:1, C20:4n-6, C20:5n-3 and C22:6n-3. Moreover, significant relationships were found between fatty acids and VPs. Eggs of the middle season had significantly higher concentration of isoleucine, leucine, lysine, methionine, valine, alanine, aspartate, glutamate and serine, whereas no significant relationships were found between amino acids and VPs. These observations suggest that the biochemical profile of eggs may be useful in evaluating egg quality and improving broodstock management for turbot.     

Effect of dietary supplementation of phospholipids and highly unsaturated fatty acids on reproductive performance and offspring quality of Chinese mitten crab, Eriocheir sinensis (H. Milne-Edwards), female broodstock

This study was conducted to determine the optimum phospholipids: highly unsaturated fatty acids (PL/HUFA) ratio in maturation diets for female Chinese mitten crab, Eriocheir sinensis broodstock. Five isolipidtic and isonitrogenous diets were formulated to contain different amounts of pork lard, fish oil and soy lecithin with resulting PL/HUFA levels (%dry weight) of 1.4/0.3 (Diet 1), 1.6/2.5 (Diet 2), 2.3/2.0 (Diet 3), 3.8/1.5 (Diet 4) and 5.1/0.8 (Diet 5). Diet 1 lacked supplemental fish oil and soy lecithin and served as the control. Each of the five formulated diets was fed to a total of 150 female crabs for a period of 7 months. All diet treatments were triplicated with 50 crabs stocked in a pond as a replicate. After 6 months, male crabs were introduced into the ponds where females were kept, mating, spawning and egg hatching (in seawater) occurred in the following month. The nutritional value of various formulated diets was assessed based on survival, gonadosomatic index (GSI), egg production per female, fecundity (eggs/g female weight) of the females fed those diets and egg and larval (newly hatched zoea I larvae) quality.Females fed Diet 1 had the lowest GSI, egg production per female and fecundity while no significant difference were found for survival, hepatosomatic index (HSI) and the percentage of female spawned among the diet treatments (P > 0.05). Although there were no significant differences in egg diameter, egg wet or dry weights among all treatments (P > 0.05), the highest and the second highest proportions of essential fatty acids (EFA), i.e. 20:5n-3 (EPA), 22:6n-3 (DHA) and HUFA were found in the eggs produced by the females fed Diet 3 and Diet 2, respectively. Statistical analysis showed that EPA, DHA and HUFA in eggs produced by females fed Diet 3 were significantly higher than those from the other treatments (P < 0.05). Meanwhile, zoea I larvae from crabs fed Diet 3 had significantly larger carapace length than those from the other four treatments (P < 0.05). The zoea I larvae from crabs fed Diet 2 and Diet 3 also showed generally better tolerance to starvation and osmotic shock.In conclusion, our results indicated that diet included levels of PL/HUFA around 1.6/2.5 or 2.3/2.0 (% dry weight) fed female E. sinensis broodstock supported increased fecundity and elevated level of HUFA in egg, which in turn resulted in improved overall quality of newly hatched larvae.     

Effects of Temperature on Growth and Efficiency of Yolk Utilisation in Eggs and Pre-feeding Larval Stages of Atlantic Salmon

Three embryonic stages of Atlantic salmon (Salmo salar L.) were subjected to eight constant incubation temperatures (4, 7, 10, 12, 14, 16, 19 and 22 °C) exceeding the range usually experienced in natural conditions. A change in thermal tolerance during the embryonic and larval development was registered: pre-hatching stages showed an upper thermal limit at about 16 °C, while hatched larvae survived until 22 °C. Temperature significantly affects developmental rate, resulting in a faster development and, consequently, lower yolk weight percentage at higher temperatures. We found positive relationships between incubation temperature and body size (length and weight) in the less developed stages, in which some yolk remained, but size decreased at increasing temperatures when yolk was completely exhausted.     

Surface disinfection of Atlantic halibut and turbot eggs with glutaraldehyde: evaluation of concentrations and contact times

To evaluate the effects of glutaraldehyde treatment at different disinfection temperatures, Atlantic halibut, Hippoglossus hippoglossus (L.), and turbot, Scophthalmus maximus (L.), eggs, incubated at 5 and 12°C, respectively, were disinfected with 400–1200 mg glutaraldehyde l-1 at three different contact times (2.5, 5 and 10 min). Egg batches of both poor and good quality were tested for halibut. Positive effects were more pronounced in poor than in good-quality batches at hatching. Egg disinfection had a highly positive effect on the viability of yolk-sac larvae in both types of batches. A level between 400 and 800 mg l-1 at a contact time of 5–10 min was optimal for halibut: at lower levels, the bactericidal effect was reduced, and at higher levels, there were indications of toxic effects. Halibut eggs disinfected with optimal doses of glutaraldehyde had furthermore a reduced hatching time and more synchronous hatching as compared with untreated eggs. Turbot was more sensitive to higher doses than halibut, and the best larval performance was obtained for 400–800 mg glutaraldehyde l-1 at a contact time of 2.5 min. A further evaluation should, however, be performed before recommendations are given for species incubated at temperatures higher than 5°C.