Purification,characterization and immunoassay of striped bass (Morone saxatilis) vitellogenin

The egg yolk precursor, vitellogenin (VTG), was purified from blood plasma of striped bass by chromatography on hydroxylapatite or DEAE-agarose. The fish were first implanted with estradiol-17β (E₂), which induced vitellogenesis. A rabbit antiserum (a-FSPP) raised against plasma from mature female striped bass, and then adsorbed with mature male plasma, was used to detect female-specific plasma protein (FSPP) in the chromatography fractions. Striped bass VTG (s-VTG) was collected from the peak fraction that was induced by E₂, reacted with a-FSPP, and contained all detectable phosphoprotein. It appeared as a single band (M_r ≂ 170,000) in SDS-PAGE or Western blots using a-FSPP, and as a pair of closely-spaced phospholipoprotein bands in native gradient-PAGE, suggesting that there is more than one circulating form of s-VTG. The relationship of s-VTG to the yolk proteins was verified using a-FSPP. The antiserum reacted with the main peak from gel filtration of saline ovary extracts, and it specifically immunostained the two main bands in Western blots of the extracts and the yolk granules of mature oocytes. The amino acid composition of s-VTG was similar to that of VTG from other fish and Xenopus. A radial immunodiffusion assay for s-VTG was developed using a-FSPP and purified s-VTG as standard. The s-VTG was not detected in blood plasma of males, immature females, or regressed adult females, but plasma s-VTG levels were highly correlated with plasma E₂ and testosterone levels, and oocyte growth, in maturing females. The results indicate that the maturational status of female striped bass can be identified by s-VTG immunoassay.     

Photoperiodic manipulations of the reproductive cycle of sea bass (Dicentrarchus labrax) and their effects on gonadal development, and plasma 17ß-estradiol and vitellogenin levels

The annual profile of plasma vitellogenin (VTG) and 17ß-estradiol (E₂) levels, as well as gonadal development and spawning characteristics were investigated in captive female sea bass (Dicentrarchus labrax). Endocrine and gonadal changes were studied in fish reared under natural conditions or exposed to manipulated photothermal cycles. In natural conditions of photoperiod and temperature, sea bass spawned from February through March (East coast of Spain, 40°N 0°E). One or two months of constant long-days (15L/9D) in a constant short-day photoperiod regime (9L/15D) all-year-round, given early in the year (March and March–April), advanced spawning by 3 months. The same treatment applied later in the year (September–October) delayed spawning by 1 month, compared to controls.In all groups, changes in plasma VTG levels were correlated with E₂ levels, oocyte growth and spawning time. In control females, VTG was low (<100 ng ml^-1) during the summer, until its first surge in plasma 4 months before the beginning of spawning. The VTG (3.1 ± 0.3 mg ml^-1) and E₂ (4.1 ± 0.5 ng ml^-1) levels showed a single annual peak during late vitellogenesis, the time of the highest proportion of vitellogenic oocytes in the ovary. Constant high levels of VTG (1–1.4 mg ml^-1) and E₂ (1.6–1.9ng ml^-1) were maintained during the entire spawning time, together with the presence of vitellogenic oocytes, suggesting the existence of several waves of oocyte growth in the ovary and thus, several spawns per female. Endocrine profiles and oocyte development in fish exposed to constant photoperiods were similar to controls, but were shifted in time in relation to the displacement of the spawning time. In the fish showing advanced spawns, the duration of the gametogenic proces was compressed when compared to controls. The differences observed in the evolution of the reproductive-related factors in the advanced groups, which were exposed to a reduction in temperature to 15°C, suggest an influence of the temperature in the early stages of the reproductive cycle in sea bass.     

Marine wax ester digestion in salmonid fish: a review

Alternative marine resources from lower trophic levels could partly cover the rapidly increasing needs for marine proteins and oils in the future. The North Atlantic calanoid copepod, Calanus finmarchicus, has a high level of lipids rich in n‐3 fatty acids. However, these animals have wax esters as the main lipid storage component rather than triacylglycerol (TAG). Although these esters are considered difficult to digest by many fish, is it well known that juvenile Atlantic salmon (Salmo salar) feed on zooplankton species. It is therefore possible that the capacity to utilize these lipids should be well developed in salmonids. Nonetheless, salmon hydrolyse wax esters slower than TAG and absorb fatty alcohols slower than fatty acids. However, salmon have several adaptations to digest diets rich in wax esters. These includes increased feed conversion, higher production of bile and higher activity of lipolytic enzymes in the midgut. Atlantic salmon has been shown to feed and grow on diets with a medium amount of wax esters (30% of the lipid) with results comparable to fish maintained on fish oil diets. Ingestion of higher level of wax esters (50% of the lipid) cause, however, poorer lipid digestibility and growth, so that optimal utilization of wax esters in Atlantic salmon is closer to 30% than 50% of the dietary lipid.     

The effect of tissue docosahexaenoic and arachidonic acids levels on hypersaline tolerance and leucocyte composition in striped bass (Morone saxatilis) larvae

Larval striped bass (M. saxatilis), tissue docosahexaenoic (DHA) and arachidonic (AA) acids levels were modulated through dietary enrichments and the effect on growth and survivorship examined. Mean growth was significantly greater in larvae enriched with AA than in larvae enriched with DHA (F-value for AA treatment was 20.5 versus only 5.1 for the DHA treatment). Dietary treatment did not have a significant effect on larval survivorship (56.0±2.4%, p>0.05). When challenged with hypersaline (25 psu) immersion, DHA enriched larvae survived better than AA enriched larvae, but larvae with body tissue levels of 15.4 mg AA g^–1 and 7.2–15.4 mg DHA g^–1 dry weight provided maximal survivorship to the challenge. Elevated levels of body tissue AA was generally associated with elevated levels of whole body cortisol. On the other hand, increasing levels of DHA mainly affected the kinetics of cortisol increase to hypersaline exposure. Larvae injected intraperitoneally with formalin fixed Staphylococcus aureus responded by altering the proportion of lymphocytes, monocytes and neutrophils in perpherial blood. Lymphocytes, which accounted for the largest percentage of white blood cells (over 70%), decreased in all challenged larvae during the first 6 hours post injection then returned to pre-challenge levels after 44 hours. Conversely, the relative proportion of monocytes and neutrophils rose from 14% and 2% up to 28% and 6% of the total circulating leucocytes, respectively. The largest increase occurred in larvae fed a moderate level of both DHA and AA.     

Preliminary investigations on the effects of dietary lipid on the spawning performance and egg quality of black sea bass Centropristis striata L

Adult black sea bass Centropristis striata broodstock ( N =162) were fed three different dietary treatments: two commercially prepared diets with 45% protein and two different lipid levels (12% and 20%) (diets 1 and 2), and a diet of frozen Atlantic silversides Menidia menidia (SS, diet 3). Broodstock were held under controlled photothermal conditions and induced to spawn with an LHRHa pellet (72 μg kg⁻¹ bw). Dietary lipid had pronounced effects on spawning performance and egg quality. Diet 3 (SS) produced a significantly ( P <0.05) higher fertilization success (22.4%) than diets 1 (0.6%) and 2 (4.8%). The hatching success of fertilized eggs was similar in all diets (range=40–58.6%), but only two spawns from diet 1 (12% lipid) yielded viable yolk-sac larvae (YSL). Diet 3 (SS) also produced significantly more YSL per female (21.8 × 10³) than the diet 1 (0.3 × 10³). Eggs from diet 3 (SS) contained a significantly greater proportion of n-3 series fatty acids, with docosahexaenoic acid (DHA) as the largest fraction. Eggs from commercially prepared dietary treatments contained significantly more n-6 fatty acids. The poor spawning performance of fish fed diet 1 (12% lipid) may be related to higher levels of linoleic acid and lower levels of DHA in the diet.     

Annual cycle of plasma insulin and glucose of sea bass.Dicentrarchus labrax,L.

Annual plasma insulin and glucose cycles were studied inDicentrarchus labrax maintained in either seawater (37.8) or brackish water (3.5). In both media, the highest insulin levels were found during the prespawning period (August–November) coincident to increases in weight and a decrease in plasma glucose. During spawning (December–April) and postspawning (May–July) periods, the decrease in insulin occurred at the same time as a reduction in growth and an increase of plasma glucose. Temperature and salinity conditions impeded spawning in the brackish water group, in which a minor weight loss was regained more quickly than in the sea water group; insulin levels were also higher.     

Comparative fatty acid composition of eggs from wild and captive black sea bass, Centropristis striata L.

Lipid content, lipid class and fatty acid compositions were determined in eggs from wild and captive black sea bass, Centropristis striata L., from northern (New England) and southern (South Carolina) regions to determine the effects of diet on egg composition and fertilization success. The formulated diets fed to the northern captive (NC) fish were higher in total lipids (22%) compared with the cut fish and squid diet fed to the southern fish (SC; 3.3% lipid) and had a higher relative amount of linoleic acid (LA) and lower relative amounts of docosahexaenoic acid (DHA) and arachidonic acid (AA). These dietary differences were broadly reflected in the egg lipid composition. The LA levels were higher in all lipid classes in eggs of NC fish while AA levels were lower. The DHA was higher in the major polar lipids (PL) (phosphatidylcholine and phosphatidylethanolamine) of SC eggs compared with NC. Compared with wild fish, both captive groups produced eggs with PLs richer in LA and lower in DHA and AA. Over all fish groups, fertilization success was directly correlated with levels of DHA and AA in the PLs and was inversely correlated with LA levels.     

Astaxanthin krill oil enhances the growth performance and fatty acid composition of the Pacific whiteleg shrimp,Litopenaeus vannamei,reared under hypersaline conditions

Hypersalinity culture of marine shrimp can lead to poor growth and feed efficiency. This study evaluated the effect of dietary supplementation of three oil sources (krill, fish and soybean) on the growth of Litopenaeus vannamei reared under high salinity. Shrimp of 2.79 ± 0.60 g were reared for 64 days under isosmotic (ISO, 23 ± 1.2 g/L) and hyperosmotic (HOS, 44 ± 2.0 g/L) conditions. Diets varied in their fatty acid composition: Control, 35 g/kg of the diet (as fed basis) soybean oil; Fish, 27 g/kg fish oil and 10 g/kg soybean oil; Krill, 48 g/kg krill oil and 4 g/kg soybean oil; Krill‐, 15 g/kg krill oil and 21 g/kg soybean oil; Krill+, 55 g/kg krill oil and 4 g/kg soybean oil. At harvest, Krill diet promoted the fastest shrimp growth (1.01 ± 0.01 g/week) and body weight (11.97 ± 2.01 g), regardless of water salinity. There were no significant differences in shrimp survival (93.4 ± 5.07%) and yield (554 ± 68.5 g/m²) among different diets. Shrimp fed Fish, Krill and Krill+ had higher concentrations of PUFA compared to those fed Control and Krill‐ diets.     

Sensory characteristics and nutritional value of red drum Sciaenops ocellatus reared in freshwater and seawater conditions

This study compares sensory attributes, chemical composition, fatty acid profiles, and the taste of raw and cooked red drum Sciaenops ocellatus fillets, reared in seawater (SW) and in freshwater (FW) aquaculture systems. Significant nutritional differences were found in the raw fillets. The total lipid was higher (5.31%) in FW fish than the SW (2.60%). The ratio n‐3/n‐6 and EPA/DHA was higher in SW than FW fillets. The eicosenoic acid was only present in FW fillets. DHA was dominant in SW specimens, contrary to the arachidonic acid level, which was dominant in FW fillets. The assessors perceived a significant difference in the firmness and colour of raw fillets, but its odour intensity was not affected. Fillets from SW fish have higher acceptability (33.3%) than FW fillets (26.4%). Sensory differences in raw fillet were not correlated with consumer perceptions, as cooked fish was considered to have similar flavour, independent of its origin.     

Effect of dietary essential fatty acids on level of oestradiol‐17β and vitellogenin,reproductive performance and larval quality of striped catfish (Pangasianodon hypophthalmus) in out‐of‐spawning season

A study was conducted to determine the effect of essential fatty acid in striped catfish (Pangasianodon hypophthalmus) broodstock diets on a level of oestradiol‐17β and vitellogenin, reproductive performance and larval quality in out‐of‐spawning season. Five female and five male broodstock were stocked into a net cage (3 × 5 × 1.5 m³) and fed with pelleted diets containing 1.5% fish oil and different amounts of corn oil (CO) in diet formulation, that is 0%, 1%, 2% and 3% respectively. Gonadal development, oestradiol‐17ß and vitellogenin concentrations were examined every 2 weeks. Oestradiol‐17ß and vitellogenin concentrations showed an increase during the maturation process. The significantly highest percentage of female mature, fecundity, hatching rate, larval production and larval survival rate were obtained from fish fed with 2% corn oil in the diet. This result revealed that the administration of 2% corn oil in the diet as a source of n‐6 (linoleic acid) and 1.5% fish oil as a source of n‐3 (linolenic acid) can improve the reproductive performance of striped catfish broodstock in out‐of‐spawning season.     

Effects of two diets on lipid composition and reproductive performance of brill (Scophthalmus rhombus) eggs

The effects of two diets with different lipid composition (squid Loligo gahi and one commercial pellet for breeders) on reproductive performance and egg lipid composition of brill (Scophthalmus rhombus) were compared. A total of 36 adult brill weighing 1482 ± 432 g were randomly distributed in the experimental tanks. A flow‐through system composed of four tanks with 4.2 m² of bottom area (5.0 m³ × 1.2 m water depth) was used. The experiment lasted for approximately 5 months (168 days). From the four mature females fed squid, three ovulated. Five females fed pellets matured and ovulated. Total ovulations were of 10 and 44, by the three females fed squid and the five ovulating female fed pellets respectively. Egg viability was higher, although not significantly, for females fed pellets. This indicates that pellets could be a better food for breeding females. In general, the egg total lipid content and the lipid classes composition did not seem to be affected by diet. In contrast, results indicate that not only diets but also individual females, regardless of the diet consumed, are determinant for egg fatty acid composition, clearly modulating their composition.     

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.     

Fatty acid and proximate composition of bluefin tuna (Thunnus thynnus) muscle with regard to plasma lipids

The composition of tail muscle fatty acids from wild and cultured bluefin tuna reared on a diet based on herring and sardine, along with the plasma lipid profile of the farmed individuals, was determined. The total lipid content of farmed bluefin in this study was 0.922 g/100 g or 3.49 g of saturated fatty acids (SAFA), 4.48 g of monounsaturated fatty acids (MUFA), 2.58 g polyunsaturated fatty acids (PUFA) n‐3 and 0.37 g of PUFA n‐6 fatty acids; for wild specimens, it was 0.920 g/100 g, or 2.85 g of SAFA, 4.82 g of MUFA, 2.78 g PUFA n‐3 and 0.27 g of PUFA n‐6 fatty acids. The major fatty acids in this study were 16:0; 16:1, n‐7; 18:1, n‐9 and DHA 22:6, n‐3 acids. The sum of these major components accounted for more than 57% and 80% of the total fatty acids in all the samples of farmed and wild tuna respectively. No significant differences in the proximate composition were demonstrated between farmed and wild samples, except for the energy value, in favour of the farmed tuna. Statistically, glucose tends to increase together with cholesterol (CHOL) and plasma triglyceride, as for these pairs, it showed positive correlation coefficients and P>0.05. Some measured tuna metabolites demonstrated strong mutual correlations, especially GLU, CHOL and TRIG, which are crucial factors in the lipid profile of animals.     

瓯江彩鲤生殖周期内卵巢成熟系数及脂类、脂肪酸的变化

采用生化方法测定了瓯江彩鲤(Cyprinus carpio var.color)卵巢不同发育时期的成熟系数及卵巢脂类、脂肪酸含量与组成,旨在了解瓯江彩鲤卵巢中脂类、脂肪酸的变化与卵巢发育及卵细胞成熟之间的关系。结果显示:卵巢中总脂含量在产卵前最高,占干重17.62%,产卵后显著下降到12.84%(P<0.05),到快速生长期又出现稍微下降,之后在越冬期开始呈上升趋势。4个时期中,C16∶0(软脂酸)、C18∶1n-9c(油酸)、C18∶2n-6c(亚油酸)均是卵巢中的主导脂肪酸,其含量显著高于其它脂肪酸(P<0.05);产卵前,卵巢中脂肪酸含量高,但种类较少,产卵后,卵巢中脂肪酸含量低,但种类较多;随卵巢的发育,不饱和脂肪酸的比例增加,从快速生长期到产卵前,C20∶4n-6(ARA)与C22∶6n-3(DHA)的含量逐渐上升,最高分别达11.63 mg/g、17.80 mg/g。结果表明:不同时期卵巢中脂类含量与成熟系数呈正相关,卵巢成熟需积累较多脂类物质。不饱和脂肪酸尤其是DHA、ARA对卵巢的成熟具有重要作用。     

Reproductive performance, lipids and fatty acids of mud crab Scylla serrata (Forsskål) fed dietary lipid levels

Natural food (NF, control), artificial diets (AD) containing total lipid levels of 10%, 12% and 14% (AD10, AD12 and AD14) and their combinations (AD10+NF, AD12+NF and AD14+NF) were fed for 112 days to pond‐sourced eyestalk‐ablated mud crab Scylla serrata (625±6.4 g) in tanks in order to determine their effects on reproduction and lipid profiles in broodstock tissues and zoeae. Crabs fed NF had the highest number of spawning followed by crabs fed AD10+NF and AD14+NF. Higher offspring production (number of zoeae) was obtained from crabs fed NF and AD+NF than from AD. As dietary total lipid levels increased, total lipid of broodstock ovaries, hepatopancreas, muscle and zoeae correspondingly increased in which AD+NF promoted higher levels than AD. Increased dietary total lipid levels enhanced lipid classes such as triacylglycerols and phosphatidyl choline levels in zoeae, all higher in crabs fed AD+NF than in AD. The major fatty acids in zoeae, particularly 16:0, 18:0, 18:1n‐9 and 20:4n‐6, 20:5n‐3 and 22:6n‐3, were higher in crabs fed AD+NF than in AD, the contents corresponding to broodstock dietary total lipid levels. A 10% total lipid in AD in combination with NF was sufficient to provide the essential lipids in crabs in the improvement of larval production and quality.     

Could a diet enriched with n‐3 highly unsaturated fatty acids be considered a promising way to enhance the immune defences and the resistance of Penaeid prawns to environmental stress?

The prawn Penaeus stylirostris (Stimpson), when fed for 28 days with n‐3 highly unsaturated fatty acid (HUFA)‐enriched feed pellets, demonstrated an enhanced resistance to variations in environmental parameters (a decrease in temperature and salinity over a 4‐day period from 28 °C to 17 °C and from 35‰ to 10‰ respectively) and an improvement in their immune defence potential, i.e. increased agglutination titre of plasma and increased respiratory burst of haemocytes.     

Quality of eggs and spermatozoa of rainbow trout fed an n-3 essential fatty acid-deficient diet and its effects on the lipid and fatty acid components of eggs, semen and livers

ABSTRACT: The effect of an n-3 essential fatty acid (EFA) -deficient diet on spawning and on the lipid and fatty acid contents of eggs, semen and livers of rainbow trout Oncorhynchus mykiss was investigated. Fish were split into two groups and fed either of two diets for a period of 4 months prior to the start of the spawning season. The control group was fed a commercial diet, containing n-3 EFA, whereas the experimental group was fed an n-3 EFA-deficient diet. Fish were 3 years old at the time of spawning. Eggs and semen were stripped off five females and five males from each diet group and cross-fertilized. Two of the five males fed the deficient diet showed a lower sperm motility, resulting in slightly lower mean hatching rates when crossed with eggs of either group. Higher lipid contents in the EFA-deficient diet were reflected in the egg and semen lipid contents, whereas the lipid contents of male livers of both diet groups were higher than those of female livers. In livers and eggs, the main polar lipid was phosphatidylcholine accompanied by phosphatidylethanolamine in sperm polar lipids. The non-polar lipids of eggs were mainly triacylglycerols but in livers and semen, free fatty acids and free sterols were more abundant. Essential fatty acids, namely n-3 highly unsaturated fatty acids and linolenic acid, were generally lower in eggs, semen and livers sampled from the EFA-deficient diet group. These results indicate that the fertilization, eyed and hatching rates obtained from crossings with males fed the EFA-deficient diet were slightly lower because of the dietary effect on sperm motility. Moreover, lipids of eggs, semen and livers of male and female rainbow trout were influenced greatly by their dietary availability.     

Abundance and origins of plasma lipids in sexually maturing female coho salmon Oncorhynchus kisutch (Walbaum) in culture

Coho salmon Oncorhynchus kisutch are semelparous spawners with highly synchronized oocyte growth. Plasma was collected from maturing female fish and plasma lipids were quantified by lipoprotein class. Stable ¹³C isotopes were employed to investigate the origins of plasma lipids. Plasma lipoproteins were partitioned into the very low density (VLDL), low density (LDL), high density (HDL) and very high density (VHDL) lipoprotein classes. Lipids from all lipoprotein classes increased between the lipid droplet and early vitellogenesis stages of oocyte growth and VHDL lipids continued to increase through mid vitellogenesis (P < 0.05). During vitellogenesis, total plasma lipid concentrations were similar to the sum of lipoprotein lipids (P = 0.51). Plasma vitellogenin determined by an ELISA method was very well correlated with VHDL lipid during vitellogenesis (R² = 0.91, n = 15). In general, the δ¹³C values of plasma lipids reflected that of feed (exogenous) and muscle (endogenous) lipids when feed and muscle δ¹³C values were similar, and were intermediate when differences existed between the two lipid sources. With one exception, the δ¹³C values of lipids from all lipoprotein classes within a plasma sample were similar. Results indicate that during sexual maturity, lipids from all plasma lipoprotein classes originate from a common pool of exogenous and endogenous lipids.