Fatty acids and egg quality in great scallop

Spawners of the great scallop, Pecten moximus, were conditioned on three microalgal diets: T-Isochrysis, a mixture of four species (PTSC) and Chaetoceros calcitrans.The polyunsaturated fatty acid (PUFA) composition of neutral and polar lipids of eggs was related to the fatty acid composition of the diet. However, the 20 and 22 carbon PUFAs were maintained at significant levels independent of those of the diet when these fatty acids were present in the diet at low levels. This effect was more pronounced in the polar than in the neutral lipids. A preferential incorporation of 22:6(-3) and 20:4(-6) was demonstrated in the polar lipids. This emphasizes their role in ovogenesis and embryogenesis.The 22:6(-3) requirement of P. maximus was better satisfied by T-Isochrysis, which favoured the highest incorporation of this essential fatty acid in the eggs. The good success in reproduction obtained with this monospecies diet led us to modify multispecies diets by increasing the level of 22:6(-3).     

Effects of steroid hormones on in vitro oocyte maturation in white sturgeon (Acipenser transmontanus)

The in vitro effects of several steroids on the maturation of intact white sturgeon (Acipenser transmontanus) ovarian follicles were investigated. At the highest concentration (1024 ng ml^–1 for the C21 steroids and 1139 ng ml^–1 for the C19 steroids), all of the C21 steroids tested, progesterone (P4), 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,(20,21-trihydroxy-4-pregnen-3-one 20-S), 11-deoxycortisol (S) and cortisol (F), as well as testosterone (T) induced germinal vesicle breakdown (GVBD) at 14 and 22 h. At 6 h, only P4 and 17,20-P induced maturation at the highest concentration (1024 ng ml^–1). At 14 and 22 h, 11-deoxycortisol was the most potent steroid inducer of GVBD followed by P4, 17OHP, 17,20-P, and 20-S. The steroid 11-hydroxytestosterone (11OHT) was completely ineffective at all concentrations and exposure times. The C21 steroids induced oocyte maturation at concentrations ranging from 4 to 1024 ng ml^–1, whereas T induced GVBD at 225 to 1139 ng ml^–1. Calculation of the mean effective concentration that induced 50% GVBD (EC50) from the 22 h incubations revealed the following order of potencies: S > P4 > 17OHP > 17,20-P > 20-S >> F > T. These bioassay results, together with previous findings on the endogenous production of steroids by ovarian follicles from gonadotropin-primed females, indicate that more than one steroid has a biological role in the resumption of meiosis in sturgeon oocytes and provides empirical evidence for P4, 17OHP, S, 20-S, and 17,20-P as maturation-inducing steroids in white sturgeon.     

The nature of hematological response in fish

Hematological status was examined in rainbow trout,Oncorhynchus mykiss, held for 3–4 weeks under temperature, photoperiod and P_{O 2} conditions approximating those of their winter, spring and summer habitats. The most striking change observed was in red cell population composition. In winter fish mature cells were predominant; juvenile and developing erythrocytes characterized spring and summer animals. Hemoglobin, hematocrit and both mean erythrocytic volume and hemoglobin were modestly lower in spring and summer than in winter fish. Red cell numbers were not significantly affected. These observations suggest that avoidance of viscosity-based increases in circulatory work cost is more advantageous than elevation of blood O₂-carrying capacity. Although hemoglobin isomorph profiles were significantly altered, there is little evidence that such changes are of critical adaptive importance. Given presumed age-based reduction in gas transport effectiveness, the replacement of mature and senescent cells by more metabolically-competent juvenile cells appears to be the pivotal event in hematological response. Leucocyte counts were significantly elevated in spring and summer as compared to winter fish. Lymphocyte/heterophil ratios declined from 8.27 in winter fish to 3.13 in summer trout. Thrombocyte, monocyte, eosinophil and basophil abundances were little changed.     

Influence of lipid composition of live feed on growth,survival and pigmentation of turbot larvae

The effect of different lipid compositions of live feed on the survival, growth rate and pigmentation success of turbot larvae, Scophthalmus maximus (L.), was investigated. Rotifers, Brachionus plicatilis, together with the algae Tetraselmis sp., were administered until day 12, and Artemia was fed until day 27. The experimentally treated live feeds were enriched with four formulated emulsions, resulting in a gradient in the relative contents of 3 HUFA (highly unsaturated fatty acids) and in DHA (docosahexaenoic acid, 22:6 3)/EPA (eicosapentaenoic acid, 20:5 3) ratios in both the rotifers and Artemia.There were no differences in larval growth rate, and only small differences in survival rate throughout the feeding experiment, probably because of satisfactory levels of 3 HUFA in the live feed to sustain growth and survival. A correlation was obtained between the percentage of completely pigmented 27 d old turbot and the DHA/EPA ratio in the total lipids of 12 d old larvae, which again was correlated with the corresponding ratio in the live feed used. The results suggest that normal pigmentation in turbot requires dietary DHA in the early larval feeding period, and that this requirement cannot be replaced by EPA.     

Effect of feeding schedule on growth of rainbow trout

Rainbow trout (Oncorhynchus mykiss) fingerlings were reared for 2 months under one of two different feeding regimes (one and two diurnal peaks) and under a constant feeding regime as a control in order to find out if it is possible to fit the feeding to the assumed circadian rhythm. The differences in the specific growth rates observed between the different feeding treatments were not significant, but the tank-effect and the effect of the genetic background of the fish were highly significant. The conclusion that no beneficial effects are achievable through alteration of the feeding regime and the significance of a possible genotype-feeding-regime interaction are discussed.     

A preliminary study of the behaviour and vitality of reseeded juvenile great scallops, of three sizes in three seasons

In order to have a better understanding of recessing in great scallop, Pecten maximus and consequently the causes of mortality at reseeding, this study has monitored, at different seasons, the dispersion and recessing of different sizes of juveniles (about 15, 30 and 45 mm, called small, medium and large) after seeding. Moreover, the aim was to see when small spat (15 mm) could be seeded, and thus reduce the costs of intermediate culture.Three monitoring approaches were used together: (1) continual observations by remote video camera, of a defined area (less than 1 m²) containing 10 scallops from each size group; (2) daily monitoring of behaviour with divers along three bottom lines, with 20 × 1 m² plots each and nine marked scallops per plot; and (3) the biochemical content of the muscle: adenylic energetic charge and storage of energy reserves (glucides, proteins, lipids).The video monitoring identified but did not quantify predator behaviour, particularly at night. The role and behaviour of spiny crab, Maia squinado, and of small predators has clearly been shown, such as: (a) small crustaceans, Inachus sp., breaking the edges of scallop valves; and (b) small gobies, Pomatoschistus pictus, pecking the tentacles of the scallop mantle.For the monitoring by divers, filtering appeared much too difficult to look at for it was very disturbed by divers, and anyway the resumption of filtering came immediately after seeding. On the other hand, diver monitoring of dispersal and recessing was quite easy to do with a minimum of practice. On the basis of dispersal, the best seasons for seeding appear to be spring or summer. In autumn, two-thirds of small and medium juveniles are missing 3 days after seeding, but we could not observe whether they had been eaten by predators or had just moved and recessed farther. There was no experiment in winter owing to adverse conditions for scallop seedings.Biochemical analyses confirmed the unsuitability of autumn for scallop seeding, because of very low glucide content in this season.The adenylic energetic charge in the smooth part of the muscle showed that stress before seeding (aerial exposure, handling), and post-seeding behaviour (swimming, recessing) have a high energetic cost for scallops. In summer and autumn, 3 days after seeding, none of the three size batches recovered their initial vitality.     

Cultivation of the erizo: an evaluation of eggs and postmetamorphic juveniles size selection

Evidence in the course of experiments under controlled conditions proved that the egg and juvenile characteristics of the edible Chilean sea urchin, erizo (Loxechinus albus) influence the cultivation efficiency. Mortalities among larvae originating from eggs with a diameter <110 m were higher than among those originating from eggs with a diameter >100 m. On the other hand, the initial size of the egg did not produce any variation in the percentage of larvae which metamorphosed. The initial size of the late metamorphosing juveniles modified early growth. A direct relationship was established between the size of the juveniles and the growth rate. In conclusion, biological aspects are determinant factors in any consideration of options available for the mass production of larvae and juveniles of this sea urchin.     

Quantitation of inert feed ingestion in larval silver sea bream (Sparus sarba) using auto-fluorescence of alginate-based microparticulate diets

The ingestion of an inert feed as a sole food source was investigated in larval silver sea bream (Sparus sarba) fed an alginate-based microparticulate diet. Using the auto-fluorescent properties of pigments associated with the alginate base, ingestion and gut content were investigated over a 6 h experimental period in fed and unfed larvae. By extracting and measuring chlorophyll a (Chl a) and phaeopigment content of feeding larval fish and relating this to standardized Chl a and phaeopigment content of the diet, relative to diet weight, it was determined that individual fed 7-day old larvae had a maximum gut content of 1.05±0.09 g diet while 14-day old fed fish had a maximum gut content of 3.17±0.90 g diet. On average, the gut content of 14-day old fish was 2.89 times greater than the gut content of 7-day old fish. The dry weight of larval sea bream increased from 43±4.2 g at day 7 to 134.3±20.4 g at day 14 indicating that growth of fish fed this inert feed was substantial. Gut pigment dynamics suggested that Chl a was degraded to phaeopigments by 7-day but not 14-day old larvae and the individual gut dietary content varied considerably in 14-day old fish. The maximum Chl a and phaeopigment content in larval sea bream was 0.4 ng ind^–1 and 0.55 ng ind^–1 for 7-day old fish and 1.54 ng ind^–1 and 2.81 ng ind^–1 for 14-day old fish respectively. The present method may potentially allow simple and direct assessment of larval fish feed ingestion in both an experimental and commercial setting.     

Changes in amino acids and essential fatty acids during early larval rearing of dentex

Samples of fertilized dentex, Dentex dentex, eggs and larvae fed enriched rotifers and Artemia according to standard hatchery procedures were analysed for free, total amino acid and fatty acid contents. Egg free amino acids (53 nmol ind-1) and total lipids (13.5 (g ind-1) levels were considerably reduced in the newly hatched larvae (6.0 nmol and 5.7 (g ind-1 respectively) while the amount of -3 PUFA (Polyunsaturated fatty acid) and 20:4-6 remained unchanged suggesting their possible role as essential fatty acids (EFA) in this species. In general, the changes in pattern of the major biochemical components during early development of dentex was similar to that outlined for other Sparidae species investigated to date. The essential amino acid profile of enriched live food did not show major imbalances when compared to that of dentex larvae. In contrast, the level of docosahexaenoic acid (22:63, DHA) and the DHA:EPA (eicosapentaenoic acid, 20:5-3) ratio in larvae given rotifers and Artemia were significantly lower relative to the corresponding values in the unfed larvae.     

Oocyte maturation inClarias batrachus. III. Purification and characterization of maturation-inducing steroid

Maturation-inducing steroid (MIS) in the Indian female catfish,Clarias batrachus, was purified and characterized from the incubation medium in which fully grown but immature folliculated oocytes were incubated with salmon gonadotropin (SG-G100) for 36 h. Maturation-inducing (MI) activity of residues obtained at various steps of extraction and purification was assessed byin vitro germinal vesicle breakdown (GVBD) assay using folliculated oocytes ofC. batrachus. The post incubation medium was extracted with diethyl ether. The ether phase was partitioned using 50% methanol plus n-hexane. The methanol phase which had MI activity was fractionated into 7 fractions using reverse-phase high-performance liquid chromatography. Of these 7 fractions, fraction 3 was found to be active in having MI ability and identified as 17 ,20-dihydroxy-4-pregnen-3-one (17,20-diOHprog). The authenticity of 17,20-diOHprog as the major follicular mediator of gonadotropin-induced oocyte maturation was further confirmed by thin-layer chromatography (TLC) in which fraction 3 was run along with authentic 17,20-diOHprog standard. This investigation gives a direct evidence that 17,20-diOHprog is the major naturally occurring MIS in Indian female catfish,C. batrachus.     

Steroid release from separated theca and granulosa layers of Atlantic salmon (Salmo salar) ovarian follicles: the effects of a purified salmon gonadotrophin preparation

Theca and granulosa layers were removed from ovarian follicles of mature Atlantic salmon (Salmo salar) and were separately incubated under sterile conditions with and without a partially purified salmon gonadotrophin preparation (GTH). Aliquots of the incubation media were removed at intervals and analysed for the steroids 17, 20-dihydroxy-4-pregnen-3-one (1720P), 17-hydroxyprogesterone, progesterone, androstenedione, testosterone and oestradiol. The biosynthesis of C19 and C21 steroids was very largely restricted to the thecal tissue and was markedly stimulated in the presence of GTH. Androstenedione (max 65 ng/ml) and testosterone (max 14 ng/ml) were released from the earliest stages of incubation whereas the release of 17-hydroxyprogesterone (max 51 ng/ml) and progesterone (max 5.5 ng/ml) commenced only after a lengthy induction period. A trace (1.0 ng/ml) of 1720P was produced by the theca in the presence of GTH but oestradiol was not detected. The granulosa preparations released levels of 17-hydroxyprogesterone and androstenedione only marginally above the detection limits (ca 0.7 ng/ml) and there was little stimulation of output with GTH. Oestradiol (max 4 ng/ml) was released only in the presence of GTH. 1720P, progesterone and testosterone were not detected as products of this tissue. These results, together with those derived earlier from incubations of complete follicles support the view that the synthesis of 1720P is essentially a two-cell process in which 17-hydroxyprogesterone produced in the theca is subject to the action of steroid 20-hydroxysteroid dehydrogenase in the granulosa. The temporal pattern of release of steroids in these and earlier experiments is considered in relation to mechanisms of steroid biosynthesis and to their possible roles in oocyte final maturation.     

Evaluation of vitamin C-enriched Artemia nauplii for larvae of the giant freshwater prawn

The effect of high levels of ascorbic acid (AA) delivered through enriched live food has been verified through the successful culture of larval giant freshwater prawn, Macrobrachium rosenbergii. Two successive feeding trials were set up using a control (550 g AA g^–1 DW) and two different AA-enrichment levels in Artemia (1300 and 2750 g AA g^–1 DW). Under standard culture conditions, no differences in growth nor survival could be observed demonstrating that the nutritional requirements are below 550 g AA g^–1 DW, which is the normal level occurring in freshly-hatched Artemia. However, a significantly positive effect could be demonstrated on the physiological condition of the postlarvae, measured by means of a salinity stress test, when vitamin C-boosted live food was administered. Since the AA levels in the predator larvae are linked with the enrichment levels in the live prey, it may be assumed that a positive influence on stress resistance was caused by feeding vitamin C-enriched Artemia. It is expected that under suboptimal conditions, supplementation of high vitamin C levels might also enhance production characteristics.     

Dehydration is more effective for the control of embryonic development and larval hatching of Diplectanum aequans (Monogenea, Diplectanidae) than formalin and trichlorphon

Embryonic development and larval hatching of the monogenean Diplectanum aequans, gill parasite of sea bass Dicentrarchus labrax, was studied in relation to different prophylactic treatments. Groups of eggs of D. aequans were submitted to different in vitro treatments: formalin (300 and 100 L L^–1 per 1 hour), Neguvon^® (trichlorphon 0.2 mg L^–1 per 48 hours) and dehydration for 4 and 8 hours. Percentages of hatched larvae, aborted larvae and undeveloped embryos were estimated in comparison with the control group. Results showed that 300 L L^–1 formalin and dehydration treatments were able to reduce larval hatching significantly, while Neguvon^® and 100 L L^–1 formalin treatments had no effect.     

Incorporation and metabolism of 14C-labelled polyunsaturated fatty acids in juvenile gilthead sea bream Sparus aurata L. in vivo

The incorporation, and the capacity for desaturation and elongation in vivo, of intraperitoneally-injected, ¹⁴C-labelled n–3 and n–6 C₁₈ and C₂₀ PUFAs were investigated in juvenile gilthead sea bream, Sparus aurata. The results indicate that juvenile gilthead sea bream have only limited ability to convert CH PUFAs to C₂₀ and C₂₂ HUFAs in vivo. The data are consistent with the results from nutritional studies on larvae, postlarvae and fingerlings that have shown that gilthead sea bream require the provision of preformed eicosapentaenoic and docosahexaenoic acids in the diet. The impairment in the desaturase/elongase pathway was quantitatively and qualitatively similar to that found in turbot, Scophthalmus maximus, being at the level of the 5-desaturase. The low activity of 5-desaturase combined with the consistent finding that arachidonic acid is selectively retained in membrane phosphatidylinositol suggests that, in addition to eicosapentaenoic and docosahexaenoic acids, gilthead sea bream may also have a requirement for preformed arachidonic acid in the diet.Abbreviations AA 5,8,11,14-eicosapenaenoic acid (arachidonic acid, 20:4n–6) - CPL diradyl (diacyl + alkenylacyl + alkylacyl) glycerophosphocholine - DHA 4,7,10,13,16,19-docosahexaenoic acid (22:6n–3) - EPA 5,8,11,14,17-eicosapentaenoic acid (20:5n–3) - EPL diradyl (diacyl, alkenylacyl + alkylacyl) glycerophosphoethanolamine - HUFA highly unsaturated fatty acids ( C₂₀ and with 3 double bonds) - LA 9,12-octadecadienoic acid (linoleic acid, 18:2n–6) - LNA 9,12,15-octadecatrienoic acid (-linolenic acid, 18:3n–3) - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid(s)     

Effects of feeding (ω-3) HUFA-enriched Artemia during a progressively increasing period on the larviculture of freshwater prawns

Larvae of the freshwater prawn (Macrobrachium rosenbergii) were reared to post-larvae in a closed recirculation system. Six treatments were designed where freshly-hatched Artemia nauplii were first given to the larvae for 3, 7, 11, 15, 17 and 25 days according to the treatment. After that (-3) HUFA-enriched Artemia were given until the 28th day at which time the test was terminated and evaluated. The requirement for (-3) Highly Unsaturated Fatty Acids (HUFA) for the larval stages of M. rosenbergii was confirmed by this experiment. Moreover, the longer the period of feeding on (-3) HUFA-enriched Artemia nauplii, the better the results in terms of growth, metamorphosis rate, survival and stress resistance.     

Genetic improvement of farmed tilapias: Biochemical characterization of strain differences in Nile tilapia

Four African wild strains (Egypt, Ghana, Senegal and Kenya) and four established Asian farmed strains of Nile tilapia, Oreochromis niloticus (popularly known in the Philippines as Taiwan, Thailand, Singapore and Israel) were analysed electrophoretically at 30 protein loci to estimate genetic differences among the strains. All strains shared alleles at 14 monomorphic and 16 variable loci. Among the African strains, characteristic allele frequency differences were observed at AAT-1 ^* 46 for Ghana and Senegal, ADH ^* 83 for Kenya, ADH ^* 120 for Senegal, G3PDH-2 ^* 300 for Egypt, IDDH ^* 67 for Senegal, sMDH-1 ^* 120 for Kenya and SOD ^* 150 for Senegal. Genetic distance values among the strains revealed a clustering of the farmed strains with Egypt and Ghana O. niloticus, a slight separation of the Senegal strain and a larger separation of the Kenya strain. This profile may reflect the origins of the few founder populations of this species previously introduced to Asia. It also confirms the wider genetic divergence of the Kenya strain (O. niloticus vulcani) from the others studied here, which are all O. n. niloticus. Observed heterozygosities of the strains ranged from 0.026 to 0.071, with the African wild strains the lower values (mean Ho = 0.036) and the farmed strains the higher ones (mean Ho = 0.056). The implications of these results to the ongoing tilapia genetic improvement programme in the Philippines are discussed.     

Bacteria in the culture of the scallop Argopecten purpuratus (Lamarck, 1819)

The scallop Argopecten purpuratus is one of themajor resources of commercial importance in Chilean aquaculture. One of themajor limiting factors for the culture of this species has been mass mortalityoccurring during production of juvenile seed organisms where massmortality of veliger larvae has been attributed to the presence of pathogenicbacteria in scallops and hatchery systems. Bacteriological studies havedemonstrated that in addition to bacterial pathogens, beneficial bacterialspecies capable of improving scallop larval survival may also be isolated fromscallops and hatchery systems. Research has been carried out on thedetermination of the feasibility of implementing beneficial bacteria in thecontrol of culture pathogens, thus reducing the need for chemotherapeuticmethods. The present review analyzes bacteriological research data on thedifferent roles of bacteria associated with scallop culture and discussesrecentdata on the implementation of beneficial bacteria in biological control ofpathogens in larval mass cultures of Argopectenpurpuratus.     

Progestogen metabolism by ovaries of the roach (Rutilus rutilus L.) and the rudd (Scardinius erythrophthalmus L.)

Roach ovaries converted 17-hydroxyprogesterone to 17,20-dihydroxy-4-pregnen-3-one (17,20P) and to glucuronides of testosterone and 17,20P. Small amounts of 5-pregnane-3- and -3, 17, 20-triols, 7-hydroxy-5-reduced metabolites and 17,20-dihydroxy-4-pregnen-3-one (17,20P) were also formed. Rudd ovaries converted this substrate mainly to 17,20P, 5-pregnane-3- and -3,17,20-triols, 17,20-dihydroxy-5-pregnan-3-one and testosterone glucuronide. The main metabolites of progesterone with both species were 17,20P, 5-pregnane-3,17,20-triol and 7-hydroxy-5-reduced steroids. Rudd ovaries formed, in addition, 17,20-dihydroxy-5-pregnan-3-one from progesterone. The pattern of metabolites was markedly altered when the concentration of substrate was increased from 42ng to 1 µg or 100 µg. At the highest concentration, glucuronides and polar steroids were not detectable, while at low concentrations they accounted for over 50% of the metabolites. 20-Hydroxysteroid dehydrogenase was shown to have a very high capacity, producing 21–47 µg 17,20P from 100 µg 17-hydroxyprogesterone substrate with 200 mg ovarian tissue in 5h.     

The effect of seawater flow and temperature on metamorphosis and postlarval development in great scallop

Variations in growth and survival of hatchery-reared post-metamorphicjuveniles of great scallop Pecten maximus prompted anexamination of settlement and postlarval development. The effects ofseawater flow and temperature on great scallop metamorphosis andpostlarvae were studied over a 4–5 week period. In allexperiments, and regardless of environmental conditions, great scallopmetamorphosed after a 2–3 week period with values of 35 to70%. Subsequently, spat numbers increased slightly. Spatmortality generally occurred from the third week onward and reachedlevels as high as 30% by the fifth week under standardconditions. At 20 °C, however, 60% mortality levels wererecorded. Differences in spat growth rate, ranging from 37 to 45 mday^–1, were noticed at different seawater flow ratesbut no clear tendency could be discerned. Temperature affected spatgrowth with an increase in size from 24 m day^–1 at15 °C to 35 m day^–1 at 18 °C. Conversely,growth was suppressed at 20 °C (14 m day^–1).For optimal metamorphosis and postlarval development in great scallop, aseawater flow of 4.3 L h^–1 per sieve and a temperatureof 15 °C are recommended.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.