Vitamin requirements in great scallop larvae

Vitamins were analysed in food (microalgae) and larvae of great scallop, Pecten maximus, during larval development. Microalgae used to feed larvae in hatcheries show great variability in their vitamin composition depending on both the species and culture condition (phase of growth). The microalgae used to feed Pecten maximus larvae were rich in vitamins; their content compared with diets used in fish culture appeared sufficient, with the possible exceptions of pyridoxine, biotin and pantothenic acid. Vitamins in bacteria, isolated from the larval rearing tank were also analysed, as they can also contribute to the diet. Vitamin B₁₂, -tocopherol and -carotene were detected in very low concentration in bacteria; however, some bacterial strains were rich in pantothenic acid, and the pattern of other vitamins was similar to that from microalgae. The presence of bacteria can complement the diet in panthothenic acid, as it has been demonstrated that bacteria are ingested by larvae. The vitamin content of Pecten maximus larvae was analysed from the second day after hatching to just before metamorphosis. The content of some vitamins, ascorbic acid (C), -tocopherol and -carotene, increased during larval development, suggesting that their requirement was satisfied. However, thiamin and riboflavin decreased during larval development and further studies, possibly using microencapsulated vitamins supplements, are needed to determine whether these vitamins are limiting during larval development.     

Seasonal changes in reproductive condition and plasma levels of sex steroids in the blue cod,Parapercis colias (Bloch and Schneider) (Mugiloididae)

Gonad and plasma samples were taken from blue cod captured throughout the reproductive cycle, gonad condition was assessed, and plasma levels of 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20P), testosterone (T), 17-estradiol (E₂) and estrone (E₁) were measured by radioimmunoassay. It was confirmed that spawning occurred over an extended period in late winter and spring, with individual fish being involved in multiple spawning events. Plasma levels of T were bimodal in both sexes with peaks (maximum of 6.0 ng.ml^–1) occurring 2 months prior to, and also during the early part of the spawning period. 17,20P was elevated in males (2.1 ng.ml^–1) in mid-spermatogenesis coinciding with the first T peak (4.9 ng.m.^–1). 17,20P was detectable but not significantly elevated (0.6–1.2 ng.ml^–1) at any sample time in females. E₂ was elevated in mature females (1.0 ng.ml^–1) early in the spawning period but remained at assay detection limits (0.3 ng.ml^–1) at all other sample times. Neither 17OHP nor E₁ were detectable in the plasma of either sex. It is suggested that bimodal increases in sex steroids prior to spawning may be a feature of species with rapid recrudescence.     

Regulation of oocyte maturation in the rainbow trout,Salmo gairdneri: role of cyclic AMP in the mechanism of action of the maturation inducing steroid (MIS), 17α-hydroxy, 20β-dihydroprogesterone

In fish, oocyte maturation (resumption of meiosis after completion of vitellogenesis and before ovulation) is triggered by maturation inducing steroids (MIS) which generally appear to be secreted in the ovary in response to stimulation by a pituitary maturational gonadotropin. Converging data from different laboratories show that 17-hydroxy, 20-dihydroprogesterone (17, 20-OH-P) is the principal MIS in salmonoids; but clear identification remains to be done in other taxonomic groups.The experiments reported here in the rainbow troutSalmo gairdneri examine the possible involvement of oocyte cAMP on the mechanism of MIS action. The action of 17, 20-OH-P, on germinal vesicle breakdown (GVBD) in oocytes incubatedin vitro within the follicle, was inhibited by various substances expected to elevate the intraoocyte concentrations of cAMP: cAMP ( 1 mM) or dibutyril cAMP ( 2 mM), phosphodiesterase inhibitors such as theophylline ( 0.2 mM) or 3-isobutyl-1 methylxanthine (IBMX 0.1 mM), adenylate cyclase activators such as cholera toxin (> 100 nM) or forskolin ( 0.03 mM). In fact, the combined action of IBMX (1 mM) and forskolin (0.01 or 0.05 mM)in vitro was to promote accumulation of intraoocyte cAMP within 1 to 5 hours. Oocyte cAMP concentrations exhibited a large variability between different females, depending on the stage of oocyte development; a significant positive correlation between oocyte cAMP concentration and the follicular weight, and a significant negative correlation between oocyte cAMP concentration and the median efficient dose of 17, 20-OH-P for induction of GVBD, were observed. Finally, when intrafollicular oocytes were incubatedin vitro, the addition of a maturation-inducing concentration of 17, 20-OH-P (3×10^–6M) induced a significant decrease of oocyte cAMP within the first 10 hours of incubation. These results show that cAMP appears to play a central role in the regulation of oocyte sensitivity to 17, 20-OH-P and in the intraoocyte mechanisms leading to GVBD in trout.These data are discussed together with the few indications available in fish concerning the mechanism of MIS action which can be compared to some extent with the amphibian model.     

Variation in content of Ω3 fatty acids in farmed Atlantic salmon,with special emphasis on effects of non-dietary factors

The main aim of the present study was to examine the impact of some biological and environmental factors on the lipid and fatty acid compositions of farmed Atlantic salmon (Salmo salar), with special emphasis on 3 fatty acids. Two year groups of salmon at nine fish farms distributed along the Norwegian coast were fed the same diet and were sampled every second month. The data are believed to give a representative characterization of lipid and fatty acid content of salmon farmed in Norway.Multiple regression analysis revealed that variation in lipid content and body weight explained 80% of the variation found in 3 fatty acids in farmed salmon, and 22:6 3 showed greater variation than other 3 fatty acids. Further analysis of lipid-corrected values revealed only minor effects of latitude on the per cent content of highly unsaturated 3 fatty acids, and hardly any effect of seawater temperature, with the exception of 22:6 3, which decreased slightly with increasing temperature.The per cent 22:6 3 in the fillet became gradually reduced with increasing fish age and body weight, whereas the content of 20:5 3 and other 3 fatty acids remained relatively constant. The per cent content of 22:6 3 of young salmon was higher than in the feed, but approached the feed value gradually as body weight increased. The lipid content of the salmon increased with fish age, and the absolute quantitative contents of both 22:6 3 and 20:5 3 increased meanwhile, even though the per cent content of 22:6 3 decreased quite pronouncedly.The per cent 22:6 3 and other 3 fatty acids was higher in wild than in farmed salmon, but the absolute quantitative content was higher throughout in farmed salmon, which had higher lipid contents. The 3/6 ratio, which is important in human health evaluation, was lower in farmed than in wild salmon. The large flexibility of 3 fatty acids and lipid content of farmed salmon leave us with the option of producing a wide variety of salmon qualities requested by the market. Both per cent and absolute quantitative 3 contents, as well as the 3/6 ratio, may readily be manipulated.     

Gonadotropins I and II do not stimulate thein vitro secretion of 17α,20β-dihydroxy-4-pregnen-3-one 20-sulphate by rainbow trout gonads during final sexual maturation

Thein vitro secretion of 17,20-dihydroxy-4-pregnen-3-one 20-sulphate (17,20-P-sulphate) and the free steroid 17,20-dihydroxy-4-pregnen-3-one (17,20-P), by rainbow trout (Oncorhynchus mykiss) gonads, in response to gonadotropin (GTH) I and GTH II, were studied during the final stages of sexual maturation. Substantial amounts of 17,20-P-sulphate were produced, by both mature ovaries and testes, indicating considerable 20-hydroxysteroid sulphotransferase (20-HST) activity within these tissues. In the post-ovulatory ovary the level of 17,20-P-sulphate (36.6 ng ml^–1) greatly exceeded that of 17,20-P (8.59 ng ml^–1). The amount of 17,20-P-sulphate produced in incubations of both mature ovary and testes was unaffected by either GTH I or GTH II treatment at physiological concentrations up to 100 ng ml^–1. Similarly, incubations of maturing ovary and testes, treated with GTH I or GTH II, in the presence of added 17,20-P at 100 ng ml^–1 of medium, produced levels of 17,20-P-sulphate that were similar to those of the controls. In incubations of mature ovarian follicles at the stages of germinal vesicle breakdown and preovulation, both GTHs significantly stimulated secretion of 17,20-P, although GTH II was always more potent than GTH I. GTH II significantly elevated the levels of 17,20-P in testicular incubations from mature males more than 4-fold relative to GTH I and controls, which did not differ from one another.In conclusion, 20-HST, the enzyme responsible for the sulphate conjugation of 17,20-P, was found to be active in the ovaries and testes of rainbow troutin vitro. However, the levels of this enzyme do not appear to be regulated by either GTH I or GTH II.     

Steroidogenesis in rainbow trout (Salmo gairdneri) at various preovulatory stages: changes in plasma hormone levels andin vivo andin vitro responses of the ovary to salmon gonadotropin

In order to specify the timing of some changes in ovarian steroid production during the transition from vitellogenesis to ovulation, plasma hormones levels andin vivo andin vitro responses of the ovary to salmon gonadotropin (s-GtH) or dibutyryl-cyclic adenosine mono-phosphate (db-cAMP) were recorded in relationship with the state of germinal vesicle migration in the oocyte.In vivo, a small, but significant, increase of plasma 17-hydroxy-20-dihydroprogesterone (17, 20-OH-P) level was detected earlier (at the subperipheral germinal vesicle stage) than the increase of GtH level (detectable at the peripheral germinal vesicle stage) and the decline of oestradiol-17 (E2–17) (also detectable at the peripheral germinal vesicle stage). Negative correlations were established between E2–17 levels and GtH (=–0.53) or 17,20-OH-P (=–0,43) levels while a positive correlation occurred between 17,20-OH-P and GtH levels (=+0,54).In vivo no action of GtH on the decline of E2–17 levels was detected GtH did not stimulate 17,20-OH-P production, within 72h, in females at the end of vitellogenesis stage. It had significant effect in females at other stages closer to ovulation, but the pattern of responses changed according to the stage.In vitro db-cAMP like GtH was able to stimulate 17,20-OH-P output from ovarian follicles. The greatest response was observed at the later stage. (GVBD). Testosterone output was also increased by GtH, but the lowest response was observed at the later stage (GVBD). Androstenedione output was lower than testosterone output.In vitro, a small but significant decline of E2–17 output was induced by GtH. We conclude that substantial changes occur during the very last stages prior to ovulation, both in the steroidogenic potential of the ovary and in the ovarian sensitivity to GtH. 20-oxydoreductase is probably progressively induced during GV migration when GtH basal levels are increasing but still relatively low. Without minimizing the role of discrete pulses of GtH on this induction, we could expect synergic actions of other hormones. Thus a high testosterone/oestradiol ratio in the follicle environment favours 17,20-OH-P secretion.     

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.     

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.     

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.     

Synthesis and regulation of 17α-hydroxy-20β-dihydroprogesterone in immature males of Oncorhynchus mykiss

Three experimental approaches were chosen to study the question if the progestin 17-hydroxy-20-dihydroprogesterone (1720OHP) is synthesised in testes of young Oncorhynchus mykiss, in which the absence of spermatozoa was verified histologically: first, in order to detect 20-hydroxysteroid dehydrogenase activity (20HSD), testes homogenates were incubated with ³H-labeled 17OHP.Metabolites were analysed by TLC, HPLC, and repeated crystallization to constant isotope ratios. One of the metabolites was identified as 1720OHP-³H, indicating that already immature testes contain 20HSD activity and are able to produce 20-reduced steroids. Second, 1720OHP was quantified by radioimmunoassay in incubates of testes fragments. The sensitivity of the gonads to gonadotropin II (GtH II) became evident when comparing incubations in the absence and presence of GtH II. Third, plasma levels of 1720OHP were significantly higher in animals injected with partially purified salmon gonadotropin, compared to controls. Thus, for the first time, it could be shown that 20HSD is present in testicular cells other than spermatozoa. Furthermore, 1720OHP is indeed secreted at a very early stage of testicular development; 1720OHP secretion is also responsive to GtH II. Future studies will have to show if the functions of this progestin include the stimulation of spermatogenesis.