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.     

Antioxidant effect of dietary tocopherol and ascorbic acid on growth and survival of Litopenaeus vannamei postlarvae

The nutritional effect of vitamin E in dietsfor Litopenaeus vannamei postlarve (PL19)was investigated. Four formulated diets withdifferent combinations of -tocopherylacetate (-TA), ascorbic acid (AA) andhighly unsaturated fatty acids (HUFA) weretested, using four replicates.No significant differences in survival wereobserved among treatments after 34 days offeeding. However, shrimp fed with a dietcontaining 2% fish oil (low n-3 HUFA content),200 mg.kg^–1 -TA and100 mg.kg^–1 AA (diet H/E/C) showedsignificantly better growth than those fed adiet supplemented with 5% fish oil (high n-3HUFA content), 200 mg.kg^–1 -TA and100 mg.kg^–1 AA (diet H+/E/C). Shrimp fedwith a diet containing 5% fish oil,900 mg.kg^–1 -TA and100 mg.kg^–1 AA (diet H+/E+/C) showed a significantly higher tissue level of n-6 PUFAthan postlarvae fed diet H+/E/C. No definiteconclusion could be drawn about a possibleinteraction between -TA and AA, since acomparison of the diet containing 5% fish oil,200 mg.kg^–1 -TA and700 mg.kg^–1 AA (H+/E+/C+) and the dietH+/E/C did not show any significant differencesin any of the measured parameters. Theantioxidative status of the shrimp tissue(measured by means of the thiobarbituric acid(TBA) assay and expressed as nM malonaldehyde(MA) per gramme dry weight) was equal for alltreatments. Nevertheless, there was a slightlylower MA value with the diet H+/E/C+,indicating that AA may be an effectiveantioxidant in the aqueous phase and at thewater/lipid interface of the tissue. The tissuelevels of -T and AA were highlydependent on the amounts in diets and nocorrelation between -T and AAincorporation could be observed.     

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.     

Effects of Varying Dietary Fatty Acid Composition on Growth and Survival of Seahorse, Hippocampus Sp., Juveniles

Three commercially available fatty acid enrichment emulsions (DC Selco, DC DHA Selco and DC Super Selco) were used to enrich Artemia nauplii fed to seahorse, Hippocampus sp. fry. The emulsions varied in their n-3 highly unsaturated fatty acid (HUFA) composition. Total n-3 HUFA content ranged from 200 to 450mgg^-1 between the three emulsions while levels of eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) ranged between 47–220 and 80–190mgg^-1, respectively. Survival and growth of seahorses at the end of the 30 day growth trial were greater in treatments receiving enriched Artemia. Seahorses receiving Artemia enriched with DC DHA Selco and DC Super Selco showed significantly (p < 0.05) greater mean survival (71.6 ± 6.0% and 78.3 ± 6.0%, respectively) than those receiving unenriched Artemia (48.3 ± 6.0%). Mean standard length was also significantly greater (p < 0.05) in fry fed DC DHA Selco and DC Super Selco enriched Artemia (20.2 ± 0.3 and 19.7 ± 0.3mm, respectively) compared to those fed unenriched Artemia (18.1 ± 0.3mm). The results show that dietary n-3 HUFA are essential for optimal growth and survival of Hippocampus sp. and, based on the fatty acid compositions of the enriched Artemia used in this study, indicate that the level of dietary DHA supporting optimal growth and survival is greater than 9.3mgDHAg^-1 dry weight.     

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.     

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.     

Effect of expanded feed with high fish oil content on growth and fatty acid composition of rainbow trout

Rainbow trout (Oncorhynchus mykiss) were fed three different diets for 110 days — a basal dry diet with 8.4% oil content (BD8), a basal dry diet with 11.1%; oil content (BD11) a nd an expanded diet with 20.7% oil content (ED) — to investigate the influence of high fish oil exp anded diet on fatty acid composition of muscle, and to evaluate nutritional properties of edible tissue. I n fact, the experimental diets were also different in their component fatty acids, with an in creasing content of 3 highly unsaturated fatty acids (3 HUFA) from BD8 to ED. As regards biomet rics data, the condition factor and the coefficient of fatness were higher in fish fed ED in com parison with groups BD8 and BD11 (p < 0.05 ED vs. BD8). On the other hand, hepatosomatic index in group ED was markedly lower than those in groups BD8 and BD11 (p < 0.05 ED vs. BD8 and E D vs. BD11). This could be explained by the lower amount of crude protein in ED or it may indicate an excess amount of essential fatty acids (EFA) in ED. As regards fatty acid composition of fish m uscle, there were only slight differences in fatty acid composition of the edible tissue of fish wh en compared with the differences in fatty acid composition of the diets. The increased amount of fish oil in ED had a positive influence on the final weight of fish (p < 0.05 ED vs. BD8 and ED vs. BD11), but did not affect proportionately the percentage of 3 HUFA (20:53, 22:53, 22:63) and therefore the derived indices of lipid quality: so it appears possible to partially substitute fish oil in the diet with other lipid as a source of dietary fat.     

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.     

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.     

Thyroid hormones down-regulate thyrotropin β mRNA level in vivo in the turbot (Psetta maxima)

Thyroid stimulating hormone (TSH) is a vertebrate pituitary heterodimeric hormone that stimulates the thyroid gland to produce the thyroid hormones, T3 and T4. We report here the cloning, by PCR on reverse-transcribed pituitary RNAs, of a 180 bp fragment of the cDNA encoding TSH subunit in the turbot (Psetta maxima). The deduced amino acid sequence displayed 66 and 75% identity with the corresponding sequence from the European eel (Anguilla anguilla) and the rainbow trout (Oncorchyncus mykiss), respectively. This cDNA was then used as a probe for densitometric analysis of individual pituitary Northern blots. TSH mRNA levels were quantified in turbot where circulating thyroid hormones were modified by dietary treatments or hormone supplementation. Recombinant rainbow trout growth hormone had no effect on circulating thyroid hormone levels or on pituitary TSH mRNA level. In turbot fed heat-treated rapeseed meal, plasma T4 levels were lowered and TSH mRNA increased more than two fold. In contrast, when turbot were fed a standard fish-meal supplemented with T3, circulating T3 levels were elevated and there was a dramatic decrease in TSH mRNA level. It is concluded that both thyroid hormones are able to down-regulate TSH mRNA level in vivo in the turbot. These results are discussed in the context of the evolution of the TH feed-back on TSH production.     

Serum haemolytic and agglutinating activity as indicators of fish immunocompetence: their suitability in stress and dietary studies

Gilthead sea bream, Sparus aurata, were subjected to different stressful situations and the haemolytic activity mediated by the alternative complement and the agglutinating serum activity were tested as indicators of immunocompetence. The results show that both parameters were significantly compromised after chronic or repeated acute stress as well as in groups fed with vitamin E or 3 highly unsaturated fatty acids (HUFA) deficient diets and in infected fish. These results suggest that both haemolytic and agglutinating activity are suitable indicators for immunocompetence as they are non-specific responses induced by non-specific stressors. In addition, a comparison is performed with other immune indicators subjected to the same stressors.     

Effects of the aromatase inhibitor Fadrozole on plasma sex steroid secretion and oocyte maturation in female coho salmon (Oncorhynchus kisutch) during vitellogenesis

17-estradiol, 17-20-dihydroxy-4-pregnen-3-one (17-20-P), and testosterone levels were measured in plasma samples obtained from vitellogenic coho salmon (Oncorhynchus kisutch) before and 32 days after injection of the aromatase inhibitor Fadrozole (AI). Plasma 17-estradiol levels decreased significantly 6 h after injection in all AI treated fish. The higher the dose the longer the maintenance of low plasma 17-estradiol levels. Inversely, plasma 17-20-P increased significantly 6 h after injection in all AI treated fish, and the higher the dose the longer the maintenance of high plasma 17-20-P levels. At 48 h after injection plasma testosterone levels were significantly higher in the AI treated groups. The oocyte maturation index showed that multiple injections with AI retarded oocyte development. Besides, oocyte diameter and GSI were lower in the same group, which presented high incidence of atresia of vitellogenic oocytes. The ovarian follicles and brain of the fish which received multiple injections secreted less 17-estradiol, in vitro. These findings suggest that aromatase inhibitors such as Fadrozole may have a potential as a tool to regulate sexual development in salmon.     

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)     

The presence of 17α,20β-dihydroxy-4-pregnen-3-one receptor activity in the ovary of the brook trout,Salvelinus fontinalis,during terminal stages of oocyte maturation

The presence of 17,20-dihydroxy-4-pregnen-3-one (17,20-DHP) oocyte receptor activity has been demonstrated in brook troutSalvelinus fontinalis. Scatchard analyses of the cytosol fraction during various terminal stages of oocyte maturation gave a high equilibrium association constant (K_a) value of 1.394±0.669 10⁸M^–1 (n=7) and low maximum binding capacities (N_max). The association kinetics of the receptor was second order k₊₁=2.292×10⁶M^–1 sec^–1. The dissociation rate constant k_a was 1.502×10^–2 sec^–1 for the first order dissociation reaction. The K_a=1.526×10⁸M^–1, when it was determined from k₊₁/k_–1 a value close to that found from the Scatchard analysis. Competition studies showed the following binding affinities testosterone > 17-HP > 17,20-DHP > Promegestone > progesterone > estradiol > pregnenolone; cortisol showed no competitive inhibition. Cytosolic extracts when pre-equilibrated with various labelled steroids and eluted from a Sephacryl S-300 column gave multiple specific binding peaks. On sucrose density gradient centrifugation specific binding was observed at 3.05 S in cytosol containing 0.15M sodium chloride buffer. The receptor lost binding activity when incubated with various proteases, but DNase and RNase had no effect. Blood plasma without heparin at (110) dilution also bound [³H]17,20-DHP, Ka was 8.04×10⁷ M^–1.The nuclear pellet extract (750×g) gave very little specific binding activity even at high radiolabelled steroid concentrations and a linear Scatchard plot was not obtained. Nevertheless the nuclear extract, after dextran-charcoal treatment, pre-equilibrated with [³H]17,20-DHP, bound specifically to DNA cellulose, and cytosol from the same oocytes also bound to DNA cellulose under similar conditions. Although specific binding to DNA cellulose was obtained the salt concentrations at which the steroid-receptor complex elution took place was not reproducible in both nuclear extracts and cytosol samples. Also binding activity was extremely small compared to the total cytosolic binding. The nuclear extract when pre-equilibrated with high concentrations (20 nM) of the labelled steroid and then chromatographed on Sephacryl S-300 column gave a specific binding peak which was similar to that of the cytosolic preparation.The receptor levels in cytosol decreased progressively during final maturation (Stages 1–7). There is preliminary evidence for the presence of 17,20-DHP receptor activity in cytosol of landlocked Atlantic salmonSalmo salar ouananiche, and rainbow troutSalmo gairdneri. The zona radiata fraction from late stages oocyes 5, 6, and 7 in brook and rainbow trout oocytes were isolated by ultracentrifugation; from this fraction a protein was characterized which covalently bound [³H]R5020 after photoaffinity labelling. The same protein also bound [³H]17,20-DHP after solubilization in Brig 35 buffer. The SDS gel electrophoresis subunit composition of the above protein was similar to the cytosol counterpart binding [³H]17,20-DHP, although the molecular weights were different. The blood sample [³H]R5020 binding component subunit composition was different from that of the membrane extracted protein. These results demonstrate the presence of 17,20-DHP receptor activity in the cytosol and zona radiata membranes of the oocytes during final maturation.A. Maneckjee is presently NSERC postgraduate scholar at MSRL and Ph.D. candidate at Department of Biochemistry, Memorial University of Newfoundland.     

Effects of melatonin,p-chlorophenylalanine,and α-methylparatyrosine on plasma gonadotropin level and ovarian activity in the catfish,Heteropneustes fossilis: A study correlating changes in hypothalamic monoamines

The effects of (ip, 10 injections over 20 days) of melatonin (75 g 100 g^–1 BW), the serotonin (5-HT)-synthesis blocker, para-cholorophenylalanine (p-CPA, 10 mg 100g^{–1 BW}) and the catecholamine-synthesis blocker, -methylparatyrosine (-MPT, 10 mg 100 g^–1 BW) on gonadotropin (GTH) secretion and ovarian activity were studied in ^{Heteropneustes fossilis} during late preparatory to early prespawning (April–May). The treatments resulted in significant reductions of plasma GTH and estradiol-17 levels, the gonadosomatic index, frequency distribution of vitellogenic and postvitellogenic oocytes, and ovarian and serum ³²p-labelled alkali-labile phosphoprotein (a marker of vitellogenic activity). Most of the oocytes were nonvitellogenic or had undergone atretic changes. The hepatic ³²-phosphoprotein content increased significantly over the saline control value. The effects were similar and pronounced in the p-CPA and melatonin-treated groups but were moderate in the -MPT-treated group. Hypothalamic 5-HT content and turnover were significantly inhibited in the p-CPA and melatonin-treated groups but the content and turnover of catecholamines were not. The -MPT treatment decreased significantly the content and turnover of dopamine (DA), noradrenaline (NA), and adrenaline (A) but did not influence the 5-HT content or turnover. These results suggest that 5-HT, NA and A are stimulatory to GTH secretion and that melatonin may act on the serotonergic system to inhibit the pituitary-gonadal axis.to whom correspondence should be addressed.A part of the results was presented at the International Workshop on Pineal gland: Its molecular signals and published as an abstract in Neuroendocrinol. Lett. 14: 399 pp., 1992.     

Comparative composition and shelf-life of fillets of wild and cultured turbot (Scophthalmus maximus) and Atlantic halibut (Hippoglossus hippoglossus)

Turbot and Atlantic halibut are highly valued fish species. However,very little is known about fillet shelf-life characteristics associated withboth species. Thus, fillet -tocopherol content and proximate compositionof wild turbot (1.5 kg) and Atlantic halibut (1.1 kg)caught off the south coast of Ireland and the north-west coast of Iceland,respectively, were investigated. In addition, the susceptibility of fillets, storedunder retail conditions, to lipid oxidation and colour change was studied.Proximate composition analysis showed that turbot had significantly highermoisture (P < 0.001) and lower protein (P < 0.001) contents compared toAtlantic halibut. Atlantic halibut incorporated significantly higher (P <0.001) levels of -tocopherol into fillets than turbot. Over 14 days ofstorage on ice, fillets from Atlantic halibut exhibited significantly lower (P =0.020) levels of lipid oxidation than those of turbot. However, malondialdehyde(MDA) concentrations were generally very low, never exceeding 0.6 gg^–1 fillet. Turbot maintained a significantly higher (P< 0.001) pH over the storage period. The lightness (L^* values) offillets from both species increased over 14 days of storage, but wassignificantly higher (P < 0.001) in Atlantic halibut than in turbot. Turbotdeveloped a relatively intense yellow colour during storage (decrease in hueangle and increase in b^* values), whereas this was not the case forAtlantic halibut. The results of this study demonstrate that fillets of wildAtlantic halibut stored on ice, were less prone to lipid oxidation anddiscolouration than those of wild turbot. However, quality changes in turbotwere very small showing that both fish have tremendous shelf-life capacities interms of lipid oxidation. These findings are considered in the context of knownmaterial for farmed fish.     

Hormonal changes during meiotic maturation and ovulation in the brook trout (Salvelinus fontinalis)

The plasma levels of estradiol-17 (E2), 17, 20-dihydroxy-4-pregnen-3-one (17,20-P) and gonadotropin (GTH) were measured in brook trout (Salvelinus fontinalis) during the period from the end of vitellogenesis to postovulation. Blood samples were taken according to specific stages of maturation, including germinal vesicle breakdown (GVBD) and ovulation. E2 levels were quite high (45 ng/ml) at the end of vitellogenesis (and prior to GVBD) and dropped precipitously by GVBD (2 ng/ml). They remained low through ovulation and postovulation. 17,20-P levels were low prior to GVBD (0.7 ng/ml) and increased dramatically at GVBD (148 ng/ml). The levels of 17,20-P remained high at ovulation (142 ng/ml) and then dropped significantly within 24 h to approximately half of the ovulatory values. They decreased even further by 7 days postovulation. GTH levels rose gradually through GVBD and ovulation from a postvitellogenic level of approximately 3 ng/ml to a 7 day postovulatory value of approximately 10 ng/ml. The overall results; 1) decrease in estradiol prior to GVBD, 2) increase in 17,20-P at GVBD and 3) gradual GTH rise through GVBD and ovulation, are similar to those reported for other salmonids.     

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.     

Seasonal,reproductive, and nutritional influences on muscle “buffering capacity” in yellow perch (Perca flavescens)

Effective non-bicarbonate buffering capacity (or buffer value) was measured in white muscle of yellow perch (Perca flavescens) by titrations with mineral acid and base in a carbon-dioxide free, closed system. Yellow perch were collected at three month intervals throughout 1983 from an acidic lake (pH 4.6) and two alkaline lakes (pH 7.8) in northern Wisconsin. Buffering capacity was also determined for white muscle of perch kept in the laboratory under different regimes of temperature and ration. The mean buffering capacity of white muscle from yellow perch taken directly from natural environments ranged from 40.7 ± 3.1 (SD) slykes in March of 1983 to 53.7 ± 2.8 (SD) slykes in July of that year. These changes in buffering capacity were strongly correlated with water temperature. Egg production and thirty-day laboratory starvation produced significant decreases in buffering capacity and increases in the water content of yellow perch muscle. Fed perch in the laboratory had a temperature dependent buffering capacity similar to field caught fish. Buffering capacity of white muscle did not differ between yellow perch from acidic and alkaline lakes. Investigators using buffering capacity as a gauge of species differences in metabolic potential, should be wary of seasonal and reproductive factors that might alter their conclusions.     

T3 enhancement ofin vitro estradiol-17β secretion by oocytes of rainbow trout,Salmo gairdneri,is dependent on cAMP

The cellular mechanism of action of 3,5,3-triiodo-L-thyronine (T₃) in enhancing SG-G100 gonadotropin-induced ovarian secretion of 17-estradiol (E₂) was studiedin vitro using oocyte follicular preparations of rainbow trout. The dependence of the T₃ stimulatory action on the level of intracellular 3,5-cyclic adenosine monophosphate (cAMP) was shown in experiments in which forskolin or dibutyryl cAMP enhanced E₂ secretion. In the presence of partially purified salmon gonadotropin (SG-G100), T₃ stimulation of E₂ secretion was prevented by theophylline, suggesting that T₃ may exert part of its stimulatory action by inhibiting phosphodiesterase.