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.     

Characterization of hepatic and extrahepatic glutathione S-transferases in rainbow trout (Oncorhynchus mykiss) and their induction by 3,3′,4,4′-tetrachlorobiphenyl

Liver, kidney, gill and olfactory epithelium cytosolic fractions of rainbow trout (Oncorhynchus mykiss) were examined for glutathione S-transferase (GST) contents. Proteins retained on a glutathione (GSH)-affinity matrix were separated as monomers by reversed-phase HPLC and characterized by immunoblotting, mass spectrometry and partial amino acid sequence. For each organ concerned, a specific pattern of these proteins was determined and appeared similar for liver and kidney on one hand, and for gill and olfactory epithelium on the other hand. It was confirmed that the prominent hepatic GST is a class enzyme, also constitutively expressed as a major isoform in the four organs studied. Moreover, a class variant and two new class GST subunits were characterized in minor fractions. An unknown protein, which was found major in gills and olfactory epithelium, exhibited some characteristics of class GSTs. Occurrence of possible GSH-adduct formation observed on two distinct monomers in specific experimental conditions is discussed. These results and methods were used to investigate the effect of 3,3,4,4-tetrachlorobiphenyl (TCB), a polychlorinated biphenyl (PCB), on GST expression in trout liver. From HPLC-profiling, significant co-induction of the major class and the two minor class GST subunits was observed in trout after waterborne exposure to TCB which was followed by a slight increase in 1-chloro-2,4-dinitrobenzene (CDNB) activity. The present work allows qualitative evaluation of the specific detoxification potential of rainbow trout. The use of HPLC-profiling of GSTs as a possible tool for the biomonitoring of polluted aquatic environment is suggested.     

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 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.     

Distinct alpha (α)-subunits of salmon glycoprotein hormones: production sites in the pituitary with sexual maturity

Salmon pituitary glands contain two structurally distinct -subunit proteins (1 and 2) of glycoprotein hormones: the 2-subunit is common to all salmon gonadotropins (GTH I and GTH II), whereas the 1-subunit is present in only some GTH I molecules. GTH I is predominant in the pituitary gland and plasma during gametogenesis of salmon, but the roles of the 2 GTHs in gametogenesis remain unclear. To understand the roles of GTH I, it is important to clarify patterns of 1- and 2-subunit production with sexual maturity. Thus, we produced antisera that recognized the 1- or 2-subunit, and then immunohistochemically examined the production sites of these subunits in the trout pituitary gland during ovarian development. In all pituitary glands examined, the immunoreactivity of both the 1- and 2-subunits was strong in the GTH II-producing cells, although salmon GTH II, both 1- and II-subunits, has not been detected. However, GTH I-producing cells showed a less dense immunoreactivity for 1- and 2-subunits, whereas the I-subunit was abundant. On the other hand, TSH cells, reacted with 2 but not with 1.     

Turnover of α-, γ, and δtocopherol and distribution insubcellular and lipoprotein fractions indicate presence of an hepatic tocopherolbinding protein in Atlantic salmon (Salmo salar L.)

The purpose of this work was to study the turnover of a, andtocopherol (TOH) in Atlantic salmon (Salmo salar, L.). Fish induplicate tanks were fed a diet containing 150 mg kg^-1-TOH and 100 mg kg^-1 each of and TOHadded as tocopheryl acetates. After fillet TOH concentrations had adjustedto the dietary supplementation levels, samples were taken from fish that hadbeen deprived of feed for 100 h, and from fish that had been fed regularlyuntil sampling. The retained levels of tocopherols in plasma correspondedgrossly with their biological activities, as found in experiments withmammals (::100: 20:3). The plasma concentrationsof -, and TOH amounted to 65, 44 and 15%,respectively, in unfed compared to fed fish. Very low density lipoprotein(VLDL), appeared to contain a greater fraction of plasma -TOH than ofplasma TOH. The mitochondrial fraction of liver, but not that of darkmuscle, was highly enriched in -TOH, and less in andTOH. The concentration ratios in liver and bile indicate that, and to some extent, TOH are excreted in the bile at a higherrate than -TOH. The data fit the hypothesis that Atlantic salmonliver contains a tocopherol binding protein with higher affinity for-TOH than for the other tocopherol homologues. This appears toprevent excretion of -TOH in the bile, and stimulate incorporation of-TOH in VLDL for subsequent secretion into the blood stream. As aconsequence, -TOH is retained in the body to a greater extent than and -TOH.     

Steroids and steroid glucuronides in the ovarian fluid of the African catfish,Clarias gariepinus,between ovulation and oviposition

As part of a series of experiments concerning a possible pheromonal function of steroids and steroid glucuronides excreted by the sex organs of the African catfish,Clarias gariepinus, qualitative and quantitative studies, using GCMS, were carried out to examine the presence of the steroids, that can be synthesized by the ovary during oocyte maturation and ovulation, and of the corresponding steroid glucuronides, in the fluid surrounding the eggs in the ovarian cavity shortly after ovulation.Full mass spectra were obtained of 5-pregnane-3,17-diol-20-one, 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17-triol-20-one, 5-pregnane-3,6,17,20-tetrol, 5-androstane-3,17-diol and 5-androstane-3,17-diol-11-one. After selected ion monitoring the following steroids could be detected by the presence of at least two characteristic ions at the expected retention time: 5-pregnane-3, 17,20-triol, etiocholanolone, 5-dihydrotestosterone, 5-androstane-3,11-diol-17-one, testosterone and estradiol. After treatment with -glucuronidase the following steroids could be determined in a similar way: 5-pregnane-3,17-diol-20-one, 5-pregnane-3,17,20-triol, 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17-triol-20-one, 5-pregnane,3,6,17,20-tetrol, 5-androstane-3,17-diol, etiocholanolone, 5-dihydrotestosterone, testosterone and estradiol.The free steroids 5-pregnane-3,6,17,20-tetrol and 5-pregnane-3,6,17-triol-20-one and the steroid glucuronides of testosterone, 5-dihydrotestosterone and estradiol appeared to be the most abundant of these compounds. The results indicate that very polar steroids and steroid glucuronides, synthesized in the ovary, can be excreted via the ovarian fluid shortly before and during oviposition, and possibly function as sex attractants, inducing reproductive behaviour in male conspecifics.     

Plasma profiles of fourteen ovarian steroids before,during and after ovulation in African catfish,Clarias gariepinus,determined by gas chromatography and mass spectrometry

The plasma concentrations of fourteen ovarian steroids were measured in postvitellogenic African catfish,Clarias gariepinus, which had been injected with pimozide and LHRHa. Postvitellogenesis persisted for at least four hours after pimozide and LHRHa administration. During this stage a limited rise in the plasma gonadotropin (GTH) level was accompanied by an increase in the testosterone concentration. The estradiol level was high and remained high except for a passing drop during the stage of germinal vesicle migration. At the stage of germinal vesicle migration a strong increase in the plasma GTH level coincided with a maximum in the testosterone concentration and a concomitant increase in the levels of 17,20-dihydroxy-4-pregnen-3-one and of five 5-reduced pregnanes. During germinal vesicle breakdown the GTH concentration remained high, the plasma level of 17-hydroxyprogesterone tended to increase, and the levels of 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,17,20-triol reached a maximum. At pre-ovulation the GTH concentration did not change, and peak levels were reached of 17,20-dihydroxy-4-pregnen-3-one and 5-pregnane-3,6,17-triol-20-one. Shortly after ovulation the GTH concentration slightly decreased together with a sharp decline in the concentrations of 17,20-dihydroxy-4-pregnen-3-one and the 5-reduced steroids, with the exception of 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17,20-tetrol and 5-dihydrotestosterone. The plasma concentrations of androstenedione, estrone, etiocholanolone and 5-androstane-3,17-diol showed marginal fluctuations during oocyte maturation and ovulation. Apart from 17,20-dihydroxy-4-pregnen-3-one, the 5-reduced pregnanes might be candidates for the function of oocyte maturation inducing hormone inC. gariepinus.     

The effects of epibionts and predators on the growth and mortality rates of Argopecten purpuratus cultures in southern Chile

Epibiont and predator effects on the growth and mortality of the northern scallop Argopecten purpuratus were evaluated in cultures in southern Chile.The most common epibionts were bryozoans, hydrozoans and algae in both study sites, Metri Bay (41°36; 72°43W) and Chidhuapi Channel (41°49S; 73°05W).After 12 months of culture in pearl nets and lantern nets, the scallops size did not show statistically significant differences in cultures with and without epibionts in both study sites nor were there differences in the growth either with or without the presence of predators (decapod crustaceans). The growth rate was higher in Chidhuapi Channel than in Metri Bay.Mortality was concentrated in the initial phase of culture in pearl nets. During the culture phase in lantern nets, the mortality rate was lower than 3%. The mortality rate in scallops with epibionts was higher than when these were removed in the culture phase in pearl nets. In Metri Bay the mortality rate with predators was higher than without predators. Epibionts and predators did not affect mortality in the culture phase in lantern nets.Epibionts and predation are important factors in the early mortality of scallops and therefore in the success of culture. Epibiosis, however, is not important in scallop growth in southern Chile. This is related to the composition of the epibionts and to the low temperatures which probably limit the growth of algae and invertebrates.     

Smoltification induced by a ‘skeleton’ photoperiod in underyearling coho salmon (Oncorhynchus kisutch)

Underyearling coho salmon fry were subjected to three initial photoperiod treatments (6L18D, 10L14D, 14L10D) for two months and subsequently to three final treatments (16L8D, 9L6D1L8D, 10L14D) in a factorial design. Growth rates and seawater adaptability were monitored regularly. The groups that were exposed initially to 6L18D or 10L14D and then to 16L8D grew faster and had lower plasma sodium ion levels after seawater challenge tests than any of the other groups. Fish which were initially exposed to 6 L or 10 L daylength and then to a 9L6D1L8D skeleton photoperiod, showed a slightly lower growth rate and seawater adaptability than those given the corresponding complete 16L8D photoperiod. However fish maintained on skeleton photoperiods had significantly greater growth rates and seawater adaptability than those kept on the 10L14D photoperiod. This indicates that it is not the accumulated number of hours of exposure to light that initiates smolting, but rather the time during the day when light is experienced. Fish exposed initially to 14L10D showed little or no response to subsequent changes in photoperiod, suggesting that responsiveness to inductive photoperiods depends on the initial photoperiod treatment.     

Role of thyroid hormones in tilapia larvae (Oreochromis mossambicus): II. Changes in the hormones and 5′-monodeiodinase activity during development

Thyroid hormone profiles and 5-monodeiodinase activity were determined in tilapia at different stages of early development. The results showed that both T₄ and T₃ were present in significant amounts in fertilized eggs. There was a steady decrease in both T₄ and T₃ levels during embryonic development. The levels continued to decline after hatching until around 7 days later when most of the yolk had been absorbed. The T₄ level started to rise then, suggesting that the larval thyroid had begun to produce T₄ at this time, which coincided with the period of faster growth of the larvae. The T₃ level remained fairly constant until around 20 days after which it rose significantly. In vitro determination of 5-monodeiodinase activity (5-D activity) in the whole-body homogenates of larvae showed that the enzymatic conversion of T₄ to T₃ was not detectable in eggs and 3-day-old larvae but detected in 5-day-old and older larvae. There was a gradual increase in the V_max as development proceeded indicating increasing 5-D activity during larval development. The K_m values did not differ significantly in the different stages of development. These results are discussed in relation to the growth and development of the larvae.     

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.     

Modulation of glycoprotein hormone α- and gonadotropin IIβ-subunit mRNA levels in the pituitary gland of mature male African catfish, Clarias gariepinus

The cDNAs encoding the glycoprotein hormone -subunit (GP) and the gonadotropin II-subunit (GTH II) were cloned from the pituitary gland of the African catfish, Clarias gariepinus. Using RNase protection analysis, we studied the steady-state mRNA levels of GP as well as GTH II in the pituitary gland of adult male catfish. Castration of adult male catfish resulted in a significant decrease of GTH II mRNA levels, whereas there was no change in the GP mRNA levels. Treatment of intact males with a single dose of 11-ketotestosterone (11-KT) resulted in dose-dependent increases in mRNA levels of both GP and GTH II. We conclude that 11-KT, a prominent, non-aromatizable teleost androgen, has a stimulatory effect on the pituitary mRNA levels of GP and GTH II of adult male fish.     

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.     

Steroidogenesis during induced oocyte maturation and ovulation in the African catfish,Clarias gariepinus

Changes in ovarian steroidogenesis accompanying oocyte maturation and ovulation were studied in the African catfish,Clarias gariepinus. Laboratory-reared females with postvitellogenic ovaries were treated with pimozide and LHRH-analogue. The plasma gonadotropin levels were determined by means of a homologous radioimmunoassay, the condition of the ovaries was studied by histological examination of the follicles, and the steroidogenetic capacity of the ovaries was analyzed byin vitro incubation of tissue fragments for 3 h with [³H]-pregnenolone and [³H]androstenedione as precursors. Data were collected at regular intervals between 0 and 16 h after pimozide-LHRH analogue administration.Until 4 h after the beginning of the experiments the plasma gonadotropin levels did not rise above 25 ng/ml, the ovaries remained in the stage of postvitellogenesis, and testosterone was the main end product of steroidogenesis. Four hours later the gonadotropin concentration in the blood had risen to more than 150 ng/ml, and the ovaries had entered the stage of germinal vesicle migration. At the same time steroidogenesis shifted towards the production of 17,20-dihydroxy-4-pregnen-3-one, 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,6,17-triol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,6,17,20-tetrol. During the subsequent stage of germinal vesicle breakdown the plasma gonadotropin level remained high, and the synthesis of the C₂₁-steroids showed a further increase. Simultaneously, the production of some C₁₉-steroid glucuronides was enhanced. The preovulation and especially the postovulation stages were accompanied by a gradual decrease in steroidogenic capacity of the ovaries, even though the plasma gonadotropin level remained high. It is concluded that the prematuration surge of gonadotropin influences the activity of enzymes involved in steroidogenesis, leading to a reduced C_17,20-lyase and to an augmented activity of the enzymes 20-hydroxysteroid dehydrogenase (HSD), 5-reductase, 3-HSD, 6-hydroxylase and UDP-glucuronosyltransferase. During ovulation the activity of all steroidogenic enzymes, including such key enzymes as 3-HSD and 17-hydroxylase, gradually decreases.Not only 17,20-dihydroxy-4-pregnen-3-one, but also the 5-reduced pregnanes may be involved in inducing oocyte maturation and/or ovulation. The very polar triol and tetrol products may function, together with the steroid glucuronides as sex pheromones.A preliminary account of these results was presented at the XIII Conference of European Comparative Endocrinologists, Belgrade, September 7–12, 1986     

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)

PKC activity was detected in spleen extracts from the turbot, Scophthalmus maximus, a teleost flatfish that is farmed commercially in several countries, in assays with the substrate EGF- R651–658 as phosphate acceptor. The activity was purified about 700-fold by a three-step chromatographic procedure (DEAE-cellulose, phenyl-Sepharose and threonine-Sepharose). Maximal activity was obtained in the presence of the typical PKC cofactors Ca2+ (0.1 mM) PtdS (20 g ml–1) and either DAG (2 g ml–1) or PMA (2 g ml–1). Activity was dose-dependently inhibited by H7 and by the PKC-specific inhibitors PKC19–36 and N-myristoylated PKC19–31. The rate of phosphorylation was highest with the PKC-specific substrate MARCKS161–175. In immunoblotting, MC5 (a mouse monoclonal antibody raised against bovine PKC) recognized bands of 80 and 100 kDa. Immunoblotting with antibodies raised against mouse PKC isozymes (, , , , , , and ) indicated the presence of all these isozymes in turbot spleen.     

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.     

17α,20β,21-trihydroxy-4-pregnen-3-one and 17α,20β-dihydroxy-4-pregnen-3-one stimulated spermiation in protandrous black porgy, Acanthopagrus schlegeli

The objective of this study was to investigate the effects of sex steroids on spermiation in protandrous male black porgy, Acanthopagrus schlegeli. Experiments on common carp (Cyprinus carpio) were also conducted for comparison. Fifty male black porgy were divided into 5 groups and injected with a superactive analogue of mammalian luteinizing hormone releasing hormone (LHRH-A), 17,20,21-trihydroxy-4-pregnen-3-one (20-S), 17,20-dihydroxy-4-pregnen-3-one (17,20-DHP), 11-ketotestosterone (11-KT) or saline. The dosage of the sex steroids given on days 0, 2, 4 and 6 was 330, 330, 990 and 1980 µg kg^-1 body weight, respectively. Milt volume and sperm concentrations were measured on days 0, 2, 4, 6, 8 and 10. Similar treatments were also conducted in 45 male common carp. Milt volume was significantly increased in black porgy after treatment with 20-S and 17,20-DHP; 17,20-DHP had stimulatory effects on spermiation at a lower dose (900 µg kg^-1 body weight, p < 0.05) as compared to 20-S (1980 µg kg^-1 body weight, p < 0.01). In the common carp, milt volume was also increased after treatment with LHRH-A and 17,20-DHP but not with 20-S. 17,20-DHP stimulated spermiation at a lower dose in common carp (330 µg kg^-1 body weight) than in black porgy (990 µg kg^-1 body weight). However, 11-KT did not stimulate spermiation in black porgy or common carp. The concentrations of plasma 11-KT could immediately reflect to the administration of exogenous 11-KT in black porgy.     

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.     

Blood appearance, metabolic transformation and plasma transport proteins of 14C-astaxanthin in Atlantic salmon (Salmo salar L.)

The time of appearance in blood, and transport of astaxanthin, and catabolic transformation of astaxanthin to idoxanthin were investigated in Atlantic salmon (Salmo salar) that had been force-fed a single dose of ¹⁴C-astaxanthin. In addition to the LPs, a major protein, associated with radiolabeled astaxanthin was detected. The maximum level of radiolabeled carotenoids in blood was attained 30 h after administration of ¹⁴C-astaxanthin. Radioactive idoxanthin (combined 3,4-cis and 3,4- trans glycolic isomers of idoxanthin) appeared after 6 h and a stable level was obtained after 18 h. LPDP and LP, separated by ultracentrifugation, contained on average 89 and 11% of the total radioactivity in plasma, respectively. During the 168 h experiment, maximum radioactivity in LP appeared after 22 h. Separation of plasma by ultracentrifugation on a discontinuous NaCl/KBr-gradient and an iodixanol-gradient confirmed that most of the radiolabeled carotenoids were present in the HDPF that did not contain LPs (58%), whereas HDL and LDL contained 36 and 6% of the radioactivity, respectively. Of the recovered radioactivity, astaxanthin in the HDPF comprised 82%, idoxanthin 5% and unidentified compounds 12%, whereas HDL contained 78% astaxanthin, 22% idoxanthin and no unidentified compounds. Proteins from the fractions with the high density and high radioactivity (iodixanol-gradient) were separated by PAGE under non-denaturing conditions and showed a radioactive band with parallel migration length to BSA and salmon albumin. These results show that astaxanthin is rapidly converted to idoxanthin and that the majority of astaxanthin in the plasma is associated with a protein other than LPs, presumably albumin. The identity of this protein requires verification.