Influence of 11-ketotestosterone, 17β-estradiol, and 3,5,3′-triiodo-L-thyronine on distribution and metabolism of carotenoids in Arctic charr, Salvelinus alpinus L.*

This investigation examines the influence of implants containing 11-ketotestosterone (11KT), 17-estradiol (E₂), and 3,5,3-triiodo-l-thyronine (T₃) on astaxanthin metabolism in sexually immature individually tagged Arctic charr. The fish (initial average weight 427 g) were maintained in freshwater for 40 days, and weekly implanted intraperitoneally with oil-based injections containing either 11 KT, E₂ or T₃ at levels of 0.1, 1.0 and 0.1 mg (100 g body weight (BW))^–1, respectively. The control fish were given the oil medium alone (0.2 ml 100 g BW^–1). The diet contained ca. 50 mg astaxanthin kg^–1. Carotenoid composition was monitored in plasma, fillet, liver and skin, and 11 KT, E₂ and testosterone (T) levels in plasma. All hormone treatments reduced plasma T compared to the control. E₂-treated fish had a higher (p<0.05) hepatosomatic index (HSI) than the other treatments. Hormone treatment did not influence gonadosomatic index (GSI). T₃ administration induced a silvery skin appearance. The fillet and plasma carotenoid content decreased during the experiment. 11 KT implantation reduced astaxanthin and idoxanthin concentrations of plasma and fillets, and increased the amount in liver and skin, compared to the other treatments. The relative proportion of astaxanthin to idoxanthin was higher in the control fish and T₃ implanted fish, than in fish implanted with 11 KT or E₂ (p<0.05). Fish treated with E₂ had the highest skin carotenoid concentration. Male fish had significantly higher carotenoid content in plasma, fillet and skin than female fish. This study reveals that sex hormones affect carotenoid metabolism and partitioning among body compartments of Arctic charr, effects differently displayed by the sexes.     

Effects of 17β-estradiol and 4-nonylphenol on smoltification and vitellogenesis in Atlantic salmon (Salmo salar)

The impact of 17-estradiol (E₂) and the putative estrogenic compound, 4-nonylphenol (4-NP), on smoltification and vitellogenesis in Atlantic salmon (Salmo salar) was investigated during a 30 day period starting late April. Three groups of fresh water (FW) fish (1 year old, mixed sexes, average weight 23 g) were injected once a week with 50 µg (0.18 µmol) 17-estradiol, 3 mg (13.6 µmol) 4-nonylphenol dissolved in peanut oil, or peanut oil alone as control. Every ten days, subgroups were challenged with 28 ppt seawater (SW) for 24h, and sampled together with subgroups of FW fish. Treatment effects were examined on vitellogenic and osmoregulatory parameters. E₂ and 4-NP treatment increased the total calcium and protein level in plasma and the hepatosomatic index of FW fish, both indicating an activated vitellogenesis in the liver. The presence of vitellogenin in the plasma of 4-NP- and E₂-treated groups was further indicated by the appearance of a high molecular weight vitellogenin band (550 kDa) in electropherograms produced by native gel electrophoresis. This band appeared in exactly the same position in both the E₂- and the 4-NP-treated groups but could not be detected in controls. During the 30 day treatment period, control fish approached the peak of smoltification, as indicated by a distinct silvery appearance, decreasing condition factor, increasing levels of gill Na⁺,K⁺-ATPase and improved hypoosmoregulatory performance in the SW-challenge test. Both E₂ and 4-NP treatments significantly inhibited the progress of smoltification, as judged by a significant reduction of gill Na⁺,K⁺-ATPase activity, relative -subunit Na⁺,K⁺-ATPase mRNA expression, gill chloride cell density and a poorer hypoosmoregulatory performance of treated fish. The impaired SW-tolerance of E₂- and 4-NP-treated fish was strongly correlated with a decreased gill Na⁺,K⁺-ATPase activity. Despite a difference in relative potency, the present study shows that 4-nonylphenol and 17-estradiol may have qualitatively similar inhibitory effects on smoltification and hypoosmoregulatory physiology of Atlantic salmon. Both 4-NP and E₂ activated the vitellogenic system, and the study supports the hypothesis that sexual maturation and smoltification are antagonistic, developmental phenomena in salmon. It is suggested that the presence of estrogenic compounds in the environment may negatively influence smoltification and migration in wild stocks of salmon.     

In vitro hepatic monodeiodination of L-thyroxine and the temporal effect of 17β-estradiol on deiodination in rainbow trout (Salmo gairdneri Richardson)

Thein vitro hepatic monodeiodination of L-thyroxine (T4) to triiodo-L-thyronine (T3) in rainbow trout (Salmo gairdneri) was found to be pH- and temperature-dependent, and was related to the amount of homogenate in the reaction vessel, suggestive of an enzyme-regulated event. Dithiothreitol (DTT) introduced into the reaction medium stimulated T3 production in a dose-related manner, whilst 6n-propyl-2-thiouracil (PTU) inhibited T3 production, also in a dose-related manner. The conversion was stimulated in the presence of light and depressed at buffer concentrations of less than 0.1 M.Prior treatment of fish with an intraperitoneal slow-release implant containing 17-estradiol (E2), at doses which are known to induce chronic mild elevations in plasma E2 levels, elicited a biphasic response to E2 as regards hepatic T3 production from T4 with a depression of T4 to T3 conversion evident within 1–2 days after implantation, and a subsequent stimulation of T3 production evident 56 days after, implantation. This increased hepatic deiodinase activity after chronic exposure to E2 at physiological doses was accompanied by a 3.5 fold increase in Vmax without a significant change in Km, suggesting the presence of an increased amount of the enzyme.     

Effects of 17β-estradiol injection on the reproductive efficiency of freshwater crayfish,Astacus leptodactylus (Eschscholtz, 1823)

The effects of 17β-estradiol injection (10^?7mol/crayfish) in adult female Astacus leptodactylus on the reproductive efficiency (i.e., pleopodal egg and stage 1 juvenile numbers) were investigated. In addition, hepatosomatic index, gonodosomatic index, and 17β-estradiol (E2) levels of crayfish ovary, hepatopancreas, and heamolymph before and after injections were also determined. Results showed that E2 injection to females enhances the reproductive efficiency of A.leptodctylus. E2 injection significantly increased pleopodal egg and stage 1 juvenile numbers, GSI, HSI, and concentrations of E2 in heamolymph, ovary, and hepatopancreas. However, E2 injection did not accelerate time of mating and proportion of pleopodal egg-berried females at the first week of reproduction season in this species. This study highlights that to improve the reproductive efficiency of A.leptodctylus by using hormones to regulate the ovarian cycle, E2 should be used at least 1.5 months before the commencement of the breeding season.     

Gender determination in the Paiche or Pirarucu (<Emphasis Type="Italic">Arapaima gigas</Emphasis>) using plasma vitellogenin, 17β-estradiol,and 11-ketotestosterone levels

Arapaima gigas is an air-breathing giant fish of Amazonian rivers. Given its great economic and cultural importance, the aquaculture development of this species represents an evident solution to face the decline of wild populations. In captivity, reproduction occurs generally in large earthen ponds where stocks of a few tens of brooders are maintained together at the beginning of the rainy season (December–March in the Peruvian Amazon). Fry production relies on the spontaneous formation of male and female pairs, which build a nest, delimit a territory and guard the offspring for at least 20 days from other congeners and predators. However, as sex determination of A. gigas is not possible by morphological criteria, it is very difficult to optimize reproduction conditions and fry production in each pond, which seriously hampers the culture of this species. This situation prompted us to develop sexing methodologies based on (1) the detection of female specific plasma Vitellogenin (Vtg) using an enzyme immuno assay (EIA), and (2) the determination of plasma 17β-estradiol and 11-ketotestosterone levels for immature specimens. The Vtg purification was performed by electro-elution after polyacrilamide gel electrophoresis (PAGE) from plasma of 17β-estradiol treated A. gigas juveniles. Two different Vtg molecules were isolated, (Vtg₁ and Vtg₂) with 184 and 112 kDa apparent molecular masses, respectively, and two antibodies were raised in rabbits for each Vtg molecule. Adult fish were 100% accurately sexed by Vtg EIA, while 100% of immature fish and 95% of adults were accurately sexed by 17β-Estradiol and 11-Ketestosterone ratios. We also observed different color pattern development in male and female adult fish (6-year-olds) around the reproductive period.     

Comparative responses in rare minnow exposed to 17β-estradiol during different life stages

Present in the excrement of humans and animals, 17β-estradiol (E₂) has been detected in the aquatic environment in a range from several nanograms to several hundred nanograms per liter. In this study, the sensitivities of rare minnows during different life stages to E₂ at environmentally relevant (5, 25, and 100 ng l⁻¹) and high (1000 ng l⁻¹) concentrations were compared using vitellogenin (VTG) and gonad development as biomarkers under semistatic conditions. After 21 days of exposure, VTG concentrations in whole-body homogenates were analyzed; the results indicated that the lowest observed effective concentration for VTG induction was 25 ng l⁻¹ E₂ in the adult stage, but 100 ng l⁻¹ E₂ in the larval and juvenile stages. After exposure in the early life stage, the larval and juvenile fish were transferred to clean water until gonad maturation. No significant difference in VTG induction was found between the exposure and control groups in the adults. However, a markedly increased proportion of females and appearance of hermaphrodism were observed in the juvenile-stage group exposed to 25 ng l⁻¹ E₂. These results showed that VTG induction in the adult stage is more sensitive than in larval and juvenile stages following exposure to E₂. The juvenile stage may be the critical period of gonad development. Sex ratio could be a sensitive biomarker indicating exposure to xenoestrogens in early-life-stage subchronic exposure tests. The results of this study provide useful information for selecting sensitive biomarkers properly in aquatic toxicology testing.     

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.     

Circulating levels and in vitro production of two maturation-inducing hormones in teleost: 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20β,21-trihydroxy-4-pregnen-3-one, in a daily spawning wrasse, Pseudolabrus japonicus

The female bambooleaf wrasse, Pseudolabrus japonicus, spawns daily during the spawning season, and exhibits a diurnal rhythm of ovarian development. In the present study, we have investigated: (1) circulating levels of 17a, 20-dihydroxy-4-pregnen- 17,20-P) and 17,20,21-trihydroxy-4-pregnen-3-one (20-S) in females sampled at different times of the day during spawning season in captivity, and (2) in vitro production of 17,20-P and 20-S by follicle-enclosed oocytes at seven different develo tal stages. In addition, we developed a microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17,20-P. Serum levels of 17,20-P and 20-S showed similar diurnal changes; substantial increases in these levels occurred around the time of germinal vesicle breakdown (GVBD). In vitro experiments showed that massive production of 17,20-P and 20-S occurred in follicles collected just before or during GVBD. Further, acute decreases in 17,20-P and 20-S production were found in the ovarian follicles just prior to ovulation, suggesting inactivation of the maturation-inducing hormone (MIH). These results, taken together with our previous data on the occurrence of GVBD in vitro, suggest a role for both 17,20-P and 20b-S as MIHs in the bambooleaf wrasse.     

Bioaccumulation of hexachlorocyclohexane,dichlorodiphenyltrichloroethane, and estradiol-17β in catfish and carp during the pre-monsoon season in India

This investigation was performed to monitor hexachlorocyclohexane isomers (HCHs), dichlorodiphenyltrichloroethane (DDT, and its metabolites, refered to as DDTs), plasma levels of estradiol-17β (E2), and the gonadosomatic index (GSI) between sampling sites of unpolluted ponds of Gujartal, Jaunpur (control site) and the polluted rivers Gomti (Jaunpur) and Ganga (Varanasi), which affect the reproductive physiology of some edible catfish and carp during the pre-monsoon season. HCHs and DDTs were measured by gas liquid chromatography (GLC) and hormones by radioimmunoassay (RIA). The results indicated that the level of HCHs and DDTs was very high in both the catfish and the carp captured from the polluted rivers compared with the fish captured from the control site. The GSI and E2 values were lower in both groups of fish when compared to the fish from the control site. The results also indicate that catfish showed greater bioaccumulation of HCHs and DDTs than carp, above the permissible limit, as compared to the fish from the control site. In conclusion, fish from the Gomti and Ganga rivers were highly polluted when compared with fish from the control site, as was evident from high levels of tissue bioaccumulation of HCHs and DDTs and decreased levels of plasma E2, inhibiting the reproductive physiology of these species at the receptor level. The levels exceeded the maximum residue limits (MRL) as recommended by Codex, hence it is suggested that the fish should be avoided for food purposes.     

The modulating effect of vitellogenin on the synthesis of 17β-estradiol by rainbow trout (Oncorhynchus mykiss) ovary

In vivo induction of vitellogenin (VTG) in response to the administration of 17-estradiol (E₂) was achieved and the protein was isolated by gel filtration column chromatography of plasma samples. Adult female trout were injected with the vitellogenic fraction every ten days from July to November and levels were measured by RIA from September to December. The results showed a significant decrease (p<0.01) in plasma E₂ levels in injected females compared with the controls. In December, after finishing the treatment, the plasma E₂ concentration increased, in injected females to reach a level similar to that of control females at vitellogenesis. The in vitro study showed that in early vitellogenic oocyte (from September) the presence of the vitellogenic fraction in the incubation medium causes a significant decrease (p<0.01) in the synthesis of E₂ by the oocytes. These data suggest that the concentration of the VTG into the oocyte can alter VTG production by the liver, moderating the production of E₂ by the ovary.     

Partial or total replacement of fish meal byplant protein affects gonadal development and plasma17 β-estradiol levels in female Nile tilapia

This study was conducted to evaluate theeffect of plant proteinbased diets on gonadaldevelopment and plasma 17 -estradiol (E2) levelin female Nile tilapia Oreochromis niloticus.Fish with a mean body weight of 6.7 (0.1) g were fedfour different diets with the same digestible protein(DP) and digestible energy (DE) containing gradedlevels of a mixture of plant ingredients as partial ortotal replacement of fish meal protein for 20 weeks.The control diet (D0) was based on fish meal, twodiets containing 33% (D33) and 66% (D66) of plantprotein, and one diet containing only plant protein(D100). Fish were sampled at 12 and 20 weeks. Nosignificant differences were found in different stagesof oocyte development and plasma E2 levels betweentilapia fed diets D0 and D100 at 12 weeks. Eight weekslater tilapia fed diet D0 showed a higher (P < 0.05)level of E2 than the D100 group. This difference andthe reduced proportion of vitellogenic and matureoocytes demonstrated that diets containing only plantprotein are less efficient in terms of tilapia growthand consequently ovarian development.     

Effect of diethylstilbestrol (DES) and 17 β-estradiol (E2) on growth, survival and histological structure of the internal organs in juvenile European catfish Silurus glanis (L.)

The effect of supplemented commercial diets with diethylstilbestrol (DES—15, 30 and 60 mg kg^?1) and 17 β-estradiol (E2—30 and 60 mg kg^?1), two chemicals commonly used in sex reversal procedure in fish, on survival and growth parameters of juvenile European catfish (Silurus glanis) was evaluated. During the two experiments, lasting 28 days each, fish were kept at temperature 25.2–26.5 °C, pH 7.4–9.3 and oxygen concentration 5.0–7.3 mg O₂ dm^?3. DES supplementation resulted in depressed growth rate of catfish. In experimental groups fed with E2, no negative effect on growth parameters was found. Both chemicals did not result in observed mortality. In all of the experimental DES groups, hepatosomatic index increased significantly, which suggests negative influence on physiological condition of catfish. DES supplementation significantly changed cytological factors of liver cells and caused hepatic alterations in parenchyma, such as vacuolization and blood congestion. Similarly, supplementation of E2 in food resulted in changes in cytological parameters of hepatocytes. However, E2 did not cause pathological changes within the liver tissue. Histological examination of the catfish gonads showed 19 and 38 % of sterile fish after treatment with 30 and 60 mg kg^?1 of DES, respectively. The results suggest that DES served in food could be ineffective in hormonal feminization process of European catfish. No disturbances of sex differentiation process after E2 treatment were observed. However, slight feminization effect in the highest level of E2 treatment group was recorded.     

Presence of maturation-promoting factor in 17α,20β-dihydroxy-4-pregnen-3-one-induced oocytes of catfish,Clarias batrachus

Full-grown immature Clarias batrachus oocytes respond in vitro to exogenous 17,20-dihydroxy-4-preg-nen-3-one ( 17,20-DP) by undergoing germinal vesicle breakdown (GVBD). Cytosolic extract (CE) prepared from 17,20-DP-induced oocytes has been shown to produce similar effect when microinjected into unstimulated immature oocytes of the same fish. A dose of 50 nl is enough to cause 100% GVBD after 4 h. Maturation-promoting factor was investigated from 17,20-DP-induced, immature and cycloheximide treated oocytes incubated in presence of [³⁵S] methionine. When the proteins were extracted and analyzed on SDS-PAGE, two prominent bands corresponding to molecular weight 34- and 46-kDa were detected in the CE of mature oocytes. However, labelling of [³⁵S] methionine was observed mainly in the region of 46 kDa protein band indicating de novo synthesis of this particular protein during l7,20-DP-induction. Further, immunoblotting study by using rabbit anti-cyclin B₁ antibody has clearly demonstrated that the protein which is newly synthesized is highly homologous to Xenopus cyclin B₁ and goldfish cyclin B.     

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.     

The dynamics of in vitro 17 β-estradiol secretion by isolated ovarian follicles of the rainbow trout (Oncorhynchus mykiss)

This study investigated the effects of human chorionic gonadotropin (hCG), 17-hydroxyprogesterone (17P) and testosterone (T) on the in vitro production of 17-estradiol (E₂) by isolated ovarian follicles of the rainbow trout (Oncorhynchus mykiss). 17P at 100 ng ml^-1, and hCG at 100 IU ml^-1 stimulated E₂ production relative to controls, whereas lower doses were ineffective. T was the most effective in stimulating E₂ production, followed by 17P and hCG respectively. The timecourse of E₂ production was investigated for both static culture, and incubations with media replacement, with follicles being exposed to hormone treatment for 30 min, 1 or 3 h, or constantly. E₂ production was observed after 30 min, 3 and 3-6 h in response to T, 17P and hCG respectively. Under static culture, E₂ levels reached maximal levels in 6 h. Longer incubations resulted in further metabolism of E₂ to E₂-glucuronide, which resulted in the blurring of treatment effects after 18 h. Incubations with media replacement resulted in higher E₂ production than in static culture. The results indicate that a 6 h incubation period is sufficient to produce significant increases in E₂ production in response to hCG, 17P and T, and that incubations longer than 12 h result in losses E₂ from the incubation media. These findings have implications for the validity of using static cultures to examine the effects of hormone treatment on the activity of steroid converting enzymes in vitro.     

Influence of chronic administration of LHRH-analogue and/or 17α-methyltestosterone on maturation in milkfish,Chanos chanos

Five chronic hormone therapies; cholesterol pellets containing 200 μg of LHRH-a (LHRH-a pellet); silastic tubing packed with either 250μg of dissolved 17α-methyltestosterone (liquid 17α-MT capsule) or 10 mg crystalline 17α-methyltestosterone (crystalline 17α-MT capsule); or the combinations of LHRH-a pellets plus a liquid 17α-MT capsule or LHRH-a pellets plus a crystalline 17α-MT capsule, were tested to determine the best treatment for inducing maturation in captive milkfish (Chanos chanos). Experimental groups of 20 milkfish each received one of these five therapies. A sixth control group received placebo implants. LHRH-a pellets were administered monthly; crystalline 17α-MT capsules were administered once, and liquid 17α-MT capsules were administered twice, at the beginning of the experiment and 3 months later.Results show that the combination of LHRH-a pellets plus liquid 17α-MT capsules in the most effective hormone therapy for enhancing the maturation of both sexes. Fifty percent of these fish matured in April, 1 month after implantation, and close to 90% of the fish in this treatment matured by July. The combination of LHRH-a pellets plus crystalline 17α-MT capsules enhanced the maturation of male milkfish but not female. LHRH-a alone was also effective in the maturation of females, but was the least effective of all treatments in maturing males. 17α-MT capsules alone, in either form, did not induce maturation in female milkfish.     

Relativein vitro effectiveness of estradiol-17β, androgens,corticosteroids, progesterone and other pregnene derivatives on germinal vesicle breakdown in oocytes of Indian major carps,Labeo rohita,Cirrhinus mrigala andCatla catla

The relative effectiveness of estradiol-17, androgens, corticosteroids, progesterone and other pregnene derivatives on germinal vesicle breakdown (GVBD) was investigatedin vitro using folliculated oocytes of three carps,Labeo rohita, Cirrhinus mrigala, andCatla catla. In all three species progesterone and 17-hydroxyprogesterone could induce GVBD but relatively 17,20-dihydroxyprogesterone was consistently found to be the most potent maturation-inducing steroid. Both estradiol-17 and testosterone were ineffective in inducing GVBD. Androsterone and dehydroepiandrosterone were found to be effective inC. catla at all the concentrations used. Deoxycorticosterone (DOC), hydrocortisone (HC) and cortisone were effective inducer of GVBD inC. catla whereas inL. rohita andC. mrigala only cortisone was found to be effective. All 5-reduced pregnenes were effective in inducing GVBD inL. rohita but inC. catla, only 5-pregnane-17-01-3,20-dione and 5-pregnane-3,17,20-triol and inC. mrigala, 5-pregnane-3-ol-20- one could induce oocyte maturation.     

Effects of estradiol-17β and 17α-methyltestosterone on gonadal differentiation in the coho salmon, Oncorhynchus kisutch

Methods are described for controlling sex differentiation in the coho salmon, Oncorhynchus kisutch. Eyed eggs and alevins were immersed in solutions of estradiol-17β and 17α-methyltestosterone and fed these steroids in the diet for a period of 10 weeks postswim-up. In seven out of 10 groups that received estradiol all fish resembled normal females when sampled at 4 months posthatch. In fish treated with methyltestosterone at all except the lowest dose (25 and 50 μg/l) the gonads resembled neither normal males nor normal females; these gonads were composed largely of connective tissue with only occasional germ cells. Various proportions of male, female and intersex gonads were observed in groups which received androgen or estrogen only in the diet. The implications of the work for salmon culture include the production of sterile fish and increasing the proportion of female salmon in hatchery populations.     

Influence of oral administration of estradiol-17β and testosterone on growth,digestion, food conversion and metabolism in the underyearling red sea bream,Chrysophrys major

Estradiol-17 (E2) administered in the diet to the red sea bream Chrysophrys major did not affect appetite, food conversion efficiency, protein efficiency ratio and specific growth rate. Serum concentrations of total protein, albumin, globulin, vitellogenin, -amino acids, total lipid, free fatty acids, cholesterol and calcium were elevated. The hepatosomatic index was also increased. Activities of hepatic enzymes including lactate dehydrogenase, isocitrate dehydrogenase and glutamate-pyruvate transaminase were higher than found in untreated control fish. Intestinal activity of leucine aminopeptidase was augmented. However, there were no changes in muscle water, protein, lipid and glycogen content. In contrast, testosterone (T) given by the same route increased appetite, food conversion efficiency, protein efficiency ratio and specific growth rate. There were no alterations in serum protein and calcium concentrations but serum glucose, ammonia and triglyceride levels were elevated. Hepatic glycogen content was increased. The activities of hepatic fructose- 1,6-diphosphatase, glucose-6-phosphatase and glycogen synthetase and intestinal activities of alkaline phosphatase and -glutamyltransferase were higher than noted on control fish. The results reveal that estradiol-17 and testosterone exerted different metabolic effects in the red sea bream and they suggest that testosterone exerts its anabolic actions by increasing appetite, food conversion efficiency and activities of digestive enzymes.