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.     

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.     

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.     

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.     

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.     

Comparison of 3,5,3′-triiodo-L-thyronine and L-thyroxine absorption from the intestinal lumen of the fasted rainbow trout,Oncorhynchus mykiss

The absorptions of 3,5,3-triiodo-L-thyronine (T₃) and L-thyroxine (T₄) from the intestinal lumen of the rainbow trout were compared in vivo. Tracer doses of [¹²⁵I]T₄ (⁺T₄) or [¹²⁵I]T₃ (^*T₃) were injected through an anal cannula into the duodenum of trout fasted for 3 days at 12°C, and radioactivity was measured in blood and tissues at 4–48 h. ^*T₃ was removed more extensively than ^*T₄ from the intestinal lumen and more radioactivity was absorbed into the blood and tissues of u+T₃-injected trout than ^*T₄-injected trout. HPLC analysis showed that a high proportion of the radioactivity in the plasma, liver, kidney and intestinal lumen of ^*T₃-injected trout remained as the parent ^*T₃. However, in ^*T₄-injected trout most plasma radioactivity was in the form of ¹²⁵I^–, and by 24 h a high proportion of luminal radioactivity was ¹²⁵I^–. By 48 h, over 4% of the injected ^*T₃ and 1% of the injected ^*T₄ dose resided in the gall bladder, primarily as derivatives of ^*T₃ or ^*T₄. We conclude that T₃ is absorbed more effectively than T₄ from the intestinal lumen of fasted trout, indicating the potential for an enterohepatic T₃ cycle.     

Fatty acid compositions of the major phosphoglycerides from fish neural tissues; (n−3) and (n−6) polyunsaturated fatty acids in rainbow trout (Salmo gairdneri) and cod (Gadus morhua) brains and retinas

The fatty acid compositions of brain phosphoglycerides from a freshwater fish, the rainbow trout (Salmo gairdneri), and a marine fish, the cod (Gadus morhua), were determined and compared with those from a terrestrial mammal, the rat. Fish brain lipids were characterized by a higher degree of unsaturation encompassing increased percentages of (n–3)PUFA (226 and 205) and lower percentages of (n–6)PUFA (204 and 224). However the distribution of fatty acids and specific PUFA between different phosphoglycerides was essentially similar in rat and fish brain tissue. PE and PS contained the highest percentages of 226(n–3), PI was characterized by higher 180 and 204(n–6)/205(n–3), and PC had higher 160 and the lowest percentage of PUFA in all species. A generally similar pattern was found in the fish retinal phosphoglycerides except that PC was also rich in 226(n–3). Overall trout brain phosphoglycerides were slightly more unsaturated than the cod lipids but with lower (n–3)/(n–6) ratios whereas cod retinal lipids were more unsaturated than the trout retinal lipids.     

Catalytic activity and immunochemical quantification of hepatic cytochrome P-450 in β-naphthoflavone and isosafrol treated rainbow trout (Oncorhynchus mykiss)

In addition to catalytical assays, immunochemical techniques have recently been employed to measure induction of the cytochrome P-450 (P450) monooxygenase system in fish with polyaromatic hydrocarbons (PAH). In the present study, polyclonal antibodies were raised against rainbow trout P450IA1. Levels of rainbow trout P450IA1 determined using protein blotting- and ELISA procedures were compared with levels of 7-ethoxyresorufin-O-deethylase (7-EROD) activity in liver microsomes from rainbow trout. These comparisons showed that values of P450A1 were positively correlated (r=0.99 and r=0.97) with 7-EROD activities. In addition, the effects of isosafrol (ISF) or -naphthoflavone (NF) treatments on P450 levels in rainbow trout liver were investigated using immunochemical and catalytical methods. ISF treatment induced 7-EROD activity as well as 7-methoxycoumarin-O-demethylase-, 7-ethoxycoumarin-O-deethylase-, 7-propoxy-coumarin-O-depropylase and 7-butoxycoumarin-O-debutylase activities, although to a lesser extent, compared with the NF treatment. In contrast, immunochemical quantification of rainbow trout P450IA1 protein revealed a slightly different pattern. ISF appeared to be a weak inducer of P450IA1 in rainbow trout compared with NF. In addition, the degree of inhibition of 7-alkoxycoumarin-O-dealkylase activities in ISF microsomes differed from that measured in control- and NF microsomes. The discrepancies between catalytic and immunochemical estimates of rainbow trout P450IA1 in ISF treated fish in addition to differencs between specific inhibitory pattern by specific polyclonal antibodies raised against rainbow trout P450IA1, indicate that important differences exists between the responses induced by NF- and ISF treatments in the rainbow trout liver.Part of this work was presented at the 6th International Conference on Biochemistry and Biophysics of Cytochrome P-450, Vienna, Austria, July 3–8, 1988.     

Cortisol does not affect hepatic α- and β-adrenoceptor properties in rainbow trout (Oncorhynchus mykiss)

Distribution and function of hepatic - and -adrenoceptors were examined in rainbow trout (Oncorhynchus mykiss) injected with slow release hydrogenated coconut oil implants alone (sham) or containing cortisol. - and -Adrenoceptors were assayed on purified hepatic membranes 10–14 days post-implantation using ³H-prazosin () and ³H-CGP (). At 10–14 days, plasma cortisol values were significantly elevated to approximately 220 compared with 35.0 ng ml^-1 in cortisol implanted vs. sham trout. No significant differences were found between any of the experimental groups for either the affinity (K_d) or maximal number of binding sites (B_max) for either receptor type. Epinephrine significantly stimulated glucose release from hepatocytes isolated from sham injected trout, but not from cortisol-treated fish. Epinephrine-induced glucose release was blocked by both - and -antagonists. These studies do not support the hypothesis that rainbow trout exposed to chronic cortisol alter properties of hepatic adrenoceptors.     

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.     

Effects of acclimation temperatures of 7 and 14 °C on the contractility of Atlantic salmon (Salmo salar) intestinal smooth muscle in vitro

The contractility of intestinal smooth muscle of Atlantic salmon (Salmo salar) acclimatized to temperatures of 7 and 14 °C was compared over a prolonged period of time (9 months) in freshwater. Initial studies were carried out on intestine isolated from Atlantic salmon to estimate optimal conditions for contractility. Subsequent studies were carried out with the neurotransmitter/agonist 5-HT and TS, which activates neuronal elements in the intestine, in a Krebs-Henseleit solution containing HEPES buffer maintained at pH 7.85 and at the temperature of the experimental group (i.e., 7 or 14 °C). There was a significant interaction with time and temperature for the maximal response of 5-HT (p=0.005), effective concentration producing 50% of the maximal response (EC₅₀) to 5-HT (p=0.026) and maximal response to TS (p=0.002), demonstrating that gastrointestinal contractility of Atlantic salmon is altered by both time (month) and temperature. No significant changes were found with the effective frequency producing 50% of the maximal response (EF₅₀) to TS. The potency (EC₅₀ of 5-HT) increase between 7 and 14 °C was statistically significant (p=0.026) but the small differences found with potency were most likely not physiologically significant. A time shift was recognized in the maximal responses to 5-HT, by applying a time variable to equations for the curves of the responses. This demonstrated that the curves are similar for both temperatures but alterations in the time of the maximal response changes was different. Thus, there is a delay in the time (month) changes of gastrointestinal contractility related to acclimation temperature and would be an area for future study.     

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.     

The role of cAMP in regulating the β-adrenergic response of rainbow trout (Oncorhynchus mykiss) red blood cells

The -adrenergic response of teleost red blood cells (RBCs) enables the fish to maintain or even enhance the oxygen affinity of haemoglobin during various stress situations. The role of CAMP in the pronounced -adrenergic response of hypoxic rainbow trout RBCs was studied. Rainbow trout RBCs were incubated with three different -agonists (noradrenaline, adrenaline and isoproterenol, 10^–9 - 10^–4 M) at two oxygen tensions (P_{O 2}, 155 and 8 mmHg), and thereafter cAMP accumulation and cellular water content were measured.The cAMP concentration of non-stimulated trout RBCs was ca. 1200 nmol/kg dw. Of the three -agonists used, isoproterenol was the most effective in formation of cAMP, followed by noradrenaline and adrenaline. Oxygen tension affected the accumulation of cAMP in two ways. At physiological catecholamine levels (1–100 nM) there was either no difference between normoxic and hypoxic cells or a slight increase in the normoxic ones. At high catecholamine concentrations the accumulation of cAMP was greater in the hypoxic than in the normoxic cells. Oxygen tension also affected the magnitude of cell swelling but had no effect on the catecholamine concentrations causing half-maximal swelling (EC₅₀-values). The results indicate that, at physiological catecholamine levels, the -adrenergic response of rainbow trout RBCs is mainly regulated on the level of the Na⁺/H⁺ exchange.     

Sulfation and uptake of the maturation-inducing steroid, 17α,20β-dihydroxy-4-pregnen-3-one by rainbow trout ovarian follicles

Rainbow trout ovarian follicles were incubated in vitro with tritiated 17,20-dihydroxy-4-pregnen-3-one (17,20-P; maturation-inducing steroid). Within 18–24 h, 56–66% had been converted to tritiated 17,20-dihydroxy-4-pregnen-3-one 20-sulfate (identification confirmed by HPLC) and 27% had been taken up (absorbed) by the follicles. Addition of 125 ng of cold (non-tritiated) 17,20-P to the incubations caused a decrease in the percentage of [³H]-17,20-P which was sulfated (56% 10%) and an increase in the percentage that was taken up (27% 57%). Seven steroids were tested for their effectiveness in decreasing the sulfation and increasing the uptake of tritiated [³H]-17,20-P. The order of effectiveness was in both cases the same: 17,20-P > cortisol > 11-deoxycortisol > 17,20,21-trihydroxy-4-pregnen-3-one > 17-hydroxy-4-pregnene-3,20-dione > 17-estradiol > testosterone. This indicated that the processes of sulfation and uptake of [³H]-17,20-P were related to each other and led to the hypothesis that, when cold 17,20-P is added to the medium, it reduces the proportion of [³H]-17,20-P which is sulfated and thus allows more free [³H]-17,20-P to enter the ovarian follicles. This hypothesis was supported by the finding that each ovarian follicle had the capacity in vitro to sulfate only ca. 2 ng of [³H]-17,20-P per 18h but a capacity to take up > 500 ng per 18h.Gonadotropin I, Gonadotropin II, forskolin and phorbol-12-myristate-13-acetate (which all have an affect on steroid biosynthesis) did not affect the amount of 17,20-P which was sulfated. Sulfating activity was localized in the thecal cell layer of the follicle. The yolk fraction was shown to be responsible for absorbing the [³H]-17,20-P.     

The effects of hypoxia,in vivo,on red blood cell β-adrenoceptors in the rainbow trout,Oncorhynchus mykiss

Rainbow trout, Oncorhynchus mykiss, were exposed to 48h of environmental hypoxia (water partial pressure of oxygen = 8.0 kPa) at either 5 or 15°C. Blood was sampled during hypoxia via a dorsal aorta cannula to measure arterial blood partial pressure of oxygen and plasma catecholamine concentrations. After 48h, the number (B_max) and dissociation constant (K_d) of red blood cell surface -adrenoceptors were determined using a radioligand-displacement binding assay. At 5°C, plasma catecholamine levels were elevated at 24h whereas at 15°C, levels were elevated at 48h. At either temperature, following 48h of hypoxia, there was no change in B_max or K_d of red blood cell surface -adrenoceptors, compared to normoxic control fish. This study demonstrates that chronic exposure to moderate hypoxia does not affect the number or affinity of cell surface -adrenoceptors on trout red blood cells.     

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.     

Oxidative metabolism in a teleost,Anabas testudineus Bloch: effect of testosterone and estradiol-17β on hepatic enzyme activities

Testosterone (T) administration to maleAnabas testudineus significantly stimulated the activities of cytochrome oxidase, -glycerophosphate dehydrogenase (-GPDH) and Mg²⁺ adenosine triphosphatase (Mg²⁺ ATPase) and inhibited lactate dehydrogenase (LDH), cytosolic and mitochondrial malate dehydrogenases (MDH). The activities of succinate dehydrogenase (SDH), glucose-6-phosphate dehydrogenase (G-6-PDH) and catalase were unaffected by testosterone treatment. Administration of estradiol-17 (E2) in female fish, significantly stimulated cytochrome oxidase activity, inhibited Mg²⁺ ATPase, SDH, catalase and cytosolic and mitochondrial MDH activity, and was without effect on other enzymes studied.The simultaneous injections of actinomycin D or chloramphenicol and T or E2 prevented the hormonal influence on hepatic enzyme activities. The present study demonstrates that inA. testudineus sex steroids influence hepatic oxidative metabolism by a mechanism sensitive to the action of inhibitors of protein synthesis.     

Survival,growth and sexual maturation of the tiger trout hybrid (Salmo trutta ♀ × Salvelinus fontinalis ♂)

Brown trout ♀ × brook trout ♂ hybrids, together with parental species controls from the same breeders, were tested up to the age of 3 years at four different fish farms. Aside from egg and newborn alevin mortalities, the survival rate of the hybrids was similar to that of brown trout while that of brook trout was much lower. Hybrid growth was about intermediate between those of brown trout (inferior) and brook trout (superior). Sexual maturation was severely impaired in the hybrids, which resulted in high dressing rates: females had practically no ovarian development and male gonadogenesis, although limited, caused some morbidity during the spawning season. The feasibility of hybrid fish culture using genetic improvements is discussed in the light of these results.     

Are dietary genistein and equol potent enhancers of eicosapentaenoic acid and docosahexaenoic acid levels in rainbow trout (Oncorhynchus mykiss)?

Phytoestrogens are putatively able to enhance the biosynthesis of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), but have also been shown to affect fish growth dose dependently. The aim of the present study was to identify a concentration for the phytoestrogen genistein and the phytoestrogen metabolite equol that further increases the endogenous biosynthesis of EPA and DHA without impairing fish growth. Juvenile rainbow trout (87.2 ± 0.3 g) were fed seven diets on a fixed ratio for 8 weeks. A vegetable oil‐based diet served as a control diet and was supplemented with equol (EQ) and genistein (G), respectively, at 0.1%, 0.2% and 0.3% of feed dry matter (1, 2 and 3). Growth and nutrient composition of whole body homogenates were not affected by dietary treatments. EPA and DHA levels in liver, fillet and whole body samples were not significantly increased by EQ and G diets. Fish fed EQ diets showed dose dependently increased liver weights and C18:0 liver levels, indicating estrogen‐like effects at increased dietary dosages. In conclusion, the utilization of equol and genistein in plant oil‐based diets in order to enhance the biosynthesis of EPA and DHA seems not reasonable in rainbow trout.     

Effects of chronic exposure to γ-HCH (Lindane) on brain serotonergic and gabaergic systems,and serum cortisol and thyroxine levels of rainbow trout,Oncorhynchus mykiss

Rainbow trout (Oncorhynchus mykiss) were implanted intraperitoneally with 0.5 ml.100 g^–1 body weight of coconut oil alone (controls) or coconut oil contaning 0.05 mg of -HCH (Lindane). After 18 days, changes in brain serotonin and GABA metabolism, as well as in serum cortisol and thyroxine levels, were measured. A lower final body weight was observed in -HCH treated fish when compared with control fish. No significant differences were found for serum thyroxine levels between control and treated fish, but a significantly higher cortisol level was found in the -HCH-implanted trout. Although GABA levels did not differ significantly in any brain region in the two treatment groups, the activity of the serotonergic system was significantly altered by the pesticide in both the hypothalamus and the telencephalon.