Plasma lipoproteins in European eels (Anguilla anguilla): effects of estradiol

A dose-response effect was observed on the plasma concentration of vitellogenin (ALPP, alkali-labile protein P) over a 33 day time-course with repetitive estradiol-17 treatment of European eel (Anguilla anguilla). The plasma estradiol-17 concentration in the treated groups was not proportional to the doses injected. In the groups treated with a medium (0.5 mg kg BW^-1) or high dose (5 mg kg BW^-1) of estradiol there was a significant increase in the plasma total cholesterol concentration during the 33 day time course. The most marked dose-response effect of estradiol was that of plasma total lipids. The distribution of cholesterol between the different lipoprotein classes was dose-related: high density lipoproteins (HDL) predominated in the controls and in the low and medium dose groups, while low density lipoproteins (LDL) predominated over HDL in the group given the high estradiol dose. Plasma HDL concentration was markedly decreased in the high dose treated group; this finding is interesting since HDL plays an important role in the uptake of cholesterol from the extrahepatic tissue.     

Atlantic halibut (Hippoglossus hippoglossus) vitellogenin: induction,isolation and partial characterization

Vitellogenin (VTG) synthesis was induced by repeated injections of estradiol-17 in juvenile Atlantic halibut (Hippoglossus hippoglossus). VTG eluted as a large, phosphoprotein containing peak on DEAE-Sephacel chromatography of plasma from estradiol-17 treated juvenile and mature female, but not mature male halibut. A purification procedure for Atlantic halibut VTG was developed, where VTG was precipitated with MgCl₂, EDTA and distilled water, and the precipitated protein submitted to anion exchange chromatography on DEAE-Sephacel. Precipitated VTG eluted as a broad, partly dissociated peak on DEAE-Sephacel, when chromatography was run at 4°C, but the protein appeared intact when analysed both by SDS PAGE and native PAGE. DEAE-Sephacel chromatography at room temperature resulted in an irregular elution pattern and a dissociated protein fraction, as analysed by SDS PAGE. Biochemical characterization of VTG showed that the molecular mass of the monomer was ca 160 kDa, as estimated by SDS-PAGE. The total lipid content was 19.8% w/w, with 64%, or 12.7% of the total weight, as phospholipid. Protein bound phosphorus constituted 0.62% w/w of halibut VTG. Plasma dilution curves from mature and maturing female halibut were parallel with a dilution curve from halibut egg yolk homogenate in an homologous RIA. Plasma from mature male, but not juvenile halibut crossreacted with the VTG antiserum.     

Effects of metomidate anaesthesia or transfer to pur sea water on plasma parameters in ammonia-exposed Atlantic salmon (Salmo salar L) in sea water

Atlantic salmon (Salmo salar L) postsmolts weighing 150 ± 53 g were exposed to 14–15 mg l^–1 TA-N (total ammonia-N) in sea water in 1 m³ tanks for 24h. Blood samples were then taken A) immediately after the fish were netted from the exposure tanks and stunned by a blow to the head; B) 2–20 min after the fish were transferred to 15 l of an anaesthetic solution of metomidate in ammonia-free sea water; or C) 2–20 min after the fish were transferred to 15 l of ammonia-free sea water. Plasma TA-N level was 18% lower in the anaesthetised fish compared to in the fish sampled directly from the exposure tanks (p 0.05), and accordingly 16% lower in the fish transferred to pure sea water although this difference was not significant (p = 0.07). Plasma glucose level was higher in the fish transferred to pure sea water than in the fish receiving the two other treatments (p 0.05), but plasma urea, osmolality, Na^{+, Cl–, Ca2+ or Mg2+} levels did not vary significantly between the different treatments. Plasma TA-N level increased with time in the fish in the metomidate solution (p 0.02).     

17β-Estradiol, but not testosterone stimulates gonadotropin II concentrations in the protandrous black porgy, Acanthopagrus schlegeli Bleeker

The objective of the present study was to investigate the in vivo effects of different doses of 17-estradiol (E₂) and testosterone (T) on the levels of plasma and pituitary gonadotropin II (GTH II) in 2-year-old black porgy, Acanthopagrus schlegeli, during the spawning season. Male fish were distributed among 7 groups (n = 49), control, E₂or T (with 3 doses, 2.4 ng, 72 ng and 2.2 g g^–1 body weight). Fish were injected with the respective vehicle or different doses of E₂ or T on days 1 and 14. Plasma E₂ levels were significantly increased in the 72 ng E₂ group on days 8 and 14. Plasma vitellogenin levels were significantly higher in the 72 ng E₂ group on days 14 and 20, and 2.2 g E₂ group on days 8, 14 and 20 than those in the control group. Plasma GTH II concentrations were significantly higher in the 2.2 g E₂ group than in the control and other E₂groups on days 8, 14 and 20. Pituitary GTH II contents was significantly higher in the 7.2 ng E₂ group compared to the control and other E₂groups on day 20. Plasma GTH II concentrations were similar in the control and all the T groups on days 8, 14 and 20. None of the doses of T treatment stimulated pituitary GTH II content on day 20, although plasma vitellogenin levels were elevated. It is concluded that GTH II synthesis and secretion in black porgy is stimulated by an estrogen-specific effect.     

Characterization of growth hormone binding sites in the goldfish,Carassius auratus: effects of hypophysectomy and hormone injection

A recombinant carp growth hormone (rcGH) was used to develop for a GH radioreceptor binding assay in the goldfish (Carassius auratus). Specific binding of¹²⁵I-rcGH to goldfish liver membranes was a pH, time, temperature, and membrane protein dependent process. Scatchard and LIGAND analysis indicated a single class of high affinity and low capacity binding site, with an association constant (Ka) of 1.9×10¹⁰ M^–1 and a maximum binding capacity (B_max) of 9 fmol mg^–1 protein. Liver tissue displayed the highest¹²⁵I-rcGH binding of all the tissues examined. Displacement of¹²⁵I-rcGH with various unlabeled teleost and mammalian GHs and prolactins revealed that the goldfish hepatic binding site was highly specific for teleost GH. Intraperitoneal administration of 0.1, 1.0, and 10 g rcGH g^–1 body weight to hypophysectomized goldfish resulted in a 27, 52, and 68% decrease in total binding sites, respectively. Injection of a high dose of rat prolactin (rPRL) (5 g rPRL g^–1 body weight) also resulted in a 32% decrease in total binding sites. These results suggest that endogenous GH may have a role in the regulation of its own receptors in the goldfish.     

Gonadotropin releasing hormone-analogue treatment increases sperm motility, seminal plasma pH and sperm production in yellowtail flounder Pleuronectes ferrugineus

Compared to control fish, gonadotropin releasing hormone-analogue (GnRHa) treatment delivered either by microspheres or cholesterol pellets successfully increased sperm production (cells kg–1) and milt volume (ml kg–1) in mature yellowtail flounder Pleuronectes ferrugineus during the spawning season. Spermatocrit decreased in both treated and control groups between 12 and 29 days post-implantation, indicating a seasonal decrease in sperm concentration, rather than an effect of the GnRHa treatments.Plasma levels of testosterone, 11-ketotestoterone and 17,20dihydroxy-4-pregnen-3-one (1720P) showed no clear pattern either across treatments or over days, however this does not exclude the possiblity that GnRHa had its effect on milt volumes via the stimulation of steroid production since the sampling protocol did not allow for the rapid clearance of steroids from the plasma. GnRHa treatment did not have a negative effect on sperm fertilizing ability, percentage hatch or larval morphology. Sperm motility and seminal plasma pH were increased by GnRHa treatment.     

Cortisol treatment improves the development of hypoosmoregulatory mechanisms in the euryhaline rainbow trout,Salmo gairdneri

The effect of cortisol on osmoregulatory parameters was studied in rainbow trout, (Salmo gairdneri), kept in freshwater (FW) and/or transferred to seawater (SW). Repeated injections of 20 μg cortisol/g fish stimulated gill and gut Na⁺/K⁺-ATPase activity and reduced plasma Na⁺ and Cl⁻ levels after 2 weeks of treatment in FW-adapted fish. Cortisol doses of 0.05 and 1.0 μg/g were without effect. Repeated injections of 10 μg cortisol/g stimulated gill Na⁺/K⁺-ATPase activity and reduced plasma Na+ and Cl⁻ levels in fish in FW, and significantly improved ion regulation after their transfer to 28SW. Higher doses of cortisol (10 and 20 μg/g) induced hyperglycemia, whereas low doses (0.05 and 1.0 μg/g were without effect or induced hypoglycemia. Plasma glucose levels decreased in cortisol-treated fish transferred to SW, whereas transient hyperglycemia was seen in the control fish.     

Characterization of vitellogenin induced by estradiol-17β in protandrous black porgy,Acanthopagrus schlegeli

The objectives of this study were to isolate and characterize vitellogenin from plasma of estradiol-treated protandrous black porgy,Acanthopagrus schlegeli. Vitellogenin concentrations in plasma measured by a validated enzyme-linked immunosorbent assay (ELISA) were also compared. Two-year-old black porgy (n=20) were fed with estradiol-17 (4 mg kg^–1 of feed). Plasma was collected for purification of vitellogenin. Two forms of vitellogenin were found in plasma after chromatography on Sepharose CL-2B column and hydroxylapatite, and polyacrylamide gel electrophoresis. Black porgy vitellogenins are phospho-lipo-glycoproteins based on their chemical staining properties. The apparent molecular weights of the two forms of vitellogenin were 636 kDa and 321 kDa, respectively. The amino acid composition of purified vitellogenin was also analyzed after acid hydrolysis. The presence of immunoreactive vitellogenin was identified in the plasma and mucus extract from control and estradiol-induced females on the basis of Western blotting. Serial dilution of the plasma and mucus extract taken from estradiol-induced black porgy showed reactivity to an antiserum against lipovitellin in the ELISA, whereas mucus extract and plasma from male fish did not. Significantly higher concentrations of plasma vitellogenin were detected in estradiol-stimulated black porgy than in the control males.     

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

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

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.     

Carbohydrate utilization by juvenile silver perch, Bidyanus bidyanus (Mitchell). I. Uptake and clearance of monosaccharides following intraperitoneal injection

Abstract Intraperitoneal carbohydrate tolerance tests were done to assess the ability of silver perch, Bidyanus bidyanus, to utilize the predominant monosaccharides in plant ingredients currently being used in the formulation of aquaculture feeds for this species. Preliminary experiments carried out to assess baseline plasma glucose concentrations indicated that blood glucose levels were elevated within 2 min of handling and silver perch required a period of 48 h without feeding before plasma glucose levels remained constant. In the first carbohydrate test, either glucose, galactose or xylose were administered by injection into the intraperitoneal cavity at a dose rate of 1 g carbohydrate kg^?1 body weight (BW). In the second carbohydrate test, glucose was administered at a dose rate of either 2 or 4 g glucose kg BW^?1. Following injection, uptake and clearance rate of the carbohydrates from the blood stream was monitored over a 24‐h period. Silver perch were significantly more efficient at the uptake and clearance of glucose from the blood stream than xylose or galactose. Maximum plasma glucose concentrations (22.2 mmol L ^?1) were recorded at 1 h following injection and basal levels (3.44 mm ) were attained between 6 and 12 h following injection. For both galactose and xylose, maximum concentrations were recorded at 1 and 3 h, respectively, and concentrations of both monosaccharides remained significantly elevated 24 h after the administration. Plasma glucose concentrations of silver perch administered with either 2 or 4 g glucose kg BW^?1 were significantly elevated and peaked at similar levels (30.2 mmol L ^?1 and 30.7 mmol L ^?1 respectively) 3 h after injection. Basal plasma glucose concentrations were attained in silver perch injected with 2 g glucose kg BW^?1 at 24 h following administration. Plasma glucose concentrations remained significantly elevated in fish injected with 4 g glucose kg BW^?1 after 24 h. These findings indicate that silver perch are more efficient at utilizing glucose than either xylose or galactose, and that there are also differing maximum threshold for the inclusion of ingredients rich in glucose, galactose and xylose into the diets of silver perch.     

Development of an enzyme-linked immunosorbent assay (ELISA) for vitellogenin measurement in greenback flounder Rhombosolea tapirina

Vitellogenin (Vtg) was purified from the plasma of 17-estradiol (E₂)-injected male greenback flounder,Rhombosolea tapirina. The molecular weight of the native Vtg was estimated by gel filtration as 540 kD. SDS-PAGE and Western blotting analyses indicated that this protein consisted of three bands with molecular weights of 155, 104, 79 kD, respectively. A polyclonal antibody against the highest molecular weight band of putative Vtg was generated in sheep and an indirect antibody-capture competitive enzyme-linked immunosorbent assay (ELISA) was developed. The assay was validatedfor plasma Vtg measurement in greenback flounder. Serial dilutions of plasma from vitellogenic females parallelled the standard Vtg curve, whereas no cross-reaction was observed with the plasma of males in the ELISA. The Vtg ELISA was used to assess the induction of Vtg by E₂ in vivo in males. The induction of Vtg in greenback flounder showed a time- and dose-dependent response as in other species. In E₂-treated fish, detectable levels of Vtg were first found at 48 h, and reached a peak at 96 h post-injection. Plasma levels of Vtg increased as the E2 dose increased with a threshold of 0.1 mg kg^–1.     

Effects of aromatase inhibitors on sexual maturation in Atlantic salmon,Salmo salar,male parr

Atlantic salmon, Salmo salar, male parr were implanted with Silastic capsules filled with different aromatase inhibitors: 1,4,6-androstatriene-3,17-dione (ATD), 4-hydroxy-4-androstene-3,17-dione (4OH), and the non-steroidal CGS16949 A, 4-benzonitrile monohydrochloride (CGS). Aromatization in brain homogenates were lower in salmon implanted with CGS and ATD than in controls. This was not the case for 4OH, but administration of 4OH to brain homogenates reduced the aromatase activity. All three aromatase inhibitors had effected gonadal weights in fish sampled in the summer, but the effects were markedly different among inhibitors. Plasma levels of the androgen 11-ketotestosterone (11KT) and the progestin 17, 20-dihydroxy-4-pregnen-3-one (17,20P) were measured by means of radioimmunoassay. CGS and ATD, but not 4OH, significantly decreased the plasma 17,20P levels in the autumn. Plasma levels of 11 KT were not influenced by ATD or CGS treatment, but 4OH had a lowering effect in one autumn sampling. ATD and 4OH (CGS not tested) increased the proportion of maturing males.These findings suggest that aromatization is of physiological importance in different mechanisms controlling reproduction in salmon.     

Effect of excess iodide on thyroid function of rainbow trout,Salmo gairdneri

The acute and chronic effects of excess iodide (KI or NaI) were studied on thyroid function of rainbow trout at 11±1°C. No Wolff-Chaikoff effect, characteristic of mammals, was observed and instead plasma L-thyroxine (T₄) levels increased 6 hr after a single iodide injection. Plasma 3,5,3′-triiodo-L-thyronine (T₃) did not change and by 24 hr plasma T₄ returned to normal. This iodide-induced elevation in plasma T₄ was probably not due to toxic effects demonstrated at higher NaI or KI doses. A single iodide injection also decreased the plasma iodide distribution space, decreased the fractional rate of plasma iodide loss and completely blocked thyroidal uptake of radioiodide. Injections of iodide over a 22-day period elevated plasma iodide 200X with no mortality and no influence on plasma T₄ or T₃. It is concluded that: (i) apart from the transient 6h increase in plasma T₄, trout thyroid function, as judged by plasma hormone levels, is insensitive to considerable iodide excess, (ii) non-invasive iodide suppression of thyroidal radioiodide recycling may be useful in kinetic studies of¹²⁵I-labeled thyroid hormones, and (iii) fundamental differences in intrathyroidal iodine metabolism appear to exist between mammals and fish.     

The effects of gonadal development and sex steroids on growth hormone secretion in the male tilapia hybrid (Oreochromis niloticus × O. aureus)

Profiles of plasma growth hormone (GH) in male tilapia hybrid (Oreochromis niloticus x O. aureus) were measured and compared at different times of the year. The profiles did not appear to be repetitive, however, differences in their nature were observed at the different seasons; the most erratic profiles were seen in the height of the reproductive season (July), while the peaks were more subdued in the spring and disappeared in the autumn. Peaks in male fish were more prominent than in the females when measured in July. Perifused pituitary fragments from fish with a high GSI responded to salmon gonadotropin-releasing hormone (sGnRH) analog (10 nM-1 M), while those from fish with a low GSI barely responded to even the highest dose. Exposure of perifused pituitary fragments from sexually-regressed fish to carp growth hormone-releasing hormone (cGHRH; 0.1 M) or sGnRH (I M) stimulated GH release only after injection of the fish with methyl testosterone (MT; 3 injections of 0.4 mg kg ¹). The same MT pretreatment did not alter the response to dopamine (DA; 1 or 10 M). GH pituitary content in MT-treated fish was lower than in control fish, which may be explained by the higher circulating GH levels in these fish, but does not account for the increased response to the releasing hormones. Castration abolished the response of cultured pituitary cells to sGnRH (I fM-100 nM) without altering either their basal rate of secretion or circulating GH levels. Addition of steroids to the culture medium (MT or estradiol at 10 nM for 2 days) enabled a GH response to sGnRH stimulation in cells from sexually regressed fish. Pituitary cells which had not been exposed to steroids failed to respond to sGnRH, although their response to forskolin or TPA was similar to that of steroid-exposed cells. It would appear, therefore, that at least one of the effects of the sex steroids on the response to GnRH is exerted proximally to the formation of cAMP, or PKC, presumably at the level of the receptor. An increase in the number of receptors to the GH-releasing hormones, following steroid exposure, would explain also the changing nature of the GH secretory profile in different stages of the reproductive season.     

An homologous radioimmunoassay for brown trout (Salmo trutta) vitellogenin

An homologous radioimmunoassay for brown trout vitellogenin (VTG) was developed. Intact VTG, isolated from juvenile brown trout by selective precipitation and anion exchange chromatography was labelled with Na¹²⁵I, with 1,3,4,6-tetrachloro-3,6-diphenylglycoluril (Iodogen) as the oxidizing agent. Incorporation of Na¹²⁵I into VTG was higher than 75% and there was little degradation of the labelled protein. Labelled VTG eluted at the same position as unlabelled, purified brown trout VTG when analyzed by gel filtration on Sepharose 6B. Antisera with high titers, i.e. 1250 000, against brown trout VTG were raised in rabbits. The sensitivity of the assay was 5 ng VTG/ml and the standard curve was linear between 10 and 100 ng VTG/ml. Plasma from maturing female brown trout, as well as estradiol-treated and untreated juvenile brown trout diluted parallel to the standard curve, while plasma from maturing female rainbow trout and estradiol-treated arctic charr diluted non-parallel to the standard curve for brown trout VTG. Purified rainbow trout VTG and plasma from maturing female rainbow trout diluted parallel to each other, but with lower sensitivity than for brown trout VTG. Determinations of protein-bound phosphorus in the plasma of estradiol-treated juvenile brown trout correlated well with the RIA determinations of VTG. Repeated freezing and thawing of plasma samples yielded up to a hundred-fold increase in the apparent VTG level, while storage of a plasma sample for one year at –20°C did not affect the VTG level as measured by RIA.     

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.     

Gill Na+-K+ ATPase, carbonic anhydrase activities and plasma osmotic and ionic variations during smoltification of Atlantic salmon in the north of Portugal

Gill Na⁺-K⁺ ATPase and carbonic anhydrase activities were measured, on a fortnightly basis, from February to July, in 0₊ age Atlantic salmon (Salmo solar), hatched and reared in a freshwater experimental station in Covas, northern Portugal. Plasma osmolarity and ionic composition were also measured. Gill Na⁺-K⁺ ATPase activity increased slowly until April (15–19 moles Pi mg prot^–1 h^–1). From April to late May there was a great increase in activity (19–32 moles Pi mg prot^–1 h^–1) followed by a sharp decline in June (15 moles Pi mg prot^–1 h^–1). In contrast, carbonic anhydrase activity decreased significantly from early April to early June (170-70 moles p-nitrophenol mg prot^–1 h^–1) and increased in late June, suggesting the existence of a compensatory mechanism for the changes in Na⁺-K⁺ ATPase activity. Plasma osmolarity and sodium concentration showed lower levels during the period of high ATPase activity. On the other hand, plasma calcium concentrations showed an increase during the same period (3.47–5.98 mm1^–1 of plasma). A transitory decrease in osmolarity and plasma sodium and chlorine concentrations occurred in March, prior to the surge in Na⁺-K⁺ ATPase activity, suggesting that the physiological changes, characteristic of parr-smolt transformation can be a consequence of this loss of freshwater osmoregulatory capacity.     

Dosage-dependent differences in the effect of aromatizable and nonaromatizable androgens on the resulting phenotype of coho salmon (Oncorhynchus kisutch)

This study was conducted to investigate whether aromatization to estrogen could be the cause for the paradoxical feminization of gonads of sexually-undifferentiated fish after treatment with androgen at either high doses or for long periods. The aromatizable androgen 17-methyltestosterone (MT) and the nonaromatizable androgen 17-methyldihydrotestosterone (MDHT) were administered to groups of newly hatched coho salmon (Oncorhynchus kisutch) in a single 2h immersion at concentrations ranging from 6.25 to 6,400µg/l. The effects of treatment were evaluated by determining the resultant proportion of males in each experimental group. The effects of steroid administration on the final mean weight, length and condition factor were also determined. An increase in all these three variables was observed in the groups treated with the higher doses of MT. Regarding the resultant sexual phenotype, the response to both androgens was similar at the majority of doses tested. However, at the highest dose, the proportion of females increased with respect to that of males for MT, but not for MDHT. Since the major difference between the two androgens tested is their capacity to be aromatized, it seems that aromatization to estrogen, rather than inhibition of the biosynthesis of endogenous androgen, may explain the paradoxical feminization encountered.     

Histological changes in insulin-immunoreactive pancreatic β-cells,and suppression of insulin secretion and somatotrope activity in brook trout (Salvelinus fontinalis) maintained on reduced food intake or exposed to acidic environment

Immature brook trout (Salvelinus fontinalis) were randomly divided into a pH control, a pH and food control and an acid-stressed group. Fish in the first two groups were held at neutral pH and those in the last group were maintained at pH 4.2 for up to two months. The food supply to the pH and food control group was restricted to simulate the reduction in food intake demonstrated for acid-stressed trout. Plasma insulin levels were significantly decreased from 5–20 ng/ml to 1–2 ng/ml and plasma cortisol levels were significantly increased from 5–10 ng/ml to as high as 70 ng/ml in the acid-stressed brook trout. Concomitantly, a significant decrease of 21–39% in the proportion (volume density) of insulin immunoreactive -cells was observed within the principal pancreatic islets. Somatic growth was stunted and ultrastructural morphometry revealed the suppression of somatotrope secretory activity in the acid-stressed fish. Restriction of food supply induced a smaller but still significant decrease in circulating levels of insulin which was however not accompanied by a reduction in insulin immunoreactive -cells. The rise in plasma cortisol levels was not significant, and the plasma levels of glucose and protein were unaffected. Nevertheless, somatotrope secretory activity was suppressed and somatic growth was stunted. This study demonstrates for the first time the complexity of the endocrine response to acid stress and that some of the response to acid stress can be attributed to the lowering of food intake.