Hypothermal effects on expression of regucalcin,a calcium-binding protein,in the livers of seawater- and fresh water–acclimated milkfish,Chanos chanos

Fish Physiology and Biochemistry - Regucalcin (RGN) is a calcium-binding protein mainly expressed in the liver. It functions in regulating activities of several calcium-dependent enzymes related to...     

Timing of early gonadal differentiation and effects of estradiol-17β treatments on the sex differentiation in Largemouth bass (Micropterus salmoides)

Fish Physiology and Biochemistry - In this study, an efficient estradiol-17β (E2)-induced feminization method was established based on the timing of early gonadal differentiation in Largemouth...     

Long-term effects of intraperitoneal injection of estradiol-17β on the growth and physiology of juvenile stellate sturgeon Acipenser stellatus

Juvenile stellate sturgeon Acipenser stellatus were intraperitoneally injected with estradiol-17β (E₂; 0 and 5 mg/kg fish) to investigate the possibility of sex reversal and also determine the changes in biochemical parameters. Five-month-old fish (40.9 ± 1.1 g) were injected every 3-week interval during a 190-day trial. At the termination of the experiment, final weight and other growth parameters including weight gain and specific growth rate, hepatosomatic and viscerosomatic indices were not affected by repetitive injection of E₂. Hematological features of E₂-treated fish showed significant reductions in number of red blood cells, hemoglobin concentration, hematocrit value and mean corpuscular hemoglobin (P < 0.05), but no significant changes were observed in number of white blood cells, mean corpuscular volume and mean corpuscular hemoglobin concentration (P > 0.05). Calcium, phosphorus, glucose, triacylglycerol, cholesterol, total protein and estradiol concentrations were significantly increased in fish injected with E₂ (P < 0.001). Plasma progesterone and testosterone levels were noticeably lower in fish injected with 5 mg/kg E₂ rather than the control fish (P < 0.001). Histological observations of gonads showed that all fish injected with 5 mg/kg E₂ apparently feminized, while 66.6 % of the control group was female. These results revealed that the injection of E₂ is an effective method for feminization of stellate sturgeon without having significant inhibitory effects on growth and survival.     

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.     

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.     

Structural and functional effects of early exposure to estradiol-17β and 17α-ethynylestradiol on the gonads of the gonochoristic teleost Dicentrarchus labrax

This study investigated the effects of estrogens on sexual differentiation in sea bass (Dicentrarchus labrax L.), a gonochoristic marine teleost that under culture conditions has a histologically sexual undifferentiated period that covers most of the first year of life, after which most individuals develop as males. Sea bass that had no noticeable histological sign of sex differentiation were fed estrogens at two doses (5 or 10 mg kg^-1 food) and for different periods ranging from 48 to 426 days post fertilization (DPF). Exposure to the synthetic estrogen 17-ethynylestradiol (EE₂) at 10 mg kg^-1 food from 60 to 260 DPF, including the sensitive period to equivalent doses of synthetic androgens previously determined for this species (126-226 DPF), significantly (p < 0.05) more than doubled the number of juvenile females to 80%, compared to the control value of 33%, and completely suppressed gonadal development in the remaining 20% of the population. This suggests that the period during which sea bass gonads exhibit high sensitivity to androgens is also very sensitive to estrogens. A comparable exposure to the natural estrogen estradiol-17 (E₂) resulted in 13% of the fish having suppressed gonadal development, but induced 57% of the fish to develop gonads with germinal tissue of both sexes, suggesting a pivotal role for E₂ during this sensitive period. Earlier exposure to EE₂ at 10 mg kg^-1 food from 48-88 DPF, significantly (p < 0.05) increased the number of females to 62% from 36% in the control group, allowing for the normal testicular development in the remaining fish. In contrast, a later chronic exposure (226-426 DPF) to E₂, at either 5 or 10 mg kg^-1 food, starting when the gonads showed no sign of sexual differentiation but past the critical sensitive period, had no effect on the resulting overall sex ratios, indicating that after this period responsiveness of the gonads to estrogens decreases as gonadal sexual differentiation progresses. However, the consequences of this apparently innocuous exposure were later manifested in adults, exemplified by a significant dose-dependent reduction in the number of mature males at 626 DPF, coinciding with the second reproductive season, the time when males normally reach sexual maturation in cultured sea bass. This suggests that chronic exposure to E₂ past the critical sensitive period may not affect the sex ratio, but could result in alterations in the male reproductive organs. This was later verified by histological analysis which revealed a significant (p < 0.05) dose-dependent reduction of the surface of the testicular lobules in the remaining males that did not mature. Together, these experiments illustrate both readily observable and subtle effects of estrogens on sex proportions, gonadal morphology and maturation rates, providing evidence that estrogen exposure can have delayed action in a teleost in a manner similar to the effects described for mammalian species. The possible existence of effects of this latter type in adult fish could be considered when evaluating the consequences of deliberate or accidental exposure to estrogens or putative estrogenic chemicals, particularly if such exposure had taken place during sex differentiation.     

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.     

Identification,characterization of selenoprotein W and its mRNA expression patterns in response to somatostatin 14, cysteamine hydrochloride, 17β-estradiol and a binary mixture of 17β-estradiol and cysteamine hydrochloride in topmouth culter (<Emphasis Type="Italic">Erythroculter ilishaeformis</Emphasis>)

In this study, a selenoprotein W cDNA was cloned from topmouth culter (Erythroculter ilishaeformis), and it was designated as EISelW. The EISelW open reading frame was composed of 261 base pairs (bp), encoding 86-amino-acid protein. The 5′ untranslated region (UTR) consisted of 104 bp, and the 3′-UTR was composed of 365 bp. A selenocysteine insertion sequence (SECIS) element was found in the 3′-UTR of EISelW mRNA. The SECIS element was classified as form II because of a small additional apical loop presented in SECIS element of EISelW mRNA. Bioinformatic approaches showed that the secondary structure of EISelW was a β1-α1-β2-β3-β4-α2 pattern from amino-terminal to carboxy-terminal. Real-time PCR analysis of EISelW mRNAs expression in 17 tissues showed that the EISelW mRNA was predominantly expressed in liver, ovary, pituitary, various regions of the brain, spinal cord and head kidney. Study of intraperitoneal injection showed that the levels of EISelW mRNA in brain, liver, ovary and spleen were regulated by somatostatin 14 (SS14), 17β-estradiol (E2), cysteamine hydrochloride (CSH) and a binary mixture of E2 and CSH, dependent on the dosage. These results suggest that E2, SS14 and CSH status may affect tissues of selenium metabolism by regulating the expression of SelW mRNA, as SelW plays a central role in selenium metabolism.     

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.     

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.     

Both TGF-β1 and Smad4 regulate type I collagen expression in the muscle of grass carp,Ctenopharyngodon idella

Fish Physiology and Biochemistry - Type I collagen is proven to make an important contribution to fish muscle quality. Our previous study has shown the Smad4-dependent regulation of type I collagen...     

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.     

Tartrate resistant acid phosphatase as a marker for scale resorption in rainbow trout,Oncorhynchus mykiss: effects of estradiol-17β treatment and refeeding

In teleosts, a considerable part of the body calcium is found in the scales. Associated with the scales are osteoblasts and osteoclasts, and during periods of high calcium demand such as during sexual maturation or starvation, the scales can be resorbed and thereby act as an internal calcium reservoir. In mammalian bone tissue, the activity of an acid phosphatase (ACP) isoenzyme, tartrate resistant acid phosphatase (TRACP), can be used as a marker for osteoclastic activity. In the present study, an evaluation of TRACP as a marker for osteoclastic activity in teleost scales has been performed. ACP and TRACP was histologically localized at resorption sites around the edge of the scales as well as at resorption holes in the scales. The optimal conditions for biochemical measurements of ACP and TRACP activity were found to be pH 5.3, 10 mM paranitrophenylphosphate, incubated for 30 min at room temperature, and 10 mM tartrate added when required. Using TRACP as a marker, estradiol-17 (E₂) was found to increase the proportion of scales being resorbed, as well as the number and size of resorption sites per scale. Also, the scales of E₂-treated fish showed weaker staining for calcium. Together, the obtained data indicate that estradiol-17 induces osteoclastic activity in teleost scales, resulting in increased resorption of the scales. A period of refeeding following a period of starvation did not have detectable effects on the scale osteoclastic activity and scale resorption.     

Studies on activity, distribution, and zymogram of protease, α-amylase, and lipase in the paddlefish Polyodon spathula

A series of biochemical determination and electrophoretic observations have been conducted to analyze the activities and characteristics of protease, α-amylase, and lipase of paddlefish Polyodon spathula. The results obtained have been compared with those of bighead carp (Aristichthys nobilis) and hybrid sturgeon (Huso dauricus ♀?×?Acipenser schrenki Brandt ♂), in order to increase available knowledge of the physiological characteristics of this sturgeon species and to gain information with regard to its nutrition. Further, a comparative study of enzymatic activity, distribution, and characterization between commercial feed-reared paddlefish (CG) and natural live food-reared (NG) paddlefish was conducted. Results showed that higher proteolytic activity was observed in the pH range 2.5-3.0 and at a pH of 7.0 for paddlefish. Levels of acid protease activity of paddlefish were similar to that of hybrid sturgeon, and significantly higher than that of bighead carp. The inhibition assay of paddlefish showed that the rate of inhibition of tosyl-phenylalanine chloromethyl ketone was approximately 2.6-fold that of tosyl-lysine chloromethyl ketone. There was no significant difference observed for acid protease activity between PG and CG groups, whereas the activity of alkaline protease, α-amylase, and lipase in the PG group were significantly lower than those in the CG group. The substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis further showed that there were certain types of enzymes, especially α-amylase, with similar molecular mass in the paddlefish and hybrid sturgeon. It can be inferred that acid digestion was main mechanism for protein hydrolysis in paddlefish, as reported for other fishes with a stomach. This indicates that the paddlefish requires higher alkaline protease, α-amylase, and lipase activity to digest natural live food.     

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.