Effect of (n-3) PUFA and vitamin A Artemia enrichment on pigmentation success of turbot, Scophthalmus maximus (L).

Emulsions with different content of (n-3) PUFA and vitamin A were used to enrich Artemia and examine the effect of these nutrients on pigmentation success in turbot, Scophthalmus maximus (L.). The best pigmentation rates were obtained using an overdose of vitamin A (500 000 IU L^?1), but coincided with a high incidence of skeletal deformations. Higher growth and pigmentation rates were achieved by increasing the quantity of (n-3) PUFA oil in the emulsion. The use of (n-3) PUFA-deficient diet caused the highest occurrence of albinism as well as a cessation of metamorphosis.     

急性温度胁迫对大口黑鲈存活率及肝脏生化指标的影响

为了探究急性高温胁迫对大口黑鲈(Micropterus salmoides)“优鲈3号”存活率及肝脏生化指标的影响,将25℃(对照组)下暂养的大口黑鲈“优鲈3号”直接放入31、34、37℃水环境中,测定不同温度应激后鱼体的存活率及肝脏生化指标。结果表明:37℃组成活率低于31℃、34℃组,仅为50%。31℃和34℃组T-SOD、GSH-PX、CAT、LZM、AKP和ACP活力在48 h显著高于对照组。37℃组则与对照组无显著差异,且ACP活力在48 h显著低于对照组。结果表明,在培育大口黑鲈“优鲈3号”鱼苗时温度应该保持在34℃以下。     

Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica

Repeated injections of salmon pituitary extract (20 mg per fish per week) induced vitellogenesis in feminized, cultivated Japanese eels (Anguilla japonica). Oocytes were attained at the migratory nucleus stage after 11 or 12 injections. Addition of 17,20-dihydroxy-4-pregnen-3-one (DHP) into the incubation medium induced germinal vesicle breakdown (GVBD) in the oocytes at the migratory nucleus stage. An injection of DHP (2 µg g^-1 BW), given 24h after an injection of salmon pituitary extract (20 mg fish^-1), succeeded in inducing maturation and ovulation in females which contained occytes at the migratory nucleus stage. Most fish ovulated 15–18h following the DHP injection. Eggs that were ovulated within 15h after the DHP injection showed high fertility and hatchability, but eggs ovulated 18 or 21h after the DHP injection, showed considerably lower fertility and hatchability. A delay between ovulation and stripping of the eggs rapidly decreased both the fertility and hatchability within 6–9h after ovulation, indicating that artificial fertilization must be carried out immediately after ovulation. Repeated injections of human chorionic gonadotropin (hCG) at a concentration of 1 IU g^-1 BW week^-1 induced spermatogenesis, spermiation, and the acquisition of potential for sperm motility in cultivated males. Most males spermiated after the fifth or sixth injection of hCG, and the milt weight gradually increased and remained constant (1–2 g) from the 11th to 31th injection. Sperm motility peaked 24h after each weekly injection, and decreased from the 3^rd day after the injection. Potassium ions are an essential constituent for the maintenance of motility in the eel spermatozoa. Artificial seminal plasma containing 15.2 mM KCl is applicable as a milt diluent. Using these techniques developed for female and male eels, we have succeeded in obtaining many fertilized eggs from cultivated eels.     

Hormonal influences on in vitro [35S]-sulfate uptake by gill arches from the goldfish (Carassius auratus L.)

A bioassay for insulin-like growth factor (IGF), based on the in vitro incorporation of [³⁵S]-sulfate into gill arch tissue was used to study the hormonal regulation of proteoglycan synthesis in the goldfish (Carassius auratus). [³⁵S]-sulfate incorporation into gill arch tissue was found to be time-dependent with maximal uptake occurring by 48h, suggesting that proteoglycan synthesis in this tissue was maintained for at least 48h in vitro. The addition of human recombinant IGF-I (IGF-I) to the incubation medium was found to significantly stimulate [³⁵S]-sulfate uptake into the gill arches, whereas bovine growth hormone (GH) was without effect. Porcine insulin was also stimulatory, but results indicate that the effects of porcine insulin and IGF-I may be mediated by a single receptor system. Finally, arches from hypophysectomized fish were significantly less responsive to IGF-I than were arches from sham-operated fish. Furthermore, administration of ovine GH in vivo appeared to increase subsequent responsiveness in vitro. Together, these results provide evidence that the growth-promoting actions of GH in the goldfish may be mediated, at least in part, by a peptide related in structure to mammalian IGF-I.     

Three different levels of dietary vitamin D3 fed to first-feeding fry of Atlantic salmon (Salmo salar L.): effect on growth, mortality, calcium content and bone formation

Atlantic salmon ( Salmo salar L.) fry were reared on a fishmeal-based diet with three different levels of vitamin D₃ (0.2, 5 and 57 mg vitamin D₃ kg^–1 feed, ww) from first-feeding for 14 weeks. No significant differences were recorded in weight, length, specific growth rate, mortality, or kidney calcium concentration between the different dietary groups. No skeletal malformations or histopathological changes were recorded in any of the dietary groups. These results suggest Atlantic salmon fry to be highly tolerant of megadoses of vitamin D₃ over a limited period of time.     

草鱼体表嗜水气单胞菌拮抗细菌S190的分离鉴定及生物学特性研究

由采集的草鱼体表细菌中，分离筛选到1株抗嗜水气单胞菌较强的菌株S190。通过对该菌株个体和群体形态特征观察、生理生化特征测定及16SrDNA序列分析，初步确定为Jeotgalicoccus psychrophilus S190。杯碟法拮抗试验表明，S190对嗜水气单胞菌有较强的抑制作用；最适作用条件为25℃、pH8．0。该菌株对葡萄球菌、枯草芽孢杆菌和大肠杆菌也表现出一定程度的抑制作用。生物学特性研究表明：S190生长最适碳源为1％的糊精，氮源为0．01％的酵母粉，磷源为0．01mol／L的K2HPO4；培养温度25℃，pH10，NaCl浓度5％（m／V），接种量3％（V／V），装液量以250ml三角瓶装250ml生长最好。     

Effect of cortisol on the metabolism of 17-hydroxyprogesterone by Arctic charr and rainbow trout embryos

The effect of cortisol on the in vitro metabolism of [³H]17-hydroxyprogesterone ([³H]17OHP) was studied during embryonic development of Arctic charr (Salvelinus alpinus) and rainbow trout (Oncorhynchus mykiss). In the absence of cortisol, rainbow trout embryos metabolized [³H]17OHP largely to androstenedione (A₄) and androstenetrione (11-KA) with a minor conversion to 17,20ß-dihydroxy-4-pregnen-3-one (17,20P). In the presence of cortisol, this biosynthesis was inhibited. On the other hand, cortisol had no apparent inhibitory effect on the nature of metabolism of [³H]17OHP by Arctic charr embryos. In these embryos [³H]17OHP was metabolized mainly to 17,20P with a minor conversion to A₄ and without the formation of 11-KA that was seen in rainbow trout.When incubated in the presence of [³H]cortisol both Arctic charr and rainbow trout embryos produced 11ß-hydroxyandrostenedione (11ß-OHA) as the major metabolite, with a minor conversion to an unknown steroid. The catabolism of the cortisol by salmonid embryos may reflect the ability of the embryo to inactivate or detoxify cortisol to protect itself from the adverse effects of this biologically potent steroid hormone The study indicates the existence of species-specific differences in the nature of metabolism of [³H]17OHP and the inhibitory effect of cortisol on this metabolism.     

Alteration of liver and muscle fatty acid composition in gilthead seabream (Sparus aurata) juveniles held at high stocking density and fed an essential fatty acid deficient diet

The aim of the present study was to determine the combined effect of both stress and EFA deficiency on several biological and biochemical parameters. Fish were fed during 15 weeks two isocaloric and isoproteic diets: a control diet based on fish oil and formulated to meet the n-3 HUFA requirements for this species (1.5% of n-3 HUFA) and a deficient diet containing beef tallow and formulated to be deficient in n-3 HUFA. Each experimental diet was evaluated both at high and low stocking densities (10 and 3.2 kg m^–3 of initial density, respectively).High stocking density produced a chronic stress situation with elevation of plasma cortisol levels. It also caused a reduction in hepatosomatic index and liver lipid contents, increasing the oleic acid/n-3 HUFA ratios in the polar lipids. Fish fed the EFA deficient diet at low stocking density showed common deficiency symptoms. High stocking density in fish fed the EFA deficient diet induced a higher degree of EFA deficiency symptoms leading to mortality, liver steatosis, liver lipid deposition, reduced muscle lipid and reduced n-3 HUFA contents, which particularly affected EPA, but not DHA, suggesting a preferential retention of the latter fatty acid, specially in the phosphoglycerides fraction.     

Evidence that 17α,20β,21-trihydroxy-4-pregnen-3-one is a maturation-inducing steroid in spotted seatrout

Ovarian steroidogenesis during final oocyte maturation (FOM) in the spotted seatrout (Cynoscion nebulosus) was investigated by incubating ovarian fragments with tritiated pregnenolone, followed by chromatographic separation of the radioactive products. The major tritiated steroid produced during FOM comigrated with 17α,20β,21-trihydroxy-4-pregnen-3-one (20β-dihydro-11-deoxycortisol, 20β-S) on HPLC and TLC. Only minor amounts of radioactive material coeluted with 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-P), 11-deoxycorticosterone (DOC), estradiol-17β and testosterone standards in the HPLC system. Additional chromatography by TLC confirmed the presence of radioactive estradiol-17β and testosterone but not 17α,20β-P and DOC. All the ovarian steroids producedin vitro during FOM were assayed for their ability to induce germinal vesicle breakdown (GVBD) of spotted seatrout oocytes. Twenty grams of ovarian tissue were incubated with human chorionic gonadotropin and exogenous pregnenolone. The steroidal products were purified by HPLC and TLC. Most of the maturation-inducing activity was confined to steroidal material which comigrated in these systems with 20β-S. This material was active at a concentration of 1 ng steroid/ml medium in the GVBD assay. Smaller amounts of material which coeluted with 11-deoxycortisol, DOC, 17α,20β-P and several minor unidentified fractions induced GVBD at concentrations of 10 ng steroid(s)/ml. The structure-activity relationships of authentic steroids in inducing GVBD of spotted seatrout oocytes was investigated. Hydroxylation at the 17α, 20β or 21 positions increased potency to induce GVBD. Steroids with multiple hydroxyl groups at the 17α and 20β positions (17α, 20β-P) and at the 17α, 20β, and 21 positions (20β-S) had maximum biological activity in the GVBD bioassay. The results suggest that 20β-S is a major maturation-inducing steroid in spotted seatrout.     

Fertility and motility of sperm from Atlantic halibut (Hippoglossus hippoglossus) in relation to dose and timing of gonadotrophin-releasing hormone agonist implant

In broodstocks of Atlantic halibut, Hippoglossus hippoglossus, male and female gamete production often becomes unsynchronised towards the end of the spawning season—milt becomes very viscous and difficult to express while the females are still producing batches of good quality eggs. Gonadotrophin-releasing hormone agonist (GnRHa) has been shown to stimulate spermiation in a number of fish species. Therefore, we conducted two experiments where male halibut were implanted intramuscularly with pellets containing GnRHa. The effect of the pellets was tested at three periods: before, at the height of and at the end of spermiation. In the middle period, GnRHa was tested at two doses (5 and 25 μg/kg bodyweight). Measurements were made of milt hydration, sperm motility and fertilisation rate. Implanted males began spermiation at least 4 weeks before control males. Both doses of GnRHa increased the fluidity of the milt. This effect lasted for at least 20 days in the low dose group and for 40 days in the high dose group. When applied at the end of the season, GnRHa reversed the normal trend for the milt to become more viscous. GnRHa treatments did not affect fertilisation rates obtained with the sperm. However, towards the end of the spawning season, sperm motility was enhanced in males treated with the high dose of GnRHa (25 μg/kg) compared to controls. As described previously, plasma concentrations of the gonadal steroids, 5β-pregnane-3β,17,20β-triol 20-sulphate and 17,20-dihydroxy-4-pregnen-3-one, were significantly enhanced by GnRHa treatment. Concentrations of testosterone on the other hand decreased when spermiating males were treated with GnRHa. Our data suggest that 17,20β-dihydroxy-4-pregnen-3-one or its metabolites are involved in milt hydration, possibly through affecting ion transport.