Effects of gonadotropins, 11-ketotestosterone,and insulin-like growth factor-1 on target gene expression and growth of previtellogenic oocytes from shortfinned eels,Anguilla australis,in vitro

Fish Physiology and Biochemistry - Pituitary gonadotropins, metabolic hormones, and sex steroids are known factors affecting the advanced stages of ovarian development in teleost fish. However, the...     

澳洲鳗鲡微卫星分子标记的筛选与检测

采用小片段克隆法构建了澳洲鳗鲡(Anguilla australis)的部分基因组文库,并用地高辛标记的(CA)15作探针筛选阳性克隆,共获得76条微卫星序列.其中10次以下CA连续重复序列占83.67%,没有检测到30次以上的连续重复序列.根据微卫星序列两端足够长的侧翼序列,设计引物55对,选择合成引物26对,用澳洲鳗鲡3个个体的混合基因组进行引物筛选,其中的18对具有清晰的扩增条带.将筛选出的18对引物对澳洲鳗鲡1个群体的40个个体进行了遗传多样性分析,其中1对引物扩增产物为单态,17对扩增产物呈现多态;17对扩增多态的引物在40个个体中扩增出等位基因数目为5～14,平均为9个.该澳洲鳗鲡群体的PIC、Ho、He的平均值分别为0.715 7、0.677 9、0.7374,所有位点均符合哈迪-温伯格平衡(P=0.05),结果证明这17个微卫星位点适于澳洲鳗鲡群体结构的研究分析.[中国水产科学,2009,16(1):133-138]     

Serum steroid profiles in artificially maturing female Japanese eel,Anguilla japonica

To investigate whether steroid profiles in salmon pituitary homogenate (SPH)-induced artificially maturing Japanese eel, Anguilla japonica, resemble those in other, naturally maturing fishes, the daily changes in 11 steroids were analyzed for a 70-day period (average time needed to reach the maturational phase). Concentrations of most steroids were low and changed on a weekly basis, with maximum values 2–5 days after an SPH injection. Thus, pregnenolone, 17α-hydroxypregnenolone, dehydroepiandrosterone, progesterone, 17α-hydroxyprogesterone, 17α,20 β-dihydroxy-4-pregnen-3-one, androstenedione and estrone levels were barely or not detectable in serum throughout the experimental period, which is largely in keeping with what is known about oogenesis-related steroids in other fishes. In contrast, serum testosterone (T) levels were high, but fluctuated considerably with each SPH injection (about 0.3–8.3 ng/ml). The serum estradiol-17β (E₂) levels increased after SPH injections and gradually rose throughout the experiment, peaking at the end of the experimental period (about 0.2–7.8 ng/ml). Serum levels of 11-ketotestosterone (11-KT) before SPH treatment were higher (approximately 2 ng/ml) than those of the other steroid hormones (less than 0.5 ng/ml). 11-KT levels increased gradually over the experimental period, and, like E₂, levels peaked towards the end of the experimental period (about 15 ng/ml). The observed patterns for T, E₂ and 11-KT are unlike those in other fishes. Furthermore, the consistent elevations in levels of 11-KT, both before and after SPH treatment, are suggestive of an important role for this steroid in controlling oocyte growth.     

Fish meal replacement by plant and animal by-products in diets for the Australian short-finned eel, Anguilla australis australis (Richardson)

In this experiment, 23% of fish meal protein was replaced with protein from Australian soybean meal (SBM), lupin meal (LM), corn gluten meal (CGM) and meat meal (MM) in nutritionally balanced experimental diets. Growth and growth efficiencies of juvenile Australian short‐finned eel Anguilla australis australis (Richardson) elvers (2.23±0.4‐g average wet weight) were compared. Elvers were fed twice a day to a total of 5% body weight per day for 63 days. Total weight gain (g), specific growth rate (% day^?1), protein efficiency ratio (%) and productive protein value (%) were significantly higher (P<0.05) for elvers fed the MM diet than for those fed the LM diet. There were no differences in diurnal ammonia‐nitrogen excretion rates. However, diurnal urea‐nitrogen excretion rates were significantly (P<0.05) affected by diet and rates were two to three times higher for all diets, except CGM, 4 h following the afternoon feed. Diet significantly affected apparent digestibility (AD); ADs of protein for SBM and LM were significantly (P<0.001) higher than for control diet, CGM and MM; AD of energy for LM was significantly (P<0.05) lower than for all the other diets. Fish meal protein was replaced by CGM, SBM and MM without compromising growth rates of the Australian short‐finned eel. However, whole lupin seed meal (LM) depressed growth and growth efficiencies most likely because of the high indigestible carbohydrate content.     

Effects of temperature on vitellogenesis in Japanese eel Anguilla japonica

ABSTRACT:   Cultured immature female Japanese eels acclimated to sea water at either 10 or 20°C were treated weekly with salmon gonadotropin fraction (sGTH) in order to investigate the effects of water temperature on artificial induction of ovarian maturation. In eels maintained at 20°C, ovulation was induced in 11 of 18 fish during the experimental period of 13 weeks, whereas at 10°C all fish showed a low gonadosomatic index (GSI) at the end of the experiment. Plasma vitellogenin levels were higher in eels kept at 20°C than in eels at 10°C throughout the experiment. However, no significant differences were observed in the plasma testosterone and estradiol-17β levels between groups. Eels pretreated with sGTHs at 10°C for 13 weeks were separated to two different temperature (10 and 20°C) groups, and received the same weekly sGTH injections. In eels transferred to 20°C, plasma vitellogenin levels, GSI and oocyte diameter were increased, but these values were maintained at low levels in eels that remained at 10°C. These results clearly indicate that water temperature is an important factor regulating vitellogenesis in the Japanese eel.     

Effects of feeding stimulants on the feed consumption,growth and survival at glass eel and elver stages in the European eel (Anguilla anguilla)

Elvers and glass eels of Anguilla anguilla were fed diets containing two types of feeding stimulants (FS) that were based on processed marine (MBFS) and yeast proteins (YBFS). Elvers (1.5 ± 0.3 g) were fed seven diets (MBFS and YBFS diets at 20 g kg^?1, 40 g kg^?1 and 60 g kg^?1 plus control) for 60 days. Glass eels (250 ± 100 mg) were weaned to 60 g kg^?1 MBFS, 60 g kg^?1 YBFS and control diets for 30 days. Diets containing 60 g kg^?1 FS had a beneficial effect in terms of growth, homogenous size distribution and feed intake in elvers. Elvers fed 60 g kg^?1 MBFS and YBFS diets grew 11.9% and 5.6% faster than the control group. No differences in growth and size distribution were detected in glass eels fed 60 g kg^?1 MBFS and YBFS diets. However, FS affected the digestive system maturation; fish fed the 60 g kg^?1 MBFS and YBFS diets showed higher and intermediate values in the specific enzyme activities in comparison with the control group. This study revealed that the incorporation of FS into a pelleted diet was beneficial on the overall performance of European glass eels and elvers. However, the observed results were different depending on the eel’s stage of development, as well as the type and inclusion level of the FS.     

Preliminary observations on the tank and pond culture of the glass eels of the Australian shortfin eel, Anguilla australis Richardson

There is expanding interest in the culture of the Australian shortfin eel Anguilla australis Richardson; however, there is a lack of fundamental biology and husbandry information necessary to further develop an industry within Australia. The present study was undertaken to gain a preliminary understanding of basic husbandry requirements for rearing of juvenile A. australis (glass eels and elvers) in tanks and earthen ponds. Newly caught glass eels were successfully acclimated to culture conditions. During tank culture trials, specific growth rates (SGR) and survival rates ranged from ?2.1 to 2.8% day^?1 and 52% to 100% respectively. Glass eels weaned onto a commercial eel diet exhibited a significantly greater SGR and survival rate than those weaned onto a commercial trout diet. Glass eels weaned onto an eel diet over a 15‐day period grew slightly faster than eels weaned over a 5‐day period, but survival rates were not significantly different for each treatment. SGRs (up to 2.8% day^?1) were significantly higher for glass eels fed at 9 and 12% day^?1 than at 6% day^?1. Stocking densities between 2.5 kg m^?3 and 30 kg m^?3 did not influence either SGR or survival rates. SGRs were significantly higher for glass eels cultured at 25 °C than at lower temperatures. During pond culture trials, SGRs and survival rates ranged from 1.36 to 1.65% day^?1 and 39% to 77% respectively. The SGR and survival rates of juvenile eels stocked into ponds receiving supplementary feeding with a commercial eel diet were not significantly different to those of eels stocked into ponds that did not receive supplementary feeding.     

Effects of ambient ion concentrations on gill ATPases in fresh water eel,Anguilla anguilla

Branchial activities of Na⁺,K⁺-ATPase (ouabain sensitive), Mg²⁺ ATPase (ouabain insensitive) and kinetic analysis of high and low affinity Ca²⁺ ATPase were measured inAnguilla anguilla that had been acclimated to demineralized water (DW, Ca < 10 M), freshwater (FW, Ca = 2 mM), and Low calcium freshwater (L-Ca, Ca = 0.9 mM). Na⁺,K⁺-ATPase activity decreased while ouabain insensitive activity increased when ambient Ca²⁺ decreased. Two kinetic forms of Ca²⁺ ATPase could be resolved in each environmental condition. The stimulation coefficients of both sites or enzymes were not affected by ambient Ca²⁺ concentrations. The maximal velocity of both the high and the low affinity Ca²⁺ ATPase was increased when external Ca²⁺ was decreased during acclimation. The low affinity Ca²⁺ ATPase and the Mg²⁺ stimulated enzyme could be a non specific enzyme accepting either Ca²⁺ or Mg²⁺. Results are compared with previous results in the literature and in relation to the branchial morphology and ionic exchanges in fish.     

Growth of Australian shortfin eel (Anguilla australis) elvers given different dietary protein and lipid levels

The Australian shortfin eel, Anguilla australis is a potential candidate for intensive aquaculture. The present study was undertaken to evaluate the growth of elvers (5.4 g ± 0.1 initial weight) fed with diets of varying protein and lipid content, and to assess the potential of using soya‐bean meal as a dietary ingredient. A 10 week experiment was conducted at 24 (±1.0) °C by rearing fish, in 60 L conical fibre glass tanks using a closed recirculation system. Diets having protein concentrations of 40 or 50% (by dry weight) were tested at three lipid levels (15, 20, 25%); diets being designated P₄₀L₁₅, P₄₀L₂₀, P₄₀L₂₅, P₅₀L_15, etc. All these diets contained 5% soya‐bean meal. In addition P₅₀L₂₀ diets were formulated to contain 10 and 20% soya‐bean meal in the diet (Diets S1 & S2). Shortfin eel grew best on the P₅₀L₁₅ diet, with an average specific growth rate of 2.26%. Food conservation ratio (FCR) and Protein efficiency ratio (PER) ranged from 1.21 (P₅₀L₁₅) to 2.12 (P₄₀L₂₅), and 0.92 (P₅₀L₂₅) to 1.65 (P₅₀L₁₅), respectively. Based on all criteria the best growth performance of shortfin eel was on the P₅₀L₁₅ diet, followed by P₄₀L₂₀ and P₄₀L₁₅. At both protein levels fish reared on diets with 25% lipid performed poorly. The performance of shortfin eel was not affected by the amount of soya‐bean meal in the diet, up to a maximum of 20% dietary inclusion. No significant differences in muscle protein were evident in shortfin eel reared on different dietary treatments, nor was the lipid content of muscle related to dietary lipid level.     

Effects of ammonia accumulation on metabolic rates and growth of European eel, Anguilla anguilla L., in relation to warmwater aquaculture

Abstract. Laboratory experiments on fingerling eels, Anguilla anguilla L., were carried out at high density (50gl¹) in fresh and sea water (30% salinity) at 25°C. The effects of reduced water exchange rates, leading to excretory ammonia accumulation, on feeding, assimilation, oxygen consumption, ambient pH and nitrogen excretion were studied. Estimates of growth rates were made. High ambient ammonia concentrations caused large and prolonged increases in post-feeding oxygen consumption, energy wastage and growth suppression. The main causes are suggested to be hyperventilation and neural hyperexcitability due to increases in blood unionized ammonia. The data obtained are used to predict minimum water exchange rates needed to prevent ammonia-induced growth suppression and to meet peak post-feeding oxygen demands.     

Effects of dietary propolis supplementation on growth performance, immune responses, disease resistance and body composition of juvenile eel, Anguilla japonica

This study was conducted to investigate the effects of dietary propolis supplementation on growth performance, immune responses, disease resistance and body composition of juvenile eel, Anguilla japonica, in order to evaluate its bioavailability as a feed additive for this species. A total of 540 fish averaging 7.7?±?0.22?g (mean?±?SD) were randomly distributed into 18 tanks in groups of 30, and each tank was then randomly assigned to one of three replicates of six diets containing 0 (P₀), 0.25 (P_0.25), 0.5 (P_0.5), 1.0 (P₁), 2.0 (P₂) and 4.0 (P₄) % dietary propolis. At the end of 12?weeks of feeding trial, weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) of fish fed P_0.5 diet were significantly higher than those of fish fed P₀, P₁, P₂ and P₄ diets (P?<?0.05). These parameters were 148.9%, 0.72% day^?1, 94.4% and 2.9, respectively for fish fed P_0.5 diet. Serum lysozyme activity of fish fed P_0.5 (105.7 units mL^?1) and P₁ (106.0 units mL^?1) diets were significantly higher than those of fish fed P₀, P_0.25, P₂ and P₄ diets. Mucus lysozyme activity of fish fed P₁ (8.4 units 10?cm^?2) diet was significantly higher than those of fish fed P₀, P₂ and P₄ diets. Results indicated that the optimum dietary propolis supplementation levels could be 0.25?C0.5% for optimum growth and feed efficiency, and 0.5?C1% for enhanced immune responses and disease resistance in eel, A. japonica. This study may suggest that the dietary propolis level for the optimum immune responses could be higher than the level for the optimum growth of eel.     

鳗鲡拟指环虫的免疫原性

拟指环虫病是严重危害养殖鳗鲡(Anguilla sp.)的寄生虫病,为了寻求更为安全有效的免疫防治方法,对鳗鲡拟指环虫(Pseudodactylogyrus spp.)的结构蛋白及其免疫原性进行分析。以鳗鲡拟指环虫全虫蛋白为抗原,制备了鼠和鳗鲡抗拟指环虫免疫血清,ELISA检测效价分别为1∶51 200和1∶3 200。SDS-PAGE分析表明,虫体含有16 kD、21 kD、29 kD、37 kD、43.5 kD6、8 kD和110 kD等主要结构蛋白,其中29 kD的多肽为高丰度蛋白;Western blot证实免疫鳗和康复鳗血清均能识别25 kD、43.5 kD6、2 kD、81 kD等虫体蛋白。间接荧光抗体试验显示,个别虫体在头部、尾部有明显的荧光染色,大多数虫体中部两侧,特别是卵黄腺丰富区域,呈较强的荧光染色。     

Effects of attractants on start of feeding of glass eel, Anguilla anguilla L.

Abstract. In four experiments, performed at the Agricultural University, Wageningen and the Netherlands Institute for Fishery Investigations, the effects of attractants on feed acceptance and growth of glass eels, Anguilla anguilla L., were studied. The results show that extracts from natural food or a mixture of synthetic amino acids improve acceptance of a trout fry crumble at first feeding of glass eels. Overall results can be greatly improved by initiating the feeding behaviour of the fish by feeding cod roe for the first few days. The results also demonstrate a strong effect of the feeding level on feed acceptance. At relatively high feeding levels no significant effect of attractants could be detected.     

Functional partition of cells in the gill epithelium of the Japanese eel, Anguilla japonica, and the role of hormones

Viable chloride and pavement cells were isolated from the gill epithelium of Japanese eel, Anguilla japonica, by a 3-step percoll gradient centrifugation at low speed. Viability of the isolated cells were tested by the trypan blue exclusion test, rhodamine-123, or pre-labelling the cells with fluo-3 Ca²⁺ dye and examined by laser confocal microscopy. Isolated chloride cells responded to ionomycin with a rapid increase in Ca²⁺ fluorescence, which was abolished by chelating external Ca²⁺ with EGTA. Peptide hormones, including arginine vasotocin, isotocin, insulin-like growth factor I and II, and urotensin I increased Ca²⁺ entry, urotensin II had no effect, and eel corpuscles of Stannius extract reduced residual Ca²⁺ fluorescence. Isolated chloride cells and pavement cells from eels were analyzed for their enzymatic activities involved in intermediary and nitrogen metabolism. Chloride cells had high levels of glutaminase I, glutamate dehydrogenase, glutamate oxalacetate transaminase, HCO₃-ATPase and carbamoyl phosphate synthetase II. Pavement cells had highly active glucose-6-phosphate dehydrogenase and AMP deaminase. Both had high levels of lactate dehydrogenase compared with other tissues.     

Effects of dietary carbohydrate source on growth and body composition of the European eel (Anguilla anguilla L.)

Different carbohydrate sources—wheat meal, bread meal, soluble corn starch, native potato starch and sorghum meal—were included at 30% in isonitrogenous diets for European eels (Anguilla anguilla) weighing 4.6 g kept in tanks. Growth and body composition were determined and feed utilization was evaluated. Total gains/m² per day were as follows: on wheat, 3.20 g; bread meal, 2.77 g, soluble starch, 2.28 g; sorghum, 1.21 g; and potato, 0.78 g.Percent of protein in whole carcasses was similar for all diets, but percent of fat was higher with wheat and bread meal and lowest with potato starch. Food conversion, protein retention and energy retention were higher for eels fed wheat meal or bread meal than the other carbohydrates.     

日本鳗鲡产卵后生存及其繁育

为证明日本鳗鲡(Anguilla japonica)生活史最后一步一产后鳗的命运,本研究模拟产后的日本鳗鲡继续在海水中养殖,观察其存活率及繁育情况.结果表明,产后鳗在海水中停食约18 d后,体能得到恢复,部分亲鱼开始出现摄食,1个月左右全部恢复摄食,经244 d养殖,雌、雄鳗体质量增加,存活率达94.6%.随后,给产后鳗注射外源性促性腺激素(鲤鱼脑垂体匀浆CPE和人绒毛膜促性腺激素HCG)后激发其退化的性腺(卵巢和精巢)重新发育(与当年银鳗作对照).通过性腺组织切片观察产后鳗和对照鳗性腺发育成熟的全过程及其差异,发现产后鳗起初性腺发育比当年银鳗差,但经多次注射激素后,产后鳗性腺成熟与当年银鳗同步,证明产后鳗生殖细胞对激素的敏感件高.应用17α,20β-双羟孕酮和促性腺激素释放激素类似物(GnRH-A3)使催熟的产后鳗和对照鳗均产卵和排精,并孵化出仔鱼,从而有力地证明,鳗鲡产后虽体质弱,但待体能恢复后能够继续生存和繁殖.本研究旨在探讨利用产后鳗作为今后鳗鲡人工繁殖亲鳗的可行性.     

Effects of salinity and temperature on the growth and survival of New Zealand shortfin,Anguilla australis,and longfin,A. dieffenbachii,glass eels

The ideal water conditions for maximizing the performance of the nursery culture of glass eels harvested from the wild for aquaculture need to be determined for the New Zealand shortfin (Anguilla australis) and longfin (Anguilla dieffenbachii) eels. This study determined the survival and growth of glass eels reared under different temperature and salinity conditions in the laboratory. The growth and survival of shortfin and longfin glass eels reared in salt water (35‰) maintained at 25 °C was examined over 84 days from capture. The mean specific growth rate (SGR) was higher in shortfin [2.30±0.29% body weight (b.w.) day^?1] than longfin glass eels (1.52±0.06% b.w. day^?1), and survival was also higher in shortfin (76.0±4.16%) than for longfin glass eels (28.7±6.36%). A second experiment identified the effect of salinity (0, 17.5‰ and 35‰) and temperature (17.5 and 26.5 °C) on the acclimation, growth performance and survival of shortfin and longfin glass eels over a period of 84 days from capture. There was no incidence of mortality for either shortfin or longfin glass eels reared across all salinity treatments (0‰, 17.5‰ and 35‰) at 26.5 °C, while survival of shortfin and longfin glass eels reared at 17.5 °C was the highest in 17.5‰, followed by 35‰ and 0‰ treatments. Both temperature and salinity affected the SGR of shortfin glass eels, with the highest SGR observed for shortfin glass eels reared in 0‰ water maintained at 26.5 °C. In longfin glass eels, salinity alone had an effect on the SGR, with the highest SGR observed in glass eels reared in 0‰ water regardless of the water temperature (17.5 and 26.5 °C). In addition, the adaptability of glass eels to salinity was evaluated from the development and the physiological responses of gill chloride cell (CC) morphology. The number and size of CCs increased significantly with increasing salinity in both shortfin and longfin eels.     

American eel, Anguilla rostrata, growth in fresh and salt water: implications for conservation and aquaculture

Abstract  Growth of American eels, Anguilla rostrata (Lesueur), sampled in saltwater bays and adjacent freshwater ponds in the southern Gulf of St. Lawrence, Canada, was estimated by otolith annulus back-calculation, and related to salinity of growth habitat as determined using otolith strontium:calcium ratios. Eels resident in salt water grew on average in length 2.2 times faster than freshwater residents and increased in weight 5.3 times faster than freshwater residents. Previous findings of superior American eel growth rate in brackish water are thus extended to full-strength salt water. Freshwater residents took an estimated 2.4 times longer than saltwater residents to reach the silver eel stage. The extended time to maturation of eels reared in fresh water increases their vulnerability to fisheries and to bio-accumulation of contaminants. Freshwater eel populations should be managed more conservatively than those in saline waters. The increase in eel growth with salinity suggests that rearing in brackish or salt water could improve growth rates in eel aquaculture.     

Effects of steroid hormones on in vitro oocyte maturation in white sturgeon (Acipenser transmontanus)

The in vitro effects of several steroids on the maturation of intact white sturgeon (Acipenser transmontanus) ovarian follicles were investigated. At the highest concentration (1024 ng ml^–1 for the C21 steroids and 1139 ng ml^–1 for the C19 steroids), all of the C21 steroids tested, progesterone (P4), 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,(20,21-trihydroxy-4-pregnen-3-one 20-S), 11-deoxycortisol (S) and cortisol (F), as well as testosterone (T) induced germinal vesicle breakdown (GVBD) at 14 and 22 h. At 6 h, only P4 and 17,20-P induced maturation at the highest concentration (1024 ng ml^–1). At 14 and 22 h, 11-deoxycortisol was the most potent steroid inducer of GVBD followed by P4, 17OHP, 17,20-P, and 20-S. The steroid 11-hydroxytestosterone (11OHT) was completely ineffective at all concentrations and exposure times. The C21 steroids induced oocyte maturation at concentrations ranging from 4 to 1024 ng ml^–1, whereas T induced GVBD at 225 to 1139 ng ml^–1. Calculation of the mean effective concentration that induced 50% GVBD (EC50) from the 22 h incubations revealed the following order of potencies: S > P4 > 17OHP > 17,20-P > 20-S >> F > T. These bioassay results, together with previous findings on the endogenous production of steroids by ovarian follicles from gonadotropin-primed females, indicate that more than one steroid has a biological role in the resumption of meiosis in sturgeon oocytes and provides empirical evidence for P4, 17OHP, S, 20-S, and 17,20-P as maturation-inducing steroids in white sturgeon.     

Decreasing dietary lipids improves larval survival and growth of Japanese eel Anguilla japonica

Shark eggs-based diet is the only diet by which eel larvae can grow to glass eels in captivity. However, the high level of lipids in the diet is suggested to negatively affect eel larvae. This paper examines the effect of defatted shark eggs (DSE) and hen egg yolk (HY) on growth and survival of larvae of Japanese eel Anguilla japonica. Lyophilized shark egg and commercial HY were defatted with n-hexane, and four experimental diets were prepared using both defatted and untreated shark eggs and HY. Larvae were reared for 3 weeks by feeding the experimental diets. The highest survival rate was observed in the larvae fed DSE, and larvae fed HY showed the lowest survival rate. The best growth was found in larvae fed DSE, followed by shark eggs and defatted HY, and the worst growth was in HY-fed larvae. These results show that decreasing dietary lipids improves the nutritional value of both shark eggs and HY for eel larvae. Regulation of the dietary lipid level may positively affect the larval performance of eels by combination of ingredients with a low lipid content.