Sperm quality of Brazilian flounder Paralichthys orbignyanus throughout the reproductive season

The aim of this study was to evaluate the sperm quality of Brazilian flounder Paralichthys orbignyanus throughout its reproductive season. Sperm was collected at the beginning, middle and end of the breeding period. Spermatozoa density was maximum at the beginning (12.7 ± 0.92 × 10⁹ cells mL^?1) and at the end (11.8 ± 0.39 × 10⁹ cells mL^?1) of the breeding season (P<0.05). Sperm production and the percentage of spermatozoa moving fast forward increased significantly towards the end of the breeding season (P<0.05). The mean duration of progressive motility of spermatozoa was around 10 min. No difference was observed during the reproductive season in the percentage of motile cells, pH, osmolality and K⁺, Cl^? and Mg²⁺ concentrations in seminal plasma. The concentration of Na⁺ increased throughout the breeding season, reaching 174.62 ± 12.68 mmol L^?1 at the end (P<0.05). There was a decline in the concentration of Ca²⁺ (12.31 ± 3.08 mmol L^?1) in the middle of the breeding season, which coincided with the shortest motility duration of spermatozoa. The information reported in this study should help to improve management and optimize the development of protocols for short‐term storage and cryopreservation of Brazilian flounder semen.     

室内工厂化养殖犬齿牙鲆的生长特性

对室内工厂化养殖的犬齿牙鲆生长特性进行了研究。结果表明,其生长旺盛期在2龄,日生长最快达5.52g/d,第1年平均为1.65g/d,第2年平均为2.34g/d,第3年平均为1.3g/d,第4年平均为0.75g/d。养殖1周年其平均体重可达600g左右,两周年达1500g;其全长和体重的关系为W=0.0076TL3.1785(R2=0.9765),全长与体高和标准体长的关系分别为TL=2.5685BD-0.6594(R2=0.9823),TL=1.1957SL-0.1724(R2=0.9922)     

The potential impact of summer flounder (Paralichthys dentatus) aquaculture on regional flounder price

Abstract

Increasing consumer demand for flounder and dramatic reductions in ocean flounder harvests have raised interest in summer flounder (Paralichthys dentatus) aquaculture. Flounder farmers have several potential markets, including regional seafood dealers and high‐end niche (e.g., sashimi and ike‐jime) buyers. Prices paid by buyers are exogenous to typical aquacultural budgeting models, yet they are a key determinant of potential profitability. We develop a simple economic model to assess the impacts of aquaculture industry supply on prices paid by regional seafood dealers. The model is parameterized for a region with established flounder dealers and active field research in flounder production methods. We derive the price elasticity of regional seafood dealer demand for farmed flounder, and we examine the impacts of changes in production cost and ocean harvest on regional price and the level of aquacultural production that maximizes returns to the aquaculture industry.     

舟山褐牙鲆（Paralichthys olivaceus）仔鱼摄食和生长的研究

试验研究了褐牙鲆Paralichthys olivaceus仔鱼的摄食和生长。结果表明,仔鱼初次开口摄食时间在6日龄（16～18℃）,初次摄食率为30％,8日龄饥饿仔鱼最大初次摄食率为65％,孵出后第9天达到不可逆点（PNR）,混合营养期为3d左右。初次摄食强度和初次摄食率开始时较低,在卵黄囊耗尽时达最高,然后下降。随着饥饿时间的延长,仔鱼出现“负增长”,孵化第11天存活率仅为10％,12日龄全部死亡。     

Artificial spawning of carp, Cyprinus carpio (L.)

The effect of reproduction was investigated on females of Hungarian strain W, French strain F, and their cross‐breed 1X whose ovulation was stimulated with carp pituitary (0.3 mg kg^?1and, after 12 h, 2.7 mg kg^?1) or Ovopel (one‐fifth of a pellet per kg and, after 12 h, one pellet per kg). It was found that in the case of Ovopel treatment, the percentage of spawning females of strain F and the cross‐breed 1X was higher than in the hypophysed fish compared. The applied ovulation stimulators did not significantly affect the weight of obtained eggs, whereas the significant (P ≤ 0.01) effect was recorded with respect to the quality of eggs after 12‐, 24‐, 36‐ and 48‐h of incubation. After Ovopel stimulation, the quality of eggs was better. The origin of the females had no statistically significant effect on the weight of eggs although the yield of eggs from fish of strain W was much smaller than that from females of strain F and the 1X cross‐breed. The interaction between the ovulation stimulator and the provenance of the females was significant (P ≤ 0.05) for the percentage of live embryos after 48‐h of incubation of eggs. Eggs of the best quality (and highest weight) were obtained from fish of strain F and cross‐breed 1X treated with Ovopel. In females of strain F that spawned within 6 and 10 h after the second Ovopel injection, the effect of the ovulation time on the weight of eggs was non‐significant. It was significant with respect to the percentage of egg fertilization and of live embryos after 36‐h of incubation (P ≤ 0.01 and P ≤ 0.05 respectively). The better quality of eggs (and their higher weight) was recorded when this time was shorter.     

几种工业废水对牙鲆仔稚鱼的急性毒性及联合毒性作用

印染废水、电镀废水、农药废水都是常见的、易进入近岸海域的工业废水。本文通过室内实验，研究了这3种工业废水及其混合物对牙鲆仔稚鱼的急性毒性作用。结果表明，印染废水、电镀废水、农药废水及其混合物对牙鲆初孵仔鱼48hLC50(95％置信区间）和96h LC50（95％置信区间）分别为2．28％（1．39％－3．74％）、0．21％（0．18％－0．27％）、0．47％（0．37％－0．60％）及0．42％（0．07％－2．54％）和0．58％（0．15％－0．76％）、0．10％（0．04％－0．25％）、0．23％（0．18％－0．29％）及0．17％（0．12％－0．23％）；对牙鲆后期仔稚鱼48h LC50(95%置信区间）和96h LC50（95％置信区间）分别为2．95％（1．65％－5．29％）、0．22％（0．15％－0．31％）、0．71％（0．50％－1．01％）及0．77％（0．50％－1．19％）和0．69％（0．51％－0．93％）、0．14％（0．10％－0．20％）、0．38％（0．23％－0．61％及0．24％（0．16％－0．35％）。以对牙鲆仔稚鱼半致死浓度为指标，这几种工业废水的毒性顺序为电镀废水＞农药废水＞印染废水，初孵仔鱼较后期仔稚鱼对这些废水更为敏感。这些废水以等体积混合时，对初孵仔鱼和后期稚鱼的联合毒性作用表现为相加作用。     

牙鲆亲本对子代贡献率的实验研究

利用18对高度多态性微卫星标记, 对13尾牙鲆(Paralichthys olivaceus)亲鱼的188尾后代进行亲子鉴定, 计算其非亲排除率、鉴定准确率以及亲本对子代的贡献率。使用4个微卫星标记进行亲子鉴定, 排除率≥0.999 9, 准确率为92.02%; 用6个标记的排除率≥0.999 999, 准确率为96.81%; 用8个标记的排除率≥0.999 999 999, 准确率为97.87%; 用10个标记的准确率为99.47%; 用12～18个标记的准确率为100%。亲子鉴定准确率随非亲排除率和微卫星标记数目的增多而升高。检验所有子代全部是实验亲本的后代, 即13个亲本对子代均有贡献, 但不同亲本的贡献率不同, 最高为47.34%, 最低为0.53%。实验表明, 基于多标记的非亲排除率检测方法可以用于牙鲆亲子鉴定, 其鉴定准确率可以达到100%; 不同亲本对子代贡献率不同, 差异很大, 产生差异的原因有待进一步研究。

     

Genetic relationship between broodstocks of olive flounder, Paralichthys olivaceus (Temminck and Schlegel) using microsatellite markers

For the first generation of a selective breeding programme, it is important to minimize the possibility of inbreeding. This mostly occurs by mating between closely related individuals, while proper mating can provide an opportunity to establish the base families with wide genetic variation from which selection for subsequent generations can be more effective. Genotyping with microsatellite‐based DNA markers can help us determine the genetic distances between the base populations. The genetic markers further facilitate the identification of the correct parents of the offspring (parentage assignments) reared together with many other families after hatching. We established a genetic analysis system with microsatellite DNA markers and analysed the genetic distances of three farmed stocks and a group of fish collected from wild populations using eight microsatellite markers. The averaged heterozygosity of the farming stocks was 0.826 and that of the wild population was 0.868. The hatchery strains had an average of 8.6 alleles per marker, which was less than a wild population that carried an average of 14.3 alleles per marker. Significant Hardy–Weinberg disequilibrium (HWDE) was observed in two farming stocks (P<0.05). Despite relatively low inbreeding coefficiency of the hatchery populations, the frequency of a few alleles was highly represented over others. It suggests that the hatchery stocks to some extent have experienced inbreeding or they originated from closely related individuals. We will develop a selective program using the DNA markers and will widen the usage of the DNA‐based genetic analysis system to other fish species.     

克隆牙鲆和普通牙鲆血液生理生化指标的比较

以中国水产科学研究院北戴河中心实验站养殖的双单倍体(DH)牙鲆(Paralichthys olivaceus)作为亲鱼, 采用冷休克方法抑制第二极体排出, 建立牙鲆克隆家系。以普通牙鲆的一个家系作为对照组, 在同样饲养条件下进行培育。实验组和对照组均为2⁺ 龄鱼, 克隆家系随机抽取5尾, 体质量(1 398±88) g; 普通群体随机抽取10尾, 体质量(1 749±125) g。分别采集血液进行血液生理和生化指标的比较分析。结果表明: 1) 克隆牙鲆的红细胞数量(RBC)、血红蛋白数量(HGB)、平均血红蛋白量(MCH)、红细胞压积(HCT)极显著低于普通牙鲆(P<0.01), 而在其他血液生理指标上二者没有显著性差异(P>0.05); 2) 克隆牙鲆在白球比(ALB/GLB)、谷草转氨酶/谷丙转氨酶(AST/ALT)比值、谷草转氨酶(AST) 3个指标上显著高于普通牙鲆(P<0.05), 其中AST差异达到极显著水平(P<0.01), 而碱性磷酸酶(ALP)、总胆固醇(CHO)、尿素氮(BUN) 3项指标, 克隆牙鲆极显著低于普通牙鲆(P<0.01)。克隆牙鲆血液生理指标的平均变异系数为5.50%, 普通牙鲆的则为9.21%, 其中克隆牙鲆红细胞平均体积(MCV)的变异系数仅为1.52%。克隆牙鲆血液生化指标的变异系数平均为10.56%, 普通牙鲆的则为27.23%, 达到克隆牙鲆的2.58倍。结果证实, 克隆牙鲆个体间的一致性较好, 作为指标生物, 效果优于普通牙鲆等海产鱼类。

     

Effects of starvation on kidney melano-macrophage centre in olive flounder, Paralichthys olivaceus (Temminck and Schlegel)

We investigated the influence of nutritional conditions on histological changes in melano‐macrophage (MM) accumulation in the kidney caused by long‐term starvation. These data were used to determine nutritional indices for olive flounder, Paralichthys olivaceus (Temminck and Schlegel). Experimental groups were established: control, fed and starved. All fish were fed daily with commercial feed at 1–3% of their total body weight for 2 weeks before the start of the experiment (initial control). The control group received feed at 1–3% of their total body weight and the fed group received feed ad libitum, while the starved group fasted throughout the experiment. Ten fish were removed from the initial control at the start of the experiment and each of the three groups every 4 weeks during the 12 weeks of the experiment to examine to investigate kidney MM accumulation. Kidney MMs with dark brown pigment were randomly distributed in the kidneys of starved fish, increasing rapidly after week 4, while deposition levels remained low throughout the experiment in the control and fed groups. These results suggest that catabolic tissue breakdown is a major factor contributing to the formation of pigments within MMs. These results suggest that the degree of MMs deposition in the kidney can be used as alternative indicators to identify starvation in wild and cultured olive flounder.     

Pathogenicity of Edwardsiella tarda to olive flounder, Paralichthys olivaceus (Temminck & Schlegel)

The LD50 and cytotoxic and enzymatic activities of both cells and extracellular products (ECPs) of eight Edwardsiella tarda strains were determined and their bacterial superoxide dismutase gene (sodB) and catalase gene (katB) were sequenced. Strains were also examined for their ability to resist the immune responses of olive flounder, Paralichthys olivaceus. LD50 values of strains (FSW910410, KE1, 2, 3, 4, 5 and 6) in olive flounder ranged between 10(2.5) and 10(5.3) cfu (colony forming units) per fish. Unlike the avirulent strain SU100 (LD50>or=10(7)), all pathogenic strains were able to survive in flounder serum and head kidney leucocytes (except for KE2). The virulent strains possessed type I sodB and katB, whereas SU100 had type II sodB but not katB. However, there was no difference between avirulent and virulent strains in haemolytic and cytotoxic activities. The results of this study demonstrated that the ability of E. tarda to resist complement activity and phagocytosis is conferred by its superoxide dismutase and catalase, which thus play an essential role in the pathogenicity of this bacterium. In addition genotyping of sodB and kat B proved to be a very useful tool to distinguish virulent from avirulent strains.     

Distribution of three GnRHs in the brain and pituitary of the wild Japanese flounder Paralichthys olivaceus

ABSTRACT:   Wild adult maturing and immature female Japanese flounder Paralichthys olivaceus were collected in June 2004 and January 2005, respectively, to clarify a possible role of gonadotropin-releasing hormones (GnRHs) in reproduction. Levels of salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and sea bream GnRH (sbGnRH) in the brain and pituitary were examined by time-resolved fluoroimmunoassay. Three forms of GnRHs were detected in the discrete brain at various levels. In the pituitary of both maturing and immature fish, sbGnRH was abundant together with a pronounced amount of sGnRH, whereas cGnRH-II was almost below the detectable limit. In maturing fish, levels of sbGnRH were high in the telencephalon, hypothalamus and pituitary, while levels of sbGnRH of immature fish were very low in these regions. These results indicate that sbGnRH is mainly responsible for gonadotropin secretion, and that sbGnRH in the anterior part of the brain is associated with gonadal maturation in the Japanese flounder.     

Effects of high dose of vitamin A on reproduction and egg quality of Japanese flounder Paralichthys olivaceus

ABSTRACT: The present study was conducted to investigate the effects of dietary vitamin A on reproduction and egg quality in Japanese flounder Paralichthys olivaceus . Broodstock were fed experimental pellets containing two levels of vitamin A [11 × 10³ IU/100 g (control diet; CD), 337 × 10³ IU/100 g (experimental diet; ED)] for approximately 2 months before spawning and during the spawning period. Two groups of five females (average weight 1.4 kg) and 10 males (average weight 0.7 kg) were randomly allocated to two 30 m³ indoor tanks. Total egg production of the CD group was slightly higher than the ED group. Percentage of buoyant eggs and hatching rate of the ED group were significantly higher than the CD. In other egg quality parameters, such as percentage abnormal larvae and starvation tolerance of larvae, no notable difference was found between these two groups. At the end of the experiment, the skin color of broodstock in the ED group was darker than that of the CD group. Vitamin A content in eggs of the ED group was significantly higher than that of the CD group. However, the difference in vitamin A content in eggs between the ED and CD groups was much smaller than that in the liver of the females between the two groups. These results indicate that feeding broodstock a higher level of vitamin A increases the vitamin A content in eggs but does not affect egg quality in Japanese flounder because excess dietary vitamin A was stored mainly in the broodstocks' liver.     

Artificial spawning of carp (Cyprinus carpio L.): the use of Aquaspawn and carp pituitary to induce ovulation in females of Lithuanian line B

Ovulation stimulation was carried out in carp (Cyprinus carpio L.) females of Lithuanian line B, using carp pituitary (0.3 + 2.7 mg kg^?1), Aquaspawn (0.5 mL kg^?1) and Aquaspawn + carp pituitary (0.3 mL kg^?1 + 2.7 mg kg^?1). The poorest results of reproduction (the lowest percentage of ovulating females, the lowest weight of eggs and their poorest quality) were obtained after double hypophysation. The best results were induced by the application of Aquaspawn alone. The effect of the applied stimulators was statistically non‐significant with respect to the weight of eggs, being highly significant (P ≤ 0.01) with respect to the percentage of fertilized eggs and that of living embryos after 24‐ and 36‐h incubation. The multiple‐regression equations were calculated for the three groups separately. The percentage of living embryos after 36‐h incubation constituted a dependent variable while the mass of females, the mass of eggs, the fertilization percentage and the percentage of living embryos after 24‐h incubation were independent variables. The regression was only significant (P ≤ 0.01) for the group of fish treated with Aquaspawn.     

牙鲆头肾巨噬细胞的分离培养与鉴定

本研究使用密度梯度离心法从牙鲆(Paralichthys olivaceus)头肾中分离得到了巨噬细胞,通过差速贴壁法对获得的细胞进行纯化,后续经过优化培养条件和培养过程,采用L-15培养基(Gibco)、5％胎牛血清、1％青–链霉素、1％非必需氨基酸、30％L929细胞培养基在24℃、无CO2的条件下进行培养.显微镜...     

Induction of ovulation in ornamental common carp (Koi, Cyprinus carpio L.) using LHRHa ([d-Ser(tBu)6, Pro9-NEt]-LHRH) combined with haloperidol and carp pituitary extract

The effects of a single injection of mammalian superactive analogue [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa (20 μg/ kg^?1) combined with the dopamine antagonist, haloperidol (HAL, 0.5 mg kg^?1), for induction of ovulation in the koi carp broodstocks were determined under routine hatchery conditions. The results were compared with classic carp pituitary extract (CPE, double injection) application (water temperature 22 °C). Physiological saline (0.7% NaCl)‐injected fish were used as a control group and no ovulation occurred in this group. The spawning ratio was high in the LHRHa+HAL treatment group and in the CPE treatment group (6/7 and 5/7 respectively). The latency period was 14–16 h in the LHRHa+HAL treatment group and 12–14 h in the CPE treatment group (after the second injection). There was no difference between the two ovulating groups with respect to the spawning index (the weight of eggs as a percentage of female body weight) and fertilization rate of eggs (P>0.05). As a result, ovulation can be induced successfully in koi carp broodstocks with 20 μg kg^?1 [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa+0.5 mg kg^?1 HAL treatment in a single injection without decreasing the egg quality. Application of this combination can be useful for hatchery and broodstock management in koi carp culture.     

牙鲆体质量与形态性状的异速生长分析

为分析牙鲆（Paralichthys olivaceus）体质量与形态性状之间异速生长的遗传规律,通过人工受精的方法建立牙鲆19个全同胞家系,测量了牙鲆不同日龄的体质量和形态性状。采用逐步回归方法建立最优联合异速生长模型,将此模型镶嵌到体质量动物模型的每个遗传和环境效应中,进一步分析对多个形态性状异速生长遗传规律。静态异速生长分析表明：体质量与全长之间存在最大异速生长指数（1.415 5）,表现为正异速生长,剩余形态性状与体质量之间的异速生长指数为0.061 5~0.718 0,皆表现为负异速生长。全长与尾柄高之间异速生长指数的正遗传相关最大（0.907 8）;全长与尾柄长之间的负遗传相关最大（0.946 8）。不同模型通过统计标准比较,确定模型Ⅰ为进行牙鲆动态异速生长分析的最优随机回归模型。     

Dietary myo‐inositol requirement for Olive flounder,Paralichthys olivaceus (Temminch et Schlegel)

This study was aimed at examining the essentiality and requirement of inositol for Olive flounder, Paralichthys olivaceus. Six casein–gelatin‐based semi‐purified diets were formulated to contain five different levels of myo‐inositol (MI) (designated as M0, M0+, M200, M400, M800 and M1600 for 0, 0+antibiotic, 200, 400, 800 and 1600 mg kg^?1 respectively). After 20 weeks of a long‐term feeding trial, fish (initial body weight, 10 g) fed the M800 diet showed significantly increased growth performances and survival. Liver lipid concentration tended to decrease as the dietary MI increased, although there was no significant difference among all treatments. Polyunsaturated fatty acid in the liver of the fish fed the diets containing high levels (M1600) of MI was significantly increased. Hepatic inositol concentration of the fish was significantly increased by dietary MI supplementation. The intestinal biosynthesis of inositol by microflora seemed to be insufficient to prevent growth retardation in juvenile Olive flounder. The optimum level of dietary MI for juvenile Olive flounder was found to be 617 mg kg^?1 based on weight gain in a broken‐line regression model. This finding indicates that MI supplementation is required at a concentration of at least 617 mg kg^?1 to maximize the growth performances and to prevent abnormal lipid metabolism.     

Dietary choline requirement of juvenile olive flounder (Paralichthys olivaceus)

An eight‐week study was conducted to determine the optimum dietary choline level in juvenile olive flounder, Paralichthys olivaceus. Seven diets were prepared to contain 0, 250, 500, 750, 1,000, 2,000 and 3,000 mg/kg diet. Juveniles (5.9 ± 0.03 g; 5.5 ± 0.4 cm; mean ± SD) were randomly distributed into 21 tanks (25 fish/tank) and fed one of the diets in triplicates. Survival rate of fish fed the diet containing the lowest choline level was significantly lower than those of fish fed the other diets (p < 0.05). Final body weight, weight gain, specific growth rate, feed efficiency and protein efficiency ratio significantly increased with increasing choline levels up to 1,000 mg/kg diet. Whole‐body protein and lipid contents increased in accordance with choline levels up to 750 mg/kg diet, beyond which they plateaued. Liver and muscle lipid contents elevated with increasing choline levels up to 2,000 mg/kg diet. Plasma cholesterol esters, triglycerides, cholesterol and total lipids were significantly influenced by the graded choline levels; however, responses of those indices were not identical. Broken‐line analyses of weight gain and liver choline concentrations responding to the graded choline levels revealed that choline requirements of the juvenile flounder could be between 847 and 1,047 mg/kg diet.