Fast development of genetically uniform strains by gynogenesis in Japanese flounder (Paralichthys olivaceus)

The Japanese flounder is a major species in both aquaculture and research. Inbred strains of Japanese flounder were developed efficiently in our laboratory by meiotic and mitotic gynogenetic reproduction techniques. To determine the induction efficiency of gynogenesis and the rate at which full homozygosity is produced, six meiotic gynogenetic females (G1–G6) that had experienced meiotic gynogenesis once and three common females (C1–C3) were selected for production of mitotic gynogenesis in our experiments. Of the nine adult females, all six gynogenetic fish successfully produced viable offspring. However, only one of the three common fish did. Using microsatellite markers, we estimated the homozygosity of gynogenetic fish induced by mitotic gynogenesis. We found that the homozygosity of seven lines (C1, G1–G6) increased quickly, individually from 43%, 72%, 69%, 71%, 78%, 69% and 58% to 100%, 100%, 100%, 90%, 100%, 100% and 90% offspring complete homozygosity. Under mitotic gynogenesis, individuals with higher homozygosity had a higher induction rate. The G4 line showed the highest induction rate, achieving 44.59% convert hatching rate and 29.28% convert normality rate. We conclude that meiotic gynogenesis may be a feasible method to produce DH genetic material in Japanese flounder.     

杂交鲆(牙鲆♀×夏鲆♂)网箱养殖技术研究

引进杂交鲆苗种在宁德市三都海区,经过385d的网箱养殖,达到商品规格,平均全长36.3cm,平均体重606g,成活率达71%,饵料系数为8.3.研究表明,杂交鲆在生长速度、抗逆性、适应性等方面有较明显的优越性,能适应闽东海域夏季高温和冬季持续的低温.以Keys氏公式W=aLb,拟合养殖13个月杂交鲆的平均全长L(cm)...     

甲氧苄氨嘧啶在牙鲆体内的药代动力学研究

健康牙鲆口服剂量为100 mg/kg的甲氧苄氨嘧啶后,利用高效液相色谱测定药物在牙鲆肌肉、血液、肾脏、肝脏质量浓度变化,并通过3P87药动学软件分析数据。试验结果表明,甲氧苄氨嘧啶在肌肉、血液、肾脏、肝脏4种组织中均符合一级吸收二室开放模型,其在上述4种组织中的达峰时间(tmax)分别为18.72、12.942、3.13 h和19.49 h;达峰质量浓度分别为13.80、18.23、32.79、27.40μg/ml;消除半衰期分别为23.64、21.471、6.72 h和15.99 h。口服甲氧苄氨嘧啶,在牙鲆体内吸收与消除较慢,血药浓度高,维持时间长。     

Interactive effects of dietary cholesterol and protein sources on growth performance and cholesterol metabolism of Japanese flounder (Paralichthys olivaceus)

A feeding trial was conducted to compare the effects of supplemental cholesterol in fish meal (FM), fish protein concentrate (FPC), soy protein isolate (SPI) and soy protein concentrate (SPC)‐based diets on growth performance and plasma lipoprotein levels of Japanese flounder (Paralichthys olivaceus). Eight isonitrogenous and isocaloric diets including FM, FPC, SPI or SPC as sole protein source with or without supplementation with 10 g cholesterol kg^?1 diet were fed to juvenile fish for 8 weeks. Dietary cholesterol supplementation significantly increased the feed intake and specific growth rate in fish fed SPI‐based diets, but decreased those in fish fed FPC‐based diets. In addition, cholesterol supplementation significantly increased the level of cholesterol and ratio of low‐density lipoprotein cholesterol to high‐density lipoprotein cholesterol in plasma of fish fed fish protein‐based diets, whereas no effects were observed in fish fed soy protein‐based diets. The hepatic lipid content of fish fed FPC‐, SPI‐ or SPC‐based diets were significantly increased by supplemental cholesterol, but no influence was observed in fish fed FM‐based diets. These results suggested that dietary protein source modify the growth‐stimulating action of cholesterol; cholesterol supplementation may increase the arteriosclerotic lesion in fish fed fish protein‐based diets and the incidence of fatty liver in fish fed soy protein‐based diets.     

牙鲆NR4A1基因在甲状腺素诱导仔鱼变态中的表达调控分析

NR4A1基因是一个重要的转录因子,在信号转导早期通过对靶基因的特异性转录调控发挥着重要的作用。从牙鲆(Paralichthys olivaceus)中克隆了NR4A1基因c DNA序列,检测了其在牙鲆变态中和成鱼各组织中的表达模式,分析了其在外源甲状腺素(TH)以及硫脲(TU)处理仔鱼中的表达变化。结果表明,牙鲆NR4A1 c DNA全长3264 bp,编码568个氨基酸;牙鲆NR4A1基因与其它物种具有很高的同源性,在系统进化树中与鱼类聚为一支;牙鲆NR4A1基因在成鱼心、肌肉和鰓中高表达;在变态过程中,NR4A1水平逐渐升高,且在25 dph达到最高,之后逐渐降低;TH组中的NR4A1基因水平在变态早期和高峰期显著低于正常组,TU组的水平在变态中后期和结束期显著高于正常组;TU组中变态被抑制的仔鱼在正常海水和0.1 mg·L~(-1)TH海水中饲养6 d后,能够顺利变态,且NR4A1基因在拯救组(TU抑制变态仔鱼在正常海水和0.1 mg·L~(-1)TH海水中饲养)中的表达水平与TU对照组相比显著降低,与正常组中同时期的水平一致。结果表明,牙鲆NR4A1基因可能在仔鱼变态调控中扮演着非常重要的角色。     

不同蛋白水平对牙鲆能量收支的影响

鱼类的能源物质以蛋白质和脂肪为主,但在鱼体内蛋白质代谢的最终产物以氮排泄物形式通过鳃排出体外,少量随尿排出,氮排泄的主要成分为氨和尿素,仅有少量用于生长.本实验采用鱼类生物能量学的方法,在保持最大摄食率水平的条件下,通过四个不同梯度蛋白质对牙鲆个体的能量转换进行了研究,从而探讨牙鲆的最适蛋白水平,结果表明:当牙鲆投喂蛋...     

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.     

牙鲆变态中两种HSP90基因的不同表达及其与甲状腺激素的关系

热休克蛋白90(heat shock protein90,HSP90)是细胞内主要的伴侣蛋白,在激素信号转导、细胞分化、细胞增殖与凋亡、形态发生与进化、变态发育及应激防御等多重调节路径中发挥关键作用。采用实时荧光定量RT-PCR技术检测了牙鲆两种HSP90基因在仔鱼发育和在成年组织中的表达变化。结果表明,HSP90α在成鱼骨骼肌、肠和胃中有较高的表达,而HSP90β在成鱼脑、脾脏和肾脏中有较高的表达。HSP90αmRNA水平在仔鱼变态期间迅速增加,至变态高峰G期达到最高水平;相反,HSP90β转录在仔鱼整个变态期间变化不是特别明显。鉴于甲状腺激素(thyroid hormone,TH)在牙鲆变态中的重要作用,还运用外源的TH及硫脲(thiourea,TU)处理牙鲆仔鱼来确定TH对HSP90基因转录的调节。与未处理组相比,HSP90α转录在TH处理8d和13d的仔鱼中显著增加,而在TU处理仔鱼中明显减少;但HSP90βmRNA水平不受药物处理影响。从上述结果分析,牙鲆中HSP90α的转录很可能被甲状腺激素上调,而且其在牙鲆的变态发育中发挥了重要作用。     

Effect of stocking density and feeding regime on larval growth,survival, and larval development of Japanese flounder,Paralichthys olivaceus,using live feeds

This research examined the effect of initial stocking density and feeding regime on larval growth and survival of Japanese flounder, Paralichthys olivaceus. Larval rearing trials were conducted in nine 50‐L tanks with different initial stocking densities combined with different feed rations (20 larvae/L with standard feed ration [LD], 80 larvae/L with standard feed ration [HD], and 80 larvae/L with four times the standard feed ration [HD+]). Larvae were stocked on 0 days posthatch (DPH) following hatching of the fertilized embryos. Larval total length (TL), survival rates, and final densities were observed on larval settlement (32 DPH) to evaluate larval rearing performance. At 32 DPH, there were no significant differences (p > .05) in TL or survival rates between the LD (46.5 ± 17.0%) and HD+ (40.3 ± 9.4%). The TL and survival rate of HD (23.1 ± 3.5%) were significantly lower than that of LD and HD+ (p < .05). However, the larval density of HD was significantly higher than that of LD (p < .05). HD+ achieved the best larvae production (32.27 ± 7.51 larvae/L), supported by sufficient food source, high water exchange, and proper water quality management (routine siphoning, surface skimming). The larval‐rearing protocols and larval development from hatching to metamorphosis is described in detail, with corresponding photographs taken during the experiment.     

牙鲆群体生化遗传学研究— — Ⅱ．等位酶的生化遗传分析

应用聚丙烯酰胺凝胶电泳技术对牙鲆[Paralichthys olivaceus(T.et S.)]养殖群体进行10种等位酶的电泳检测和谱带遗传分析,确定了21个等位酶位点和25个等位基因,单态位点有Est-2、Est-3、Aat-2、Sod-1、Sod-2、Ldh-1、Mdh-1、Me-1、Adh-2、Sdh-2、Sdh-3、Idh-1、Idh-2、Amy-1、Amy-2和Amy-3,而这些位点仅有1个等位基因。仅有Aat-l、Sdh-1、Adh-l、Est-1和Est-4(P_(0.99)标准)5个位点是多态的,多态位点的百分数为23.81％,而这些位点有2个等位基因。揭示了牙鲆群体等位酶位点及其等位基因带谱的变异式样,为牙鲆遗传育种及遗传结构的研究提供一批等位酶位点及其等位基因参考图谱。