异源精子诱导犬齿牙鲆的雌核发育

用冷冻保存的花鲈精子作为异源精子,采用紫外线(UV)对精子进行照射使其遗传物质失活,然后与卵子进行"授精",可以刺激犬齿牙鲆鱼卵进行雌核发育,在受精后一定时间采用冷休克处理"授精"卵,抑制第二极体排出成功获得了犬齿牙鲆雌核发育二倍体鱼苗.实验表明:犬齿牙鲆同源精子经80 mJ/cm2的紫外线照射可以完全失活,冻存花鲈精子经紫外线照射后也同样具有诱导犬齿牙鲆雌核发育的能力.无论同源精子还是异源精子,最佳诱导条件为在18℃条件下,精子经80 mJ/cm2的紫外线照射,受精后4～5 min将受精卵放在3℃海水中进行冷休克处理,持续时间为45 min,同源精子和异源精子二倍体诱导分别为32.66%±7.03%和28.00%±6.48%.采用形态学、流式细胞仪DNA含量分析和微卫星标记技术对雌核发育鱼苗进行了分析,证明了雌核发育鱼苗为雌核发育二倍体.     

4种海水鱼淋巴囊肿组织的病理特征比较

取山东、河北、浙江等地感染淋巴囊肿病的牙鲆(Paralichthys olivaceus)、许氏平鮋(Sebastes schlegeli)、鲈鱼(Lateo-labrax japonicus)及纹腹叉鼻(Arothron hispidus),利用光镜和电镜技术及组织化学方法,对患病鱼淋巴囊肿组织的病理特征进行观察比较。结果发现,来自不同地区同一种鱼的淋巴囊肿组织的病理特征无明显差异,不同种鱼的淋巴囊肿细胞具有共同的特征:细胞膨大,细胞核不规则,细胞质内有嗜碱性的、呈Feulgen和Mann氏反应阳性的包涵体,囊肿细胞的细胞膜外有呈PAS反应阳性的均质囊壁,细胞质内病毒颗粒的大小200～220 nm,核周池内有高电子密度物质等。不同种鱼囊肿组织细胞的大小、细胞核的不规则程度、细胞质内包涵体的形态、细胞质内病毒粒子的分布状态,以及囊肿物的外观等有差异。虽然不同种鱼之间存在差异,囊肿组织共同的病理学特征仍可作为疾病诊断的可靠依据。     

环境胁迫对许氏平鲉和花鲈血糖、血沉降的影响

研究了运输、温度和空气暴露胁迫对许氏平鲉(Sebastes schlegeli)和花鲈(Lateolabrax maculatus)血液生化指标的影响。60min长途运输使许氏平鲉和花鲈的血糖含量都有所上升，其中许氏平鲉的血糖含量在运输后24h基本恢复到正常水平，而花鲈的血糖水平在168h后可恢复到稳定状态。季节温度变化是引起春冬两季许氏平鲉血糖水平在经运输胁迫后产生差异的主要原因，春季(4月)经运输胁迫后鱼体血糖含量约为冬季(11月)的1．4倍。在温度梯度实验中，设定不同温度组(10℃、15℃、20℃、23．5℃、26℃)检测许氏平鲉血糖和血沉降的变化，血糖值出现峰值，而血沉降则随温度升高一直表现出上升趋势。在温度梯度实验基础上进行的空气暴露实验中，各胁迫组的血糖含量升高，血沉降轻微降低但整体变化趋势与温度梯度实验结果相似。     

鲈鱼(Lateolabrax japonicus)生长后期对苏氨酸需要量

在半精制饲料中分别添加0、0.35%、0.70%、1.05%、1.40%、1.75%苏氨酸,制成苏氨酸实际梯度为1.05%、1.35%、1.65%、2.00%、2.42%、2.65%的6组等能等氮饲料(44.67%粗蛋白质,21.65 k J/g总能),对初始体重为(333.93±6.60)g的鲈鱼(Lateolabrax japonicus)在海水浮式网箱(1.5 m×1.5 m×2.0 m)中进行了70 d的喂养实验,研究其对苏氨酸的最适需求量。结果显示,鲈鱼成活率在89.58%–95.83%之间,各处理组之间无显著差异(P0.05);随着饲料中苏氨酸水平的升高,鲈鱼的特定生长率(SGR)显著增加(P0.05),且在2.00%苏氨酸饲料组出现最大值,但随着苏氨酸水平的继续升高,SGR呈减小的趋势;饲料效率(FE)随饲料中苏氨酸水平的升高呈先增加后减小的趋势,2.00%苏氨酸组的FE显著高于1.05%组及2.65%组(P0.05);随着饲料中苏氨酸水平的升高,蛋白质沉积率(PPV)呈先增加后减小的趋势,且于2.00%苏氨酸组出现最大值;肝脏谷草转氨酶、谷丙转氨酶活性随饲料中苏氨酸水平的升高呈先增加后减小的趋势;饲料中不同水平苏氨酸对鱼体粗蛋白、粗脂肪、粗灰分无显著影响(P0.05)。以特定生长率、饲料效率及蛋白质沉积率为评价指标,经二次回归分析得出,鲈鱼对饲料中苏氨酸的最适需求量分别为占饲料干重的1.84%、1.87%及1.83%,占饲料蛋白质的4.11%、4.18%及4.09%。     

Pharmacokinetics of oxolinic acid and oxytetracycline in kuruma shrimp, Penaeus japonicus

The pharmacokinetics of oxolinic acid and oxytetracycline were examined in kuruma shrimp (Penaeus japonicus) after intra-sinus (10 and 25 mg/kg, respectively) and oral (50 mg/kg) administration. The shrimp were kept in tanks with recirculated artificial seawater at a salinity of 22–23 ppt. The water temperature was maintained at 25±0.6 °C. The hemolymph concentrations of both drugs after intra-sinus dosing were best described by a two-compartment open model. The distribution and elimination half-lives (t_1/2 and t_1/2β) were found to be 0.59 and 33.2 h for oxolinic acid and 0.45 and 24.7 h for oxytetracycline, respectively. The apparent volume of distribution at a steady state (V_ss) and total body clearance (CL_b) were estimated to be 1309 ml/kg and 28.8 ml/kg/h for oxolinic acid and 748 ml/kg and 22.7 ml/kg/h, respectively. The hemolymph concentration–time curves after oral administration did not fit by the nonlinear least squares method using one- and two-compartment model with first-order absorption in either of the drugs. The peak hemolymph concentration (C_max), the time to peak hemolymph concentration (t_max) and the elimination half-life were found to be 17.8 μg/ml, 7 h and 34.3 h for oxolinic acid and 24.3 μg/ml, 10 h and 33.6 h for oxytetracycline, respectively. The bioavailability (F) after oral administration was 32.9% for oxolinic acid and 43.2% for oxytetracycline. The hemolymph protein binding in vivo was determined to be 36.7±8.5% for oxolinic acid and 22.9±4.8% for oxytetracycline.     

大连地区仿刺参养殖池塘叶绿素a分布和初级生产力估算

于2007、2008年每年的3-11月,对大连沿海地区的仿刺参养殖池塘叶绿素a含量进行了调查,并对初级生产力进行了估算.结果表明,大连沿海各地区的仿刺参养殖池塘中叶绿素a年均含量为2.11～5.40 mg/m3,最高值出现在2007年8月的庄河,平均值为12.96 mg/m3,最低值出现在2007年3月的旅顺附近,平均值为0.17 mg/m3.仿刺参养殖池塘中初级生产力的年均值为111.38～272.58 mg/(m2 · d),最高值出现在2007年6月的瓦房店复州湾地区,平均值为602.72 mg/(m2 · d),最低值出现在2007年3月的旅顺附近,平均值为16.47 mg/(m2 · d).     

日本对虾抗白斑病子三代的抗白斑综合征病毒感染能力及免疫特性

为了更好地进行日本对虾抗病良种的选育,本文用浓度为每毫升1×105个病毒粒子的对虾白斑综合征病毒(white spot syndrome virus,WSSV)粗提液注射感染抗白斑病日本对虾子三代和普通日本对虾,观察其死亡率,同时测定WSSV感染后0,4,24,48和96 h血淋巴酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)活力和蛋白含量.结果显示:注射WSSV 14 d内抗病子三代和普通对虾的平均死亡率分别为15%和85%,前者的抗病保护率达82.4%;感染前前者的各项免疫指标均高于后者,且PO,AKP和蛋白含量存在极显著差异(P＜0.01);感染后两者血淋巴检测的5项指标在96 h内的变化趋势基本相似,除ACP以外,抗病子三代的其他指标都比普通日本对虾的高.研究结果表明,所选育的抗白斑病日本对虾子三代能够有效地抵抗对虾白斑综合征病毒的侵染,其免疫指标也高于普通日本对虾.     

Survival and serum biochemical responses of spotted sea bass Lateolabrax maculatus during simulated waterless live transportation

Waterless live fish transportation is an alternative and promising transport strategy, which may reduce shipping costs, increase stocking density and improve survival of some species. Spotted sea bass is one of the most economically important marine fish in China. However, waterless transportation has not yet been studied on spotted sea bass. To explore the possibility of waterless transportation of spotted sea bass and its sublethal stress responses, the survival rate and dynamics of blood biochemical parameters were evaluated in the present study by preserving it under no water, less water and water (control) conditions for 9 hr. The serum biochemical parameters including total protein, cortisol, glucose, lactate, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total carbon dioxide (TCO₂), creatinine, blood urea nitrogen (BUN), ammonia, superoxide dismutase (SOD) activity and malonaldehyde (MDA) were investigated. Results showed that the survival rates of fish in all groups gradually declined with time increasing. At the end of transportation, the survival rates in the control group, less‐water group and no‐water group decreased to 49.11%, 15.60% and 23.96% respectively. Significant changes were observed in several selected serum biochemical parameters, including glucose, lactate, ALT, TCO₂, BUN and ammonia during simulated waterless transportation, which might negatively affect the physiological homeostasis of fish, leading to the high mortality in present study. In general, waterless transportation of spotted sea bass was possible, but it might be rather suitable for short time than long time.     

应用间接荧光抗体技术快速检测花鲈病原菌——鳗弧菌

以花鲈（Lateolabrax japonicus)弧菌病的病原菌-鳗弧菌（Vibrio anguillarum)W-1为抗原，制备兔抗血清；利用异硫氰酸荧光素标记的羊抗兔免疫球蛋白（FITC-IgG）为荧光标记二抗，并以罗丹明标记的牛血清白蛋白为背景染色，建立检测鳗弧菌的间接荧光抗体快速检测技术。应用该技术对人工感染后的花鲈组织（肌肉、鳃、肠、肾）样品和养殖水体样品进行了鳗弧菌检测，结果显示间接荧光抗体技术不仅可以用于诊断发病的感染花鲈，也可用于检测带菌状态或未发病的感染花鲈。     

Effects of body size and water temperature on food consumption and growth in the sea cucumber Apostichopus japonicus (Selenka) with special reference to aestivation

To investigate the effects of body size and water temperature on feeding and growth in the sea cucumber Apostichopus japonicus (Selenka), the maximum rate of food consumption in terms of energy (C_maxe; J day^?1) and the specific growth rate in terms of energy (SGRe; % day^?1) in animals of three body sizes (mean±SE) – large (134.0±3.5 g), medium (73.6±2.2 g) and small (36.5±1.2 g) – were determined at water temperatures of 10, 15, 20, 25 and 30°C. Maximum rate of food consumption in terms of energy increased and SGRe decreased with increasing body weight at 10, 15 and 20°C. This trend, however, was not apparent at 25 and 30°C, which could be influenced by aestivation. High water temperatures (above 20°C) were disadvantageous to feeding and growth of this animal; SGRe of A. japonicus during aestivation was negative. The optimum temperatures for food consumption and for growth were similar and were between 14 and 15°C, and body size seemed to have a slight effect on the optimal temperature for food consumption or growth. Because aestivation of A. japonicus was temperature dependent, the present paper also documented the threshold temperatures to aestivation as indicated by feeding cessation. Deduced from daily food consumption of individuals, the threshold temperature to aestivation for large and medium animals (73.3–139.3 g) was 24.5?25.5°C, while that for small animals (28.9–40.7 g) was between 25.5 and 30.5°C. These values are higher than previous reports; differences in sign of aestivation, experimental condition and dwelling district of test animals could be the reasons.