注射含HCG和CPE的W／O／W复乳诱导日本鳗鲡性腺发育成熟的研究

用含HCG及鲤脑垂体（PG）匀浆液提取物（CPE）的W／O／W复乳，对塘养雌鳗进行注射催熟实验，雌鳗平均GSI达35．18％，最高达56．5％，其催熟效果较常规注射方法（平均GSI 27．9％，最高50．4％）好，用含LHRH－A，HCG，CPE的复乳和DOM生理盐水混悬剂对部分达到性成熟的鳗鲡进行催产，用W／O／W复乳诱导成熟的鳗鲡，排卵率为75％，用常规方法催熟的鳗鲡排卵率为55％，注射复乳制剂后，鳗鲡血清GtH含量可较长时间的保持较高水平，血清GtH的变化较常规注射方法缓和，注射乳剂还可使鳗鲡脑垂体GtH含量升高。     

LHRH-A、DA对施氏鲟脑垂体碎片释放GtH的影响

为了探讨在古老的软骨硬鳞鱼中促性腺激素(GtH)的双重内分泌调节作用,本实验设计用离体灌流的方法研究促黄体素释放激素类似物(LHRH-A)和多巴胺(DA)对施氏鲟脑垂体碎片分泌GtH的影响。引入10、100和1 000 nmol/L 3个浓度的LHRH-A对施氏鲟脑垂体碎片3次脉冲式刺激实验;每次间隔1 h,持续5 min,研究不同剂量LHRH-A对鲟鱼脑垂体释放GtH的作用;用200 nmol/L DA对施氏鲟脑垂体碎片持续2 h灌流后引入5 min的1 000nmol/L LHRH-A刺激实验,研究DA如何抑制鲟鱼脑垂体释放GtH。每5 min收集一管灌流液,用放射免疫测定法(RIA)检测灌流液中GtH的含量。结果显示,低剂量LHRH-A随着刺激引入脑垂体释放GtH出现波浪式的增加,中、高剂量出现释放延后现象。LHRH-A在10nmol/L到1 000 nmol/L范围内对刺激脑垂体释放GtH没有剂量依存关系。DA对施氏鲟脑垂体碎片GtH的分泌没有显著影响,但是可以抑制LHRH-A引起的GtH分泌,即DA不能抑制施氏鲟GtH的基础分泌,而只能抑制LHRH-A诱导的GtH分泌。研究结果证明,在高等硬骨鱼类中存在的双重神经内分泌调节在古老的鲟鱼中也存在。     

环境温度与鲤鱼催产效应的关系

本文叙述了用鲤鱼垂体激素催产发育成熟的雌鲤鱼，催产效应时间与水温有着密切的关系。观察了240尾催产后顺产的鲤鱼，经数理统计分析，水温(x，度)和效应时间（y^,小时）之间的关系可以用曲线回归方程式y^=-x/3.15-0.2426x表示。     

大黄鱼血清IgM纯化及其兔抗血清的制备

分别用饱和硫酸铵二次盐析法和蛋白A亲和层析法对健康大黄鱼（Pseudosciaena crocea）血清中的免疫球蛋白（IgM）进行分离纯化,所得产物用SDS-PAGE进行检测.结果表明,蛋白A亲和层析法可以较好地分离到高纯度的大黄鱼血清IgM,产物的电泳胶中只有重链和轻链2个条带;饱和硫酸铵二次盐析法除了有这2个条带,还有很多杂带,而且蛋白A亲和层析法更为简便、快速,因此用蛋白A亲和层析法分离纯化IgM优于饱和硫酸铵二次盐析法;大黄鱼免疫球蛋白重链的分子量在76 kD左右;轻链分子量在28 kD左右.用纯化的大黄鱼IgM免疫实验兔,获得效价高达1：40 960的兔抗鱼IgM血清.本实验所建立的蛋白A亲和层析法提取大黄鱼血清IgM可以方便、快捷地获得高纯度的产物,适合在实验室中纯化鱼类IgM.本研究所制备的兔抗大黄鱼IgM血清可为今后的相关研究工作打下基础.[中国水产科学,2006,13（3）：475-479]     

中华鲟、史氏鲟及达氏鳇血清免疫球蛋白的纯化及部分特性分析

采用饱和硫酸氨分步沉淀和Sephadex G200凝胶层析的方法，首次分别纯化制备了健康非免疫状态下中华鲟（Acipenser sinensis）、史氏鲟（Acipenser schrenckii）和达氏鳇（Huso dauricus）的血清免疫球蛋白（Ig） ，并采用聚丙烯酰胺凝胶电泳（PAGE）、SDS-聚丙烯酰胺凝胶电泳（SDSPAGE）、蛋白免 疫印迹（Western blotting）及免疫琼扩实验等方法对其Ig及Ig亚单位的分子量和部分特性进行了分析。PAGE及SDSPAGE的结果显示：史氏鲟，中华鲟和达氏鳇IgM的相对分子量分别为867 kD， 896 kD和924 kD；3种鲟鱼Ig的重链分子量均为88 kD，都具有29 kD的轻链，其中达氏鳇还另有一分子量约为26 kD的轻链蛋白。分子量的测定及计算结果显示鲟鱼的Ig为四聚体。Western-blotting的检测结果表明，3种鲟鱼Ig的重链与其Ig具有同样的抗原性，在硝酸纤维素膜上可被兔抗鲟Ig多克隆抗体所识别，而轻链的Western blotting检测结果则呈阴性。免疫沉淀反应的结果显示，3种鲟鱼的血清及其Ig与相互之间的兔抗Ig血清有免疫沉淀反应，但与兔抗鲤Ig血清无免疫沉淀反应，这表明3种鲟科鱼类的Ig在结构和序列上是较为相似的，而与鲤鱼等高等硬骨鱼类的Ig存在较大的差别。     

绵羊血清免疫球蛋白G的纯化及分子量的测定

采用硫酸胺分级沉淀法、葡聚糖凝胶Sephadex-25层析脱盐、DEAE-纤维素纯化等方法获得绵羊血清免疫球蛋白G。通过SDS-聚丙稀酰胺凝胶电泳法进行分离获得绵羊血清免疫球蛋白G的重链（H链）和轻链（L链），与已知分子量的标准参照蛋白比较，测得重链的分子量约为53000—57000Da，轻链的分子量约为27000—29000Da。     

钙离子浓度对日本沼虾感光细胞内可溶性Gq蛋白α亚基含量的影响

研究了钙离子浓度对日本沼虾（Macrobrachium nipponense）感光细胞中可溶性Gq蛋白α亚基含量的影响。以暗适应状态日本沼虾的复眼视网膜为材料，分别用高钙溶液、生理溶液、低钙溶液进行孵育，应用SDS—PAGE及免疫印迹技术对其可溶性蛋白粗提取液进行分离和鉴定，并利用Tanon GIS凝胶图像处理系统对蛋白条带的含量进行分析，结果显示：不同钙离子浓度中孵育的日本沼虾感光细胞蛋白的条带数量和分子量大小基本相同。兔抗Gqcx多克隆抗体在高钙溶液、生理溶液及低钙溶液处理的日本沼虾感光细胞中均识别了可溶性Gq蛋白仅亚基，分子量为42ku，可溶性Gq蛋白α亚基的含量分别是8．6％、8．4％和7．2％。在暗适应条件下，细胞中可溶性Gq蛋白α亚基的含量与钙离子浓度成正比，这表明钙离子浓度的升高可俾刍牛膳结合的Go蛋白α某转化为可溶性的Ga蛋白α亚基。     

鲻鱼的催产

亲鱼体长42-45厘米,4-5龄,11月至12月为产卵期,采用鲻鱼（Mugil ccphalus）脑垂体及西那浩灵（Synahorin）进行催产试验。当只注射1-3个垂体时未见排卵,而用1个垂体加西那浩灵20家兔单位混合,作二次肌肉注射,二次相隔时间为12小时,获得排卵,雄鱼不必注射激素。     

催产激素对尼罗罗非鱼人工繁殖效果的研究

根据催产激素注射后前期血清类固醇激素的水平变化,筛选了尼罗罗非鱼(Oreochromis niloticus)最佳催产激素组合。为了进一步验证该催产激素组合的生产效果,在尼罗罗非鱼繁殖生产期间进行人工催产试验,观察催产后144 h内实际含卵雌鱼比例与催产效应时间,并从卵子形态变化方面观察催产效应。结果显示:水温21~29℃时,催产试验组平均获产率为57.5%,对照组平均获产率为27.5%,试验组的平均获产率显著高于对照组(P0.05);试验组含卵时间集中在72 h内,对照组含卵时间比较分散;催产后72 h,试验组50%雌鱼卵子发育达到成熟期,对照组仅30%。试验结果为尼罗罗非鱼全人工繁殖生产提供了参考依据。     

中华鲟和达氏鳇血清免疫球蛋白的木瓜蛋白酶水解片段分析

首先采用饱和硫酸氨分步沉淀和Sephadex G-200凝胶层析的方法,获得了非免疫状态下中华鲟(Acipenser sinensis Gray)和达氏鳇(Huso dauricus Georgi)的血清免疫球蛋白(Ig),在此基础上使用木瓜蛋白酶水解对所获得的免疫球蛋白片段进行了酶解,并采用SDS-PAGE和Western-blot等方法分析了所获得的水解片段。结果显示,2种鲟鱼的免疫球蛋白均可被木瓜蛋白酶水解蛋白,通过Sephadex G-100凝胶层析后均可得到两个完全分离的、均一的蛋白峰。SDS-PAGE检测两个水解片段的相对分子量分别为44KD和66KD,West-ern-blot检测结果显示,66KD的片段可以在硝酸纤维素杂交膜上被各自的兔抗鲟IgM多克隆抗体所识别,而44KD片段的检测结果为阴性。这表明,2种鲟科鱼类的木瓜蛋白酶水解特性相同,提示鲟科鱼类的免疫球蛋白在免疫学及生化方面具有较大的相似性。     

运动功能相关的中华鲟和西伯利亚鲟幼鱼侧视形态比较研究

通过选取11个侧视形态特征度量进行主成分分析(principal component analysis,PCA),同时设置26个标记点进行相对扭曲分析(relative warp analysis,RWA),比较了中华鲟(Acipenser sinensis)和西伯利亚鲟(A.baerii)幼鱼在侧视形态上的差异以及相对扭曲分析和传统多元分析在鲟鱼形态研究中的优劣。结合前期对它们的有氧游泳能力比较结果(西伯利亚鲟比中华鲟强25%),分析了鲟鱼形态的水动力功能。主成分分析比较表明,中华鲟头长、头高、背鳍前基点之后的躯干高度、尾柄长、背鳍前缘长度均显著大于西伯利亚鲟(P<0.05),而尾鳍上叶显著短于西伯利亚鲟(P<0.05)。相对扭曲分析计算样本的几何信息并可视化统计结果,表明中华鲟的吻厚、吻长、头高、头长、躯干后半段高、背鳍前缘长显著大于西伯利亚鲟,而尾柄长和尾鳍上下叶长显著小于西伯利亚鲟(P<0.001)。上述形态特征直接影响着两种鲟鱼的游泳能力。     

Induced meiotic gynogenesis in shovelnose sturgeon

Gynogenesis was induced in three shovelnose sturgeon (Scaphirhynchus pIatorynchus) by heat shock after egg activation with UV-treated paddlefish (Polyodon spathula) milt. Ultraviolet dosage (J m–2) for the pooled milt samples was calculated using the following linear regression equation: Dosage = 2405.27 – 352.80X 19.78X2 (X = percent transmittance of milt). Activated eggs were incubated at 18 °C until shocking at 35 °C. Shock duration was applied at 0.050 intervals from 0.15 to 0.40 0 (8.25 to 22.00 min post-fertilization; 0 at 18 °C = 55 min). The highest yield of gynogenotes (16%) was observed at 0.25 0 for female 3, 10 % at 0.30 0 for female 2 and 12% at 0.35 0 for female 1. The percentage of viable gynogenotes responded quadratically to the tau index (s/0) when shock treatments were applied. The higher yields of viable diploid sturgeon gynogenotes were achieved when eggs were heat shocked at embryological ages ranging from 0.25 to 0.35 0 (approximately 14 to 19 min post-activation at 18 °C). No viable hybrids were produced in the control fertilization of sturgeon eggs with intact paddlefish sperm which verified the gynogenetic origin of the offspring produced. © Rapid Science Ltd. 1998     

Influence of yolk blackish pigmentation of Siberian sturgeon on reproductive performance and larval survival

Within a cohort of farmed Siberian sturgeon (Acipenser baeri), some females produce eggs which show a blackish pigmentation of the ooplasm. An experiment was carried out to test whether this characteristic might influence the reproductive success and larval survival as compared to females having usual non-pigmental yolk. There were no differences in either the progress of germinal vesicle migration or the 50% in vitro maturation time course before hormonal injection. The latency, relative quantity of eggs, fertilization rate and embryonic survival were similar. The pigmentation of yolk significantly depressed the second part of embryogenesis. Neither the number of hatchlings, normal or abnormal, survival after weaning and the mean weight of 14 days post-hatch larvae were different. For the first time, rates of hatching for normal (70%), abnormal (11%) and non-hatched (7%) larvae, referred to the embryo survival at final stages, are reported with normal appearance eggs. © Rapid Science Ltd. 1998     

可见植入荧光标记和编码金属标对达乌尔鳇标志效果的初步研究

2011年9月底至2012年5月,采用可见植入荧光标记(VIE)和编码金属标(CWT)标志全长为(28.06±1.79)cm的达乌尔鳇(Huso dauricus),检验了VIE和CWT的标志保持率及其对达乌尔鳇生长和存活的影响。结果显示,经过210 d实验,VIE标志达乌尔鳇吻部腹面,CWT标志达乌尔鳇背部前(第1～2块背骨板间下方肌肉)、中(第6～7块后背骨板间下方肌肉)、后(第11～12块背骨板间下方肌肉)的标志保持率分别为:95.83%、92.00%、92.59%、92.59%。实验中VIE出现破碎、脱落或迁移致使标志可识别性下降;CWT背部中这一标志部位操作最为简便。标志30 d,VIE标志组的保持率较CWT背部不同部位标志组及对照组低,且差异显著(P<0.05),此后31～210 d并无显著性差异。VIE和CWT背部不同部位标志对达乌尔鳇的生长无显著性差异。结果表明,VIE和CWT均可用于标志达乌尔鳇。进行短期大规模达乌尔鳇的标志放流可使用成本相对较低的VIE标志。     

饥饿期间中华鲟幼鱼血液与肝脏酶活力的变化

将中华鲟幼鱼持续饥饿43 d,每隔6 d采样1次,研究饥饿期间中华鲟幼鱼血液与肝脏7项酶活力指标的变化。结果表明,饥饿期间中华鲟幼鱼血液丙氨酸转氨酶活力波动范围为42.1~134.6 U/L,在第13天与第37天均出现显著下降;天冬氨酸转氨酶活力在饥饿19 d后出现下降,最低值与最高值分别为310.1±52.4 U/L与420.8±83.4 U/L;溶菌酶活力呈现上升趋势,饥饿43 d组与饥饿前期的4个组之间具有显著差异;总抗氧化能力在饥饿19 d后下降至较低水平,饥饿13 d组显著高于饥饿37 d组;肝脏Na+-K+-ATP酶活力在饥饿前期下降,在饥饿37 d后显著升高,与饥饿前期的4个组之间具有显著差异;乳酸脱氢酶活力在饥饿13 d后显著下降,波动范围为5.101~3.154 U/g;碱性磷酸酶活力呈现上升趋势,饥饿19~43 d的5个组显著高于饥饿1 d组。从总体上看,中华鲟幼鱼生理指标在饥饿19~25 d后产生较大变化,这可能与其新陈代谢与能量供应水平下降有关,在人工养殖与救护工作中应防止中华鲟幼鱼受到长期的饥饿胁迫。     

Verification of ploidy level in sturgeon larvae

Chromosome preparations and assay of the microsatellite locus Afu‐68 were used to determine ploidy in Siberian sturgeon (Acipenser baeri Brandt) and F₁ hybrids of Siberian sturgeon and Russian sturgeon (Acipenser gueldenstaedti Brandt). The chromosome number and microsatellite locus Afu‐68 were compared and these analyses were used for identification of ‘haploid’, ‘diploid’ and ‘triploid’ progeny of the studied cross of A. baeri× (A. baeri×A. gueldenstaedti).     

Evaluation of lethal and non-lethal sampling methods for the detection of white sturgeon iridovirus infection in white sturgeon, Acipenser transmontanus (Richardson)

Pectoral fin tissue of white sturgeon was investigated as a potential non-lethal sample source for the detection of white sturgeon iridovirus (WSIV) infection. Histopathology and polymerase chain reaction (PCR) results using fin tissue were compared with the standard lethal histopathology sampling method that utilizes head tissue. Tissues for each of the three sampling methods were collected weekly for 8 weeks from individual sturgeon undergoing an experimental cohabitation challenge with fish infected with the Abernathy isolate of WSIV. Non-lethal fin histopathological evaluation did not reveal infection during the first 3 weeks of sampling, while non-lethal PCR and the lethal method were variable. However, all three sampling methods were equally capable of identifying infection from 4 to 8 weeks post-exposure. Of the survivors tested, all were negative by PCR and the lethal method, and only one fish was identified as being positive by non-lethal fin histopathology. In another experiment, all three sampling methods were applied to asymptomatic WSIV carriers in a case study conducted at the Kootenai Tribal Sturgeon Conservation Hatchery. Results showed that both lethal and non-lethal fin histopathology were equally effective in detecting infection, but PCR was unable to identify this strain of WSIV. Depending on the virus isolate, these results suggest that non-lethal sampling of fin tissue (histopathology or PCR) is comparable with the lethal sampling method at identifying WSIV infection once infection is established, and under certain circumstances may provide an alternative to lethal sampling.     

High-energy diets for white sturgeon, Acipenser transmontanus Richardson

Four diets formulated for salmon were fed to 0.11 kg white sturgeon, Acipenser transmontanus Richardson, for 8 weeks. Dietary compositions ranged from 258 to 402 g lipid kg⁻¹, 535–378 g protein kg⁻¹ and 22.7–14.4 g protein MJ⁻¹ gross energy.
Fish in all treatments grew rapidly, utilized the diets efficiently and had body compositions similar to what has been found in previous studies, but there were some dietary effects. Sturgeon fed the diet with the highest lipid content and lowest protein/energy ratio had lower ( P < 0.05) specific growth rate, feed efficiency, and liver moisture and protein contents, and 6-phosphogluconate dehydrogenase activity, but higher liver lipid contents than fish fed the other three diets. Condition factor, organ to body weight ratios, whole-body and plasma concentrations of protein, glucose and triglyceride, and liver glucose-6-phosphate dehydrogenase, isocitrate dehydrogenase and malic enzyme activities did not differ significantly among dietary treatments. This suggests that white sturgeon subyearlings can utilize diets with high lipid contents (258–357 g kg⁻¹) to display good growth without major adverse effects on body composition and liver lipogenic enzyme activities.