黄鳝精子活力检测和精子入卵早期过程观察

采用Olympus3×51相差系统显微镜和SQIAS—1000彩色精液质量图文分析系统检测黄鳝精子活力。结果表明,在NaCl溶液浓度为0～0.3%时,黄鳝精子激活比例随溶液浓度升高而极显著增加(P<0.01);当NaCl浓度达到0.7%时,精子激活比例、直线运动速度和鞭毛摆动频率均显著(P<0.05)或极显著(P<0.01)降低。扫描电镜观察显示:黄鳝成熟卵卵壳膜上的精孔区呈漏斗状凹陷,其底部中央可见一精孔管外孔,口径约4.22±0.66μm;黄鳝精子入卵速度缓慢,受精过程较长,从精子附着于卵球表面到精孔管完全堵塞,约30s～5min。     

条纹锯鮨精子超微结构及其入卵过程的电镜观察

采用扫描和透射电镜技术对自然成熟的条纹锯精子、卵子及精子入卵过程进行观察。观察结果显示,其精子由头部、中段和尾部三部分组成:头部主要由细胞核构成,无顶体结构;中段由线粒体、中心粒复合体(近端中心粒和基体)、袖套组成;尾部主要由轴丝组成,外部包裹质膜,轴丝为典型的"9+2"结构。卵子表面分布纵横交错的网纹,均匀分布着大小不一的微小孔,在卵壳的动物极精孔区的中央有一个受精孔。在授精后10 s即可见到精子通过受精孔进入卵子,刺激卵子发生形态变化封闭受精孔,阻止其他精子入卵,60 s可见受精孔完全封闭。     

史氏鲟精子入卵过程的扫描电镜观察

史氏鲟(Acipenserschrencki)隶属鲟形目(Acipenseriformes)、鲟科(Acipenseridae)、鲟属(Acipenser),是黑龙江特产和重要的经济鱼类之一,为淡水定居种类。鲟鱼是古老的生物类型,世界上约有28种,仅分布于北半球。由于自然环境的改变和人类活动的影响,所有鲟鱼均处于不同程度的濒危状态,有的已近灭绝[1,2]。我国有8种鲟鱼,其中黑龙江2种,即史氏鲟和达氏鳇。近年来,随着人工养殖技术的提高和完善,史氏鲟成为国内最主要的鲟鱼养殖种类。有关史氏鲟人工养殖、基因组DNA含量、胚胎发育等国内有较多研究,而对受精生物学方面研究尚未见报道。特殊…     

锯缘青蟹精子入卵过程的扫描电镜观察

利用扫描电镜详细观察了锯缘青蟹精子入卵的过程。精子以其核突起附着在卵膜上,并迅速发生顶体反应。顶体反应时,顶体囊外翻,顶体管前伸,精子核辐射臂收缩,并拖至顶体囊的后部。顶体管迅速穿过卵黄膜,携带核物质一同进入卵子。锯缘青蟹为多精着卵,数精入卵。本文同时探讨了精子顶体反应机制以及受精过程卵子的作用。     

中国对虾精子入卵过程的观察

利用光镜和电镜对中国对虾精子入卵过程进行了观察。结果表明，精子的核物质籍胞吐作用穿过卵黄膜到达卵膜，其余部分则留在卵黄膜外。只有在卵子激活前到达卵膜的精子才能参与受精作用，其余的精子将被卵子激活所形成的胶质层推离卵子。中国对虾精子激活过程，没有发现顶体丝的形成。精子的入卵开始于卵子的皮层反应后期，此时精卵的接触逐渐密切，首先接触部位的卵膜稍有突起，然后开始下陷，同时这一部位的卵膜变得粗糙不平，随后卵膜解体，精子由此进入卵子。中国对虾精子的入卵位置是随机的，有多精入卵的现象。     

索氏六须鲶受精早期精子入卵的扫描电镜观察

用扫描电镜对索氏六须鲶受精早期精子入卵过程进行系统观察,结果显示,索氏六须鲶在受精后３０－６０ｓ内完成精子入卵过程,在受精后２ｍｉｎ精子开始逐渐解体,直至受精后１５ｍｉｎ左右,受精卵外精子全部解体。通过对精子和卵子的精孔管的测量判定索氏六须鲶精方式为单精受精。     

中华鲟精卵特征及精子入卵早期过程观察

中华鲟精子的头部呈短棒状,前端具顶体,尾部扁平细长;精子密度在不同个体之间差异较大,实验所获3尾雄鱼的精子密度分别为5.04×109个/mL、1.74×109个/mL、2.25×109个/mL;中华鲟卵的直径大,平均长径0.45 cm,短径0.42 cm;受精孔6~15个,排列在卵的极斑处;通过扫描电镜观察了中华鲟精卵的受精过程,发现从受精5 s开始就有精子进入受精孔,10m in后受精孔完全封闭,其间卵膜未发现有任何变化。还对中华鲟受精方式的多精入卵、单精受精的机制做了探讨。     

暗纹东方鲀精、卵超微结构及精子入卵早期电镜观察

本文对暗纹东方精、卵结构和精子早期入卵进行了电镜观察。观察结果：精子为鞭毛型结构,头部两端呈椭圆长柱形,核凹窝较深,颈部含８个线粒体,尾部微管束为“９＋２”结构,双尾丝裸露;卵为圆球形,卵膜表层有许多嵴、沟、微孔和一个受精孔;受精方式为单精或多精入孔入卵,单精受精。本文还对受精方式、人工繁殖技术等进行了讨论。     

诱导鲤雌核发育时精子入卵的扫描电镜观察

以雌核发育性成熟鲤(Cyprinuscarpio)卵子和经紫外线照射处理遗传失活的鲫鱼(Carassiusauratus)精子为材料,在再次诱导其雌核发育时,通过扫描电镜和光学显微镜对比观察诱导组和正常受精组精子入卵的情况及鲤鱼成熟卵受精过程中精孔区的外部形态变化。结果表明,无论试验组或对照组在受精后2s精子就能迅速到达精孔区。精子入孔后,受精孔便被絮状物质堵塞;受精后35～40s壳膜表面开始模糊;60s后,精孔区絮状物及精子均消失。     

暗纹东方Tui精,卵超微结构及精子入卵早期电镜观察

本文对暗纹东方Ｔｕｉ精、卵结构和精子早期入卵进行了电镜观察。观察结果：精了耿鞭毛型结构,头部两端呈椭圆长柱形,核凹窝较深,颈部含８个线粒体,尾部微管束为“９＋２”结构,双尾丝裸放为圆球形,卵膜表层有许多嵴、沟、微孔和一个受精孔;受精方式为单精或我精入孔入卵,单精受精。本文还对受精方式、人工繁殖技术等进行了讨论。     

Cryopreservation of turbot Scophthalmus maximus (L.) sperm: fertilization and hatching rates

The present paper assesses the fertilization and hatching rates of an artificial fertilization series (n=1153) using fresh and cryopreserved sperm from 49 specimens of turbot Scophthalmus maximus (L.), carried out to confirm the results of a previous study, with the ultimate aim of transferring these cryo‐preservation techniques to commercial hatcheries. No significant differences were found between the fertilization rates of the two groups (fresh and cryopreserved sperm) when their respective fertility rates were >69.2%, which was the case in 75% of all fertilizations. Likewise, no significant differences in hatching rates were found. In order to use the cryopreserved sperm more efficiently, a key concern for commercial use of this technique, we also experimented with a lower sperm:diluent ratio (1:1) than used previously at our centre. We also compared the traditional 0.5‐mL straws with 2‐mL cryotubes able to contain a higher volume of sperm, finding no significant differences in the resulting fertilization and hatching rates. In conclusion, the use of cryopreservation for turbot sperm presents major advantages for broodstock management in commercial hatcheries.     

大菱鲆受精过程的细胞学观察

采用人工授精和组织切片技术对大菱鲆（Scophthalmus maximus L．）受精过程进行细胞学观察。大菱鲆成熟卵处于第二次成熟分裂的中期。精子入卵后，卵子被激动，第二次成熟分裂继续进行，同时发生皮层反应；受精后15min出现精子星光；受精后20min雄原核早于雌原核形成，然后两性原核逐渐靠拢；受精后30min两原核相互靠拢，结合线清晰；受精后40min两原核结合线逐渐消失联合成合子核，之后合子核核膜消失；受精后50min合子核处于第一次有丝分裂中期；受精后60min第一次卵裂完成。[中国水产科学，2006，13（4）：555—560]     

Artificial fertilization in turbot, Scophthalmus maximus (L.): different methods and determination of the optimal sperm–egg ratio

With the aim of improving artificial fertilization (AF) in turbot, Scophthalmus maximus (L.), a series of fertilization experiments was carried out under dry conditions and different wet conditions (eggs/sea water: 2V/V and V/V). Another series of fertilization experiments was carried out with different quantities of sperm pool to determine the optimal ratio of spermatozoa to eggs for each AF method. Sperm pool from two males and eggs from spawns with a viability rate of > 70% were used. The sperm pool’s density (0.4–5.18 × 10⁹ sperm mL^–1) and motility (1–5) had been assessed previously. Significantly different fertilization rates were found when comparing 2V/V and V/V wet conditions. Significantly higher fertilization rates were found in dry fertilization when the sperm–egg ratio was > 9000 spermatozoa per egg and, under wet condition V/V, at 3000–4000 spermatozoa per egg.     

大菱鲆生长性状在不同生长发育阶段的相关分析

为培育优良大菱鲆养殖品种,收集来源于不同国家、不同地区和不同批次的大菱鲆亲鱼8批,从各群体中挑选性腺发育良好的个体,利用不平衡巢式设计方法和人工授精技术,每条雄鱼个体配两条雌鱼个体,共构建38个父系半同胞和76个母系全同胞,分别测定了每个家系生长到25和80日龄的体重、全长和体长,利用父系半同胞组内相关法估计各性状之间的相关性。对大菱鲆遗传相关分析表明,体重和全长的遗传相关系数为0.95,体重和体长的遗传相关系数为0.88和1.00,全长和体长的遗传相关系数为0.99。对大菱鲆表型相关分析表明,体重和全长的表型相关系数为0.881,体重和体长的表型相关系数为0.622和0.871,全长和体长的表型相关系数为0.947。并且经相关系数显著性检验,各性状间的表型相关都呈极显著水平(P<0.01)。另外,多元回归分析建立了全长(X1)和体长(X2)对体重(Y)的回归方程,生长性状与体重的复相关指数为R2=0.789,说明它们都是直接影响体重的重要指标。     

Larval growth of turbot, Scophthalmus maximus (L.) produced with fresh and cryopreserved sperm

The present paper assesses the fertilization and hatching rates, as well as the growth, of larvae obtained from four artificial fertilizations (AF) using fresh and cryopreserved sperm of the turbot Scophthalmus maximus (L.). Larvae growth in both sperm groups, measured in terms of length and weight at culture days 0, 7, 14 and 31, are compared, as well as their growth rates. The two groups' fertilization and hatching rates were not significantly different. Likewise, no significant differences in length and wet weight of 7‐ and 14‐day‐old larvae were found using fresh and cryopreserved sperm; however, significant differences were found in 31‐day‐old larvae, which were more attributable to the variability inherent in larval turbot culture, and to variability in the reproductive specimens used in our study, than to the type of sperm employed. These results indicate that the type of sperm used in artificial fertilization, i.e. fresh or cryopreserved, is not a determining factor, either for fertilization and hatching, or for subsequent larval development. Our results also confirm once again the high quality of cryopreserved turbot sperm, and its usefulness in commercial hatcheries.     

Studies on Herpesvirus scophthalmi infection of turbot Scophthalmus maximus (L.) ultrastructural observations

Abstract. Recent heavy mortalities amongst O+ group turbot at a fish farm were found to be associated with a herpes-type viral infection of the epithelia of the skin and gills. The morphology of the virus is described with ultrastructural observations on its morphogenesis and release from infected cells.     

大菱鲆胚胎玻璃化方法研究

用大菱鲆(Scophthalmus maximus)神经期胚对6种抗冻剂的毒性进行检测,发现其毒性排列为1,2-丙二醇(PG)<甲醇(MeOH)<甘油(Gly)<二甲基甲酰胺(DMF)<乙二醇(EG)<二甲亚砜(DMSO).以DMF为主因子配制和筛选出11种玻璃化程度较好的玻璃化液,并用大菱鲆肌节期胚对其中5种玻璃化程度最好的玻璃化液进行检测,结果显示,A4(DMSO 20%+DMF 25%)、A7(DMSO 25%+DMF 20%)较适合于大菱鲆胚胎的平衡处理,利用A4和A7平衡处理神经期胚、肌节期胚、心跳期胚、出膜前期胚,结果显示大菱鲆神经胚和肌节胚对玻璃化液的适应能力较强.实验测试出不同时期胚胎在两种玻璃化液中的成活率,为大菱鲆各期胚胎在玻璃化液中的平衡处理提供了依据.利用大菱鲆肌节胚对不同的平衡步骤进行了筛选,发现五步法的平衡效果较好.     

Studies on Herpesvirus scophthalmi infection of turbot Scophthalmus maximus (L.): Histopathological observations

Abstract. The histological features of the disease caused by infection of turbot by Herpesvirus scophthalmi are described. Infection of epidermal cells of skin and gill resulted in the formation of giant cells showing a variety of nuclear and cytoplasmic changes associated with the replication of the virus. The effects of the condition together with possible control measures are discussed.     

Influence of photoperiod, frequency of stripping and presence of females on sperm output in turbot, Scophthalmus maximus (L.)

Abstract. The effect of four environmental conditions was investigated upon sperm output in turbot, Scophthalmus maximus (L.), submitted to three different rhythms of stripping. Males kept under a natural light cycle and under a 6-month contracted light programme released a similar sperm output in terms of total volume of semen produced per fish during the experimental period (4·9 ± 0·9ml), mean sperm concentration (29·4 ± 2·8 × 10⁹ spermatozoa/ml) and total sperm number (163·2 ± 40·5 × 10⁹ spermatozoa). Attempts to stimulate spermiation for a second time just after the end of the natural reproduction period resulted in the release of low sperm output (total volume of semen: 1·6 ± 0·4 ml; mean sperm motility: 2 min 36s ± 0 min 47s; mean sperm concentration: 47·6 ± 10·2 × 10⁹ spermatozoa/ml; total sperm number: 84·5 ± 25·3 × 10⁹ spermatozoa). Stripping frequency had no effect on total volume of semen, mean sperm motility and total sperm number. Monthly collection did not modify sperm samples in relation to stripping rank. However, decreasing volume, motility and sperm concentration were observed when males were stripped fortnightly and weekly. During the natural spawning period, the presence of females in the tank enhanced mean sperm motility (from 3 min 27s + 0 min 52s to 6 min 38s ± 1 min 38s).     

Growth and survival of young turbot (Scophthalmus maximus L.) produced with cryopreserved sperm

This study assessed the growth and survival over a year of two groups of 4‐month‐old turbot Scophthalmus maximus (L.) derived from artificial fertilization with fresh (FG) and cryopreserved sperm (CG). Growth in both groups, measured monthly in terms of length and weight, were compared. Survival was also recorded. No significant differences were found when we compared weight and length data in both groups. Growth rates were similar between FG and CG young turbot during 1 year. Likewise, the same survival rate (92.2%) was found in both groups. Our results show the good survival and growth of young turbot obtained from cryopreserved sperm, and confirm the cryopreservation technique as a useful tool for raising turbot for commercial purposes.