人工养殖西伯利亚鲟精子超低温冷冻保存研究

研究了人工养殖西伯利亚鲟精子的生物学特征及超低温冷冻保存方法。西伯利亚鲟的产精量为113.67±39.86 ml,精子密度为(6.49±3.10)×108/ml,精子活力为(85.4±9.5)%,精子寿命为353±23 s。精子密度与精子快速运动时间、精子寿命之间均存在线性相关,用方程分别表示为:y=1.0384x+1.5089(R2=0.7325);y=2.9069x+74.289(R2=0.6967)。结果表明精子密度可作为一项精子质量评价的标准。通过比较西伯利亚鲟精子在不同稀释液、不同抗冻剂和抗冻剂浓度、降温速率、解冻温度下的保存效果,结果表明:配方2作为稀释液,18%甲醇作为抗冻剂,二步法超低温(-196℃)冷冻保存精子,40℃水浴解冻取得最好的冻后活力,解冻后活力为(51.8±5.8)%。西伯利亚鲟授精的最佳精卵比为106∶1。在此精卵比下用冻精授精分别得到了(72.3±3)%的受精率和(52.9±4.1)%的孵化率,其中受精率与鲜精没有显著性差异,孵化率与鲜精有显著差异(P<0.05)。     

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。     

计算机辅助分析冷冻前后褐牙鲆精子运动特征

观察了褐牙鲆(Paralichthys olivaceus)精子在室温和低温下的活力与寿命,并应用计算机辅助精子分析系统(CASA)对超低温冷冻前后褐牙鲆精子的运动特征进行了分析,结果表明:褐牙鲆精子在室温(25℃)下,可存活4 d,在低温(4℃)下可存活7 d;鲜精的活力为(87. 74±5. 47)%,解冻后,精子的最高活性为(84. 00±3. 67)%;激活0. 5 min时,冻精与鲜精的运动精子占总精子数的百分率(MOT)无显著性差异(P>0. 05),但精子平均曲线运动速度(VCL)、平均直线运动速度(VSL)、平均路径运动速度(VAP)和精子运动路线的曲折程度(LIN)都有显著性差异(P <0. 05);激活4min和10min时,冻精与鲜精的MOT、VCL、VSL、VAP和LIN间都有显著性差异(P <0. 05)。鲜精激活0. 5 min后,直线运动、曲线运动、左右摆动和不运动的精子数目占总精子数的百分比分别为(24. 49±3. 87)%、(48. 53±4. 55)%、(24. 72±2. 86)%和(2. 27±1. 22)%;冻精激活0. 5min后,直线运动、曲线运动、左右摆动和不运动的精子数目占总精子数的百分比分别为(18. 58±1. 33)%、(35. 67±3. 00)%、(35. 24±2. 67)%和(10. 51±1. 33)%。随着激活时间的延长,褐牙鲆鲜精和冻精的运动状态均发生了改变,直线运动和曲线运动的精子数目逐渐减少,而不运动和左右摆动的精子数目逐渐增加。     

黄颡鱼精子低温保存方法的初步研究及应用

以精子活力为标准,通过对4种保存液下黄颡鱼精子保存效果对比,筛选出保存效果较佳的基础保存液,并在此基础上对不同温度(0 ℃和4 ℃)、不同青霉素浓度和添加抗冻剂与否的条件下保存液保存效果做对比研究.试验结果表明,渔用任氏液保存效果较佳,提高青霉素浓度(通常为0.5×104 ～1.0×104 IU /ml,使用2.0×104 IU /ml),添加抗冻剂(6%DMSO),在添加抗冻剂条件下降低温度(由4 ℃降为 0 ℃)均可明显提高保存效果,在最优条件(0 ℃、2.0×104 IU青霉素/ml、6%DMSO)下,12 d内能保持80%的精子活性.人工授精试验证明了经保存的精子能正常用于人工繁殖,且授精效果与新鲜精液精子无显著差异.     

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。     

环境因子对大黄鱼精子活力的影响

通过观测精子的激活率、活动时间和寿命研究了几种环境因子的变化对养殖大黄鱼精子活力的影响。试验结果表明:精子平均密度为(1.17±0.09)×1010.ml-1,盐度对大黄鱼精子活力影响较大。当海水盐度适宜(19.61~24.87)时,精子的激活率≥90%,活动时间≥9.65 min,寿命≥13.50 min;在pH=4.0~10.0的海水中,精子都能被正常激活(≥70%),适宜的pH值为7.5~8.0;不同浓度的葡萄糖、NaCl和KCl溶液对精子活力的影响不同,不同浓度的EDTA-2Na溶液均不能激活精子;无Ca2 、Mg2 或HCO3-的人工海水对精子的激活率均高达90%,但精子的活动时间却有较大幅度的缩减。     

分光光度法测定两种蟹类精子密度的研究

蟹类精子计数对其冷冻保存及种群生殖能力的探究有重要意义。为了提高计数蟹类精子数量的速度,建立了分光光度法测定蟹类精子密度的方法。比较了两种蟹类在不同波长（380nm、520nm、760nm）下吸光度（A）与精子密度（C）的关系。结果表明,在520nm波段下三疣梭子蟹（Portunus trituberculatus）精子密度和吸光度之间也存在线性回归关系,其回归方程是：A=0.058×C（R2=0.9928）;中华绒螯蟹（Eriocheir sinensis）精子吸光度与精子密度呈线性回归关系,其回归方程是：A=0.061×C（R2=0.9892）。统计检验表明,使用该方法测定蟹类精子数目,是一种快速而准确的方法,可以加速蟹类精子计数速度。     

稀有(鱼句)鲫精子主要生物学特性及活力的观察

研究了不同水体、盐度(NaCl溶液)和pH对稀有(鱼句)鲫(Gobiocypris rarus)精子活力的影响,并测定了精子的大小、密度.结果显示:稀有(鱼句)鲫精子头部长(1.246±0.083)μm,宽(1.053±0.172)μm,尾长约(37.21±2.536)μm;精子密度为(4.623±0.170)×10 ...     

条纹锯精液超低温冷冻保存研究

为建立条纹锯精液超低温冷冻保存方法,实验采用计算机辅助精子分析系统（CASA）分析了采用6种抗冻保护剂（GLY[甘油]、DMSO[二甲基亚砜]、PG[丙二醇]、EG[乙二醇]、METH[甲醇]、DMA[二甲基乙酰胺]）在4种浓度（5%、10%、15%、20%,v/v）下对条纹锯精液的冷冻保存效果。结果发现,以HBSS为稀释液,采用程序降温仪分步降温冷冻保存条纹锯精液,37℃水浴解冻后的精子中,15% PG 作为抗冻保护剂的精子运动率最高,达到（93.1±0.9）%,与鲜精差异不显著（P>0.05）,15% PG 作为抗冻保护剂的精子水浴解冻后精子的运动速度最高,平均直线速度、平均曲线速度、平均路径速度分别达到了（88.3±0.3）μm/s、（76.2±0.5） μm/s、（86.7±0.7） μm/s,与鲜精差异不显著（P>0.05）。在不同种类及不同浓度抗冻保护剂保护下,15% PG 作为抗冻保护剂的精子解冻后 1 min内运动率变化与鲜精差异不显著（P>0.05）。研究表明,15% PG为条纹锯最佳抗冻保护剂,可用于条纹锯精液的超低温冷冻保存。     

大鳍鳠精子生物学特性及几种环境因子对其活力的影响

为探究大鳍鳠(Mystusmacropterus)精子生物学特性及环境因子对其精子活力的影响,提高精子活力,本研究对其精液pH、渗透压、精子密度、精子运动参数进行测定,对精子的结构进行观察,同时设定pH、葡萄糖和离子(NaCl、KCl、CaCl₂)浓度梯度,应用计算机辅助精子分析系统(CASA)观察其对精子活力的影响。结果表明,大鳍鳠精子由头部、颈部和尾部组成,无顶体,有侧鳍。大鳍鳠平均精子密度为2.50×10⁹个/mL,精液pH为7.0～7.2,精浆渗透压为(634.16±6.66) kPa。精浆中离子成分以Na⁺含量最高,其次是K⁺,之后依次为Mg²⁺、Ca²⁺、Fe³⁺、Zn²⁺,未检测出Cu²⁺。精浆水解氨基酸总量为169239.21μmol/L,其中以亮氨酸含量最高,蛋氨酸含量最低。大鳍鳠精子经去离子水激活后,运动率、快速运动时间和寿命分别为(48.61±14.85)%、(34.0...     

Seasonal Changes in the Milt Quality of Waigieu Seaperch,Psammoperca waigiensis: Implications for Artificial Propagation

Milt quality of Waigieu seaperch was sampled at the beginning, the middle, and the end of the spawning season to assess if the sampling season has an effect on milt quality parameters. The milt volume and total sperm production were higher at the middle of the spawning season while the sperm concentration in milt was significantly higher at the beginning and the end of the spawning season. Sperm morphology and the major parameters of seminal plasma (pH, total protein, Mg²⁺, or Ca²⁺ concentrations) did not differ throughout the spawning season. The concentrations of Na⁺ and the osmolality increased throughout the spawning season. The percentage of motile cells did not differ during the spawning season while the duration of sperm motility (swimming duration) and velocity (swimming speed) were significantly different during the spawning season. The fertilization rate (75.7 ± 6.8%) and hatching rate (56.5 ± 3.1%) at the middle of the spawning season were higher than at the beginning and end periods. Results indicated that milt quality parameters and fertility success in the middle of the spawning season were higher compared to earlier and later sampling dates. Thus, we recommend collecting milt during this time for maximal fertilization and hatching success.     

Determination of spermatological properties of male Liza abu (Heckel, 1843) in Atatürk Dam Lake, Şanlıurfa

The aim of this study was to determine the spermatological characteristics in male L. abu during the spawning season. Semen was collected weekly by abdominal massage from 26 males in March. In collected semen, volume, motility, duration of motility, concentration and pH were determined. In the L. abu sperm, volume (μl), motility (%), duration of motility (s), concentration (×10⁹/ml), and pH values were found 45.76 ± 3.55, 54.25 ± 2.93, 330.15 ± 37.92, 4.27 ± 0.40 and 7.87 ± 0.05, respectively. A correlation was found between semen volume and semen pH. Semen volume and the duration of sperm motility were higher in the 2nd and 3rd sampling dates than in the 1st and 4th sampling dates (P < 0.05; P < 0.01, respectively). Neither sperm motility nor sperm concentration was affected by sampling dates. Major changes in semen pH were observed in the 4th sampling date (P < 0.001). The Pearson correlation test presented significant relationships with the duration of motility, semen volume, and motility. Semen pH values were significantly correlated with the sperm concentration and semen volume. Sperm concentration was inversely correlated with semen volume. Sperm motility and duration significantly correlated with total weight. Total length significantly correlated with the duration of motility and total weight. In conclusion, these characteristics represent a valuable baseline dataset for establishing a semen quality standard and provide background information that may be useful for assisted breeding programs in this species.     

Changes of sperm morphology,volume, density,and motility parameters in northern pike during the spawning period

Sexually mature males (BW?=?1600?±?150 g and TL?=?235?±?30 mm) of northern pike (Esox lucius L.) were randomly selected from a pond to record changes in their sperm quality parameters (spermatozoa morphology, sperm volume, density, and motility parameters) during the spawning season. The morphological and motility parameters changed significantly during the reproductive season with following trends. Only, head width was not changed during the spawning season. The longest spermatozoa and its flagellar length were found at the middle of spawning period (TL?=?38.24?±?0.37 μm and 35.14?±?0.26 μm) and shortest at the beginning of spawning period (TL?=?34.81?±?0.29 μm and 32.53?±?0.18 μm). Other morphological characters were always the lowest at the beginning of spawning period. Sperm volume was changed from 0.33?±?0.3 ml in February, 0.43?±?0.2 ml in March to 0.24?±?0.1 ml in April, and density from 16.2?±?0.2?×?109 spermatozoa ml^?1 in February, 19.4?±?0.2?×?109 spermatozoa ml^?1 in March to 4.8?±?0.2?×?109 spermatozoa ml^?1 in April. Same sperm velocity was observed in all spawning terms at 10 and 20 s after activation. Higher velocity was found at 30 and 40 s after activation in sperm collected at the middle and the end of spawning period. Significantly, higher percentage of motile sperm was observed at 20, 30, and 40 s after activation in sperm sampled at the end of spawning period. This study supports the hypothesis that longer spermatozoa swim faster.     

不同盐度激活液、K+浓度及保存时间对日本鳗鲡精子活力的影响

在6种不同盐度(34、32、30、28、26和22)激活液、3种不同K+浓度(25 mmol/L、30 mmol/L和35 mmol/L)稀释液和不同保存时间(0 h、24 h、48 h、72 h和96 h)条件下,对日本鳗鲡(Anguilla japonica)精子的活力进行观察和测定.结果表明,激活液盐度为30时精...     

Sperm quality in male <Emphasis Type="Italic">Barbus barbus</Emphasis> L. fed different diets during the spawning season

Sperm quality of Barbus barbus L. was compared among the three following dietary regimes: Group A, fed 100% commercial diet (Karpico™ containing 33% crude protein and 6% fat), Group B, fed 78% commercial diet and 22% frozen chironomid (Chironomus plumosus) larvae, and Group C, fed 56% commercial diet and 44% frozen chironomid larvae. Concentrations of polyunsaturated fatty acids (PUFAs) in Group A, B, and C were 39.1, 42.0, and 44.6, respectively, as a percentage of total fatty acids. Sperm morphology, volume, concentration and motility, total number of spermatozoa, and osmolality of the seminal plasma were compared during the spawning season. Dietary regime did not influence sperm volume, concentration, or total number of spermatozoa, osmolality of seminal plasma, or the percentage of motile sperm, but significantly affected sperm morphology (except for anterior and posterior parts of the midpiece) and sperm velocity (P < 0.05). Groups B and C showed similar sperm characteristics during the spawning season compared to Group A. Almost all parameters changed either among or within groups during the spawning season, suggesting differences in terms of the optimal time for sperm collection. The best time for sperm collection was March for Group A, but April for Groups B and C, when the osmolality of the seminal plasma measured 289 mOsmol kg⁻¹ and sperm motility was maximal. Spermatogenesis, hydration, and cell decomposition were confirmed as the three major parameters controlling sperm characteristics during the spawning season. The possible correlation between sperm morphology and motility requires further study.     

Sperm quality and cryopreservation of Brazilian freshwater fish species: a review

The Brazilian freshwater fish diversity is the richest in the world. Only 0.7% of all Brazilian species have had any aspect of their sperm biology addressed up to this date. The majority of the fish species described in this review migrate during the spawning season (a phenomenon known as piracema). Urbanization, pollution, hydroelectric dams and deforestation are some of the causes of stock depletion or even local extinction of some of these species. The knowledge concerning sperm quality and minimum sperm:egg ratio is important to maximize the use of males without reducing hatching rates. Furthermore, sperm cryopreservation and gene banking can guarantee the conservation of genetic diversity and development of adequate breeding programs of native fish species. In this review, we present and evaluate the existing information on Brazilian fish species that have been subject to sperm quality and cryopreservation studies. The following parameters were evaluated: volume of extractable sperm, sperm motility, sperm concentration, freezing media, freezing methods, and post-thaw sperm quality. Although the existing protocols yield relatively high post-thaw motility and fertilization rates, the use of cryopreserved sperm in routine hatchery production is still limited in Brazil.     

Gonadotropin releasing hormone‐analogue (GnRHa) treatment improves the milt production and sperm motility of endangered Caspian brown trout,Salmo trutta caspius,over the course of a spawning season

To determine the effect of gonadotropin releasing hormone‐analogue (GnRHa) treatment on the milt quality of endangered Caspian brown trout, Salmo trutta caspius, the sperm motility (percentage and duration of motility), sperm production (sperm density, spermatocrit and milt volume) and milt pH were measured for GnRHa‐treated (the treatment group) and untreated groups (the control group) during the spawning season. For untreated brooders, the values of the motility per cent, sperm density and spermatocrit decreased continuously during the spawning season while the milt volume, duration of motility and milt pH showed only a significant decrease at the end of the season. For GnRHa‐treated males, these parameters increased 14 days after GnRHa treatment (first milt collection) and then decreased continuously towards the end of the season. In addition, the values of milt and sperm density yielded per treated male were higher than that in the untreated group, although these were not statistically different. In any case, the total sum of yielded milt from the treatment group over the spawning season was higher than that in the untreated group. In this experiment, significant positive correlations were found between milt parameters as follows: sperm motility vs. milt pH; sperm density vs. spermatocrit; milt volume vs. spermatocrit; and milt volume vs. sperm density. The results show that the treatment of Caspian brown trout by GnRHa can improve the milt quality in terms of sperm motility and sperm production during a spawning season.     

Sperm output in rainbow trout (Salmo gairdneri) — Effect of age,timing and frequency of stripping and presence of females

An experiment was conducted to investigate the sperm production of rainbow trout. Eighty-four first-spawners were allocated randomly to three groups to be stripped at weekly, 2-weekly or 4-weekly intervals, respectively. Half of each group were exposed to females. The trial extended over two successive spawning seasons.First-season spawners were modest sperm producers. Depending on the collection frequency they produced mean stripped semen volumes of 1.17 to 1.48 ml containing 7.7 × 10⁹ to 16.8 × 10⁹ sperm. The corresponding means for the second spawning season were 3.54 to 4.59 ml and 22.2 to 35.4 × 10⁹. The spawning season extended from October to April. Production and motility increased from the beginning to mid-season, followed by a gradual decline. The effect of the presence of females was not very evident. With increased frequency of stripping, more sperm/male were obtained. Total semen volume and number of sperm/spawner obtained during the first season with weekly, 2-weekly or 4-weekly collection amounted to 24.6 ml and 160.9 × 10⁹ sperm, 13.4 ml and 135.8 × 10⁹ sperm and 8.9 ml and 101.1 × 10⁹ sperm, respectively. The corresponding values for the second season were 88.9 ml and 546.1 × 10⁹ sperm, 50.3 ml and 377.5 × 10⁹ sperm and 32.1 ml and 247.9 × 10⁹ sperm.From a practical point of view, weekly collection is feasible in second-season spawners, but a maximum of one stripping every 2 weeks appears appropriate for first-season spawners.     

Changes in Sperm Production,Sperm Motility,and Composition of Seminal Fluid in Caspian Brown Trout,Salmo trutta caspius,Over the Course of a Spawning Season

This study was carried out to evaluate milt quality in male Caspian brown trout (Salmo trutta caspius) over the course of the winter spawning season. Milt samples were collected biweekly during December and January. Chemical composition of seminal fluid, sperm production (milt volume, sperm density, spermatocrit,) and sperm motility characteristics (percentage and duration of motility) were measured. Milt volume, sperm density, osmolality, seminal minerals (Ca²⁺, Mg²⁺, K⁺, Na⁺, Cl^?), and total protein gradually decreased over the spawning season. Glucose and triglyceride content of milt did not show significant changes over the spawning season. Milt pH and the percentage and duration of motility were comparatively stable, declining only at the end of the season. Significant positive correlations were found between sperm density and seminal minerals, total protein and spermatocrit; percentage of motile spermatozoa and seminal minerals, total protein; and duration of motility and K⁺, Cl^?, total protein, and pH. Results show that season has a significant influence on milt quality in male Caspian brown trout, with the best milt being available at the beginning of spawning season.     

Effect of different methods for the induction of spermiation on semen quality in European eel

Five hormonal treatments with human chorionic gonadotropin (hCG) were tested for the induction of maturation and spermiation in male farmed eels. The main aim was to optimize previously used hormonal treatments to achieve shorter induction treatments, longer spermiation periods and/or higher sperm quality. Fish treated for just 3 weeks (treatment E) or until the onset of spermiation (treatment C) showed the worst results, while the treatment consisting of weekly administration of 1.5 IU hCG g^?1 fish (treatment A) induced the highest percentage of spermiating males, the highest number of sperm samples and sperm volumes and densities similar to the rest of the treatments (B: half hormone dosage, or D: biweekly administration). Evaluation of the sperm quality was performed by computer‐assisted sperm analysis (CASA), considering the percentage of total motile spermatozoa, the percentage of fast and medium‐velocity spermatozoa, as well as different motility parameters. Sperm samples from A‐D groups showed between 44% and 54% motile spermatozoa, and between 10% and 15% fast spermatozoa, while samples from E‐treated males showed 0% motile cells. No significant differences were found in the spermatozoa straight line velocity (VSL), curvilinear velocity (VCL) or the angular velocity (VAP), neither spermatozoa beating cross frequency (BCF) between A–D groups.