Physio-Chemical Characteristics of Seminal Plasma and Development of Media and Methods for the Cryopreservation of European eel Sperm     

J. F. Asturiano  L. Pérez  D. L. Garzón  F. Marco-Jiménez  D. S. Peñaranda  J. S. Vicente  M. Jover 《Fish physiology and biochemistry》2004,30(3-4):283-293

The high sperm density, together with the short spermatozoa swimming time, makes European eel sperm manipulation and assessment for quality difficult. Two diluting media (K15 and K30) previously designed for Japanese eel sperm were tested. After 24 h, European eel sperm showed significant reduction in the percentage of motile spermatozoa after activation and different motility parameters (VAP, angular velocity; VCL, curvilinear velocity; VSL, straight line velocity; BCF, beating cross frequency), concluding that these media are not suitable to preserve the sperm of this species. After a hormonal treatment to induce spermiation, sperm volume, density and motility were recorded at weekly samplings. The variation of the osmolality (325–330 mOsm kg⁻¹), pH (8.4–8.6) and the ionic composition (concentration of Na⁺, K⁺, Mg²⁺ and Ca²⁺) of the seminal plasma were registered. Physio-chemical results were related with sperm quality throughout the treatment, to determine which must be the suitable characteristics of one extender for the sperm of this species, and to find the best conditions to obtain suitable cryopreservation media for European eel sperm. K⁺ concentration increased, while Ca²⁺ and Mg²⁺ concentrations showed a progressive reduction in correlation with the sperm quality improvement. Na⁺ showed a decreasing, but not significant tendency. P1 and P2 freezing media were designed considering the physio-chemical parameters as well as the ionic composition shown by the best quality sperm samples, and then compared with the previously described solutions, TNK and K30. Sperm quality was determined, checking the percentage of motile spermatozoa and motility parameters using computer-assisted sperm analysis (CASA) software. Samples were frozen after dilution (1:5, 1:20, 1:100) in different freezing media supplemented with 10% dimethyl sulfoxide (DMSO). After thawing, samples frozen with low dilution ratio (1:5) in TNK and P1 media showed higher, although not significant, spermatozoa survival (35.5 ± 14.5 and 36.6 ± 6.7%). The addition of l-α-phosphatidylcholine to the media seems to have a positive effect, as reported in the Japanese eel.  相似文献   

诱导栉孔扇贝雌核发育时精子入卵的扫描电镜观察   总被引：3，自引：0，他引：3       下载免费PDF全文

赵峰  杨爱国  刘志鸿  周丽青  王清印 《中国水产科学》2004,11(4):281-285

取栉孔扇贝 (Chlamysfarreri)精子在 80 0 μW/(cm2 ·s)的紫外线下辐射 ,然后与正常卵子受精 ,每隔 0 .5min取“受精卵”固定。在扫描电镜下连续观察精子的入卵过程 ,然后与正常精子的入卵过程进行比较。结果表明 ,紫外辐射后的精子可以划分为 3种类型 :I类精子 ,基本无明显变化 ,可以激发卵黄膜举起形成受精膜 ,同时诱发卵子发生皮层反应 ,在受精锥的作用下正常入卵 ,但入卵的时间较正常精子晚 ;II类精子 ,顶体膜破裂 ,顶体丝伸出 ;III类精子 ,顶体丝伸出 ,鞭毛脱落。由于II、III类精子在紫外辐射条件下诱发了顶体反应 ,融解卵膜的酶提前释放 ,因而不能入卵。精子鞭毛的脱落 ,使精子丧失了运动能力 ,是导致卵表面附着的精子数量较少的原因之一。无论是正常精子还是辐射后的精子 ,入卵后在卵表面都留下一个类似受精孔的通道。实验组中未发现有多精入卵现象  相似文献   

不同精子密度冷冻猪5mL细管精液效果比较     

李林林  张德福  马恒东 《畜禽业》2007,(6):20-21

采集上海白猪精液进行5mL细管冷冻,比较冷冻密度分别为0.5×109个/mL、0.7×109个/mL、1.0×109个/mL的冻后精子质量。结果表明:3种密度冷冻猪精子,解冻后精子质量差异不显著(P﹥0.05)。  相似文献   

Viability of Extended Goat Semen Stored at Refrigerated Condition     

Flocerfida Pagador Aquino  Krystel Grace Vergara  Lerma Cajuigan Ocampo  Eufrocina dela Pena Atabay 《农业科学与技术》2014,(2):169-176

Due to the small volume of goat semen ejaculate, just a few doses of goat semen were produced when the sperm concentration is 100 × 10^/mL. The study was aimed to determine the viability of extended goat semen at refrigerated condition at 5 ℃ using varying sperm concentrations and evaluated if sperm concentration lower than 100 × 10^6/mL would affect the motility, viability and sperm morphology at refrigerated condition. Using an artificial vagina, ejaculated goat semen was collected from goat semen donor aged 1.5 year. Physical evaluation of the collected semen showed an average volume of 0.54 mL, mean pH of 6.8 and a milky white color with thick consistency indicative of high concentration. Fresh goat semen had an initial average of 76% with an average initial sperm concentration of 128 × 10^/mL. The semen was divided into four treatments： sperm concentration of 100× 10^/mL, 75 × 10^/mL, 50 × 10^/mL and 25 × 10^/mL, and were stored at 5 ℃ for a period of 10 d. The semen evaluation was performed for each of the four treatments every other day. Results showed that the sperm concentration of spermatozoa affected the duration of storage based on the sperm motility percentage, viability and morphology of spermatozoa. The extended goat semen with sperm concentration of 25 and 50 million sperm/mL is optimal for storage within 6 d that gave satisfactory percentage motile, viable and morphologically normal spermatozoa.  相似文献   

不同冻融处理对猪精子顶体膜蛋白表达的影响     

曹立朋  孙培亮  安松文  金一 《安徽农业大学学报》2014,41(1):20

为探究不同冷冻解冻方法对猪精子损伤的影响,采用2种常用的冷冻解冻方法处理猪精子,分别对其顶体膜蛋白进行分离,进行SDS-PAGE电泳和总灰度值的检测和分析。结果表明,采用一次稀释法进行猪精液的冷冻保存较二次稀释法解冻后精子活率高,精子顶体膜蛋白组分中除125 ku和90 ku处蛋白表达水平低于二次稀释法外,120 ku、48 ku、36 ku均高于二次稀释法。可见,猪精子在受到更深层次(二次稀释法对精子冷冻解冻损伤大)的冷冻损伤时,伴随着顶体膜蛋白125 ku和90 ku表达水平的升高以及120、48及36 ku表达水平的降低。  相似文献   

Effect of different methods for the induction of spermiation on semen quality in European eel   总被引：2，自引：0，他引：2  

Juan F Asturiano  Luz Pérez  Daniel L Garzón  David S Peñaranda  Francisco Marco-Jiménez  Silvia Martínez-Llorens  Ana Tomás  & Miguel Jover 《Aquaculture Research》2005,36(15):1480-1487

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.  相似文献   

Cryopreservation of filefish (Thamnaconus septentrionalis Gunther, 1877) sperm     

Kyoung Ho Kang  Kang Hee Kho  Zong Tao Chen  Jae Min Kim  Young Hun Kim  & Zhi Feng Zhang 《Aquaculture Research》2004,35(15):1429-1433

The present study examined the possibility of long‐term storage, by cryopreservation in liquid nitrogen, of the sperm of filefish (Thamnaconus septentrionalis). Changes in motility, survival rate, ultrastructure and fertilization rate of the sperm after freezing and thawing were tested. For selection of the immobilizing solution, artificial seawater (ASW) of 250, 350 and 450 mOsmol kg^?1 were tested. Sperm motility was significantly inhibited in 350 mOsmol kg^?1 ASW, and restored entirely after 100% ASW (1200 mOsmol kg^?1) was added. Two cryoprotectants, dimethyl sulphoxide and glycerol, were employed. The sperm was diluted at the ratio of 1:6 with the extenders, and frozen at a freezing rate of ?40°C min^?1 to ?100°C after equilibration for 10 min at room temperature, followed by plunging into liquid nitrogen. The highest post‐thawed sperm motility and survival rate were obtained with 5% glycerol. Afterwards, the effect of different freezing rates was examined using 5% glycerol as a cryoprotectant, and the rate of ?30°C min^?1 to ?100°C showed the best result.  相似文献   

Australian sperm whales from different whaling stocks belong to the same population     

Joanna Day  David Power  Rosemary Gales  John Bannister  Maxine P. Piggott  Kerstin Bilgmann  Robert Harcourt  Luciano B. Beheregaray  Luciana M. Möller 《水产资源保护:海洋与淡水生态系统》2021,31(6):1452-1465

1. Understanding the factors driving population structure in marine mammals is needed to evaluate the impacts of previous exploitation, current anthropogenic threats, conservation status, and success of population recovery efforts.
2. Sperm whales are characterized by a worldwide distribution, low genetic diversity, complex patterns of social and genetic structure that differ significantly within and between ocean basins, and a long history of being commercially whaled. In Australia, sperm whales from the (International Whaling Commission assigned) southern hemisphere ‘Division 5’ stock were very heavily exploited by whaling.
3. The present study assessed the potential effects of whaling on the genetic diversity of sperm whales in Australia and the population genetic structure of these whales within a global context. A combination of historical and contemporary sperm whale samples (n = 157) were analysed across six regions, from south-eastern Australia (‘Division 6’ stock in the Pacific Ocean) to south-western Australia (‘Division 5’ stock in the Indian Ocean).
4. Sperm whales sampled from the ‘Division 5’ and ‘Division 6’ stocks belong to the same population based on nuclear and mitochondrial DNA (mtDNA) analyses. Four novel sperm whale mtDNA haplotypes were identified in animals from Australian waters. Levels of genetic diversity were low in Australian sperm whales but were similar to those previously reported for populations in the Indian and Pacific Oceans.
5. Given the genetic distinctiveness of sperm whales in Australian waters from other regions in the Pacific and Indian Oceans, and the lack of recovery in population numbers, further scientific studies are needed to increase our understanding of population dynamics and the effectiveness of threat management strategies in this species.

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Historical and contemporary habitat use of sperm whales around the Galápagos Archipelago: Implications for conservation     

Ana Eguiguren  Enrico Pirotta  Kristina Boerder  Maurício Cantor  Godfrey Merlen  Hal Whitehead 《水产资源保护:海洋与淡水生态系统》2021,31(6):1466-1481

1. Sperm whales have occupied the waters off the Galápagos Islands, Ecuador, for at least the past 200 years. During the 19^th century, they were the target of intensive whaling that severely depleted the population. In recent times, after commercial whaling ended, sperm whales in the region remain vulnerable to multiple threats, especially potential entanglement in fishing gear, which may hinder their ability to recover from the whaling era.
2. As a highly mobile, long-lived species, long-term analysis of the habitat use of sperm whales is necessary to establish effective conservation and management strategies. Here, contemporary (1985–2014) and historical (1830–1850) sperm whale habitat use off the Galápagos Islands was analysed and contrasted to the extent of the Galápagos Marine Reserve (GMR). Contemporary habitat use and its variability over time were modelled as a function of geographic, oceanographic, and topographic variables using generalized additive models.
3. The fine-scale habitat (<50 km) used by sperm whales was associated with topographic (i.e. depth and slope) and oceanographic characteristics (i.e. relative sea surface temperature and standard deviation of sea surface temperature), but these preferences varied over time.
4. While historical and contemporary data indicate that sperm whale habitat primarily occurred within the boundaries of the GMR, in recent years, whales were found up to 30.1% of the time outside the GMR, potentially overlapping with commercial fisheries operating in the area.
5. The dynamic nature of the relationship of this nomadic species with its habitat highlights the need of large-scale conservation efforts across the Eastern Tropical Pacific region, including the wide-scale enforcement of regulations requiring the use of Automatic Identification System in fishing vessels, the promotion of on-board fisheries observer programmes, the development of adaptive management strategies, and international collaboration to identify and mitigate threats.

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Binding characteristics and regulation of the 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) receptor on testicular and sperm plasma membranes of spotted seatrout (Cynoscion nebulosus)     

P. Thomas  D. Breckenridge-Miller  C. Detweiler 《Fish physiology and biochemistry》1997,17(1-6):109-116

The binding characteristics of 17,20,21-trihydroxy-4-pregnen-3-one (20-S) to plasma membranes prepared from the testes and sperm of spotted seatrout (Cynoscion nebulosus) were investigated using a filtration method to retain the bound 20-S. A single class of high affinity (K_d = 17.9 nM), low capacity (B_max = 0.072 nM g^-1 testes) binding sites was identified by saturation and Scatchard analyses on testicular membranes of spermiating spotted seatrout. A corresponding receptor (K_d = 22.17 nM, B_max = 0.00261 nM ml^-1 milt) was also detected in spermatozoan membrane preparations. The rates of 20-S association and dissociation were rapid, both had T_halfs of less than 1 min. Competition studies indicated that the receptor was highly specific for 20-S. 17,20-dihydroxy-4-pregnen-3-one, which had the highest affinity of the other steroids tested, had a relative binding affinity (RBA) of 14.3%. Progesterone, 11-deoxycortisol and testosterone competed with an order of magnitude less affinity (RBA's of 7.4, 1.8 and 1.1%, respectively). Estradiol displayed low affinity for the receptor (RBA = 0.4%) and cortisol did not cause any displacement at 1000-fold excess concentration. Specific 20-S receptor binding was detected in plasma membranes from testes of both spermiating and non-spermiating seatrout and on spermatozoa. Prolonged incubation of testicular fragments from a spermiating fish with gonadotropin (15 IU ml^-1 human chorionic gonadotropin) or forskolin (10 µM) caused a 2–3 fold increase in membrane receptor binding. Previous studies have shown that gonadotropin-induced upregulation of the 20-S plasma membrane receptor in seatrout ovaries is required for the oocytes to become responsive to 20-S and undergo final maturation. The existence of a 20-S membrane receptor on sperm and its upregulation in the testes by gonadotropin raises the possibility that final maturation of spermatozoa in male seatrout may be regulated by a similar mechanism.  相似文献