共查询到20条相似文献，搜索用时 135 毫秒
1.
《水产学报》2014,(10)
为建立条纹锯鮨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)下对条纹锯鮨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鮨精液,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(P0.05),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(P0.05)。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(P0.05)。研究表明,15%PG为条纹锯鮨最佳抗冻保护剂,可用于条纹锯鮨精液的超低温冷冻保存。 相似文献
2.
人工养殖西伯利亚鲟精子超低温冷冻保存研究 总被引：7，自引：6，他引：7
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)。 相似文献
3.
为研究适合珍珠龙精子的短期超低温保存方法,比较了不同保存液、保护剂、降温方式、解冻方式对珍珠龙(Scleropages jardini)精液超低温保存后精子活力的影响,初步解释了经超低温短期保存后珍珠龙精子活力明显较鲜精低的原因.结果显示:采用Ringer's液作为保存液,以浓度为16％的二甲亚砜作为保护剂,选取五步法超低温冷冻保存精子[精子缓慢降温至-150℃→液氮面上3 cm(约-170℃),停留4min→液氮面(约-190℃),停留1 min→液氮],40℃水浴解冻,可取得最好的冻后活力,解冻后精子的活力为(36.7±4.1)％. 相似文献
4.
【目的】本文旨在探究长期超低温冷冻保存中鞍带石斑鱼精子质膜、活力、超微结构及酶活性的变化，为阐明影响鞍带石斑鱼精子冷冻保存质量的相关机制提供理论依据。【方法】采集2022年鞍带石斑鱼鲜精及储存时间分别为23、49、61个月冷冻保存精液，用伊红-苯胺黑染色方法检测精子质膜完整性；用计算机辅助精子分析仪（CASA）检测精子运动参数；测量精浆和精子中琥珀酸脱氢酶（SDH）、过氧化氢酶（CAT）、谷胱甘肽还原酶（GR）、总超氧化物歧化酶（T-SOD）、谷胱甘肽过氧化物酶（GSH-PX）和肌酸激酶（CK）共六种酶活性的变化及三磷酸腺苷（ATP）浓度变化；用扫描电镜和透射电镜观察鲜精和冻精超微结构。【结果】伊红-苯胺黑染色检测结果发现，鲜精质膜完整性最高为83.43±2.73 %，经过超低温冷冻后，精子质膜完整性显著降低（P<0.05），且随着冷冻保存时间的延长而逐渐降低。CASA结果显示鲜精活力最高为90.47±3.34 %，经过超低温冷冻后精子活力显著降低（P<0.05），但长期保存23-61个月精子活力无显著性差异，精子活力保持在63.95±3.66 %-68.58±2.73 %，具有稳定的活力，且鲜精与冻精之间精子平均直线运动速度（VSL）、平均曲线运动速度（VCL）和平均路径速度（VAP）均没有显著差异（P>0.05）。精子超微结构显示，鲜精形态结构正常、线粒体排列结构规则、形态大小正常。经过超低温冷冻保存后，精子形态结构损伤明显，表现为精子头部质膜破损、细胞质外漏、细胞核膜破损、尾部鞭毛断裂或脱落等损伤；鞍带石斑鱼精浆和精子超低温冷冻前后六种酶活性的变化及ATP含量结果显示，经过超低温冷冻后，精子内SOD、GSH-PX和CAT三种酶及ATP含量均有显著性降低（P<0.05）。精浆中酶活力升高，除GR和CAT外，其余酶活性均差异显著（P<0.05）。【结论】长期超低温冷冻对鞍带石斑鱼精浆和精子酶活性、精子活力及精子超微结构均具有较显著影响，【意义】研究结果为鱼类精子冷冻损伤机理研究积累了丰富的数据，为鱼类精子长期冷冻保存提供了技术参考和评价指标。 相似文献
5.
以云纹石斑鱼精液为实验材料,对精子稀释液、抗冻剂种类和适宜浓度、冷冻保存液进行了筛选。结果表明,利用9g/L NaCl、10g/L KHCO3和10%小牛血清配制而成的稀释液EM1-2适宜于云纹石斑鱼精子冷冻保存,以2ml冷冻管为精子容器,在60L液氮生物保存罐中冷冻保存精子,冷冻解冻精子活力可达56.67%±5.77%,要优于TS-2、ES1-3和其他EM系列稀释液冷冻保存精子活力。利用EM1-2为基础液对抗冻保护剂进行筛选,结果显示,10%~20%的二甲基亚砜(DMSO)和1-2-丙二醇(PG)冷冻保存后精子活力无显著差异(P0.05),其中15%的DMSO和10%PG冷冻保存精子效果最优,解冻后精子活力分别可达54.52%±7.81%和57.24%±3.69%。利用冷冻保存1年的精液与云纹石斑鱼卵进行受精,受精率和孵化率均达到80%以上,与新鲜精子无显著性差异(P0.05)。本研究表明,利用EM1-2配制15%的DMSO或10%的PG可用于冷冻保存云纹石斑鱼精液。在此基础上,建立了精子冷冻库,保存精子130ml,为人工繁育和杂交育种提供了丰富的精子源。 相似文献
6.
7.
日本黄姑鱼精子生理特性及超低温冷冻保存研究 总被引：5，自引：1，他引：4
对日本黄姑鱼精子部分生理特性及其超低温冷冻保存技术进行了研究。结果表明,日本黄姑鱼精液pH值为7.0~7.5,精子密度为(11.23±2.78)×109/m l,精子寿命(229.33±17.16)s。在盐度为30~35,pH为7.5~8.5时,精子的活力最高,分别达到(88.33±2.89)%和(88±4.33)%。精子在室温(25℃)条件下,可存活24 h;在低温(4℃)条件下可以存活36 h。以D液作为稀释液,20%的乙二醇作为抗冻剂,利用快速降温法对精子进行超低温冷冻保存,38℃水浴快速解冻,解冻后的精子获得最高的活力为(47±5.78)%。 相似文献
8.
对瓦氏黄颡鱼(Pelteobagrus vachelli)精子在不同盐度和pH下的精子活力进行观察,同时研究了精子在4种不同稀释液与2种不同浓度抗冻剂组成的保存液中的超低温冷冻保存,并开展了冻精的授精实验。结果表明,瓦氏黄颡鱼精子浓度为(2.035±0.179)×1012cell·mL-1,在盐度为5.8、pH为7.17时,精子的活力都高达95%。以A液作为稀释液、10%甲醇作为抗冻剂时,冷冻保存精子效果最好,解冻后精子活力为(81.7±0.9)%。用解冻后的精子进行人工授精,获得的受精率为(88.4±2.1)%,孵化率为(74.0±0.8)%;而鲜精受精率为(91.0±0.8)%,孵化率(82±1.6)%,冻精与鲜精均无显著性差异。人工授精实验证明了解冻后的精子能正常用于该鱼的人工繁殖。 相似文献
9.
10.
为建立圆口铜鱼(Coreius guichenoti)精子超低温冷冻保存方法,采用计算机辅助精子分析系统(CASA)评价了4种稀释液(D15、D20、L1、D1), 3种抗冻保护剂[二甲基甲酰胺(DMF)、二甲基亚砜(DMSO)、甲醇(METH)]在3种体积浓度(7.5%、10%、12.5%, V/V)下对圆口铜鱼精液的冷冻保存效果,并比较了150 mOsm/kg NaCl与超纯水作为激活剂对精子活力的影响。结果表明,D1组稀释液的精子活率(MOT)最高,为(74.64±13.17)%,与鲜精无显著差异(P0.05);以10%甲醇作为抗冻保护剂的圆口铜鱼精子经超纯水激活后,测得MOT最高,达到(78.11±14.74)%,与鲜精无显著差异(P0.05),精子运动速度达最大,平均曲线运动速度(VCL)、平均直线运动速度(VSL)、VAP(平均路径运动速度)分别达到(50.28±12.46)μm/s、(35.06±10.82)μm/s、(39.44±12.46)μm/s,精子快速运动时间和寿命分别为(8.67±1.15) s、(33.33±5.00) s,显著低于鲜精(P0.05); 7.5%甲醇组和7.5%二甲基甲酰胺组的MOT次之,分别为(77.71±17.74)%、(76.42±12.49)%,抗冻剂二甲基亚砜组的MOT显著低于甲醇组、二甲基甲酰胺组(P0.05)。12.5%的3种抗冻保护剂中,几乎无精子存活;在不同种类不同浓度的抗冻剂保护下,10%甲醇组,相较于使用超纯水激活,用150 mOsm/kg NaCl激活后的精子活率和寿命更高,说明Na~+有延长冻精寿命,提高精子活率的作用。研究表明, D1+10%METH (7.8 g/L NaCl+0.5 g/L KCl+15 g/L葡萄糖+10%METH)可用于圆口铜鱼精液超低温冷冻保存,为圆口铜鱼的繁育工作和种质资源保护奠定了基础。 相似文献
11.
不同冷冻保护剂在猪精液冷冻中的作用分析 总被引：6，自引：0，他引：6
本实验采用一定浓度的甘油、EG、DMA、DMSO及其两两组合物作为冷冻保护剂。用含冷冻保护制的稀释液将精液稀释后,常温保存,观察精子活率,比较精子生存指数。结果表明：在常温下对精子毒害作用最大的是DMSO,其次是甘油,EG和DMA对精子毒害作用最小。以一定浓度的10种冷冻保护剂将对精液冷冻后观察解冻活率,发现EG和DMA混合保护剂解冻后精子活率最高。 相似文献
12.
13.
Effect of water temperature on the physiology of fish spermatozoon function: a brief review 
下载免费PDF全文
下载免费PDF全文 Hadiseh Dadras Borys Dzyuba Jacky Cosson Amin Golpour Mohammad Abdul Momin Siddique Otomar Linhart 《Aquaculture Research》2017,48(3):729-740
Motility is a key factor in function of the spermatozoon and determines semen quality and fertilizing capacity. Effective motility occurs when sperm is diluted in a swimming solution, the adequacy of which is determined by factors varying according to fish species. Spermatozoon motility rate and velocity, as well as duration of the motility period, are influenced by the temperature of the water in which broodfish are held. Increase in temperature of swimming medium beyond the optimal increases cell metabolism, leading to an increase in velocity with rapid depletion of energy resources, promoting early cessation of movement. The aim of this review was to discuss current information on the influence of temperature on quantitative spermatozoon properties, which could affect sperm function. Our findings provide a greater understanding of fish sperm physiology and a biological foundation for the further development of spermatozoon motility investigations as well as reproduction technologies. 相似文献
14.
Hollow sperm syndrome during spermatogenesis in the giant tiger shrimp Penaeus monodon (Fabricius 1798) from eastern Australia 下载免费PDF全文
下载免费PDF全文 Roger S‐M Chong Jeff A Cowley Brian D Paterson Greg J Coman David L Mann Stuart J Arnold Howard C Prior Andrew T Wood Mohammed J Amigh 《Aquaculture Research》2015,46(11):2573-2592
During spermatogenesis, giant tiger shrimp (Penaeus monodon) from Queensland, eastern Australia had a high proportion of testicular spermatids that appeared ‘hollow’ because their nuclei were not visible with the haematoxylin and eosin stain. When examined by transmission electron microscopy, the nuclei of hollow spermatids contained highly decondensed chromatin, with large areas missing fibrillar chromatin. Together with hollow spermatids, testicular pale enlarged (PE) spermatids with weakly staining and marginated chromatin were observed. Degenerate‐eosinophilic‐clumped (DEC) spermatids that appeared as aggregated clumps were also present in testes tubules. Among 171 sub‐adult and adult P. monodon examined from several origins, 43% displayed evidence of hollow spermatids in the testes, 33% displayed PE spermatids and 15% displayed DEC spermatids. These abnormal sperm were also found at lower prevalence in the vas deferens and spermatophore. We propose ‘Hollow Sperm Syndrome (HSS)’ to describe this abnormal sperm condition as these morphological aberrations have yet to be described in penaeid shrimp. No specific cause of HSS was confirmed by examining either tank or pond cultured shrimp exposed to various stocking densities, temperatures, salinities, dietary and seasonal factors. Compared with wild broodstock, HSS occurred at higher prevalence and severity among sub‐adults originating from farms, research ponds and tanks. Further studies are required to establish what physiological, hormonal or metabolic processes may cause HSS and whether it compromises the fertility of male P. monodon. 相似文献
15.
Spermiation time affect the milt quality indices of the Russian sturgeon,Acipenser gueldenstaedtii,Brandt & Ratzeburg, 1833 下载免费PDF全文
下载免费PDF全文 Mostafa Halimi Asma Mohammadi Rouhallah Norousta Hossein Khara Mohammad Reza Karimi 《Aquaculture Research》2015,46(10):2426-2430
In this study, the effects of spermiation time are investigated on milt quality of Russian sturgeon over the course of the spawning season. The milt samples were collected from three broodstock batches at three time points including: the beginning, middle and at the end of the spawning season. According to the results, the milt quality parameters including pH, sperm density, spermatocrit, duration of sperm motility and percentage of sperm motility were significantly low in the beginning and end of season than middle of season. The values of milt quality parameters in the middle of season were as follows: (motility percentage: 69.6 ± 3.5, motility duration: 460.3 ± 37.2 s, sperm density: 8.7 ± 0.4 × 109, milt volume: 86.3 ± 8.1 and milt pH: 8.3 ± 0.15). Significant positive correlations were also found between milt pH and sperm motility as well as between sperm density and spermatocrit. In conclusion, our study showed that the middle of season is the best time for collection of milt with appropriate quality in Russian sturgeon. Selection of milt with good quality is necessary aim to cryopreservation of spermatozoa in endangered fish species including Russian sturgeon. 相似文献
16.
Yusuf Bozkurt Fatih Öğretmen Uğur Erçin Ümmügülsüm Yιldιz 《Aquaculture Research》2008,39(15):1666-1672
In this research, the mineral and organic composition of the seminal plasma, physical spermatological parameters and their physiological relationships were investigated in grass carp (Ctenopharyngodon idella). The seminal plasma contained 98.14±5.23 mM L?1 (Na+), 380.85±25.95 mM L?1 (K+), 30.25±4.96 mg dL?1 (Ca2+), 19.16±1.70 mEq L?1 (Mg2+), 1.36±0.11 mg dL?1 glucose, 0.37±0.08 g dL?1 total protein, 12.02±1.18 mg dL?1 cholesterol, 14.85±1.50 mg dL?1 triglyceride and 43.5±9.56 mg dL?1 urea. The following spermatological parameters were found: sperm volume 14.44±1.16 mL, sperm motility 80.60±1.55%, movement duration 67.68±4.32 s, density 15.43±0.72 × 109 mL?1, total density 337.43+45.86 × 109 and pH 7.24±0.17. The Na+ and Ca2+ ions correlated negatively with spermatozoa motility (r=?0.453, P>0.05 and r=?0.192, P>0.05) respectively. The K+ ion correlated positively with spermatozoa motility (r=0.545, P>0.05). But a statistically significant correlation was not observed between sperm motility and seminal plasma parameters. The following correlations were observed between mineral and organic components. The Mg2+ was positively correlated with glucose and cholesterol (r=0.692, P<0.05 and r=0.680, P<0.05) respectively. A highly significant positive relationship was also found between Mg2+ and total protein (r=0.837, P<0.01). On the other hand, a significantly negative relationship was found between Ca2+ and triglyceride (r=?0.639, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of grass carp sperm. 相似文献
17.
Shimelly Rocha Danilo P Streit Jr Lis S Marques Antonio S Varela Jr Carine D Corcini Márcio A Hoshiba 《Aquaculture Research》2018,49(2):963-968
Heavy metals are highly toxic elements that are present in the environment, especially in water. Mercury chloride (HgCl2) stands out among these compounds because of its strong ability to induce damage to any tissue with which it comes into contact. The gametes of spawning aquatic animals, such as fish, are susceptible to such damage. Thus, our objective was to evaluate the toxic potential of HgCl2 in the capacitation and activation of Rhamdia quelen sperm. Semen was collected from seven males and activated in 58 mM sodium chloride (NaCl) containing 0 (control), 4?10, 7?10, 7?9, and 7?8 M HgCl2. The evaluated variables included motility, vigor, motility time, morphology, membrane integrity, membrane fluidity, mitochondrial functionality, production of reactive oxygen species (ROS), and DNA fragmentation. All evaluated HgCl2 concentrations increased primary pathologies and reduced motility, vigor and motility time. Damage to membrane integrity and fluidity began occurring at a concentration of 7?10 M HgCl2. These results indicate that HgCl2 has a toxic effect on different sites of fish spermatozoa and that sperm motility decreases after exposure to HgCl2, impairing sperm capacitation and activation. 相似文献
18.
Pablo E Cruz-Casallas Dora A Lombo-Rodríguez & Yohana María Velasco-Santamaría 《Aquaculture Research》2005,36(7):682-686
In order to develop artificial reproduction in freshwater fish for potential species to be developed in South American aquaculture, milt quality and sperm morphology were studied in yamú (Brycon siebenthalae) under captive conditions during the natural middle spermiation period. The volume of milt collected for each male was 1.8±1.2 mL and the sperm concentration was 13.9±4.0 × 109 spermatozoa mL?1. Spermatocrit (41.5±10.8%) was positively associated (r2=0.30) with sperm density calculated using a corpuscle counting chamber. Sperm motility was 88±9% and the average duration of forward motility was 41±7 s. Fertilization rate was 84±8% and there was no association between this trait and sperm motility (r2=0.009) or with sperm density (r2=0.073). These results suggest that captive B. siebenthalae broodstock can be reproduced successfully. 相似文献
19.
Relationships between body size and secondary sexual characters,and sperm characters in male Dolly Varden char (Salvelinus malma) 下载免费PDF全文
下载免费PDF全文 Toshiaki Yamamoto Noritaka Hirohashi Eiji Fujiwara Tatsuya Suzuki Hatsuna Maruta Hirotake Omiya Shigeru Kitanishi 《Ecology of Freshwater Fish》2017,26(3):397-402
In Salmonidae, subordinate males are exposed to higher risks of sperm competition than dominant males and thus are expected to improve the sperm characteristics (sperm concentrations, sperm velocity and sperm longevity). In this study, we investigated the relationships between body size and secondary sexual characters (breeding colour, hump height and snout length), and sperm characteristics of one‐year‐old (newly matured) Dolly Varden char. Small males displayed higher sperm concentrations than large males. Moreover, males with dull breeding colours, but not with lesser snout length and hump height, displayed an increased sperm velocity compared to males with bright colours, suggesting a trade‐off between sperm quantity and the investment in breeding colour. In addition, sperm longevity decreased as sperm swimming velocity increased. These findings indicate that small males with dull breeding colours improve the quantity and quality of their sperm to a great extent to enhance their chances of reproductive success. 相似文献
20.
计算机辅助对几种鲟鱼冻精激活液的比较 总被引：3，自引：1，他引：3
首次在我国采用鱼类彩色精子图文分析系统(FSQAS-2000)对中华鲟、史氏鲟、西伯利亚鲟和匙吻鲟经超低温冷冻保存后的精子,在不同激活液条件下,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)、平均移动角度(MAD)、以及冷冻前后精子活率的变化等多项参数进行了统计学比较研究。研究分析了不同渗透压、pH以及含Mg~(2 )的激活液对这些参数的影响。研究结果表明:几种鲟鱼激活液的渗透压均较低,其中西伯利亚鲟最低,为10 mOsm·kg~(-1);中华鲟和史氏鲟为20 mOsm·kg~(-1);匙吻鲟最高,为30 mOsm·kg~(-1)。不同的渗透压对冷冻精子的VCL、VSL、VAP、以及精子活率均产生显著(P≤0.05)或极显著(P≤0.01)影响。激活液的pH除了对冷冻精子的上述参数产生影响外,最显著的是对精子的运动轨迹,即平均移动角度(MAD°·s~(-1))产生影响(P≤0.01)。pH越低,精子的MAD越小。几种鲟鱼的最适pH分别为西伯利亚鲟pH 7.5,中华鲟、史氏鲟和匙吻鲟pH 8.5。当4种鲟鱼的冷冻精子激活液中加入5 mmol·L~(-1)MgCl_2时,冷冻精子的3种运动速度显著提高(P≤0.01)。综合分析后认为:西伯利亚鲟的冷冻精子激活液应为10 mmol·L~(-1)Tris-HCl 5 mmol·L~(-1)MgCl_2,pH 7.5;中华鲟和史氏鲟的冷冻精子激活液为20 mmol·L~(-1)Tris-HCl 5 mmol·L~(-1)MgCl_2,pH 8.5;匙吻鲟的冷冻精子激活液为30 mmol·L~(-1)Tris-HCl 5 mmol·L~(-1) MgCl_2,pH 8.5。用此激活液对冷冻前后4种鲟鱼精子活力的相关性进行线性回归分析发现,西伯利亚鲟的R~2=0.8296(P<0.01);中华鲟的R~2=0.9860(P<0.01);史氏鲟的R~2= 0.9622(P<0.01);匙吻鲟R~2=0.9477(P<0.01)。说明冷冻前后4种鲟鱼精子的平均活率存在着极显著的线性正相关性。 相似文献