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1.
Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocol for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants (ethylene glycol, 1, 2-propanediol,dimethyl sulfoxide and glycerol) were employed at three concentrations (1.0, 1.5 and 2.0 molL-1) respectively and survival rates of larvae were determined after a storage of 1h. The larvae frozen with these four cryoprotectants after 1h storage were cultured, and then survival rates were determined at 24, 72 and 120h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL-1 showed the best protective effect in all experiments (p<0.05). And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1,7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL-1 dimethyl sulfoxide after 1h, 1d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34% ±11.28%, 83.10% ±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods (p>0.05).  相似文献   
2.
Amphioxus, Branchiostoma belcheri, a closest relative of vertebrates, is at a high risk of extinction due to a combination of low effective population size, altered native habitats and environmental pollution, yet little is known about cryopreservation of its gametes. This study deals with the cryopreservation of amphioxus semen. The main findings are that (1) the extender of Yao et al. is the best one among the four extenders examined; (2) the appropriate ratio of semen to extender of Yao et al. plus cryoprotectant is from 1:5 to 1:7; (3) dimethyl sulfoxide (DMSO) and methanol are the better cryoprotectants than glycerol, with DMSO giving the best results; (4) the eggs fertilized with post-thaw spermatozoa are capable of developing to at least hatching stage, and the highest hatching rate is (12.4±3.0)%. This is the first report on freezing and thawing of amphioxus spermatozoa, providing a simple and practical protocol for cryopreservation of amphioxus spermatozoa and laying a foundation for safeguarding this endangered species.  相似文献   
3.
包埋脱水法冷冻保存裙带菜配子体克隆的研究   总被引:6,自引:0,他引:6  
将裙带菜配子体克隆包埋在含3%褐藻酸钙的胶球中,研究了预培养中的蔗糖浓度和培养时间,脱水过程中的脱水速率和胶球含水量以及化冻速度对配子体克隆的存活率和发育能力的影响,结果表明含裙带菜配子体克隆的胶球在0.3 mol/dm3的蔗糖培养基中培养9 h可以获得较高的存活率;胶球最适平均脱水速率为含水量的5%每小时,最适含水量为25%;较快的化冻速度可以显著提高配子体克隆的存活率,在优化条件下存活率可高达66%;经过冷冻保存的裙带菜配子体克隆在适合发育的条件下能够正常形成孢子体.  相似文献   
4.
大黄鱼精液的高效超低温保存   总被引:2,自引:0,他引:2  
采用2 mL的冷存管和程序降温仪高效超低温保存大黄鱼(Pseudosciaena croce)精液。分析比较了5种不同浓度抗冻剂(二甲基亚砜、乙二醇、丙二醇和甘油和甲醇)、3种降温速率(10,20和30℃/min)和2种解冻温度(28℃和43℃)对大黄鱼精液超低温保存效果的影响。实验结果表明,采用Hanks’作为稀释液,10%~20%甘油或10%DMSO作为抗冻剂,20℃/min的降温速率对精液进行超低温保存43℃水浴解冻,冻精激活后获得理想的运动率(76.6%±11.4%~80.0%±12.6%),而且对10%甘油保存的冻精进行人工授精实验获得了与新鲜精液相似的受精率和孵化率。  相似文献   
5.
用包埋脱水法冰冻保存小新月菱形藻   总被引:1,自引:0,他引:1  
采用包埋脱水法研究了小新月菱形藻(Nitzschia closterium minutissima)的超低温保存,探讨了影响存活率的内、外因素:藻细胞年龄、脱水速率、胶球含水量和化冻后的恢复方法等。结果表明,将处于静止初期的藻细胞包埋在含有0.5 mol/L蔗糖的3%褐藻酸钙胶球中,以含水量每小时减少0.9%的平均速率脱水至胶球含水量为40%,化冻后在冻存管中室温下暗恢复12 h的条件下冰冻存活率较高,可达到18.3%左右。与常规的两步法冰冻保存相比,包埋脱水法可大大简化操作程序,是保存小新月菱形藻的一种有潜力的方法。  相似文献   
6.
太平洋牡蛎精液的超低温保存   总被引:1,自引:1,他引:0  
利用解剖法采集太平洋牡蛎精液 ,用自然海水做基础溶液 ,配制不同浓度的 DMSO(二甲亚砜 )作为超低温保护剂 ,在液氮 (L N2 )面上不同高度进行预处理 ,投入 LN2 内保存 ,升温解冻后 ,检测精子成活率。结果表明 ,DMSO处理浓度、预处理温度及时间和升温方式对超低温保存成活率均具有显著影响。其中 ,用 10 % DMSO在 LN2 面上 17cm预处理 6 min,采用 16~ 17℃流水不完全解冻 ,效果最好 ,超低温保存精子成活率可达 70 %。与新鲜精子相比 ,在 L N2 内保存一天的精子 ,授精能力没有明显变化。  相似文献   
7.
锯缘青蟹精子低温冷藏及精子活力的染色法评价   总被引:11,自引:0,他引:11  
管卫兵  王桂忠  李少菁  陈高峰 《台湾海峡》2002,21(4):457-462,T004
采用天然海水配制的台盼蓝染色液和曙约B染色液对低温冷冻的锯缘青蟹精子的活力进行评价研究。结果表明,用这两种染色液染色死活精子呈现出明显不同的特征,进而以无锡离子人工海水(Ca^2 -FSW)为冷冻基础液,分别用10%(V/V)甘油、5%(V/V)二甲亚砜(DMSO)作冷冻剂,进行了4℃液态低温保存的精子实验,对照组不加冷冻剂,以Ca^2 -FSW等量代替,同时采用染色法对保存精子活力进行评价,结果显示台盼蓝不适宜于评价甘油组精子的活力,但却能评价对照组和二甲亚砜组。在24h内,二甲亚砜组精子的存活率达58.7%,而对照组仅为30.2%。可见,染色法可以作为低温冷冻保存的锯缘青蟹精子活力评价的一种有效方法。  相似文献   
8.
The Cawthron Institute Culture Collection of Micro-algae (CICCM) is a unique, nationally and internationally significant collection of 450 strains of approximately 100 micro-algae and 50 cyanobacteria species, both living and cryopreserved. The collection comprises 13 taxonomic classes and underpins research into the ecology and taxonomy of the isolates and the biotoxins they produce and their toxicity. The CICCM expanded significantly in the 1990s after a major harmful algae bloom event in the Hauraki Gulf, New Zealand. Since 2000, it has underpinned development of molecular detection tools and enabled development of new chemical testing methods for biotoxins in seafood. The tropical collection allows research into potential issues for New Zealand as the oceans warm, for example, the risks of ciguatera fish poisoning and palytoxins in seafood. Research results generated from isolates in the cyanobacterial collection have assisted national risk management regarding drinking water and informed the development of the New Zealand guidelines for managing cyanobacteria in recreational fresh waters. The actual living and cryopreserved micro-algae and cyanobacteria collection (without infrastructure) has a current estimated value of NZ$1.6 million.  相似文献   
9.
点带石斑鱼精子超低温冷冻保存研究   总被引:1,自引:0,他引:1  
为建立点带石斑鱼(Epinephelus malabaricus)精子冷冻保存技术,保存点带石斑鱼种质资源,作者通过选用TS-19、TS-2、Hank’s、MPRS、Cortland、0.75%NaCl等作为稀释液及10%~25%二甲亚砜、甘油、甲醇作为抗冻液,对繁殖季节中3个不同时期的点带石斑鱼精液进行超低温冷冻保存研究。研究结果表明:(1)Hank’s溶液为点带石斑鱼精子冷冻保存最适冷冻稀释液;(2)Hank’s溶液与10%~15%DMSO溶液为点带石斑鱼精子保存的最适冷冻保护液组合,可以获得60%以上的冻精成活率,而使用不同浓度的MeOH作为抗冻液时,冻精成活率均为0%;(3)以10%~25%Gly作为抗冻液时,冷冻前未加天然海水时均能激活点带石斑鱼精子正常运动,其效果与天然海水相似,加入天然海水后精子运动效果急剧下降,精子冷冻保存后也存在同样现象;(4)为保证获得更高的冻精成活率,点带石斑鱼精液冷冻保存样品的最佳获取时期是在亲鱼自然繁殖期前进行为宜。  相似文献   
10.
Cryopreservation experiments were conducted on D-stage larvae of the Pacific oyster (Crassostrea gigas) to investigate the effects of two cryoprotectant solutions and three cooling rates on larval development from 1 to 22 days post-fertilisation. Cryoprotectant solutions were made up to final concentrations (after 1:1 dilution with larvae) of 10% ethylene glycol, 1% polyvinylpyrrolidone and either 0.2 or 0.4 M trehalose. Three cooling rates (0.5, 1 and 2 °C min?1 between ?10 and ?35 °C post-holding) were tested in an orthogonal design with the two cryoprotectants. Results indicate that control larvae out-performed all cryopreservation treatments for survival, feeding consumption and shell length parameters. However, larvae exposed to 0.4 M trehalose did considerably better than those exposed to 0.2 M trehalose, regardless of cooling rate conditions. Scanning electron and light microscopy observations were used to assess larval morphology and organogenesis, indicating that treatments with surviving larvae were morphologically and developmentally similar to control larvae.  相似文献   
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