Cryopreservation of the endangered mahseer (Tor khudree) spermatozoa:: I.: Effect of extender composition, cryoprotectants, dilution ratio, and storage period on post-thaw viability
Cryopreservation of the endangered mahseer (Tor khudree) spermatozoa:: I.: Effect of extender composition, cryoprotectants, dilution ratio, and storage period on post-thaw viability
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DOI：
10.1016/j.cryobiol.2004.05.007
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发表时间：
2004-10-01
期刊：
影响因子：
2.7
通讯作者：
Hegde, SN
中科院分区：
文献类型：
--
作者：
Basavaraja, N;Hegde, SN
提出了几种就地和移地保护策略，以恢复受威胁物种托尔库德雷（Sykes）的种群。精子冷冻保存对于保护濒危物种种群，确保可持续生产至关重要。在不同的保藏剂中，改良鱼林杰（E-1）保藏效果最好。扩展器E-2是次佳选择。以鸡蛋黄和奶粉为原料的膨化剂不适合用于库德氏精子的冷冻保存。在冷冻保护剂中，二甲亚砜在冷冻和解冻过程中对精子的保护作用最大。丙二醇和甲醇的效果较差。在四种稀释剂中，1:10、1:15和1:20的精子运动率优于1:5。在前两种稀释比例下，冷冻前95%以上的运动率分别降至80- 81%和43-67%，冷冻后10天和70天。在所有稀释比下，随着贮藏时间的增加，运动持续时间差异不大。运动率一般随冷冻贮藏时间的增加而降低。精子在25℃解冻保存不同时间后，随保存时间的延长，精子活力率和存活期逐渐降低；解冻后保存40 min的精子仍保持55%的活力，运动时间长达77 s；这些数值进一步下降，导致储存后70分钟完全停止运动。本文讨论了延长剂-冷冻保护剂混合物、稀释剂和储存期在制定库德氏T.精子冷冻保存方案中的重要性。(C) 2004爱思唯尔公司版权所有。
Several in situ and ex situ conservation strategies have been suggested for the revival of stocks of Tor khudree (Sykes), a threatened species. Cryopreservation of spermatozoa is crucial for the conservation of stocks of endangered species so that sustainable production can be ensured. Among the different extenders, modified fish Ringer (E-1) was found to be the best for cryopreservation of T. khudree spermatozoa. Extender E-2 appeared the next best. Extenders based on chicken egg yolk and milk powder were found to be unsuitable for the cryopreservation of T. khudree spermatozoa. Among the cryoprotectants, dimethyl sulfoxide provided maximum protection to spermatozoa during freezing and thawing. Propylene glycol and methanol were found to be less effective. Of the four spermatozoa dilutions, 1:10, 1:15, and 1:20 showed better motility rates than 1:5. At the former dilution ratios, the motility rates which were more than 95% prior to freezing were reduced to 80-81 and 43-67%, 10 and 70 days after cryopreservation, respectively. The motility duration did not differ much with increasing storage period at all the dilution ratios. Motility rates generally decreased with an increase in frozen storage. When spermatozoa were thawed and stored at 25 degreesC for varying periods, motility percentage, and duration decreased gradually as the storage period increased; spermatozoa stored up to 40 min after thawing retained 55% motility and were motile up to 77 s; these values declined further leading to the complete cessation of motility 70 min after storage. The importance of extender-cryoprotectant mixture, milt dilution, and storage period in developing a protocol for T. khudree spermatozoa cryopreservation is discussed. (C) 2004 Elsevier Inc. All rights reserved.