Ji X S, Zhao Y, Chen S L, Jiang Y L, Wang H, Song J Y, Ding L, Chen H J
Shandong Agricultural University, Daizong Street 61, Taian, 271018, China.
Theriogenology. 2008 Apr 15;69(7):793-7. doi: 10.1016/j.theriogenology.2007.10.022. Epub 2008 Mar 11.
Although sperm from several fish species have been successfully cryopreserved, few studies have been done in small and/or endangered species. The aim of the present work was to develop a method of freezing and refreezing Varicorhinus macrolepis semen in 1.8 mL cryovials. The effect of extenders and cryoprotectants on the motility of post-thaw sperm was examined. The motility of frozen-thawed sperm in extender D-15 was higher than that in MPRS and fish Ringer solution (P<0.05). Dimethyl sulfoxide (DMSO) and glycerol provided greater protection to sperm than methanol during freezing and thawing; the most effective concentration of DMSO and glycerol was 10%. The fertilization rate of frozen-thawed sperm was not significantly different from that of fresh sperm. Furthermore, mean (+/-S.D.) hatching rate did not differ significantly between frozen-thawed (82.7+/-12.4%) and fresh sperm (90.7+/-4.5%). Although frozen-thawed sperm that was immediately refrozen had 0% post-thaw motility, frozen semen that was refrozen after dilution with D-15 (containing DMSO at a ratio of 1:2) had post-thaw motility of 38.3+/-2.9%. Motility was lower for refrozen than for frozen sperm (P<0.05). Furthermore, fertilization and hatching rates of refrozen sperm were 42.9+/-6.7 and 34.1+/-10.5%, respectively, which were lower than that of fresh sperm (P<0.05).
尽管几种鱼类的精子已成功冷冻保存，但针对小型和/或濒危物种的相关研究却很少。本研究的目的是开发一种在1.8毫升冻存管中冷冻和解冻麦氏突吻鱼精液的方法。研究了稀释液和冷冻保护剂对解冻后精子活力的影响。解冻后精子在D-15稀释液中的活力高于改良任氏液（MPRS）和鱼类林格氏液中的活力（P<0.05）。在冷冻和解冻过程中，二甲基亚砜（DMSO）和甘油对精子的保护作用比甲醇更好；DMSO和甘油的最有效浓度均为10%。解冻后精子的受精率与新鲜精子的受精率无显著差异。此外，解冻后精子（82.7±12.4%）和新鲜精子（90.7±4.5%）的平均（±标准差）孵化率也无显著差异。虽然立即重新冷冻的解冻后精子解冻后活力为0%，但用D-15（含1:2比例的DMSO）稀释后重新冷冻的冻存精液解冻后活力为38.3±2.9%。重新冷冻精子的活力低于冷冻精子（P<0.05）。此外，重新冷冻精子的受精率和孵化率分别为42.9±6.7和34.1±10.5%，均低于新鲜精子（P<0.05）。
