Bernáth G, Bokor Z, Żarski D, Várkonyi L, Hegyi Á, Staszny Á, Urbányi B, Radóczi Ifj J, Horváth Á
Department of Aquaculture, Szent István University, Páter Károly 1., H-2100 Gödöllő, Hungary.
Department of Aquaculture, Szent István University, Páter Károly 1., H-2100 Gödöllő, Hungary.
Anim Reprod Sci. 2016 Jul;170:170-7. doi: 10.1016/j.anireprosci.2016.05.005. Epub 2016 May 13.
The quality and fertilizing capacity of perch (Perca fluviatilis) sperm collected outside of the spawning season (off-season) and cryopreserved at a commercial scale, were tested. Basic parameters (equilibration time, dilution ratio, sperm concentration, post-thaw motility duration) which can have a significant effect on cryopreservation success were systematically investigated for effects on sperm quality using computer assisted sperm analysis (CASA). No significant decrease in progressive motility (pMOT) and straightness (STR) of fresh-diluted sperm was recorded among groups equilibrated for 0, 30 or 60min in an extender with cryoprotectants. Curvilinear velocity (VCL) was reduced significantly after 30min (30min: 146±15μm/s, 60min: 124±18μm/s) of equilibration compared to the control (174±9μm/s). After thawing, no decrease in pMOT or VCL was observed at different equilibration times in any of the analyzed groups. No correlation was observed among progressive motility, dilution ratios (p=0.7) and cell concentrations (p=0.1). The use of different activating solutions resulted in similar pMOT and VCL in the first 120s post-thaw. Nevertheless, post-thaw sperm motility was reduced after 30s using all activators. Motility parameters with low variation were recorded after thawing of 57 straws (pMOT: 37±7%, VCL: 92±10μm/s, STR: 89±3%). Ten randomly selected straws from commercial-scale cryopreservation resulted in a high fertilization rate (cryopreserved sperm: 72±14%, fresh control: 94±2%). An optimized commercial-scale cryopreservation protocol was successfully developed for Eurasian perch. The applicability of the off-season collected perch sperm for cryopreservation and fertilization was demonstrated.
对在产卵季节之外（非繁殖季）采集并进行商业规模冷冻保存的河鲈（Perca fluviatilis）精子的质量和受精能力进行了测试。使用计算机辅助精子分析（CASA）系统研究了可能对冷冻保存成功有显著影响的基本参数（平衡时间、稀释比例、精子浓度、解冻后活力持续时间）对精子质量的影响。在含有冷冻保护剂的稀释液中平衡0、30或60分钟的各组中，新鲜稀释精子的直线运动速度（pMOT）和直线性（STR）均未出现显著下降。与对照组（174±9μm/s）相比，平衡30分钟（30分钟：146±15μm/s，60分钟：124±18μm/s）后曲线速度（VCL）显著降低。解冻后，在任何分析组中，不同平衡时间下均未观察到pMOT或VCL下降。在直线运动速度、稀释比例（p = 0.7）和细胞浓度（p = 0.1）之间未观察到相关性。使用不同的激活溶液在解冻后的前120秒内产生了相似的pMOT和VCL。然而，使用所有激活剂在30秒后解冻后精子活力均降低。解冻57支细管后记录到活力参数变化较小（pMOT：37±7%，VCL：92±10μm/s，STR：89±3%）。从商业规模冷冻保存中随机选择的10支细管产生了较高的受精率（冷冻保存精子：72±14%，新鲜对照组：94±2%）。成功开发了一种针对欧亚河鲈的优化商业规模冷冻保存方案。证明了非繁殖季采集的河鲈精子用于冷冻保存和受精的适用性。
