Várkonyi Levente, Bokor Zoltán, Molnár József, Fodor Ferenc, Szári Zsolt, Ferincz Árpád, Staszny Ádám, Láng Levente Zete, Csorbai Balázs, Urbányi Béla, Bernáth Gergely
Department of Aquaculture, Szent István University, Agárd, Hungary.
Balaton Fish Management Non-Profit Ltd, Siófok, Hungary.
Reprod Domest Anim. 2019 Mar;54(3):639-645. doi: 10.1111/rda.13383. Epub 2018 Dec 15.
In our study, a traditionally used (Grayling, already used in cyprinid species) and a newly tested (Pike) extender was tested to avoid sperm agglutination phenomenon following thawing during carp sperm cryopreservation. A large-scale (elevated volume of sperm) freezing method in a controlled-rate freezer using 5 ml straw and 10 ml cryotube was also systematically established. In all experiments, the sperm cryopreserved in using Grayling extender (except only one sample) showed an agglutination phenomenon (damaged and intact cells adhered to each other) after thawing where Pike extender resulted the regular cell suspension. No significant difference was observed between the two cryopreserved groups (Pike and Grayling extender) in all motility parameters using the 0.5 ml straw and the polystyrene box. Similarly, motility parameters did not show a significant difference in the two frozen groups with the 5 ml straw, also in the polystyrene box. A significantly higher progressive motility (pMOT, Grayling: 54% ± 8%, Pike: 37% ± 5%), straight line velocity (VSL, Grayling: 50 ± 5 µm/s, Pike: 39 ± 4 µm/s) and beat cross frequency (BCF, Grayling: 20 ± 1 Hz, Pike: 17 ± 1 Hz) was observed in the case of the grayling extender by the 5 ml straw cryopreserved in a controlled-rate freezer (CRF) compare to the pike extender. A significantly higher VSL (Grayling: 45 ± 3 µm/s, Pike: 38 ± 4 µm/s) was observed by the grayling extender using the 10 ml cryotube than with the pike extender. Despite the randomly occurring differences in a few parameters, our new controlled freezing method using the newly tested Pike extender, the 5 ml straw or the 10 ml cryotube can be a good solution for the preservation of elevated volume of carp sperm.
在我们的研究中，测试了一种传统使用的（已在鲤科鱼类中使用的茴鱼精液稀释液）和一种新测试的（狗鱼精液稀释液）稀释液，以避免鲤鱼精子冷冻保存解冻后出现精子凝集现象。还系统地建立了一种在程序降温冷冻机中使用5毫升细管和10毫升冻存管的大规模（精子体积增加）冷冻方法。在所有实验中，使用茴鱼精液稀释液冷冻保存的精子（仅一个样本除外）解冻后出现凝集现象（受损和完整细胞相互粘附），而狗鱼精液稀释液则产生正常的细胞悬液。在使用0.5毫升细管和聚苯乙烯盒的所有活力参数方面，两个冷冻保存组（狗鱼精液稀释液和茴鱼精液稀释液）之间未观察到显著差异。同样，在使用5毫升细管且同样置于聚苯乙烯盒中的两个冷冻组中，活力参数也未显示出显著差异。与狗鱼精液稀释液相比，在程序降温冷冻机中使用5毫升细管冷冻保存的情况下，茴鱼精液稀释液的精子表现出显著更高的前进运动率（pMOT，茴鱼：54%±8%，狗鱼：37%±5%）、直线速度（VSL，茴鱼：50±5μm/s，狗鱼：39±4μm/s）和摆动交叉频率（BCF，茴鱼：20±1Hz，狗鱼：17±1Hz）。使用10毫升冻存管时，茴鱼精液稀释液的精子VSL显著更高（茴鱼：45±3μm/s，狗鱼：38±4μm/s）。尽管在一些参数上存在随机出现的差异，但我们使用新测试的狗鱼精液稀释液、5毫升细管或10毫升冻存管的新型程序冷冻方法对于保存大量鲤鱼精子可能是一个很好的解决方案。
