Master Program in Biology, Faculty of Matemathics and Natural Sciences, Syiah Kuala University, Banda Aceh, 23111, Indonesia.
Department of Aquaculture, Faculty of Marine and Fisheries, Syiah Kuala University, Banda Aceh, 23111 Indonesia.
Cryo Letters. 2023 May-Jun;44(3):169-177.
Goldfish Carassius auratus is a popular ornamental fish extensively cultured worldwide. Sperm cryopreservation is a common fish breeding method that ensures sperm availability around the year. Studies on cryopreservation of goldfish sperm, especially on the suitability of cryoprotectant types and pre-freezing time, are scarcely available.
To determine the most suitable type of cryoprotectant and pre-freezing for the successful cryopreservation of goldfish sperm.
A completely randomized design with two factors was utilized in this study. The first factor is the type of cryoprotectants, which included methanol, ethanol, ethylene glycol, glycerol, and DMSO. The second is pre-freezing times of 10, 20, 30, and 40 min at each of the pre-freezing temperatures of 4 degree C, -10 degree C, and -79 degree C, meaning that the total times for the ramping down of temperature were 30, 60, 90 and 120 min, respectively. The Ringer solution and 10% egg yolk were used as extender and extracellular cryoprotectant. The sperm was stored at -179 degree C for 7 days.
The ANOVA test showed that cryoprotectants and pre-freezing significantly affected the motility, viability, and fertility of goldfish sperm after freezing in liquid nitrogen for 7 days (P<0.05). Furthermore, 10% DMSO combined with 15% egg yolk with an pre-freezing time of 20 min can maintain sperm motility, viability, and fertility higher than other treatments, by 79%, 80%, and 33%, respectively. The agarose gel electrophoresis showed no DNA fragmentation in all samples, including fresh sperm.
We conclude that 10% DMSO combined with 15% egg yolk and 20 min pre-freezing is the best treatment for goldfish sperm cryopreservation. DOI: 10.54680/fr23310110412.
金鱼(Carassius auratus)是一种广泛养殖的受欢迎的观赏鱼。精子冷冻保存是一种常见的鱼类繁殖方法,可确保全年都有精子供应。关于金鱼精子冷冻保存的研究,特别是关于冷冻保护剂类型和预冷冻时间的适用性的研究很少。
确定最适合金鱼精子冷冻保存的冷冻保护剂类型和预冷冻时间。
本研究采用完全随机设计,有两个因素。第一个因素是冷冻保护剂的类型,包括甲醇、乙醇、乙二醇、甘油和 DMSO。第二个因素是在 4°C、-10°C 和-79°C 的预冷冻温度下分别预冷冻 10、20、30 和 40 分钟,这意味着温度下降的总时间分别为 30、60、90 和 120 分钟。林格氏溶液和 10%卵黄用作扩展剂和细胞外冷冻保护剂。精子在-179°C 下储存 7 天。
方差分析表明,冷冻保护剂和预冷冻时间对金鱼精子在液氮中冷冻 7 天后的活力、存活率和受精率有显著影响(P<0.05)。此外,10%DMSO 与 15%卵黄结合,预冷冻时间为 20 分钟,可以保持精子活力、存活率和受精率分别高于其他处理方式,分别为 79%、80%和 33%。琼脂糖凝胶电泳显示所有样本(包括新鲜精子)均无 DNA 片段化。
我们得出结论,10%DMSO 与 15%卵黄和 20 分钟预冷冻是金鱼精子冷冻保存的最佳处理方法。DOI:10.54680/fr23310110412。