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Cryopreservation of Trans-Andean shovelnose catfish (Sorubim cuspicaudus) semen using dimethylacetamide

机译:使用二甲基乙酰胺冷冻保存反安第斯扁鼻cat鱼(Sorubim cuspicaudus)精液

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Cryoprotectant solutions of dimethylacetamide (DMA) were used at three inclusion levels (8, 10, and 12%) to evaluate cryopreservation of Trans-Andean shovelnose catfish semen. The solutions also included 6% glucose and 5% skimmed milk powder. Osmolarity of the solutions was measured with an osmometer (Precision System, Osmomette III, USA). Semen was diluted at a 1:4 ratio (semen:solution) and frozen in nitrogen vapors using a dry shipper (MVE 4/2V, USA). Concentration, mobility, speed, and progressivity of cryopreserved and fresh semen (control) were assessed with the SCA® Sperm Class Analyzer (Microptic, Spain) computer-assisted program. Fertility and hatching were evaluated fertilizing one gram of oocytes with cryopreserved and fresh semen, which was then kept in 2L cylinder-conical incubators. The osmolarity of cryoprotective solutions was 1233.3 ± 23.1 mOsm/Kg (8%DMA), 1530 ± 0.0 mOsm/Kg (10% DMA), and 1627.7 ± 5.8 mOsm/Kg (12% DMA). Fresh semen showed better total mobility (99.6 ± 0.4%), percentage of rapid sperm (34.7 ± 12.2%), and progressivity (48.2 ± 12.8%), compared with cryopreserved semen (p0.05). Cryopreserved semen with 8% DMA had the highest total mobility (55.4 ± 13.6%) (p0.05) as well as the highest percentage of rapid sperm (1.5-2.5%), total progressivity (1.9 to 10.1%), and curvilinear velocity (29,5-35,9 μm/s) without significant difference with the evaluated DMA percentages (p 0.05). Fertilization with fresh semen (19.5 ± 4.4%) and 8% DMA cryopreserved semen (17.8 ± 8.3%) were not different (p0.05). A cryoprotective solution composed of 8% DMA, 6% glucose, and 5% skimmed milk powder is a viable alternative to cryopreserve Trans-Andean shovelnose catfish semen.
机译:使用二甲基乙酰胺(DMA)的冷冻保护剂溶液以三种包含水平(8%,10%和12%)来评估Trans-Andean shovelnose cry鱼精液的冷冻保存。该解决方案还包括6%的葡萄糖和5%的脱脂奶粉。用渗透压计(Precision System,Osmomette III,美国)测量溶液的渗透压。精液以1:4的比例(精液:溶液)稀释,并使用干托运人(MVE 4 / 2V,美国)在氮气中冷冻。用SCA®Sperm Class Analyzer(西班牙Microptic)计算机辅助程序评估冷冻保存和新鲜精液(对照)的浓度,迁移率,速度和累进性。用冷冻保存的新鲜精液对1克卵母细胞进行受精,从而评估其受精能力和孵化率,然后将其保存在2L圆柱锥形培养箱中。冷冻保护溶液的渗透压为1233.3±23.1 mOsm / Kg(8%DMA),1530±0.0 mOsm / Kg(10%DMA)和1627.7±5.8 mOsm / Kg(12%DMA)。与冷冻保存的精液相比,新鲜精液显示出更好的总迁移率(99.6±0.4%),快速精子百分比(34.7±12.2%)和渐进性(48.2±12.8%)。具有8%DMA的冷冻精液具有最高的总迁移率(55.4±13.6%)(p <0.05),并且具有最高的快速精子百分比(1.5-2.5%),总进行性(1.9至10.1%)和曲线速度(29,5-35,9μm/ s)与评估的DMA百分比没有显着差异(p> 0.05)。新鲜精液(19.5±4.4%)和8%DMA冷冻精液(17.8±8.3%)的受精率没有差异(p> 0.05)。由8%的DMA,6%的葡萄糖和5%的脱脂奶粉组成的冷冻保护溶液可以替代冷冻储备的Trans-Andean shovelnose fish鱼精液。

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