Department of Animal Science, Faculty of Animal Science, Universitas Sebelas Maret, Surakarta, Indonesia.
Department of Animal Production, Faculty of Animal Husbandry, Universitas Padjajaran, Bandung, Indonesia.
Open Vet J. 2024 Feb;14(2):707-715. doi: 10.5455/OVJ.2024.v14.i2.11. Epub 2024 Feb 29.
Epididymal sperm preservation is a simple conservation approach that can help prevent the loss of high genetic quality of farm animals. The chance of loss increases, especially during disease outbreaks or other interruptions to normal reproduction function.
This study looked into the ability of preserved ram epididymal sperm to fertilize oocytes. Due to motility becoming an issue following sperm storage for fertilization, the sperm microinjection known as intracytoplasmic sperm injection approach was employed.
The study was divided into two parts. First, involved the preservation of epididymal sperm at 5°C for 12 days. During preservation, sperm quality parameters namely motility, viability, intact membrane, acrosome, and Deoxyribonucleic acid (DNA) are evaluated every three days. For the fertility test in the second experiment, matured oocytes were injected with immotile sperm discovered in the last days of preservation. The presence of pronucleus development following culture is used as an indicator of sperm's ability to activate and fertilize oocytes.
All sperm quality parameters significantly ( < 0.05) declined during preservation time. On day 12, motility was discovered to be 0%, but viable sperm, sperm with intact membrane, acrosome, and DNA remained at 41.86% ± 9.30%, 31.18% ± 5.15%, 21.88% ± 1.93%, and 33.35% ± 8.74%, respectively. On the fertility test, we inject immotile sperm from day 12 of preservation, which has the lowest motility found, into matured oocytes. Those sperms are able to activate (52.05% ± 7.15%) and fertilize (31.37% ± 1.75%) the injected oocytes, but their fertilizing ability is significantly lower ( < 0.05) when compared to the sperm derived from the ejaculate.
In this study, simple preservation of epididymal sperm reduces all sperm quality criteria, particularly motility. Using the microinjection approach preserved sperm which had no motility, still demonstrated its ability to activate and fertilize the oocytes. According to that, this study provides potential approaches and tools for using genetically superior animals that have lost their ability to execute regular fertilization, and also prolong reproduction function.
附睾精子保存是一种简单的保存方法，可以帮助防止农场动物高遗传质量的损失。在疾病爆发或其他正常繁殖功能中断期间，这种损失的机会增加。
本研究探讨了保存公羊附睾精子的受精能力。由于精子储存后运动能力成为问题，因此采用了精子微注射技术，即胞浆内精子注射法。
该研究分为两部分。首先，将附睾精子在 5°C 下保存 12 天。在保存过程中，每隔三天评估精子质量参数，包括运动能力、活力、完整膜、顶体和脱氧核糖核酸（DNA）。在第二个实验的生育力测试中，将在保存最后几天发现的不活动精子注入成熟的卵母细胞中。培养后出现原核发育被用作精子激活和受精卵母细胞能力的指标。
所有精子质量参数在保存期间均显著（<0.05）下降。第 12 天，发现运动能力为 0%，但存活精子、完整膜精子、顶体精子和 DNA 仍分别为 41.86%±9.30%、31.18%±5.15%、21.88%±1.93%和 33.35%±8.74%。在生育力测试中，我们将在保存的第 12 天发现的运动能力最低的不活动精子注入成熟的卵母细胞中。这些精子能够激活（52.05%±7.15%）和受精（31.37%±1.75%）注入的卵母细胞，但与来自精液的精子相比，其受精能力显著降低（<0.05）。
在这项研究中，简单的附睾精子保存降低了所有精子质量标准，特别是运动能力。使用微注射方法保存的没有运动能力的精子仍然表现出激活和受精卵母细胞的能力。根据这一点，本研究为使用失去正常受精能力的遗传优良动物提供了潜在的方法和工具，并延长了繁殖功能。
