Castelo Thibério de Sousa, Silva Andréia Maria da, Peixoto Gislayne Christianne Xavier, Souza Ana Liza Paz, Campos Lívia Batista, Lima Gabriela Liberalino, Dantas Maiko Roberto Tavares, Souza-Junior João Batista Freire de, Silva Alexandre Rodrigues
Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido, BR 110, Km 47, Costa e Silva, 59625-900, Mossoró, RN, Brazil.
Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido, BR 110, Km 47, Costa e Silva, 59625-900, Mossoró, RN, Brazil.
Cryobiology. 2023 Dec;113:104787. doi: 10.1016/j.cryobiol.2023.104787. Epub 2023 Oct 25.
This study investigated whether the origin of sperm (epididymal vs. ejaculate) affects the cryopreservation efficiency in agouti (Dasyprocta leporina). Five sexually mature agoutis underwent electroejaculation, resulting in obtaining four semen samples. After 15 days, the same animals were euthanized, and through retrograde flushing, sperm samples were obtained from the epididymis tails. In both collection methods, samples were evaluated for sperm parameters (sperm concentration, motility, vigor, membrane integrity, osmotic response, and morphology). Then, samples were diluted in ACP 109c, added with 20% egg yolk, and a final concentration of 6% glycerol. Finally, the samples were packaged in 0.25 mL straws and frozen in liquid nitrogen. After one week, samples were thawed and evaluated in the same way as fresh samples, with the addition of membrane integrity analysis using fluorescent probes (C-FDA/PI) and computerized analysis (CASA). Immediately after obtaining the sperm, samples obtained directly from the epididymis presented higher values (P ≤ 0.05) than those obtained by electroejaculation concerning the parameters of volume, sperm concentration, and total number of sperm (1,398.25 ± 206.0 x10 and 184.5 ± 78.0 x10 sperm). On the other hand, in the classical evaluation of the other sperm parameters and the computerized analysis (CASA) after thawing, such as total motility, no statistical differences were observed between sperm from both origins (ejaculate: 16.7 ± 8.2% and epididymal: 24.8 ± 12.0%, P > 0.05). This demonstrates the possibility of direct application of the cryopreservation protocol for agouti (D. leporina) sperm obtained via the epididymis or ejaculate.
本研究调查了刺豚鼠（Dasyprocta leporina）精子的来源（附睾精子与射精精子）是否会影响其冷冻保存效率。五只性成熟的刺豚鼠接受电刺激射精，从而获得了四份精液样本。15天后，对相同的动物实施安乐死，并通过逆行冲洗从附睾尾部获取精子样本。在两种采集方法中，均对样本的精子参数（精子浓度、活力、活率、膜完整性、渗透反应和形态）进行评估。然后，将样本在添加20%蛋黄的ACP 109c中进行稀释，并使甘油终浓度达到6%。最后，将样本包装在0.25 mL细管中，并在液氮中冷冻。一周后，将样本解冻，并按照与新鲜样本相同的方式进行评估，同时增加使用荧光探针（C-FDA/PI）的膜完整性分析和计算机辅助分析（CASA）。在获取精子后立即进行评估，就体积、精子浓度和精子总数参数而言，直接从附睾获得的样本（1,398.25 ± 206.0×10和184.5 ± 78.0×10精子）比通过电刺激射精获得的样本具有更高的值（P≤0.05）。另一方面，在解冻后对其他精子参数的经典评估以及计算机辅助分析（CASA）中，例如总活力，两种来源的精子（射精精子：16.7 ± 8.2%和附睾精子：24.8 ± 12.0%，P > 0.05）之间未观察到统计学差异。这表明了对通过附睾或射精获得的刺豚鼠（D. leporina）精子直接应用冷冻保存方案的可能性。
