Jorge-Neto Pedro Nacib, Luczinski Thiago Cavalheri, de Araújo Gediendson Ribeiro, Requena Letícia Alecho, de Jesus Rogério Silva, Souza Larissa Schneider Brandão, Zanella Ricardo, da Costa E Silva Eliane Vianna, de Deco-Souza Thyara, Pizzutto Cristiane Schilbach
Instituto Reprocon, Campo Grande, MS, Brasil.
Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brasil.
Anim Reprod. 2023 Apr 3;20(1):e20230009. doi: 10.1590/1984-3143-AR2023-0009. eCollection 2023.
The cryopreservation of jaguar semen must be improved to produce high-quality biobanking doses. Until now, the rare studies of semen freezing in the species have only evaluated glycerol, always with a significant reduction in sperm quality in thawed semen. The purpose of this study was to assess the efficacy of three cryoprotectants, dimethylsulfoxide (DMSO), glycerol (GLY), and methanol (MET), in the cryopreservation of jaguar semen in an LDL-based extender, as well as the effect of thawing temperature on dosage quality. Five mature males with a history of reproduction were used. On the males, an infrared thermal image (IRT) was captured, the spicules and testes were analyzed, and the CASA system was used to evaluate the quality of fresh and thawed sperm. The superficial IRT was 4.6 ± 1.2 °C cooler than the anal sphincter, and the semen measured between 27.3 and 28.7 °C shortly after exiting the urethra. The total motility of fresh sperm was 55.3 ± 22.6%, and progressive motility was 36.3 ± 18%. The total motility of thawed sperm was 5.28 ± 2.51%, 4.49 ± %2.49, and 0.51 ± 0.62% for DMSO, GLY, and MET, respectively. DMSO and GLY performed better than MET, and there was no difference in thawing temperature (37°C 30 s vs. 50°C 12 s). All animals exhibit a considerable level of morphological changes in sperm. Low amounts of total and progressive motility were found in the thawed sperm. Males with a high level of sperm morphological changes were found to be fertile, but the lone male with normospermia was infertile. Thus, we contest the applicability of the commonly used morphological classification for bovines to felid species.
美洲豹精液的冷冻保存方法必须加以改进,以制备高质量的生物样本库剂量。到目前为止,该物种中关于精液冷冻的少数研究仅评估了甘油,解冻后的精液中精子质量总是显著下降。本研究的目的是评估三种冷冻保护剂,即二甲基亚砜(DMSO)、甘油(GLY)和甲醇(MET),在基于低密度脂蛋白的稀释剂中对美洲豹精液冷冻保存的效果,以及解冻温度对样本质量的影响。使用了五只具有繁殖史的成年雄性美洲豹。对这些雄性美洲豹,采集了红外热图像(IRT),分析了阴茎骨和睾丸,并使用计算机辅助精子分析(CASA)系统评估新鲜精子和解冻后精子的质量。体表IRT比肛门括约肌温度低4.6±1.2°C,精液在离开尿道后不久温度在27.3至28.7°C之间。新鲜精子的总活力为55.3±22.6%,前向运动率为36.3±18%。解冻后精子的总活力,DMSO组为5.28±2.51%,GLY组为4.49±2.49%,MET组为0.51±0.62%。DMSO和GLY的效果优于MET,解冻温度(37°C 30秒与50°C 12秒)之间没有差异。所有动物的精子均表现出相当程度的形态变化。解冻后的精子中总活力和前向运动率较低。发现精子形态变化程度高的雄性具有生育能力,但唯一精子正常的雄性不育。因此,我们质疑常用于牛的形态学分类对猫科动物的适用性。