Mamani-Mango Guiulfo, Moina Gonzales Milagros, Ramos Hidalgo Martín, Mendoza Mallma José, Ruiz Béjar Jaime, Rivas Palma Victoria, Mellisho Salas Edwin
1 Department of Animal Production, Faculty of Animal Husbandry, National Agrarian University La Molina, Lima, Peru.
2 Professional School of Veterinary Medicine, Universidad Nacional San Antonio Abad Del Cusco, Cusco, Peru.
Biopreserv Biobank. 2019;17(1):39-45. doi: 10.1089/bio.2018.0021. Epub 2018 Sep 26.
In alpacas, improvement of reproductive efficiency of male camelids is limited by the small testicular size, low spermatozoa production, and low quality of semen. In this study we aim to evaluate the effect of two extenders and two freezing rates on post-thaw quality of sperm recovered from alpaca epididymis with two methods (flushing and mincing), and to evaluate the in vitro fertilization (IVF) capacity of frozen sperm selected with two different selection methods (washing and swim-up). Sperm samples were processed with Tris-egg yolk or Bioxcell extenders and frozen with slow freezing and fast freezing. The oocytes were coincubated with spermatozoa for 72 hours, and cleavage rates were recorded afterward. The results indicated that the recovery method did not influence sperm quality (∼70%). However, total sperm recovery was significantly lower for the flushing method than the mincing method. The sperm quality was influenced by the freezing extender (23.3% vs. 33.2%) and freezing rate (20.9% vs. 35.7%). When comparing different methods of sperm selection for IVF, no differences were observed on cleavage rate except for the fact that the concentration of sperm from swim-up method (20.6%) was significantly lower than the one obtained from the washing method (78.7%). The recovery technique of sperm does not affect sperm quality and the method of fast freezing was shown to be the most effective for cryopreservation of alpaca sperm.
在羊驼中,雄性骆驼科动物繁殖效率的提高受到睾丸体积小、精子产量低和精液质量差的限制。在本研究中,我们旨在评估两种稀释液和两种冷冻速率对通过两种方法(冲洗和切碎)从羊驼附睾回收的精子解冻后质量的影响,并评估用两种不同选择方法(洗涤和上游法)选择的冷冻精子的体外受精(IVF)能力。精子样本用Tris-蛋黄或Bioxcell稀释液处理,并分别采用慢速冷冻和快速冷冻。将卵母细胞与精子共同孵育72小时,之后记录卵裂率。结果表明,回收方法不影响精子质量(约70%)。然而,冲洗法的总精子回收率显著低于切碎法。精子质量受冷冻稀释液(23.3%对33.2%)和冷冻速率(20.9%对35.7%)的影响。比较不同的IVF精子选择方法时,除了上游法获得的精子浓度(20.6%)显著低于洗涤法(78.7%)外,卵裂率未观察到差异。精子的回收技术不影响精子质量,快速冷冻法被证明是羊驼精子冷冻保存最有效的方法。