Effect of Extender and Freezing Rate on Quality Parameters and In Vitro Fertilization Capacity of Alpaca Spermatozoa Recovered from Cauda Epididymis

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.