Dehydration of llama sperm using different osmolarity media and temperatures for preservation

Abstract

The objective of this study was to evaluate effects of dehydration on sperm DNA with the aim of eventually using this method for preserving llama spermatozoa. Two experiments were conducted: 1) sperm preservation at 5 °C for 60 days in different hyperosmotic solutions (500, 800, 1000 and 1200 mOsmol/l) (n = 6, replications = 2) and 2) sperm preservation at 5 and −20 °C for 60 days in the same hyperosmotic solutions, with supplementary antibiotics (n = 6, replications = 2). Sperm motility, membrane functional integrity, viability and morphology were evaluated at 0 and 48 h of the preservation period (Experiment 1) and at 30 min and 24 h (Experiment 2). Sperm DNA was evaluated at 0 or 30 min (Experiment 1 and 2, respectively) and on days 7, 14, 21, 30 and 60 of the preservation periods. Motility, membrane functional integrity and viability were less when sperm were dehydrated, while sperm cell morphology was not affected. There was a smaller percentage of sperm with condensed chromatin as duration of the preservation period increased when stored in the different hyperosmotic solutions. There was a markedly smaller (P < 0.05) percentage of sperm with intact DNA in all solutions as the duration of preservation increased, with there being greater values for intact DNA at −20 °C than sperm preserved at 5 °C. Llama sperm chromatin condensation was slightly affected by the process of dehydration. There was a markedly smaller percentage of sperm with intact DNA in the dehydrated semen samples.