L-proline: an effective agent for donkey frozen and post-thawed semen storage

Abstract The aim of this study was to evaluate the effects of L-proline on the extender quality of frozen and post-thawed jackass semen. Jackass (n = 6) semen samples were collected and cryopreserved in gradient concentrations (0–80 mM) of L-proline in extenders; post-thawed semen samples were cultured in L-proline medium for 10 hours at 37°C. For cryopreservation experiment I, the motile parameters, mitochondrial membrane potential (MMP), and plasma membrane, acrosome, and chromatin structure integrities of post-thawed semen were assessed. For culture experiment II, additional ROS contents were analyzed after incubation. For the fertility trial, jennies (n = 135) were divided into group I (30 mM L-proline in cryopreservation extender), group II (40 mM L-proline in culture medium), and the control. Pregnancy was diagnosed using an ultrasound scanner 30 days after ovulation. The results of experiment I showed that, motile parameters and acrosome and chromatin structure integrities of groups I and 40 mM were significantly higher than the control (P