Effect of different concentrations of Trolox ® in association with docosahexaenoic acid on equine semen freezing.

The aim of this study was to evaluate the association of different concentrations of Trolox ® and the addition of a fixed concentration of DHA in the freezing of semen of Mangalarga Marchador stallions. To that end, 16 ejaculates were frozen in the following extenders: E1) BotuCrio ® (BC; Control); E2) BC + 50 ngml -1 DHA + 30 µM Trolox ® (BCDHA30T); E3) BC + 50 ngml -1 DHA + 40 µM Trolox ® (BCDHA40T); E4) BC + 50 ngml -1 DHA + 50 µM Trolox ® (BCDHA50T). All the tested extenders were similar in preserving different kinematic parameters, cell functional integrity, compacted DNA, and high and intermediate mitochondrial activity (P>0.05). However, sperm cryopreserved in BCDHA40T showed higher velocities than sperm frozen in the control extender (P<0.05). The 30 µM concentration of Trolox ® was worse for sperm motility and the 50 µM concentration of Trolox ® did not adequately preserve the structural integrity of the membranes in an extender containing DHA when compared to the BotuCrio ® (P<0.05) extender. The use of Trolox ® in freezing extenders containing DHA did not maximize the effect of BotuCrio ® , except for in the case of sperm velocity parameters when at a concentration of 40 µM.