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.