The objective of this study was to compare the BotuCrio ® extender with the Merk-egg yolk and the INRA 82 modified by the inclusion of acetamide, methyl cellulose and trehalose in substitution of glycerol for freezing equine semen. The semen was diluted after centrifugation to obtain 100 x 10 6 of sperm/ml in: BotuCrio ® (control); Merk-egg yolk or INRA 82 modified (Experiment 1). The extended semen was packaged in 0.5 ml straws, cooled and frozen in a freezing machine. The control extender was superior in preserving the motility, VCL, VSL, VAP, LIN, STR and the BCF when compared to the Merk-egg yolk and INRA 82 modified (P < 0.05). The BotuCrio® preserved more effectively the equine sperm viability characteristics evaluated in Experiment 1 and was used as a control extender in Experiment 2 to test the effectiveness of using LDL in replacement of egg yolk. BotuCrio ® was superior to preserve progressive motility, VCL, VSL, VAP, LIN, STR and the percentage of functional integrity of sperm membranes compared to BotuCrio LDL (P < 0.05). However, both extenders preserved similarly the total motility, ALH, BCF and the structural integrity of the membranes (P > 0.05). The fertility rate after AI with frozen semen in BotuCrio LDL was 37.5%.
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