ABSTRACT
BACKGROUND: Sperm cryopreservation of cockerels is a major challenge, and so far there is no adequate information to enable commercial use of frozen semen. OBJECTIVE: To test the toxicity of dimethylacetamide (DMA). MATERIALS AND METHODS: DMA was added at 3%, 6%, 9% and 12% to the freezing diluent, and maintained for equilibration with the semen sample for 1 min, 3 min, 5 min, 7 min and 9 min prior to freezing. Thawed semen was evaluated for kinetic characteristics by computer-assisted semen analysis (CASA) and for structural and functional properties by flow cytometry (plasma membrane rupture, mitochondrial functionality and plasma membrane functionality). RESULTS AND CONCLUSION: The addition of 6% DMA for 3-min equilibration resulted in the highest total and progressive motility, 42.0% and 36.9%, respectively. The point of intersection between a good protection and low plasma membrane rupture was obtained with the addition of 6% of DMA for 3-min equilibration with the rooster semen.
Subject(s)
Acetamides/pharmacology , Chickens , Cryopreservation/veterinary , Semen Preservation/veterinary , Animals , Cryoprotective Agents/pharmacology , Freezing , Male , Semen , Semen Analysis , Sperm Motility , SpermatozoaABSTRACT
ããBACKGROUND: Supplementation of sperm diluents to reduce the damage caused by the freeze-thaw cycle is broadly used in equine semen cryopreservation. OBJECTIVE: The present study aimed at determining the most appropriate quercetin supplementation in equine freezing extender. MATERIALS AND METHODS: Quercetin at four different concentrations (0.25, 0.5, 0.75 or 1 mM) was added in the sperm freezing diluent before the freeze-thaw cycle. The spermatozoa population was analyzed by flow cytometry and a statistical analysis was conducted to detect significant differences between control and treated samples. RESULTS: The statistical analysis did not reveal any significant modification of seminal parameters. CONCLUSION: Within the concentrations tested, quercetin supplementation in equine freezing extender did not affect progressive motility, mitochondrial functionality, acrosome reaction, membrane integrity or DNA fragmentation index in post-thaw equine semen.
