title: Effect of staining and freezing media on sortability of stallion spermatozoa and their post-thaw viability after sex-sorting and cryopreservation.
abstract: Sex-sorted, frozen-thawed stallion spermatozoa remain out of reach of commercial horse breeders because of the low efficiency of the sex-sorting process and unacceptable fertility rates after insemination. Two experiments were designed to test the effects of alternative staining and freezing media to improve the viability of sex-sorted frozen-thawed stallion spermatozoa. Experiment 1 compared two freezing media, INRA 82(®)  and a modified lactose-ethylenediaminetetraacetic acid (EDTA), for the cryopreservation of sex-sorted stallion spermatozoa. No significant differences between the two freezing media could be identified, suggesting that both cryodiluents would be suitable for incorporation into a sex-preselection protocol for stallion spermatozoa. Experiment 2 compared Kenney's modified Tyrode's (KMT) and Sperm TALP (Sp-TALP) as the staining and incubation medium for stallion spermatozoa prior to sex-sorting. A significant increase in the percentage of acrosome-reacted spermatozoa occurred after staining and incubation in the clarified Sp-TALP compared with KMT. As no improvements in sorting rates were achieved using Sp-TALP, it was concluded that stallion sorting protocols could include KMT as the staining and incubation medium while either INRA 82(®)  or lactose-EDTA could be employed as a cryodiluents.
authors: Faculty of Veterinary Science, University of Sydney, Sydney, Australia.
authors: J R
authors: JR
authors: Clulow
authors: H
authors: H
authors: Buss
authors: G
authors: G
authors: Evans
authors: H
authors: H
authors: Sieme
authors: D
authors: D
authors: Rath
authors: L H A
authors: LH
authors: Morris
authors: W M C
authors: WM
authors: Maxwell
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000662:veterinary; D002470:Cell Survival; D004396:Coloring Agents; D015925:Cryopreservation / Q000662:veterinary*; D003451:Cryoprotective Agents; D005260:Female; D006736:Horses; D006358:Hot Temperature; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D012743:Sex Preselection / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology*
chemical_list: D004396:Coloring Agents; D003451:Cryoprotective Agents