Optimization of protocols for Iberian red deer (Cervus elaphus hispanicus) sperm handling before sex sorting by flow cytometry

• Published in: Theriogenology 2017-04, Vol.92, p.129-136
• Main Authors: Anel-López, Luis, Garcia-Álvarez, Olga, Maroto-Morales, Alejandro, Tarantini, Tatiana, Del Olmo, David, Ortiz, Jose Antonio, Martinez, Emilio Arsenio, Roca, Jordi, Vazquez, Juan Maria, Garde, Jose Julian, Parrilla, Inmaculada
• Format: Article
• Language: English
• Subjects: Animals; Cryopreservation; Cryopreservation - veterinary; Deer - physiology; Flow Cytometry - veterinary; Male; Red deer; Semen Analysis - veterinary; Semen Preservation - veterinary; Sex Preselection - methods; Sex Preselection - veterinary; Sex sorting; Specimen Handling; Sperm; Spermatozoa - physiology; Staining and Labeling
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/j.theriogenology.2017.01.023

Currently, sperm reproductive biotechnologies such as sex sorting and cryopreservation are undoubtedly valuable tools for improving the economic and biological efficiency of red deer production systems. In this context, and because of the particular characteristics of this species (extensive exploitation typically far from laboratory facilities), a key goal is to optimize the design of an adequate handling protocol of sperm samples before samples are subjected to sex sorting and cryopreservation procedures to obtain better outputs from the application of these technologies. The main aim of this paper was to design an adequate protocol for Iberian red deer sperm handling before sex sorting by flow cytometry to obtain optimal yields when sex sorting is used in this species. Semen samples from 11 adult males were obtained by electroejaculation during the breeding season. In this study, we tested different protocols for the handling of Iberian red deer spermatozoa before sorting by using different concentrations of sperm (400 or 800 × 106) and adding or not Hoechst 33342 before the transport of samples to the sorting facilities. Based on the results, the most adequate method used to handle samples before sorting was transportation at a high sperm concentration (800 × 106/mL) without Hoechst 33342. These transportation conditions in combination with Hoechst 33342 staining at 5.2 μL/mL once at the flow cytometry laboratory resulted in better (P  0.05) by sperm handling before sorting. Moreover, the sorting yields were compatible with the practical application of these reproductive biotechnologies. •Offspring sex preselection would greatly improve the benefits of red deer production.•To design species-specific protocols is essential for the maximum efficiency of sex sorting procedure.•High sperm concentration is recommended for the transport of samples to be sorted.•Sorting yields compatible with the practical application of this reproductive technology were obtained.