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After the assessment, each bull ejaculate ( n = 9) with semen volume 4 ml, sperm cell concentration 1.0 10 9 /ml, progressive motility 75%, and sperm viability 80% were split into aliquots according to the experimental design and applied for the following experiments ( Optimization of SeNPs concentration for semen cryopreservation The basic semen extender used in this study was a Tris-based extender as distilled water supplemented with 28 mM fructose, 104 mM citric acid, 297 mM Tris buffer, 5.0% (v/v) glycerol, and 20% (v/v) egg yolk