Live normal and dead sperm were determined using eosin/nigrosin staining [23]. The dilution rate for staining was 1:4 eosin-nigrosin (5 μL of raw semen and 20 μL eosin-nigrosin staining). The second slide was used to smear semen across the surface of the first slide and allowed to dry at room temperature for approximately 45 - 60 min before evaluation [23]. The slides were placed on a microscope table and a drop of immersion oil was placed on the slide to evaluate live normal or dead, sperm