The use of transmigration and Spermac™ stain to evaluate epididymal cat spermatozoa

Epididymal spermatozoa of domestic cats were diluted with TEST medium and frozen. The parameters — estimated percentage of motile spermatozoa, concentration of spermatozoa, cell morphology and transmigration rate (TMR) — were evaluated before freezing and after thawing. Transmigration is a new method to measure the percentage of spermatozoa that consistently move forward, and has not been investigated with cat spermatozoa until now. Estimated percentage of motile spermatozoa averaged 65%, TMR was 76%, concentration of spermatozoa was 30,000 μl−1 and the incidence of morphologically abnormal spermatozoa averaged 58% before freezing. After thawing, the estimated number of motile spermatozoa declined by 22%, but TMR remained at 76%. The TMR did not correlate with estimated motility but mostly was higher than the latter, which is postulated to be caused by the mobilizing effect of the countercurrent in the transmigration apparature. The estimated percentage of motile cells in the target chamber of the transmigration apparature was improved by using phosphate-buffered saline (PBS) as transmigration medium. Morphology was assessed both after fixation of spermatozoa in Hancock solution and after staining of smears with Spermac™. Spermac™ did not stain all protoplasmic droplets but proved to be more suitable for the routine examination of acrosomal morphology after thawing.