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Effects of Age of Stain, Hydrolysis Time, and Freezing of the Cells on the Feulgen-DNA Content of Bovine Spermatozoa

Department of Dairy Science, University of Illinois, Urbana

ABSTRACT

Feulgen-stained bull spermatozoa were measured cytophotometrically in factorial tests of stain aging, hydrolysis time, semen freezing, and sample difference. None of the treatments significantly affected the area of the sperm nuclei. Regressions of optical density on nuclear area were negative and highly significant. Within one subclass, 51% of the variation in optical density could be explained by the variation in nuclear area. Stock solutions of stain prepared alike but used at different ages on the same semen samples yielded highly significant differences. The older solutions produced higher apparent DNA values than the fresher ones. The optimum hydrolysis time was found to be 8 min. Freezing semen smears for short periods did not appear to influence the measured amount of DNA in the cells. Sample differences of < 2% could not be demonstrated as real differences.