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Nondialyzable Skimmilk in Diluents for Ram and Bull Semen

Department of Animal Science, Cornell University, Ithaca, New York 14850

ABSTRACT

Bull and ram spermatozoa were stored in synthetic media containing various skimmilk preparations. The reduction in motility of ram spermatozoa diluted 80-fold and incubated at 37 C in synthetic media could be prevented by the inclusion of .8% (w/v) of a lyophilized preparation of nondialyzable skimmilk heated to 92 C for 10 min. Bull spermatozoa incubated at 37 C in buffered saline solutions 60, 80, and 100% of the osmolality of 154 mM sodium chloride, and to which 0, .75, 1.5, 2.25, and 3% (w/v) of the milk preparation were added, survived best in the 80% solution. The detrimental effect of osmolality on sperm motility diminished as the concentration of the milk preparation increased. For either ram or bull spermatozoa, fresh nondialyzable skimmilk was best detoxified by heating after dialysis or by adding .5 or 1.0 mg of cysteine-HCl/ml before dialysis. Dialysis removed excess unbound cysteine-HCl. When the milk preparations were lyophilized for storage and reconstituted as diluents, bull sperm motility was higher during incubation at 5 or 37 C in heated milk preparations than in preparations treated with cysteine-HCl before dialysis. The latter treatment was improved by our removing a precipitate formed when cysteine-HCl was added, but heat treatment still was superior. We concluded that heating and cysteine-HCl addition act differently in detoxifying milk. The most suitable methods of preparing fresh milk diluents may not be satisfactory if the preparations are to be lyophilized for storage before use.

¹ Present address: Wellcome Institute of Comparative Physiology, The Zoological Society of London, London, N. W. 1, England.