The Use of Dimethyl Sulphoxide, Glycerol, and Reconstituted Skim Milk for the Preservation of Ram Spermatozoa II. The Influence of Diluent Composition and Processing Time During Freezing to ?79°C with Dimethyl Sulphoxide or Glycerol or Both Compounds

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Abstract
Reconstituted skim milk diluents containing dimethyl sulphoxide or glycerol or both substances were used to deep-freeze ram spermatozoa to -79[degree] C. Very poor revival (less than 10%) followed freezing in dimethyl sulphoxide. Small improvements in revival were obtained by adding 2% (w/v) fructose to the diluent and shortening the equilibration time to 10 min. before freezing. The inclusion of 1.5% (v/v) dimethyl sulphoxide in a diluent containing 7% (v/v) glycerol improved revival. Hypertonic diluents were better than isotonic diluents for freezing ram spermatozoa. Replacement of the milk solids by fructose, while the tonicity of the freezing diluent remained constant, did not significantly affect revival. Spermatozoa revived equally well after storage for 5 hr. at 5[degree] C before freezing either in the presence of glycerol (equilibrated spermatozoa) or its absence (aged spermatozoa).

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