A New Diluent for Bovine Semen

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Abstract
Semen has been done in Europe and the United States. The use of egg yolk lecithin in a diluent for semen by lYIilovanov and Selivanova (2) in Russia, and the use of egg yolk by Phillips and Lardy (4) of Wisconsin has proved helpful in keeping sperm cells viable for some time. The work of 1Viilovanov (2, 3) and Phillips and Lardy (4) suggested an investigation of a combina- tion of gelatin, egg yolk, buffer salts and water, as a diluent material in the artificial insemination studies in progress at the Ohio Agricultural Experi- ment Station. It is believed that gelatin (Knox) tends to hold sperm inactive, assists in keeping the particles of the egg yolk and the sperm in suspension, supplies extra nutrients, and retards general contamination (bacteria and molds) during storage. Preliminary work done at this station with the gelatin and the non-gelatin diluents has given encouraging results in favor of the gelatin. Two series of samples have been studied. In the first series 12 samples of bovine semen were diluted four times with a diluent containing 2.14 gin. of gelatin (Knox), 0.2 gin. of KIt2P04, 1.325 gin. of Na~HPQ. 12 H20, 100 cc. of sterilized distilled water, and 100 cc. of fresh egg yolk. The non-gelatin group con- sisting of five samples of bovine semen were diluted four times with a diluent containing 0.2 gin. of KH2PQ, 1.325 gin. of Na2HPQ" 12 H20, 100 cc. of sterilized distilled water, and 100 cc. of fresh egg yolk. The materials other than the egg yolk of both diluents were first dissolved in the water before adding the egg yolk. The range in pH was from 6.7 to 6.85. The vials containing the diluted samples were wrapped in cotton and placed in a refrigerator maintained at 4 ° to 6 ° C. Periodical examinations were made with a microscope after a drop of the diluted semen was placed on a glass slide and warmed to 37 ° C. The gelatin dilutions maintained some sperm motility for an average of 21½ days (range 18 to 30 days), whereas the non- gelatin mixture maintained some sperm life for an average of 14½ days (range 14 to 16 days). The technique of handling the semen in the second series was as follows : Collection and dilution of the semen was carried out under sanitary condi- tions; diluted samples were gradually cooled from 30 ° to 5 ° C. for storage purposes; and periodical examinations were made of the stored samples with a compound microscope (× 440) after a small portion was rediluted and

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