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Factors Affecting Immobilization of Bovine Spermatozoa with CO₂ and Their Subsequent Reactivation¹

Department of Dairy Science, University of Illinois, Urbana

ABSTRACT

The immobilizing effects of C0₂ on bovine spermatozoa, followed by reactivation with several pure and mixed gases, were studied in various suspension media. The motility of spermatozoa in undiluted semen declined sharply upon C0₂ gassing. However, complete immobilization was not observed and a gradual increase in the percentage of motile sperm was noted during continued CO₂ gassing. The motility of CO₂-inhibited sperm cells was completely restored upon exposure to O₂, N₂, He, air, or various mixtures of CO₂, N₂, and O₂ containing up to 20% CO₂. Complete immobilization was observed in suspensions containing 10 x 10⁶ cells per milliliter in saline and in the dialyzable fraction of seminal plasma. Immobilization was incomplete in IVT, in IVT containing varying percentages of seminal plasma, and in the nondialyzable fraction of seminal plasma. Recovery of motility upon oxygenation was observed to be optimal where complete immobilization did not occur. The addition of bovine serum albumin to the dialyzable fraction of seminal plasma showed almost the same effect as shown by the nondialyzable fraction. On the other hand, dextran additions did not show any beneficial effects. Results of these investigations suggested that the buffering capacity of the medium, levels of certain electrolytes, and the nondialyzable fraction of the seminal plasma are, at least in part, important factors in maintaining sperm motility under CO₂.

¹ The data presented in this paper were taken from a thesis submitted by K. Koyama to the Graduate School of the University of Illinois in partial fulfillment of the requirements for the Master of Science degree, 1964.

² Present address: Department of Dairy Science, The Ohio State University, Columbus, Ohio.

³ Present address: Ebetsu, Hokkaido, Japan.