Gas-pressure Technique for Preservation of Bovine Spermatozoa

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Abstract
Two studies were conducted, one to determine the effects of 20 psi of CO2 and N2 upon motility and storage life; the other to measure the effects of 20 psi of CO2, N2, or Ar on motility, storage life, and fertilizing capacity of bovine spermatozoa. In the 1st experiment, a total of 39 ejaculates from 7 dairy bulls were standardized to contain 40 X 106 spermatozoa/milliliter of egg-yolk citrate (1:3) diluent. Split-samples were subjected to 20 psi of CO2 or N2 for 60 min. at 20 -22 C and were then stoppered and hermetically sealed at atmospheric pressure prior to storage (4 C). Microscopic evaluation of the per cent motile cells and spermatozoal progressive motility for Day 0, 3, 7, 10, and 14 significantly (P< 0.01) favored N2 and CO2 treatments when compared with controls. In the 2nd experiment, 15 ejaculates from 5 dairy bulls were standardized, treated (CO2, N2, or Ar), and evaluated in the same manner. Per cent motile sperm and spermatozoal progressive motility were significantly (P<0.01) favored in the descending order by Ar, N2, and CO2 compared with controls. Preliminary nonreturn data from 417 inseminations revealed no significant differences (P < 0.01) between the control semen, CO2 or N2 treated semen, or Ar treated semen after storage from 1 to 4 days, 3 to 7 days, and 1 to 14 days, respectively. This technique prolonged spermatozoal motility, in a fluid medium, beyond that obtained by reduced temperature alone.