Effect of Incubation Vessel on the Recovery of C14O2 during Incubation of Bull Spermatozoa with Glycerol–1–C14

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Abstract
The following incubation techniques were investigated: (a) 125 ml Erlenmeyer flasks vs. 20- by 180- mm test tubes as incubation vessels, and (b) incubation at 5[degree]C for 18 hours vs. 37[degree]C for 3 hours. Each vessel contained 3.5 x 109 washed sperm and 5 [mu]c of glycerol-l-C14 suspended in 5 ml of 0.9% saline. Sperm inactivated with 10 N H2SO4 served as controls and did not produce any C14O2- Irrespective of the incubation vessel used, sperm incubated for 3 hours at 37[degree]C produced 62% more C14O2, but showed 23% less uptake of glycerol-1-C14 than did those incubated for 18 hours at 5[degree]C. Regardless of the time and temperatureof incubation, the specific activity of CO2 from sperm incubated in test tubes was only 45% (p<0.01) of that obtained from samples incubated in Erlenmeyer flasks. There was no significant difference in uptake of glycerol-1-C14 by sperm in the 2 types of vessels. The surface area to volume ratio (S/V) was 10.5 times greater in the flasks than in the test tubes and undoubtedly resulted in less gaseous exchange at the exposed surface of the fluid in the test tubes, thereby reducing the amount of C14O2 recovered during incubation. Data for sugar loss and lactate gain showed that twice as much lactate (P<0.05) accumulated in the test tubes as in the flasks, whereas only 38% as much sugar was used. Thus, the larger S/V ratio apparently contributed to the maintenance of a more aerobic condition in the flasks than in the test tubes, thereby resulting in a more rapid turnover of lactate formed from glycerol 1-C14 to C14O2.