Effect of glutaraldehyde concentration and fixative temperature on the number of spermatozoa with normal acrosomes in goat semen
Two experiments were conducted to determine the best fixative solution and the most suitable temperature for fixing sperm cells from goat ejaculates. In Experiment 1, a 6×3 factorial design was used to test 4 glutaraldehyde concentrations (0.5, 1, 2 and 4%) plus 1 treatment that did not contain a fixative (0%) but to which 0.3% sodium fluoride (NaF) had been added to immobilize the spermatozoa; the control treatment contained no fixative or NaF. The 6 treatments were tested with 3 different solvents PBS, Na citrate and BL-1, representing a total of 18 samples per replicate. The fixed samples always provided a significantly higher (P<0.01) percentage of normal acrosomes than the unfixed samples, whether immobilized with NaF or observed inmediately after dilution. In Experiment 2, a 3×3 factorial design was used to determine the effect of the temperature of the glutaraldehyde fixative solution on the number of morphologically normal acrosomes from goat semen samples kept at 3 different temperatures. Our findings indicated that at all 3 fixative solution temperatures (5, 20 and 37°C) there was a significant difference (P<0.01) in the percentage of normal acrosomes. At 5°C, glutarhaldehyde yielded a general mean number of 53.6 normal acrosomes vs 75.1 at 20°C and 83.05 at 37°C. Based on these results, we recommend that the temperature of a fixative solution be established when designing an experiment using goat semen, since the temperature has a significant effect on the number of the normal acrosomes found in a semen sample. © 1992.
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