Effect of glutaraldehyde concentration and fixative temperature on the number of spermatozoa with normal acrosomes in goat semen
Pintado Sanjuanbenito, B. | Perez Llano, B.
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 X 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 immediately after dilution. In Experiment 2, a 3 X 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 degrees C) there was a significant difference (P < 0.01) in the percentage of normal acrosomes. At 5 degrees C, glutaraldehyde yielded a general mean number of 53.6 normal acrosomes vs 75.1 at 20 degrees C and 83.05 at 37 degrees 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.
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