Determination of the number of motile sperm within an ovine semen sample using resazurin
1999
Martin, L.M. | Crenshaw, C.C. | Dean, J.A. Jr | Dart, M.G. | Purdy, P.H. | Ericsson, S.A.
Semen samples were collected from 67 post-pubertal Rambouillet rams using electro-ejaculation. Only ejaculates containing a volume of greater than or equal to 0.5 ml were tested (n = 121). Prior to addition of the dye, a microscopic semen evaluation was performed to assess the number of motile sperm ml(-1) using a Makler counting chamber and a 250 x phase contrast microscope. Resazurin (3.1 microgram) was added to each 0.5 ml sample and incubated for a period of 15 min or unit a visible dye reduction from blue to pink could be detected. Samples were maintained at 37 degrees C through out the test. All ejaculates were identified as having at least the minimum average number of motile sperm ml(-1) (greater than or equal to 1.4 x 10(9)), or below the minimum average number of motile sperm ml(-1) (<1.4 x 10(9)). The sensitivities, specificities, positive (NPV) predictive values and overall accuracies were determined for all samples. The highest sensitivity was 77% at 15s and the highest specificity 100% within 13 min. The highest PPV recorded was 93% at 2.25 min and the greatest NPV was 77% at 2.25 min. The highest overall accuracy was 80% noted at 2.25 min. The results suggests that the reducible dye resazurin is capable of assessing the viability potential of an ovine ejaculate based on the number of motile sperm ml(-1).
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