Journal of Andrology
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Published-Ahead-of-Print May 28, 2009, DOI:10.2164/jandrol.108.007500

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Comparison of Different Methods for Assessment of Sperm Concentration and Membrane Integrity with Bull Semen

Muhammad Anzar *, Tom Kroetsch , and Mary M. Buhr

* To whom correspondence should be addressed. E-mail: muhammad.anzar{at}agr.gc.ca.

Assessing semen quality is crucially important for the exploitation of genetically superior sires in artificial insemination (AI) programs. This study compared modern and conventional techniques to estimate bovine sperm concentration and membrane integrity. First, the NucleoCounter SP-100 was validated for sperm concentration, and provided statistically reliable and repeatable estimates among aliquots and replicates of 25 fresh ejaculates. Sperm concentrations in 78 ejaculates were then determined with hemacytometer, flow cytometer and NucleoCounter SP-100 and were significantly correlated (P<0.001), with regression coefficients among these three techniques close to 1 (P<0.01). However, the sperm concentration determined by hemacytometer was lower (P<0.01) than by flow cytometer and NucleoCounter SP-100. Forty frozen-thawed semen samples were then assessed for sperm concentration and membrane integrity with hemacytometer, flow cytometer and NucleoCounter SP-100. Significant relationships were found for sperm concentration determined by hemacytometer and NucleoCounter SP-100, and for sperm membrane integrity determined by flow cytometer and NucleoCounter SP-100 (P<0.01). Finally, the standard curves of sperm concentrations in six spectrophotometers, comparing optical density against counts drawn by hemacytometer and NucleoCounter SP-100 (n= 94 fresh ejaculates) showed different (P<0.01) intercepts and regressions coefficients (linear, quadratic, cubic). It was calculated that a breeding station can improve its production potential by 13% using NucleoCounter SP-100 instead of hemacytometer for calibration of spectrophotometers. Flow cytometer and NucleoCounter SP-100 can be used with equal confidence to estimate sperm concentration and membrane integrity in domestic animals and human semen.



Key words: Semen Analysis • Flow cytometer • Membrane integrity • NucleoCounter SP-100 • Sperm concentration




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