50 ESTIMATION OF CHROMATIN ABNORMALITY OF OGYE ROOSTER SEMEN WITH Diff-Quik STAINING

Abstract

The abnormality of Ogye rooster sperm chromatin could be detected by simple sperm staining. In this abstract, a Diff-Quick staining kit was tested for assessment of chicken sperm quality. Using a standard bright-field microscope, Diff-Quik stains can be reproducibly, easily, and routinely monitored with simple staining. The presence of abnormal chromatin staining of rooster sperm was determined by darker stain in head. In the fresh semen, the viabilities of 3 tested Ogye spermatozoa were 93.53, 82.42, and 90.63%, and normal chromatin rates were 87.96, 74.25, and 85.10%, respectively. However, after cryopreservation, the rates of viability of thawed semen were reduced to 69.58, 61.98, and 72.20%, and normal chromatin rate also reduced to 58.91, 48.49, and 63.34%. A significant correlation between live sperm and normal sperm nuclei was 0.875 in fresh semen and 0.513 in frozen semen. After incubation of sperm at 37°C for 5 min, the rates of viability, chromatin normality, and sperm head activity were shown as 90.63 ± 1.28%, 82.44 ± 8.09%, and 66.68 ± 10.29% in fresh semen. However, the rates of thawed semen were reduced to 67.92 ± 7.55%, 56.92 ± 12.15%, and 47.32 ± 5.02%, respectively. The relationship between chromatin normality and sperm head movements in fresh and thawed semen were 0.564 and 0.540, respectively. With these results, the chicken sperm normality could be assessed by the Diff-Quik staining, which could be used for chromatin status of sperm head and activated morphology of live spermatozoa, as a simple and rapid staining method.