The aim of the study was to produce sexed progeny of the genetically confined White Hanwoo (albinism). A White Hanwoo bull was selected as the semen donor. The X-bearing sperm were sorted by MoFlo XDP cell sorter from Korea Sexing Biotech (Daegu, South Korea). To compare pregnancy and birth rates as well as semen quality, a Korean proven bull (KPN) was used as control. Total number of nonsorted sperm was 20 × 106 per straw. Double insemination of 2 or 4 × 106 sexed and frozen X-bearing sperm were performed on Hanwoo heifers. The abnormality of nonsorted albino spermatozoa (5 ejaculates) was 24.9 ± 7.31% (but 18.7 ± 6.2% C in KPN 768, derived from 5 straws) with a higher number of distal reflex abnormality in midpiece (11.7% compared with that of KPN 768, 5.6%). There were no differences in pregnancy and birth rates when 2 or 4 × 106 X-bearing sperm were inseminated but nonsorted semen had higher pregnancy rates (P < 0.05). The pregnancy rates of KPN 768, 2 × 106 cells, and 4 × 106 cells X-bearing sperm of White Hanwoo cattle were 85.0% (17/20), 26.3% (5/19), and 50% (7/14), respectively. The birth rates were 80.0% (16/20), 15.8% (3/19), and 21.4% (3/14), respectively. The female offspring rates were 43.8% (7/16), 100% (3/3), and 100% (3/3) (P < 0.05). These results indicated that sex-sorted albino Hanwoo semen could be used for the production of wanted progeny with 2 × 106 cells/straw for AI to heifers.
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.
Germplasm cryopreservation from a desired species with agricultural and genetic importance would protect them from the risk for extinction. Semen freezing from Korean native cattle would be a good approach for protecting genetic resources due to their limited numbers. It has been known that sperm could resist cryo-damages by freeze-thaw cycles. Thus, we performed 2 refreezing experiments with different initial thawing temperatures using frozen Korean native cattle semen. A total of 5 Hanwoo, Korean Albino, and brindle cattle were used as semen donors. After thawing by using 5°C/2 min or 37°C/40 s with cooling rates, the semen was diluted with the same volume of cryo-media in the first thawing temperature and refrozen. Sperm motilities were determined and compared between animals and groups after rethawing. The mean sperm concentration and motility was 45 × 106 mL–1 (range 2.3 to 89 × 106 mL–1) and 40% (range 13 to 55%). Mean values of motility and viability of sperm that underwent second preservation were significantly higher in 5°C than in 37°C (P < 0.01). However, the activity of viable sperm thawed at 5°C was significantly decreased before refreezing. It is estimated that refreezing of frozen semen from rare Korean native cattle is possible with resistant properties of survived spermatozoa. The higher motility and viability of refrozen semen could be obtained with 5°C thawing procedure for reuse of frozen semen.
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