Sperm recovery from epididymis in animals considered as important tools to preserve high-value or endangered species. However, there are no appropriate castrating indicators of timing for recovery of sperm which can be available to artificial reproduction technologies such as artificial insemination (AI) and in vitro fertilization (IVF), particularly in young Hanwoo bull. Therefore, this study aimed to investigate semen volume, morphology and motility of sperm in epididymis of young Hanwoo bulls at 8, 13, 14, and 15 months of age. About 2 cm of the epididymal tail only was cut and minced using blades. Minced epididymal tail tissues were mixed with semen extender (OptixCell, France, IMV technologies) and sperm were recovered with a cell strainer (100 μm nylon mesh). The number of sperm at 8 months of age was lower than that at 13, 14, 15 months of age in bulls after collection (33.6±27.2 vs. 352.4±39.2, 320.4±113.6 and 422.8±252.4×107cells respectively; P<0.05). After the frozen-thawed sperm the the percentage of abnormal head, tail and dead damaged acrosome did not differ between the ages 13, 14 and 15 months of age in bulls (P>0.05), however, the dead sperm with intact acrosome (DIA), the numbers showed that more than 15 months in 8, 13, 14 months (8.7±4.1 vs. 47.3±12.2, 34.8±14.0, 28.8±8.5, P<0.05). In addition, frozen-thawed sperm at 8 months of age showed low total motile sperm compared to those at 13, 14 and 15 months of age (26.4±8.2 vs. 45.7±29.5, 62.3±41.0, 70.4±15.9%, respectively; P<0.05). In conclusion, sperm derived from epididymal tail at 8 months of age in Hanwoo bulls showed high abnormal morphology and poor motility, which is not adequate for artificial insemination(AI) and in vitro fertilization(IVF). On the other hand, sperm derived from epididymal tail at 13, 14, 15 months of age in bull showed high normal morphology and motility, which may be available for AI and IVF. Epididymal sperm collected from bulls over 13 months is needed for further study whether to use the actual in vitro fertilization.

Semen collection from epididymis has been used as an important alternative tool, particularly on abnormal animals including those with joint disorder, poor sex drive and poor libido. However, previous studies had been focused on only recovery of sperm derived from testes, regardless of fundamental parameters such as age, body weight, scrotal circumference, testicular size (weight, length, width and volume) and semen volume. In addition, there is no precise castrating periods to recover sperm from young Hanwoo bulls’ testis. Therefore, we investigated fundamental parameters in Hanwoo bulls at 7, 8, 14, 15, 16 and 17 months of age, which parameter can be used for sperm recovery from testis. Before castration, body weight and scrotal circumference were recorded. After castration, testes were transported to laboratory and testicular weight, length and width were recorded. About 2 cm of epididymal tail was collected and minced using blades on 100 mm petridish. Minced epididymal tail tissues were mixed with Optixcell (France, IMV technologies) and debris were removed by filteration with 100 um nylon mesh. Seven to 8 months of age in bulls showed low fundamental parameters compared to those of at 14 to 17 months of age in bulls (body weight, 218-258 vs. 330-429 kg; scrotal circumference, 23.4-24.6 vs. 28.8-32.8 cm; testicular weight, 92-104 vs. 222.3-269.9 g; testicular length, 10.9-11.6 vs. 13.4-15.6 cm; testicular width, 4.7 vs. 6.2-6.6; testicular volume, 126.4-136.4 vs. 268.9-357.6 cm3, semen volume, 3.0-4.8 vs. 60.6-68.3 ml, p<0.001). In conclusion, to recover semen derived from epididymis of testis in young Hanwoo bull, these fundamental parameters should be satisfied; 330 kg of body weight, 28.8 cm of scrotal circumference, 222.3 g of testicular weight, 13.4 cm of testicular length, 6.2 cm of testicular width, 268.9 cm3 of testicular volume. In further study, fundamental parameters of 9 to 13 months of age in bull should be investigated to determine more accurate castrating periods for sperm recovery from testis.