Commercial applications of OPU/IVP were to produce embryos and calves from high genetic cows.The aim of this present study was to compare the number of recovered oocytes and cultured in vitro produced embryos from Ovum Pick-up (OPU). OPU derived embryo production was carried out of oocytes by ultrasonographic guided follicular aspiration and then produced in vitro produced blastocysts by IVP culture system. In result, the rate of recovered oocytes was obtained 612 (57.2%) and 451(73.7) G1+G2 grade oocytes. No difference of recovered rate (51.1~62.1%) was seen in six donor. The rate of cleavage and blastocyst development were obtained 320 (70.9%) and 78 (24.4%) that was 3.3±0.4 cleaved embryo and 0.9±0.2 blastocysts per session. Cleavage rate of OPU oocytes in No. 6 donor was 90.6%, significantly (P<0.05) higher than that in the other donors, However, blastocysts was similar (25.8~30.0%). In conclusion, limited numbers of OPU oocytes had competent development when cultured in SOF culture medium

The present study was conducted to investigate the effect of BHT supplementation on sperm motility, viability, acrosomal integrity and plasma membrane integrity after frozen-thawing. One ejaculate was collected from one fertile Hanwoo bull by using artificial vagina at Hanwoo Research Institute. The ejaculate was transferred to laboratory immediately and diluted with pre-warmed semen extender (Optixcell, France) (1:1). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and divided into 5 groups according to BHT concentration (0, 0.5, 1.0, 2.0 and 4.0 mM) and cryopreserved in LN2 tank until evaluation. Frozen-thawed semen was transferred to 1.5 ml tube and incubated for 0, 2 and 4h. Sperm motility and motility parameters (total motility, VSL with 25μm≥, VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were evaluated by sperm class analysis (SCA, IVOS, Spain). There were not significant effects of BHT supplementation (0, 0.5, 1.0 and 2.0 mM) on total motile and VSL with 25μm≥ at 0, 2 and 4h. However, 4.0 mM of BHT supplementation showed negative effect on total motile (26.3%), VSL with 25μm≥ (1.3%) at 0 h (p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method and divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). There were no significant differences of LIA, LDA, DIA and DDA on various BHT concentrations at 0 and 2 h. However, 4.0 mM BHT supplementation showed decreased LIA compared with 0, 0.5, 1.0 and 2.0 mM BHT at 4 h (34.6, 37.1, 43.6, 45.4 and 14.7% vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.01). Addition of 4.0 mM of BHT showed negative effect on plasma membrane integrity compared with that of 0, 0.5, 1.0 and 2.0 BHT at 2 h (71.9, 64.2, 64.6, 67.5 and 31.7 % vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.05). In conclusion, various BHT concentrations on optixcell extender showed no improvement on sperm motility, viability and plasma membrane integrity.

The present study was conducted to investigate the effect of different heights from liquid nitrogen (LN2) vapor on sperm motility and morphology after frozen-thawing. Two ejaculates were collected from 2 fertile Hanwoo bulls (A and B) by using artificial vagina at Hanwoo Research Institute. After collection, ejaculates were transferred to laboratory immediately and diluted with semen extender (Optixcell, France). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and cooled at 4°C for 4 h and loaded to 0.5 ml straws. The straws were divided into 2 groups. Straws were placed in 3 or 9 cm of LN2 vapor for 14 min and then plunged into LN2 tank and cryopreserved until evaluation. Sperm motility and motility parameters (total motility, VSL with 25μm≥, VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were evaluated by sperm class analysis (SCA, IVOS, Spain) after frozen-thawed. In bull A, 3cm group showed higher percentages of total motility, VSL with 25μm and VAP compared those with 9cm group (98.0 vs. 93.4%, 62.4 vs. 54.0% and 98.6 vs. 93.2%, 3 vs. 9 cm, irrespectively; p<0.001). In bull B, frozen-thawed sperm of 3cm group showed higher percentages of VSL with 25μm, VCL, VSL, VAP and BCF compared with those of 9cm group (43.5 vs. 26.0%, 123.8 vs. 111.6 μm, 62.9 vs. 57.3 μm and 81.5 vs. 72.5 μm; 3 vs. 9 cm, irrespectively; p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). In bull A, frozen-thawed sperm of 3 and 9cm groups showed no significant difference in LIA, LDA, DIA and DDA. In bull B, 3 cm group showed higher LIA and lower DIA compared with those of 9 cm group (73.2 vs. 23.7% and 23.7 vs. 32.2%, 3 vs. 9 cm, irrespectively; p<0.001). We suspected that 3 cm vapor on LN2 vapor might be affected positively spermatozoa viability and acrosomal integrity compared with 9 cm group. In conclusion, semen freezing procedure in the present study will improve sperm quality after frozen thawing.

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.

We investigated and analysed the actual conditions and characteristics of railroad noise levels for 17 sites in the vicinity of the Seoul-Pusan Line. The results are summarized. as follows : 1) Railroad noise level ranged to 64∼74 L_eq dB(A) at day time and ranged to 60∼72 L_eq dB(A) at night time. 2) Increased night noise level depend on the increase of trains passing at night time. 3) The major factor of increased noise level in the vicinity of stations are using loudspeakers and stream whistle on trains. 4) Decreased effect of noise according to distance is able to be described quantitatively using regression equations of multiplicative model. L_eq=78.59 X^-0.056 n=25, r=-0.994, s.e.=1.007 P_av =105.68X^-0073, n =25, r =-0.997, s.e. = 1.007 Also increased and decreased effect of noise according to floor in apartment is able to be described quantitatively using regression equations of multiplicative model. L_eq = 64.238 X^0.0567, n = 39, r = 0.787, s. e. = 1.004 P_av. = 79.963 X^0.0254 n = 39, r = 0.689, s. e. = 1.056 5) Average noise level in high floor is over 70L_eq dB(A) at day and night time, so more detailed soundproofing countermeasured in high floors apartment is required.