This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 porcine FSH, 0.5 equine LH, 1.0 17 -estradiol () and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at in humidified atmosphere of 5% in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.

This study was carried out to investigate the general characteristics, such as volume, pH, sperm motility and sperm concentration of the semen collected from Beagle dogs (age 24～48 months, weight 10～15 kg) by using the method of digital manipulation of the penis, and the effect of preservation temperature and time on motility of fresh semen. Multiple ejaculates were collected from 4 male Beagles. The average volume, pH, motility and sperm concentration of the second fraction (contained with small volume of the third fraction) per ejaculation were 2.94±0.24(SD) ml, 6.43±0.42(SD), 97.04±3.50(SD)% and 1.67±0.23(SD)×108 cells/ml, respectively. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculate were 1.24±0.20(SD) ml, 6.03±0.26(SD), 11.30±4.02(SD)% and 7.25±1.02(SD)×105 cells/ml. Those of second fraction were 2.52± 0.32(SD) ml, 6.32±0.31(SD), 96.25±3.52(SD)% and 2.35±0.35(SD)×108 cells/ml. Those of third fraction were 2.71±0.27 (SD) ml, 6.52±0.20(SD), 95.65±2.78(SD)% and 5.72±0.29(SD)×107 cells/ml. Motility of semen was higher at 17℃ preservation temperature than 5℃ or 36℃ during preservation period. When preservation temperature was 17℃, motility was 96.54±2.05(SD)% at 1 h, 90.20±3.90(SD)% at 6 h, 89.05±2.01(SD)% at 12 h, 78.21±3.50(SD)% at 18 h, 45.24±6.25 (SD)% at 24 h and 30.75±17.24(SD)% at 30 h, respectively.

Prostaglandin () can facilitate release of epinephrine from the adrenal gland. The objective was to extend these findings and determine the effects of on sexual activity and semen collection training in sexually inexperienced boars. Boars (n=32; days of age) were moved individually once weekly to a semen collection room equipped with an artificial sow. Before entering the semen collection room, boar received i.m. treatments of at doses of 5 (n=8), 10 (n=8), or 20 (n=8), and control boar (n=8) were not treated. Reaction time (elapsed time after entering collection pen until the start of mounting) for boars receiving 5mg (), 10mg () was shorter (p<0.05) than for controls (). Duration of ejaculation (min) per session was longer (p<0.05) for (10 mg, 20 mg)-treated boars (, ), compared to control (). The number of training session per boars was less (p=0.056) for 10mg-treated boars (), compared to control (). Semen characteristic such as volume, concentration, the number of total ejaculated sperm, were similar for -treated and controls. There was no apparent difference on sperm movement characteristics (Mot: motility, VCL : curve linear velocity, VSL : straight line velocity, VAP : average path velocity, LIN : linearity) after semen preservation by collected with or without treatment. In summary, administration of in boars increased the sexual activity and facilitated the training boars to mount an artificial sow for semen collection, but did not affect semen characteristic.

We tested the effects of roasted and non-roasted Zizyphi Spinosi Semen (ZSS) hot water extracts on pentobarbital-induced sleeping behavior, pentylentetrazole-induced convulsions, and locomotor activity in mice. Both ZSS extracts decreased locomotor activity and pentylentetrazole-induced convulsions. Both extracts alos dose-dependently shortened the sleep onset and prolonged the sleep time induced by pentobarbital, with roasted extract being more potent. In addition, chemical constituents in HPLC also were different. Thus, both roasted and non-roasted ZSS extracts potentiate the sleep onset and the sleeping time induced by pentobarbital, decrease locomotor activity, and reduce pentylentetrazole-induced convulsions, with roasting improving extract potency.

The techniques for the collection, cooling and freezing of semen and artificial insemination of horses are not fully understood in Korea. We investigated the percentages of total motile (TM) and progressively motile (PM) sperms after the collection, cooling and freezing of stallion semen. The average volume of semen was 167 ml in Thoroughbred and 68 ml in Arab. The average numbers of spermatozoa in Thoroughbred and Arab were and respectively. The average percentages of TM and PM were 82.3% and 88.6% in Thoroughbred, and 61.4% and 82.6% in Arab, respectively. The average percentage of TM at 4 hr after cooling at was significantly lower than that at 0 hr (<), but the percentage of PM was similar between 66.5 and 73.2% at 0, 1, and 4hr. The average percentage of frozen-thawed Thoroughbred semen frozen in MFR5 extender was 56.2%, which was significantly higher than that of the semen frozen in LE extender (average 32.9%, p<0.05). The percentage of TM in Arab was similar for semen frozen in MFR5 extender and LE extender (18.2% and 21.2%, respectively), but the percentage of PM was significantly higher in sperm frozen in MFR5 extender than in sperm frozen in LE extender (69.0% vs. 36.4%, p<0.05). Four mares were artificially inseminated by Thoroughbred frozen-thawed semen and one of them fertilized at 11 day after artificial insemination. In this study, the collection, cooling and freezing of equine semen were possible under domestic conditions.

본 연구는 정액의 보존 기간 동안 정액의 질적 변화를 알아보고자 시행하였다. 돼지 정액을 Beltsville Thawing Solution (BTS)에 희석한 후 17'C 에서 5일 동안 보존하였다. 보존 기간 동안 정자의 운동성(%)과 linearity는 3일째부터 유의하게 감소하였으나, 다른 운동 역학 변수에서는 유의적 변화를 나타내지 않았다. 또한, 5일 동안 정액을 보존할 경우 첨체의 온전성에도 변화가 없었다. 그러나 제 4일째부터 첨체 변화가 야기된 정자는 유의적으로 증가하였으나, 수정능 획득이 일어난 정자는 유의적으로 감소하였다. 정액의 보존 기간 동안 첨체의 온전성의 유의적 변화가 없었다. 즉, 보존 기간 3일동안 정자의 질적 운동성 및 첨체 온전성에는 유의적인 변화가 없었으므로 상업용 돼지 액상정액은 17'C 에서 적어도 3일간 수정능력을 만족스럽게 유지함을 보여준다.

The objective of this study was to determine the effect of administration of Prostaglandin F2α (PGF2α) on semen collection training and semen characteristics in sexually inexperienced boars. Boars were moved individually to a semen collection pen and were trained to mount dummy sow. During the first and second semen collection secessions, 4 out of 17 boars and 4 out of remaining 13 boars allowed collection of semen. The 9 boars that failed semen collection from first 2 attempts received immediately 15 mg of PGF2α i.m. (intramuscular injection) upon entering the collection pen for semen collection resulted in successful semen collection from all 9 boars. Total numbers of spermatozoa were higher in PGF2α treated boars but there was no significant difference in % motility kinematics characteristics between control and PGF2α treated groups during 72 hr period. Overall, administration of PGF2α in sexually inexperienced boars increased the sex drive and facilitated the mounting activity to the dummy sow for semen collection.

The aim of this study was to evaluate the effects of orvus es paste(OEP) on the sperm characteristics during freezing in boar semen. Semen quality was evaluated the motility, membrane integrity, mitochondria function, acrosome status, viability and abnormality. Boar semen were frozen until 5℃ for 2 hours using cell freezer and making the straws, and then freezing by lowing the straws into styrofoam box on the 8cm above the LN2 and plunged into LN2 for cryopreservation. In different concentration of OEP (0, 0.25, 0.5 and 1.0%) into cryo-extender, sperm motility, membrane integrity, acrosome status, viability and mitochondria function were significantly higher in 0.5% OEP than those of any other groups, but sperm abnormality were highest in 1.0% OEP group among all treatment groups (P<0.05). In the relationships of the evaluation methods for sperm viability, CBB vs membrane integrity, CBB vs HO/PI and CBB vs mitocondria function were positively correlated (0.67~0.92). Among the evaluation methods, CBB vs membrane integrity, CBB vs HO/PI and CBB vs mitocondria function were significantly correlated (P<0.001). These results of this study indicate that supplementation of 0.5% OEP in boar semen cryo-extender can improve the semen quality.

본 연구는 0.5 ml straw를 이용한 정자 동결융해 시 융해 온도가 동결정자의 성상에 미치는 영향을 파악하고 미니 돼지의 동결정액에 최적화된 적정 융해 조건을 찾기 위하여 실시하였다. 정액동결 straw를 37, 50 및 70℃에서 각각 5초, 10초 및 45초간 융해하여 생존율(SYBR-14/PI staining), 정자원형질막기능검사 (Hypoosmotic Swelling Test) 및 첨체반응율 (CTC : chlorotetracycline staining)을 검사 한 결과 70℃에서 5초간 융해한 정자의 생존율과 CTC 결과가 37℃와 50℃에서 10초 및 45초간 융해한 처리구보다 유의적(p<0.05)으로 높은 생존율과 낮은 비율의 첨체 반응율을 얻었다. 따라서 미니 돼지의 동결 정액은 고온에서 단시간 융해를 하는 것이 정자의 성상에 유리한 것으로 사료된다.

본 연구는 체세포를 이용하여 생산된 복제 한우 수소의 번식능력을 검토하기 위해 실시하였다. 복제 한우 수소(C-38 및 C-39) 또는 일반 한우 종모우로부터 정액을 채취하여 정자의 수 및 동결 전후의 생존성 등을 살펴보았으며, 정자의 운동성 등은 computer assisted sperm analysis(CASA)를 이용하여 측정하였다. 또한, 이들의 수정 능력을 확인하기 위하여 체외수정과 인공수정을 각각 실시하였다. 정액 성상에서는 복제 수소들과 일반 종모우 간에 정액의 양, 정자의 농도 및 동결융해 후의 생존성 등에서 차이가 나타나지 않았다. CASA를 이용한 분석에서 운동성, 곡선 운동 속도(VCL), 직선 운동 속도(VSL) 및 평균 진행 속도(VAP) 등은 복제 수소의 정액이 일반 종모우의 정액에 비하여 유의적으로 높았다(p<0.05). 체외수정에 따른 수정란의 분화율 및 배반포로의 발달율은 복제 수소와 일반 종모우 간에 차이가 나타나지 않았다. 복제소 정액(C-38)을 이용하여 인공수정을 한 5두의 체세포 복제 대리모에서 암수 각각 한 두씩의 건강한 복제 후대 송아지 2두를 생산하였다. 이상의 결과를 종합하여 보면, 실험에 공시된 복제 수소 개체 간의 차이가 나타나기는 하였지만, 복제 수소는 정액 성상과 정자의 운동성 등에서 일반 종모우와 차이가 없었으며. 또한 인공수정을 통해 송아지를 생산함으로써 정상적인 번식능력이 있음을 확인하였다.