1. 말레이시아는 젖소 유전자원 개선을 위해 다수의 가축 사육 프로젝트를 지원해오고 있다. 그러나 소비자의 유제품 수요가 빠르게 늘어나는 상황에서 이를 충족시키기 위해 유전자 원과 유제품을 수입에 의존하고 있는 상황이다. 2. 이는 낙농산업 분야에서의 낮은 젖소 번식능력과 개체 수, 비구조화된 번식체계, 수입의존 유전자원, 수익성 있는 낙농산업 모델 부재, 현대적인 유전육종기술 한계에 기인한다고 볼 수 있다. 3. 말레이시아는 향후 정책지원을 통해 낙농산업의 효율성 향상을 진행할 것으로 전망되며, 이러한 상황 속에서 우리나라의 우수한 낙농 유전자원 수출을 위해서는 현지 낙농산업 현황과 검역절차에 대한 이해가 필요하다. 4. 말레이시아 젖소정액 검역 통관을 위해서는 포괄적인 검역조건 대응으로 협의를 통한 검역조건 간소화가 필요하며, 수출 지속성 제고를 위해 국제협력사업과의 연계, 현지 에이전트 발굴을 통한 적극적인 참여 유도, 한류 등을 활용한 현지 홍보활동 방안이 있다.

1. 농촌진흥청 연구과제를 통해 국내 젖소 정액의 수출여건 조사를 제공하고자 우간다 낙농 유전자원 및 인공수정 현황과 수출사례를 통해 국내 젖소 정액의 수출여건 대응 향상 방안을 제시하였다. 2. 우간다의 농업분야는 가용노동력의 70%, GDP의 약 24%를 차지하고 있을 만큼 GDP와 고용측면에 있어 경제의 핵심부문이며 그 중 낙농산업은 축산분야 중 높은 성장률을 보이고 있다. 그러나 동물유전자원과 관련된 전반적인 업무를 총괄하는 국립동물유전자원센터는 수입 유전자원을 정액 형태로 농가에 보급하는 수준에 그치고 있다. 3. 한국은 열악한 기후조건에도 불구하고 세계 3위권의 마리당 일일 우유 생산량을 기록하여 낙농 유전자원의 우수성을 증명하고 있다. 인공수정을 통한 품종 개량 시 유생산 우수 젖소를 효율적으로 확보할 수 있으며, 우간다 농민은 한국 젖소 정액을 활용하여 평균 유량이 어미 소의 2배인 딸소를 생 산하였다. 4. 우간다에 한국 젖소정액 수출은 양국 정부와 여러 관련 기관들의 협조 하에 이루어졌으며 그 과정에서 발생하였던 내용을 토대로 한국 젖소정액의 수출 진입장벽 해소를 위한 방안을 제시하였다.

The study was conducted to evaluate the seminal attributes and cryo-banking of Achhami (Bos indicus) bull semen. Of two Achhami bulls, 8 ejaculates from each bull were evaluated for seminal attributes. For semen freezing and cryo-banking, 4 ejaculates (having ≥2 mL semen volume, ≥75% of sperm motility and ≥1,000 × 106 cells/mL of sperm concentration) from each bull were used. Semen samples were diluted in egg-yolk-tris-citrate extender using a two-step dilution protocol, and were frozen in liquid nitrogen (LN2) vapour in a styrofoam box. The mean semen volume, colour, sperm mass activity, motility, viability, concentration, abnormal acrosome, midpiece and tail and, abnormal head of two Achhami bulls were 4.4 ± 0.5 mL vs. 2.5 ± 0.2 mL, 2.5 ± 0.1 vs. 2.4 ± 0.1, 3.5 ± 0.1 vs. 3.5 ± 0.1, 77.0 ± 1.1% vs. 78.3 ± 1.3%, 94.4 ± 0.5% vs. 91.0 ± 0.6%, 1137.7 ± 73.7 × 106 cells/mL vs. 1060.0 ± 44.3 × 106 cells/mL, 10.2 ± 0.5% vs. 10.3 ± 0.5% and 6.7 ± 0.5% vs. 8.2 ± 0.3%, respectively. The post-thawed sperm motility and viability were 53.0 ± 2.0% vs. 50.0 ± 0.0% and 80.2 ± 0.4% vs. 73.2 ± 0.7%, while evaluating by computer-assisted sperm analysis (CASA) system, the percentage of the progressive motility, fast motility, slow motility, local motility and immotile sperm were 75%, 68%, 7.4%, 16.6% and 8.6%, respectively. A total number of 620 doses semen straw were cryo-banked. Due to the acceptable post-thawed sperm motility and viability recorded, cryopreservation of Achhami semen is hereby recommended so as to preserve the Achhami breed. For further validation, the fertility will be observed from the produced frozen semen.

Artificial insemination technique has been contributed immensely for production of livestock worldwide as a critical assisted reproductive technique to preserve and propagate excellent genes in domestic animal industry. In the past decade, methods for semen preservation have been improved mostly in liquid preservation method for boar semen and freezing method for bull semen. Among many factors affecting semen quality during preservation, reactive oxy-gen species, produced by aerobic respiration in sperm for survival and motility, are unfavorable to sperm physiology. In mammalian cell as well as in the sperm, antioxidant system plays a role in degradation of reactive oxygen species. Magnetized water forms smaller stabilizing water clusters, resulting in high absorption and permeability of the cell for water, implicating its application for semen preservation. Therefore, this review focuses on preservation methods of boar and bull semen with respect to improvement of extender and reduction of reactive oxygen species by using magnetized water and supplementation of antioxidants.

This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of 2×108/ml by doubling in every 30 minutes at 4℃ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to LN2. The presented straws were examined the viability and motility after thawed at 37℃ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm (93.27±1.62%) than frozen-thawed sperm (73.34±3.27%). However, there were no significant differences between fresh and frozen-thawed dead cell rate (7.35±2.63 vs, 13.71±2.85). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different (72.86±2.83 vs, 81.47±2.48), similarly, the dead cell rates was similar (18.41±3.42% and 17.26±4.25). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.

The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under 30℃ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in 0.3℃. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group (43.3±4.7%) was significantly (p<0.05) higher than other treatment groups (Tissue: 16.3±2.7% and Water: 27.5± 3.1%), dying sperm (SYBR+/PI+) in Air treatment group (55.6±4.7%) was significantly lower than other treatment groups (Tissue: 77.6±3.2% and Water: 67.6±3.3%) (p<0.05). Acrosome reaction in Air treatment group (0.2±0.1%) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: 0.7±0.2% and Water: 0.5±0.1%), the acrosome reaction in Air treatment group (28.6±2.8%) within all sperm also was significantly lower than other treatment groups (Tissue: 44.2±1.8% and Water: 36.2±2.0%) (p<0.05). And mitochondrial intact in Air treatment group within live (97.1±0.4%) and all (61.9±3.3%) sperm were significantly higher than other treatment groups (Tissue: 85.2±3.3%, Water: 87.8±2.9% within live sperm and Tissue: 49.28±3.7%, Water: 42.0±3.1% within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.

Alpha-tocopherol as an antioxidant acts in preservation of chilled semen by preserving cell membrane damage from lipid peroxidation. Optimum concentrations of α-tocopherol in egg yolk-citrate (EYC) extender need to be studied in crossbred bull’s semen. Different concentrations of α-tocopherol viz. 0, 1, 2, 4 and 6mg per ml of extender were used. Semen was collected once a week from four bulls used to regular collection, aged 4 to 7 years, weighing 320 to 450 kg, and with body condition score 4 to 4.5 and scrotal circumference 23 to 32 cm. Semen was evaluated routinely and sperm morphology was viewed under light microscope at ×1,000 magnification after fixing with buffered formal saline. Over 90% had normal head, acrosome, mid-piece and tail. Semen was diluted with egg-yolk-citrate extender to produce 15×106 spermatozoa/ml and 0, 1, 2, 4 and 6 mg/ml α-tocopherol were added. The semens amples were kept at 8℃. Sperm motility and viability were examined daily up to 5 days under light microscopy at ×200 magnification. Sperm viability was acceptable (≥40%) up to the 4th day with all concentrations of α-tocopherol and up to the 5th day with 2 mg/ml α-tocopherol. Sperm motility was acceptable (≥40%) up to the 3rd day irrespective of α-tocopherol concentration, and up to the 4th day with 2 mg/ml α-tocopherol. It is suggested that the lifespan of chilled semen may be extended up to 4 days by adding 2mg/ml α-tocopherol.

The decreased fertility is frequently thought to be problem of cattle production. However, studies figure out that number of these problems is related to bull factors especially in artificial insemination setting. Therefore, this study was designed to investigate the fertility status of bull by their estimated relative conception rate of cows that were inseminated by frozen semen from Korean proven bulls. Here we use the non-return rate (NRR) to access the bull fertility whereas, the NRR was define as the proportion of bulls that semen were used to inseminate cows and the number of cows that did not return for another service within 60 days. The data from 54,388 artificial inseminations (AI) were analyzed from 88 KPN semen. The NRRs of highest and lowest fertile bull were 83.81 and 51.33%, respectively. And mean NRR was 68.27%. In comparison to previously reported study, our data shows 17.38% higher NRR and the absolute value of difference in 50%>NRR and 50%<NRR group was 22.17 and 10.51, respectively (p< 0.001). In conclusion, the decreased fertility might consider as key aspect in achieving considerable conception of cows in existing integrated farming system at Korea.

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

The purpose of this study was to examine the effect of growing stages of the Korean Native Striped Bull (KNSB) on the freezability and fertility of frozen-thawed semen. First, we investigated the total motility (TM) and progressive motility (PM) according to the diluent used for semen freezing. Second, we examined the effect of the age of KNSB on semen volume, TM and PM of fresh and frozen-thawed semen. Third, we examined the effect of frozen semen from the different age of KNSB on the fertilization rate, and the artificial insemination pregnancy rate. The diluents used in this experiment were Triladyl and Tris-egg yolk extender (EYE). Semen was collected from 5 KNSB in the growing stage (15 months) and 5 adult KNSB (36 months). When Triladyl or Tris-EYE extender was used for semen freezing, there was no difference of the mean TM and the mean PM. However, the mean TM was significantly higher in Bull No. 1885 than Bull No. 4283 ( <0.05). The mean volume of semen collected from the 15-month-old bulls (2.3 ml) was significantly lower ( <0.05) than that from the 36-month-old bulls (5.0 ml). The mean semen concentration was similar for the 15-month-old ( spermatozoa/ml) and 36-month-old ( spermatozoa/ml) bulls. For the 15-month-old and 36-month-old bulls, the mean TM of fresh semen were 93.7% and 88.3%, respectively, and the mean PM were 97.0% and 88.3%, respectively; the 15-month-old bulls showed a particularly high PM ( <0.05). For the 15-month-old and 36-month-old bulls, the mean TM (56.0% and 58.0%, respectively) and the mean PM (64.0% and 70.7%, respectively) of frozen-thawed semen did not differ. The development rates of embryos after fertilization and the pregnancy rate after artificial insemination using frozen-thawed semen did not differ according to the bull's age. In summary, semen volume differed according to the bull's age, but semen concentration and survival rate, the fertilization rate, and the pregnancy rate did not differ according to the stripe bull's age. Accordingly, semen from bulls in the growing stage can be collected and frozen for the preservation and multiplication of rare livestock.

The objective of this study was to examine effect of ethylene glycol for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing cryoprotectants (7% glycerol and 3%, 5%, 7% ethylene glycol) and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 min, 5 cm for 10 min and 8 cm for 10 min. And then frozen straw was plunged into LN2. Post-thawed sperm motility, viability and membrane integrity were significantly higher in 5% ethylene glycol (72.5±5.00%, 54.88±0.66% and 46.00±2.40%; p<0.05). Motility and viability were similar between 7% glycerol and 5% ethylene glycol. However, the membrane integrity was significantly higher in 5% ethylene glycol (34.69±4.64% vs 46.00±2.40%; p<0.05). The viability and membrane integrity were significantly higher in 5 cm for 10 min and 8 cm for 10 min than 3 cm for 5 min (viability: 55.81±2.94, 55.19±3.34 vs 47.94±3.48%; p<0.05 and membrane integrity: 44.94±3.51, 46.06±2.25 vs 40.38±1.03%; p<0.05). The percentage of capacitated sperm assessed by CTC staining, percentage of F pattern was higher in 7% glycerol, 5% and 7% ethylene glycol, and AR pattern was significantly higher in 3% ethylene glycol. F pattern was significantly increased in 5 cm for 10 min and 8 cm for 10 min (p<0.05), but AR pattern was significantly increased in 3 cm for 5 min (p<0.05).

희소 한우인 칡소의 정액 동결을 위해서 레시딘을 기본 희석제로 하는 AndroMed와 Tris-egg yolk extender를 사용하여 정자의 생존율과 활력 조사를 위해서 본 연구를 수행하였다. AndroMed 희석제를 사용하였을 때 생존율과 활력은 와 의 결과를 보였다. 그리고 Tris-egg yolk extender의 경우는 각각 와 결과를 보여 생존율에서는 Tris-egg yolk 희석제가 AndroMed 희석제를 사용하였을 때보다 유의적으로(p

한우 종모우에 있어 정액 채취시 승가 조건(의빈우 혹은 의빈대)이 정액성상에 미치는 영향을 구명하기 위하여 의빈우 승가 종모우 21두, 의빈대승가 종모우 9두 등 보증 종모우 30두를 공시하여 연구하였다. 정액은 일 간격으로 채취하였으며 채취 당일 1시간 간격으로 총 2회 정액 채취하여 6개월간 정액성상을 조사하였다. 의빈대 승가 종모우가 의빈우 승가 종모우보다 1차 및 2차 정자농도에 있어 유의적으로 높아(p<0.05) 1차 및 2차 총 정자수에서도

This study was undertaken in an effort to select the sire bull in Hanwoo through in vitro fertilization of proven bull semen. It was used for in vitro fertilization that of the 20 proven bull semen with follicular oocytes derived from slaughterhouse ovaries of Hanwoo. The stage of maturation on the time course of bovine cumulus-enclosed oocytes incubated for 24 hours was found the highest(96.4%) than hose of other maturationi time. In vitro fertilization rate of bovine oocytes with proven bull sperm showed from 61.5 to 88.9%. Polyspermy of in vitro fertilized oocytes according to proven bulls were the highest KP 491(61.5%) nothing but KP 486, KP 491 and KP 497.