The immunological sperm separation method is economical compared to the existing sorting method, and it is promising for the development of new technologies by reducing sperm damage. Wholemom (WM) is a sex-regulating protein that comprises on immunoglobulin G coupled with magnetic nanoparticles (MNPs) that responds to surface proteins derived from the Y chromosome in cattle. Y sperms are restricted in motility as the WM aggregates them, and the magnet could separate the non-aggregated cells. This study was conducted to investigate the effect of WM treatment on the characteristics of bull sperm. After treating sperm with WM and incubation for 6 h, the motility parameters including total motility, progressive motility, velocity average path, velocity straight line, amplitude of lateral head displacement, and linearity were significantly higher in the WM treatment group than in the control group (p < 0.05). Sperm viability and acrosome reaction rates were similar in both groups during each incubation period (p > 0.05). In conclusion, the immunological sperm sexing procedure using a monoclonal antibody conjugated with MNPs did not affect the characteristics of bull sperm. This study suggests that compared to other techniques, the immunological method for sperm sexing could classify sperm quickly and efficiently without the use of expensive equipment.

본 연구에서는 한우 암소와 씨수소의 육종가를 기반으로 연도별 유전적 개량량을 추정하여 현재 한우 축군의 개량 정도를 파악하여 향후 한우 개량 방향을 위한 기초자료로 활용하고자 본 연구를 시행하였다. 본 연구를 위하여 한우농가 4,040호에서 축산물품질평가원에 도축된 970,567두의 도축자료를 이용하여 10번 반복되어 추출된 표본 자료와 한우농가에서 2009년부터 2019년까지 사육되고 도축된 개체 중에서 한국종축개량협회에 혈통등록 되어 있는 거세우를 선별하여 결측치 및 이상치를 제거하고 도축월령이 27~32개월 이외의 기록은 분석에서 제외한 후, 1,391,141두의 도체 자료를 분석에 이용하였다. 이용된 형질은 도체중(Carcass Weight, CW), 등심단면적(Eye Muscle Area, EMA), 등지방두께(Backfat Thickness, BF) 및 근내지방도(Marbling Score, MS)의 4개 형질을 고려하였다. Random Sampling 10 반복되어 추출된 Data 1의 도체중, 등심단면적, 등지방두께 및 근내지방도의 개량추세에 대한 회귀계수는 매년 평균 0.44㎏, 0.197㎠, -0.051㎜ 및 0.034점으로 나타났으며, 도축성적을 보유한 전체 개체의 자료로 구성된 Data 2에서는 각각 0.35㎏, 0.22㎠, 0.06㎜ 및 0.04점의 개량추세를 보였다. Data 2에서 씨수소의 개량추세는 1.54㎏, 0.343㎠, -0.045㎜ 및 0.050점의 추세를 보였으며 암소와 씨수소 간에는 1.19㎏, 0.119㎠, -0.014㎜ 및 0.010점의 차이를 보였다. 유전적 개량량을 확인하기 위하여 육종가의 표준편차를 이용하였으며, 암소의 선발강도는 본 연구에서 보고한 암소의 후대 거세우 기록 빈도 및 비율을 통해 계산하였다. 한우 암소의 선발 시기를 3산차 이전을 기준으로 하는 것과 4산차 이후를 기준으로 하는 것으로 나눠서 선발강도를 적용하였다. 그 결과, 암소의 선발 시기를 3산차 이전으로 했을 때 세대당 유전적 개량량은 도체중, 등심단면적, 등지방두께 및 근내지방도에서 각각 5.04㎏, 1.31㎠, 0.71㎜ 및 0.32점으로 나타났다. 이러한 결과를 바탕으로 현재 한우 암소집단에서 정확한 육종가를 추정하고, 유전적 개량량을 높이기 위해서는 후대의 기록을 높이기 위한 육종 계획이 필요할 것으로 판단된다.

In the present study, we examined the effect of straw size on spermatozoa motility, viability, acrosome integrity, mitochondrial membrane potential, and plasma membrane integrity after freezing-thawing. Hanwoo semen was collected from three bulls and diluted with an animal protein-free extender, divided into two groups, namely, 10 million spermatozoa in 0.25 mL and 20 million spermatozoa in 0.5 mL straw, and cryopreserved. In Experiment 1, the motility and motility parameters of the frozenthawed spermatozoa were evaluated. After freezing-thawing, the spermatozoa motility parameters fast progressive, straight line velocity, and average path velocity were compared between the 0.25 mL straw and 0.5 mL straw groups. They were 35.2 ± 1.0 and 32.3 ± 0.7%, 34.6 ± 0.7 and 31.8 ± 0.5 μm/s, 51.4 ± 1.3 and 47.1 ± 1.1 μm/s, 0.25 mL straw and 0.5 mL straw groups, respectively. In Experiment 2, the viability, acrosome membrane integrity, and mitochondrial membrane potential of the frozen-thawed spermatozoa were assessed. After freezing-thawing, the percentages of spermatozoa with live, intact acrosomes and high mitochondrial membrane potential were compared between the in 0.25 mL straw and 0.5 mL straw groups. They were 48.0 ± 2.6% and 35.6 ± 2.8% between the 0.25 mL straw and 0.5 mL straw groups. In Experiment 3, the plasma membrane integrity of frozen-thawed spermatozoa was compared. After freezingthawing, the plasma membrane integrity was higher for the in 0.25 mL straw group than the 0.5 mL straw group. They were 62.0 ± 2.2 and 54.1 ± 1.3% between the 0.25 mL straw and 0.5 mL straw groups. In conclusion, our results suggest that freezing semen in 0.25 mL straw improves the relative motility, viability, and acrosomal, mitochondrial membrane potential, and plasma membrane integrity of Hanwoo bull spermatozoa.

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.

노루궁뎅이균사로 발효한 율무 열수추출물의 항산화, NO 생성저해, tyrosinase 합성 억제, melanin 생성 저해 및 유용성분 분석을 수행하였다. 노루궁뎅이균사발효율무 열수추출물의 ergosterol 함량은 740.2 mg%으로 나타났으며, 원료로 사용한 율무 열수추출물에서는 검출되지 않았다. 노루궁뎅이균사발효율무 열수추출물의 β-glucan 함 량은 245.3 ± 5.1 mg%으로 나타났다. 노루궁뎅이균사발 효율무 열수추출물은 100%, 104.1%, 107.2%, 105.3%, 102.1%, 101.3%, 100.4%의 세포생존율이 측정되어, 율무 열수추출물보다 노루궁뎅이균사발효율무 열수추출물의 세포생존율이 더 높게 나타났다. B16F10 cell 500 ug/mL 농도로 처리했을 때, 노루궁뎅이균사발효율무 열수추출물 은 8.9 μM의 NO 생성율을 보여, 율무 추출물(10.6 uM) 의 NO생성율 보다 낮은 NO 생성율을 나타내었다. 10, 30, 50, 100, 200, 300 및 500 mg/mL 모든 농도에서 율무 열수추출물보다 노루궁뎅이균사발효율무 열수추출물이 유의적으로 tyrosinase 및 melanin 생성을 억제함을 확인 할 수 있었다. 노루궁뎅이균사발효에 따른 DPPH radical 및 ABTS radical 소거활성 측정결과 노루궁뎅이 균사발효 율무열수추출물이 율무 열수추출물보다 항산화 효과가 높음을 확인하였다. 따라서 추가적인 연구를 통해 노루궁뎅이균사발효 율무열수추출물은 기능성식품 원료와 식품 첨가물, 화장품 산업에 이용될 수 있을 것으로 생각된다.

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.

In this study, two epididymal spermatozoa recovery methods in relation to sperm number, motility, viability and acrosome reaction were examined. Seven bulls were castrated and 7 testicles with epididymides were transferred to the laboratoy. Epididymis in each bull was randomly used for flushing and mincing methods with semen extender (Optixcell, IMV, France). The recovered spermatozoa with adjusted sperm concentration to 40 × 106 cells/mL was diluted with optixcell and cryopreserved. In experiment 1, the difference in the total number of spermatozoa using flushing and mincing methods was insignificant (2570.0 and 2505.2 × 106 cells/mL, respectively). For experiment 2, the percentage of motile spermatozoa and motility parameters between flushing and mincing methods were studied through the use of sperm class analyzer after frozen-thawing. The percentage of total motile sperm between flushing and mincing methods was almost the same with 89.5±12.8 and 91.4±7.9%, respectively. The same is the case with experiment 3 wherein the viability and acrosomal integrity of frozen-thawed epididymal spermatozoa by flushing and mincing was insignificantly different. The results from the study showed that both flushing and mincing methods can be used for epididymal spermatozoa recovery in bull.

본 연구는 티모시 건초와 농후 사료 위주의 사료를 급여한 한우 씨수소 정소상체 정자 체외수정 효율 조사를 통해 정자의 활용 가능성을 조사하였다. 농후 사료는 체중의 1.8%를 급여하고 양질의 티모시 건초를 자유채식 시킨 14개월령 거세우의 정소에서 분리된 정소상체 미부의 정자를 회수하고 동결 흉해 후 체외수정을 실시한 결과는 다음과 같다. 웅성전핵과 자성전핵이 형성(2PN)된 난자는 정상수정으로, 1개의 전핵(1PN), Expanded Sperm Head (ESH), Polyspermy 형태는 비정상적인 수정의 형태로 평가하였다. 정상적으로 수정된 난자의 비율은 정소상체 정자의 경우 전체 침투율은 49.7% 그리고 정상적인 2PN을 가진 난자는 18.5%를 보였으며, 대조구 정자의 전체 침투율은 54.4%로서 정소상체 정자 보다 높은 결과를 보였으나 유의적인 차이를 보이지는 않았다. 정상적으로 2PN을 형성한 비율은 36.7%로서 정소상체 정자를 이용한 정자 보다 높았으나 유의적인 차이는 없었다. 체외수정 후 발달률 조사에서 정소 상체 정자의 분할률은 81.2%, 대조구 정자는 82.7%로 유사한 결과를 보였으나, 배반포 발달률은 정소상체 정자 24.4%와 대조구 정자 12.2%로 정소상체 정자를 사용한 난자의 발달에서는 유의적으로 높았다(p<0.05).

The Jeju Black Cattle (JBC) are a type of traditional Korean native cattle with a characteristic black fur that covers the entire body. Semen analysis is the most commonly used procedure to evaluate male fertility potential. This study was to evaluate the quality of 10 JBC bulls belonging to Jeju Special Self-Governing Province Promotion Institute. [JBC A∼J grade]. The freezing medium (20% egg yolk plus 20% triladyl) was added in semen sample to a final concentration of 100×106 sperm/ml. For sperm cooling, diluted semen was filled in 0.5 ml plastic straws and then kept in refrigerator at 4°C for 2 h. They were placed in 7 cm over liquid nitrogen (LN2) vapor for 10 min and then directly plunged into LN2 for storage. Thawing was done by transferring the frozen straws into water bath at 37°C for 30 sec for analysis. The sperm motility, vitality and morphology in each group was assessed using the Sperm Analysis Imaging System (SAIS Plus; Medical Supply Co, Ltd., Korea), eosin-nigrosin stain and diff-quik kit. There was no difference in the motility of the fresh groups (87.4 ~ 100%), while it was difference in the frozen-thawed groups (42.8 ~ 98.6%) (p<0.05). The best motility was shown in JBC-B (100/fresh and 98.6%/frozen-thawed). There was significant difference in the vitality of the fresh group (19.8 ~ 59.2%) and frozen-thawed group (21.2 ~ 49.8%)(p<0.05). The highest vitality was also shown in JBC-B (59.2/fresh and 49.8%/frozen-thawed). Morphologically, in fresh semen the highest normal ratio was indicated in JBC-E (90.9%) and in frozen-thawed group the highest was in JBC-C (90.2%). These results demonstrated that the analysis including motility, vitality and morphology of fresh or frozen-thawed semen is valuable to select the high quality sperm using for reproduction.

In this study, we examined number, motility and plasma membrane integrity of spermatozoa from six regions of epididymis in bull. Six testicles with epididymides were castrated from six bulls (mean±standard error, age of days = 441.3±9.6, body weight (kg) = 367±8.4, scrotal circumference (cm) = 30.7±0.4) at Hanwoo Research Institute, NIAS and transported to laboratory within 1 hour. Testicular weight, length, width and circumference were recorded. Epididymis in each bull was randomly used for recovery of spermatozoa. Epididymis was divided into six regions: efferent duct (ED), caput, corpus, proximal cauda (Pcauda), distal cauda (Dcauda) and vas deferens (VD). In experiment 1, we examined sperm number of each region of epididymis. Each region of epididymis contained different number of spermatozoa: ED (37.8±15.7 × 106cells/ml, 8.2%), caput (93.6±18.8 × 106cells/ml, 20.2%), corpus (33.0±8.5 × 106cells/ml, 7.1%), Pcauda (104.2±23.5 × 106cells/ml, 22.5%), Dcauda (180.5±32.5 × 106cells/ml, 39.0%) and VD (14.0±5.0 × 106cells/ml, 3.0%). In experiment 2, sperm motility of each epididymal region was examined by computer assisted sperm analysis (SCA, MicroOptic) system. Sperm motility was divided into 4 groups (fast progressive, slow progressive, non-progressive and immotile) based on WHO guideline. Percentages of fast progressive of Pcauda and Dcauda (11.0±2.3 and 15.4±3.6%) were significantly higher than that of ED, Caput, Corpus and VD which is 0.1±0.1, 1.5±0.6, 1.9±0.7 and 0.3±0.2%, respectively (p<0.05). In experiment 3, percentage of intact plasma membrane spermatozoa of each regions were examined by hypoosmotic swelling test. Percentages of intact plasma spermatozoa were not significantly different among six regions of epididymis: ED, caput, corpus, Pcauda, Dcauda and VD which is 68.0±8.6, 74.0±5.3, 68.5±6.2, 70.8±5.5, 71.0±5.8 and 64.6±10.8%, respectively. In conclusion, in the present study, we found out distribution, motility and plasma membrane integrity of spermatozoa from six regions of epididymis in Hanwoo bull. These results will be contributed to basic research about spermatozoa transportation and characters in epididymis of bull.

In this study, we examined total number, motility and plasma membrane integrity of epididymal spermatozoa from cauda epididymis of bull after preservation at 4ºC. Totally, 23 testicles were castrated from 23 bulls (mean±standard error, age of days = 426.0±7.3, body weight (kg) = 379.7±8.4, scrotal circumference (cm) = 31.0±0.4) at Hanwoo Research Institute, NIAS, and transported to laboratory and preserved on 1, 4 and 6 days at 4 ºC. As control, epididymal spermatozoa recovery from 7 testicles was conducted after transportation to laboratory immediately. In experiment 1, we compared total number of spermatozoa among groups. Total number of spermatozoa from epididymis was not significantly on different preservation day of 0, 1, 4 and 6 which is 1778.0±304.7, 1824.8±343.9, 1228.4±91.7, 1201.8±178.6×106 cells/ml, respectively). In experiment 2, we examined spermatozoa motility and motility parameters (VCL (μm/s), VSL (μm/s), VAP (μm/s), LIN (%)) by computer assisted sperm analysis (SCA, MicroOptic) system. Percentage of motile on 0 and 1 day (88.9±5.2 and 85.8±6.1) was significantly higher than that on 4 and 6 days (32.6±6.5 and 34.3±8.25). Percentage of VCL (μm/s) on 0 and 1 day (93.5±7.6 and 83.0±14.9) was significantly higher than that on 4 and 6 days (36.6±5.1 and 39.5±5.5) (p<0.05). Percentage of VSL (μm/s) on 0 day (28.0±2.1) was significantly higher than that on 1, 4 and 6 days (20.2±3.0, 9.0±2.0 and 8.5±1.6, p<0.05). Percentage of VAP (μm/s) on 0 and 1 days (49.4±3.8 and 41.3±6.6) was significantly higher than that on 4 and 6 days (18.2±3.0 and 19.3±2.8, p<0.05). Percentage of LIN (%) on 0 day (30.7±2.6) was significantly higher than that on 4 and 6 days (23.4±2.7 and 21.1±1.0, p<0.05). Motility of spermatozoa was divided into 4 groups (fast progresive, slow progressive, non-progressive and immotile) based on WHO guideline. Percentage of fast progressive on day at 0 was significantly higher than that on 1, 4 and 6 days (0, 1, 4 and 6 days vs. 19.8±1.9, 10.2±1.1, 2.6±1.0 and 2.3±1.2%, respectively). In conclusion, cauda epididymal spermatozoa should be recovered within one day after preservation at 4 ºC to recover high quality of epididymal spermatozoa in Hanwoo bull

Antioxidants have been added to cryoprotectant or in vitro culture medium for sperm to reduce the detrimental damage, such as reactive oxygen species. However, curcumin, an antioxidant, shows dual effect on the viability and progressive motility of bovine sperm exposed to hydrogen peroxide. Low concentration of curcumin increases sperm viability and progressive motility, whereas high concentration of curcumin reduces them. This study was performed to identify whether TREK-1 channel is related to low sperm viability and motility induced by high concentration of curcumin. Curcumin reduced TREK-1 channel activity in a dose-dependent manner. TREK-1 channel was expressed in sperm obtained from Korean native bull. Treatment with TREK-1 channel blockers, such as curcumin, fluoxetine, GdCl3, and spadin, significantly reduced sperm viability and motility (p < 0.05). However, TREK-1 channel activators showed different effect; linoleic acid showed an increase in sperm viability and motility, and wogonin did not affect them. These results show that TREK-1 channel is involved in the regulation of sperm viability and motility. In particular, high concentration of curcumin might reduce sperm viability and progressive motility of Korean native bull through blockage of TREK-1 channel.

The study aims to assess viability, acrosomal menbrane, integrity and mitochondria membrane potential of sperm separated using a percoll density gradient(45% and 90%) and swim-up methods using Hanwoo epididymal sperm frozen semen. Briefy, motile sperm separated using a percoll gradient and swim-up. 25 μl of sperm dilution from droplets were transferred to 1.5 mL tube and incubated with fluorescent probes at 39°C in dark as follows. After incubation, 75 μl of 5,5',6,6'-tetrachloro-1,1',3,3'- -JC-1; Mitochondrial Membrane Potential Detection kit, Cell Technology Inc., USA) working solution was mixed with sperm dilution and incubated for 30 min. One μl of Hoechst 33342 (H1339; Molecular Probe, Eugene, OR, USA) stock solution was mixed with sperm dilution and incubated for 10 min. And then, 0.5 μl of propidium iodide (PI) stock solution and 0.5 μl of fluorescein peanut agglutinin FITC conjugate (FITC-PNA; Vector Laboratories, FL-1071) were mixed with sperm dilution and incubated for 8 min. After mixing with fluorescent probes and sperm dilution, 5 μl of stained sperm dilution was mounted on a slide glass and covered with cover glass. More than 200 sperms in a slide glass were counted with × 400 magnification by fluorescent microscope (Eclipse Ci_L, Nikon, JAPAN) and evaluated viability, acrosomal membrane integrity, and mitochondrial membrane potential of sperm with triple band filter (DAPI/FITC/TRITC; Nikon, Tokyo, Japan). Live sperm were stained with Hoechst 33342(blue) and dead sperm were stained with PI (red). Damaged acrosomal membrane of sperm was stained with FITC-PNA (green) and intact acrosomal membrane of sperm was not stained. Both of sperm swim-up method with or without BSA separated to Live intact mitochondria (15.39±4.31 vs, 12.58±3.74, and) without significant difference. and percoll density gradient method also similar (7.29±6.54), swim-up method of sperm preparation appeared to be more efficacious in percentage recovery of motile sperm concentration compared to Density gradient method.

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.

The recovery of epididymal sperm in animals is considered as one of the important tools to preserve high value or endangered species. However, there are no appropriate castrating indicators such as months of age in bull, sperm morphology, and motility, particularly in young Korean native bull (Hanwoo). Therefore, this study aimed to investigate sperm number, morphology, and motility of sperm in the epididymis tail of young Hanwoo bulls at 8 and 15 months of age. After castration, epididymal tails were collected and minced with blades to recover sperm. In experiments 1 and 2, sperm number, morphology, and motility were examined. Total number of sperm and percentage of normal sperm from bulls at 8 months of age was lower than that of bulls at 15 months of age after collection (P<0.05). Percentage of abnormal head, tail, proximal cytoplasmic droplet, dead and damaged acrosome of sperm from bulls at 8 months of age were higher than those of bulls at 15 months of age (P<0.05). In experiment 3, sperm motility from bulls at 8 and 15 months of age were examined before freezing and after thawing. Frozen-thawed sperm at 8 months of age showed low total motility and motile sperm with ≥ 25 μm/sec compared to those at 15 months of age and commercially-used sperm (P<0.05). In conclusion, sperm derived from the epididymal tail of bulls at 8 months of age showed high abnormal morphology and poor motility, which are not adequate for AI and IVF. On the other hand, sperm derived from the epididymal tail of bulls at 15 months of age showed high normal morphology and motility.

The objective of this study was to investigate the efficiency of nicotinic acid during in vitro fertilization (IVF) in frozen-thawed bull sperm . The ejaculated semen was diluted with Triladyl containing 20% egg-yolk and cryopreserved in liquid notrigen. The frozen sperm was thawed for 45 seconds in the 38℃ water bath. Sperm was diluted with IVF medium (Bovine-Oviduct medium; BO) containing 0, 15, 30 and 60 mM nicotinic acid (NA), which were incubated at 39℃, 5% CO2 for 0, 0.5, 1, 2 and 4h. The characteristics of frozen-thawed sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI for outer acrosomal membrane damage and Rhodamine123/PI for mitochondrial integrity using flow cytometry. And the sperm ability was analysed by Coomassie brilliant blue (CBB) staining for acrosome reaction state and Rose bengal staining for abnormality. Acrosome reaction and abnormality were analyzed using a microscope. In results, sperm viability was significantly higer in 30 mM group than 0 and 15 mM groups at 1 and 2 h (p<0.05). Outer acrosomal membrane damage was significantly lower in 30 mM group than 0 and 15 mM groups at 1, 2 and 4 h (p<0.05). And mitochondrial integrity was significantly higher in 30 mM group than 0 and 15 mM groups at 2 and 4 h (p<0.05). Also, acrosome reaction was significantly lower in 30 mM than 0 and 15 mM groups at 1 and 2 h (p<0.05) and abnormality was lower NA groups than 0 group at 1 h (p<0.05). In couclusion, we suggest that using the thawing medium containing NA for sperm dilution can be benefical for IVF in bulls

본 연구에서는 한우 보증씨수소 KPN 493, 586, 614, 632, 666을 암소에게 인공 수정하여 생산된 자손 축 총 60두(5처리×12두씩/처리구당)에 대한 육질의 이화학적 특성과 상관관계를 분석하였다. 등심 면적 및 근내지방도는 KPN 586 자손축이 가장 우수하였으며(p<0.05), 육질의 이화학적 특성에서는 처리구 간에 거의 차이가 없었으나 특히 풍미와 육색의 황색도(b값)가 KPN 493이 가장 좋았다(p<0.05). 지방산중 올레인산은 KPN 493은 가장 높았다(p<0.05). 또한 각 형질별 상관관계에서, 도체중과 등심 단면적, 등심 단면적과 근내지방도에서 정(+)의 상관관계를 나타내었고(P<0.01), 보수력은 수분과 조단백질 함량에서 부(-)의 상관관계를 나타내었 (P<0.01). 또한 연도는 다즙성과 정(+)의 상관관계, 풍미는 다즙성, 연도 간에 정(+)의 상관관계, C18:1n9과 C20:1n9와 불포화 지방산 간에는 정(+)의 상관 관계를 나타내었다(P<0.01).