The purpose of this study was to investigate the proper feeding level and duration of fermented barley grain feed before harvesting to improve the availability of barley for feed. Trial 1 was to investigate the proper feeding amount of fermented barley grain fermented feed, and we prepared 32 heads (603.4 ± 42.7kg) of 22-month-old Hanwoo steer (603.4 ± 42.7kg) for 8 heads in 4 treatment groups. 48 heads (625.8 ± 13.1kg) for Trial 2 were used for 12heads per 4 treatments, and were reared for about 9 months until 30 months of age. Trial 1 is treated a Control group that feed 10㎏ of concentrate, replaces 10% fermented barley grain feed(FBGF) of the control by (TRT 1) and 20% (TRT 2) and 30%(TRT 3). Concentrate and FBGF fed 9, 2.1 kg, 8, 4.2 kg, 7 and 7, 6.3 kg on each treatments respectively and 1.5 kg of rice straw for forage. In Trial 2, 10% of alternative diets were judged to be appropriate, and the control of 9kg of diets and barley grain fermented feeds were used to determine the appropriate feeding period. The treatments were 3 months before shipment (TRT 1), 6 months (TRT 2) and 9 months (TRT32). Each treatment group had 8 and 2.1 kg of concentrate and barley grain fermentation, respectively. As a feed, rice straw was fed to 1.5 kg. The daily gains per treatment were higher in TRT 1 and TRT 3 was similar to the Control. Body weight and daily gain during the test period were higher in TRT 1 fed 10% barley grain fermented feed. TRT 2 was the highest at 6.13, and TRT 3 was 6.0, which was higher than 5.63 of TRT 1 and 5.5 of Control.

This study was performed to evaluate the effect of heat stress on the status of physiological responses, blood parameter, serum T3 and cortisol, and heat shock proteins (HSP 27, 70, and 90) of Hanwoo cattle. Six Hanwoo steers (242.8 ± 7.2 kg of BW) were housed in the climate-controlled respiration chambers. The experiment consisted of 7 days (control; 0 day) at thermoneutral (air temperature (Ta) of 15oC and relative humidity (RH) of 60%; temperature-humidity index (THI) = 64), and by 3 and 6 days (treatment groups) at heat stress (Ta of 35oC and RH of 60%; THI = 87). Body temperature of each parts (frank, rump, perineum and foot) and rectal temperature elevated in heat stress groups (3 days and 6 days) than the control group (0 day). Respiration rates increased in 3 days and 6 days (88.5 ± 0.96 bpm and 86.3 ± 0.63 bpm, respectively) from 0 days (39.5 ± 0.65 bpm). Feed intake significantly decreased in heat stress groups (3 days and 6 days, 3.7 ± 0.14 kg and 4.0 ± 0.15 kg, respectively) than the control group (0 day, 5.0 ± 0.00 kg). In addition, final BW significantly decreased in heat stress groups (3 days and 6 days, 211.8 ± 4.75 kg and 215.5 ± 3.50 kg, respectively) than the control group (0 day, 240.0 ± 25.00 kg). However, heat stress has no significant effect on blood parameter, serum T3 and cortisol. Nevertheless, heat stress increased HSPs mRNA expression in liver tissue, and serum concentration of HSPs. Despite Hanwoo cattle may have high adaptive ability to heat stress, our results suggested that heat stress directly effect on body temperature and respiration rate as well as serum and tissue HSPs. Therefore, we are recommended that HSPs could be the most appropriate indicators of Hanwoo cattle response to heat stress.

The objective of this study was to investigate the effects of dry-aging and wet-aging conditions on the chemical composition, meat quality traits (color, water holding capacity, cooking loss, WBSF and pH) and shelf-life TBARS (2-thiobarbituric acid reactive substances) and VBN (volatile basic nitrogen) of loin (M. longissimus thoracis), sirloin (M. glutaeusmedus) and top-round (M. semimembranosus) cuts of Hanwoo cows. The dry-aging condition (temperature ℃, relative humidity %) was done for 60 d as 2℃, 65％, 20 d＋2℃, 75％, 20 d＋4℃, 85％, 20 d while, the wet-aging was done for 60 d at 1℃, 85%. Results revealed that moisture content was significantly higher in the dry-aged samples than that of the wet-aged samples at 60 d (p<0.05). The a*- and b*-values were significantly higher in 3 cuts as compared with those treated with wet-aging condition. During the dry-aging time, the WHC significantly increased in the loin and sirloin and they were significantly higher than wet-aging condition at 40 d and 60 d (p<0.05). The WBSF values significantly decreased with increased aging time and there was no significant difference between two aging methods. The TBARS and VBN contents significantly increased with increased aging time for all the wet- and dry-aged cuts, and their levels still fell within the acceptable standard limit. Generally, the TBARS and VBN contents were significantly higher in the dry-aged cuts, probably due to the increased levels of protein and fatty acids oxidations. Based on the results obtained in the present study, it can be suggested that application of dry-aging method can improve the quality characteristics of beef.

본 연구는 한우의 성별에 따른 도체 특성 및 부분육 생산량을 조사하여 도체로부터 고기의 생산수율을 조사하고자 실시하였다. 한우 180두(암소 80두, 수소 29두 및 거세우 71두)를 도축한 다음 가공단계별 중량변화, 도체규격, 대분할 부위 생산량 및 수율, 소분할 부위 생산량 및 수율, 부산물 생산량 및 수율을 조사하였다. 한우의 절식체중은 안소 634 kg, 수소 721 kg, 거세우 754 kg이었으며, 가공단계 별 중량변화는 냉도체중 381.01~467.60 kg, 대분할육 240.79~310.36 kg, 소분할육 208.27~276.47 kg, 뼈 57.23~76.28 kg, 지방 89.19~138.97 kg을 나타내었다. 한우 도체규격은 거세우에서 전장, 후 사분체장, 경장, 요장, 선장, 퇴장, 흉폭, 요폭, 퇴폭, 흉위 및 흉후가 가장 길었다(p<0.05). 도체중량은 생체중의 성별에 영향을 받았으며, 정육율, 지방율 및 뼈수율은 성별에 따라 영향을 받았다. 대분할 정육율은 수소가 등심, 목심, 앞다리, 우둔, 설도, 양지, 사태에서 유의적으로 높은 수율을 나타내었다 (p<0.05). 도체부산물 수율에서 거세우는 가장 낮은 꼬리반골 및 갈비뼈 생산 수율을 나타내었고 (p<0.05), 암소의 사골 및 잡뼈 생산수율이 가장 낮았다(p<0.05). 도체규격과 부분육 생산수율 조사는 도체특성에 따라 부분육 생산량을 예측할 수 있는 자료로 활용할 수 있을 것으로 사료된다.

한우는 도체형질의 개량으로 높은 개량의 효과를 보이고 있다. 특히 한우의 육질은 등급이 높을수록 식감과 맛이 좋다고 알려져 있기 때문에 가격을 결정하는 핵심적인 요소이다. 식육에서 맛을 결정하는 여러 가지 요인 중 크게 작용하는 부분 중 하나가 풍미이다. 식육의 풍미는 열처리를 통해 발현되고 풍미에 중요하게 영향을 미치는 성분은 지방, 탄수화물 및 수용성 비단백질 물질들이다. 이에 본 연구는 풍미에 영향을 미치는 지방산의 특성을 알아보고 높은 가치의 식육 생산을 위한 개량방향을 제시하기 위해 한우 후대검정우 373두의 등심을 채취하여 지방산과 도체형질의 특성을 알아보았다. 도체중, 배최 장근단면적, 등지방두께, 근내지방도의 성적은 각각 383.73 kg, 83.88 cm2, 10.91 mm, 3.89로 나타났고 한우 등심의 oleic acid (C18:1)와 Linolenic acid (C18:3)의 조성비율은 각각 48.08%와 0.11%로 나타났다. Oleic acid의 유전력은 0.726으로 추정되었고 배최장근단면적과 근내지방도와의 표현형 상관이 각각 0.105와 0.141로 추정되었다. 근내지방도와 배최장근단면적의 표현형은 Oleic acid와 정의 상관을 나타내고 있는 것으로 보아 이 두 형질의 개량은 불포화 지방산 함유량의 증가에도 영향을 미쳐 우수한 풍미를 가진 개체가 생산될 확률이 높아질 것이라고 사료된다.

한국에 있어서 한우는 경제 동물일 뿐 만 아니라 중요한 유전자원 중 하나이다. 그러나 현재 한우의 번식 능력 개량을 위한 연구는 매우 미흡한 실정이다. 한우의 번식 능력에 대한 기초 자료를 확보하기 위해 전북지역(정읍시)에서 인공수정 된 암소 총 33,281두에 대한 105,017개의 번식기록(2010-2017)을 이용하여 임신기간(GP), 분만 후 첫 종부기간(CFSI), 분만 후 임신간격(CCI), 수태당 종부회수(NAIPC), 첫 종부 임신율 (FSCR)을 계산하여 분석에 활용하였다. 전체적으로 GP 는 287.602±4.797일, CFSI와 CCI는 각각 77.513±34.417 및 88.433±42.194일 이였다. 또한 FSCR는 62.365%였으며, NAIPC 는 1.411회로 나타났다. 2017년의 FSCR과 NAIPC는 각각 67.497% 및 1.344회로 2013년 62.366% 및 1.402에 비해 유의적으로 증가하였다(p<0.05). 가을과 겨울에는 번식기간이 더 길었으며, NAIPC는 봄과 여름에 비해 유의적으로 증가하는 것으로 나타났다(p<0.05). 3~7산차 암소의 경우 NAIPC가 1.323±0.669~1.339±0.689회로 다른 그룹에의 1.401±0.772~1.446±0.854회 보다 우수한 것으로 나타났다. 반면, 1산차 암소의 경우 FSCR와 NAIPC가 각각 60.217% 및 1.506±0.937회로 가장 저조한 것으로 나타났다. 본 연구 결과 한우의 번식효율은 산차에 따라, NAIPC는 계절에 따라 영향 받는 것으로 나타났으며, 전체적인 번식효율은 과거에 비해 지속적으로 향상되고 있는 것으로 나타났다.

This experiment was conducted to investigate the effect of whole crop rice (WCR) based TMR on growth performance and carcass characteristics of Hanwoo steers. WCR “Yeongwoo”was harvested at yellow ripen stage and ensiled for 60 days. The crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), in vitro dry matter digestibility (IVDMD) and total digestible nutrient (TDN) content was 8.4 %, 28.0 %, 53.8 %, 72.4 % and 66.8 %, respectively. For silage quality, pH was 4.37 and lactic and butyric acid content were 2.84 and 0.04 % in DM. Sixteen Hanwoo steers (8-mon-old) were allocated into either a control (commercial TMR) and WCR-TMR (WCR-based TMR) group. The TMR were fed according to the feeding stage phase: growing (Initiate∼14 month), early fattening (15 month∼21 month) and late fattening (22 month∼30 month). The body weight of control group increased (P<0.05) until early fattening stage, but late growing stage of WCR-TMR group was higher than that of control (P<0.05). Average daily gain (ADG) was significantly greater (P<0.05) in WCR-TMR group (total 0.78 kg/head) compared to control (total 0.66 kg/head) except for late fattening stage. The marketing weight and carcass weight were higher in WCR-TMR group (726 vs 765 kg; 417.8 vs 450.4 kg). The back fat thickness (11.75 vs 13.00 mm), Longissimus dorsi area (88.00 vs 89.88 ㎠) and yield index (65.87 vs 64.30) were not different between the two groups (P>0.05) and also no difference in meat yield grade (A : B : C = 2 : 4 : 2). Marbling score (4.00 vs 4.13), meat color (4.75 vs 4.75), fat color (3.13 vs 2.88), texture (1.25 vs 1.50) and maturity (2.00 vs 2.00) were not significant difference between the two groups and meat quality grade (1++:1+:1:2:3=0:2:4:2:0) was also not different. In conclusion, TMR feeding based on WCR silage showed superiority in carcass yield and ADG compared to control TMR. It is considered that the use of WCR for feed is a necessary option for the substitution of the imported forages and the government's policy for rice production adjustment.

본 연구는 한우 도체형질에서 거세우 자료로 실시한 유전능력 평가결과가 암소 자료로 실시한 평가결 과와 차이가 있는지를 알아보기 위해서 도축 자료를 암소 자료와 거세우 자료로 분리해서 유전모수와 육종가를 추정함과 동시에 동일 형질의 암소 측정치와 거세우 측정치를 각각 다른 형질로 간주하고 각 형질 내에서 두 측정치간의 유전 상관계수를 추정하였다. 도체중, 등지방 두께, 등심 단면적 및 근내지 방도의 유전력은 암소 자료 추정치들이 각각 0.60, 0.46, 0.47 및 0.59 그리고 거세우 자료 추정치들이 각각 0.62, 0.54, 0.41 및 0.58로 도체중과 근내지방도에서는 두 자료의 추정치 간 차이가 없었으나 등지방 두께 유전력은 암소자료 추정치가 거세우 자료 추정치에 비해 8% 더 작고 등심 단면적 유전력은 암소자료 추정치가 6% 더 큰 것으로 나타났다. 그리고 같은 형질 내에서 추정한 암소 측정치와 거 세우 측정치간의 유전 상관계수는 도체중이 0.77, 등지방 두께가 0.79, 등심 단면적이 0.79, 그리고 근 내지방도가 0.87이었다. 한편 각 형질별로 계산한 암소 자료의 추정 육종가와 거세우 자료의 추정 육종 가간 순위 상관계수는 도축우 아비들이 0.45(도체중)에서 0.65(근내지방도)의 범위, 그리고 도축우 어미들이 0.29(도체중)에서 0.40(근내지방도)의 범위로 낮은 편이었다. 이러한 결과는 추정된 자료에 따라 씨수소나 씨암소의 선발 순위가 달라질 가능성이 있음을 시사한다.

These studies were conducted to evaluate developmental competence of follicular oocyte collected from the ovaries of Hanwoo cows with the high offspring meat quality (1++ and 1+ grade). Cumulus oocyte complexes from individual cows were matured, fertilized and cultured using protocols of in-vitro maturation (IVM), in-vitro fertilization (IVF) and in-vitro culture (IVC). The rates of blastocyst development from Hanwoo cows with the offspring meat quality grades of 1++ and 1+ were 18.6 and 21.2%, respectively. The rates of blastocyst development were 26.3, 20.7, 20.7, 17.2 and 31.2% from Hanwoo cows with the meat quality grades of 1++, 1+, 1, 2 and 3, respectively. Fiftyseven transferable embryos were recovered from 11 Hanwoo donor cows (5.2/head) with the high offspring meat quality grades of 1++ and 1+ in vivo, and the pregnancy rate after embryo transfer was 61.1%. In conclusion, these results suggest that in vitro embryo production from the ovaries of cows with the high meat quality grades using individual culture system can be used an efficient method for livestock improvement. In addition, for the successful industrialization of embryo transfer, conception rate should be improved.

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).

본 연구는 한우 보증씨수소 844두를 출생년도를 기준으로 8개 집단으로 분류하고, 각 개체들의 친자확인용 유전자 마커정보를 농협경제지주 한우개량사업소 홈페이지에서 제공 받아 유전적 다양성 및 구조 변화 분석에 활용하였다. 한우 보증씨수소 전체 집단의 대립유전자수(number of alleles)의 평균은 10.54개, 기대 및 관측 이형접합율(Hex, Hob)의 평균은 각각 0.764, 0.773, 다형성 정보량 지수(PIC)의 평균은 0.727 그리고 Fis의 평균은 –0.014로 확인되었다. 한우 보증씨수소 집단을 출생년도 별로 구분한 8개 집단의 유전적 다양성 및 구조 분석 결과, D집단(2005-2004년)의 기대이형접합율(0.777), 관측이형접합율(0.792) 그리고 다형성정보지수(0.740)가 가장 높은 것으로 확인되었다. C집단(2003-2004년)과 E집단(2007-2008년)에서는 기대이형접합율이 관측이형접합율 보다 큰 것으로 확인되었고, 나머지 그룹 모두에서는 관측이형접합율이 기대이형접합율 보다 큰 것으로 확인되었다. 대립유전자 출현빈도를 기반으로 유전적 조성과 구조를 추론하기 위해 STRUCTURE software를 이용하여 분석한 결과 세대가 지남에 따라 특정 유전적 성분의 변화 또는 비중의 증감을 확인 할 수 있었다. 이는 개량 목표를 설정하고 지속적으로 추진되고 있는 개량 사업이 한우 씨수소 집단의 유전적 구조 변화에 영향을 미치고 있음을 확인 할 수 있는 중요한 자료로, 한우 개량 사업의 효율적인 추진을 위해 유용하게 활용 될것으로 사료된다.

The ovum pick up(OPU) technique can be used to produce embryos after in vitro culture of ovarian oocytes, can be used for early securement for effective herd early proliferation and excellent Hanwoo genetic resources, It is attracting attention as a very important technique for establishing technology. In addition to in vitro culture techniques, the number of oocytes retrieved depends on the environment and timing of the OPU. This study was conducted to compare and examine seasonal effect to the differences in the number of recovered oocytes, recovery rate and embryo development rate using Korean cattle kept in animal genetic resource research center by OPU technique. The grade of COCs was evaluated by microscopic examination. Grade A had 3 or more layers of cumulus cell and compact cytoplasm. Grade B had 1~3 layers of cumulus cell and compact cytoplasm. Grade C had 1 layers cumulus cell and compact cytoplasm. Grade D was denuded oocyte and poor cytoplasm. The recovery rate was 47.8±3.4% in summer (June to August) and 51.6±3.8% in autumn (September to October). The number of oocytes was 5.7±0.6 in summer and 5.2±0.7 in autumn. Oocyte grade A and B was 46.2%±6.3% in summer and 51.1±5.0% in autumn. The cleavage rate was 46.1±7.1% in summer and 65.0±11.3% in autumn. Blastocyst development rate was 19.9±9.4% in summer and 29.0±8.7% in autumn. There was no difference the recovery rate of oocytes and the number of embryos between summer and autumn. Cleavage rate and blastocyst rate of autumn was higher than summer. we will investigate to study the appropriate method for the production of Hanwoo embryos and the systematic comparison.

Serum metabolites were analyzed to investigate relationship of pregnancy and non-pregnancy Hanwoo cows. Totally, 251 Hanwoo cows were used in the present study. Grazing was carried out for 5 months in the pasture. In barn feeding, concentrate 3.0 Kg (TDN 68%, CP 14%) and rice straw 6 kg(TDN 50%, CP 6.5%) were fed. For artificial insemination (AI), progesterone-supplying device (CIDR) was introduced to vagina of Hanwoo cows and 2.0 mL of GnRH. One week after introduction, CIDR was removed and 5.0 mL of PGF2α was injected intramuscularly. After 2.5 day, AI was accompanied by a 2 mL of GnRH intramuscular injection and a second AI was carried out 3.5 day. The pregnancy diagnosis was confirmed by rectal palpation about 90 days after AI. Blood samples were collected from the jugular vein after 3 hours of feeding. Analysis of serum metabolites was performed on six types of metabolites: glucose(mg/dl), cholesterol(mg/mL), BUN(mg/dl), AST(U/L), ALT(U/l), and nonessterified fatty acids(NEFA, uEq/L). The metabolic profile test was analyzed by analyzer (Hitachi, 7020, Japan). Pregnant and non-pregnant groups showed serum metabolites as follows. In 60 pregnant group: Glucose 88.9 ± 2.5, Cholesterol 149.8 ± 4.9, BUN 16.9 ± 0.4, AST 99.1 ± 2.6, ALT 35.9 ± 0.9 and NEFA 326.7 ± 15.7. In 43 non-pregnant cow group: Glucose 89.2 ± 3.3, Cholesterol 165.9 ± 4.6, BUN 17.4 ± 0.6, AST 108.9 ± 0.6, ALT 37.8 ± 1.0 and NEFA 419.2 ± 32.8. Cholesterol, AST and NEFA levels in non-pregnant cows were significantly higher than those in pregnant cows (P<0.05). In sum of grazing and barn feeding group was totally 148 cows. Seventy nine pregnant cows showed high glucose and low NEFA levels compared to 69 non-pregnant cows (P<0.05). In conclusion, pregnant group showed high level of glucose and low level of cholesterol, NEFA. Further study needed to obtain more accurate level of metabolites in serum for pregnant and non-pregnant cows.

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 sperm penetration and blastocyst developmental rate of oocytes to determine fertilizability of cauda epididymal spermatozoa in Hanwoo bull. One testicle with epididymides were castrated from one Hanwoo bull (14 months of age) and transported to laboratory. Spermatozoa recovered from cauda epididymis by mincing with semen extender (Optixcell, IMV, France) and cryporeserved in liquid nitrogen tank until use. As control, frozen Hanwoo semen was used. Cumulus oocyte complexes (COCs) were collected from follicles (2-8 mm) of slaughtered ovaries and 10 to15 COCs were matured in 50μl droplet with M-199 media supplemented with 10% fetal bovine serum, 10μg/ml FSH, 10μg/ml LH, 10μg/ml EGF for 22 to 24 hours in a humidified atmosphere of 5% CO2 in air. After maturation of COCs, matured COCs were co-incubated with cauda epididymal spermatozoa in 100μl droplet in modified Brackett and Oliphant media supplemented with 2.5 mM theophylline for 12 or 18 hours under 5% CO2 in air. Sperm concentration was adjusted to 5 × 106cells/ml. After IVF for 18 hours, presumptive zygotes were cultured in modified synthetic oviductal fluid with 1mM glutamine, 12 essential amino acids, 10 μg/ml insulin under 5% CO2, 5% O2 in air. In experiment 1, we examined sperm penetration rate at 12 hours of IVF of frozen-thawed epididymal sperm. Total penetration rate among cauda epididymis and control were not significantly different (mean±standard error, cauda epididymis and control vs. 49.7±11.3 and 54.4±12.8%) In experiment 2, cleavage and blastocyst development rate were evaluated at day 2 and day 8 after IVF for 18 hours. Cleavage rate among cauda epididymis and control was similar different (cauda epididymis and control vs. 81.2±3.4 and 82.7±2.5%). However, blastocyst developmental rate of cauda epididymis group was significantly higher than that of control group (cauda epididymis and control vs. 24.4±1.6 and 12.2±2.8%, p<0.05). In conclusion, cauda epididymal spermatozoa in Hanwoo bull has high fertilizability and embryo development. Cauda epididymal sperm can be used as an alternative to ejaculated frozen sperm in vitro.

The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58 ± 8.31 and 13.25 ± 7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75 ± 1.98 vs. 8.23 ± 6.07, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.