Background: Sperm quality and the number of sperm introduced into the uterus during artificial insemination (AI) are pivotal factors influencing pregnancy outcomes. However, there have been no reports on the relationship between sperm concentration at AI and sperm quality in Hanwoo cattle. In this study, we examined sperm quality and pregnancy rates after AI using sperm inseminated at different concentrations. Methods: We evaluated the motility, viability, and acrosomal membrane integrity of sperm at different concentrations (10, 15, 18, and 20 million sperm/straw) in 0.5-mL straws. Subsequently, we compared the pregnancy rates after AI with different sperm concentrations. Results: After freeze-thawing, sperm at the assessed concentrations showed similar viability and acrosomal membrane integrity. After AI, cattle in the 10 million group had significantly lower pregnancy rates compared to those in the 18 and 20 million groups. Conversely, there were no statistically significant variances observed between cattle in the 10 and 15 million groups. Conclusions: Sperm at concentrations of 10, 15, 18 and 20 million per straw exhibited comparable motility, viability, and acrosomal membrane integrity. However, a concentration of at least 18 million sperm per straw is required to achieve a consistent rate of pregnancy rate in Hanwoo cattle after AI.

This study aimed to investigate the effects of amino acid complex additives, such as protected vitamin C (VC) or detoxified sulfur (DS), on the growth and metabolism of Hanwoo cattle under high-temperature conditions. Accordingly, farms in Temperature-Humidity Index (THI) regions ranging from 78 to 89 for over 100 days were selected. The experimental groups were control, T1 (lysine + methionine + VC, 50 g/head/day), and T2 (lysine + methionine + DS, 50 g/head/day) with 70, 77, and 71 animals each. The range of the THI for 115 days was 78-89, and this occurred in most of the experiment days. The results showed that there was no significant difference in rectal temperature among the groups. The body weight increased to 786.4 and 809.0kg in the T1 and T2 groups, respectively, compared to the control group (p<0.05). Linoleic acid showed a high result of 2.01% in the T1 group compared to the control group (p<0.05). Unsaturated fatty acids were higher at 55.70 and 56.54% in the T1 and T2 groups, respectively, compared to the control group (p<0.05), and the omega 6/3 ratio was reduced to 20.10% (p<0.05). These findings indicate that T1 has a positive impact on growth, meat quality, and fatty acid composition compared to the control group. In conclusion, amino acid complex with VC improved the body weight of Hanwoo steers and the unsaturated fatty acids and essential amino acids of their meat; however, further research is needed to clarify this impact on carcass performance.

본 연구에서는 거세방법에 따른 한우의 안심살(M. psoas major), 꽃등심살(M. longissimus thoracis) 및 우둔살(M. semimembranous)의 육질특 성과 지방산 조성을 조사하였다. 공시시료는 송아지가 7개월령에 도달하면, 외과적 수술방법을 통해 한쪽 고환을 제거한 반거세 한우(half-castration; HC) 9두와 양쪽 모두 제거한 완전거세 한우(complete-castration; CC) 8두를 이용하였다. 거세된 송아지는 24-26개월까지 동일한 조건에서 사육한 후 도축하고 3개 부위(안심살, 꽃등심살, 우둔살)를 채취하여 육질 분석하였다. 거세방법에 따른 그룹 간 지방함량은 HC 그룹이 2.97~11.06%로 CC 그룹보다 낮은 함량을 나타내었으며, 수분함량은 HC 그룹이 65.39~70.90%로 CC 그룹보다 높은 함량을 나타내었다(p<0.05). 3개 근육의 pH는 5.47~5.54 범위였으며, pH와 보수력은 거세방법에 따른 유의적인 차이는 나타나지 않았다(p>0.05). 육색은 L*값과 a*값에서 HC가 CC보다 낮게 나타났다(p<0.05). 가열감량과 전단력은 HC 그룹의 꽃등심살이 CC 그룹보다 높은 값을 나타내었다(p<0.05). 지방산 조성은 palmitic acid, palmitoleic acid, vaccenic acid, oleic acid, linoleic acid, linoleneic acid와 PUFA, n6에서 HC와 CC 그룹 간 유의적 차이가 나타났다(p<0.05). 특히, 지방산 조성에서 HC 그룹과 CC 그룹 모두 oleic acid가 가장 높은 함량을 나타내었다(p<0.05). 따라서 거세 방법은 한우육의 근육 내 지방, 육색, 연도와 같은 이화학적 특성과 지방산 조성에 영향을 미칠 수 있다고 사료된다.

본 연구는 한우 번식우에 있어서 영양대사물질 분석을 통하여 영양수준을 구명하여 번식우의 수태율 개선을 위한 기초 자료로 활용하기 위해서 실시하였다. 번식우의 정확한 영양수준 분석을 위해서 사료급여량을 80%, 100%, 120%로 구분하여 사양관리를 실시한 결과 Cholesterol과 BUN 농도가 120% 급여구에서 유의적으로 높은 결과를 나타내었다(p<0.05). 방목우 중에서 임신우와 비임신우의 영양대사물질 수준 분석 결과, Cholesterol, AST, NEFA 농도가 임신우에 비해 비임신우에서 유의적으로 높은 결과를 보였다(p<0.05). 이와 같이 임신과 관련한 영양수준 분석에 Cholesterol, AST, NEFA 의 3가지 항목을 설정하는 것이 필요할 것으로 사료된다. 방목과 사사 사육에 대한 결과 분석에서 Glucose 농도는 방목우 84.8, 비방목우 56.0 mg/dl 으로서 방목우에서 유의적으로 높은 결과를 보였고(p<0.05), Cholesterol 수준은 방목우에서 142.5 mg/dl로서 사사 사육 128.9 mg/dl 보다 유의적으로 높았으며(p<0.05), ALT(34.4 vs 27.1 IU/l)와 NEFA 농도(317.8 vs 160.2 ЧEq/l) 역시 방목우에서 유의적으로 높은 결과를 보였다(p<0.05). 결론적으로, 암소에 사료 급여시 Cholesterol, ALT, NEFA 수준을 낮출 수 있도록 하는 것이 한우 암소의 수태율을 높일 수 있을 것으로 사료된다.

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 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

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.

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.