This study was conducted to investigate the optimal artificial insemination (AI) time with diagnostic kit at ovulation time. We already applied the patent about the protein in the cow heat mucose in external reproductive tract. And we would examine the accuracy for detection of cow heat by the kit produced with the protein. Evaluation of optimal heat detection was tried two time at 12 hrs and 24 hrs after the heat. And then, AI service also performed two times with no relation to the results of heat diagnosis by heat detection kit and pregnancy rates were checked with rectal palpation on 60th day after AI. Heat diagnostic results by kit in natural heat after 12 hrs in Hanwoo cows were showed 31.3~75.0% on positive in first heat detection and 33.3~100.0% on positve in second heat detection. In the 1st positive results were significant different (p<0.05), but 2nd positive were not. The results of heat detection showed different result on regional influence and individual cow effects. The pregnancy rates of first trial of heat detection were showed 34.4~78.7% on positive and 21.3~68.8% on negative after the diagnosis by heat detection kit. And the pregnancy rates of next trial of heat detection were showed 33.3~85.7% on positive and 14.3~66.6% on negative after the heat diagnosis. Both positive results of first trial and next trial also were showed significant different (p<0.05), but negative results were not. In positive result, first trial of total pregnancy rates was higher than the next trial of pregnancy, but there showed opposite results on negative results. In conclusion, the optimal heat detection kit is suitable to ordinary Hanwoo cows and it suggested that we have to improve the kit’s accuracy by detecting the materials like proteins related optimal AI time.

축산농가에서 사육되는 주요 가축으로 한우, 젖소, 돼지, 가금류, 흑염소, 사슴, 말 등으로 이들 가축 들은 생산성을 높이기 위하여는 과학적인 번식관리를 통한 증식과 개량이 꾸준히 적용되어지고 있다. 농촌진흥청에서 축산농가에 제공되는 연구개발된 번식분야 실용화 기술들은 매년 보급되고 있으며 활 용도를 높이기 위하여 책자, DVD, 인터넷 등 매체를 통하여 보급되고 있다. 이와 같이 1993~2016년 동안 제공된 영농활용기술 중 번식분야를 중심으로 제공된 기술을 분석하여 향후 축산농가 번식기술 방향을 제시하고자 조사를 실시하였다. 1993~2016년 동안 번식분야 영농활용기술은 총 407건으로 축 종별로는 한우 170건, 젖소 94건, 돼지 97건, 가금류 18건, 흑염소 12건, 사슴 8건, 말 7건 등이었고, 번식 기능별로는 번식관리 167건, 인공수정 99건, 수정란이식 45건, 교배계획 31건, 가금번식 14건 등 순이었 다. 번식관리 분야는 전체 167건에서 호르몬 처리에 의한 번식력 향상 기술이 35건, 사료 사양관리 17, 비타민급여 번식효율 향상 9, 신체충실지수(BCS) 7건, 발정관찰 5건, 임신진단 4건 등 순이었다. 인공 수정 분야는 전체 99건에서 인공수정 일반기술이 42건, 정액제조 21건, 정액선택 16건, 정액보존 11건, 정액채취 6건 등 이었다. 수정란이식 분야는 전체 45건에서 호르몬처리에 의한 수정란이식 일반관리기 술 16건, 다배란유기 기술이 12건, 신체충실지수 적용기술이 4건, 수정란 성판별 기술 2건 등이었다. 가금류 번식분야는 14건에서 부화기술 5건, 종계관리 4건, 종란관리 3건, 인공수정 2건이었다.

A total of 30 Korean native pigs (gilt 15, boar 15) were used to investigate the carcass characteristics, meat quality, amino acid, and fatty acid composition by gender. The carcass weight of boars were significantly higher than gilts, whereas the carcass yield of gilts had significantly higher than boars (p<0.01). Boars had significantly higher moisture contents in loin muscle than gilts, whereas the protein contents of loin muscle had significantly higher in gilts than boars (p<0.01). In the results of meat quality analysis, the cooking loss (p<0.01), shearing force (p<0.05), lightness (L) and yellowness (b) in meat color (p<0.05) were significantly higher, but the pH was significantly lower (p<0.01) in gilts compared with boars. Arginine (p<0.05), alanine, aspartic acid, histidine, leucine, lysine, phenylalanine, serine, threonine and tyrosin (p<0.01) for gilts were significantly higher than those for boars. The results of fatty acid composition showed that gilts had significantly higher contents of C16:1n7, C18:1n9, C20:1n9 (p<0.01) than boars in intramuscular fat, whereas boars had significantly higher contents of C18:2n6, C20:4n6 (p<0.01) and C18:3n3 (p<0.05) than gilts in intramuscular fat.

This study was designed to determine whether low-density lipoporoteins (LDL) extracted from egg yolk in extender improve the function of Korean Jeju Black Bull semen. The semen was cryopreserved with 5% ethylene glycol (EG) or 7% glycerol (G) extenders containing 10% egg yolk (EY), 4% LDL and 5% EY or 8% LDL. Frozen-thawed sperm were evaluated sperm motility, viability, membrane integrity and acrosome integrity. Post-thawed sperm motility has been significantly higher (p<0.05) in 4% LDL + 5% EY (; EG and ; 7% G) than 8% LDL (; EG and ;G). Treatment of 4% LDL + 5% EY-EG () has been significantly improved sperm viability compared to other treatments except 10% EY - EG. Moreover, in membrane integrity, swollen sperm ratio has been only significantly increased (p<0.05) in 4% LDL + 5% EY - EG () among all treatments. In assess to detect acrosome integrity, especially, AR pattern ratio has been significantly decreased (p<0.05) in 4% LDL + 5% EY - EG among all treatments. In sperm viability as time passes, between 4% LDL + 5% EY and 10% EY, there was no significant difference, but 8% LDL was significantly decreased sperm viability in EG (1 and 2 hrs) and G (30 min, 1, 2, 5 and 12 hrs) extender. However, there were no significant differences among all treatments except 8% LDL-G in sperm membrane integrity. 8% LDL-G has been significantly decreased swollen sperm ratio at 5 hrs after thawed. It is concluded from these results that 4% LDL + 5% EY to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Korean Jeju Black bull.

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 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).

포유동물 수정란의 동결보존기술은 최근 기후 변화에 따른 생물종 다양성을 보존하기 위해서 중요하게 여겨지는 연구 분야이다. 따라서 멸실 위험에 처한 동물의 개량과 증식, 보존과 복원 및 생명공학의 분야에 이르기까지 응용 기술은 다양하게 이용되어진다. 본 연구에서는 한우 수정란의 동결 후 생존성 향상을 위해서 동결 방법에 따른 체내 외수정한의 내동성을 조사하였다. 완만동결에 따른 체내 외수정란의 동결 융해 후 수정란의 재확장률은 89.6%와 81.5% 그리

This study was carried out to establish most suitable freezing condition, to evaluate the different glycerol concentration of freezing and thawing rates on motility, viability, membrane integrity and acrosome intecrity of frozen Korean Jeju Black Bull spermatozoa, Semen was collected from a Korean Jeju Black Bull using an artificial vagina and transported to the laboratory. The semen was extended gradually 1:5 then cooled slowly for 2 hrs to 4. The semen was diluted 1:1 with cryoprotectant extenders (3%, 5% and 7% glycerol) and equilibrated for 2 hrs at cold chamber and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 min and above 8 cm for 10 min. And then the frozen straw was plunged into LN. The presented straws were examined the viability and motility after thawed at 37 water bath. The viability and membrane integrity immediately post-thawing were significantly higher in samples frozen in 7% glycerol than 3% and 5% glycerol (p<0.05). After CTC staining to assess acrosome integrity, F pattern was significantly increased, but B pattern was significantly decreased in 7% glycerol (p<0.05). Freezing distance of 5 cm from liquid nitrogen and pre-cooling for 10 min yield better survival and membrane integrity, but not significant difference. However, AR pattern according to CTC staining was significantly decreased in 3 cm for 5 min.

Previously, we reported that the osmolarity conditions in the satellite region were affected CpG DNA methylation status while Pre-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. This study was conducted to investigate the DNA methylation status of repeat sequences in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaCl or 0.05 M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. The DNA methylation status of the Pre-1 sequences in blastocysts was characterized using a bisulfite-sequencing method. Intriguingly, in the present study, we found the unique DNA methylation at several non-CpG sequences at the Pre-1 sequences in all groups. The non-CpG methylation was hypermethylated in all three groups, including in vivo group (86.90% of PZM- 3; 83.87% of NaCl; 84.82% of sucrose; 90.94% of in vivo embryos). To determine whether certain non-CpG methylated sites were preferentially methylated, we also investigated the methylation degree of CpA, CpT and CpC. Excepting in vivo group, preference of methylation was CpT>CpC>CpA in all three groups investigated. These results indicate that DNA methylation of Pre-1 sequences was hypermethylated in CpG as well as non-CpG site, regardless modification of osmolarity in a culture media.

This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.

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

The objective of this work was to determine the effect of corpus luteum (CL) grade on pregnancy rate after embryo transfer in Korean cattle and we found that CL development was linked to pregnancy rate. The in vivo derived blastocyst-stage embryos were transferred to 15 recipients synchronized in the estrus cycles. Based on size and palpable characteristics, CLs were categorized into three grade. The grade three CL is not to be identified by rectal palpation. The pregnancy rates tended to increase with the increase in CL size of recipients. In grade one, two, and three, the pregnancy rates were 62.5%, 50.0%, and 0%, respectively. This result suggests that pregnancy rates after embryo transfer might be affected by the CL status of recipients.