칡소의 귀세포를 이용한 핵이식 시에 전기 융합조건이 핵이식란의 발생에 미치는 영향을 알아보고, 칡소의 귀세포와 한우의 태아섬유아세포를 이용한 핵이식란을 이식하여 수태율을 비교하였다. 전기자극 조건에 따른 핵이식 후 융합율은 51∼68%의 범위로 차이는 없었으나, lysis 율은 1.9kv/cm의 10us, 20us에서 각각 0.0과 38.7%, 2.0kv/cm의 10us, 20us에서 각각 2.9와 37.5%, 2.1kv/cm의 10us, 20us에서 각각 21.2와 51.8%을 보였다. 난할율은 1.9kv/cm의 10us, 20us에서 각각 75.8과 69.8%, 2.0kv/cm의 10us, 20us에서 각각 76.9와 68.8%, 2.1kv/cm의 10us, 20us에서 각각 70.5와 68.5로 큰 차이가 없었다. 그러나 배반포 발생율은 1.9kv/cm의 10us, 20us 각각 19.5와 48.6%, 2.0kv/cm의 10us, 20us에서 각각 20.0과 40.9%, 2.1kv/cm의 10us, 20us에서 각각 44.2와 27.0%로 각 조건에서 시간에 따른 차이를 보였다. 핵이식에서 공여세포와 세포질간의 융합율과 lysis율은 전압과 통전시간에 영향을 받으며 전압을 높이는 것이 통전시간을 길게 하는 경우보다 수핵난자의 세포질에 상해를 줄이고 배반포 발생에 유리 하였다. 한우의 태아섬유아세포의 핵이식후 생산된 배반포를 5마리의 수란우에 이식한 결과 두 마리가 임신되었고(40.0%), 칡소의 귀 세포를 핵이식한 후 생산된 배반포를 19마리에 이식한 결과 3마리가 임신되었다(15.8%).

The ultrasound-guided oocytes cllection (ovum pick-up ; OPU) has become a substitution for superovlation in cattle. The objective of this study was to examine the effect of OPU frequency on the in vitro production of embryos in Hanwoo cattle. Six cycling Hanwoo cows were distributed into two groups for either once or twice weekly OPU sessions. Oocytes were collected by ultrasound-guided follicle aspiration(SA600) using a 6.5HMz transducer and attached with 18 gauge needle, with vacuum pressure of 40 mmHg. The cumulus-oocyte complexes (COCs) collected from each donor were matured in TCM 199 supplemented with 10% fetal bovine serum at 5% CO2 in air at 38.5 for 22h and in vitro matured oocytes were co-incubated with sperm(separated by Percoll gradient) for 6h. The zygotes were co-cultured on cumulus cell monolayer in 10ul droplets in the same culture medium and conditions used for IVM for 7 days. On Day 7 of culture, development to blastocysts was examined. Although the number of oocytes collected was variable depending on individuals, overall embryo production in the twice per week OPU sessions was better that in the once per week sessions(6~21 vs 2~7 blastocysts produced, respectively). Two cows(E, A) were good oocyte donors and embryo production was superior in cow C ; however, cow F was a poor donor as compared to the others. In conclusion, these results suggest that for embryo production, twice weekly OPU sessions were better than once per week for producing embryos in vitro from Hanwoo cattle

This study was carried out to investigate the effect of co-culture system(bovine oviduct epithelial cells; BOEC) and defined culture system (modified TALP ; mTALP) on the conception of embryos transferred, and pregnancy and twin birth rates after transfer of fresh or frozen-thawed bovine blastocysts produced in vitro were also evaluated. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol. The results obtained were as the following. The pregnancy rate after transfer was higher in co-culture group than in mTALP group, but was not signficantly different, and there is no difference between fresh embryo group and frozen-thawed embryo group in conception rate. The conception rate was not different whether 3∼4 blastocysts or 2 blatocysts transferred into a recipient, but the production rate of twin calves was significantly higher (p<0.05) when 3∼4 embryos transferred. The average birth weight of twin calves(24.38kg) was numerically, but not significantly lighter than that of single calves(26.68kg).

This study was carried out to increase the viability of bovine frozen4hawed in vitro produced (IVP) embryos and pregnancy rate by direct transfer method. Cumulus-oocyte complexes were aspirated from excised Hanwoo ovaries and matured in TGM 199 for 20~22 hours at 38.5 in 2% in air. Matured oocytes were fertilized with capacitated sperm for 6 hours and then co-cultured with cumulus cells for 9 days. 63% of the oocytes cultured was deaved and 29% out of them developed into blastocysts. Good or excellent grade of blastocysts on D 7 or 8 were frozen with 1.8M ethylene glycol as a cryoprotectant for direct transfer. Frozen embryos were thawed at 2 water for 10 sec following 4~5 second in air. For the survival assay of frozen4hawed lVP blastocysts, they were cultured in TCM 199 supplemented with 100M -mercaptoethanol and 20% FCS for 72 hours. The percentage of embryos developed to re-expanded or hatched after 72 hours culture was 95. 5 and 77.3%, respectively. When frozen-thawed Ivp embryos were transferred to 43 synchronized recipients by direct transfer method, eighteen recipients (41.8%) was pregnant. The highest pregnant was in naturafly synchronized recipients (71.4%), but induced estrus by using PRID(29.2%) and PGF(20.0%) was showed lower pregnancy rate. The pregnancy rate was higher in day 7 blastocysts(56.0%) than day 8 blastocysts(22.2%). (Key words: in vitro produced, blastocyst, frozen-thawed, direct transfer)

This study was aimed to grow organic forage as a prime requirement for organic Hanwoo cattle production over a period of 3 years (2012~2015) in Gyeongbuk region, Korea through mixed cropping of winter fodder barley with legume and application of fermented cattle manure. The effect of barley-legume mixed cropping and application of different levels of fermented cattle manure was studied on forage productivity of whole-crop barley (WCB) and evaluation of feeding capacity of Korean native cattle (Hanwoo) per unit area. The collected data was analyzed using SAS 9.3 software. Barley-legume mixed cropping increased (P< 0.05) yields of dry matter (DM), crude protein (CP) and total digestible nutrients (TDN) as compared to those of sole WCB as monoculture. Similarly, acid detergent fiber (ADF) contents and relative feed value (RFV) were also higher (P<0.05) in barley-legume mixed sowing forage compared with those of sole WCB as monoculture. In case of manure application, yields of DM, CP and TDN were also found (P<0.05) highest in 100~150 ㎏ N/㏊. However, varying cattle manure levels did not influence (P>0.05) CP and neutral detergent fiber (NDF) contents. In response to increasing nitrogen application level, ADF content increased (P<0.05) but TDN content and RFV decreased (P<0.05). The Barley-legume mixed sowing and increasing application levels of fermented cattle manure also significantly enhanced (P<0.05) feeding capacity of organic Hanwoo (head/ha) having 450 kg body weight with 400 g of daily gain. Findings of present study indicated that barley-legume biculture not only cut nitrogen application level of fermented cattle manure by over 50 kg per ha but also increased their feed value and productivity of forage as compared to WCB monoculture. This would be contribution to the diversity of forage resources as well as production of organic animal product by creating low cost and high quality organic forage.

This study was conducted to investigate the effects of mixed-sowing of legume forage and applying of cattle manure on the productivity of organic rye to provide dairy farmers with safe organic feeds. Also the present study aimed to evaluate optimal applying level of cattle manure and carrying capacity of Korean native cattle (Hanwoo) per unit area. The amount of crude protein was significantly higher in the mixed-sowing of legume forage (average 760~786 kg/ha) than in the single-sowing of rye (average 353 kg/ha) (p<0.05). The mixed-sowing of forage peas and the single-sowing of rye tended to improve by increasing the level of nitrous fertilization rather than the fertilization treatment. The amount of crude protein on the rye according to the mixed-sowing of legume was significantly higher in the mixture (average 8.29~9.90%) than in the single (average 4.93%) (p<0.05). The comparison by the level of nitrogen fertilization indicated signifi-cantly high for the rye in 50 kg N/ha than in fertilization treatments (p<0.05). Total digestible nutrients (TDN) amount in the single-sowing of rye was average 46.86% and has lower than average 49.96~50.12% of the mixed-sowing of legume, and especially the mixed-sowing of forage pea was the highest with 54.55% in 150 kg N/ha for the level of nitrogen fertilization (p<0.05). The breeding ability of annual organic livestocks per unit area according to the feed value of rye presented significantly higher ability in the mixed-sowing of legume (3.72~4.12 heads) than the single-sowing of rye (average 2.26 heads) (p<0.05). By summarizing above results, the mixed-sowing of legume is required to improve the productivity and the feed value of rye for increasing organic livestock breeding ability in southern area of Gyeongbuk regions, and the study for identifying the appropriate fertiliza-tion level using livestock excretions is further necessary.

본 연구는 토양오염을 감소시키고 유기축산물의 생산에 기여할 수 있는 유기조사료를 효율적으로 생산하기 위해 대표적인 동계작물인 총체 밀에 콩과 사료작물의 혼파와 발효우분을 시용하여 생산성과 사료가치를 평가하고, 토양의 지력 향상과 가축분뇨의 자원화를 위한 단위면적당 유기한우 사육능력을 추정하고자 하였다. 콩과 사료작물의 혼파에 따른 2년(2012~2013년) 평균 총체 밀의 건물, 조단백질 및 TDN 수량은 6.10~7.70, 0.59~0.76 및 3.84~4.74톤/ha으로 단파로 재배했을 경우의 4.07, 0.35 및 2.54 톤/ha 보다 유의하게 증가하였으나 혼파구 간에는 차이가 없었고, 발효우분의 질소시용수준이 증가함에 따라 총체 밀의 평균 건물, 조단백질 및 TDN 수량은 100~150 kg N/ha(7.24~7.59, 0.67~0.73 및 4.43~4.69톤/ha)가 무비구(5.96, 0.57 및 3.71톤/ha)에 보다 유의하게 증가하였다(p<0.05). 총체 밀의 조단백질 함량은 콩과사료작물의 혼파구에서 평균 9.83~10.03%로 총체 밀 단파구의 8.37%보다 유의하게 높았고 ADF 함량은 헤어리 베치 혼파구 그리고 RFV는 사료용 완두 혼파구가 다른 처리구 보다 유의하게 높았으며(p<0.05), 발효우분의 질소시용수준에 따른 조단백질 함량은 9.17~9.52%로 처리수준 간 차이가 없었고 ADF 함량은 시용수준이 100 kg N/ha 까지 증가함에 따라 유의하게 증가하였고 TDN 함량은 반대로 유의하게 감소하였다(p<0.05). 콩과 사료작물의 혼파에 따른 총체 밀에 의한 유기한우 사육능력(체중 450 kg, 400 g 일당증체 목표)은 평균 3.99~4.20두로 단파구(평균 2.21두) 보다 유의하게 증가하였고. 발효우분 시용수준이 증가됨에 따라 질소 수준 100 kg/ha(평균 3.68두)까지 무비구(평균 3.09두) 보다 총체 밀에 의한 유기한우 사육능력도 유의하게 증가하였다(p<0.05). 이상의 결과를 종합해 볼 때, 총체 밀 재배 시 콩과 사료작물의 혼파 하였을 경우에는 단파재배 보다 발효우분의 질소시용수준을 ha 당 50 kg 이상 줄이면서도 건물수량을 포함한 생산성과 유기한우 사육능력 증대는 물론 조단백질 함량 등 사료가치를 향상을 시켜 한우사양 시 조사료자원의 다양화 뿐 아니라 저투입에 의한 양질의 조사료 생산으로 유기축산물 생산에 크게 기여하리라 사료된다.

본 연구는 환경오염을 감소시키고, 안전한 유기축산물의 생산에 기여할 수 있는 유기조 사료를 효율적으로 생산하기 위해 대표적인 동계작물인 호밀에 콩과 사료작물 혼파와 발효우분을 시용하여 생산성과 사료가치를 평가하고, 토양의 지력 향상과 가축분뇨의 자원화를 위한 가축분뇨의 적정 시용수준을 구명하여 단위면적당 유기 한우의 사육 능력을 추정하고자 하였다. 호밀의 연간 조단백질 수량은 지역 시험구 비교에서는 경주 시험구가 경산과 영주 시험구 보다 높게 나타났으며, 시험구별 단·혼파 비교에서는 사료용 완두 혼파구가 가장 높게 나타났다(p<0.05). 발효우분 시용수준에 따른 조단백질 수량은 모든 시험구에서 시용수준이 높을수록 증가하였고 ha당 100, 150kg 수준에서 유의하게 높았다(p<0.05). 사료작물의 사료가치 평가에서 조단백질 함량은 콩과작물의 월동이 가능한 시험구에서는 호밀 단파보다는 콩과 혼파구가 높은 경향으로 나타났고, 특히 콩과 작물 중 사료용 완두가 높게 나타났다(p<0.05). 유기 한우 사육능력은 경주 시험구가 ha당 3.28두로 경산 시험구의 ha당 1.94두, 영주 시험구의 ha당 1.50두 보다 매우 높았으며(p<0.05), 호밀 단파 보다는 콩과 작물혼파구가, 혼파구에서는 사료용 완두가 높게 나타났다(p<0.05). 이상의 결과를 종합해 볼 때, 사료가치 증진을 위한 콩과 사료작물 혼파는 겨울철 콩과작물의 월동성을 고려해야 하며, 콩과작물의 월동이 어려운 지역에는 발효우분을 시용하는 것이 유리하고, 또한 단위면적당 수량과 단백질 함량 등의 사료가치를 높이기 위해서는 발효우분 수준을 ha당 100~150kg 시용과 함께 콩과작물은 사료용 완두를 혼파재배하는 것이 유기 가축사육 능력 증대를 기대할 수 있으리라 사료되었다.

The objective of this study was to evaluate the effect of applying alum (aluminum sulfate) and aluminum chloride on pH and pathogen populations of Hanwoo manure. A total of 36 steers (8 months old and averaging 300 kg in weight) were used in this trial and allotted to 9 pens (3 replication pens per group with 4 steers per experimental unit, 5 x 8 m). Chemical additives were applied as a top dressing with garden rake to a depth of 1 cm of manure with wood shavings in each treatment. The chemical amendments were control (without chemical amendments), 50 g of alum and 50 g of aluminum chloride/kg of Hanwoo manure. The experiment was carried out for 4 weeks. Adding alum and aluminum chloride to Hanwoo manure reduced (P < 0.05) pH compared to untreated controls during the 4-wk period. Both levels of the alum and aluminum chloride treatments tested decreased (P < 0.05) Escherichia coli and Salmonella enterica populations in Hanwoo manure at 2 and 4 weeks. It appears that the reduction in pathogen populations was primarily associated with the lower manure pH. If more strict environmental regulations are put into effect regarding pathogen populations from Hanwoo facilities, treating Hanwoo manure with alum and aluminum chloride may be a good management practice.