This study was conducted from October 2014 to May 2015 to explore forage production and feed values of Italian ryegrass, Rye and whole crop barley as winter forage crops in the Southern region of Korea. The experimental location was over 10 points for each species and each sampling point area was 1 m² (Width: 1 m × Length: 1 m). Air mean temperature and rainfall in the Southern region of Korea during the experimental period was 6.95 ± 5.75℃ and 70.45 ± 54.68 mm, respectively. Fresh forage yield of Italian ryegrass, the most cultivated forage in the Southern region of Korea, was 44.4 ± 7.0 ton/ha. The percentage of dry matter for whole crop barley was 28.9 ± 7.0%. Crude protein (CP) was higher in Italian ryegrass (10.7 ± 5.3%) while total digestible nutrient (TDN) had the highest value in whole crop barley. Crude protein was not significantly different by location. However, the neutral detergent fiber (NDF), acid detergent fiber (ADF) and total digestible nutrient value of forage from Jeonbuk province were higher than in forage from Gyeongnam province.

본 연구는 국내에서 헤일리지의 생산 및 이용 확대를 위해 수분함량에 따른 헤일리지의 사료가치 및 발효품질의 변화를 구명하고 양질의 헤일리지 생산 확대를 위한 기초 자료를 확보하기위하여 2011년 9월부터 2012년 12월까지 충남 천안시 소재 국립축산과학원 초지사료과 시험포장에서 수행되었다. 수분함량이 다양한 헤일리지의 조제를 위해 이탈리안 라이그라스의 ‘코윈어리’ 품종을 이용하여 수분함량 60~30%까지 5% 차이로 총 7처리로 헤일리지를 조제하였다. 수분함량이 다른 이탈리안 라이그라스 헤일리지의 사료가치는 수분함량에 따라 유의적인 차이를 나타내지 않았다. 수분함량에 따른 발효품질의 변화는 수분함량의 차이에 따라 유의적인 차이를 보였으며 상대적으로 수분함량이 높은 조건(>40%)에서 발효품질이 우수하였으며 수분함량이 낮은 조건에서는 발효품질도 낮은 것으로 나타났다. 수분함량별 헤일리지의 유기산 구성비율은 수분함량이 상대적으로 높은 조건(>40%)에서 젖산비율이 높게 나타났으며 수분함량이 낮은 조건(<40%)에서는 젖산비율이 감소하고 초산과 낙산비율이 높아지는 것으로 나타났다. 이상의 결과를 종합해 보면 안정적인 헤일리지 생산을 위해 발효품질을 고려해보면 수분함량이 40%가 헤일리지 조제의 한계점으로 사료되며 추후 헤일리지의 장기 저장성 검토 및 저 수분 조건에서 발효품질 개선을 위한 추가적인 연구가 진행되어야 할 것으로 생각된다.

본 연구는 근적외선분광법을 이용하여 우리나라에서 재배되고 있는 목초류 중 외형적 특성이 유사한 이탈리안 라이그라스, 페레니얼 라이그라스와 톨 페스큐 종자의 초종 판별 가능성을 검토하고자 수행되었다. 근적외선분광기를 이용하여 목초류 종자를 가시파장 대역대(680~1,099 nm), NIRS 파장 대역대(1,100-2,500 nm) 및 NIRS 전체 파장 대역대(680-2,500 nm)로 구분하여 스펙트라를 얻은 후 1차 미분과 8 nm gap으로 수 처리를 수행하였으며 부분최소자승(PLS) 회귀분석법을 통해 초종판별 검량식을 개발하고 판별 정확성을 검증하였다. 목초류의 초종판별 정확성은 가시파장대역에서 SECV 1.732, R2CV 0.96으로 가장 판별 정확성이 낮았으며 NIRS 전체 파장대역에서 SECV 1.182, R2CV 0.98로 가장 높은 판별 정확성을 나타내었다. 파장대역별 예측 정확성은 NIR 파장대역(1,100-2,500 nm)에서 교차검증오차(SECV) 1.319에서 예측 오차(SEP) 1.288로 낮아졌으며 가시영역대(680~1,099)는 SECV 1.732에서 SEP 1.749로 약간 높아졌다. Discrimination equation 분석법에 의한 NIRS 전체 파장대역별 목초류 초종의 판별 결과는 초종간에 판별 정확성의 차이가 크게 나타났으며 이탈리안 라이그라스의 ‘Hits’는 68%로 가장 낮았으며 페레니얼 라이그라스가 78%의 정확성으로 가장 높게 나타났다. 따라서 NIRS를 이용한 목초류 초종의 판별분석이 가능할 것으로 판단되었다.

본 연구는 감꽃의 주요 향기성분을 알아보기 위해서 감나무 수령별로 감꽃의 향기성분을 SPME를 이용하여 분석하고, 기관을 꽃과 꽃받침으로 분류하여 향기성분을 비교하였다. 감나무 수령별 감꽃의 만개율은 15년생 이상, 10-14년생, 5-9년생 순으로 수령이 오래될수록 초기 만개율이 높았다. 감나무 수령에 따른 감꽃 향기성분 분석 시 동정된 주요 향기 성분은 a-pinene, butene, caryophyllene, cubebene, lavandulol, D-limoneneylangene, ylangene 등의 성분을 얻을 수 있었다. 대부분이 green 향, fruit계, floral계의 옅은 향도 포함하고 있었다. 감꽃에 존재하는 휘발성 향기성분의 수는 5-9년생은 30종, 10-14년생은 24종, 그리고 15년생 이상에서는 32종으로 수령이 5년 미만과 15년 이상에서 많은 향기성분이 조사되었다. 감꽃을 꽃받침과 나누어 향기성분을 비교하면 단감 ‘부유’ 품종 중 꽃의 향기 성분은 10개이고 상대적인 총 함량은 26.35%이며, 꽃받침은 향기 성분은 14개 이고 상대적인 총 함량은 46.28%로 꽃에 비해서 더 다양한 향기성분이 존재하는 것으로 조사되었다. 반면 떫 은감 ‘둥시’ 품종은 꽃에서는 6개의 향기성분이 17.58%, 꽃받침에서는 9개의 향기 성분이 50.27%로, ‘부유’ 품종 에 좀더 다양한 향기성분이 존재하였다. 본 연구는 감꽃 향기를 이용하고자 하는 향기산업에 기초자료를 제공하 는데 기여할 것이다.

In this study, we aimed to examine the cellular and molecular mechanisms of KTH-13 (4-isopropyl-2,6-bis(1-phenylethyl)phenol) which is derived from Cordyceps staphylindaecola in the cancer cells survival. The apoptotic effect of KTH-13 on various cancer cells, such as C6 glioma, MDA-MB-231 breast cancer, and A549 cells, was determined by MTT assay, and result showed that KTH-13 (0-100mM) dramatically inhibited the cancer cell survival. IC50 of KTH13 were 60.549, 53.512, >100, in C6, MDA-MB-231, and A549, respectively. DNA fragmentation result revealed that MDA-MB-231 cells treated with KTH-13 100mM undergoes apoptosis. To understand the action mechanism of KTH-13, the effect of KTH-13 on caspase which is key regulator of apoptosis was verified. The amount of cleaved capspase-3 and 7, executioner caspases, was increased by KTH-13 treatment, at time dependent (capspase-3 case) and dose dependent manner (caspase-7 case). And the cleavage of caspase-9 which is initiator caspase was also elevated in KTH-13 treated MDA-MB-231 cells showing time dependent manner. However, caspase-8 was not regulated by KTH-13, indicating KTH-13 specifically targets caspase-9 signal. As caspase-9 is closely associated with intrinsic pathway, the involvement of bcl-2 family was identified. Bax, pro-apoptotic molecule, was up-regulated whereas Bcl-2, anti-apoptotic protein, was down-regulated. And the Bax/Bcl-2 ratio was increased about 10 times. Then, the survival signal was also observed. The phosphorylation of Akt and p85 was diminished by KTH-13 treatment at 2,4,6 and 8 hour. Collectively, results suggest that KTH-13 induces cancer cells apoptosis via caspase3, 7, 8 and Bcl-2 family signaling pathway. And the Akt and p85 is also involved in KTH-13 action mechanism.

본 연구에서는 이탈리안 라이그라스와 헤어리 베치를 이용하여 사일리지 제조시 젖산균과 클로렐라를 처리하여 사일리지의 사료가치, 품질 및 미생물상의 변화를 조사하였다. IRG-HV 사일리지 제조시 대조구와 젖산균 단독처리구의 조단백질 함량은 비슷한 수준을 보였으나 클로렐라와 젖산균 공용처리구는 현저하게 증가하였다(p<0.05). 섬유소와 TDN 함량 그리고 in vitro 건물소화율 처리구에서는 차이를 나타나지 않았다. IRG-HV 사일리지에서 대조구는 젖산균 처리구의 pH와 비슷하였으나 클로렐라와 젖산균 공용처리구에서는 대조구에 비해 pH가 감소되었다(p<0.05). 젖산함량은 대조구에 비해 젖산균 첨가구에서 증가되었으며(p<0.05), 클로렐라와 젖산균 공용처리구에서는 대조구에 비해 젖산함량이 현저하게 증가하였다(p<0.05). IRG-HV 사일리지 젖산균 단독 처리구의 젖산균 수는 대조구에 비해 현저하게 증가하였으며(p<0.05). 또한 클로렐라와 젖산균 공용처리구도 현저하게 증가하였다.이상의 결과를 요약해 보면 IRG-HV 사일리지 제조시 젖산균과 클로렐라 처리에 의해 사일리지의 품질이 향상되었다.

본 연구는 우리나라에 난지형 목초의 도입 가능성을 검토하고 난지형 목초의 재배 한계지를 구명하기 위하여 전남 장흥, 광주광역시, 전북 김제 및 충남 천안에서 2009년 5월부터 2012년 12월까지 수행하였다. 난지형 화본과 목초인 버뮤다그라스 “Common”, “자생형(Ecotype)” 품종, 바히아그라스 “Argentine”, “Tifton 9” 품종과 클라인그라스 “Selection 75” 품종을 2009년 5월 하순에 파종하였다. 버뮤다그라스의 출현 소요일은 파종 후 12일정도 소요되었으며 바히아그라스는 파종 후 24~28일 정도 소요되는 것으로 나타났다. 월동이 가능한 광주지역에서 버뮤다그라스의 출수기는 5월 28일경, 바이하그라스는 7월 26일경, 클라인그라스는 5월 30일경에 출수기에 도달하였다. 월동성은 전남 장흥 및 광주지역에서는 바히아그라스를 제외하고는 모두 월동이 가능하였으며 전북 김제와 충남 천안지역은 월동이 불가능한 것으로 나타났다. 난지형 목초의 건물생산성은 모든 시험지역에서 클라인그라스가 6,106~15,331kg/ha으로 가장 높게 나타났으며 바히아그라스가 가장 낮은 것으로 나타났다. 단백질 함량은 버뮤다그라스의 Ecotype이 12.78%로 가장 높게 나타났으며 ADF 함량은 버뮤다그라스 common이 30.42%로 가장 낮은 것으로 나타났다. 총 가소화영양소(TDN) 함량은 버뮤다그라스가 68.23%로 바히아그라스 보다 높은 것으로 나타났으며 in vitro 건물소화율은 초종간에 큰 차이를 보이지 않았으며 버뮤다그라스가 다소 높은 경향을 보였다. 이상의 결과를 종합해보면 우리나라에서 난지형 목초 활용을 위한 재배한계 지역은 시험기간 동안 지속적으로 월동이 가능한 전남지역까지로 판단되며 연간 사초생산성과 사료가치는 한지형 목초와 비슷한 생산성과 사료가치를 보였다. 하지만 기후 온난화 대응 측면과 한지형 목초의 여름철 수량감소로 인한 지속성 저하를 보완 대체할 수 있는 조사료 자원으로서 난지형 목초의 새로운 접근이 필요할 것으로 생각된다.

Rutin is one of the major flavonoids found in buckwheat (Fagopyrum esculentum Moench). While rutin is already known to exhibit anti-oxidative, anti-inflammatory, and anti-carcinogenic activities. However, the health beneficial function of rutin metabolites is not well understood. In DPPH radical scavenging assays, the present study found that 3,4-dihydroxyphenyl acetic acid had the highest total anti-oxidant activity, followed by 3,4-dihydroxyphenylacetic acid, rutin, homovanillic acid, and 3-hydroxyphenyl acetic acid. Further, 3,4-dihydroxyphenylacetic acid strongly reduced LPS-induced IL-6 production in RAW 264.7 cells, compared with other metabolites. Therefore, these results suggest that rutin metabolites have potential to be utilized as food ingredients with anti-oxidant and anti-inflammatory activities.

For more than two decades, the intracytoplasmic sperm injection (ICSI) technique has been used as a valuable tool to provide opportunities for studying fertilization, treating human infertility, and producing transgenic animals. Not only in facilitating fertilization but also in propagating mammalian species, ICSI has enhanced the potential of assisted reproductive technologies in human. Polyspermic fertilization has been one of major problems in pig reproduction, but the ICSI helped to solve the problem, and used widely to generate transgenic piglets. Although the ICSI technique is considered to be a very useful tool in assisted reproductive technologies, including generation of transgenic animals, there are some disadvantages using the technique. In this review, we describe the ICSI technique and its application in animal production and human infertility, and discuss advantage and disadvantage of the technique in mammals.

The objective of this study was to investigate the effects of NEAA and leptin supplemented to in vitro culture medium on the developmental competence of porcine embryos after intracytoplasmic sperm injection (ICSI), and to modify the culture condition to improve the quality and the development of ICSI-derived porcine embryos in vitro. After ICSI, the putative zygotes were then cultured in PZM-3 medium with/without NEAA or leptin. The proportion of embryos that developed to the blastocyst stage significantly increased when 1% NEAA (24.62%) was added to the medium compared with 2% NEAA and no NEAA group (17.24% and 20.24%, respectively, p<0.05). The effect of different concentration of leptin (0, 10, 100, 500 ng/ml) was evaluated on the development of porcine ICSI embryos cultured in vitro. In case of blastocyst formation, 100 ng/ml group (27.05%) showed significantly higher rate than 10, 500 ng/ml, and control group (23.45%, 17.99%, and 19.68%, respectively, p<0.05). We also evaluated the effects of different NEAA and leptin treatment time on the development of porcine embryos after ICSI. Among groups of embryos cultured in the presence of NEAA or leptin for whole 7 days (D 1-7), first 4 days (D 1-4), the subsequent 3 days (D 5-7), both NEAA (27.13%, 21.17 %, and 17.56%, respectively, p<0.05) and leptin (25.60%, 20.61%, and 16.53%, respectively, p<0.05) showed that supplementation for whole 7 days significantly increased the blastocyst formation rate compared with the other groups of D1-4 and D5-7. We further evaluated the combination effect of 1% NEAA and 100 ng/ml leptin compared with the effect of each supplementation with 1% NEAA or 100 ng/ml leptin or no supplementation on development of embryos. For blastocyst formation, combination group of NEAA and leptin (24.78%) showed significantly higher rate than other three groups (18.37%, 20.44 %, and 13.27%, respectively, p<0.05). We further evaluated the expression of proapoptosis genes such as BAX and BAK and anti-apoptosis genes, BCL-XL and BCL-2 in blastocysts cultured in the presence of 100 ng/ml leptin. RT-PCR analysis revealed that leptin supplementation significantly decreased the expression of pro-apoptosis genes as well as increased the expression of anti-apoptosis genes. These results of present study demonstrate that NEAA and leptin could improve the in vitro development of ICSI- derived porcine embryos with optimal concentration of each reagent. Furthermore, the optimal culture condition could increase the quality of ICSI-derived embryos in vitro.

True hermaphrodites are animals of equivocal sex in which both male and female gonads develop simultaneously. The frequency of true hermaphroditism is higher in pigs than in other domestic animals. Two Korean pigs were diagnosed with true hermaphroditism showing ovotestes, epididymes, penes, and uteri. Histomorphologically, the testicular tissues consisted of Sertoli cells that were devoid of spermatogenic germ cells and showed proliferation of interstitial cells. However, the uteri were of normal architecture and had well-developed uterine endometrial glands. The samples were 38, XX female karyotype without the sex-determining region Y (SRY) gene. The findings of this study could contribute to the understanding of true hermaphroditism in the Korean pig industry. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The aim of this study was to examine the effect of acteoside (the cyclin-dependent kinase inhibitor) on the SCNT efficiency with adult fibroblasts in dog. Canine adult fibroblasts were obtained from muscle and cell cycle of fibroblasts was synchronized by culturing to confluency, serum starvation and treating with 30 μM acteoside for 48 h. Cell cycle stages, cell cytotoxicity (apoptosis) and, prduction of reactive oxygen species (ROS) were analyzed using flow cytometry. The canine cells, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were injected into enucleated in vivo matured oocytes, the couplets were electrical fused and activated by calcium ionomycin. SCNT embryos using acteoside-treated fibroblasts were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. In cell cycle synchronization (G0/G1), there was no significant difference between serum starvations (83.9%) and acteoside treated groups (81.3%) that were higher than confluent group (78.5%). In production of apoptosis, confluent and acteoside treated groups (4.3 and 4.5%, respectively) were generated less than serum starvation group (21.8%). In case of ROS, serum starvation group was induced a significantly higher than other groups. After synchronization of the donor cell cycle, either confluent or acteoside treated, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into naturally estrus-synchronized surrogates. Fusion and cleavage rate of acteoside treated group were 64.1 and 41.5%, which were higher than those of confluent group (53.9 and 20.6%, respectively). The reconstructed embryo development rates to 4-cell and 8-cell in acteoside treated group were 29.5 and 14.8%, respectively, while confluent group showed 11.1 and 3.2%, respectively. Total 54 SCNT embryos using acteoside-treated fibroblasts were transferred into oviducts of 2 recipient dogs and one recipient finally delivered one puppy, whereas din`t detected pregnancy on transfer of cloned embryos reconstructed with confluent cells in 6 surrogate dogs. In conclusion, the results of the current study demonstrated that canine fibroblasts could be successfully arrested at the G0/G1 stage with reduced the formation of ROS and apoptosis after acteoside treatment. This results may contribute to improve the effi-ciency of canine SCNT. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The objective of the current study was to describe in vitro embryo production in Hanwoo, analyzing oocytes yield and embryo production. The effects of oocytes production and the number of OPU procedures per animal on embryo production were also evaluated. OPU was done every 3～4 days during experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, we compared the recovery rate of oocytes based on OPU session (Experiment 1). The average of collected oocytes was calculated from every 10 session. The average number of total oocytes recovered per animalonsessionwas 5.16 (mean). Second, we compared the recovery rate base on collection period of OPU (Experiment 2). The following results show the difference of the number of recovered oocytes in every month during the procedure between the months of session. Every animal shows the constant number of recovered oocytes for the first 5 months. However, the recovery rate of oocytes was decreased from month 6 to 8. Third, we compared the developmental rate to blastocyst in two groups (Experiment 3). Oocytes by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24～48 h after in vitro fertilization (IVF) was 75.8% and blastocyst development rate was 18.8% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (61.1%), blastocyst development rate was higher compared with needle puncture group (28.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio- technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being.