본 실험에서는 외인성 효소 첨가제 및 혼합 세균 배양을 통한 고상발효(Solid-state fermentation, SSF)가 채종박(Rapeseed meal, RSM)의 체외건물소화율(In vitro dry matter digestibility, IVDMD) 및 단쇄지방산(Short chain fatty acid) 생성에 미치는 영향을 조사하기 위해 수행되었다. 외인성 효소 칵테일(첨가 및 미첨가) 및 RSM에 대한 SSF(발효 및 비발효)를 나타내는 2 x 2 요인 설계가 적용되었다. 3-step 돼지 소화율 모델을 적용하여 채종박의 건물 소화율을 분석하였으며, 72시간 대장발효 후 상층액을 수집하여 단쇄지방산 생성량을 분석한 후 칼로리 단위로 변환하여 가소화에너지 소화율을 분석하였다. 소장 (IVDMDh) 및 전장 (IVDMDt) 건물소화율에서는 고상발효된 채종박이 더 높게 나타났다 (각각 p < 0.01). 마찬가지로, 외인성 효소 첨가제 처리구에서 채종박의 소장 소화율(IVDMDh)이 증가하는 경향을 나타냈다(p = 0.06). Acetic acid 및 butyric acid의 생산은 대조구에 비해 고상발효 처리 시 유의하게 더 생산되었으며 (각각 p < 0.01), 이는 총 단쇄지방산의 생산 증가 경향을 나타냈다(p = 0.09). 에너지 소화율에서는 채종박의 고상발효 및 외인성 효소제 첨가가 유의적으로 높게 나타났다 ( p < 0.01). 그러므로 채종박의 고상발효 처리는 단백질 이용성을 비롯한 영양적 가치를 향상시키는데 효과적이라고 사료된다.

This study is concerned with the optimization of the manufacturing process of a hot water extract containing antioxidant activity from Lycium barbarum, traditionally known to have various physiological activities. For the establishment of the optimization process, the central composite design of response surface methodology(RSM) was used. Thirteen extraction processes were performed by encoding the independent variables, extraction temperature (65.9oC–94.1oC) and extraction time (2.59 hr–5.41 hr). As a result of the experiment, the optimal manufacturing conditions for the extract were 340.0 mg/100 g of GAE at an extraction temperature of 94.1oC and an extraction time of 5 hr. The maximum yield of flavonoids was 22.44 mg/100 g of HES at an extraction temperature of 94.1oC and an extraction time of 4 hr. The conditions for producing the extract with the maximum antioxidant capacity (DPPH 92.12%) were 90oC and 4.5 hr extraction time. Therefore, the optimal manufacturing process conditions for extracts containing total phenol content, flavonoid content, and DPPH radical scavenging activity, which are dependent variables, were extraction temperature of 90-95oC and extraction time of 4 hr, which were not significantly different from the actual values. Therefore, Lycium barbarum extract rich in total phenol and flavonoid content related to antioxidant function is expected to be used as a functional food and cosmetic material.

Despite numerous advances in in-vitro embryo production (IVP), many documented factors have been shown to influence the development of mammalian preimplantation embryos and the success of IVP. In this sense, elevated levels of reactive oxygen species (ROS) correlate with poor outcomes in assisted reproductive technologies (ART) due to oxidative stress (OS), which results from an imbalance between ROS production and neutralization. Indeed, excessive production of ROS compromises the structural and functional integrity of gametes and embryos both in vivo and in vitro. In particular, OS damages proteins, lipids, and DNA and accelerates cell apoptosis. Several in-vivo and in-vitro studies report an improvement in qualityrelevant parameters after the use of various antioxidants. In this review, we focus on OS and the source of free radicals and their effects on oocytes, sperm, and the embryo during IVP. In addition, antioxidants and their important role in IVP, supplementation during oocyte in vitro maturation (IVM), in vitro culture (IVC), and semen extenders were discussed. Nevertheless, various methods for determining the level of ROS in germ cells have been briefly described. Still, it is crucial to develop standardized antioxidant supplement systems to improve overall IVP success. Further studies should explore the safety, efficacy, mechanism of action, and combination of different antioxidants to improve IVP outcomes.

본 연구는 조사료 원료인 마늘 껍질의 사료적 가치와 메탄 발생량을 조사하기 위해 in vitro 발효 실험을 수행하였다. Garlic husk를 제외하고 국내에서 널리 사용되는 4가지 조사료 원료(Oat hay, Annual ryegrass, Timothy, Tall fescue)를 실험의 처리구로 구성하였다. In vitro 48 시간 발효 후 건물 소화율, 섬유소 소화율, 암모니아태 질소, 휘발성 지방산, pH, 메탄 발생량을 평가하였다. 가스 발생량은 3, 6, 9, 12, 24, 36 및 48 시간에 각각 측정되었다. 최종 가스 발생량에서 Oat hay가 유의적으로 가장 높았다(p<0.01). Garlic husk는 처리구 중 가장 높은 메탄 발생량을 나타내었다(p<0.01). Garlic husk의 건물 소화율은 Oat hay보다 유의적으로 낮았지만 Annual ryegrass, Timothy, Tall fescue보다 높았다(p<0.01). 섬유소 소화율은 Annual ryegrass에서 가장 낮았다(p<0.01). 총 휘발성 지방산 생성량은 Oat hay에서 가장 높았고(p<0.01), Garlic husk, Timothy, Tall fescue는 유사하였다. 본 연구의 in vitro 발효 성상과 메탄 발생량을 고려할 때, Garlic husk는 국내 주요 조사료를 대체할 영양적 가치를 포함하고 있으나 메탄 저감 효과는 미미할 것으로 추정된다.

The objective of this study was to evaluate effects of feeding methods on in vitro ruminal fermentation, total gas and methane production in Hanwoo steers. Six Hanwoo steers fitted with rumen cannula (430 ± 21 kg of body weight) were randomly assigned to one of three feeding systems: 1) feeding forage 1 hour after concentrate, 2) feeding concentrate 1 hour after forage, 3) feeding mixed ration. Rumen fluid sampled from each animals was incubated 24 hours with maize or timothy substrates in in vitro. Ruminal pH was increased in feeding method 2 or maize substrate than that of other methods or timothy substrate (P < 0.001). The production of total volatile fatty acid, acetate, propionate, butyrate, and valerate were increased when steers fed diets using feeding method 1 or rumen fluid was incubated with maize substrate (P < 0.001). Increased production of total gas and methane was observed in feeding method 1 and maize substrate compared to those of other methods or timothy substrate (P < 0.001). Due to the inconsistent results between ruminal fermentation and gas production in this study, further research is required to estimate effects of feeding method on enteric fermentation and gas production in in vivo.

The Dromedary camel (Camelus dromedaries) is an important species because of its ability to produce good quality meat, milk, and fibers under harsh environmental conditions. Camels are also crucial for transportation, racing, and as draft animals in agriculture. Therefore, dromedary camels play a critical role in the economy for millions of people living in the arid part of the world. The inherent capability of camels to produce meat and milk is highly correlated with their reproductive performance. Compared with other domestic species, the reproductive efficiency in camelids is low. Although recent reproductive technologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) have been successfully applied to camelids and the birth of live offspring following these technologies has been reported; in vitro embryo production (IVP) has lagged in this species. The development of the IVP system for dromedary camels may be a useful tool for the genetic improvement of this species. IVP in farm animals includes three main steps; in vitro maturation (IVM) of an oocyte, IVF of a matured oocyte, and in vitro culture (IVC) of fertilized oocyte up to the blastocyst stage. This review aims to summarize various factors that influence oocyte quality, IVM, and in vitro embryo development in dromedary camel.

Lilium dauricum is a rare and endangered species belonging to the family Liliaceae. The species contains several bioactive compounds used as functional foods and medicinal agents in Northeast Asia. This study aimed (1) to establish an in vitro bulblet culture using an air-lift bioreactor and callus culture for the conservation of L. dauricum and obtaining its bioactive compounds; (2) investigate the plant phenolic compounds from both cultures system. The highest bulblet production with 12.5-fold increase in growth rate was obtained using MS medium supplemented with 0.5 g L-1 BA and 3% sucrose. Addition of 7% sucrose facilitated bulblet enlargement, with approximate 2.5- and 7-fold increases in diameter and fresh weight, respectively. The highest rate of callus (100%) was obtained using a combination of 1.0 mg L-1 picloram and 0.5 mg L-1 Kinetin. The callus proliferation occurred on MS medium supplemented with 1.0 mg L-1 picloram, 0.25 mg L-1 kinetin, and 0.25 g L-1 casein hydrolysate. There was a significant difference in the total phenolic compound content of callus, which was 1.5-fold higher than that in the bulblets. These findings indicate a suitable system for optimizing both bulblet and callus culture of L. dauricum, therefore, providing useful bio-materials for industrial purposes and contributing to the conservation of this species.

Although somatic cell nuclear transfer (SCNT)-derived embryonic stem cells (ESCs) in pigs have great potential, their use is limited because the establishment efficiency of ESCs is extremely low. Accordingly, we tried to develop in-vitro culture system stimulating production of SCNT blastocysts with high performance in the colony formation and formation of colonies derived from SCNT blastocysts for enhancing production efficiency of porcine ESCs. For these, SCNT blastocysts produced in various types of embryo culture medium were cultured in different ESC culture medium and optimal culture medium was determined by comparing colony formation efficiency. As the results, ICM of porcine SCNT blastocysts produced through sequential culture of porcine SCNT embryos in the modified porcine zygote medium (PZM)-5 and the PZM-5F showed the best formation efficiency of colonies in α-MEM-based medium. In conclusion, appropriate combination of the embryo culture medium and ESC culture medium will greatly contribute to successful establishment of ESCs derived from SCNT embryos.

Up-to-date artificial insemination (AI) using frozen sperm consider as the most widely using technology for improvement of Korean Native Cow (Hanwoo) embryo production. However, it is time consuming, required at least 15~20 years to make more than 6 generations, and their offspring number is limited. To overcome such limitations, superovulation and in vitro fertilization have been developed. For superovulation, the number of produced embryos are not enough for commercialization and donor cows need rest period. This led to use of slaughterhouse ovary for in vitro fertilization, but it is impossible to repeat the collection from the same individual and it only can improve the genetic merits of offspring for one generation. Production of embryos using Ovum Pick-Up (OPU) technique, where oocytes can be repeatedly collected from living elite donor, might overcome these limitations. In this study, we investigated the possibility of using OPU technique from donors at different age and different session periods for mass-embryo-production. Oocytes were collected from 26 donor cows twice per week, 3 - 4 months per year, between 2013 and 2016. Results showed that, the average number of embryo produced in first year used donor was significantly higher than that in second year used donor (3.89 ± 2.85 vs 3.29 ± 2.70), however, there was no significant difference between third year used donor (3.51 ± 3.32) and other groups. Taken together, our data showed that repeated using of donor up to three years is possible for in vitro embryo mass-production. Moreover, OPU can be used as suitable embryo producing technique for livestock breed improvement.

This study was conducted with two ruminally cannulated Holstein steers to examine the effect of micronized and steam flaked corn on ruminal fermentation characteristics. The in situ dry matter degradability after 48 h incubation was the highest (P<0.05) at micronized corn (2.5 mm thickness) compared with steam flaked corn treatments. The steam flacked corn (3.3 mm thickness) was degraded lower (P<0.05) than the 2.9 and 3.1 mm thickness of steam flacked corn. Effective dry matter degradability and the rate of constant were the highest (P<0.05) at micronized corn (2.5 mm thickness) compared with steam flaked corns as well. The in vitro dry matter degradability after 48 h incubation was tended to higher (P=0.088) at micronized corn (2.5 mm thickness) than steam flaked corns, whereas there is no significantly difference between steam flaked corn treatments. Total volatile fatty acid concentration was higher at steam flaked corn (2.9 mm thickness) than micronized corn (2.5 mm thickness) and steam flaked corn (3.1 and 3.3 mm thickness). The acetate : propionate ratio was the highest (P=0.008) at steam flaked corn (2.9 mm thickness) and the lowest (P=0.008) at micronized corn (2.5 mm thickness). Total gas and methane production after 48h ruminal incubation was the highest (P=0.001) at micronized corn (2.5 mm thickness) compared with steam flaked corns. According to these results, the thickness of steam flaked corn as resulted corn processing is believed to do not affect methane production. However, further study is needed to better understand the present results to verify the correlation between corn processing method and their thickness on methane production using the same thickness corns by difference processing methods.

This study was conducted to examine the effect of corn (Zea mays L.) - soybean (Glycine max L.) silage prepared by intercropping method on the nutritive value of the silage, in vitro rumen fermentation characteristics, dry matter degradability, as well as milk yield and milk composition of dairy cows. In a couple of experiments intercropped corn-soybean silage (CSBS) was compared with corn silage (CS) and/or Italian ryegrass hay (IRG). Numerically, CSBS had higher crude protein, ether extract, and lactic acid contents compared to CS. In vitro rumen fermentation analysis demonstrated that up to a 24-h incubation period, both CS and CSBS showed higher total gas production, ammonia N concentration, and dry matter degradability compared to IRG (p<0.05). The investigation on animals was conducted in a commercial dairy farm located in Gyeongju, South Korea, employing 42 Holstein cows that were divided into 2 group treatments: CS and CSBS in a completely randomized design. Although no significant difference was observed in milk yield, animals fed on CSBS showed significantly higher milk protein (p<0.05) and milk fat content (p<0.01), compared to animals fed on CS. Taken together, our findings indicate that corn-soybean silage that is cultivated, harvested, and prepared through intercropping can improve the protein content of the silage, and can also enhance in vitro rumen fermentation, dry matter degradability, and performance of dairy cattle.

하수오(Polygonum multiflorum Thunb.)는 여뀌과에 속하는 초본식물로 동아시아와 북아메리카에 자생하고 있다. 하수오는 관상용으로도 이용되며, 한의학에서 뿌리를 약용으로 사용한다. 하지만 월동 이 어렵고 노동력 소모가 많아 최근 재배농가가 감소하고 있다. 본 연구에서는 하수오의 우량종묘를 짧 은 시간에 대량생산하기 위하여 수경재배에 적합한 양액과 조직배양묘의 크기를 조사하였다. 토양에 적 응이 완료된 하수오 종묘는 크기에 따라 Type 1, Type 2 및 Type 3으로 나누어 4종류의 양액에 치상 하였다. 치상 후 양액과 종묘에 따른 생육차이를 확인하기 위하여 생육조사를 실시하였다. 생육조사는 8주 동안 양액에서 생육시킨 후 줄기길이, 잎 수, 마른 잎 수, 줄기 수, 생체중과 건물중에 대해 측정 하였다. 그 결과 양액3에서 Type 1과 Type 2의 뿌리길이는 각각 33.8cm와 29.5cm로 각각 나타났다. 이와 유사한 경향으로 덩굴 길이도 CaCl2가 첨가 된 양액3에서 Type 1과 Type 2가 가장 길게 나타나 우수한 생육을 보였다. 이처럼 뿌리가 증대된 종묘는 밭으로 이식 될 때 활착을 도와 우수한 초기생육 을 나타낼 것이라 사료된다. 반면 Type 3는 잎끝마름과 고사하는 현상을 보여 추후 양액의 적정농도에 대한 추가 실험이 필요할 것이라 사료된다.

지황은 현삼과(Scrophulariaceae)에 속하는 다년생 초본식물 Rehmannia glutinosa(Gaertn.) DC.로 서 한의학에서는 뿌리를 지황(地黃)이라는 한약재로 이용하고 있다. 지황은 선지황(鮮地黃), 생지황(生 地黃), 숙지황(熟地黃) 등으로 널리 사용되고 있지만 종자파종에 의한 재배보다 종근을 이용한 영양번식 으로 재배하고 있다. 종근을 이용한 재배법은 뿌리 썩음병으로 인해 종근의 관리와 생산성이 높은 고품 질의 지황 생산이 어려운 실정이다. 본 연구에서는 형태적 특징과 유전자 분석을 통해 기원이 검증된 지황 기원식물을 이용하여 정확한 기원의 무균 배양묘와 종근 생산을 위한 최적 조건을 확립하고자 하 였다. 기내배양묘 생산에 필요한 최적의 생장율과 다개체 형성율, 그리고 뿌리 형성율 및 비대조건을 탐색한 결과, WPM 배지와 1.0mg/L의 IAA와 0.5mg/L의 IBA 옥신계 생장조절제가 첨가된 배지(WPM + IAA 1.0mg/L + IBA 0.5mg/L)에서 최적 생장율과 뿌리다개체 형성율을 나타내었으며, 0.1mg/L의 NAA가 첨가된 WPM 배지(WPM + NAA 0.1mg/L)에서 가장 많은 수의 뿌리가 형성되었고 비대율도 높 게 나타났다. 이러한 최적조건에서 생산한 기내식물체는 토양 순화 후 정식하였을 경우에도 높은 뿌리 형성율과 지속적인 비대 효과를 보였다. 이상의 결과는 지황의 무균 배양묘 생산과 배양묘기반 종근 생 산을 통해 지황 대량 생산에 필요한 배양묘와 종근 생산에 유용하게 활용될 수 있을 것으로 판단되며, 고품질 지황 생산량에기여할 것으로 사료된다.

In vitro production of mammalian embryos has been achieved with the oocytes derived from middle-size follicles (MF, mainly 3-6 mm in diameter) in many species including domestic animals. In the ovaries, however, there are more small-size follicles with less than 3 mm in diameter (SF). If we can develop an efficient system to produce embryos in vitro from the oocytes from SF. In this presentation, I would like to review about embryo production in vitro from the oocytes derived from SF. As well as the diameter of oocytes, the number of cumulus cells surrounding the oocyte derived from SF is significantly smaller those of oocytes from MF. The comparative analysis in electrophoresis about secretions of cumulus-oocyte complexes derived from SF and MF demonstrated a significant difference in the proteins with a molecular weight. Proteins secreted from cumulus cells, vascular endothelial growth factor (VEGF), are 34- to 42-kDa proteins, including seven family members. The molecular weight of VEGF was similar with the secretion we observed. Supplementation of medium for in vitro maturation with VEGF significantly improved the oocytes competence not only to complete the meiosis in vitro but also to develop to the blastocyst stage following parthenogenetical activation. Removing cumulus cells 20 h after the start of culture for in vitro maturation also significantly improved the competence of oocytes derived from SF to achieve the meiosis. A combination of these new techniques may improve more the meiotic and developmental competences.