In-vitro food mastication has been simulated by incorporating size reduction process, followed by incubation with human saliva. However, they do not suffice to simulate the actual oral breakdown process where disruption by mastication (chewing) and wetting/lubrication by salivation take place simultaneously. In this study, in-vitro oral digestion of rice gels with different amylose contents was simulated by instrumentally performing mastication and salivation. The hardness of artificial saliva-treated rice gels was distinctly lowered with increasing chewing cycles and the rice gels with high amylose contents showed the highest degradation rate. When the rice gels were subjected to non-destructive tomographic analysis, the high amylose rice gels were clearly shown to be favorably fragmented into smaller pieces by chewing, followed by the low and intermediate amylose samples. These results could be explained by their cohesive texture rather than hardness. Therefore, this study may help understanding the changes in the physical properties of cereal-based foods during oral digestion.

This study was conducted to in vitro antioxidant and anti-inflammatory activities of EtOH extracts of Cordyceps militaris (CM). Antioxidant potential, total phenolic and flavonoid contents of CM EtOH extracts were determined by Folin-Ciocalteu method and the aluminum chloride colorimetric method, respectively. Antioxidant activity of CM extracts was measured by following some well-established methods for free radical scavenging such as 2,2-diphenyl-picrylhydrazyl hydrate and 1,2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid). Moreover, Anti-inflammatory activity of CM extracts was determined by measuring the inhibition of nitric oxide (NO) production in lipopolysaccharide/interferon-γ-activated RAW264.7 macrophage-like cells. In addition, cytotoxicity of CM extracts against macrophages was determined by MTT assay. Our results showed that total phenolic content was 19.7 mg gallic acid/g extract. Total flavonoid content was 5.0 mg Naringin/g. Its antioxidant activity was assessed by IC50 value and the values are 338.8 μg/ml (DPPH radical scavenging), 35.4 μg/ml (ABTS radical scavenging). In addition, CM extracts attenuated NO production through the reduction of cellular inducible NO synthase protein expressions. Using MTT assay on indicate that CM extracts showed no toxicity. In conclusion, these results provide important evidence that CM extracts can potentially be used to antioxidant and anti-inflammatory agent.

식물 및 동물성 단백질 유래 펩타이드 형태의 단백질 가수 분해물들은 항산화, 고혈압 완화, 면역조절, 진통완화 및 항균작용 등 생리활성이 있는 것으로 알려져 왔다. 본 연구는 연산오계란 단백질 가수 분해물을 Ultrafiltration를 이용하여 HDS(분획되지 않은 가수 분해물), 1 kDa, 5 kDa, 10 kDa, 50 kDa로 분획된 기능성 펩타이드의 DPPH radical scavenging activity, superoxide anion radical scavenging activity, hydroxyl radical scavenging activity 및 Fe2+ chelation ability을 평가하였다. 그 결 과 DPPH radical scavenging activity 최대값은 1 kDa(70.83 %), hydroxyl radical scavenging activity 최대값은 5 kDa (47.01 %), superoxide anion radical scavenging activity 최대값은 5 kDa(40.57 %), Fe2+ chelation ability 최대값은 5 kDa(29.87 %)로 나타났다. Ultrafiltration를 이용하여 fractionation된 단백질 가수 분해물의 항산화 저해 능력 IC50 평가하였다. 그 결과 HDS의 최대값은 superoxide anion radical scavenging activity(IC50, 5.42 mg/ml)이고, 1 kDa의 최대값은 Fe2+ chelation ability(IC50, 1.67 mg/ml)이고, 5 kDa의 최대값은 Fe2+ chelation ability(IC50, 2.09 mg/ml)이고, 10 kDa의 최대값은 Fe2+ chelation ability(IC50, 2.61 mg/ml)이고, 50 kDa의 최대값은 Fe2+ chelation ability(IC50, 4.53 mg/ml)이 다. 그러므로 본 연구 결과를 바탕으로 5 kDa를 이용하여 오계란 단백질에서 분획한 펩타이드는 항산 화 기능성 식품소재로서 활용할 가치가 높을 것으로 기대한다.

In Korea most bulbs of calla lily (Zantedeschia spp.) are imported from foreign countries including USA and Netherlands due to frequent viral infection problems in field propagation. To solve this problem, in vitro propagation system of calla lilies has been attempted, which ultimately would have an agronomical impact on Korean calla lily farmers. In this study, we carried out experiments to find an efficient method for multiple shoot formation and subsequent root induction in calla lily ‘Black Star’. Lateral buds were used for the induction of multiple shoots on basal MS media containing various concentrations of auxin and cytokinin. The highest callus and multiple shoot formation was observed in MS medium containing 1.0 mg/L of each NAA and BA, in which 25 (83.3%) explants exhibited callus formation and all 30 explants produced multiple shoots with an average of 14.7 shoots per explant in average length of 4.1 cm. Subsequently, obtained shoots were tested for root induction in MS media containing auxins, IAA, NAA, or 2,4-D, because calla lily shoots derived from tissue culture did not produce sufficient roots that are crucial for the survival under natural environment after transplanting. An average of 4.9 or 3.1 roots was generated after 20-day culture of shoots on MS media containing 1.0 mg/L IAA or 1.0 mg/L 2,4-D, respectively. However, no root formation was observed in MS media supplemented with NAA, indicating that NAA is not adequate for root induction of shoots derived from tissue culture of ‘Black Star’.

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.

In order to compare greenhouse gases emission from different animal manures and to explore how different animal manures effect on soil mineralization, three kinds of materials, cattle, goat and chicken manure were amended to soil for 14 days incubation as CtS (cattle manure-amended soil), GS (goat manure-amended soil) and ChS (chicken manure-amended soil). Cumulative NH3 emissions in all treatments were rapidly increased until day 7 and then it was slightly increased in three manure-amended soils but maintained in control until day 14. GS had the highest NH3 emission at 0.14 mg kg-1 during the entire experimental period. Emissions of CO2 were highly increased by 7.8-, 9.0- and 12.4-fold in CtS, GS and ChS, respectively, compared to control at day 14. A significant increase of N2O emission in all treatments occurred within 5 days and then it was slightly increased until day 14. N2O emission was 2-fold higher in all manure-amended soils than that of control. Compared to day 1, inorganic N (NH4 + plus NO3 --N) content was highly increased in all four treatments at day 14. The increase rate was the highest in CtS treatment. Net N mineralization was increased by 4.0-, 2.4- and 2.9-fold in CtS, GS and ChS, respectively, compared to control. These results indicate that increase of NH3, CO2 and N2O gas emissions was positively related to high N mineralization.

This study was conducted to investigate the effects of alpha-lipoic acid (aLA) as an antioxidant that decrease the reactive oxygen species (ROS) in bovine embryonic development. Slaughterhouse derived bovine immature oocytes were collected and 4 different concentrations (0, 5, 10 and 20 mM) of aLA was supplemented in bovine in Vitro maturation (IVM) medium. After 20 hrs of IVM, maturation rates, levels of ROS and glutathione (GSH), and further embryonic development after parthenogenetic activation (PA) and in Vitro fertilization (IVF) was investigated according to aLA concentrations. Maturation rate was significantly higher in 10 mM group than other groups (80.5% vs. 62.9, 73.9, 64.2%; P<0.05). In the levels of ROS and GSH in matured oocytes as an indicator of oocyte quality, significantly better results were shown in 5 and 10 mM groups compared with other 2 groups. After IVM, significantly higher rates of blastocyst formation were shown in 10 mM groups in both of PA (27.9% vs. 18.8, 22.3, 14.2%; P<0.05) and IVF (32.6% vs. 23.9, 27.3, 16.2%; P<0.05) embryos. In addition, significantly more cell total cell number and higher inner cell mass ratio in 10 mM PA and IVP blastocysts showed developmental competence in 10 uM groups. Therefore, based on the entire data from this study, using 10 μM of aLA confirmed to be the optimal concentration for bovine oocyte maturation and embryonic development.

The objective of this experiment was to explore the effects of Roscovitine (Rosco) prior to in vitro maturation (IVM) of immature pig oocyte. Brilliant cresyl blue test has been used to select the good quality of oocyte. Specifically, the effects of Rosco exposure on nuclear and cytoplasmic maturation, diameter, intracellular glutathione (GSH) and reactive oxygen species (ROS), and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT), and gene expression levels in SCNT embryos have been measured. Cumulus oocyte complexes (COCs) have been exposed in 75 μM of Rosco for 22 and 44 h. The COCs that were matured in the IVM for 44 h without Rosco used as control group. Diameter of matured porcine oocytes 44 h culture with Rosco was significantly lower than 22 h culture with Rosco and control groups. GSH was higher in control group than 22 h and 44 h with Rosco but reduction of ROS in 22 h than 44 h with Rosco. In PA, exposure with Rosco 44 h oocytes group has been significantly lower than 22 h and control group in rates of maturation, cleavage and blastocyst formation. Similarly, in SCNT embryos rates of maturation, cleavage and formation of blastocyst have been also significantly lower in 44 h Rosco treated group than other two groups. SCNT embryos treated with Rosco 22 h showed greater expression levels of POU5F1, DPPA2 and NDP52Il mRNA compared with other two groups. Our results demonstrate that Rosco treatment with 22 h prior to IVM improves the development competence of porcine oocyte.

본 연구는 팽이버섯 수확 후 배지를 첨가하여 호밀 사일리지 제조 시 팽이버섯 수확 후 배지를 에너 지원으로서 사용하기 위하여 in vitro 반추위 발효실험이 수행되었다. 공시 사일리지는 출수기의 호밀 에 팽이버섯 수확 후 배지 첨가비율(0%, 20%, 40%, 60%)에 따라 제조하여 6주일간 발효시켰다. In vitro 배양액 제조를 위한 반추위액은 농후사료와 볏짚을 40:60의 비율로 급여한 반추위 cannula가 시 술된 Holstein 수소 2두로 부터 채취하였다. In vitro 실험은 발효시간대를 3, 6, 9, 12, 24 및 48시간 으로 설정하고, 각 처리구별로 3반복으로 발효특성과 건물소화율을 측정하였다. In vitro 배양액의 pH 는 발효시간이 길어짐에 따라 낮아지는 경향이었으며, 48시간 경과 시에는 버섯수확 후 배지 60% 첨가 구가 타 처리구에 비해 유의적(p<0.05)으로 낮았다. 미생물 성장율은 배양시간이 경과함에 따라 증가 하는 경향이었으며, 발효 48시간 경과 시에는 버섯수확 후 배지 20% 첨가구가 타 처리구에 비해 유의 적으로(p<0.05) 높았다. Gas발생량은 48시간 발효 시에 대조구가 타 처리구에 비해 유의적(p<0.05)으 로 높았다. 건물소화율은 버섯수확 후 배지의 첨가비율이 높을수록 높았는데, 발효 24시간 및 48시간에 는 R-60구가 처리구 중에서 가장 높았으며(p<0.05), 대조구에서는 전 발효기간 동안 건물소화율이 현 저히 낮은(p<0.05) 상태에 있었다. In vitro 반추위내 발효실험의 결과와 버섯수확 후 배지의 활용성을 고려할 때, 호밀사일리지 제조 시 팽이버섯 수확 후 배지 첨가비율을 원물기준으로 60%수준이 가장 긍 정적인 것으로 나타났다. 향후 대사시험이나 사양시험을 통하여 가축사료로써의 최적 대체 비율을 규명 해야 할 것으로 사료되는 바다.

To increase the productivity of in vitro development, the antioxidants have been used for culture system of bovine oocytes and embryos. However, comparative studies on these molecules are rare and direct beneficial effects on blastocyst production cannot be discriminated for best results. The study was conducted to determine the influence of N-acetyl-L-cysteine (NAC), N-acetyl-L-cysteine amide (NACA), glutathione (GSH) and cysteamime (CYS) on maturation competence of COCs from GV to MII stage and productivity of blastocyst formation during in vitro fertilization and culture. There was no difference among maturation rates of oocytes to metaphase II with polar body with antioxidants for any of the treatment groups (p>0.05). However, the significant improvement on the rate of blastocysts (32.3±5.0%) was found in 0.1 mM CYS treatment than 0.3 mM NAC, 0.2 mM NACA or 0.5mM GSH (p<0.05). The addition of NAC (18.8±3.7%) or NACA (21.2±3.9%) did not improve development competence to morula and blastocysts than control (24.4±4.1%) and GSH (26.5±5.0%) (p>0.05). Our study showed that medium supplementation with CYS during IVM and IVC improved the rate of bovine embryo development but not with NAC, NACA and GSH addition.

In most mammals, metaphase II (MII) oocytes having high maturation promoting factor (MPF) activity have been considered as good oocytes and then used for assisted reproductive technologies including somatic cell nuclear transfer (SCNT). Caffeine increases MPF activity in mammalian oocytes by inhibiting p34cdc2 phosphorylation. The objective of this study was to investigate the effects of caffeine treatment during in Vitro maturation (IVM) on oocyte maturation and embryonic development after SCNT in pigs. To this end, morphologically good (MGCOCs) and poor oocytes (MPCOCs) based on the thickness of cumulus cell layer were untreated or treated with 2.5 mM caffeine during 22-42, 34-42, or 38-42 h of IVM according to the experimental design. Caffeine treatment for 20 h during 22-42 h of IVM significantly inhibited nuclear maturation compared to no treatment. Blastocyst formation of SCNT embryos was not influenced by the caffeine treatment during 38-42 h of IVM in MGCOCs (41.1-42.1%) but was significantly improved in MPCOCs compared to no treatment (43.4 vs. 30.1%, P<0.05). No significant effects of caffeine treatment was observed in embryo cleavage (78.7-88.0%) and mean cell number in blastocyst (38.7-43.5 cells). The MPF activity of MII oocytes in terms of p34cdc2 kinase activity was not influenced by the caffeine treatment in MGCOCs (160.4 vs. 194.3 pg/ml) but significantly increased in MPCOCs (133.9 vs. 204.8 pg/ml). Our results demonstrate that caffeine treatment during 38-42 h of IVM improves developmental competence of SCNT embryos derived from MPCOCs by influencing cytoplasmic maturation including increased MPF activity in IVM oocytes in pigs.

The elevated temperature and high humidity has been known as main reason for heat stress on animals and cause detrimental effects on productivity of organisms and physiological conditions of normal bioactivities. The aims of this study were to evaluate the relationship between time of heat shock simulation during in vitro maturation and developmental competence of subsequent embryo after in vitro fertilization. Heat shocked cumulus-oocyte complexes (COCs) of Korean native cattle were subjected to normal conditions for 22, 21, 18 and 12 h respectively and transferred to heat stress inducing condition at 40.5 °C in other incubator for 0 (control), 1 and 4 h. After maturation for 22 h, the oocytes were fertilized and cultured in mSOF media for 8 d and examined the developmental capacity of embryos. There were no differences in maturation and cleavage rates between 0, 1 and 4 h heat socked oocytes, but blastocysts formation were lower in the 4 h heat stressed oocytes. The apoptotic cells of developed blastocysts were also increased in at day 8 with 4 h heat shocked oocytes. These results indicate that heat shock on oocytes during maturation could cause negative effects on the developmental competence of embryos.

반하(Pinellia ternata(Thunb.) Breit)는 천남성과에 속하는 다년생 초본식물로서 조직배양을 이용하 여 대량번식 방법 연구가 활발히 진행되어 왔다. 하지만 대량생산된 괴경들의 토양 순화 및 적응을 위 한 환경 조건의 확립의 연구는 미비하다. 따라서 본 연구에서는 기내에서 증식된 반하의 괴경을 이용하 여 우수한 품질의 약재와 건전묘의 대량생산을 위해 생육에 적합한 토양조건을 탐색하였다. 본 연구에 서는 액체배지에서 현탁배양 한 괴경(Type 1)과 고체배지에 치상하여 배양된 괴경(Type 2)두 종류를 비 교하였다. 토양은 3개의 조성으로 조합하여 생육을 비교하였으며, 코코피트, 피트모스, 버미큘라이트, 펄라이트 및 제오라이트를 배합하여 사용하였다. 기내 배양 조건이 다른 처리구들의 토양 조성별 순화 율 측정하였으며, 생육의 차이를 확인하기 위해 8주동안 생육 후 초장, 잎 수, 마른잎 수, 괴경 수, 괴 경 크기, 생체중 및 건물중을 측정하였다. 또한 주아의 생성율을 확인하기 위하여 4주와 8주에 측정을 진행하였다. 그 결과 Type 2가 펄라이트를 20%증량한 상토 B에서 가장 우수한 생육을 보였다. 괴경의 비대에 영향을 주고 묘로서 이용을 위하여 필요한 잎의 수는 상토 B에서 가장 많은 1.7개로 나타났고 가장 잎의 출현이 없었던 Type 1의 상토 C가 0.9개로 나타나 1.8배의 차이를 나타냈다. 또한 같은 처 리구에서 건물중이 통계적으로 유의한 차이를 보이며 우수한 것으로 나타났다. 반하의 주아 생성율의 차이는 상토 B에서 1.1개로 우수하였으며, 가장 저조한 처리구의 수치보다 1.2배 높은 생성율을 확인할 수 있었다. 이러한 결과로 기내에서 배양된 반하의 토양 순화 및 적응기에 괴경의 비대를 유도하여 우 수한 한약재 생산이 가능 할 것으로 생각되며 차후 토양에서 재배 시 반하의 대량번식에도 도움이 될 것이라 사료된다.

The purpose of this study was to investigate and analyze the total phenolic content (TPC), antioxidant activities (FRAP, ABTS, and DPPH), and antibacterial properties of lotus (Nelumbo nucifera) extracts. Lotus leaves, stems, and seed pods were extracted with deionized water at 95℃, and with 70.5% ethanol at 85℃. The TPC ranged from 8.12 to 215.12 GAE mg/g. The ethanol extract of the seed pod had the highest TPC, and the TPC of the corresponding deionized water extract was 161.45 mg/g. FRAP values ranged from 104.03 to 3,546.39 TEAC μmol/g, ABTS radical cation scavenging activities ranged from 105.11 to 3,956.94 TEAC μmol/g, and DPPH radical scavenging activities ranged from 37.29 to 2,549.46 TEAC μmol/g. EC50 values ranged from 0.26 to 9.63 mg/mL, and 0.31 to 21.21 mg/mL for ABTS and DPPH, respectively. The ethanol and deionized water extracts of the seed pod showed higher TPC and stronger antioxidant properties (FRAP, ABTS, and DPPH) than those of characteristic of the leaf extracts. The ethanol and deionized water extracts of the seed pod showed antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa with inhibition zones of 9.0 to 14.0 mm, and the ethanol extract of the leaf showed antimicrobial activity against B. subtilis and S. aureus with inhibition zones of 9.0 and 10.0 mm, respectively. Thus, the lotus seed pod could be used to produce novel teas, and could be a potential source of therapeutic ingredients for food and medicine.

This study was aimed to evaluate the dose-response the effects of nano-encapsulated conjugated linoleic acids(CLAs) on in vitro ruminal fermentation profiles. A fistulated Holstein cow was used as a donor of rumen fluid. Nano-encapsulated CLAs(LF, 5% of nano-encapsulated CLA-FFA; HF, 10% of nano-encapsulated CLA-FFA; LT, 5% of nano-encapsulated CLA-TG; HT, 10% of nano-encapsulated CLA-TG) were added to the in vitro ruminal fermentation experiment. In the in vitro ruminal incubation test, the total gas production on incubation with nano-encapsulated CLAs was increased significantly according to the incubation time, compared with the control(p<0.05). The tVFA concentrations on addition of LF and HT were significantly higher than that of the control(p<0.05). Thus, nano-encapsulated CLAs might improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, the population of Butyrivibrio fibrisolvens which is closely related to ruminal biohydrogenation was increased by adding HT, while decreased by adding LF at 12 h incubation. These results showed that nano-encapsulated CLA-FFA could be applied to enhance CLA levels in ruminants by maintaining the stability of CLA without causing adverse effects on ruminal fermentation profiles considering the optimal dosage.

본 시험은 항염에 효과가 있다고 알려진 식물 추출물의 첨가가 반추위 발효와 메탄 생성에 미치는 영 향을 알아보기 위해 in vitro 실험을 수행하였다. 항염과 항산화 효과가 있는 산뽕나무, 뽕나무, 예덕나 무, 오동나무, 방아풀과 은행나무 6종을 선발하였다. 반추위액과 McDougall buffer 혼합액 15mL과 티 모시 0.3g, 각각의 추출물을 기질의 5%로 넣고 39℃에서 3, 9, 12, 24, 48 그리고 72시간 배양하였다. 항염 효과가 있는 식물 추출물의 첨가는 반추위 발효 성상(pH, 건물소화율, Glucose 농도, 암모니아 농도, 단백질 농도, 미생물 성장량, 휘발성지방산)에는 영향을 미치지 않았다. 총 가스 발생량은 각기 다른 양상을 보였으며, 이산화탄소 발생량은 48시간대에서 예덕나무와 방아풀에서 대조구에 비해 유의 적(p<0.05)으로 높았다. 또한 메탄 발생량은 배양 초기에는 대조구보다 첨가구에서 유의적(p<0.05)으로 감소하였으나, 발효가 진행될수록 대조구에 비해 첨가구에서 더 많은 메탄이 발생하였다. Polyphenol과 flavonoid는 은행나무 추출물구에서 가장 높았다. 본 시험의 결과에서 항염에 효과가 있는 식물 추출물 을 in vitro 반추위 배양액에 첨가하였을 때, 반추위 발효에는 영향을 미치지 않았고, 메탄은 초기 발효 에 저감 효과가 있는 것으로 사료된다.