본 연구는 국화과 추출물 첨가가 in vitro 반추위 발효성상 및 메탄 발생에 미치는 영향을 구명하고자 수행하였다. 반추위액은 cannula 장착된 한우 암소 1두에서 채취하였으며, 공시축의 사양관리는 timothy와 농후사료를 6:4 비율로 하였다. 배양액은 Mcdougall buffer 10mL, 위액 5mL 섞어 50mL serum bottle에 혐기상태로 분주하였다. 티모시 0.3g를 각각 넣고 국화과 추출물인 국화, 민들레, 씀바귀, 제비쑥, 해바라기를 기질의 5%를 첨가한 뒤 발효시간대별(3, 6, 9, 12, 24, 48 및 72시간)로 7처리 3반복 수행하였다. 배양액 pH값은 6.29~7.44으로 반추위 적정 pH범위에 속하였다. 건물 소화율은 발효시간대가 늘어날수록 처리구에서 대조구에 비해 유의적(p<0.05)으로 증가하였다. 총가스 발생량은 발효 48시간대에서 민들레, 씀바귀, 제비쑥, 해바라기에서 대조구에 비해 유의적(p<0.05)으로 높게 측정 되었다. 이산화탄소 발생량은 발효 48시간대에서 민들레, 씀바귀, 제비쑥, 해바라기에서 대조구에 비해 유의적(p<0.05)으로 높게 측정 되었으며, 메탄 발생량도 발효 48시간대에서 민들레, 씀바귀, 제비쑥, 해바라기에서 대조구에 비해 유의적(p<0.05)으로 높게 측정 되었다. 미생물 성장량은 발효 48시간대에서 제비쑥과 해바라기 처리구에서 대조구에 비해 유의적(p<0.05)으로 높게 측정되었다. 발효 48시간대 에서 Acetic acid 생성량이 민들레, 씀바귀, 제비쑥, 해바라기 처리구에서 대조구에 비해 유의적(p<0.05)으로 높았다. 발효 12시간대에서 Propionic acid 생성량은 씀바귀, 제비쑥, 해바라기 처리구에서 대조구에 비해 유의적(p<0.05)으로 높았다. 발효 24시간대에서 A/P ratio는 민들레, 제비쑥, 해바라기 처리구에서 대조구에 비해 유의적(p<0.05)으로 감소하였다. 본 연구의 결과에 의해 국화과 추출물 첨가가 반추위 발효에 부정적인 영향을 미치지 않는 것으로 생각되나 메탄 발생량을 감소시키는데 영향이 없는 것으로 생각된다.

본 연구는 탄닌 함유 식물 추출물을 이용하여 in vitro 반추위 발효성상, 미생물 성장 및 메탄생성에 미치는 영향을 알아보고자 수행하였다. 반추위액은 농후사료와 조사료(timothy)를 40:60의 비율로 급 여한 반추위 cannula가 시술된 한우 암소에서 채취하였다. 본 실험에 사용한 식물 추출물은 상수리나 무(Quercus acutissima), 밤나무(Castanea crenata), 참가시나무(Quercus salicina), 졸참나무 (Quercus serrata) 그리고 떡갈나무(Quercus dentata)를 사용하였으며, 식물 추출물은 한국식물추출물 은행에서 분양받아 사용하였다. 반추위액과 McDougall buffer를 1:2의 비율로 혼합한 배양액을 0.3g timothy와 식물 추출물(기질의 5%)이 담긴 50mL serum bottle에 혐기상태로 15mL를 분주하였다. Serum bottle은 39℃, 120rpm으로 3, 6, 9, 12, 24, 48 및 72시간 동안 배양하였다. 본 실험 결과 배 양 시간이 지날수록 pH는 점점 감소하였으며, 정상범위에 포함되었다. 건물소화율은 대조구에 비해 첨 가구에서 유의적(p<0.05)으로 낮았다. 암모니아, 총 가스 생산량 및 이산화탄소 발생량은 대조구에 비 해 첨가구에서 유의적(p<0.05)으로 차이가 없었으나, 24시간 메탄 발생량은 대조구에 비해 전 첨가구에 서 유의적(p<0.05)으로 낮았다. 미생물 성장량은 첨가구에 따라 각각 다른 양상을 나타내었고, acetic acid 및 propionic acid는 대조구에 비해 첨가구에서 유의적(p<0.05)으로 낮았다. Real-Time PCR에서 섬유소 박테리아(R. albus, R. flavefaciens) 및 메탄 생성균(Ciliate associated methanogen)은 대조 군에 비해 유의적(p<0.05)으로 감소하였다. 결과적으로 본 실험에 사용한 탄닌 함유 식물 추출물 5종 모두 반추위 발효에 영향을 미치지 않으며, 메탄저감 효과를 나타내었다. 특히 상수리나무에서 반추위 발효성상에 악영향을 미치지 않으며 메탄 발생을 저감하는 식물 추출물로 적합하다고 생각된다.

연속류도로 합류영향구간에서는 본선과 다른 교통특성을 가진 연결로교통류의 합류로 차로변경 및 가감속과 같은 차량간의 불규칙한 상호작용이 발생하여 교통류의 난류현상을 야기한다. 따라서, 난류현상은 운행상태를 고려하여 연속된 지점간의 불안정한 교통특성으로 판단하여야 한다. 본 연구에서는 합류영향구간에 차로-지점별 검지기를 설치하여 시공간적으로 연속된 교통자료를 구축하였으며, 지점간에 유의한 속도변화를 판단하는 기준으로 최소유의차(LSD) 통계값을 산정하여 난류현상을 분석하였다. 분석결과, 합류영향구간 난류현상은 운행상태에 따라 발생권역 및 심각도가 변화되는 것으로 나타났다. 이에 따라 난류현상에 의한 최대 합류영향권역은 교통량이 증가하는 혼잡전 운행상태에서 보여지며, 속도변화특성에 따라 상류 100m~하류 100m의 "감속구간"과 하류 100m~하류 400m의 "감속속도유지 및 가속구간"으로 구분할 수 있었다.

This study was conducted to evaluate roughage to concentrate ratio on the changes of productivity and metabolic profiling in milk. Six lactating Holstein cows were divided into two groups, T1 group was fed low-concentrate diet (Italian ryegrass to concentrate ratio = 8:2) and T2 group was fed high-concentrate diet (Italian ryegrass to concentrate ratio = 2:8). Milk samples were collected and its components and metabolites were analyzed by 1H-NMR (Nuclear magnetic resonance). The result of milk components such as milk fat, milk protein, solids-not-fat, lactose and somatic cell count were not significantly different between two groups. In carbohydrate metabolites, trehalose and xylose were significantly higher (P<0.05) in T1 group, however lactose was not significantly different between two groups. In amino acid metabolites, glycine was the highest concentration however, there was no signifi-cant difference observed between two groups. Urea and methionine were significantly higher (P<0.05) in the T2 group. In lipid metabolites, carnitine, choline and O-acetylcarnitine there were no significant difference observed between the two groups. In benzoic acid metabolites, tartrate was significantly higher (P<0.05) in T2 group. In organic acid metabolites, acetate was significantly higher (P<0.05) in T1 group and fumarate was significantly higher (P<0.05) in T2 group. In the other metabolites, 3-methylxanthine was only significantly higher (P<0.05) in T2 group and riboflavin was only significantly higher (P<0.05) in T1 group. As a result, milk components were not significantly different between two groups. However, metabolites concentration in the milk was significantly different depends on roughage to concentrate ratio.

This study was conducted to investigate for the natural methane emission inhibitor as a feed additive no adversely effect on rumen fermentation. Five different Control (Wheat barn (0.05 g), MRA(Methane Reduction Additive)-1 (Allium fistulosum L. (0.05 g)), MRA-2 (Sodium Lauryl Sulfate (0.025 g) +Wheat barn (0.025 g) mixed), MRA-3 (Sodium Dodecyl Sulfate (0.025 g) + Wheat barn (0.025 g) mixed), and MRA-4 (Allium fistulosum L. (0.02 g) + Tannic acid (0.02 g) + Wheat barn (0.01 g) mixed) contents were used to perform 3, 6, 9, 12, 24 and 48 h incubation for in vitro fermentation. Ruminal pH values were ranged within normal ruminal microbial fermentation. Dry matter digestibility was not significantly different across the treatments during the whole fermentation time. Also, the result of microbial growth had no adversely effect on during the whole fermentation time. At 24 h, methane emission was significantly lower (P<0.05) than all treatments except to MRA-1. Especially, MRA-4 carbon dioxide emission was significantly lower (P<0.05) than control at 9, 24 and 48 h incubation. In addition MRA-4 propionate concentration was significantly higher (P<0.05) than control at 24 h incubation. The result of RT-PCR Ciliate-associated methanogens were significantly lower (P<0.05) at MRA-1, MRA-3 and MRA-4 than control at 24 h incubation. Based on the present results, MRA-4 could be suggestible methane emission inhibitor as a natural feed additive.

This study was conducted to evaluate Raphanus sativus extracts to methane reduction in rumen. Five different levels of R. sativus extracts were used to investigate the most effective dosing level for the decrease of methane production in the rumen. The rumen fluid was collected from a cannulated one Hanwoo cow (BW=450±30 kg) consuming 600 g/kg timothy and 400 g/kg concentrate. On fermentation day, rumen fluid was collected at 2 hr postfeeding R. sativus extracts was dosed to achieve final concentration of 0, 1, 3, 5, 7, and 9% respectively, to fermentation bottles containing the mixture of rumen fluid and McDougall’s buffer and 300 mg of timothy was added as a substrate. The fermentation was conducted for 3, 6, 9, 12, 24, 48 and 72 hr incubation time at 39℃ with shaking. In vitro ruminal pH values were measured normal range for ruminal fermentation. Dry matter disappearance was significantly higher (p<0.05) at 3 hr incubation time 1, 3 and 5% doses than that of control. The highest methane reduction was observed in 12 hr incubation time 5, 7 and 9%. The carbon dioxide emission was also significantly (p<0.05) lower than that of control at 12 hr incubation time 5, 7 and 9%. The total volatile fatty acid was no significant difference between control and all doses level at 12 and 24 hr incubation time. At 24 hr incubation time, the result of real-time PCR were indicated that M. archea was significantly lower (p<0.05) at all doses level comparing to that of control. In conclusion, R. sativus extracts were significantly decreased methane emission. R. sativus extracts were significantly lower (p<0.05) than that of control at 12 hr incubation time 5, 7 and 9% and no adversely effect in rumen pH, dry matter disappearance and total VFA.

This study was conducted to evaluate natural plant extracts for methane gas reduction in ruminants. Rumen fluid was collected from cannulated Hanwoo cow (450±30 kg) consuming 400 g/kg concentrate and 600 g/kg timothy. The 15 ml of mixture comparing McDougall’s buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically into 50 ml serum bottles. Rumen fluid contents were collected and in vitro fermentation prepared control (timothy, 300 mg), ginseng, balloon flower, yucca plant, camellia, tea plant and ogapi extracts were added at the level of 5% against 300 mg of timothy as a substrate (v/w) and incubated for 3, 6, 9, 12, 24, 48, and 72 h. In vitro pH values range 6.55~7.41, this range include rumen titration. The dry matter digestibility was not differ between all treatments and control. Total gas emission was significantly higher (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Carbon dioxide emission was not differ all treatments on 9 h than in control and significantly higher (p<0.05) yucca plant, camellia and tea plant treatments on 12 h than control. Methane emission was not differ all treatments on 6 h than in control. The rumen microbial growth rate was significantly higher (p<0.05) in ginseng, balloon flower on 12 h and significantly higher (p<0.05) in ginseng, yucca plant, tea plant and ogapi treatments on 24 h than in control. Total VFA was significantly higher (p<0.05) in tea plant and ogapi treatments on 12 h than in control and significantly higher (p<0.05) in ginseng, balloon flower treatments on 48 h than in control. Acetic acid was significantly lower (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Propionic acid was significantly higher (p<0.05) in ginseng and balloon flower treatments on 48 h than in control. As a results, sixth natural plant extracts had no significant effect dry matter digestibility and negative on rumen fermentation, but not effect methane reduction.

To develop environment-friendly agricultural products with anti-microbial activity against Sclerotinia sclerotiorum as a pathogen of sclerotium disease, Aristolochia tagala Champ. was extracted by methanol and its extract was fractionated into several solvent fractions. The chloroform fraction, which showed the highest antimicrobial activity, was separated by column chromatography and obtained forty three subfractions. The forty three fractions were searched the anti-fungal activities by bioassay. The most active No. 26 subfraction was analyzed by GC-MS. Each mass spectra, corresponding to each peak of chromatogram, was compared to MS database of Wiley library. As a result, 2,4-di-tetra-butyl-phenol, 2-mono-palmitin, 1-mono-stearin were profiled as maine compounds in No. 26 subfraction. Bioassay using commercial 1-mono-stearin to test for the anti-microbial activity conformed the antimicrobial active compound. In conclusion, 1-mono-stearin identified from Aristolochia tagala Champ. was antimicrobial chemical against Sclerotinia sclerotiorum.