초록 ▼

○ 연구결과 :
1. 반추위미생물의 hydrogenation activity 조절에 관한 연구
1) 일반 사료자원의 탄수화물성 에너지 수준 : 에너지 수준이 높을수록 pH가 낮아 CLA 생성률이 저하됨
2) 섬유소(일반 조사료) 첨가 수준 : 섬유소 첨가수준이 CLA 생성에 크게 영향하지 않았음
3) 지질자원의 첨가형태 : oil 자원의 종류 및 oil 또는 oilseed의 첨가 형태에 따라 CLA 생성률이 다르게 나타났음
4) 배양액의 pH : 배양액의 pH가 높을수록 CLA 생성량이 증가되었음
2

○ 연구결과 :
1. 반추위미생물의 hydrogenation activity 조절에 관한 연구
1) 일반 사료자원의 탄수화물성 에너지 수준 : 에너지 수준이 높을수록 pH가 낮아 CLA 생성률이 저하됨
2) 섬유소(일반 조사료) 첨가 수준 : 섬유소 첨가수준이 CLA 생성에 크게 영향하지 않았음
3) 지질자원의 첨가형태 : oil 자원의 종류 및 oil 또는 oilseed의 첨가 형태에 따라 CLA 생성률이 다르게 나타났음
4) 배양액의 pH : 배양액의 pH가 높을수록 CLA 생성량이 증가되었음
2. 반추위내 기능성지방산의 생산 증가를 위한 Ionophore 첨가효과 구명
1) oil 배양시 Monensin 첨가로 인하여 CLA 생성량이 증가되었음
2) oil의 종류에 따른 불포화지방산의 hydrogenation 정도가 달랐음
3. Batch culture와 continuous culture를 이용한 지방 첨가가 반추위 미생물의 조성에 미치는 영향
1) oil 종류와 미생물에 따라 지방산의 농도와 조성이 달랐음
2) 반추위미생물에 의한 biohydrogenation비율이 C18:3> C18:2> C18:1 순서로 나타났음
3) continuous culture 방법의 경우 oil 첨가로 인하여 미생물 단백질 함량이 다소 낮아졌으며, 미생물의 지방산 조성이 oil의 종류에 따른 지방산 조성의 영향을 받았음
4. 지질사료의 급여조건에 따른 불포화지방산의 hydrogenation 및 T-FA 생성과 이들 물질의 체내 이용성에 관한 연구
1) 조사료 비율이 증가됨에 따라 두 종류의 oil 모두에서 trans11-ODA와 cis9, trans11-CLA 함량이 증가되는 경향을 보였음
2) 그러나 농후사료와 조사료 비율 그리고 oil의 종류는 plasma의 trans11-ODA와 cis9, trans11-CLA 함량에 크게 영향하지 않았음
3) 대두유의 첨가는 면양의 plasma 내 total cholesterol 및 HDL-cholesterol 함량을 증가시켰음
4) 대두유의 첨가로 인하여 근내지방과 피하지방의 CLA 함량이 다소 증가되었음
5. Ionophore가 반추동물의 기능성지방산 생산에 미치는 효과
1) 식물성 기름이나 Monensin의 반추위 내 투여로 장쇄 지방산의 반추위 내 hydrogenation을 감소 시켰음
2) 대두유와 Monensin 투여로 인하여 반추위 내용물의 CLA 함량이 크게 증가되었음
6. 한국재래산양 대사시험을 통한 지방첨가가 반추위 내 대사에 미치는 영향 조사
1) 반추위미생물과 혈액 내에 존재하는 C18계 unsaturated fatty acid는 지방 첨가구에서 hydrogenation이 높았음
2) Oil의 첨가로 인하여 반추위 미생물의 성장이 둔화되었음
7. 지질의 첨가가 체지방의 T-FA 및 지방산 조성에 미치는 효과
1) 대두유의 첨가는 면양의 plasma 내 total cholesterol 및 HDL-cholesterol 함량을 증가시켰음
2) 대두유의 첨가로 인하여 근내지방과 피하지방의 CLA 함량이 다소 증가되었음
8. 지질사료의 최적 급여조건을 통한 기능성 한우고기의 생산
1) 5개월간에 걸친 oil 혼합물 + Monensin(MOPM)의 급여로 인하여 일당증체량과 사료효율이 현저히 증가되었음
2) MOPM 사료는 거세 한우의 등지방 두께와 등심근 면적을 다소 증가시켰음
3) MOPM 첨가사료 급여로 plasma의 cholesterol 농도와 HDL-C 농도 그리고 HDLC/TC 비율을 증가시켰음
4) MOPM 첨가사료 급여는 거세 한우의 plasma c9,t11-CLA 함량을 현저히 증가시켰음
5) 5개월간에 걸친 MOPM 첨가사료 급여로 인하여 근내지방, 근간지방 및 피하지방의 CLA 함량과 총 불포화지방산 함량이 증가되었으며 포화지방산 함량은 감소되었음

Abstract ▼

1) Effect of energy level on the formation of trans-C_18:1 acid and the degree of hydrogenation from cis-unsaturated fatty acids by rumen microbes
An in vitro study was conducted to examine the effect of addition level of concentrate on fermentation characteristics and long-chain unsatu

1) Effect of energy level on the formation of trans-C_18:1 acid and the degree of hydrogenation from cis-unsaturated fatty acids by rumen microbes
An in vitro study was conducted to examine the effect of addition level of concentrate on fermentation characteristics and long-chain unsaturated fatty acids composition, especially conjugated linoleic acid(CLA) and trans-octadecenoic acid(t-FA) by mixed ruminal bacteria when incubated with linseed or rapeseed. Four levels(0.83, 1.25, 1.67 and 2.08%, w/v) of concentrate and ground oilseeds(linseed or rapeseed; 0.83%, w/v) were added to mixed solution of strained rumen fluid with artificial saliva(1:1, v/v) in the glass jar with a glass lid equipped with stirrer, and was incubated anaerobically for 24 hours at 39℃. Addition level of concentrate slightly reflect on pH and ammonia concentration of the culture solution at the various incubation times when incubated with both linseed and rapeseed. Total VFA concentration slightly increased with incubation times and concentrate levels for incubations with oilseeds. Percent CLA had a clearly decreasing trend with concentrate level throughout incubation times with significances at 24h incubations when incubated with rapeseed(P<0.031). The t-FA/CLA ratios was lower for rapeseed with increased proportion of CLA and decreased t-FA proportion than for linseed.
2) Effect of roughage level on on the formation of trans-C_18:1 acid and the degree of hydrogenation from cis-unsaturated fatty acids by rumen microbes
An in vitro study was conducted to examine the effect of different level of alfalfa hay on fermentation characteristics and long-chain unsaturated fatty acids composition, especially conjugated linoleic acid(CLA) and trans-octadecenoic acid(t-FA) with rapeseed supplementation by mixed ruminal bacteria. Three levels alfalfa hay were added to 600 ml ruminal culture at the level of 0.25, 0.50 and 0.75%(w/v), respectively. A semi-continuous ruminal fermentation system was used in the present study. Addition of alfalfa hay to the culture at different levels did not affect significantly the pH, ammonia concentration and volatile fatty acid(VFA) concentration in ruminal culture. Bacteria growth in culture solution had a decreasing trend with alfalfa hay level at 12h incubation. The clear increased trends on CLA proportion, while decreased trends on t-FA/CLA ratio were found from the treatments containing 0.50 and 0.75% alfalfa addition than those control and 0.25% alfalfa addition which were similar at 3h incubation. There were decrease trends in C18:0 proportion, while the proportion of C18:2 and C18:3 tended to increase with alfalfa hay level at 12h and 24h incubation.
3) Effect of pH in incubation solution on on the formation of trans-C_18:1 acid and the degree of hydrogenation from cis-unsaturated fatty acids by rumen microbes
An effect of pH on the fermentation characteristics and the formation of cis-9,trans-11 conjugated linoleic acid(CLA) and trans-11 octadecenoic acid (t-FA) by mixed ruminal bacteria was examined in vitro when incubated with linseed or rapeseed. Concentrate(1%, w/v) with ground linseed(0.6%, w/v) or rapeseed(0.5%, w/v) was added to 600 ml mixed solution of strained rumen fluid with artificial saliva(1:1, v/v), and was incubated anaerobically for 12 hours at 39℃. The pH of culture solution was maintained at close levels of 4.5, 5.3, 6.1 and 6.9 with 30% of H2SO4 or 30% NaOH solution.
pH increment resulted in increases of ammonia and total VFA concentration in culture solutions containing both oilseeds. Fermentation was not proceeded at pH 4.5. Molar proportion of acetate(C2) decreased but that of propionate(C3) increased as pH increased when incubated with oilseeds. While hydrogenating process was very slow at the pH range of 4.5 to 5.3, rapid hydrogenation was found from the culture solutions of pH 6.1 and 6.9 when incubated with linseed or rapeseed. As pH in culture solution of linseed or rapeseed increases proportions of oleic acid and t-FA increased but those of linoleic acid and linolenic acid decreased. The CLA proportion increased with pH in culture solution containing rapeseed but CLA was not mostly detected from the incubation of linseed.
4) Effect of addition type of oil source the formation of trans-C_18:1 acid and the degree of hydrogenation from cis-unsaturated fatty acids by rumen microbes
The effects of addition type of oil source on fermentation characteristics and long-chain unsaturated fatty acids composition, especially the formation of conjugated linoleic acid(CLA) and octadecenoic acid(trans-11 C_18:1, t-C_18:1) by mixed ruminal bacteria were examined in vitro. Concentrate (1% of culture solution, w/v, as-fed basis) with ground linseed(0.6% of culture solution, w/v, DM basis) or linseed oil as absorbed into the ground alfalfa hay were added to 600 ml mixed solution consisting of strained rumen fluid and artificial saliva at the ratio of 1:1 in the glass culture jar. The culture jar was covered with a glass lid equipped with stirrer and was placed into a water-bath(39℃) and was incubated anaerobically up to 24hrs. Addition type of oil source did not affect the pH and ammonia concentration. Decreasing trends in the compositions of C18:2 and C18:3 but increasing trends of C18:0 compositions were clearly found from culture contents up to 12h incubation when incubated with both ground rapeseed and RSO(Table 25).
The effects of oil addition type on FA composition in culture solution were clear from 3h incubation. The compositions of C_18:0, C_18:1 and C_18:2 were greater(P<0.0001∼.033) but CLA were smaller (P<0.0008∼.0032) in a culture solution containing RSO than those in a culture solution of ground rapeseed.
5) Studies on the effect of Ionophore on CLA production by rumen microbes
An incubation with rumen fluid was conducted in a 2×2 factorial arrangement of treatments. Treatments were 1) control, 2) 0.80ppm monensin, 3) 5% soybean oil, and 4) 0.8 ppm monensin + 5% soybean oil which were added to rumen fluid and incubated for 0, 3, 6 and 9h.
Monensin and soybean oil treatments decreased acetate to propionate ratio significantly(P<.05) without changing total VFA concentrations. Monensin and soybean oil treatements also increased accumulation of cis C_18:1, trans C_18:1,and CLA but decreased that of C18:0 in incubation contents. Significant (P<.05) decreases in C_18:3 and C_18:2 were observed in the control, whereas the monensin treatment showed delayed disappearance of C_18:3 and C_18:2, as incubation time proceeded. Supplementation of both monensin and soybean oil increased the concentrations of trans C_18:1 and CLA but decreased C_18:0 significantly(P<.05) in the rumen contents when compared to other treatments. Yield of CLA in rumen contents was also significantly(P<.05) higher for the monensin + soybean oil treatment among all treatments.
6) Effect of oil addition on fatty acid composition of rumen microbes by batch culture
The present study was to determine the fatty acid composition of four common rumen bacteria species and mixed culture with additional oil sources on ruminal fermentation in vitro. pH was optimally maintained but pure culture was lower then additional oil sources. NH3-N concentration was important factor synthesis of microbial protein synthesis. However, this result was related to microbial protein synthesis in ruminal culture. Additional oil sources with mixed culture were lower NH3-N concentration. Additional oil sources decreased concentration of total VFA according as acetate content in mediums during incubation time, but mixed culture was similar concentration of total VFA. A/P ratio was shown amount of propionic acid lower then decreased amount of acetic acid of four common rumen bacteria
species.
Microbial dry matter was incubated in additional oil sources lower than pure culture and mixed substrate. Especially, linseed oil treatments of all cultures were more little microbial dry matter than CSPO treatment. The effect of rumen microbes in a 24 h incubation showed that the linseed oil was biohydrogenation trans-C(41.97%), cis-C18:1(54.97%), C18:2(68.96%) and C18:3(50.78%).
Although biohydrogenation of CSPO was significantly different, the rumen microbes hydrogenated the trans-C18:1(31.58%), cis-C18:1(45.45%) and C_18:2 (59.92%), C18:3(66.63%), there was a corresponding increase in C18:0. C18:0 of linseed oil was higher than CSPO, which effect of CSPO was protected hydrogenation by rumen microbes and higher value of C18 UFA. Biohydrogenation percent had the highest followed by C18:3>C18:2>C181. The end-products of C18 unsaturated fatty acid by rumen microbes were stearic acid and trans-octadecanoic isomers, which is an associated with effects of ruminal lipolysis and hydrogenation.
7) Effect of oil addition on fatty acid composition of rumen microbes by continuous culture
An experiment with a 4×4 Latin square design was conducted to determine the effects of supplemental dietary fat on rumen microbial fermentation in continuous culture. pH was optimally maintained but pure culture was lower then additional oil sources and sunflower oil treatment high pH of oil source treatment. NH3-N concentration was not significant different lower at control than oil source treatments. Microbial protein synthesis was not significant different higher control than oil source treatment. However, this result was related to microbial protein synthesis in ruminal culture. Additional soybean oil treatment and control increased concentration of total VFA(P<0.05). A/P ratio was significantly higher control than oil source treatments(P<0.05). Unsaturated fatty acid concentration of oil source treatments was highest in sunflower oil treatment. C18:0, C18:1 and C18:3 of fatty acid composition of bacteria in suspension fluid and vessel content was not significantly different high on sunflower oil treatment. Biohydrogenation ratio not significantly different was highest sunflower oil treatment.
2. Metabolism studies
1) Effects of concentrate to roughage ratio on the lipid metabolism in rumen and fatty acid composition of plasma in sheep
A metabolism trial with four ruminally fistulated sheep was conducted in a 4 x 4 Latin square design to examine the effect of concentrate to roughage ratio(70:30 vs. 85:15) and oil source(soybean oil vs. rapeseed oil) on the ruminal fermentation pattern and C₁₈-fatty acids composition including trans11-C18:1(trans11-ODA) and cis9, trans11-C_18:2(cis9, trans11 -CLA) in the rumen fluid and plasma. Oil was added to the concentrate at 5% level of the total diet(DM basis) and chopped rye grass hay was fed as roughage.
An increased level of concentrate(85%) within supplemented oil slightly lowered pH but increased ammonia concentration. Supplementation of rapeseed oil relatively increased pH and ammonia concentration. Higher concentrate level resulted in increased tendencies of total VFA concentration while oil source did not affect the total VFA concentration and VFA proportion. Whole tract digestibilities of DM, CP, EE, NDF and OM in diets slightly increased at higher concentrate level.
Proportions of oleic acid(C_18:1) and linoleic acid(C_18:2) in the rumen fluid were influenced by the fatty acid composition of oil source but oil source did not affect the in vitro formations of trans11-ODA and cis9, trans11-CLA. Slightly increased trans11-ODA and cis9, trans11-CLA proportions, however, were observed from the sheep fed high roughage diet supplemented with both soybean oil and rapeseed oil.
The C18:1 and C18:2 composition in supplemented oils responded to those in plasma of sheep. Effects of concentrate to roughage ratio and oil source on trans11-ODA and cis9, trans11-CLA proportions in plasma were found to be small. Proportion of cis9, trans11-CLA in plasma tended to be increased from the sheep fed high roughage diet and collection time at 9h post feeding.
2) Effect of Ionophore on the procuction of CLA in the rumen of ruminants
Five ruminally cannulated rams were used in a 5x5 Latin square design experiment to study the effects of plant oil added with or without monensin on the changes of fermentation pattern and long chain fatty acids including CLA. The treatments were 1) Control, 2) Corn oil, 3) Corn Oil+Monensin, 4) Soybean Oil, and 5) Soybean Oil+Monensin. The animals were fed 60 part grass hay and 40 part mixed concentrates. and plant oils were given in a form adsorbed in concentrate pellet. Monensin-Na was given at 33ppm through the rumen cannula.
Feeding plant oil or monensin significantly(P<.05) inhibited ruminal biohydrogenation of long chain fatty acids fed with concentrate feed. Soybean oil + monensin treatment showed the highest yield of CLA in the rumen content. Even if corn oil showed less yield of CLA in the rumen, formed higher concentration of trans-C18:1 could later used within the tissue being used as precurser for an endogenous synthesis of CLA.
It was concluded from the results of in vitro and in vivo experiments that monensin fed with plant oils increased CLA accumulation in the rumen by inhibiting complete biohydrogenation of respective C_18:3 and C_18:2 to C_18:0.
Supplementation of both soybean oil and monensin in a diet will be a useful feeding practice to increasing CLA contents in ruminants' products.
3) Effect of oil supplementation on lipid metabolism in Korean native goat
Metabolism study was conducted with four ruminally cannulated Korean native goats to examine the effects of oil supplementation on fermentation characteristics, fatty acid composition of rumen microbes and blood by 4 x 4 Latin Square design.
Basal diet consisted of 60% concentrate and 40% rice straw(DM basis).
Experimental diets were basal diet as a control, supplementations of linseed oil, soybean oil, and sunflower oil at 5% level(DM basis) of basal diet.
Concentration of C18:2 in rumen microbes was significantly highest(P<0.05) for sunflower oil treatment, but concentrations of C18:1 and C18:3 were lower than those of control. This results similar to blood fatty acid composition and unsaturated fatty acid was higher than control. Biohydrogenatin of unsaturated fatty acid on rumen and blood tend to increased in oil source treatment.
3. Feeding studies
1) Effect of soybean oil supplementation on trans-11 octadecenoic acid and cis9,trans11-CLA content in the tissues of sheep
A feed trial was conducted with sheep for 12weeks to examine the effect of soybean oil(SBO) supplementation on long-chain fatty acids composition, especially trans-11 octadecenoic(t-FA) and cis9,trans11-CLA (c9,t11-CLA) in various tissues.
Sheep were fed either a SBO supplemented diet(5%, DM basis) or a control diet without SBO. Chopped rye grass hay was fed as roughage. Concomitant increases in contents of TC and HDLC in the plasma of sheep were observed from the SBO supplementation, resulting in the increased ratio of HDLC/TC. The supplementation of SBO reduced(P<0.05) the proportions of C16:1, C17:0 and C17:1 but increased (P<0.05) the proportions of C18:0 and total t-FA in the intramuscular fat.
The C18:0 proportion only in the subcutaneous fat was increased(P<0.05) by the SBO supplementation. Proportions of C_15:0 and C_18:0 in the kidney fat were also increased(P<0.05) while those of C_16:1 and t-FA were reduced (P<0.05) by the SBO supplementation. The SBO supplementation slightly increased CLA proportion in the intramuscular fat and subcutaneous fat.
2) Production of functional Hanwoo beef by the optimal feeding of dietary lipid
Twenty-four Hanwoo steers(20 months old, 567±18kg body weight) were divided into three groups of eight animals each according to body weight.
Hanwoo steers in control group had the commercial concentrate for the late fattening stage. The other groups of steers were fed the same diet as control steers, but the concentrate was supplemented with high-C18:2 oil mixture(45% SBO, 20% sunflower oil, 20% safflower oil and 15% fish oil) at 6% level of concentrate(DM basis) and monensin(20ppm). The second and third group of steers were fed the oil mixture supplemented diet with monensin for last 2.5months and 5months, respectively, before they were being slaughtered.
Average daily gain(ADG) of Hanwoo steers during the 5month of experiment was higher(P<0.049) for 5 month feeding of diet supplemented with oil mixture and monensin(MOPM) than for the other groups of steers, thus improved feed efficiency was observed from 5month feeding group. No difference in ADG was
found between control and 2.5month feeding group of steers.
Fat thickness and longissimus muscle area tended to be greater in steers fed the diet supplemented with MOPM than in control steers. The MOPM supplementation for 2.5 months increased the pH(P<0.006), decreased the CIE a* value(P<0.04) compared to control. Feeding the MOPM supplemented diet for 5month improved quality grade while lowered the quantity grade of Hanwoo steers compared to control.
The MOPM supplementation increased total cholesterol(TC, P<0.0001∼.0007), HDL-C (P<0.0001) and HDLC/TC(P<0.004∼.034) ratio in plasma after steers were fed for 3 month and 5 month compared with control steers. Steers fed MOPM had approximately twice the level of the c9,t11-CLA in plasma(0.84% vs. 0.44%) than the control steers.
Supplementation of MOPM for 5 month increased the content of CLA in intramuscular(P<0.015), intermuscular(P<0.039) and subcutaneous(P<0.001) fat tissues compared with those of steers fed control diets and diets supplemented for 2.5 months which were similar, respectively. Total UFA was increased, while SFA was decreased(P<0.008) due to the increased(P<0.001) C18:2 and decreased(P<0.002) C18:0 proportions in subcutaneous tissue when steers were fed diets supplemented with MOPM for 5 month compared with those of steers fed control diets and diets supplemented for 2.5 months which were similar.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 2
• SUMMARY ... 13
• CONTENTS ... 21
• 목차 ... 23
• 제1장 연구개발과제의 개요 ... 24
• 제1절 연구개발의 목적 및 필요성 ... 24
• 1. 기술적 측면 ... 24
• 2. 경제·산업적 측면 ... 25
• 3. 사회·문화적 측면 ... 25
• 제2절 연구개발의 목표 및 내용 ... 26
• 제2장 국내외 기술개발 현황 ... 28
• 제3장 연구개발수행 내용 및 결과 ... 35
• 제1절 In vitro 시험 결과 ... 35
• 1. 연구 제목 : 반추위미생물의 Hydrogenation Activity 조절에 관한 연구 (충북대) ... 35
• 2. 연구 제목 : 반추위 내 기능성지방산의 생산 증가를 위한 Ionophore 첨가효과 구명 (고려대) ... 62
• 3. 연구 제목 : 사료 내 지방첨가에 따른 반추위 내 미생물 대사에 미치는 영향 (중앙대) ... 83
• 제2절 대사시험 결과 ... 100
• 1. 연구 제목 : 지질사료의 급여조건에 따른 불포화지방산의 hydrogenation 및 T-FA 생성과 이들 물질의 체내 이용성에 관한 연구 (충북대) ... 100
• 2. 연구 제목 : Ionophore가 반추동물의 제1위 내 CLA 생산에 미치는 효과 연구 (고려대) ... 108
• 3. 연구 제목 : 한국재래산양 대사실험을 통한 지방 첨가가 반추위 내 대사에 미치는 영향 (중앙대) ... 119
• 제3절 사양시험 결과 ... 128
• 1. 연구 제목 : 지질의 첨가가 체지방의 T-FA 및 지방산 조성에 미치는 효과 (충북대) ... 128
• 2. 연구 제목 : 지질사료의 최적 급여조건을 통한 기능성 한우고기의 생산 (충북대) ... 133
• 제4절 결과 종합 ... 145
• 제4장 목표달성도 및 관련분야에의 기여도 ... 149
• 제5장 연구개발결과의 활용계획 ... 153
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 155
• 제7장 참고문헌 ... 156
• 끝페이지 ... 170