초록 ▼

○ 연구결과
1) 육계의 급이 방법에 따른 유용 유전자 발현 양상 구명과 실용 표지 유전자 발굴; 지방 및 근육 발달 관련 유전자(PPARγ, FAS, MyoD 등) 클로닝, 노화 표지 개발(telomere), 사료 및 에너지 제한 급이는 지방 및 근육 관련유전자 발현에 영향 없음, 생산성에 부정적인 영향 없이 면역력 증강 및 생리활성도 증가
2) 육계의 생리활성물질 첨가 급여에 따른 유용 유전자 발현 양상 구명; 가시오갈피 및 두충 첨가 급여시 근육 관련 유전자의 발현 증대, 항산화 효소(SOD, GPX)의 활성 증진,

○ 연구결과
1) 육계의 급이 방법에 따른 유용 유전자 발현 양상 구명과 실용 표지 유전자 발굴; 지방 및 근육 발달 관련 유전자(PPARγ, FAS, MyoD 등) 클로닝, 노화 표지 개발(telomere), 사료 및 에너지 제한 급이는 지방 및 근육 관련유전자 발현에 영향 없음, 생산성에 부정적인 영향 없이 면역력 증강 및 생리활성도 증가
2) 육계의 생리활성물질 첨가 급여에 따른 유용 유전자 발현 양상 구명; 가시오갈피 및 두충 첨가 급여시 근육 관련 유전자의 발현 증대, 항산화 효소(SOD, GPX)의 활성 증진, 흉선에서 면역 사이토카인의 발현 증대 및 친염증 사이토카인의 발현 억제, 생리활성 증진 및 노화 억제에 긍정적 효과 발현
3) 육계의 사육 밀도에 따른 유용 유전자 발현 양상 구명; 지방 및 근육관련 유전자(FAS, PPARγ)의 발현은 사육밀도에 영향 받지 않음, 스트레스관련 유전자(HSP 70과 HMGCR) 발굴, 고밀도 사육이 친염증 사이토카인 발현을 증가시켜 면역력 저하를 발생, 텔로미어 함유율에 따라 사육밀도가 개체의 스트레스 요인 중 하나임을 입증
4) 산란계에서 천연 항산화 사료첨가제 개발; 가시오갈피 및 두충 급여가 항산화 방어기능에 효과적 영향, 계란의 난질 개선 효과, 백혈구 내 텔로미어 함유율이 현저히 높은 양상을 보임에 따라 생리 활성도 및 기능 향상에 영향

Abstract ▼

Ⅳ. Results
1. Effect of feed and energy restriction.
1) Effect of Feed restriction and Energy restriction on adipogenic and myogenic gene expression levels.
The expression of adipogenic genes like malic enzyme, FAS, SREBP1c, PPAR gamma, and C/EBP alpha were down regulated by restricted feed

Ⅳ. Results
1. Effect of feed and energy restriction.
1) Effect of Feed restriction and Energy restriction on adipogenic and myogenic gene expression levels.
The expression of adipogenic genes like malic enzyme, FAS, SREBP1c, PPAR gamma, and C/EBP alpha were down regulated by restricted feed regimen. Muscle related genes like MyoD, Myf5, and Myogenin were also down regulated at the end of d14, but the analysis at the end of the experiment at d35 the expression levels of malic enzyme and FAS did not show any difference when compared to the control group. Except ER85 (Energy restriction 85%), the expression levels of MyoD and Myf5 in all treatment group was decreased by restriction regimen compared to control group. The expression levels of Myogenin were down regulated by restriction regimen in all treatment groups. Myf5 gene from ER85 was highly expressed at d35 of age, but it was not different among other treatments. The RNA expression levels of MyoD and Myogenin were not different among treatment groups after realimenation at d35 of age in broilers. These results suggest that ER85 group shows the best growth performance by realimenation and the genes of muscle are also expressed as much as that of control group even during the restriction period. However, the ER70 group could not catch up the body weight with the control group and the FAS expression is still low at the end of d35 of age. ER70 is severe to normal growth of broilers and its better to avoid the restriction regimen in broiler chickens.
2) Effect of Feed restriction and Energy restriction on immune relateted gene expression levels.
There was no statistical difference in weight gain, feed intake and feed conversion ration between the CON and quatitative or qualitative FR groups during the entire periodis. In cytokines, there was no significant difference in the expression of lymphocytic IL-4 and IFN-gamma at 14d of age between the FR and the CON groups, whersas lymphocytic IL-6 and iNOS expression were significantly lower in FI85 and FI70 groups. In conclusion, 85% of quatitative or qualitative FR would have benefical effect on the expression of some cytokine without change in growth performance of birds.
3) Feed restriction and telomere
In here, we studied the effect of feed restriction on telomere amount and telomerase activity. Telomere amount is an age dependent biomarker as it is known to reduce with age. We investigated blood, liver, lung, heart, testes tissues in this study which involved three sampling at the end of 7 days, 14 days and 35 days respectively. In results, we found out that telomere amount decreased with age and hence proves its role as a age dependent biomarker. At the of 14 days, we found that the amount of telomere was decreased compared to controls however, at the end of the study after 35 days we found that telomere quantity was more than that of the control. The explanation for this change is probably because of the compensation effect on the study birds due to feed restriction. The difference in telomere amount was significant in case of blood lymphocytes, lung and testes tissues. The telomerase activity however, remained unchanged at end of 14 and 35 days. It showed positive activity only in case of testes tissue which is the gonadal tissue and hence needs telomerase activity for the normal telomere maintenance needed for the gametes.
2. Effect of natural antioxidants (Acanthopanax senticosus and Eucommiaceae)
1) Effect of dietary supplementation of siberian ginseng and Eucommiaceae on the expression of lipogenic, myogenic, and antioxidant enzyme genes in broiler chickens.
Birds subjected to 1% A. senticosus had higher feed conversion ration than the other treated birds where as no significant difference were found in body weight, weight gain and feed intake. The expression levels of FAS were not different among the treated groups while the transcription factors PPAR gamma was highly expressed in Eucommiaceae but not in control and A. senticosus. The gene expression of Myogenin was high in both the treatments compared to control. MyoD also showed high expression in treated groups furthermore, Eucommiaceae could stimulate the expression of MyoD more than A. senticosus. The antioxidant gene expression (SOD, CAT GPX) generally were not much different among the treatments, however GPX and SOD were stimulated in broilers consumed 1% Eucommiaceae diet. The result of this experiment showed that the dietary supplimentation of A.senticosus and Eucommiaceae in broiler may improve the antioxidant defence system through SOD and GPX without affecting the growth performance.
2) Effect of dietary supplementation of natural bioactive substances on growth performance, expression of genes associated with immune system in broiler chickens.
The blood biochemical profiles like albumin, AST, ALT and total protein were not different among treated groups, however the triglycerides (TG), cholesterol and glucose in treated groups significantly (P<0.05) increased compared to that of control group. The expression levels of cytokines (IFN gamma, IL-1beta, IL-10 and IL-4) of blood were not different among the treated groups, however the expression levels in thymus were significantly (P<0.05) higher in treated group compared to the control. The expression levels of TNF alpha, iNOS, IL-6 and IL-10 in liver were significantly lower in eucomiaceae supplemented birds.
3) Antioxidant and telomere
In this study we sought out to investigate the effects of antioxidant feed on broiler chicken telomere length in blood lymphocyte, liver, lung, heart and testes tissues. Since, telomere amount is an indication of the biological well being of a cell of an organism. The birds were fed with two diets consisting of Euchomia and Acantoponox senticosum. In results, we found out that antioxidant fed chicken showed higher telomere amounts than the control ones especially in blood and specifically Acantoponox senticosum feed effect was much more effective than the Euchomia fed birds. The telomere amount also reflects the immune state of the birds since it is involved in the well being of lymphocytes which are key players in immune function. Other tissues such as liver, lung, heart and testes showed little effect with respect to change in telomere amounts when compared to blood lymphocytes.
4) Effect of antioxidants in laying hens
All the birds were subjected t o 5 different treatments from the age 20 weeks to 30 weeks. Growth performance and physiological markers were analyzed. The expression levels of SOD showed no difference in intestine and liver tissues, but GST had an increased expression levels. Thus these antioxidants have immuno protective effect. Apart from this the quality of the egg in the treatment group was better than the control. As these natural antioxidants do have side effect so can be used as a feed supplement.
3. Effect of stocking density
1) Effect of stocking density on adipogenic, myogenic and stress gene expression levels.
We investigated the effect of stocking density on adipogenic (PPARgamma, Fattyacidsynthase), myogenic (Myogenin, Myo-D, Myf5), stress related heat shock protein genes (HSP25, HSP70 and HSP90), 3-hydroxil-3-methyl-glutaryl coenzyme A reductase (HMGCR) and apoptosis related (Bcl-X and Bcl-2) genes in broiler chickens. Except the feed intake (P<0.05), the other growth performances like feed conversion ratio, body weight gain, feed efficiency ratio showed no difference among the treated groups. HSP90, HSP25, PPAR gamma, FAS, Myogenin, Myo-D, Myf5, Bcl-x, and Bcl-2 did not show any significant changes in the expression among the treated groups. While HSP70 and HMGCR gene expression levels showed the increasing trend with the increase in stocking density. Though the expression levels were not significantly different among the low and standard group but were found to be significantly (p<0.05) high in high density stocked group. This clearly indicates that birds stocked at high density were experiencing stress when analyzed at transcription level.
2) Effect of stocking density on growth performance, expression of genes associated with immune system in broiler chickens.
Stocking density showed no significant effect on body weight and feed conversion ratio among the different treatments. However, feed intake was significantly (P<0.05) high in the low density treatment. The weights of liver, spleen and thymus were not different among the treatments. The expression levels of cytokines, iNOS, IFN gamma, IL-1, IL-4, IL-6 and TNF alpha were not different among the treatment, but the expression levels of IL-10 was significantly (P<0.05) enhanced in high density treatment when blood, bursa, spleen and liver were analyzed. However the cytokines, iNOS, IFN gamma, IL-1, IL-4, IL-6 and TNF alpha were significantly enhanced in thymus for high density treatment. The expression of IL-4 decreased in low density treatment. In conclusion, high density has very little effect on the immune function.
3) Stocking density and telomere
In this study, we investigated the effect of stocking density on telomere amount in blood, liver, lung, heart and testes tissues. We hypothesized that increased stocking density in broiler chickens causes increase in stress in the birds and hence would affect the amount of telomere amount in these birds. Using the interphase Q-FISH technique we analyzed the telomeric amount and In results, we found out that with increase in stocking density the amount of telomere decreases in blood lymphocytes however, we did not find much change in the other tissues

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 10
• CONTENTS ... 16
• 목차 ... 17
• 제 1 장 연구개발과제의 개요 ... 18
• 제1절 연구개발의 목적 ... 18
• 제2절 연구개발 필요성 ... 18
• 제3절 연구개발의 내용 및 범위 ... 19
• 제 2 장 국내외 기술개발 현황 ... 22
• 1. 세계적 수준 ... 22
• 2. 국내수준 ... 22
• 3. 국내․외의 연구현황 ... 22
• 제 3 장 연구개발수행 내용 및 결과 ... 23
• 제1절 육계의 급이 방법에 따른 유용유전자 발현양상 구명과 실용표지유전자 발굴 ... 23
• 제2절 생리활성 물질 첨가 급여에 따른 유용유전자 발현양상 구명과 실용 표지유전자 발굴 ... 50
• 제3절 육계의 사육 밀도에 따른 유용유전자 발현양상 구명과 실용 표지유전자 발굴 ... 74
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 92
• 제1절 목표달성도 ... 92
• 제2절 관련분야 기여도 ... 95
• 제 5 장 연구개발 성과 및 성과활용 계획 ... 96
• 제1절 연구 성과 ... 96
• 제2절 성과활용 계획 ... 100
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 101
• 제 7 장 참고문헌 ... 102
• 끝페이지 ... 107