• 제목/요약/키워드: Replication protein A

검색결과 328건 처리시간 0.022초

Effects of dietary leucine supplementation on the hepatic mitochondrial biogenesis and energy metabolism in normal birth weight and intrauterine growth-retarded weanling piglets

  • Su, Weipeng;Xu, Wen;Zhang, Hao;Ying, Zhixiong;Zhou, Le;Zhang, Lili;Wang, Tian
    • Nutrition Research and Practice
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    • 제11권2호
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    • pp.121-129
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    • 2017
  • BACKGROUND/OBJECTIVES: The study was conducted to evaluate the effects of dietary leucine supplementation on mitochondrial biogenesis and energy metabolism in the liver of normal birth weight (NBW) and intrauterine growth-retarded (IUGR) weanling piglets. MATERIALS/METHODS: A total of sixteen pairs of NBW and IUGR piglets from sixteen sows were selected according to their birth weight. At postnatal day 14, all piglets were weaned and fed either a control diet or a leucine-supplemented diet for 21 d. Thereafter, a $2{\times}2$ factorial experimental design was used. Each treatment consisted of eight replications with one piglet per replication. RESULTS: Compared with NBW piglets, IUGR piglets had a decreased (P < 0.05) hepatic adenosine triphosphate (ATP) content. Also, IUGR piglets exhibited reductions (P < 0.05) in the activities of hepatic mitochondrial pyruvate dehydrogenase (PDH), citrate synthase (CS), ${\alpha}$-ketoglutarate dehydrogenase (${\alpha}$-KGDH), malate dehydrogenase (MDH), and complexes I and V, along with decreases (P < 0.05) in the concentration of mitochondrial DNA (mtDNA) and the protein expression of hepatic peroxisome proliferator-activated receptor-${\gamma}$ coactivator $1{\alpha}$ (PGC-$1{\alpha}$). Dietary leucine supplementation increased (P < 0.05) the content of ATP, and the activities of CS, ${\alpha}$-KGDH, MDH, and complex V in the liver of piglets. Furthermore, compared to those fed a control diet, piglets given a leucine-supplemented diet exhibited increases (P < 0.05) in the mtDNA content and in the mRNA expressions of sirtuin 1, PGC-$1{\alpha}$, nuclear respiratory factor 1, mitochondrial transcription factor A, and ATP synthase, $H^+$ transporting, mitochondrial F1 complex, ${\beta}$ polypeptide in liver. CONCLUSIONS: Dietary leucine supplementation may exert beneficial effects on mitochondrial biogenesis and energy metabolism in NBW and IUGR weanling piglets.

한탄바이러스의 섬유아세포 감염에 대한 Integrin 항체의 억제 효과 (The Suppressive Effects of Integrin Antibodies on the Infection of Hantaan Virus in Fibroblasts)

  • 박호선;김기득;김성광
    • Journal of Yeungnam Medical Science
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    • 제15권1호
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    • pp.55-66
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    • 1998
  • 사람과 동일한 증상을 나타내는 동물 모델이 없는 신증후출혈열의 병리 기전을 이해하기 위하여 in vitro culture system을 이용하였다. 신증후출혈열의 주된 병변이 혈관내피세포외에 섬유아세포가 많이 존재하는 신세뇨관 간질 및 폐간질임을 기초로 하여 계태아와 Mongolian gerbil의 섬유아세포에서 fibronectin에 대한 수용체인 ${\alpha}_5{\beta}_1$, integrin에 대한 항체를 처리하여 한탄바이러스 감염에 대한 영향을 관찰하였다. 초대 배양하여 사용한 계태아 섬유아세포 및 MGF 모두 한탄바이러스가 잘 감염되었으나 계태아 섬유아세포에 비하여 MGF에서 바이러스의 증식이 더 우수하였다. 계태아 섬유아세포와 MGF에서 ${\alpha}_5{\beta}_1$, integrin에 대한 항체 처리 5일 후 바이러스의 N 단백의 역가는 항체를 처리하지 않은 대조군에 비하여 각각 32.6%, 72.6%로 virion의 역가는 26.8%, 28.7%로 감소하여 두 세포에서 모두 감염력을 가진 바이러스의 양은 거의 동일하게 감소 되었다. 또한 ${\alpha}_5$ subunit와 ${\beta}_1$, subunit에 대한 항체를 MGF 세포에 처리한 후 바이러스의 N단백의 역가는 65.2%, 59.7%로 ${\alpha}_5{\beta}_1$ integrin에 대한 항체 처리하였을 때(72.6%)와 거의 유사하게 감소하였고 감염력을 가진 바이러스의 양도 26.5%, 29.4%로 감소하여 ${\alpha}_5{\beta}_1$ integrin에 대한 항체 처리시(28.7%)와 거의 유사하게 감소하였다. 이러한 결과는 ${\alpha}_5{\beta}_1$ integrin이 한탄바이러스에 대한 수용체로써 작용할 가능성을 시사하였으며 또다른 기전으로는 ${\alpha}_5{\beta}_1$ integrin의 차단에 의한 이차적인 영향으로 한탄바이러스의 증식에 커다란 영향을 미치는 것을 생각할 수 있었다.

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Study on Persistent Infection of Japanese Encephalitis Virus Beijing-l Strain in Serum-free Sf9 Cell Cultures

  • Kim, Hun;Lee, Su-Jeen;Park, Jin-Yong;Park, Yong-Wook;Kim, Hyun-Sung;Kang, Heui-Yun;Hur, Byung-Ki;Ryu, Yeon-Woo;Han, Sang-In
    • Journal of Microbiology
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    • 제42권1호
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    • pp.25-31
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    • 2004
  • Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

Comparison of the Effect of Green Tea By-product and Green Tea Probiotics on the Growth Performance, Meat Quality, and Immune Response of Finishing Pigs

  • Ko, S.Y.;Bae, I.H.;Yee, S.T.;Lee, S.S.;Uuganbayar, D.;Oh, J.I.;Yang, C.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권10호
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    • pp.1486-1494
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    • 2008
  • The objective of this experiment was to compare the effects of green tea by-product and green tea probiotics on the growth performance, meat quality and immune response of finishing pigs. A total of 72 crossbred "Landrace$\times$Yorkshire" finishing pigs with an average of 76 kg body weight were assigned to 4 dietary treatments in a completely randomized design. Each treatment had 3 replications with 6 pigs per replication. The four dietary treatments were control, antibiotics (control diet with 0.003% chlortetracycline added), and diets containing 0.5% green tea by-product or 0.5% green tea probiotic supplementation. Weight gain was increased in 0.5% green tea probiotics treatment compared to others, but there was no significant difference (p>0.05). The incorporation of 0.5% green tea probiotics to diets reduced the feed conversion ratio in finishing pigs (p>0.05). The incorporation of 0.5% green tea by-product into the pig diet reduced the crude protein and fat contents of the meat (p>0.05). Pigs fed diets containing 0.5% green tea probiotic supplementation had lowered meat TBA values compared to those fed 0.5% green tea by-product (p<0.05). The proliferation of spleen cells stimulated with Con A (concanavalin: 0.1, 0.3, and $1.0{\mu}g/ml$) significantly increased with 0.5% green tea by-product treatment compared to antibiotic treatment (p<0.05), but was significantly decreased in 0.5% green tea probiotics treatment compared to the antibiotic treatment (p<0.05). When stimulated with $1.0{\mu}g/ml$ Con A, splenocyte production of IL-6 from pigs treated with 0.5% green tea by-product or green tea probiotics was significantly increased compared to the antibiotic treatment group (p<0.05). Splenocyte production of TNF-${\alpha}$ after treatment with $1.0{\mu}g/ml$ Con A was significantly higher following 0.5% green tea probiotics treatment (p<0.05), while TNF-${\alpha}$ production after $10.0{\mu}g/ml$ LPS (lipopolysaccharide) was significantly higher in the 0.5% antibiotic treatment group (p<0.05).

Effect of Green Tea Probiotics on the Growth Performance, Meat Quality and Immune Response in Finishing Pigs

  • Ko, S.Y.;Yang, C.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권9호
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    • pp.1339-1347
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    • 2008
  • The objective of this study was to determine the effects of green tea probiotics on growth performance, meat quality and immune response in finishing pigs, and to assess the possibility of substituting green tea probiotics for antibiotics in diets of finishing pigs. This green tea probiotics is made by mixing green tea powder and excipients (defatted rice bran and wheat bran) and fermenting the mixture with beneficial bacteria. A total of 90 crossbreed "Landrace$\times$Yorkshire" finishing pigs with an average body weight of $72.5{\pm}2.5kg$ were assigned to 5 dietary treatments in a completely randomized design. Each treatment had 3 replications with 6 pigs per replication. The five dietary treatments were control, antibiotic (0.003% chlortetracycline added) and 0.1, 0.5 and 1.0% of green tea probiotics. There were no significant differences in final body weight, daily weight gain, daily feed intake and feed conversion ratio in the green tea probiotics and antibiotic treatments (p>0.05). Crude protein content was significantly increased in the 0.1 and 1.0% green tea probiotics treatment groups (p<0.05) and there was no significant difference in crude fat content of the meat among the treatments. The TBA value of meat was significantly lowered with 0.5 and 1.0% green tea probiotics treatments compared to that of controls and statistically similar to the antibiotic treatment after 3 weeks of storage (p<0.05). The growth of spleen cells stimulated with Con A (0.1 and $1.0{\mu}g/ml$) was significantly increased with 1.0% green tea probiotics treatment compared to that of the control treatment (p<0.05). The growth of spleen cells stimulated with LPS (1.0, 3.0 and $10{\mu}g/ml$) was significantly increased in the 0.5% green tea probiotics group compared to the antibiotic group (p<0.05). In Con A ($1.0{\mu}g/ml$) medium, IL-6 production of spleen cells was significantly increased with 1.0% green tea probiotics treatment compared to that of the control (p<0.05). In LPS ($10.0{\mu}g/ml$) medium, TNF-${\alpha}$ production of spleen cells increased significantly in all green tea probiotics treatment groups compared to that of the control (p<0.05). Finally it can be summarized that addition of green tea probiotic has a positive effect similar to antibiotic and 0.5% is the suitable dietary supplementation dose for finishing pig production.

The Uptake of 2-deoxy-D-glucose (2dGlc) by the Endogenous Sugar Transporter(s) of Spodoptera frugiperda Clone 21-AE Cells and the Inhibition of 2dGIc Transport in the Insect Cells by Fructose and Cytoc halasin B

  • Lee, Chong-Kee
    • 대한의생명과학회지
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    • 제9권4호
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    • pp.177-181
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    • 2003
  • The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

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Substitution of soybean meal with detoxified Jatropha curcas kernel meal: Effects on performance, nutrient utilization, and meat edibility of growing pigs

  • Li, Yang;Chen, Ling;Zhang, Yuhui;Wu, Jianmei;Lin, Yan;Fang, Zhengfeng;Che, Lianqiang;Xu, Shengyu;Wu, De
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권6호
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    • pp.888-898
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    • 2018
  • Objective: The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJM) in growing pig diets on growth performance, nutrients digestibility and meat edibility. Methods: A total of 144 pigs with initial body weight of $20.47{\pm}1.44kg$, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. Six diets (DJM0, DJM15, DJM30, DJM45, DJM60, and DJM75) were formulated using DJM to replace 0%, 15%, 30%, 45%, 60%, and 75% of SBM. From d 37 to 42, feces and urine were total collected from six barrows in each treatment. At day 79, thirty-six pigs were slaughtered for sampling. The feed intake and weight gain were recorded, while the intestinal morphology, digestive enzyme activities, nutrient digestibility and the content of residual phorbol esters in muscles were determined. Results: The results showed that increasing the replacement of SBM with DJM decreased the parameters including body weight, average daily gain, average daily feed intake, gain-to-feed ratio, weight and villus heights of duodenum, villus height and villus height/crypt depth of jejunum, digestive enzymes (protease, amylase, lipase, and trypsin) activities, and nutrients digestibility (nitrogen deposition, digestibility of nitrogen, energy digestibility, and total nitrogen utilization) (linear, p<0.05; quadratic, p<0.05) and there was no significant difference among DJM0, DJM15, and DJM30 in all measured indices. The highest diarrhea morbidity was observed in DJM75 (p<0.05). Phorbol esters were not detected in pig muscle tissues. Conclusion: The DJM was a good protein source for pigs, and could be used to replace SBM up to 30% (diet phorbol esters concentration at 5.5 mg/kg) in growing pig diets with no detrimental impacts on growth performance, nutrient utilization, and meat edibility.

식물체에 감염성 질병을 유발하는 바이로이드 검출 및 진단 방법 (The Detection and Diagnosis Methods of Infectious Viroids caused Plant Diseases)

  • 이세희;김양훈;안지영
    • 생명과학회지
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    • 제26권5호
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    • pp.620-631
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    • 2016
  • 바이로이드는 매우 작은 RNA 분자로 구성되어 있으며, 외피 단백질이 없고 오로지 식물에만 감염되어 질병을 유발한다. 바이로이드 감염 질병을 예방하거나 진단하는 것은 상당히 어려운 일이며, 이는 병징이 초기에는 발견되지 않고 수확기에 접어들어서 발견되기 때문이다. 한편, 혈청학적인 방법은 식물 병원체를 검출하기 위해 주로 사용되었으나 바이로이드는 핵산인 RNA로만 구성되어 있기 때문에 이 방법으로 검출할 수가 없다. 때문에 바이로이드를 검출하기 위해 주로 사용되는 방법은 분자 생물학적인 방법으로, 초기에는 바이로이드의 분자적인 크기와 구조적 특징을 이용한 겔 전기 영동 방법이 주로 사용되었다. 그 후에는 역전사 반응과 중합효소 연쇄반응을 접목시킨 역전사 중합효소 연쇄반응(RT-PCR) 방법이 활용되었고, 그에 대한 효율적인 결과 확인을 위해 형광 물질을 도입한 실시간 역전사 중합효소 연쇄반응(Real-time RT-PCR)이 도입되었다. 그러나 그들은 온도를 변화시키기 위한 값비싼 기기와 전문적인 인력이 필요함으로 현장에서는 활용되기가 어렵다. 최근 개발된 고리 기반의 등온 증폭법(Loop-mediated isothermal amplification)의 경우, 온도의 변화가 필요 없어 비싼 온도 조절 기기가 필요하지 않다. 또한 매우 높은 증폭 효율을 지니며 반응 시간이 짧은 등의 여러 장점을 지니고 있기에 최근 현장 진단용 기술에 도입되고 있다. 이러한 배경으로, 이 총설에서는 바이로이드 유발 질병에 대하여 요약하고 그에 대한 검출 및 진단 방법에 대한 연구 동향에 대하여 기술하였다.

닭 텔로미어 길이의 유전력 추정과 유전 전이 양상 (Inheritance and Heritability of Telomere Length in Chicken)

  • 박단비;손시환
    • 한국가금학회지
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    • 제41권3호
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    • pp.217-225
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    • 2014
  • 텔로미어는 진핵 세포의 염색체 말단으로, 직렬 반복 DNA 염기 서열과 shelterin 단백질 복합체로 구성되어 있다. 텔로미어의 기능은 염색체를 보호하는 것으로 체세포의 텔로미어 길이는 세포 분열시 DNA 복제 결실로 인해 연령이 증가함에 따라 감소하는 경향이 있다. 그러나 유전적, 후생유전학적 및 환경적 수준에서 여러 가지 요인이 텔로미어 길이에 영향을 미친다. 따라서 본 연구에서는 닭의 텔로미어 길이의 유전력을 추정하고, 이들의 유전전이 양상을 살펴보고자 하였다. 텔로미어 길이는 백혈구를 이용하여 양적 형광접합보인법(Q-FISH)과 양적 중합효소 연쇄반응법(qRT-PCR)으로 분석하였다. 분석 결과, 텔로미어 길이의 유전력은 자손과 부모 회귀 분석에 의해 출생 시 0.9로 추정되었고, 10 주령 및 30주령 때 부 분산 분석에 의해 0.03과 0.04로 추정되었다. 부와 자손 간(r=0.348) 및 모와 자손 간(r=0.380) 텔로미어 길이는 모두 유의한 정의 상관 관계를 보였다. 따라서 닭 텔로미어의 유전 전이 양상은 부모 양쪽 모두로부터 비슷하게 자식에 영향을 미치는 것으로 판단된다. 더불어 암수 자손에 미치는 영향 또한 유사한 것으로 나타났다. 이러한 결과는 부모의 텔로미어 길이의 각인이 성염색체의 유전자가 아닌 상염색체의 유전자에 의해 조절되는 것을 의미한다. 또한, 산모 연령에 따른 출생 자손의 텔로미어 길이는 차이가 없는 것으로 나타났다. 따라서 모체의 연령이 출생 자손의 텔로미어 길이에 영향을 미치지 않는데, 이는 수정란의 초기 배아 단계에서 세포적 reprogramming이 이루어지기 때문으로 사료된다.

산란성계에서 사료 급이량 조절이 생산성과 계육품질에 미치는 영향 (Effects of Regulate in Feed Intakes on Performance and Meat Quality in Old Laying Hens)

  • 강환구;김찬호
    • 한국가금학회지
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    • 제42권3호
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    • pp.205-214
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    • 2015
  • 본 연구는 100주령 이상의 산란성계에서 사료 급이량 조절에 따른 생산성, 도체 수율 및 육질을 주령에 따라 평가하고자 실시하였다. 공시 동물은 100주령 Hy-Line Brown 200수를 선별하여 A형 2단 케이지에 대조구 포함 총 5처리구로 구성하여 처리당 4반복, 반복당 10수씩(2수 수용 케이지 10개) 난괴법으로 임의 배치하였다. 시험 기간 동안 물은 자유 섭취하게 하였으며, 일반적인 점등관리(자연일조 + 조명; 16hr)를 실시하였다. 시험구는 대조구(일일 사료 섭취량 100%), 90%, 60%, 50%, 20% 총 5처리였다. 산란율은 사양 시험 기간 동안 일일 사료 섭취량이 감소함에 따라 산란율 및 사료 요구량은 유의적으로(P<0.05) 감소하였다. 도체 수율 역시 사양시험 기간 동안 일일 사료 섭취량이 감소함에 따라 도체 수율 및 부분육(가슴, 다리, 날개) 비율이 유의적으로(P<0.05) 감소하였다. Leukocyte 함량은 일일 사료 섭취량이 50%, 20%일 때 WBC, HE, LY, MO 및 EO 함량의 비율이 유의적으로(P<0.05) 높았다. 가슴육 내 일반 성분항목, 보수력, 가열감량 및 지방산 함량에 있어서는 사양시험 기간과 일일 사료 섭취량에 따른 유의적인 차이는 나타나지 않았다. 본 실험 결과 일일 사료 섭취량이 감소할수록 생산성과 도체율이 감소하였으나, 일일사료 섭취량을 60% 수준까지 낮춰도 무방하다고 생각되며, 계육 품질에 미치는 영향은 나타나지 않았다.