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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Enhancement of Tumor Response by MEK Inhibitor in Murine HCa-I Tumors (C3H/HeJ 마우스 간암에서 MEK 억제제에 의한 방사선 감수성 향상 효과)

  • Kim, Sung-Hee;Seong, Jin-Sil
    • Radiation Oncology Journal
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    • v.21 no.3
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    • pp.207-215
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    • 2003
  • Purpose: Extracellular signal-regulated kinase (ERK), which is part of the mitogen-activated protin kinase cascade, opposes initiation of the apoptotic cell death which is programmed by diverse cytotoxic stimuli. In this regard, the inhibition of ERK may be useful in improving the therapeutic efficacy of established anticancer agents. Materials and Methods: Murine hepatocarcinoma, HCa-I is known to be highly radioresistant with a TCD50 (radiation dose yield in $50\%$ cure) of more than 80 Gy. Various anticancer drugs have been found to enhance the radioresponse of this particular tumor but none were successful. The objective of this study was to explore whether the selective inhibition of MEK could potentiate the antitumor efficacy of radiation in vivo, particularly in the case on radioresistant tumor. C3H/HeJ mice hearing $7.5\~8\;mm$ HCa-I, were treated with PD98059(intratumoral injection of $0.16\;\mug/50\;\mul$). Results: Downregulation on ERK by PD98059 was most prominent 1h after the treatment. In the tumor growth delay assay, the drug was found to Increase the effect of the tumor radioresponse with an enhancement factor (EF) of 1.6 and 1.87. Combined treatment of 25 Gy radiation with PD98059 significantly increased radiation induced apoptosis. The peak apoptotic index (number on apoptotic nuclei in 1000 nuclei X100) was $1.2\%$ in the case of radiation treatment alone, $0.9\%$ in the case of drug treatment alone and $4.9\%,\;5.3\%$ in the combination treatment group. An analysis of apoptosis regulating molecules with Western blotting showed upregulation of p53, p$p21^{WAF1/CIP1}\;and\;Bcl-X_s$ in the combination treatment group as compared to their levels in either the radiation alone or drug alone treatment groups. The level of other molecules such as $Bcl-X_L4, Bax and Bcl-2 were changed to a lesser extent. Conclusion: The selective inhibition of MEK in combination with radiation therapy may have potential benefit in cancer treatment.

Effects of Depth and Duration of Flooding on Growth and Yield at Flowering Stage in Tomato(Lycopersicon esculentum). (토마토(Lycopersicon esculentum)의 개화기 침수 처리에 따른 생육 반응)

  • Guh, Ja-Ock;Han, Sung-Uk;Kuk, Yong-In;Chon, Sang-Uk;Lee, Young-Man
    • Korean Journal of Environmental Agriculture
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    • v.16 no.2
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    • pp.130-135
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    • 1997
  • Tomatoes are flooded differently 0, 5, 10 and 15cm, according to the developing stages such as flowering stage under the condition of greenhouse. Along with this, they are treated according to the time condition such as 6, 12, 24, 48 and 120 hours. The results obtained are summarized as follows. Plant height decreased in the depth of $0{\sim}10cm$ for over 48 hours, in the depth of 15cm for over 24 hours. Number of leaves was the same as in control, and it decreased over. Number of flowers and fruit setting of individuals decreased conspicuously according as the depth and the hours got greater and longer. Adventitious root occurred remarkably in the depth of $0{\sim}10cm$, for over 24 hours and in the depth of 15cm, 12 hours. Epinastic curvature increased greatly as the depth and the hours got greater and longer. Diffusion resistance of stomata cell increased as the depth and the hours got greater and longer. Diseases occurred conspicuously as the hours of flooding got longer rather than as the depth greater. The preventing of diseases caused by insecticide was observed, but it was not greater than in the seedling and transplanting stage. Fertilization was effective in the case of increasing the weight of shoot. Number of fruits per plant did not decrease in the depth of 0cm up to 24 hours, but decreased on the deeper level of flooding and increased as the hours got longer. Moreover with the exception of 120 hours per respective depth of the treatment, average weight of a fruit got greater as the depth and the hours got greater and longer. In the case of epinastic curvature and diffusion resistance, there was negative correlation between all the other investigated characters and positive correlation between weight of a fruits and average weight of a fruit.

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Studies on Dry Matter Yield s, Chmical Composition And Net Energy Accumlation in Three Leading Temperate Grass Species III. Seasonal changes of chemical components under dfferent cutting managements (주요북방형 목초의 건물수량 , 화학성분 및 Net Energy 축적에 관한 연구 III. 예취관리에 화학성분의 계절적 변화)

  • 김정갑;양종성;한흥전
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.6 no.3
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    • pp.157-163
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    • 1986
  • Synthesis and accumulation pattern of Weender components in orchardgrass (Dactylis glomerata L.) cv. Potomac and Baraula, perennial ryegrass (Lolium perenne L.) cv. Reveille and Semperweide and meadow fescue (Festuca pratensis Huds.) cv. Cosmos 11 and N.F.G. were studied under different growth environments and cutting managements. The field experiments were conducted as a split plot design with three cutting regimes of 6-7 cuts at grazing stage, 4-5 cuts at silage and 3 cuts at hay stage from 1975 to 1979 in Korea and West Germany. The results obtained are summarized as follows: 1. Air temperature, rainfalls and solar radiation were found to be an important meteorological factors influenced to synthesis and accumulation of Weender components. Under high temperature and strong solar radiation during summer season in Korea, accumulation of crude fiber and cell-wall constituents (NDF) in the plants, as average of all grass species and cutting regimes, were increased to about 30.1% and 48.7% from 27,9% and 42.9% in spring, respectively, while total nonstructural carbohydrates (TNC) were decreased to 1.52% in summer from 4.01% in spring. In West Germany, the concentration of Weeder components showed little seasonal variation. 2. Crude fiber and neutral detergent fiber (NDF) were shown higher concentration in orchardgrass than those of perennial ryegrass and meadow fescue, but N-free extractions and TNC as well as net energy value were less accumulated in orchardgrass. Orchardgrass contained lower net energy contents with 534 StE. 431 StE and 575 StE/kg for Suweon, Cheju and Freising, respectively, as compared with 624 StE (Suweon), 491 StE (Cheju) and 657 StE/kg (Freising) in meadow fescue.

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Characterization of Bacteriocin Produced from Isolated Strain of Bacillus sp. (Bacillus 속 분리주가 생산하는 박테리오신의 특성 조사)

  • Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.202-210
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    • 2017
  • As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

A Lodging Tolerant, Opaque Rice Cultivar 'Seolgaeng' (벼 내도복 뽀얀멥쌀 신품종 '설갱')

  • Hong, Ha-Cheol;Moon, Huhn-Pal;Choi, Hae-Chune;Hwang, Hung-Goo;Kim, Yeon-Gyu;Kim, Hong-Yeol;Yea, Jong-Doo;Shin, Young-Seop;Kang, Kyung-Ho;Choi, Yong-Hwan;Cho, Young-Chan;Baek, Man-Kee;Yang, Chang-Ihn;Choi, Im-Soo;Ahn, Sang-Nag;Yang, Sae-June
    • Korean Journal of Breeding Science
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    • v.43 no.6
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    • pp.532-537
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    • 2011
  • 'Seolgaeng' is a new Japonica rice variety developed from an 'Ilpumbyeo' mutant line, N-methyl-N-nitrosourea (MNU) treatment on fertilized egg cell, by a rice breeding team of National Crop Experiment Station, Rural Development Administration in 2001. This variety has about 134 days of growth duration from transplanting to harvesting in central plain area of Korea, a good semi-elect plant type and resistant to lodging with strong culm. The number of panicles/hill of 'Seolgaeng' is more than that of 'Hwaseongbyeo'. The milled rice appearance of this cultivar is an opaque, but it has amylose content (19.3%) similar to 'Hwaseongbyeo'. It is superior to hypae formation of Aspergillus orzyae in the making of fermented rice and amount of pigmentation in fermented rice by Monascus anka that of 'Ilpumbyeo'. This variety showed slow leaf senescence and considerable resistance to viviparous germination. It is moderately resistant to leaf blast and susceptible to bacterial blight, virus disease and insect pests. The yield performance of this rice cultivar is about 5.27 MT/ha in milled rice in local adaptability test for three years from 1999 to 2001). 'Seolgaeng' is adaptable to central and southern plain areas of Korea.

Potassium Physiology of Upland Crops (밭 작물(作物)의 가리(加里) 생리(生理))

  • Park, Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.10 no.3
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    • pp.103-134
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    • 1977
  • The physiological and biochemical role of potassium for upland crops according to recent research reports and the nutritional status of potassium in Korea were reviewed. Since physical and chemical characteristics of potassium ion are different from those of sodium, potassium can not completely be replaced by sodium and replacement must be limited to minimum possible functional area. Specific roles of potassium seem to keep fine structure of biological membranes such as thylacoid membrane of chloroplast in the most efficient form and to be allosteric effector and conformation controller of various enzymes principally in carbohydrate and protein metabolism. Potassium is essential to improve the efficiency of phoro- and oxidative- phosphorylation and involve deeply in all energy required metabolisms especially synthesis of organic matter and their translocation. Potassium has many important, physiological functions such as maintenance of osmotic pressure and optimum hydration of cell colloids, consequently uptake and translocation of water resulting in higher water use efficiency and of better subcellular environment for various physiological and biochemical activities. Potassium affects uptake and translocation of mineral nutrients and quality of products. potassium itself in products may become a quality criteria due to potassium essentiality for human beings. Potassium uptake is greatly decreased by low temperature and controlled by unknown feed back mechanism of potassium in plants. Thus the luxury absorption should be reconsidered. Total potassium content of upland soil in Korea is about 3% but the exchangeable one is about 0.3 me/100g soil. All upland crops require much potassium probably due to freezing and cold weather and also due to wet damage and drought caused by uneven rainfall pattern. In barley, potassium should be high at just before freezing and just after thawing and move into grain from heading for higher yield. Use efficiency of potassium was 27% for barley and 58% in old uplands, 46% in newly opened hilly lands for soybean. Soybean plant showed potassium deficiency symptom in various fields especially in newly opened hilly lands. Potassium criteria for normal growth appear 2% $K_2O$ and 1.0 K/(Ca+Mg) (content ratio) at flower bud initiation stage for soybean. Potassium requirement in plant was high in carrot, egg plant, chinese cabbage, red pepper, raddish and tomato. Potassium content in leaves was significantly correlated with yield in chinese cabbage. Sweet potato. greatly absorbed potassium subsequently affected potassium nutrition of the following crop. In the case of potassium deficiency, root showed the greatest difference in potassium content from that of normal indicating that deficiency damages root first. Potatoes and corn showed much higher potassium content in comparison with calcium and magnesium. Forage crops from ranges showed relatively high potassium content which was significantly and positively correlated with nitrogen, phosphorus and calcium content. Percentage of orchards (apple, pear, peach, grape, and orange) insufficient in potassium ranged from 16 to 25. The leaves and soils from the good apple and pear orchards showed higher potassium content than those from the poor ones. Critical ratio of $K_2O/(CaO+MgO)$ in mulberry leaves to escape from winter death of branch tip was 0.95. In the multiple croping system, exchangeable potassium in soils after one crop was affected by the previous crops and potassium uptake seemed to be related with soil organic matter providing soil moisture and aeration. Thus, the long term and quantitative investigation of various forms of potassium including total one are needed in relation to soil, weather and croping system. Potassium uptake and efficiency may be increased by topdressing, deep placement, slow-releasing or granular fertilizer application with the consideration of rainfall pattern. In all researches for nutritional explanation including potassium of crop yield reasonable and practicable nutritional indices will most easily be obtained through multifactor analysis.

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Studies on the Effects of Caponization and Various Hormone Treatment on the Meat Production and Quality in Growing Chicken (닭에 있어서 거세(去勢) 및 Hormone 처리(處理)가 산육성(産肉性) 및 육질(肉質)에 미치는 영향(影響)에 관한 연구(硏究))

  • Ra, Kwang Yon
    • Korean Journal of Agricultural Science
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    • v.2 no.1
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    • pp.9-47
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    • 1975
  • These experiments were caried out to study the effects of caponization and various hormone treatments upon meat production and improvement of meat quality of growing chicken. Sixtyseven days old 160 New Hampshire cockerels were treated and growth rate, carcass yield, change of weight of individual organs, meat composition and change of amino acid were measured and analysed. Otherwise change of testis and thyroid gland by hormone treatment were investigated histologically. The results obtained were as follows. 1. The effectst of caponization and hormone treatment upon meat production were; 1) Body weight of cockerels in D. E. S. group without caponization was increased. upon 96.86% than initial period and A. C. T. H. group was 104.22% but other groups and all carponization groups were lighter than those of control group. 2) Weekly body gain of D. E. S. group without caponization was best showing the significance (102.69 g) and the group with caponization were lower than those groups without caponization. 3) Carcass yield was best in Testo. group without caponization (831.2 g) and the group with caponization were lower than the group without caponization. 4) Carcass rate was highest in A. C. T. H. group with caponization and (67.22%) lowest in Testo. group without caponization (63.37%), but any significance was not recognized. 2. The effects of caponizatitn and hormone treatments upon the coposition of meat and amino acids were; 1) Any significance was not recognized between treated and untreated group about change of moisture, crude protein, crude ash and glycogen contents in meat. 2) Fat co tent in muscle in the all treated groups were higher than that of control group. 3) Extracts of group without caponization were higher than those of groups with caponization. 4) Lysin contents were highest in D. E. S. group with caponization (11. 12/ 16.0 g N) and generelly Testo. group was lower compared with D. E. S. group. 5) Histidine and Arginine contents were higher in the groups with caponization than without caponization. 6) Aspartic acid content were higher in D. E. S. group and A. C. T. H. group without depend on caponization. 7) Treonine content was higher in Testo. group without caponization and in the group with caponization and without hormone treatment compared with those of control group without caponization. 8) Serine content was decreased in the group with caponization and increased by D. E. S. and A. C. T. H treatment groups and glutamic acid was also decreased in Testo. group with out caponization. 9) Cystine content was decreased by Testo. treatment and was not appeared in Testo. group without caponization. 10) Valine content was lower in control group with caponization but significance was not recognized between other groups and control group without caponization. 11) Glycine, Alanine, Methionine. Isoleucine, Leucine, Thyrosine and Phenylalanine contents were not so difference between hormone treated groups and control group without caponization. 3. The effects of caponization and hormone treatment upon the change of organs were: 1) The weight of all organs were heaviest in D. E. S. group without caponization (18.5g) and lightest in A. C. T. H. group without caponization (155. 3g) but no significance was recognized between hormone treatment groups. 2) Heart weight was heaviest in D. E. S. group without caponization (7.46 g) and lightest in Testo. group without caponization (5.95 g). 3) Liver weight was heaviest in D. E. S. group without caponization(32.89g) and lightest in hormone untreated group with caponization(29.66g). Significance was not recognized. 4) Spleen weight was heaivest in Testo. group with caponization (3.22 g) and lightest in D. E. S. group without caponization(2.00g) in contrast with the other groups. High significance was recognized among the groups (P<0.01). 5) Cloacal thymus weight was lightest in D. E. S. group with or without caponization compared with control group without caponization. High significance was recognized among the groups. 6) Muscle fat content was not appeared in A. C. T. H. group with caponization, but it was highly increased in D. E. S. group with or without caponization. 7) Testis weight was lightest in D. E. S. group (0.38g) compared with control group (2.66g). Significance was recognized among the groups. 8) Large intestine, small intestine and cecum weight and length were heavier and longer in D. E. S. group without caponization and control group without caponization was lighter than those of hormone treated groups. 4. The effects of caponization and hormone treatment upon histological change of testis and thyroid gland: 1) The histological change of testis was significantly appeared in D. E. S. group that seminifirous tubles was slowly atrophied, the funtion of spernatogenesis was ceased, spermatocyte was changed as degeneration by pyknosis and karyorrhexis and interstitial cell was also atrophied, but in Testo. and A. C. T. H. group were similar as control group. 2) The histological change of thyroid gland in Testo. and A. C. T. H. groups without caponization were similar to that of control group without caponization, but in D. E. S. group without caponization, was changed squamously. Thyroid gland of the groups with caponization, epithelium of was atrophied and changed squamously as degeneration by pyknosis and karyorrhexis and the function of thyroid gland was slowly ceased in colloid and in hormone treated group with caponization.

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