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Studies on In Vitro Fertilization and Development of In Vitro Matured Porcine Follicular Oocytes I. Effect of Media and Capacitation Procedure on In Vitro Fertilization (체외성숙 돼지난포란의 체외수정과 배발달에 관한 연구 I. 배양액, 수정능획득 방법이 체외성숙 난포란의 체외성숙에 미치는 영향)

  • 정형민;엄상준;승경록;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.103-111
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    • 1993
  • These experiments were undertaken to establish the optimal culture systems for in vitro maturation, fertilization and subsequently embryonic development of porcine immature follicular oocytes isolated from the ovary of slaughtered pigs. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles (3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated (39$^{\circ}C$, 5% CO2 in air) for 42hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were prepared forfertilizaing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${mu}ell$ of capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the culture media. The fertilization rates of in vitro matured follicular oocytes cultured in B. O., TCM-HEPES, m-KRB, and TALP-II media were 61.3%, 83.0%, 88.9% and 89.2%, respectively. In addition, the polyspermy rates were 60.7%, 66.5%, 53.8%, and 43.9%, respectively. These data indicated that the highest of fertilization and the lowest of polyspermy rate was shown in TALP-II medium. Spermatozoa capacitated by caffeine, heparin, and percoll density gradient treatment in the 4 different media, the fertilization rates were 33.0~57.2%, 39.9~90.2%, and 52.6~92.8%, respectively, showing the lowest rate in caffeine treatment. The development rate of follicular oocytes, fertilized with the spermatozoa capacitated by caffeine, heparin, and percoll gradient in the TALP-II medium, upto 2 to 4-cell stages were 32.6%, 74.5% and 70.9%, respectively. Finally, fertilization rates of follicular oocytes cultured with follicular fluid containing medium from 10 to 100% were 61.2~94.1% and the rates (90~94%) with 10~20% follicular fluids were significantly higher than those (85.3%) of cultured in the media without follicular fluid. In addition, the rates of pronucleus formation were also higher in follicular fluid treated group (73.1~83.0%) than those (64.7%) of oocytes cultured without follicular fluid. The highest fertilization and pronucleus formation rates was found in oocytes cultured with 10% follicular fluid. These results suggest that the addition of heparin or percoll density gradient method is better capacitation method. Furthermore, the addition of porcine follicular fluid to the fertilization medium may improve the fertilization rates and formation of pronucleus.

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Comparison of Developmental Competency of Porcine Embryos Cloned with Mesenchymal Stem Cells and Somatic Cells

  • Jin Hai-Feng;Kumar B. Mohana;Cho Sung-Keun;Ock Sun-A;Jeon Byeong-Gyun;Balasubramanian S.;Choe Sang-Yong;Rho Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.30 no.2
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    • pp.119-124
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    • 2006
  • The present study compared the developmental potential of cloned porcine embryos with mesenchymal stem cells (MSCs), fetal fibroblasts (FFs) and cumulus cells (CCs) by assessing the cleavage and blastocyst rate, total cell number, inner cell mass (ICM) ratio and apoptosis. MSCs were isolated by ficoll gradients from femur of -6 month old female pig, and maintained for primary cultures. FFs from a female fetus at ${\sim}30$ day of gestation were established, and CCs were obtained from cumulus oocyte complexes (COCs) aspirated from $3{\sim}6$ mm follicles in diameter. Donor cells at $3{\sim}4$ passage were employed for nuclear transfer (NT). COCs were matured and fertilized in vitro(IVF) as control. Cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs, FFs and CCs ($82.7{\pm}8.9%\;vs\;70.6{\pm}5.4,\;68.7{\pm}5.1\;and\;63.4{\pm}5.6%$, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs ($24.5{\pm}2.8\;and\;20.4{\pm}8.3%$) did not differ, but were significantly (P<0.05) higher than NT derived from FFs and CCs ($10.6{\pm}2.7\;and\;9.8{\pm}2.1%$). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs ($35.4{\pm}5.2\;and\;0.40{\pm}0.09%$, respectively) were significantly (P<0.05) higher than those from FFs and CCs ($24.9{\pm}6.2%\;vs\;0.19{\pm}0.16,\;23.6{\pm}5.5\;and\;0.17{\pm}0.16%$, respectively). Proportions of TUNEL positive cells in NT embryos from FFs and CCs ($6.9{\pm}1.5\;and\;7.4{\pm}1.7%$, respectively) were significantly (P<0.05) higher than in MSCs ($4.8{\pm}1.4%$) and IVF ($2.3{\pm}0.9%$). The results demonstrate that MSCs have a greater potential as donor cells than FFs and CCs in achieving enhanced production of cloned porcine embryos.

Response of Monodehydroascorbate Reductase (MDHAR) in Lettuce (Lactuca sativa L.) Leaves Subjected to Water Deficit Stress (수분 부족 스트레스 처리시 Monodehydroascorbate Reductase (MDHAR)의 반응)

  • Kang, Sang-Jae
    • Journal of Bio-Environment Control
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    • v.17 no.4
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    • pp.273-282
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    • 2008
  • The relationship between water deficit stress and monodehydroascorbate reductase (MDHAR) activity was determined in lettuce (Lactuca sativa L.) leaves under water stress condition imposed by with-holding water for 72 hrs. Relative water content determined in water deficit stressed lettuce leaves gradually reduced from 91.29% to 74.58%, and water content of medium drastically decreased 4.73% after quitting of irrigation. Hydrogen peroxide content in leaves subjected to water deficit stress rapidly increased, but soluble protein content rapidly decreased when those were compared to control plant. The relationship between relative water content and hydrogen peroxide content in stressed leaves positively correlated with $R^2$=0.8851, but soluble protein content reversely correlated with $R^2$=0.9826. Total chlorophyll content in stressed plant leaves was higher than that of control plant, and increased rapidly in early stage of treatment of both stressed and control plants. Carotenoid content was higher than that of control plant, and the ratio of carotenoid to total chlorophyll in stressed plant was higher as compared to control plant. As water deficit stress continued progressively, total ascorbate content in stressed plant leaves was a little higher than that of control plant. But dehydroascorbate (DHA) content within 6 hr of water deficit stress was higher than that of control plant, and then, content of control plant in 12 hr of stress treatment higher than that of stressed leaves. The activity of monodehydroascorbate reductase of cytosolic and chloroplastic tractions increased dramatically, and mRNA of MDHAR was highly detected by probing $^{32}P$-labeled single stranded MDHAR RNA of lettuce plant leaves subjected to water deficit stress. Relationship between MDHAR activity and relative water content and hydrogen peroxide highly correlated with $R^2$=0.9937 and 0.8645, respectively.

The Effect of Dietary n-3 and n-6 Polyunsaturated Fats on changes in Glucose, Non Esterified Fatty Acid and Fatty Acid Compositions in Serum of Rat Exposed to Stress. (N-3계 및 N-6계 지방산 식이가 스트레스에 노출된 흰 쥐의 혈당과 혈청 유리지방산 및 지방산 조성변화에 미치는 영향)

  • 장문정
    • Journal of Nutrition and Health
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    • v.28 no.5
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    • pp.375-386
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    • 1995
  • This study was designed to investigate the changes in energy substrates, glucose and non-esterified fatty acid(NEFA), and fatty acid compositions in serum, following physiolgical stress in rats fed diets containing various fatty acids. Forty two Sprague-Dawley strain male rats, weighing 108$\pm$2.1g, were fed 3 different experimental diets for 4 weeks. The diets were composed of 105 fat(w/w) of either corn oil(CO;18:2 n6:57%), plant perilla oil(PO;18:3 n3:59%), or tuna fish oil(FO;20:5 n3:17%%, 22:6 n3:19%). After 4 weeks of feeding, each group wa subdiveided into (a) control, (b) 2 min swim in ice-cold water. Animals wer decapitated 20min after commencing the swim; trunk blood, brain, liver and epididymal fat pad were obtained. The levels of serum corticosterone, glucose, NEFA, triglyceride, fatty acid compositions, brain serotonin and 5-hydroxyindoleacetic acid were determined. Basal levels of corticosterone na NEFA of serum were significantly lower in fish oil fed animals than those of any other oil fed animals. Compared to either perilla oil-fed or corn oil-fed rats, cold swim stress in fish oil fed rats produced significantly smaller NEFA and larger corticosterone responses. However, there was no significant difference in basal levels of serum glucose. Stress increased serum glucose levels slightly, and the amount of increment was larger in fish oil rats than those of any other oil fed rats than those of any other oil fed rats, although all the values were normal level. Dietary fats and stress did not affect serotonin metabolism. In additions, the composition of fatty acids in serum was significantly affected by the dietary compostion of fatty acids and stress. Stress induced decreases in monounsaturated fatty acid and non-polyunsaturated fatty acid concentration in either perilla oil fed or fish group, but did not in corn oil fed group. Stress resulted in changes in fatty acid metabolism similar to that associated with essential fatty acid(EFA) dificiency, when feeding animals n-3 fatty acids in diet. In conclusion, feeding fish oil was more effective to decrease NEFA in serum than feeding perilla oil or corn oil and improved lipid metabolism, when the rats were maintained in normal or exposed to stressful environment. However, the fact that feeding diet containing n-3 fatty acids decreased EFA status under stress suggests that the requirement of n-6 PUFA should be increased in these groups.

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STUDIES ON THE NUTRIENT REQUIREMENTS OF KOREAN NATIVE CATTLE I. EFFECT OF ENERGY LEVELS ON PERFORMANCE AND ENERGY INTAKE OF GROWING AND FINISHING KOREAN NATIVE CATTLE

  • Ahn, B.H.;Ahn, D.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.2 no.2
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    • pp.59-66
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    • 1989
  • Growing and finishing Korean native bulls were used to investigate the effects of different dietary energy levels on performance and energy intake. Experimental observations were made over three weight categories (200 to 250 kg, 250 to 350 kg and 350 to 450 kg). Three diets (2.4, 2.6 and 2.8 Mcal ME/kg DM) were used for each weight category. Crude protein levels of the diets were 12, 11 and 10% for the respective weight groups. Dietary energy level did not significantly affect daily body gain within a weight range but daily body gain during the entire experiment was higher (P<.05) in bulls receiving 2.6 Mcal energy diet than in those fed 2.4 and 2.8 Mcal energy diets. The following equation was developed to predict daily body gain(Y) from energy levels(X) of ration. $Y\;=\;1.3.475X\;-\;2.5949X^{2}\;-\;16.355$ Increasing energy levels significantly (P<.05) decreased daily feed intake. The following equation was developed to predict daily feed intake(Y) from energy levels(X) of ration. $Y\;=\;-30.013X\;+\;5.4401X^{2}\;+\;49.119$ Feed intake per metabolic body size during the entire feeding period ranged from 100.9 to 110.8 g and was lower in bulls fed 2.6 and 2.8 Mcal energy diets than in those fed 2.4 Mcal energy diet. Increasing energy levels significantly (P<.05) improved feed efficiency. The following equation was developed to predict feed efficiency(Y) from energy levels(X) of ration. $Y\;=\;-118.34X\;+\;22.448X^{2}\;+\;162.85$ Daily energy intake during the entire experiment ranged from 18.90 to 19.99 Mcal and there was no significant difference among energy levels. Daily energy intake per metabolic body size during the feeding period ranged from 248.6 to 260.8 kcal and was slightly higher in bulls receiving 2.8 Mcal than in those fed 2.4 and 2.6 Mcal energy diets. Energy required per kg body gain ranged from 17.25 to 19.11 Mcal and was slightly lower in bulls receiving 2.6 Mcal energy diet than in those fed 2.4 and 2.8 Mcal energy diets.

Effect of different levels of protein concentrates supplementation on the growth performance, plasma amino acids profile and mTOR cascade genes expression in early-weaned yak calves

  • Peng, Q.H.;Khan, N.A.;Xue, B.;Yan, T.H.;Wang, Z.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.218-224
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    • 2018
  • Objective: This study evaluated the effects of different levels of protein concentrate supplementation on the growth performance of yak calves, and correlated the growth rate to changes occurring in the plasma- amino acids, -insulin profile, and signaling activity of mammalian target of rapamycin (mTOR) cascade to characterize the mechanism through which the protein synthesis can be improved in early weaned yaks. Methods: For this study, 48 early (3 months old) weaned yak calves were selected, and assigned into four dietary treatments according to randomized complete block design. The four blocks were balanced for body weight and sex. The yaks were either grazed on natural pasture (control diet) in a single herd or the grazing yaks was supplemented with one of the three protein rich supplements containing low (17%; LP), medium (19%; MP), or high (21%; HP) levels of crude proteins for a period of 30 days. Results: Results showed that the average daily gain of calves increased (0.14 vs 0.23-0.26 kg; p<0.05) with protein concentrates supplementation. The concentration of plasma methionine increased (p<0.05; 8.6 vs $10.1-12.4{\mu}mol/L$), while those of serine and tyrosine did not change (p>0.05) when the grazing calves were supplemented with protein concentrates. Compared to control diet, the insulin level of calves increased (p<0.05; 1.86 vs $2.16-2.54{\mu}IU/mL$) with supplementation of protein concentrates. Addition of protein concentrates up-regulated (p<0.05) expression of mTOR-raptor, mammalian vacuolar protein sorting 34 homolog, the translational regulators eukaryotic translation initiation factor 4E binding protein 1, and S6 kinase 1 genes in both Longissimus dorsi and semitendinosus. In contrast, the expression of sequestosome 1 was down-regulated in the concentrate supplemented calves. Conclusion: Our results show that protein supplementation improves the growth performance of early weaned yak calves, and that plasma methionine and insulin concentrations were the key mediator for gene expression and protein deposition in the muscles.

IM-133N - A Useful Herbal Combination for Eradicating Disease-triggering Pathogens in Mice via Immunotherapeutic Mechanisms

  • Firashathulla, Syed;Inamdar, Mohammed Naseeruddin;Rafiq, Mohamed;Viswanatha, Gollapalle Lakshminarayanashastry;Kumar, Lakkavalli Mohan Sharath;Babu, Uddagiri Venkanna;Ramakrishnan, Shyam;Paramesh, Rangesh
    • Journal of Pharmacopuncture
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    • v.19 no.1
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    • pp.21-27
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    • 2016
  • Objectives: The present study was undertaken to evaluate the immunomodulatory (IM) activity of IM-133N, a herbal combination in various immunotherapeutic experimental models. Methods: The IM activity of IM-133N was evaluated against three experimental models namely, effect of IM-133N against Escherichia coli (E. coli)-induced abdominal sepsis in mice, and carbon clearance test was performed in Wistar albino rats to evaluated the phagocytic potential of IM-133N, in addition IM-133N was evaluated for its immunoglobulin enhancing potential in rats, where the immunoglobulin levels were measured by zinc sulphate turbity (ZST) test. Further, IM-133N was subjected for detailed liquid chromatography-mass spectrometry (LC-MS)/MS analysis to identify the probable active constituents present in it. Results: The findings of the present study has demonstrated very promising IM property of IM-133N in all the experimental models. Briefly, pretreatment with IM-133N at 125, 250, 500 and 1,000 mg/kg, p.o. doses had protected the mice against E. coli-induced abdominal sepsis and mortality, further the effect of IM-133N was found to be significant and dose-dependent. In support of this, in another study administration of IM-133N showed a significant and dose-dependent increase in serum immunoglobulin levels, estimated by ZST test. In line with the above findings, in the carbon clearance test the low doses (125 and 250 mg/kg, p.o.) of IM-133N increased the rate of carbon clearance, whereas the higher doses (500 and 1,000 mg/kg, p.o.) did not sustain the response, and saturation effect was considered as one of the possible reason for futility of higher doses for IM-133N. In addition, A detailed LC-MS/MS analysis of IM-133N showed 17 bioactive phytochemical constituents: namely, apigenin, chaulmoogric acid, mesquitol, quercetin, symphoxanthone, salireposide, ${\beta}$-sitosterol, nonaeicosanol, ${\beta}$-amyrin, betulic acid, oleanolic acid, symplososide, symponoside, symploveroside, symplocomoside, symconoside A and locoracemoside B. Conclusion: These findings suggest that IM-133N possesses significant IM activity and, hence, could be useful for eradicating opportunistic disease-triggering pathogens via immunotherapeutic mechanisms. The findings also suggest IM-133N may also useful in other immunity disorders.

A study on Chinese Consumer Behavior of Korean Automobiles (중국소비자의 한국자동차 구매의도에 관한 연구)

  • SONG, Yun-Tao;LEE, Jong-Ho
    • The Journal of Industrial Distribution & Business
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    • v.10 no.6
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    • pp.29-37
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    • 2019
  • Purpose - This study intends to analyze Chinese consumers' purchase intention of Korean automobile. The aims of this study investigated to find out how product attributes and brand image affect the consumer's purchase intention of Korean automobiles in China. We consider the product attributes, brand image as evaluation criteria, also recognized as objectives for purchase intention. This study aims to analyze the positive effects on Chinese consumers' purchase intention with the variables by using the research model. Research design, data, and methodology - Based on theory of Purchase Intention, the concept of purchase intention was assessed by two factors: a) product attributes, and b) brand image to test the proposed hypotheses. The survey was conducted for 30 days, from October 17 to November 16 in 2018. With IBM SPSS Statistics 23.0 and IBM SPSS AMOS 23.0, factor analysis and regression, Cronbach's alpha and correlation were tested. 300 samples were used for this analysis. Results - As a main result, product attributes and brand image exerted significant effects on consumer's purchase intention. Thus,

    and

    were supported. The results showed that the product attributes, brand image had positive effects on purchase intention. The findings of this thesis are as follows: Firstly, automobiles' quality factors were divided into two categories like product properties (price, performance, design, economy, and safety), and brand image (awareness, favorability, reliability, and familiarity). Second, it showed that product properties and brand image had positive impacts on purchase intention of consumer. Conclusions - The impact of product attributes and brand image on consumer perceived value of Korean automobiles in China can be seen as a universal psychology of consumers who trust automobile products such as high quality, technology, ets. The results offer some insights into the extended model and have important managerial implications for Korean automobile enterprise. As the findings of this thesis show, all strategic orientations have significant and positive effects on the ability of innovation. At the same time, automobile company must pay attention to quality. As the relationship between the automobiles' performance and brand image were positive and significant, automobile companies should have comprehensive strategic orientations focused on strengthening these factors.

Quality Characteristics of Salt-Fermented Anchovy Sauce and Sandlance Sauce (멸치액젓 및 까나리액젓의 품질 특성)

  • OH Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.3
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    • pp.252-255
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    • 1999
  • The quality characteristics of the Korean traditional salt-fermented fish sauces, the traditional anchovy sauce (TAS) and the sandlance sauce(TKS) were evaluated comparing to the commercial anchovy sauce (CAS) and sandlance sauce (CKS). The acidity was higher in TKS than in TAS, whereas the contents of VBN, total-N and amino-N were higher levels in TAS. In color values, L and b values in TAS were generally higher than those in TKS, whereas a and ${\Delta}E$ values were higher in TKS. The contents of total free amino acids in TAS and TKS were $12.40\;g\%$ (w/v) and $9.549 g\%$ (w/v), respectively. The contents or six amino acids, alanine, glutamic acid, leucine, isoleucine, valine and Iysine were higher in TAS, whereas the contents of arginine, glutamic acid, leucine, alanine and valine were higher in TKS. Nucleotides such as IMP and hypoxanthine were principal components in both TAS and TKS. The nitrogen related compounds, TMAO, TMA and total creatinine were determined to be $108.8\;mg\%$ (w/v), $60.5\;mg\%$ (w/v), $62.4\;mg\%$ (w/v) in TAS, and those in TKS were $60.1\;mg\%$ (w/v), $24.1\;mg\%$ (w/v), $67.6\;mg\%$ (w/v), respectively.

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Community Structure and Distribution Pattern of the Pleuronectiform Fishes in the Uljin Marine Ranching Area, Korea (울진바다목장에서 어획된 가자미목(Pleuronectiformes) 어류의 군집구조 및 공간분포)

  • Yoon, Byoung Sun;Park, Jeong-Ho;Sohn, Myong Ho;Yang, Jae Hyeong;Yoon, Sang Chul;Choi, Young Min
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.4
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    • pp.413-423
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    • 2013
  • To elucidate the variation of species composition, biomass and length distribution of flatfishes by the period and region in the Uljin marine ranching area, bottom trawl survey was investigated at the six stations from Feb. 2008 to Dec. 2010. During the survey period, a total 4 families 17 species in the Pleuronectiformes, average 69,158 $ind./km^2$ and 5,625 $kg/km^2$ were identified, in 2008 appeared in 14 species, average 25,798 $ind./km^2$ and 2,333 $kg/km^2$ and in 2009 appeared in 13 species, average 102,360 $ind./km^2$ and 5,634 $kg/km^2$ and in 2010 appeared in 14 species, average 75,704 $ind./km^2$ and 8,632 $kg/km^2$. The individual dominant species, occupying over 10% of total individuals, was Pleuronectes herzensteini (20,811 ind., 30.0%), Hippoglossoides pinetorum (18,666 ind., 26.9%) and Glyptocephalus stelleri (13,499 ind., 19.4%) also the biomass dominant species, occuping over 10% of total biomass, was P. herzensteini (2,207 kg, 39.3%), Pleuronectes yokohamae (857 kg, 15.3%), H. pinetorum (761 kg, 13.5%), Kareius bicoloratus (677 kg, 12.1%). From the cluster and MDS analysis based on Bray-Curtis similarity matrix of fourth root transformed data of species number and individuals in the Uljin marine ranching area from Feb. 2008 to Dec. 2010 was divided into two different groups of the flatfishes community in 2008 and from Jan. to Apr. in 2009 and 2010 (Group A) and the pleuronectiform community in from May to Dec. in 2009 and 2010 (Group B). From the cluster and MDS analysis using the similarity of demersal organisms community among six stations, the Uljin marine ranching area was divided into two different groups of Group 1 (St. 1 and St. 3) and Group 2 (St. 2, St. 4, St. 5 and St. 6).