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Production of Monoclonal Antibodies Against the Immunoglobulin M of Olive Flounder Paralichthys Olivaceus (넙치(Paralichthys olivaceus)의 immunoglobulin M에 대한 단클론 항체 생산)

  • Kim, Wi-Sik;Kim, Ki-Hong;Kim, Choon-sup;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.2
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    • pp.169-174
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    • 2017
  • Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.

Optimization of Spinach Pesto by Response Surface Methodology (반응표면분석법을 이용한 시금치 페스토 제조조건의 최적화)

  • Kim, Ok-Sun;Park, Jong-Dae;Kum, Jun-Seok;Choi, Yun-Sang;Choi, Hyun-Wook;Sung, Jung-Min
    • The Korean Journal of Food And Nutrition
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    • v.29 no.4
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    • pp.583-594
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    • 2016
  • The purpose of this study was to determine the optimal mixing ratio of olive oil, parmesan cheese, and nuts for preparation of pesto with spinach. Based on a surface response methodology (RSM), the independent variables were olive oil (100~300 g), parmesan cheese (25~75 g) and nuts (12~36 g). The dependent variables were physico-chemical properties as pH, color values (L, a, and b values), viscosity and sensory evaluation. pH decreased with increasing parmesan cheese levels. The L value decreased and the a and b values increased with increasing olive oil levels. Viscosity was negatively correlated with olive oil levels but positively correlated with the amounts of parmesan cheese and nuts. The addition of oil exerted a positive effect on gloss, and oily taste. In the sensory evaluation, the values of appearance, color, flavor, taste and overall acceptance were in the range of 4.4~7.2, 3.7~7.4, 4.4~7.1, 3.9~7.3 and 3.5~7.6, respectively. The sensory evaluation results showed significant values in appearance (p<0.01), flavor (p<0.01), taste (p<0.01) and overall acceptance (p<0.05). The optimal amounts of spinach pesto with olive oil, parmesan cheese, and nuts were determined by numerical optimization of a canonical model and graphical optimization. The optimal amounts were 175.29 g of olive oil, 49.51 g parmesan cheese, and 27.37 g of nuts per 100 g of spinach.

Status of Marine Environment of Olive Flounder, Paralichythys olivaceus, Culture Ground in Jeju-do - Focus on Kudoa septempunctata positive and negative farm - (제주도 육상 넙치양식장의 해양환경현황 - 쿠도아 양성양식장과 음성양식장을 중심으로 -)

  • OH, Hyun-Taik;YI, Yong-Min;CHO, Yun-Sik;KIM, Jin-Ho;LEE, Kyu-Hyun
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.3
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    • pp.746-759
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    • 2016
  • In this study, we monitored the status of marine environment of olive flounder, Paralichythys olivaceus, culture ground in Jeju-do, Republic of Korea. It reported the consumption of raw olive flounder meat containing of Kudoa, Kudoa septempunctata, could induce vomitting and diarrhea in Japan. The Kudoa is a new mycosporean species, researchers found Kudoa from the muscles of olive flounder cultured at western Japan and imported from Jeju-do. We choose two Kudoa positive farms and two negative farms in the mid of concern about the relation between Kudoa and habitat. We found two marine invasive species at the water-pumping seabed at one of the Kudoa positive farm. The concentration of pH and DO at on-growing flounder farms showed the decreasing pattern along side the raceway (influent sea waters > on growing sea waters > outfluent sea waters). The TN and TP values increased gradually following to the raceway (influent sea waters < on growing sea waters < outfluent sea waters). The concentration of COD and SS were in the range of $0.100-2.581mg{\cdot}L^{-1}$, $1.00-12.70mg{\cdot}L^{-1}$, respectively. The calculated residence time was 4hr 32minutes at F1, 11hr 21minutes at F2, and 9hr 50minutes at F3, respectively. It was calculated same distance of 4 km away from effluent pipes. Although direct relation between Kudoa and marine environment could not define well based on this study result, the more studies on marine environmental stressors for olive flounder are required to conduct as a reliable method including socio-economic group and environmental group.

Effects of Dietary Supplementation of Cottonseed and Soybean meal on Reproductive Histology of Olive Flounder, Paralichthys olivaceus (사료 내 면실박 및 대두박 첨가가 넙치의 번식 조직학에 미치는 영향)

  • Lim Se-Jin;Lee Kyeong-Jun;Lee Young-Don;Song Young-Bo
    • Journal of Aquaculture
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    • v.19 no.2
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    • pp.133-139
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    • 2006
  • The gossypol existed in cottonseed meal is a well known antispermatogenic agent which can impair reproductive performances of male fish as well as mammals. Two feeding experiments were conducted to examine a toxic effect of dietary supplementation of cottonseed meal on reproduction in juvenile olive flounder (the first experiment) for 19 weeks and growing olive flounder (the second experiment) for 26 weeks. After each feeding study, females and males were sampled for histological examination in gonads and liver to verify any negative effects by the dietary supplementation of cottonseed and soybean meal on reproduction. After two feeding trial, the gonad somatic index (GSI) of male and female (from the first feeding trial) were not significantly different among all the dietary treatments. The GSI values of female (from the second feeding trial) were not significantly different among all the dietary treatments. However, males fed cottonseed and soybean meal containing diets exhibited significantly lower GSI than that fed the control diet after the second feeding trial. Histological examination of gonads and liver of fish fed cottonseed and soybean meal did not show any negative effects compared to those of fish ffd the control diet. Hepatosomatic index of fish in the first and second feeding trials were not significantly different among all the dietary treatments. The findings in this study suggest that dietary supplementation of cottonseed and soybean meal up to 40% fish meal replacement might not deteriorate the gametogenesis of juvenile and growing olive flounder. However, the supplementation in diets over 30% fish meal replacement might reduce GSI of male in growing olive flounder.

Processing and Quality Properties of Olive Flounder Paralichthys olivaceus Steak Added with Pork Leg (돼지후지육 첨가 넙치(Paralichthys olivaceus) 스테이크의 제조 및 품질 특성)

  • Yoon, Moon-Joo;Lee, Jae-Dong;Kang, Kyung-Hun;Park, Si-Young;Joo, Jong-Chan;Kim, Jeong-Gyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.6
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    • pp.849-856
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    • 2015
  • This study was performed to obtain basic data regarding the development of fish steak products using olive flounder and pork leg. Olive flounder and pork leg were ground separately in a chopper. The methods used for processing were as follows. Chopped olive flounder (100 g) and other ingredients (bread crumbs, 13 g; onion, 12 g; garlic, 4 g; egg wash, 18 g; salt, 0.05 g; pepper, 0.05 g) were mixed in a chopper. The mixture was molded into a steak shape ($12{\times}7cm$) and roasted in an oven at $180^{\circ}C$ for 12 min (OF). FP consisting of a mixture of olive flounder (70 g) and pork leg meat (30 g) and OP consisting of pork leg meat alone (100 g) were processed according to the same procedure as described for OF. Various factors (viable bacterial count, chemical composition, pH, salinity, hardness value, color value, total amino acid content, free amino acid content, fatty acid composition, mineral content) were measured, and sensory evaluation was conducted. Based on the results of the sensory evaluation and hardness value, OP was deemed to be the most desirable, followed in order by FP and OF. There was a slight but significant difference between OP and FP.

Evaluation of a Hydrolyzed Pig Bristle Meal as a Partial Fish Meal Replacer in Diets for Juvenile Olive Flounder Paralichthys olivaceus (치어기 넙치(Paralichthys olivaceus) 사료 내 돈모분(Pig Bristle Meal)의 어분대체 가능성 평가)

  • Kim, Youjeong;Shin, Jaehyeong;Kwon, Hwangwon;Lee, Soyoon;Kim, Joo-Min;Kim, Min-Gi;Kim, Jeong-Dae;Lee, Kyeong-Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.2
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    • pp.148-156
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    • 2018
  • This study was conducted to evaluate dietary hydrolyzed pig bristle meal (PBM) for juvenile olive flounder Paralichthys olivaceus. In Experiment 1 (EXP-1), six experimental diets were prepared to contain 0, 3, 6, 9, 12 and 15% PBM (designated Con, PBM3, PBM6, PBM9, PBM12 and PBM15, respectively). Triplicate groups of olive flounder (initial body weight, 8.69 g) were fed the diets to apparent satiation for 8 weeks during the optimal water temperature season ($20.5{\pm}2.12^{\circ}C$). All PBM supplemented groups except for PBM3 showed significantly lower growth performance and feed utilization compared to the control group. The protein digestibility of PBM3, PBM6, and PBM9 diets did not significantly differ from that of the control diet. In Experiment 2, 1% mono-calcium phosphate was added into the experimental diets used in Exp-1. Triplicate groups of olive flounder (10.6 g) were fed the diets to apparent satiation for 8 weeks during the low water temperature season ($12.5{\pm}1.12^{\circ}C$). The growth performances and feed utilization of fish fed all diets except for PBM15 diet did not significantly differ from those of the control diet. This study indicates that hydrolyzed PBM can replace fish meal by up to 12% with limiting amino acids and mono-calcium phosphate in diets for juvenile olive flounder.

Polymorphism Analysis of the ND-4 Gene for the Origin Determination of Olive Flounder, Paralichthys olivaceus. (넙치의 원산지 판별을 위한 ND-4유전자의 다양성 분석)

  • 송인선;진덕희;최석정;이석근
    • Journal of Life Science
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    • v.14 no.4
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    • pp.627-635
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    • 2004
  • In order to distinguish the genetic polymorphism among the olive flounder (Paralichthys olivaceus) obtained from East sea of Jumunjin, aquaculture of Tongyoung and Geoje, and East sea of North Korea, the ND-4 and cytochrome b genes of olive flounder were divided into 5 regions. Each region was analyzed by degenerating gel electrophoresis scanning (DGES), and by subsequent DNA sequencing. The DGES disclosed characteristic DNA polymorphisms in ND-4-2 and ND-4-3 regions of olive flounder, which were also confirmed by the DNA sequencing. The olive flounders obtained from the different marine areas showed DNA mutations in ND-4-2 region (G390A, C402T, and A411G; GenBank: AB028664), and also showed frequent DNA mutations in ND-4-3 region (C515G, C537T, C538T, A567G, G714A, C736T, G756A, A759T, T817C, and T829G), white the cytochrome b gene showed no DNA mutation both in the DGES and DNA sequencing. These data suggest that the ND-4-2 and ND-4-3 regions are candidate loci to distinguish the origin of olive flounder, and that the DGES used in this study provided fast and reliable informations for the genetic polymorphism.

Transcriptional analysis of olive flounder lectins in response to VHSV infection

  • Lee, Young Mee;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Noh, Gyeong Eon;Kim, Woo-Jin;Kim, Kyung-Kil
    • Journal of fish pathology
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    • v.29 no.1
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    • pp.13-23
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    • 2016
  • Lectins play significant roles in the innate immune responses through binding to pathogen-associated molecular patterns (PAMPs) on the surfaces of microorganisms. In the present study, tissue distribution and expression analysis of olive flounder lectins were performed after viral hemorrhagic septicemia virus (VHSV) challenge. Fish egg lectin and serum lectin were found to be predominantly expressed in the gills and liver, these results indicate that the transcript expression of olive flounder lectins is concentrated in immune-related tissues. Following a VHSV challenge, an overall increase in the transcript levels of the genes was observed and the expression patterns were distinctly divided into early and later responses during VHSV infection. In conclusion, olive flounder lectins are specifically expressed in immune-related organs and induced in both the immediate and long-lasting immune responses to VHSV in the olive flounder. These results indicate that lectins may be play important roles in the host defense mechanism and involved in the innate and adaptive immune response to viruses in fish.

Characterization of the Myxosporean Parasite Isolated from Emaciated Olive Flounders Paralichthys olivaceus on Jeju Island (제주도 여윔증상 넙치(Paralichthys olivaceus)로부터 분리한 점액포자충의 특성 분석)

  • Kim, Seung Min;Jun, Lyu Jin;Park, Myoung Ae;Jeong, Hyun Do;Jeong, Joon Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.3
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    • pp.337-345
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    • 2015
  • To investigate the causes of emaciation in cultured olive flounder Paralichthys olivaceus in Korea. We performed histological examinations and polymerase chain reaction (PCR) with a new primer set. In most cases, the most severe emaciation was observed in the abdominal area Using PCR on extracted livers, kidneys, spleens, gills, brains, and intestines, we found that areas around the kidneys and intestines were as almost always positive. In significantly emaciated fish, PCR was positive in all internal organs except the gills. In addition, the homology of 812-bp nucleotide sequences of the 28S rRNA gene was more than 99% in emaciated fish. Partial homology with Myxobolus spp. and Cystodiscus axonis, whose data were obtained from GenBank was 86% and 88%, respectively. Histological examinations detected spores in kidneys and intestines but not in other organs. We also performed cohabitation experiments to determine whether infections could be exchanged among species or only within species. Uninfected olive flounder and red sea bream, Pagrus major, cohabitating with emaciated olive flounder showed 100% and 0% cumulative mortality, respectively. Thus the cause of emaciation in cultured olive flounder of Korea is likely due to a new parasite.

Study on the Seed Production of Olive Flounder Paralichthys olivaceus in a Pilot Scale Recirculating Aquaculture System

  • Park, Jeong-Hwan;Jo, Jae-Yoon;Kim, Pyong-Kih
    • Journal of Aquaculture
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    • v.21 no.3
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    • pp.125-132
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    • 2008
  • Seed production of olive flounder Paralichthys olivaceus was performed in a pilot RAS. The growth of juvenile olive flounder and changes in water quality were monitored for the entire production period. The pilot RAS consisted of 8 circular culture tanks($4.0mD{\times}1.0mH$), 2 trickling biofilters($1.7mD{\times}2.0mH$), 2 protein skimmers ($0.8mD{\times}2.5mH$), and 4 sedimentation chambers($0.7mD{\times}1.5mH$). The culture surface area was about $100.5m^2$ and the actual working volume was about $106.9m^3$. As many as 300,000 fertilized olive flounder eggs were initially distributed into 2 culture tanks with the water temperature at $19.0^{\circ}C$. Live feeds such as rotifers and Artemia nauplii were fed until the 32nd day after hatching, and a commercial diet was fed from the 19th day to the end of the experiment. After 70 days, 150,256 juveniles with a body length of $65.8{\pm}3.9mm$ were produced in the RAS, with a daily growth rate for body length of 4.7%/day. At this time, the final culture density was 1,495 individuals $m^{-2}$, and 13.6 L of makeup water, 0.071 kW of electricity and 0.025 L of diesel fuel were used to produce a juvenile olive flounder. During metamorphosis of the larvae, the TAN concentration increased to 0.99 mg/L, which made the larvae sensitive to result in some mortality. However no more massive mortality occurred at the juvenile stage after metamorphosis even at a TAN concentration of 4.25 mg/L and a ${NO_2}^{-}-N$ concentration of 2.45 mg/L.