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Expression Pattern of Early Growth Response Gene 1 during Olive Flounder (Paralichthys olivaceus) Embryonic Development

  • Yang, Hyun;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Kim, Kyung-Kil
    • Development and Reproduction
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    • v.18 no.4
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    • pp.233-240
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    • 2014
  • The early growth response protein 1 (Egr-1) is a widely reported zinc finger protein and a well known transcription factor encoded by the Egr-1 gene, which plays key roles in many aspects of vertebrate embryogenesis and in adult vertebrates. The Egr-1 expression is important in the formation of the gill vascular system in flounders, which develops during the post-hatching phase and is essential for survival during the juvenile period. However, the complete details of Egr-1 expression during embryo development in olive flounder are not available. We assessed the expression patterns of Egr-1 during the early development of olive flounders by using reverse transcription polymerase chain reaction (RT-PCR) analysis. Microscopic observations showed that gill filament formation corresponded with the Egr-1 expression. Thus, we showed that Egr-1 plays a vital role in angiogenesis in the gill filaments during embryogenesis. Further, Egr-1 expression was found to be strong at 5 days after hatching (DAH), in the development of the gill vascular system, and this strong expression level was maintained throughout all the development stages. Our findings have important implications with respect to the biological role of Egr-1 and evolution of the first respiratory blood vessels in the gills of olive flounder. Further studies are required to elucidate the Egr-1-mediated stress response and to decipher the functional role of Egr-1 in developmental stages.

Effects of acetaminophen administration on liver histopathology, serum GOT/GPT levels and circulating microRNA-122 concentration in olive flounder (Paralichthys olivaceus)

  • Najib, Abdellaoui;Kim, Dong Soo;Kim, Ki Hong
    • Journal of fish pathology
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    • v.29 no.1
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    • pp.25-33
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    • 2016
  • In human medicine, circulating microRNAs have been successfully utilized as early biomarkers for various abnormalities and disease states. Vertebrate miR-122 is a liver-specific microRNA which is expressed almost solely in hepatocytes and plays an important role in the regulation of hepatocyte function. In this study, to evaluate the potential utility of circulating miR-122 as a biomarker for liver injury in olive flounder (Paralichthys olivaceus), fish were orally intubated with two doses of acetaminophen (500 mg/kg or 1.000 mg/kg of body weight), and the expression of miR-122 in serum was quantified using real time-PCR. Histological change in liver, and the enzymatic activity of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were also analyzed. The results showed that miR-122 was higher in acetaminophen administered groups compared to control group. The histopathological effect of acetaminophen on olive flounder liver was not distinct. The serum level of GPT and GOT was increased within 2 folds compared to control group by acetaminophen administration. However, the serum miR-122 level was increased more than 3 or 4 folds compared to the control group by administration of 1000 mg/kg of acetaminophen. These results suggest the possible use of miR-122 as an indicator of liver injury in olive flounder, even when histopathological effects are not distinctive.

Comparison of Extruded and Moist Pellets for Whole-body Proximate Composition and Growth Performance of Olive Flounder Paralichthys olivaceus (배합사료 및 습사료 공급에 따른 넙치(Paralichthys olivaceus)의 성장 및 어체성분 비교)

  • Kim, Kang-Woong;Lee, Jin-Hyuck;Bae, Ki-Min;Kim, Kyoung-Duck;Lee, Bong-Joo;Han, Hyon-Sob;Kim, Sung-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.6
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    • pp.810-817
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    • 2014
  • Two consecutive feeding trials were conducted to evaluate the effects of extruded pellet (EP) and raw fish-based moist pellet (MP) in the juvenile (experiment I) and sub-adult (experiment II) stages of olive flounder, Paralichthys olivaceus. The fish were distributed randomly to three aquarium tanks, as a group of 1,200 fish (initial mean weight $13.5{\pm}1.76g$) in experiment I, and as a group of 390 fish (initial mean weight $385{\pm}15.3g$) in experiment II. In experiment I, the weight gain, specific growth rate, feed efficiency, protein efficiency ratio, and survival of fish fed EP were all significantly higher than those of fish fed MP. In experiment II, no significant differences were observed in weight gain, specific growth rate and survival between the EP and MP groups. However, the feed efficiency and protein efficiency ratios of fish fed EP were significantly higher than those of fish fed MP. The results of this study indicate that EP could be developed to replace MP for market size production of olive flounder without any adverse effects on the growth performance. The dietary formulation used in this study could be used as an appropriate feed for olive flounder.

Survival and Growth in Larval Stage of Induced Hybrid between Olive Flounder (Paralichthys olivaceus) and Spotted Flounder (Verasper variegatus) (유도된 넙치와 범가자미간 잡종 자어의 생존 및 성장에 관한 연구)

  • Kim Kyung-Kil;Bang In Chul;Kim Yoon;Kim Dong Soo
    • Journal of Aquaculture
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    • v.8 no.1
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    • pp.69-76
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    • 1995
  • Hybrid fish were induced between olive flounder (Paralichthys olivaceus) female and spotted flounder (Verasper variegatus) male by artificial inseminations. Survival, growth and morphological changes in larval stage of hybirds were examined and compared to their parental species. Survival and growth up to 25 days after hatching of the hybrids were more similar to those of olive flounder than to those of spotted flounder (P<0.05). The ratio of yolk length to yolk width in hybrids was similar to that of female parents (P<0.05). The oil globule were resorbed within 18 days after hatching in hybrids and 14 days after hatching in olive flounder, and spotted flounder has no oil globule. These results indicated that survival, growth and morphological changes in larval stage of hybrids were similar to those of maternal olive flounder.

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Random Amplified Polymorphic DNA Analysis for Origin Identification of Olive Flounder (Paralichthys olivaceus) and Redlip Croaker (Pseudosciaena polyactis) (넙치와 조기의 원산지 판별을 위한 random amplified polymorphic DNA 패턴 연구)

  • Kang Duk-Jin;Lee Suk-Keun;Jin Deuk-Hee;Choi Suk-Jung
    • Journal of Life Science
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    • v.16 no.1
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    • pp.88-94
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    • 2006
  • The random amplified polymorphic DNA (RAPD) technique was investigated as a potential tool for the origin identification of olive flounder (Paralichthys olivaceus) and redlip croaker (Pseudosciaena polyactis). Olive flounder specimens were collected from North Korea and several locations of South Korea (Jumunjin, Tongyoung and Geoje). Fishes obtained from Tongyeong and Geoje were cultured products. Redlip croaker specimens were collected from South Korea and China. Consistent and distinct diagnostic bands were easily identified in the RAPD patterns of the olive flounder specimens. Although consistent diagnostic bands rarely appeared in the RAPD pattern of redlip croaker specimens because of their genetic heterogeneity, we were able to find potential diagnostic bands in the average RAPD pattern of each origin.

Removal of Organic Load from Olive Washing Water by an Aerated Submerged Biofilter and Profiling of the Bacterial Community Involved in the Process

  • Pozo, Clementina;Rodelas, Belen;Martinez-Toledo, M. Victoria;Vilchez, Ramiro;Gonzalez-Lopez, Jesus
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.784-791
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    • 2007
  • The present work aims to use a biofilter technology(aerated submerged filters) for the aerobic transformation at laboratory-scale of olive washing water(OWW) generated in the first steps of olive oil processing, as well as the genetic profiling and identification to the species level of the bacteria involved in the formation of the biofilm, by means of TGGE. Chemical parameters, such as biological oxygen demand at five days($BOD_5$) and chemical oxygen demand(COD), decreased markedly(up to 90 and 85%, respectively) by the biological treatment, and the efficiency of the process was significantly affected by aeration and inlet flow rates. The total polyphenol content of inlet OWW was only moderately reduced(around 50% decrease of the inlet content) after the biofilter treatment, under the conditions tested. Partial 16S rRNA genes were amplified using total DNA extracted from the biofilm and separated by TGGE. Sequences of isolated bands were mostly affiliated to the $\alpha-subclass$ of Proteobacteria, and often branched in the periphery of bacteria] genera commonly present in soil(Rhizobium, Reichenowia, Agrobacterium, and Sphingomonas). The data obtained by the experimentation at laboratory scale provided results that support the suitability of the submerged filter technology for the treatment of olive washing waters with the purpose of its reutilization.

Development of a Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Nervous Necrosis Virus in Olive Flounder Paralichthys olivaceus

  • Suebsing, Rungkarn;Oh, Myung-Joo;Kim, Jeong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.1021-1028
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    • 2012
  • In this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid, sensitive, and inexpensive detection of nervous necrosis virus (NNV) in olive flounder, Paralichthys olivaceus, in Korea. A set of six specific primers was designed to target the RNA 2 gene encoding the coat protein of Korean NNV strains. The RT-LAMP reaction successfully detected NNV after 30 min at $65^{\circ}C$. When the sensitivities among RT-LAMP, RT-PCR, and nested RTPCR were compared, the RT-LAMP was shown to be able to detect the RNA template at $2.58{\times}10^{-2}\;TCID_{50}/ml$, whereas the RT-PCR and nested RT-PCR were only able to detect the RNA template at $2.58{\times}10^2\;TCID_{50}/ml$ and $2.58TCID_{50}/ml$, respectively. Thus, the sensitivity of the RT-LAMP assay was higher than those of the RT-PCR assays. In the specificity test of the RT-LAMP, 2 genotypes of NNVs (SJNNV and RGNNV) were positive; however, no other fish viruses were positive with the primers, indicating that the RT-LAMP assay is only specific to NNV. A total of 102 olive flounder were collected from hatcheries between 2009 and 2011. The occurrence of NNV in olive flounder was determined to be 53.9% (55/102) by the RT-LAMP. On the other hand, the prevalence based on the nested RT-PCR and RT-PCR results was 33.8% (34/102) and 20.6% (21/102), respectively. This result indicates that the RT-LAMP assay developed in this study is suitable for early field diagnosis of NNV with high sensitivity.

Effects of various concentrations of garlic powder and garlic extract in the diets on growth, serum chemistry and immune response of juvenile olive flounder Paralichthys olivaceus

  • Cho, Sung-Hwoan;Lee, Sang-Min;Kwon, Mun-Gyeong
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.409-420
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    • 2010
  • Effects of various concentrations of garlic powder and garlic extract in the diets on growth, serum chemistry and immune response of olive flounder were determined. Thirty-five juvenile fish averaging 5.1 g were randomly distributed into 21 of 180 L flow-through tanks. Seven experimental diets with various concentrations of garlic powder (GP) and garlic extract (GE) were prepared in triplicate: GP-0 without garlic supplementation, GP-0.5, GP-1, GP-2, GP-3 and GP-5 diets containing garlic powder at the concentrations of 0.5, 1, 2, 3 and 5%, respectively at the expense of wheat flour and finally, GE-0.4 diet containing 0.4% garlic extract were prepared. At the end of the 8-week feeding trial, serum chemistry of fish was measured. In addition, twenty fish from each tank were artificially infected with E. tarda for the following 96 h to monitor cumulative mortality. Weight gain of fish fed GP-0 diet was higher than that of fish fed GP-1, GP-2, GP-3 and GP-5 diets. No difference in serum criteria (total protein, glucose, glutamate oxaloacetate transaminase, cholesterol and triglyceride levels) of olive flounder was found among the experimental diets except for glutamate pyruvate transaminase. Lysozyme activity of fish fed GP-0, GP-1, GP-3 and GE-0.4 diets was higher than that of fish fed GP-5 diet. The highest cumulative mortality was 93.3% in fish fed GP-0 diet at 96 h after E. tarda infection, followed by GP-3, GP-1, GP-5, GP-2, GP-0.5 and GE-0.4 diets. In considering these results, dietary inclusion of garlic powder and garlic extract has no distinctive positive effect on improvement in growth, serum chemistry and immune response of olive flounder in this experimental conditions, therefore, its application should be carefully considered.

Analysis of Manifestation of CC and CXC Chemokine Genes in Olive Flounders (Paralichthys olivaceus) Artificially Infected with VHSV during the Early Developmental Stage

  • Kim, Kyung-Hee;Kim, Woo-Jin;Park, Choul-Ji;Park, Jong-Won;Noh, Gyeong Eon;Lee, Seunghyung;Lee, Young Mee;Kim, Hyun Chul
    • Development and Reproduction
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    • v.22 no.4
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    • pp.341-350
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    • 2018
  • Chemokines is a small protein that plays a major role in inflammatory reactions and viral infections as a chemotactic factor of cytokines involved in innate immunity. Most of the chemokines belong to the chemokine groups CC and CXC. To investigate the immune system of the olive flounder (Paralichthys olivaceus), an expression pattern specifically induced in the early developmental stages of analysis is examined using qRT-PCR. We also examined tissue-specific expression of both CC and CXC chemokine in healthy olive flounder samples. CC and CXC chemokine shows increased expression after immune-related organs are formed compared to expression during early development. CC chemokine was more highly expressed in the fin, but CXC chemokine showed higher expression in the gills, spleen, intestines, and stomach. Spatial and temporal expression analysis of CC and CXC chemokine were performed following viral hemorrhagic septicemia virus (VHSV) infection. CC chemokine showed high expression in the gills, which are respiratory organs, whereas CXC chemokine was more highly expressed in the kidneys, an immune-related organ. These results suggest that CC and CXC chemokine play an important role in the immune response of the olive flounder, and may be used as basic data for the immunological activity and gene analysis of it as well as other fish.

Evaluation of Diacylglycerol as an Alternative to Dietary Fish Oil in Diets for Juvenile Olive Flounder Paralichthys olivaceus (넙치(Paralichthys olivaceus) 사료 내 어유(Fish oil) 대체원으로써의 diacylglycerol 이용성 평가)

  • Oh, Dae-Han;Kim, Min-Gi;Yun, Kawn-Sik;Lee, Kyeong-Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.4
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    • pp.366-373
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    • 2019
  • The objective of this study was to investigate the utilization of diacylglycerol (DAG) as a new dietary ingredient replacing fish oil in feed for juvenile olive flounder Paralichthys olivaceus. Fish oil based control diet (CON) was prepared and four other diets were formulated by replacing 50% of the fish oil in CON with one of five DAG: DAGL (1,3-lauryl glycerol) or DAGP (1,3-palmityl glycerol) in low or high concentrations (designated as DAGLL, DAGLH, DAGPL and DAGPH). Another diet was prepared replacing 100% of the fish oil in CON with a 1:1 mixture (DAGLP) of DAGL and DAGP. Olive flounder (13.4 g) were fed to apparent satiation, twice a day, for 12 weeks. Following the feeding trials, no significant differences were observed in growth performance, blood parameters and non-specific immune responses between CON and any of the DAG groups. Polyunsaturated fatty acid levels were not significantly affected by the inclusion of DAGs. Thus, DAGL or DAGP could be used to replace up to 50% of fish oil in fish feed without reducing growth performance, health or innate immunity. The replacement of up to 100% of dietary fish oil in olive flounder feed by DAGLP is also feasible.