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Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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CYTOGENETIC ANALYSIS OF CHILDREN WITH AUTISM (자폐장애 환자의 세포유전학적 분석)

  • Jung, Chul-Ho;Lee, Je-Young;Park, Young-Nam;Park, Jong-Han;Kim, Jung-Bum;Kim, Jae-Ryong;Chun, Hyo-Jin
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.5 no.1
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    • pp.108-117
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    • 1994
  • Twenty nine children with autism and thirty children with mental retardation were examined for association between autism and chromosomal disorders including fragile X. The peripheral blood was cultured in Medium 199 with methotrexate and without methorexate for 70 hours. Thirty metaphase cells in each case were karyotyped in all samples. Chromosomal abnormalities were found in 11 cases(37.9%) of autistic disorder and 10 cases (33.3%) of mental retardation, but in none of fragile(X)(q27.3) from all cases. Chromosomal abnormalities were present on group A, C, D and X in autistic disorder and on group A, B, C, D, E and X in mental retardation. No specific chromosomal region was found in both autistic disorder and mental retardation. Types of chromosomal disorders were only fragile and/or gap but no numerical abnormality was present in all cases. Number of cells which revealed fragile sites were 31 cells(3.6%) out of 870 cells in autistic disorder and 29 cells(3.2%) out of 900 cells in mental retardation Number of cells which revealed gaps were 43 cells(4.9%) out of 870 cells in autistic disorder and 35 cells(3.9%) out of 900 cells in mental retardation. Autistic disorder may not be directly correlated with fragile X but with nonspecific chromosomal breakages from these data.

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Exposure Assessments of Environmental Contaminants in Ansim Briquette Fuel Complex, Daegu(II) - Concentration distribution and exposure characteristics of TSP, PM10, PM2.5, and heavy metals - (대구 안심연료단지 환경오염물질 노출 평가(II) - TSP, PM10, PM2.5 및 중금속 농도분포 및 노출특성 -)

  • Jung, Jong-Hyeon;Phee, Young-Gyu;Lee, Jun-Jung;Oh, In-Bo;Shon, Byung-Hyun;Lee, Hyung-Don;Yoon, Mi-Ra;Kim, Geun-Bae;Yu, Seung-do;Min, Young-Sun;Lee, Kwan;Lim, Hyun-Sul
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.3
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    • pp.380-391
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    • 2015
  • Objectives: The objective of this study is to assess airborne particulate matter pollution and its effect on health of residents living near Ansim Briquette Fuel Complex and its vicinities. Also, this study measured and analyzed the concentration of TSP, $PM_{10}$, $PM_{2.5}$, and heavy metals which influences on the environmental and respiratory disease in Ansim Briquette Fuel Complex, Daegu, Korea. Methods: In this study, we analyzed various environmental pollutants such as particulate matter and heavy metals from Ansim Briquette Fuel Complex that adversely affected local residents's health. In particular, we verified the concentration distribution and characteristics of exposure for TSP, $PM_{10}$, and $PM_{2.5}$ among particulate matters, and heavy metals(Cd, Cr, Cu, Mn, Ni, Pb, Fe, Zn, and Mg). In that regard, the official test method on air pollution in Korea for analysis of particulate matter and heavy metal in atmosphere were conducted. The large capacity air sampling method by the official test method on air pollution in Korea were applied for sampling of heavy metals in atmosphere. In addition, we evaluated the concentration of seasonal environmental pollutants for each point of residence in Ansim Briquette Fuel Complex and surrounding area. The sampling measured periods for air pollutants were from August 11, 2013 to February 21, 2014. Furthermore, we measured and analyzed the seasonal concentrations(summer, autumn and winter). Results: The average concentration for TSP, $PM_{10}$, and $PM_{2.5}$ by direct influence area at Ansim Briquette Fuel Complex were 1.7, 1.4 and 1.9 times higher than reference region. In analysis results of seasonal concentrations for particulate matter in four direct influence and reference area, concentration levels for winter were generally somewhat higher than concentrations for summer and autumn. The average concentrations for Cd, Cr, Mn, Ni, Pb, Fe, and Zn in direct influence area at Ansim Briquette Fuel Complex were $0.0008{\pm}0.0004{\mu}g/Sm^3$, $0.0141{\pm}0.0163{\mu}g/Sm^3$, $0.0248{\pm}0.0059{\mu}g/Sm^3$, $0.0026{\pm}0.0011{\mu}g/Sm^3$, $0.0272{\pm}0.0084{\mu}g/Sm^3$, $0.4855{\pm}0.1862{\mu}g/Sm^3$, and $0.3068{\pm}0.0631{\mu}g/Sm^3$, respectively. In particularly, the average concentrations for Cd, Cr, Mn, Ni, Pb, Fe, and Zn in direct influence area at Ansim Briquette Fuel Complex were 1.9, 3.6, 2.1, 1.9, 1.4, 2.6, and 1.2 times higher than reference area, respectively. The continuous monitoring and management were required for some heavy metals such as Cr and Ni. Moreover, the average concentration in winter for particulate matter in direct influence area at Ansim Briquette Fuel Complex were generally higher than concentrations in summer and autumn. Also, average concentrations for TSP, $PM_{10}$, and $PM_{2.5}$ were from 1.5 to 2.0 times, 1.2 to 1.8 times, and 1.1 to 2.3 times higher than reference area, respectively. In results for seasonal atmospheric environment, TSP, $PM_{10}$, $PM_{2.5}$, and heavy metal concentrations in direct influence area were higher than reference area. Especially, the concentrations in C station were a high level in comparison with other area. Conclusions: In the results, some particulate matters and heavy metals were relatively high concentration, in order to understand the environmental pollution level and health effect in surrounding area at Ansim Briquette Fuel Complex. The concentration of some heavy metals emitted from direct influence area at Ansim Briquette Fuel Complex were relatively higher than reference area. In particular, average concentration for heavy metals in this study were higher than average concentrations in air quality monitoring station for heavy metal for 7 years in Deagu metropolitan region. Especially, the residents near Ansim Briquette Fuel Complex may be exposed to the pollutants(TSP, $PM_{10}$, $PM_{2.5}$, and heavy metals, etc) emitted from the factories in Ansim Briquette Fuel Complex.

Quantitative Evaluation of the Corticospinal Tract Segmented by Using Co-registered Functional MRI and Diffusion Tensor Tractography (정상인에서 기능적 뇌 자기공명영상과 확산텐서영상 합성기법을 이용한 피질척수로의 위치에 따른 정량적 분석)

  • Jang, Sung-Ho;Hong, Ji-Heon;Byun, Woo-Mok;Hwang, Chang-Ho;Yang, Dong-Seok
    • Investigative Magnetic Resonance Imaging
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    • v.13 no.1
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    • pp.40-46
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    • 2009
  • Purpose : The purpose of this study was to investigate the quantitative evaluation of the corticospinal tract (CST) at the multiple levels by using functional MRI (fMRI) co-registered to diffusion tensor tractography (DTT). Materials and Methods : Ten normal subjects without any history of neurological disorder participated in this study. fMRI was performed at 1.5 T MR scanner using hand grasp-release movement paradigm. DTT was performed by using DtiStudio on the basis of fiber assignment continuous tracking algorithm (FACT). The seed region of interest (ROI) was drawn in the area of maximum fMRI activation during the motor task of hand grasp-release movement on a 2-D fractional anisotropy (FA) color map, and the target ROI was drawn in the cortiocospinal portion of anterior lower pons. We have drawn five ROIs for the measurement of FA and apparent diffusion coefficient (ADC) along the corona radiata (CR) down to the medulla. Results : The contralateral primary sensorimotor cortex (SM1) was mainly found to be activated in all subjects. DTT showed that tracts originated from SM1 and ran to the medulla along the known pathway of the CST. In all subjects, FA values of the CST were higher at the level of the midbrain and posterior limb of internal capsule (PLIC) than the level of others. Conclusion : Our study showed that co-registered fMRI and DTT has elucidated the state of CST on 3-D and analyzed the quantitative values of FA and ADC at the multiple levels. We conclude that co-registered fMRI and DTT may be applied as a useful tool for clarifying and investigating the state of CST in the patients with brain injury.

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Recombination and Expression of VP1 Gene of Infectious Pancreatic Necrosis Virus DRT Strain in a Baculovirus, Hyphantria cunea Nuclear Polyhedrosis Virus (전염성 췌장괴저바이러스 DRT Strain VP1유전자의 Baculovirus Hyphantria cunea Nuclear Polyhedrosis Virus에 재조합과 발현)

  • Lee, Hyung-Hoan;Chang, Jae-Hyeok;Chung, Hye-Kyung;Cha, Sung-Chul
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.239-255
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    • 1997
  • Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.

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State-based Peridynamic Modeling for Dynamic Fracture of Plane Stress (평면응력 문제의 상태 기반 페리다이나믹 동적파괴 해석 모델링)

  • Ha, Youn Doh
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.28 no.3
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    • pp.301-307
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    • 2015
  • A bond-based peridynamic model has been shown to be capable of analyzing many of dynamic brittle fracture phenomena. However, there have been issued limitations on handling constitutive models of various materials. Especially, it assumes bonds act independently of each other, so that Poisson's ratio for 3D model is fixed as 1/4 as well as taking only account the bond stretching results in a volume change not a shear change. In this paper a state-based peridynamic model of dynamic brittle fracture is presented. The state-based peridynamic model is a generalized peridynamic model that is able to directly use a constitutive model from the standard theory. It permits the response of a material at a point to depend collectively on the deformation of all bonds connected to the point. Thus, the volume and shear changes of the material can be reproduced by the state-based peridynamic theory. For a linearly elastic solid, a plane stress model is introduced and the damage model suitable for the state-based peridynamic model is discussed. Through a convergence study under decreasing the peridynamic nonlocal region($\delta$-convergence), the dynamic fracture model is verified. It is also shown that the state-based peridynamic model is reliable for modeling dynamic crack propagatoin.

Numerical Modeling of Flow Characteristics within the Hyporheic Zones in a Pool-riffle Sequences (여울-소 구조에서 지표수-지하수 혼합대의 흐름 특성 분석에 관한 수치모의 연구)

  • Lee, Du-Han;Kim, Young-Joo;Lee, Sam-Hee
    • Journal of Wetlands Research
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    • v.14 no.1
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    • pp.75-87
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    • 2012
  • Hyporheic zone is a region beneath and alongside a stream, river, or lake bed, where there is mixing of shallow groundwater and surfacewater. Hyporheic exchange controls a variety of physical, biogeochemical and thermal processes, and provides unique ecotones in a aquatic ecosystem. Field and experimental observations, and modeling studies indicate that hyporheic exchange is mainly in response to pressure gradients driven by the geomorphological features of stream beds. In the reach scale of a stream, pool-riffle structures dominate the exchange patterns. Flow over a pool-riffle sequence develops recirculation zones and stagnation points, and this flow structures make irregular pressure gradient which is driving force of the hyporheic exchange. In this study, 3 D hydro-dynamic model solves the Reynolds-averaged Navier-Stokes equations for the surface water and Darcy's Law and the continuity equation for ground water. The two sets of equations are coupled via the pressure distribution along the interface. Simulation results show that recirculation zones and stagnation points in the pool-riffle structures dominantly control the upwelling and downwelling patterns. With decrease of recirculation zones, length of donwelling zone formed in front of riffles is reduced and position of maximum downwelling point moves downward. The numerical simulation could successfully predict the behavior of hyporheic exchange and contribute the field study, river management and restoration.

Tri-branched tri-anchoring organic dye for Visible light-responsive dye-sensitized photoelectrochemical water-splitting cells (염료감응형 광전기화학 물분해 전지용 Tri-branched tri-anchoring organic dye 개발)

  • Park, Jeong-Hyun;Kim, Jae-Hong;Ahn, Kwang-Soon
    • 한국신재생에너지학회:학술대회논문집
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    • 2010.06a
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    • pp.87-87
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    • 2010
  • Photoelectrochemical (PEC) systems are promising methods of producing H2 gas using solar energy in an aqueous solution. The photoelectrochemical properties of numerous metal oxides have been studied. Among them, the PEC systems based on TiO2 have been extensively studied. However, the drawback of a PEC system with TiO2 is that only ultraviolet (UV) light can be absorbed because of its large band gap (3.2 - 3.4 eV). Two approaches have been introduced in order to use PEC cells in the visible light region. The first method includes doping impurities, such as nitrogen, into TiO2, and this technique has been extensively studied in an attempt to narrow the band gap. In comparison, research on the second method, which includes visible light water splitting in molecular photosystems, has been slow. Mallouk et al. recently developed electrochemical water-splitting cells using the Ru(II) complex as the visible light photosensitizer. the dye-sensitized PEC cell consisted of a dye-sensitized TiO2 layer, a Pt counter electrode, and an aqueous solution between them. Under a visible light (< 3 eV) illumination, only the dye molecule absorbed the light and became excited because TiO2 had the wide band gap. The light absorption of the dye was followed by the transfer of an electron from the excited state (S*) of the dye to the conduction band (CB) of TiO2 and its subsequent transfer to the transparent conducting oxide (TCO). The electrons moved through the wire to the Pt, where the water reduction (or H2 evolution) occurred. The oxidized dye molecules caused the water oxidation because their HOMO level was below the H2O/O2 level. Organic dyes have been developed as metal-free alternatives to the Ru(II) complexes because of their tunable optical and electronic properties and low-cost manufacturing. Recently, organic dye molecules containing multi-branched, multi-anchoring groups have received a great deal of interest. In this work, tri-branched tri-anchoring organic dyes (Dye 2) were designed and applied to visible light water-splitting cells based on dye-sensitized TiO2 electrodes. Dye 2 had a molecular structure containing one donor (D) and three acceptor (A) groups, and each ended with an anchoring functionality. In comparison, mono-anchoring dyes (Dye 1) were also synthesized. The PEC response of the Dye 2-sensitized TiO2 film was much better than the Dye 1-sensitized or unsensitized TiO2 films.

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Evaluation of Dose Distribution Using Gafchromic $EBT^{(R)}$ Film (Gafchromic $EBT^{(R)}$ 필름을 이용한 선량분포의 평가)

  • Kang, Se-Sik;Ko, Seong-Jin;Jang, Eun-Sung
    • Journal of radiological science and technology
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    • v.30 no.2
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    • pp.139-145
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    • 2007
  • Dose evaluation for small field such as stereotactic radiosurgery was performed using $Gafchromic^{(R)}$ EBT film. Every film which irradiated 6MV photon beam was scanned and obtained the optical density(OD) by flat bed scanner after 24 hours of irradiation. This study compared dose from diode in water and Gafchromic $EBT^{(R)}$ film in acrylic phantom to verify the reliability of the film, and to evaluate the SRS in clinical dose distributions from calculation and measurement in the region of virtual target in humanoid and cylindrical phantoms were compared. The Gafchromic $EBT^{(R)}$ film was found to be linear up to 9Gy. The $D_{max}$ for 6 MV was measured at 1.5 cm from the surface by both of diode and the film. As the depth is deeper, the error was measured within $2{\sim}3%$ at $10{\sim}20\;cm$ depth. Comparing between distribution from calculation and measurement, we found that there is 5% error at 90% isodose line. We found that given dose could be measured accurately by using the phantoms. It was feasible to use the Gafchromic $EBT^{(R)}$ film in quality assurance of SRS.

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Study on Mercury Contamination in Multimedia Environment in Lake So-Yang (소양호 다중매체 내 수은오염실태 조사)

  • Park, Sang-Young;Yoo, Seong;Shin, Hyung-Cheol;Kim, Seong-Rak;Kim, Pyung-Rae;Ahn, Myung-Chan;Han, Young-Ji
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.3
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    • pp.202-211
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    • 2011
  • In this study the seasonal and spatial pattern of mercury (Hg) concentrations in multimedia environments were investigated in Lake So-Yang. Total mercury (TM) in water column greatly enhanced as turbidity and particulate organic carbon (POC) increased due to the severe runoff, suggesting that most of Hg existed as particulate Hg in Lake Soyang. We also collected 22 species of fish in Lake Soyang and Han River, and the average total mercury in fish was 0.073 ppm, lower than both Korean (0.5 ppm) and EPA criteria (0.3 ppm). However when considering the amount of fish intake for Korean the Hg criterion in fish must be more stringent than current value, and the advisory for fish consumption should be made. Hg in fish considerably varied with sampling sites, having the highest in the upper region (Yang-gu) and the lowest in downstream (Han River). This spatial variation was possibly derived by "bloom dilution" associated with high phosphorus loading or elevated DOC concentrations, or both. Total mercury in sediment varied from 69.9 to $98.3{\mu}g/kg$, which was relatively lower than those measured in Voyageurs National Park in Minnesota in USA ($102{\sim}364{\mu}g/kg$).