• Journal of Environmental Health Sciences
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• v.28 no.2
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• pp.131-139
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• 2002

초기생애단계의 일본송사리를 실험어류로 사용하여 TCDD와 PCB 126의 생물농축정도를 비교 측정하였고, TCDD와 PCB 126의 체외 배설에 대한 동태학적 특성을 비교 분석하였다. 생물농축실험은 초기생애단계의 새끼치어 및 어린 물고기에 TCDD와 PCB 126을 여러 농도로 물에 용해시켜 96시간 동안 정체된 상태로 폭로시킨 후 생물농축계수(BCF)와 지방-표준화한 생물농축계수(BCF$_{L}$)를 측정하였다. TCDD와 PCB 126의 일본송사리 초기생애단계의 새끼치어 및 어린 물고기에서 측정된 96-h BCF값 및 96-h BCF$_{L}$값은 폭로된 농도와 상관없이 비슷하게 나타나 반면, 생체크기가 클수록 그 값은 작게 나타났다. 일본송사리 새끼치어, 어린 물고기, 성어의 생체 내 총 지방함량은 각각 5.7, 4.2, 4.6%로 측정되었다. 체외배설 동태실험은 어린 물고기에 TCDD와 PCB 126을 3가지 농도롤 물에 용해시켜 96시간 동안 정체된 상태로 폭로시킨 후, flow-through장치로 공급되는 맑은 물로 옮겨 42일간 사양하면서 0, 7, 14, 21, 28, 42일에 생체 내 TCDD와 PCB 126 잔류량을 측정하여 분석하였다. 배설은 1차 반응속도론적으로 이루어졌으며 한 구획모형보다 두 구획모형이 더 적합한 것으로 분석되었다. 두 구획모형을 사용한 분석에서 TCDD와 PCB 126의 체외배설 반감기(t$\frac{1}{2}$) 및 배설속도상수($\beta$)는 각각 27.2일과 32.3일 및 0.025/d와 0.021/d로 측정되었다.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• v.3 no.1
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• pp.47-53
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• 2022

The escape behavior of prey fish to predator attack is directly linked to the survival of the fish. In this study, I explored the escape behavior of Medaka fish to bird attacks. To simulate the attack, I designed a model triangular-shaped bird to slide along a fishing line connected between rods at both ends of the tank. The triangular shape was set to 10×15 (S=1), 15×20 (S=2), and 20×25 cm (S=3) with base×height. The slope (θ) of the fishing line, which determines the attack speed of the model bird, was set to values of 15° (θ=1), 30° (θ=2), and 45° (θ=3). The escape behavior was characterized using five variables: escape speed (ν), escape acceleration (α), responsiveness (γ), branch length similarity entropy (ε), and alignment (ϕ). The experimental results showed when (S, θ)=(fixed, varied), the change in values of the five variables were not significant. Thus, the fish respond more sensitively to S than to θ In contrast, when (S, θ)=(varied, fixed), ν, α, and γ showed increasing trends but ε and ϕ did not change much. This indicates the nature of fish escape behavior irrespective of the threat is inherent in ε and ϕ. I found that fish escape behavior can be divided into two types for the five physical quantities. In particular, the analysis showed that the type was mainly determined by the size of the model bird.

• Environmental Analysis Health and Toxicology
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• v.24 no.4
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• pp.321-332
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• 2009

본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

• Environmental Analysis Health and Toxicology
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• v.18 no.2
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• pp.145-153
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• 2003

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals. Japanese medaka was treated by copper of appropriate sublethal concentrations after starvation for 48 hr. The untreated individuals showed common behavioral characteristics (i.e. , smooth and linear movements). Locomotive activity of the fish was monitored using an image processing and automatic data acquisition system. When treated with copper (100 ppb), the fish showed shaking patterns more frequently. As the concentration of copper increased to 1,000 ppb, activity decreated, and the fish showed an erratic movement. Fish were exposed to copper at various concentrations (0,100 and 1,000 ppb) for 24 hrs, and acetylcholine esterase (AChE) activity was observed. When fish were exposed to 1,000 ppb of copper, the body AChE activities appeared to decrease but the head AChE activities showed little change. Expressions of tyrosine hydroxylase (TH) protein in the different organs from both head (brain) and body (kidney) portions affected by the copper treatment were analyzed using immunohistochemical technique compared with control. Five organs of the fish (olfactory bulb, hyothalamus, optic lobe, pons and myelencephalon regions) showed a relatively strong TH protein expression in the control experiment. A differential expression of TH, however, was observed in the treatment (100 ppb and 1,000 ppb). The treatment (1,000 ppb) significantly suppressed TH protein production in the brain regions. In kidney, however, the same treatment caused little suppression compared with the control. Copper appeared to be less effective in suppression of TH than diazinon, a known TH suppressor. It was concluded that TH could be used at a potential biomarker to monitor the acute copper toxicity in Japanese medaka.

• The Korean Journal of Pesticide Science
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• v.18 no.1
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• pp.8-13
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• 2014

This study evaluated in vitro cytotoxicity of 5 pesticides, including 2 herbicides, 2 germicides, and an insecticide, as an alternative to the fish acute toxicity test. The in vitro cytotoxicity was tested using a neutral red uptake (NRU) assay with epithelioma papulosum cyprini (EPC) cells that originated from the epidermal tissue of Cyprinus carpio (common carp). An in vivo fish acute toxicity test was conducted according to OECD Test Guideline No. 203 using Aphyocypris chinensis (Chinese bleak), Oryzias latipes (Japanese medaka), and C. carpio. The results showed that the sensitivity of the cell viability assay for the pesticides was similar to the fish acute test in ranking order despite having approximately 10 times less absolute sensitivity. The $r^2$ correlation values were calculated as 0.38 (p = 0.26), 0.76 (p = 0.05) and 0.90 (p = 0.01) for A. chinensis, O. latipes, and C. carpio, respectively. These results suggested that the potential of EPC cell viability assay as an alternative to the fish acute toxicity test due to their good correlation and NRU assay is expected to serve as a useful tool for predicting acute fish lethality for pesticides if further studies with a large set of pesticides are conducted.

• Environmental Analysis Health and Toxicology
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• v.22 no.4
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• pp.305-312
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• 2007

환경오염을 신속하게 모니터링하기 위한 생물지표의 개발은 증가하고 있는 오염의 심각성에 비추어 매우 중요한 과제로 여겨지고 있다. 본 연구에서는 독성물질처리에 의하여 선택적으로 발현이 조절되는 단백질의 동정을 통하여 독성물질에 대한 단백질 생물지표를 발굴하고자 시도하였다. 즉, 송사리(Oryzias latipes)를 유기인계 살충제인 다이아지논(diazinon)에 0, 0.1, 1, 5 mg/L 농도로 24시간 노출시킨 후, 머리와 몸통부분으로 나누어 단백질 발현패턴을 분석하였다. 본 시스템에서 다이아지논 처리에 의하여 유의적으로 발현이 증가된 단백질로서 alpha-tubulin, ribonuclease pancreatic precursor, protein hfq 등을 동정하였으며, 이 가운데 alpha-tubulin과 $hsp90{\beta}$의 발현이 다이아지논 농도에 의존적으로 증가하는 것을 semi-quantitative RT-PCR방법으로 확인하였다. 이와 같이 다이아지논 처리에 특이적으로 발현이 증가된 송사리 단백질들은 노출평가를 위한 생물지표로서 개발에 응용될 수 있을 것으로 평가된다.

• Korean Journal of Ecology and Environment
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• v.40 no.2
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• pp.173-183
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• 2007

The Ministry of Environment plans to introduce the Whole Effluent Toxicity (WET) system in Korea. The WET test is well established in developed countries with recognizing of the limitations of physicochemical analysis method and potential risk of chemicals in water medium. Therefore, it is essential to build the ecotoxicity infrastructure for the induction of WET test. In this study, we extensively collected the domestic and foreign toxicity test methods which employ native test species to Korea. And we suggested that the domestic ecotoxicity test methods with domestic test species in Korea through extracting the range of test conditions. Five domestic fish species selected were Carassiu auratus (Crucian carp), Cyprinus carpio(Common carp), Gasterosteus aculeatus (Three spine stickleback), Misgurnus anguillicaudatus (Oriental weather fish) and Oryzias latipes (Japanese medaka), The toxicity test methods with native test species to Korea were collected from the standard methods (OECD, U.S. EPA, ASTM), government reports, SCI papers and domestic papers. We collected the 32 test methods, and suggested the suitable aquatic toxicity test methods for fish. It is expected that this study could prove a useful information to establish the ecotoxicity test methods with domestic aquatic organisms in Korea henceforth.

• Environmental Analysis Health and Toxicology
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• v.22 no.2 s.57
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• pp.171-175
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• 2007

Indigenous species means a species that is likely, due to historical presence, to occur at a specified site for some portion of its life span. Therefore, indigenous species can be useful as an indicator to assess environmental risk caused by hazardous chemicals in a specific site. So far a few toxicity studies using freshwater species which are indigenous to Korea have been carried out. In this study, a freshwater shrimp (Neocardina denticulata) indigenous to Korea was used for acute toxicity test of heavy metals. Neocardina denticulata were exposed to cadmium chloride $(CdCl_2)$, copper chloride $(CuCl_2)$ and zinc chloride $(ZnCl_2)$ using automatic flow-through system for 96 hours. The 96h LC50s were calculated as 0.043 $(0.042{\sim}0.045)mg\;CdCl_2/L,\;0.104(0.098{\sim}0.113)mg\; CuCl_2/L\;and\;2.021\;(1.633{\sim}2.594)mg\;ZnCl_2/L$. When compaired with some international standard species such as medaka(Oryzias latipes), Neocardina denticulata had high sensitivity. Therefore, this study suggested that Neocardina denticulata have possibilities for a sensitive test species to test heavy metal toxicity in aqua-system.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2002.10a
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• pp.156-156
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• 2002

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.

• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.266-273
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• 2007

Heat shock proteins (HSPs) are a family of highly conserved proteins playing an important role in the functioning of unstressed and stressed cells. The HSP70 family, the most widely studied of the hsps, is constitutively expressed (hsc70) in unstressed cells and is also induced in response to stressors (hsp70), especially those affecting the protein machinery. The HSP/HSC70 proteins act as molecular chaperones and are crucial for protein functioning, including folding, intracellular localization, regulation, secretion, and protein degradation. Here, we report the identification and characterization of the putative amino acid sequence deduced from one cDNA clone identified as heat shock protein 70. The alignment showed that the putative sequence is 100% identical to the heat shock protein 70 cognate (HSC 70) of olive flounder. The 5'-flanking region sequence (approximately 1 kb) ahead of the hsc70 gene was cloned by genome walking and a putative core promoter region and transcription elements were identified. We characterized the promoter of the olive flounder hsc70 gene by examining the ability of 5'-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos.