• 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.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.06a
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• pp.345-350
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• 2005

Acute toxicities of five pharmaceutical products were evaluated with aquatic microbes, invertebrates, and fish. The test pharmaceuticals, i.e., cimetidine, carbamazepine, diltiazem, acetaminophene, and metformin have been often detected in aquatic environment, but theire cological hazard on receptors of various trophic levels has seldom been evaluated. In the present study, we conducted acute toxicity assays with a marine bacterium, Vibrio fischeri, an invertebrate, Daphnia magna, and a fish, Japanese medake (Oryzias latipes). In general, D. magna, showed the most sensitive response to the test chemicals. Diltiazem exhibited the lowest EC50 value after 96 hr of exposure at 7.6 mg/L, followed by cimetidine >acetaminophen > metformin = carbamazepine in an order of decreasing susceptibility. With the fish, diltiazem and carbamazepine showed the 96 hr EC50 values at 14.1${\sim}$35.4 mg/L while acetaminophen, cimetidine, and metformin did not cause 50% mortality at 100 mg/L. Similar pattern was noted with the Microtox Assay, with which the median effective concentrations for acetaminophen, cimetidine, and metformin were found at the range between 301.8 and 755.4 mg/L. Carbamazepine and diltiazem exposure to the microbes resulted in EC50 values around 50 mg/L. Predicted no effect concentrations (PECs) of these pharmaceuticals derived from the EC5O values obtained from this study, and predicted environmental concentrations (PECs) obtained from available literatures were utilized to estimate ecological risks of the test compounds. No test pharmaceuticals resulted in risk quotients (PEC/PNEC) greater than 1, which suggests no serious potential ecological concerns. It should be noted however that further studies including the refinement of PEC derivation, identification and toxicity assessment of the metabolites and/or their interactions with other stressors may be warranted to better understand the environmental consequences of the residual pharmaceutical discharge to the waterway.

• 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.

• Korean Journal of Ecology and Environment
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• v.38 no.3 s.113
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• pp.420-428
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• 2005

Effects of three biological control agents such as Xanthobacter autotrophycus, Tanichthys albonubes and Oryzias latipes on the morphology and growth of cyanobacterium Microcystis aeruginosa were studied. The experiments were consisted of six treatments of living organism (LO) and culture filtered water of three organisms (CFW). Three LOs effectively decreased the density of M. aeruginosa, and then cyanobacteria hardy showed in the microscopic field after 5 days of cultivation. All LO and CFW agents induced the colonial formation of cyanobacterium M. aeruginosa, although there were little differences in colony formation according to the kinds, density and type of treatment. In particular, the higher density treatment of fish CFW induced effectively the colony formation of cyanobacteria, compared to the bacterial LO and CFW. Thus, the application of bio agents to control the cyanobacterial bloom is needed to the further study to diminish the adverse effects such as the enhancement of colony formation towards on the new bloom against the aquatic ecosystem.

• 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.

• Journal of Korean Society on Water Environment
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• v.22 no.1
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• pp.37-44
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• 2006

Ecotoxicity assessments of 90 selected effluents of 22 industry types from 2002 to 2004 in Korea were evaluated by a toxic battery of bioassay test using fish Oryzias latipes, invertebrate Daphnia magna, algae Selenastrum capricornutum and bacteria Vibrio fischeri with the physicochemical measurement items and permit concentrations on the present Water Quality Conservation Act in Korea. Total toxic unit (${\Sigma}TU$) of 8 industry types of 22 industry types by the toxic battery appeared in order of the value site as follows; Pigment Dye Manufacturing (${\Sigma}TU$ 217.1) > Textile and Dye (${\Sigma}TU$ 39.3) > Semiconductor Electronic Manufacturing (Small) (${\Sigma}TU$ 25.6) > Wastewater and Sewage Treatment Plants (${\Sigma}TU$ 25.4) > Coating (${\Sigma}TU$ 23.8) > Leather Skin Manufacturing (${\Sigma}TU$ 18.0) > Synthetic Resin Manufacturing (${\Sigma}TU$ 15.6) > Assemble Metal Manufacturing (${\Sigma}TU$ 10.7). Our results demonstrate that ecotoxicity assessment, by bioassay test, is effective and practical for industrial wastewater management for 90 selected effluents with the limitation of the physicochemical permit. Among 90 effluents, 9 samples failed physicochemical permit limitation and 81 passed it. In result of ecotoxicity assessment of 90 effluents by the toxic battery, 76 effluents exhibited ecotoxicity and the others did not. The physicochemical measurement items and permit concentrations on the present Water Quality Conservation Act in Korea were low related to the ecotoxicity value by the toxic battery and appeared limited for water quality management to water-ecosystem and environment-friendly management of water.

• Environmental Analysis Health and Toxicology
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• v.9 no.3_4
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• pp.91-101
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• 1994

In order to evaluate the ecotoxicological harzardousness of synthetic surfactants on Han river, Jung-ryang and Jin-Wi stream, we used the Ecotoxicological Risk Quotient (ERQ). The chemical harzardousness is evaluated by the balance of the toxicity and concentration in the environment.. Then, ERQ is defined as follows; ERQ = - log ( Concentration in the environment / Effective concentration in the test ) ERQ of chemical is a logtrighmic value of ratio of a chemical concentration and the toxicity in the laboratory. In case of small ERQ, tie chemical harzardousness is high. If ERQ equals O, the same biological effect as in the laboratory test will be observed in the enviromment by the chemicals. ERQ values of the chemicals were calculated using the maximum concentration in water environment which were cited from the annual report by our ministry of environment, and EC$_{50}$ of Daphnia magna (water flea; acute immobilization test) LC$_{50}$ of Oryzias latipes (fish; acue toxicity test) and EC$_{50}$ of chlorella vulgaris (alga; growth inhibition test), which were taken from the annual report of "Chemical in environment" by Japan EA. Liner alkylbenzene sulfonate (determined to MBAS) showed the high average values with more than 2.0 to three species in Han river and Jin-wi stream, and these results mean to be favorable to environmental safety. The areas of Jung-ryang stream were polluted, as the average values of ERQ were less than 2.0 with equal to three species, and attention should be paid. Therefore, they must be inspected again because their concentration in the environment may have changed during that period. The chemical harzardousness can be numerated with ERQ, and it can be a help to find the chemicals that should be kept under observation and to see whether the chemical pollution is improved or worsened. The determination of the chemical concentration in the environment and toxicity are essential for the effective use of ERQ.se of ERQ.

• Journal of The Korean Society of Agricultural Engineers
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• v.46 no.3
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• pp.93-105
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• 2004

The Shihwa wetland, a newly developed constructed wetland prototype in Korea with an area of approximately 104 ha, was established to improve the water quality severely polluted inflow streams into Lake Shihwa. Because this wetland could play a role as an ecological park in addition to improving the water quality, an ecological impact of the Shihwa constructed wetland has been a national concern. This paper focused on reporting the survey results for fauna among the entire investigation results for 3 years. A total of 129 terrestrial insect species have been observed from August 2001 to June 2002. Among them, Ischnura asiatica (Brauer) (order Odonata), Scymnus species (order Coleoptera) and Orthopteran species were frequently found in the reed bushes. A total of 77 bird species were recorded in a seasonal count, the maximum number of species was 34 in winter and the maximum number of individuals was 4,599 in summer. For the freshwater fish, only 4 species were found in 2000, however in 2001 and 2002, 12 species and 459 individuals were collected at four survey points. Among these 12 species, the dominant species were Mugil cephalus(36%), followed by Carassius auratus (25%) and Rhinogobius brunneus (22%). Meanwhile,12 individuals of Oryzias latipes were observed nearby, mostly downstream of the wetland. For the Herpetofauna at four survey areas, 3 species of amphibians and 3 species of reptiles were recorded. Because of remaining salinity in the soil of the Shiwha constructed wetland, Herpetofauna inflow to the wetland was scanty and mainly inhabited the upstream area. A total of 8 mammal species were recorded. Small-sized species were the striped field mouse, the Ussurian harvest-mouse, the Manchurian reed vole and the brown rat. Middle- and large-sized species were Korean water-deer, Korean raccoon dogs, Korean yellow weasels and feral cats.

• Toxicological Research
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• v.18 no.1
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• pp.13-22
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• 2002

In Korea, 2,320 tonnes of acetanilide were mostly wed as intermediates for synthesis in phar-maceuticals or additives in synthesizing hydrogen peroxide, varnishes, polymers and rubber. Only small amount of 120 kg were wed as a stabilizer for hydrogen peroxide solution for hair colouring agents in 1998. Readily available environmental or human exposure data do not exist in Korea at the present time. However, potential human exposures from drinking water, food, ambient water and in work places are expected to be negligible because this chemical is produced in the closed system in only one company in Korea and the processing factory is equipped with local ventilation and air filtering system. Acetanilide could be distributed mainly to water based on EQC model. This substance is readily biodegradable and its bioaccumulation is low. Acute toxicity of acetanilide is low since the L $D_{50}$ of oral exposure in rats is 1,959 mg/kg bw. The chemical is not irritating to skin, but slightly irritating to the eyes of rabbits. horn repeated dose toxicity, the adverse effects in rats were red pulp hyperplasia of spleen, bone marrow hyperplasia of femur and decreased hemoglobin, hematocrit and mean corpuscular hemoglobin concentration. The LOAEL for repeated dose toxicity in rats was 22 mg/kg/day for both sexes. Acetanilide is not considered to be genotoxic. In a reproductive/developmental toxicity study, no treatment-related changes in precoital time and rate of copulation, impregnation, pregnancy were shown in all treated groups. The NOAELs for reproduction and developmental toxicity (off-spring toxicity) are considered to be 200 mg/kg bw/day and 67 mg/kg bw/day, respectively. Ecotoxicity data has been generated in a limited number of aquatic species of algae (72 hr- $E_{b}$ $C_{50}$; 13.5 mg/l), daphnid (48hr-E $C_{50}$ > 100 mg/l) and fish (Oryzias latipes, 96hr-L $C_{50}$; 100 mg/l). Form the acute toxicity values, the predicted no effect concentration (PNEC) of 0.135 mg/1 was derived win an assessment factor of 100. On the basis of these data, acetanilide was suggested as currently of low priority for further post-SIDS work in OECD.in OECD.D.

• 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.