• Environmental Analysis Health and Toxicology
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• v.22 no.3
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• pp.203-209
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• 2007

생활하수, 공장폐수, 농경유출수에 의해 수생태계로 유입된 다양한 화학물질들은 수서곤충이나 어류와 같은 수생생물에게 나쁜 영향을 주곤 한다. 비스페놀A와 노닐페놀을 포함하는 많은 화학물질들이 내분비계 장애물질(EDCs)로 의심되고 있고, 그들은 환경속에서 서로 다른 혼합형태로 공존하기도 한다. 따라서 비스페놀A와 노닐페놀의 혼합물이 독성과 생식학적 반응에 미치는 영향을 살펴보기 위해 일본산 송사리의 수정란 치사율, 부화율 및 부화시간, 치어의 성장율 및 비텔로제닌 농도 등이 측정되었다. 수정된 지 24시간 이내의 수정란을 대조군, 양성대조군($17{\beta}-estradiol$), 그리고 서로 다른 농도의 비스페놀A와 노닐페놀의 혼합물에 부화 후 60일까지 유수식 조건하에 노출시켰다. 수정란${\sim}$치자어 단계에서는 대조군과 비교하여 실험군의 치사율 및 부화율, 부화시간에 차이가 나타나지 않았으며, 부화 후 60일간의 노출 후 성장(길이, 무게)에 있어서도 비록 양성대조군에서 낮은 성장상태를 보였지만 다른 혼합물의 실험군들과는 차이를 보이지 않았다. 한편 체내 비텔로제닌 농도는 혼합물의 농도증가에 따라 증가하였으며 수컷의 경우 최저농도의 혼합물(Treatment A)을 제외한 실험군에서 농도증가에 따라 증가하였다. 반면 양성대조군의 경우 수컷이 발견되지 않았고 암컷 체내의 비텔로제닌 농도는 최고농도의 혼합물(Treatment D) 실험군과 비슷한 경향을 보였다. 위 실험을 통해 각각의 내분비계 장애물질이 개별적으론 생식발달 및 비텔로제닌 유도에 무영향농도(NOEC)라 하더라도 혼합된 경우 영향이 나타날 수 있다는 것을 보여주었으며, 이는 수환경 내 다양한 화학물들의 혼합효과(combination effect)가 생태위해성평가를 좀더 면밀하게 하기 위해서 주의 깊게 고려되어야 한다고 제안한다.

• 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.

• Korean Journal of Environmental Biology
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• v.25 no.1
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• pp.1-7
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• 2007

Medaka(Oryzias latipes) and earthworm(Eisenia fetida) toxicity tests were carried out and ecological risk assessment in water and soil was performed with national monitoring data. NOEC of alachlor was $100\;{\mu}g\;L^{-1}$ in the medaka early life-stage test. Embryonic development, hatchability and time to hatching of medaka eggs were affected by this chemical. The $LC_{50}$ and NOEC of alachlor were $94.1\;mg\;kg^{-1}\;and\;55.0\;mg\;kg^{-1}$, respectively, in the earthworm acute toxicity test. The environmental monitoring has been carrying out by NIER since 1999. Exposure levels of alachlor in water and soil were $ND{\sim}0.54\;{\mu}g\;L^{-1}\;and\;ND{\sim}0.9\;{\mu}g\;kg^{-1}$, respectively, in national monitoring data which had been performed from 2000 to 2004. The measured water and soil exposure levels were applied to evaluate the environmental risk assessment. The PNEC of alachlor in water and in soil were determined as $1\;{\mu}g\;L^{-1}\;and\;55.0\;{\mu}g\;kg^{-1}$, respectively using the safety factors which were suggested in EU and OECD. The HQs (PEC/PNEC) were determined to be below 1 for both water and soil when the maximum exposure levels ($0.54\;{\mu}g\; L^{-1}$ in water and $0.9\;{\mu}g\;kg^{-1}$ in soil) were applied. Conclusively, our study indicated that there was not significant ecological risk of alachlor in water and soil of our monitoring sites.

• Toxicological Research
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• v.14 no.3
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• pp.379-384
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• 1998

Endocrine disruptor is an exogenous substance that changes endocrine function and causes adverse effects at the level of the organism, its progeny, and/or (sub)populations of the organisms. Purported adverse effects are cancers, declines in reproductive health, developmental learning disabilities in humans, and declining populations, altered morphology, physiology or behavior in wildlife. In these days, expert groups on chemicals in IPCS, IFCS and OECD are intensively discussing the identification of endocrine disruptors and the proper management of those chemicals. In this study, we screened the endocrine disrupting effects of lead using fertilized eggs of Oryzias latipes. In brief, the eggs were exposed to lead with different concentrations at Ringer's solution, and the mortality, the incidence of deformation, the body movement and the hatching success were determined after incubation. The histological analysis of normal and deformed larvae was also carried out. Compared to control, the mortality and the heart rate of eggs and/or larvae increased, but the hatching success and the tail movement decreased. The morphological observation showed the asymmetrical deformation of larvae and the distortion of spinal cord. The absorption of the liquid in yolk sac was hindered. The adverse effects of lead in the ontogeny of fertilized eggs of Oryzias latipes seemed to be stronger in pH5.6 than in pH7.5 solution. In summary, lead showed adverse effects on the ontogeny of fish fertilized eggs plays critical role in regulating biological systems and controlling developmental processes as an endocrine disruptor.

• 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.

• Environmental Analysis Health and Toxicology
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• v.15 no.4
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• pp.131-137
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• 2000

The release of hazardous waste materials into the environment poses serious risks in humans and ecosystems. The risk assessment of environmental pollutants including hazardous chemicals requires a comprehensive measurement of hazard and exposure of the chemicals that can be achieved by toxicity evaluation using a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed . In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 bp encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 Up of mRNA was consistently amplified in all the treated samples including control There were no significant differences in the TH expression level regardless of treating concentrations (1∼5,000 ppb) and time (0∼48 hr) The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue - specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard ofvarious environmental pollutants.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2002.10a
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• pp.164-164
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• 2002

에스트로젠을 처리한 송사리의 간으로부터 choriogenin vitellogenin estrogen receptor의 발현량을 전사수준에서 Real-time을 사용하여 정량.비교하였다. 시험어종으로는 부화 후 5개월 이상된 성숙한 수컷 송사리(Oryzias latipes)를(체중 약 250mg/마리)를 사용하여 17$\beta$-estradiol(25ppt, 50ppt, 100ppt)에 24시간 노출시켰다. Fluorescence dye는 choriogenin vitellogenin estrogen receptor의 경우 FAM (6-carboxyfluorescein)을 사용하였으며, $\beta$-actin의 경우는 VIC를 사용하였다. 프로브에 사용하는 quencher dye는 TAMRA(6-carboxy-N',N',N',N'-tetramethyl rhodamine)을 사용하였다. Internal control로 사용된 $\beta$-actin은 17$\beta$-estradiol의 농도에 상관 없이 0~10pM 범위에서 일정하게 발현됨을 보여주었다. vitellogenin choriogenin L 및 choriogenin H는 17$\beta$-estradiol의 농도에 의존하여 발현이 증가되는 용량-반응양상(Dose-dependent)을 나타내었다. 반면, estrogen receptor는 모든 처리군에서 $10^{-2}$pM 정도로 발혐됨에 따라 본 시험농도의 17$\beta$-estradiol에 의해서는 거의 유도발현이 되지 않음을 보여주었다. choriogenin L, choriogenin H, vitellogenin I 및 estrogen receptor 발현감수성을 비교한 결과, 25ppt 및 50ppt의 17$\beta$-esoadiol 농도에서는 ChgL > ChgH > VTG I >ER의 순으로 감수성이 높았으며, 100ppt 노출에서는 ChgL > VTG I > Chg H > ER의 순으로 감수성이 높게 나타났다. 결론적으로 choriogenin이 에스트로젠물질에 의한 가장 민감한 생체지표임을 알 수 있었다.

• Environmental Analysis Health and Toxicology
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• v.20 no.1
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• pp.13-21
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• 2005

The purpose of this study was to determine the effects of bisphenol A (BPA), which is known to have estrogenic activity, on the early development of medaka fish (Oryzias latipes). The fertilized eggs of medaka were treated with BPA at different concentrations for 3 weeks. Embryonic growth, deformation, hatching success, and gonadal differentiation were determined to observe the effects of this chemical. Also we tried to measure the estrogenic activity of bisphenol A using ELISA and RT-PCR methods. By using this techniques, we evaluated the induction of vitellogenin, an estrogen-regulated gene from the whole body-homogenates of larvae. At results, a reduced blood circulation was seen in embryos and peritoneal edema and hindrance of yolk-sac absorption were observed in larvae of treated group. However, BPA at the concentrations tested (2～200 ㎍/L) did not have severe adverse effects on the early life-stages. According to the observation of gonadal histology, inter-sex or sex -reversal was not found in all test fish. After the exposure was ended, vitellogenin mRNA and protein levels were measured in larvae and then their levels were found to be increased in treated group with 200㎍/L. These results indicate that BPA can induce the expression of vitellogenin in early life-stages as well as in adult male fish.

• Korean Journal of Environmental Agriculture
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• v.7 no.2
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• pp.124-129
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• 1988

Prolonged aquatic toxicity tests (7-days) of six pesticides to the freshwater fish Oryzias latipes were performed to confirm the adequacy of the exposure times, 48-hr or 96-hr, which has been required by the protocols for the aquatic acute toxicity test. The toxicity curves were plotted for each chemical, and the significance of the difference between lethal threshold concentrations and 48-hr or 96-hr LC50's was analysed statistically. The lethal threshold concentrations of butachlor, fenobucarb, and chlorothalonil were clearly defined on the 5th day, 2nd day, and 2nd day at 0.53mg/1, 10.3mg/l, and 0.085mg/1, respectively. But the toxicity curves of alachlor, diazinon, and iprobenfos continued with no threshold for 7 days. Four out of six test pesticides failed to show the threshold concentration during the 96-hr exposure time. Therefore, the results of the acute toxicity test using 48-hr or 96-hr exposure time would not describe the ideal toxicity of pesticides. It is recommended that the prolonged exposure should be continued long enough to define the threshold adequately.

• The Korean Journal of Pesticide Science
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• v.15 no.4
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• pp.350-356
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• 2011

Environment-friendly agro-materials tend to be preferred to chemical insecticides recently. For this reason, many studies were conducted to develop environment-friendly insecticides containing natural materials. The purpose of this study was to assess ecotoxicity for 5 plant essential oils (Thyme white, Clove bud, Cassia, Lavender, Lemon eucalyptus) expected to prevent from pests and be used for agro-materials. Target species used to assess acute toxicity were aquatic invertebrate (Daphina magna), fish (Oryzias latipes), honeybee (Apis mellifera L.) and earthworm (Eisenia fetida). The EC50 value, toxicological responses of thyme white, clove bud, and cassia to Daphina magna were 2.5, 2.8, and $6.9mg\;L^{-1}$ respectively and these values were moderately toxic according to standard of USEPA. $EC_{50}$ of Lavender and lemon eucalyptus were >$10mg\;L^{-1}$ then they were considered as slightly toxicity. In case of acute toxicity test to fish, $LC_{50}$ of thyme white and cassia were 6.7 and $7.5mg\;L^{-1}$ each other. The other plant essential oils indicated $LC_{50}$ >$10mg\;L^{-1}$. Acute contact and oral toxicity test to Honeybee were conducted. As a result, $LD_{50}$ of all essential oils were >$100{\mu}g$ a.i. $bee^{-1}$ in both of tests. In case of acute toxicity test to earthworm, $LC_{50}$ of thyme white, clove bud, cassia, lavender, and lemon eucalyptus were 149, 230, 743, 234, and $635mg\;kg^{-1}$, respectively. In conclusion, if the safety for earthworm is confirmed, 5 plant essential oils are expected to be use for environment-friendly insecticide materials with low risk against ecosystem and contribute to developing environment-friendly agro-materials.