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임정택 ( Jeongtaek Im ),전경미 ( Kyongmi Chon ),백민경 ( Min Kyoung Paik ),박경훈 ( Kyung-hun Park ) 한국환경농학회 2016 한국환경농학회 학술대회집 Vol.2016 No.-
최근 농약사용이 꿀벌에 미치는 영향에 대한 연구가 활발해지고 있으며 OECD에서는 꿀벌 유충 독성시험법 및 semi-field에 대한 시험법 등 꿀벌 안전성을 확인하기 위한 다양한 시험법을 구축하고 있다. 국내에서도 농약등록단계의 꿀벌 유충 독성시험법 도입의 필요성 및 타당성을 검토 중에 있으며 시험조건 확립에 대한 연구도 같이 진행하여 유충 단일노출 독성시험법에 대한 연구결과가 발표되기도 하였다. 본 연구에서는 최근 OECD 가이드라인에서 개정된 꿀벌 유충 반복노출 독성시험법에 대해 구축하고자 하였다. 본 시험법의 음성대조군은 증류수와 acetone을 사용하였으며 양성대조물질로는 dimethoate과 fenoxycarb를 사용하였다. 식이에 포함된 dimethoate의 양은 0.054 ㎍/㎕ diet, fenoxycarb의 양은 0.36ng/㎕ diet가 되게 하였다. 각 시험은 48 well plate에 3봉군에서 가져온 유충 1령을 최소 12마리씩 3반복으로 36마리 이상이 되게 하여 이충하였다. 시험 첫날 (D1)에 well plate에 이충용기(grafting cell)를 넣고 유충먹이 A를 20㎕, 시험3일차(D3)에 유충먹이 B에 시험용액을 혼합하여 20㎕ 공급하였고 시험 4-6일차에는 유충먹이 C에 시험용액을 혼합하여 30, 40, 50㎕ 씩 공급하였다. 시험기간 동안 plate를 34~35℃ 상태의 항온인큐베이터에 설치된 밀폐형 desiccator cabinet에 넣어 시험하였다. 유충기간동안(D1-D8) K2SO4 포화용액을 사용하여 95%±5% relative humidity (RH)를 유지하였으며, 용단계(pupation, D8-D15)에서는 NaCl 포화용액을 이용하여 80%±5% RH로 우화단계(emergence, D15-D22)에서는 50% 상대습도를 유지하였다. 우화단계에서는 자당용액을 공급할 수 있는 우화박스에 plate를 옮겨 관찰하였다. D4-D8에서 유충치사율과 D22에서 우화율을 측정하여 음성대조군과 양성대조군이 시험법 기준에 적합한지 확인하였다. OECD guideline에 따르면 음성대조군의 유충 누적치사율은 15% 이하, 우화율은 70% 이상이 되어야 하며 양성대조군 dimethoate의 유충 누적치사율은 50% 이상, fenoxycarb의 우화율은 20% 이하가 되어야 한다.
Mancozeb와 Pendimethalin에 의한 CHL Cell의 소핵생성과 세포사멸 여부 규명
문지홍 ( Ji-hong Moon ),전경미 ( Kyongmi Chon ),백민경 ( Min-kyoung Paik ),박경훈 ( Kyung-hun Park ) 한국환경농학회 2018 한국환경농학회 학술대회집 Vol.2018 No.-
Mancozeb is a widely used broad-spectrum fungicide known to be genotoxic and induces tumors in rodents at various sites. Pendimethalin is a member of family dinitroanaline of herbicides. Pendimethalin are well known to be more or less toxic to all living organisms. In this study, we investigated whether mancozeb and pendimethalin induce micronucleus and apoptosis in CHL (Chinese hamster lung) cells through flow-cytometry analysis for high throughput. We identified mancozeb (20, 40, 80 ug/ml) and pendimethalin (5, 10, 20 ug/ml) mediated micronuclei formation and apoptosis in a dose dependent manner. We evaluated an increase of micronuclei formation and apoptosis by flow-cytometry analysis and microscopy analysis. We identified that MMC (mitomycin) induced micronuclei formation as positive control and negative control (DW) has no effect on CHL cells. We demonstrated that mancozeb and pendimethalin may induce micronuclei formation and apoptosis in CHL cells. We also suggest that flow-cytometry analysis could be efficient high-throughput method for quantifying the formation of micronuclei.
The Correlation Between Deltamethrin Exposure and Urinary 3-PBA Concentrations in Rats
김아름누리 ( Areumnuri Kim ),박지현 ( Ji Hyun Park ),전경미 ( Kyongmi Chon ),박경훈 ( Kyung-hun Park ),문병철 ( Byeong-chul Moon ),백민경 ( Min Kyoung Paik ) 한국환경농학회 2016 한국환경농학회 학술대회집 Vol.2016 No.-
Pyrethroids (PYRs) are a widely used insecticide in agriculture and household area.PYRs caninduce neurotoxic effects in mammalsby disrupting voltage-sensitive sodium channelswhen exposed to overdose.In mammals, PYRs such as deltamethrinis metabolized to 3-phenoxybenzoic acid (3-PBA) in liver that is mainly excreted in urine.Urine is highly useful as a testing matrix because it is easily accessible and can be collected noninvasively. Little studies have reported on the PYRs-dosedependent effects on urinary 3-PBA.This study is designed to identify the correlation of deltamethrinamount exposured orally and dermally to rats and its metabolite(3-PBA) excreted in urine.This study is designed to exposure deltamethrin to rats in a dose-dependent manner and identify the correlation between deltamethrin exposure and its metabolite(3-PBA) in urine. Exposure levels consisted of three concentration of deltamethrin; control (0mg/kgbw), low (0.0705 mg/kgbw), medium (0.705 mg/kgbw) and high(7.05 mg/kgbw) dose. Low concentration of deltamethrin was calculated on the basis of Korea predictive operator exposure model (KoPOEM), indicating the field-realistic exposure to pesticides for farmer when mixing and applying pesticides. Single exposure of deltamethrin to Spraque-Dawley rats were conducted through oral and dermal pathway. Dermal exposure persisted for 6 h, and urine specimens were collected for 24h after exposure of both two pathway. The following procedures including hydrolysis, extraction and derivatization were conducted prior to the analysis of urinary 3-PBA. After acidic hydrolysis for the cleavage of conjugates, the analytes3-PBAwas extracted from urine matrix using n-hexane. Then, 3-PBA was derivatized to volatile estersusing N-tert.-butyldimethylsilyl-N-methyltrifluoroacetamid (MTBSTFA). Separation and detection was carried out using GC/MS. 2-Phenoxybenzoic acid served as internal standard for the quantification of 3-PBA. The limit of quantificationfor all analyteswas 0.98ng/ml urine.In the low-, medium- and high-dose oral deltamethrin groups, the concentrations of urinary 3-PBA were 44±2, 104±37, 943±65 ng/ml urine, respectively, showing good correlation between oral deltamethrin exposure and urinary 3-PBA in rats. This result was similar with those for dermal administration of deltamethrin. Based on these results, therefore, 3-PBA that is a main metabolite of PYRs may be suitable for urinary biomarker of exposure to PYRs.This study can be used as an index that can be predictedto exposuredosage of deltamethrinin the human from detection of urinary 3-PBA excretion.