Evaluation of Aquatic Toxicity Induced by Environmental Contaminants Using High-sensitivity Method for Detecting DNA Damage

利用高灵敏度 DNA 损伤检测方法评价环境污染物引起的水生毒性

• 批准号: 09680532
• 负责人: IWAHORI Keisuke
• 金额: $ 1.86万
• 依托单位: University of Shizuoka
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680532/
• 关键词: comet assay; DNA damage; DNA repair; Euglena gracilis; sensitivity; hazardous chemical; dye exclusion test; lymphocyte of human; MNNG; BAP; 電気泳動; unwinding; r線照射; Tail length; Tail moment; Lysis; タイプカルチャー; リンパ球; γ線照射

This research aimed at the applicability of environmental microorganisms to comet assay and its availability for evaluation of aquatic toxicity induced by environmental contaminants. The obtained results can be summarized as follows ;Euglena gracilis, Chlamydomonas pulsatilla, Saccharomyces cerevisiae, Monas guttula and Tetraphymena pyriformis were targetted. From the results of cell lysis operation, 0.01% SDS solution was applicable to E. gracilis, 0.05% SDS solution was to C. pulsatilla and 1% Triton X-100 solution was to T. pyriformis, respectively. Changing with various times of cell lysis, unwinding and electrophoresis, applicabilities of those microorganisms to comet assay were investigated. Consequently, E. gracilis was found to be most suitable and its operational modes was determined that cell lysis time was 5 min. and each time for unwinding and electrophoresis was 20 min., respectively.Evaluations of DNA damage and repair to hazardous chemical substances were carried out with comet assay using E. gracilis. Within the range of each molarity at which the survival of E. gracilis could be confirmed by dye exclusion test, DNA damages were detectable for MNNG and BAP, but were not for MMC and AMD. Using MNNG-treatment and γ-irradiation, the results of DNA repair evaluation were shown that DNA could be repaired within 1 hr. Comparing with the results by lymphocyte of human, more than 10-fold and 2-fold sensitivities in MNNG- and BAP-treatments, respectively, were shown in comet assay using E. gracilis.Judging from the above-mentioned results, it was concluded that comet assay using E. gracilis, developed by this research, was applicable to evaluate aquatic toxicity induced by environmental contaminants.

本研究旨在探讨环境微生物对彗星试验的适用性及其在环境污染物对水生生物毒性评价中的应用价值。所获得的结果可以概括如下：Euglena gracilis，Chlamydium pulsatilla，Saccharomyces cerevisiae，Monas guttula和Tetraphymena pyriformis。从细胞裂解操作结果来看，0.01%SDS溶液适用于大肠杆菌。gracilis，0.05%SDS溶液对C.白头翁和1%Triton X-100溶液对T.梨形肌。考察了不同裂解时间、解旋时间和电泳时间对彗星试验的影响。因此，E.结果表明，Gracilis最适合于细胞裂解，其操作模式为裂解时间5 min，解旋电泳时间20 min，用彗星试验评价有害化学物质对DNA的损伤和修复作用。股薄肌在E.染料排斥试验可证实股薄肌损伤，MNNG和BAP可检测到DNA损伤，而MMC和AMD未检测到DNA损伤。用MNNG处理和γ射线照射的DNA修复评价结果表明，DNA可在1小时内修复，用大肠杆菌彗星试验比较，MNNG和BAP处理的敏感性分别为人淋巴细胞的10倍和2倍以上。从上述结果可以看出，用E。本研究开发的gracilis适用于评价环境污染物对水生生物的毒性。

项目成果

岩堀恵祐: "EICA年報Vol.4(環境計測としてのバイオモニタリングの意義)"環境システム計測制御学会(予定). (2000)

Keisuke Iwahori：“EICA 年度报告第 4 卷（生物监测作为环境测量的重要性）”环境系统仪器和控制协会（计划）（2000 年）。

岩堀恵祐: "環境微生物を用いたコメットアッセイ操作条件の実験的検討"日本水処理生物学誌. 35巻. 261-270 (1999)

Keisuke Iwahori：“使用环境微生物进行彗星测定操作条件的实验研究”，《日本水处理生物学杂志》，第 35 卷，261-270（1999 年）。

Keisuke IWAHORI: "Significance of Bio-monitoring as Environmental Measurement"The Society of Environmental Instrumentation Control and Automation. (in press). (2000)

Keisuke IWAHORI：“生物监测作为环境测量的意义”环境仪器控制与自动化学会。

岩堀恵祐、宮田直幸、下位香代子、木苗直秀 他: "環境微生物を用いたコメットアッセイ操作条件の実験的検討"日本水処理生物学会誌. 35巻4号. 261-270 (1999)

Keisuke Iwahori、Naoyuki Miyata、Kayoko Shimo、Naohide Kinae 等：“使用环境微生物进行彗星测定操作条件的实验研究”日本水处理生物学学会杂志，第 35 卷，第 4. 261-270 期（1999 年）。 ）

岩堀恵祐: "EICA年報Vol.4(環境計測としてのバイオモニタリングの意義)"環境システム計測制御学会(印刷中). (2000)

Keisuke Iwahori：“EICA 年度报告第 4 卷（生物监测作为环境测量的重要性）”环境系统仪器和控制协会（印刷中）。