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应用 RAPD 技术检测呋喃唑酮对海洋纤毛虫(原生动物,纤毛门)车轮虫引起的 DNA 损伤。

Use of RAPD to detect DNA damage induced by nitrofurazone in marine ciliate, Euplotes vannus (Protozoa, Ciliophora).

机构信息

Laboratory of Protozoology, Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou 510631, Guangdong, China.

出版信息

Aquat Toxicol. 2011 Jun;103(3-4):225-32. doi: 10.1016/j.aquatox.2011.03.002. Epub 2011 Mar 15.

Abstract

The random amplified polymorphic DNA (RAPD) assay was evaluated as a potential tool to detect the ecotoxicity induced by nitrofurazone in marine ciliate, Euplotes vannus. The data revealed a reduction in viability of the test ciliates with increasing nitrofurazone concentration in the range of 0-24 mgl(-1) and time of exposure from 24 to 96 h. The nitrofurazone treated ciliates were subjected to DNA damage analysis by RAPD assay. Among the 33 test RAPD primers used in this study, 11 primers with 60-70% GC content produced unique polymorphic band patterns. A total of 213 bands of 155-3317 bp in molecular size range were observed in the untreated cells. In comparison with the control ciliates, the nitrofurazone treated groups showed differences in RAPD profiles with respect to the band intensity, disappearance of bands and appearance of new bands of amplified DNA. The variation of RAPD profiles showed both the time- and concentration-dependent relationships. The data suggested significant genomic template instability, which corresponds well with the viability of the test ciliates. Thus the results demonstrated the potential of the RAPD assay for application as a powerful tool for detecting genotoxicity induced by fishy drugs in aquatic environment.

摘要

随机扩增多态性 DNA(RAPD)分析被评估为一种潜在的工具,用于检测呋喃唑酮在海洋纤毛虫,游仆虫中诱导的生态毒性。数据显示,随着呋喃唑酮浓度在 0-24 mgl(-1)范围内和暴露时间从 24 小时增加到 96 小时的增加,试验纤毛虫的活力降低。用 RAPD 分析对经硝基呋喃唑酮处理的纤毛虫进行 DNA 损伤分析。在本研究中使用的 33 个测试 RAPD 引物中,11 个具有 60-70%GC 含量的引物产生了独特的多态性带型。在未经处理的细胞中观察到大小在 155-3317bp 之间的 213 个带。与对照纤毛虫相比,硝基呋喃唑酮处理组在 RAPD 图谱上表现出条带强度、条带消失和新扩增 DNA 条带出现的差异。RAPD 图谱的变化表现出时间和浓度依赖性关系。这些数据表明基因组模板不稳定,这与试验纤毛虫的活力相符。因此,结果表明 RAPD 分析有潜力作为一种强大的工具,用于检测水产环境中鱼药引起的遗传毒性。

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