Avishai Nanthawan, Rabinowitz Claudette, Moiseeva Elisabeth, Rinkevich Baruch
Minerva Center for Marine Invertebrate Immunology and Developmental Biology, National Institute of Oceanography, Israel Oceanographic and Limnological Research, Tel-Shikmona, P.O. Box 8030, Haifa 31080, Israel.
Mutat Res. 2002 Jun 27;518(1):21-37. doi: 10.1016/s1383-5718(02)00069-4.
The alkaline comet assay, a sensitive method for DNA strand breaks and alkali labile site detection in individual cells, was employed here as an ecotoxicological monitoring tool for evaluation of genotoxicity in the Kishon River, Israel. This river, the most polluted river in Israel, has recently elicited major public concern with regard to cancer incidences in people who have dived there over many years. Five water samples were collected every odd month throughout the year 2001, from four localities. The comet assay was employed on fish hepatoma cell line RTH-149. Cells were exposed for 2h, in triplicate, to Kishon water: medium (1:1) samples that were pre-adjusted for pH and salinity percentage levels. Three DNA damage parameters (comet percentages, score of damage, and cumulative tail lengths of the comet), revealed significantly higher genotoxic values in Kishon water-treated cells as compared with the controls (up to 2.4, 3.2, and 3.6-fold, respectively). Part of the sampling sites revealed higher genotoxicity than other polluted sites. The results of this study demonstrate that the comet assay with RTH-149 cells can be successfully applied to a variety of aquatic samples revealing freshwater, marine and estuary conditions. The method found to be fast, sensitive, and suitable for monitoring programs.
碱性彗星试验是一种用于检测单个细胞中DNA链断裂和碱不稳定位点的灵敏方法,在此被用作一种生态毒理学监测工具,以评估以色列基顺河的遗传毒性。这条河是以色列污染最严重的河流,近年来,它引发了公众对多年来在此潜水人群癌症发病率的重大关注。2001年全年每隔一个月从四个地点采集五个水样。对鱼类肝癌细胞系RTH - 149进行彗星试验。将细胞一式三份暴露于预先调节pH值和盐度百分比水平的基顺河水与培养基(1:1)的样本中2小时。三个DNA损伤参数(彗星百分比、损伤评分和彗星的累积尾长)显示,与对照组相比,经基顺河水处理的细胞中的遗传毒性值显著更高(分别高达2.4倍、3.2倍和3.6倍)。部分采样点显示出比其他污染点更高的遗传毒性。本研究结果表明,用RTH - 149细胞进行的彗星试验可成功应用于各种水生样本,揭示淡水、海洋和河口环境状况。该方法被发现快速、灵敏且适用于监测项目。