饮用水处理过程遗传毒性评价方法研究与应用
文献类型：学位论文
| 作者 | 刘楠楠
|
| 学位类别 | 博士 |
| 答辩日期 | 2013-11 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 马梅，王子健 |
| 关键词 | 饮用水处理过程 遗传毒性效应 水源水 鱼外周血红细胞微核试验 drinking water treatments genotoxic effect source water piscine peripheral micronucleus test |
| 其他题名 | Development and application of evaluation methods for genotoxicity assessment in drinking water treatments |
| 学位专业 | 环境工程 |
| 中文摘要 | 水源水污染日益严重，饮用水深度处理工艺应用越来越广泛，但臭氧氧化等替代消毒工艺过程中可能会产生新型污染物，特别是具有遗传毒性的污染物越来越引起关注。遗传毒性效应涉及多通路、多层次，作用方式具有复杂性和复合性，而目前饮用水遗传毒性评价指标单一，不能全面分析毒性潜力，并且可能存在假阴性结果。对饮用水处理过程开展多终点遗传毒性评估，研究饮用水存在潜在的安全风险十分重要。 本文针对饮用水遗传毒性效应检测，优化了表征染色体/纺锤体损伤的基于流式细胞仪体外微核自动化测试和建立了活体暴露稀有鮈鲫外周血红细胞微核检测技术，同时对表征 DNA损伤/修复的 SOS/umu实验进行优化。利用所建立的方法对我国北方、南方和中原地区 4个城市中 5个水源地和 9个饮用水厂处理过程及部分管网水在 DNA水平和染色体水平的遗传毒性特征进行分析，结合损伤作用类型分析遗传毒性效应的成因，主要结论如下： 将所优化的基于流式细胞仪体外微核自动化测试技术应用于饮用水中典型遗传毒物单一化学品检测，对中国仓鼠卵巢细胞（CHO k1）的染色体损伤效应评价发现碘乙酸 >溴乙酸>三溴乙酸>>二氯乙酸；对于 SOS/umu实验表征的DNA损伤/修复效应证实存在同样规律。将所建立的中国特有实验生物稀有鮈鲫外周血红细胞微核试验，同青鳉等其他模式鱼类对比，发现稀有鮈鲫对饮用水中遗传毒性物质具有更高敏感度，且适用于饮用水染色体损伤评价，为后续在线生物监测饮用水处理过程遗传毒性效应提供技术支撑。 对 5个典型水源地进行遗传毒性效应研究发现，五种水源水直接和间接DNA损伤/修复效应的 TEQ4-NQO和 TEQB[a]P范围分别是 0.02-0.18μg/L和0.10-0.46μg/L；水源水中非整倍体毒剂和染色体断裂剂共同造成染色体损伤，不同水源地两类物质的贡献存在差异。全二维气相色谱飞行时间质谱分析 3个水源地发现 DNA水平和染色体水平遗传毒性效应均最强的样点污染物种类和遗传毒性物质明显高于其他样点。 对 4个城市 9个饮用水厂处理过程遗传毒性特征研究发现，饮用水消毒工艺出水不同遗传毒性终点结果差异明显；氯化消毒出水 DNA损伤/修复效应显著，而臭氧氧化出水可能会增强染色体损伤效应，其主要贡献物质为染色体断裂剂；管网水随传输距离的不同遗传毒性效应存在差异。综上表明饮用水遗传毒性评价需要多终点、多层次检测体系。 |
| 英文摘要 | Due to serious aquatic environmental pollution in source water, new techniques in reinforced and advanced treatments are developed. However, it is still lack sufficient evidence in evaluation of genotoxicity in drinking water when emerging pollutants formed in alternative disinfectantion, e.g. ozonation. Genotoxic effects refer to multiple pathways and levels, and their modes of action are complex and combined. Recent applied index in genotoxicity evaluation of drinking water was single or limited, which may lack the ability to comprehensive evaluate the genotoxic potential and lead to false-negative responses. It is important to develop multi-endpoints evaluation methods for genotoxicity assessment in drinking water treatments. This paper optimized in vitro micronucleus assay in mammalian cells based on flow cytometric scoring, and established in vivo micronucleus assay for peripheral erythrocytes in Chinese Rare Minnow (Gobiocypris rarus). And SOS/umu test was optimized. The genotoxicity of five source waters and nine drinking water plants in China was evaluated, which were both in DNA and chromosomal levels. Analyzing the cause of genotoxicity in combination with modes of action in the generation of genotoxicity, we got the following conclusions: The in vitro automated scoring micronucleus assay was applied in evaluating the genotoxicity of typical haloacetic acids in drinking water. The rank order of chromosomal damage in Chinese hamster ovary cells (CHO k1) was iodoacetic acid> bromoacetic acid> tribromoacetic acid>>dichloroacetic acid. And the result was further confirmed by the optimized SOS/umu test. In addition, Chinese rare minnow seems to be more sensitive than other fishes, such as Japanese Medaka,when it’s applied in evaluating the chromosomal damage potential of drinking water. The present study supports the suitability of piscine micronucleus assay using Chinese rare minnow for on-line monitoring the genotoxic effect in drinking water. The results of genotoxicity in 5 typical source waters showed that the values of TEQ4-NQO and TEQB[a]P were 0.02-0.18μg/L and 0.10-0.46μg/L, which presented the direct and indirect DNA damage/repair effects, respectively. Aneugens and clastogens existed in source water, causing chromosomal damage, and their contribution varied in differen samples. For source water sample with the highest genotoxicity at both DNA and chromosomal levels, GC×GC-TOFMS analysis was used and showed highest number of pollutants and genotoxic substances than the other two samples. Genotoxicity assessment of nine drinking water treatment plants in four cities found that different endpoints in effluents of disinfection process showed different results. DNA damage/repair effect was significantly high in chlorination, while the chromosomal damage may increase in ozonation process, which was mostly contributed by clastogens. The genotoxic effect may change in the distribution system along with distance. All the results suggested that the combination of multi-endpoints test was needed in the genotoxicity evaluation of drinking water. |
| 公开日期 | 2015-07-08 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/15722]  |
| 专题 | 生态环境研究中心_环境水质学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | 刘楠楠. 饮用水处理过程遗传毒性评价方法研究与应用[D]. 北京. 中国科学院研究生院. 2013. |
入库方式： OAI收割
来源：生态环境研究中心
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