血吸虫病疫区野生鱼体中甲状腺激素干扰物的效应评价与种类鉴别

Large number of technical sodium pentachlorophenate have been sprayed around typical regions of schistosomiasis prevalence to kill Oncomelania hupensis Gredler. Many organic byproducts will be carried into local aquatic environment.These pollutants may induce thyroid hormones (THs) disrupting effects in wildfish.It is difficult to identify thyroid disrupting chemicals (TDCs) and primary contributors in wildfish in the absence of effective method. The biomarkers of thyroid disrupting effect in fish serum will be detected, the two-hybrid thyroid hormone receptor (TR) yeast assay was also used to quantify agonistic and antagonistic TR activities of these TDCs in fish muscle tissue. The combined disrupting effect caused by TDCs were studied by both in vivo and in vitro bioassays. Column chromatographys were applied to separate and purify typical fractions, each of these fractions eluted with different solvents from column chromatographys were bioassayed by the two-hybrid TR yeast, these fraction will be analyzed by instruments which the disrupting effect were detected in; the correlations between effect and TDCs levels were explored by statistical method; the primary contributors will be identified. The item will develop a method for assessment the combined thyroid disrupting effect and identification of chemicals responsible for it in wildfish from typical regions of schistosomiasis prevalence. The results will be beneficial to control and minify these objective TDCs in the study regions, and to develop China guidance method of assessment the combined thyroid disrupting effect and identification of chemicals responsible for it.

血吸虫病疫区因大规模使用工业五氯酚钠灭螺，导致五氯酚钠生产过程中伴生的有机副产物也被大量喷洒进了水环境中。多种污染物的复合暴露可能诱导疫区野生鱼体产生甲状腺激素(HTs)干扰效应，但鱼体内富集的有机污染物众多，由于缺少解析化学污染与干扰效应之间关系的方法，难以鉴别引起HTs干扰效应的化合物种类及主要效应因子。本申请拟基于离体/活体两种生物毒性测试技术，评价鱼体中的THs干扰效应；通过优化多种填充柱分离纯化技术和仪器分析技术，鉴定鱼肉组织复杂的有机萃取物中甲状腺激素干扰物(TDCs)的种类；并探索干扰效应和污染物之间的定量关系，识别主要效应因子；归纳出一套鱼体中THs干扰效应评价与干扰物种类鉴别的方法。为我国TDCs毒性鉴定和效应评价体系规范的建立与完善提供前沿性的研究基础，为疫区典型TDCs的控制与消减提供重要的参考数据，具有重要的理论创新和现实意义。

血吸虫病疫区因大规模使用工业五氯酚钠灭螺，导致五氯酚钠生产过程中伴生的有机副产物也被大量喷洒进水环境将危害水生生物的安全。本项目通过建立典型甲状腺激素干扰物（PCBs, CBs, CPs, PBDEs和HBCDs）的定性定量分析方法,用仪器定量分析洞庭湖疫区和非疫区对照点的鱼类样品，识别疫区野生鱼体中诱导产生甲状腺激素干扰效应的主要因子。.主要成果如下: (1)优化了GPC分离PCBs, CBs, CPs, HBCDs和PBDEs的时间。PCBs, CBs, CPs, HBCDs和PBDEs在GPC上洗脱液最佳收集时间分别为6-13 min, 8-15 min, 4-11 min, 6-13 min和8-14 min，因此同时收集5类化合物洗脱液的最佳时间为4-15 min。.(2)优化了PCBs, CBs, CPs, HBCDs和PBDEs前处理与分析技术。PCBs, CBs和PBDEs的样品净化选用10%AgNO3硅胶,3.3%去活化硅胶和44%酸化硅胶; HBCDs和CPs的样品净化选用44%酸化硅胶。通过优化色谱的分离条件和加标回收率实验验证建立了GC-MS/MS定量分析PCBs,CBs和CPs技术; GC-MS (ENCI)分析PBDEs技术，LC-MS/MS分析HBCDs技术。.(3)根据仪器分析结果识别了甲状腺激素干扰效应的主要因子。运用建立的GC-MS/MS和LC-MS/MS分析技术,定量分析具有代表性的血吸虫病疫区（湖南省益阳市茅草街镇）和非疫区对照点（桃花江水库）的鱼样共130尾。前处理过程中将鱼样分为46个，其中疫区鱼样25个；非疫区鱼样21个。疫区鱼样中污染物的含量依次为∑CBs> ∑CPs > ∑PCBs > ∑PBDEs > ∑HBCD, 疫区鱼样中∑CBs的含量显著高于非疫区对照点的含量, 结合5类化合物的甲状腺激素干扰效应, 总结出CBs是鱼样体内的主要效应因子。运用建立的TDCs种类鉴别方法检测出的数据将为疫区典型TDCs的控制与消减提供重要的参考数据。