应用重组孕激素基因酵母测定饮用水中内分泌干扰物的方法

研究了用重组孕激素受体基因酵母测定饮用水中内分泌干扰物的方法,并利用该种方法检测了南方某水厂不同处理工艺过程水样对孕激素受体活性的抑制水平.结果表明,重组孕激素受体基因酵母能够与孕激素专一性的结合,诱导产生明显的剂量-效应关系,EC₅₀值为0.5 nmol/L,具有较高灵敏度;环境内分泌干扰物五氯酚和壬基酚具有孕激素受体抑制活性,其IC₅₀值分别为2.4μmol/L和3.7μmol/L;重组孕激素受体基因/报道基因的酵母技术是一种筛选和定量分析具有孕激素受体抑制活性的内分泌干扰物的快速、有效方法.结合固相萃取的前处理技术,重组孕激素受体基因酵母对水厂不同处理工艺水样检测出明显的孕激素受体抑制活性,抑制率均大于58%,表明重组孕激素受体基因酵母检测技术能够快速监测和鉴别水样中具有抑制孕激素受体活性的物质.     

The Precautionary Principle and Chemicals Regulation: Past Achievements and Future Possibilities (8 pages)

Background, Aim and Scope

The paper investigates the development of the institutional basis of the present modes of chemicals regulation and management, with special attention to interrelations with the precautionary principle. Main Features: The paper elucidates on how the precautionary principle has been shaped in relation to chemicals regulation and management since Carson's Silent Spring (years before the principle was confirmed as a policy-principle in German and European legislation. Furthermore, it is examined how the precautionary principle interacted with the development of the present chemicals regulatory regime, in a complex interplay within the OECD and Member Countries. The present modes of precaution in the new EU chemical legislation – REACH – are investigated with respect to the precautionary principle, and tested against two contemporary problems; brominated flame retardants and endocrine disrupting substances. Results: The analysis demonstrates the changing character of the integration of the precautionary principle. The main tendencies are from implicit to more explicit precaution and from a closed expert-orientation towards a more deliberative approach to scientific knowledge and uncertainty. The results demonstrate that the precautionary principle is manifest in both the design of the testing strategy and in policy provisions. In particular, the substitution of hazardous substances with less hazardous is important. Discussion: Despite explicit attention to the precautionary principle, is the present reformulation of the European Chemicals policy in danger of falling into loop-holes that equal problems related to the present regulation of existing chemicals? 'Precaution' has been reduced virtually to an abstract concept that is more or less devoid of practical meaning in the regulatory process. Conclusions: It is concluded that the role of the precautionary principle in chemicals regulation will require continued scrutiny in the future shaping of the REACH strategy. Perspectives: Continued development of robust and precaution-based chemicals regulation will have to involve both new data-generation strategies and new forms of political decision-making, with special attention given to transparency and deliberative policymaking.     

孕激素左炔诺孕酮长期低剂量暴露对雄性斑马鱼的生殖毒性

孕激素是广泛使用的高生物活性药物,其残留广泛存在于水环境中,对水生生物造成潜在威胁。本论文研究孕激素左炔诺孕酮(LNG)长期暴露对雄性斑马鱼的生殖毒性。将斑马鱼胚胎暴露于环境相关浓度的LNG(10, 33, 100 ng·L~(-1))至性成熟(受精后142 d),得到100%的雄性斑马鱼。将得到的雄鱼与未暴露的雌鱼配对,结果发现100 ng·L~(-1) LNG处理组雄鱼使得未暴露雌鱼产卵的数量较对照组雄鱼使得未暴露雌鱼产卵的数量显著减少。33和100 ng·L~(-1)处理组雄鱼与未暴露雌鱼配对所得子代在受精后7 d内的存活率较对照组雄鱼与未暴露雌鱼配对所得子代的存活率显著降低。33 ng·L~(-1) LNG显著升高总活力、活跃精子活力、平均路径速度、直线运动速度、曲线运动速度等精子活力参数,但其他浓度的LNG并不影响精子的活力。LNG在33和100 ng·L~(-1)的浓度下显著降低雄鱼血浆中11-酮基睾酮(11-KT)的含量,且在3个浓度下均显著降低雄鱼血浆中雌二醇(E2)的含量,但对睾酮(T)无明显影响。此外,LNG暴露可干扰雄鱼大脑和精巢中孕激素受体(npr, mprα, mprβ)、雌激素受体(erα, erβ)以及雄激素受体(ar)的转录,可能通过这些受体途径参与生殖调控。研究结果表明环境相关浓度的左炔诺孕酮长期暴露可导致雄性斑马鱼的生殖内分泌干扰效应,并提示:仅雄鱼单独受孕激素暴露影响,也可能对鱼类种群的生殖产生深远影响。     

除草剂乙草胺对非洲爪蟾性腺发育的影响

使用性腺发育对内分泌干扰作用敏感的两栖动物非洲爪蟾作为模型动物,通过考察性别比、性腺整体形态和组织学形态来揭示乙草胺对其性腺发育的影响,从而确定乙草胺是否具有内分泌干扰活性.46/47期蝌蚪经不同浓度(5μg·L-1、10μg·L-1、20μg·L-1)的乙草胺暴露处理至变态后1个月,在变态结束后3个月时,解剖性腺、鉴别雌雄并固定性腺做组织切片.实验中对照组所有性腺都为典型的卵巢或睾丸,雌性百分率为48.78%(20/41).而乙草胺暴露组却出现了明显的异常睾丸,其总体上是睾丸,但又带有部分卵巢的特征,在统计性别时将具有异常睾丸的爪蟾归为雄性.5μg·L-1、10μg·L-1、20μg·L-1暴露组的雌性百分率分别为:57.14%(24/42)、66.67%(22/36)、55.26%(21/38).虽经统计分析无显著性差异,但乙草胺暴露组中雌性多于雄性的趋势和出现带有雌性特征的雄性的现象,一定程度上却暗示了乙草胺可能对非洲爪蟾的性腺发育有雌性化作用.正常睾丸的组织学结构应有发育成熟的生精小管,其内有各个时期的精子囊和生精细胞及精子.但经乙草胺处理的爪蟾睾丸却表现出几种明显的异常:没有生精小管,相反却像发育早期的睾丸一样,充满大量的精原细胞,完全没有生精细胞和精子;有生精小管结构,但其内精子囊结构不明显,生精细胞少并且排列混乱,其间有很大的空隙;睾丸出现腔隙,并有大量体细胞存在,这种结构与57期的卵巢相似;睾丸中出现异常的生精细胞,与58期卵巢的卵细胞相似;睾丸组织中有明显的卵细胞散布.组织学研究发现的乙草胺导致的睾丸结构证明了乙草胺对非洲爪蟾的睾丸发育有抑制雄性化作用和雌性化作用,即乙草胺对非洲爪蟾的性腺发育有内分泌干扰作用.     

手性污染物与环境安全

本文就手性污染物在生命过程中对映体的差异作了简要评述,分析了手性污染物对映体的毒性差异及机理.同时指出了手性污染物今后研究的发展趋势.     

Sorption of the natural endocrine disruptors,oestrone and 17 beta-oestradiol in the aquatic environment

Adsorption of hydrophobic contaminants at the particle/water interface is one of the key processes controlling their fate in the aquatic environment. The sorption of the natural female hormones oestrone and 17-oestradiol has been studied under simulated riverine conditions. Both the kinetics and the effects of varying fundamental environmental parameters (e.g. sediment properties) on the thermodynamic equilibrium partition coefficient (K _p) have been studied in continuous and batch sorption experiments, respectively. Results showed that the sorption of oestrone and 17-oestradiol by sediment was relatively slow, reaching equilibrium in 50 days. In addition, relatively small adsorption of both oestrone and 17-oestradiol onto the sediment was observed, with K _p values between 200 and 250 mL g^–1. The comparable K _p values of the two compounds reflect their structural similarity. It can be concluded that the two endocrine disruptors, oestrone and 17-oestradiol remain primarily in association with the aqueous phase.     

Metabolism of a nonylphenol isomer by Sphingomonas sp. strain TTNP3

For elucidation of the metabolism of the endocrine disruptor nonylphenol by Sphingomonas sp. strain TTNP3, the degradation of an isomer of nonylphenol, 4(2,6-dimethyl-2-heptyl)-phenol, has been studied. As in the case of 4(3,5-dimethyl-3-heptyl)-phenol, the metabolism of this nonylphenol isomer leads to the formation of the NIH-shifted product 2(2,6-dimethyl-2-heptyl)-1,4-benzenediol (NIH: National Institute of Health), but also to the alkoxy derivative 4(2,6-dimethylheptan-2-yloxy)phenol as additional metabolite. To the best of our knowledge, this is the first report describing the formation of alkoxyphenol as a degradation product of nonylphenol. Additionally, these results provide for the first time evidence for slight differences in the biodegradation of the isomers of nonylphenol.     

Endocrine disruption of water and sediment extracts in a non-radioactive dot blot/RNAse protection-assay using isolated hepatocytes of rainbow trout

Goal, Scope and Background In order to evaluate the estrogenic activity of sediments and XAD water extracts of selected sites of the catchment area of the River Neckar, a river system in Southern Germany, an integrative assessment approach was used to assess the ecological hazard potential of endocrine-disrupting compounds in sediment and water. Methods The approach is based on estrogen receptor-mediated vitellogenin synthesis induced in isolated hepatocytes of rainbow trout and quantified in a non-radioactive dot blot/RNAse protection-assay in parallel to comprehensive chemical analyses of estrogenic substances. Results and Discussion Numerous investigated extracts revealed an estrogen activity comparable to that of the positive control (1 nM 17?-estradiol corresponding to 270 ng/L in the test medium). Based on a concentration factor of 30 in the extracts and a recovery of XAD resins of approximately 80 %, 17?-estradiol equivalent concentrations between 20 and 26.7 ng/L could be calculated downstream of a sewage treatment plant (< 0.1 ng/L for a reference site). A comparison of the bioassay-derived Bio-TEQs (toxicity equivalents) and the Chem-TEQs revealed a high correlation with a Pearson coefficient of 0.85, indicating that the same ranking of the samples could be obtained with respect to the endocrine disrupting potential with both chemical and bioanalytical analysis. However, the TEQ concentrations computed from chemical analyses were significantly lower than the bioassay-derived TEQ concentrations. In fact, in none of the samples, more than 14 % of the vitellogenin-inducing potency could be attributed to the substances (steroids, alkylphenols, bisphenol A, diethylstilbestrol) analyzed. A comparison of the endocrine disrupting potential of sediments extracted by the solvents acetone and methanol revealed lower biological effects for acetone-extracted samples. Possible reasons may be a masking of endocrine effects in acetone extracts by cytotoxicity, a low extraction efficiency of the solvent acetone, or anti-estrogen potencies of some extracted sediment compounds. Using a mass balance approach, the contribution of the compounds analyzed chemically (Chem-TEQs) to the total endocrine activity (Bio-TEQs) was calculated. Based on the very low detection limits, particularly of the steroids with their high TEF factors, results revealed that a calculation of the Chem-TEQs is associated with considerable scale inaccuracy: Whereas only 7-15 % of the biological effectiveness (Bio-TEQs) could be explained by endocrine substances identified above the detection limits, the assumption of concentrations slightly below the given detection limits would result in a significant over estimation (137-197 %) of the Bio-TEQs. Even the interassay variation of the dot blot assay with different fish donors for primary hepatocyte (factor 2 - 2.5) is relatively low, when compared to the large range of the Chem-TEQ concentrations (factor 20) obtained when applying different modes of calculation. Conclusions and Outlook Overall, only a minor portion of the endocrine activity detected by bioassays could be linked to compounds identified by chemical analysis. In vitro assays for assessment of endocrine activities are useful as sensitive integrating methods that provide quantitative estimates of the total activity of particular receptor-mediated responses. Although discrepancies may also result from different bioanalytical approaches, it is overall likely that bioanalytical and not chemical analytical approaches give the correct estimate of endocrine disrupting potencies in environmental samples. As a conclusion, assessment of endocrine disruption based on chemical analysis alone does not appear sufficient and further research into the spectrum of substances with potential endocrine activity as well as into additive or even synergistic effects in complex environmental samples is urgently needed.     

改性无烟煤去除水中内分泌干扰物的性能探究

以KOH为改性剂对无烟煤改性,研究了改性前后无烟煤对3种内分泌干扰物EDCs(17α-乙炔基雌二醇EE2、双酚A BPA、磺胺甲噁唑SMZ)的吸附性能。结果表明,KOH改性对无烟煤的孔结构和表面性质产生影响,提高了无烟煤对3种EDCs的吸附量,其中对EE2的吸附量提高了近2倍。3种EDCs在改性前后2种无烟煤上的吸附过程均可用伪二级动力学描述,吸附等温线更符合Langmuir吸附等温方程,表现为化学吸附。改性前后2种无烟煤对EE2均表现出最高吸附能力,平衡吸附量(qe)分别为0.214 7μg/mg与0.622 9μg/mg;其次是BPA;最差是SMZ,qe仅为0.095 9μg/mg与0.124 2μg/mg。进一步研究表明,目标物的吸附情况与其憎水性质和分子结构有关,无烟煤吸附目标物的能力与其正辛醇/水分配系数正相关。     

杀菌剂三唑酮毒性及内分泌干扰作用研究进展

在对近年来国内外三唑酮研究的热点问题进行分析讨论的基础上,概述了三唑酮的检测、毒性、内分泌干扰作用等方面研究的进展工作。