Effects of estrone on the early life stages and expression of vitellogenin and estrogen receptor genes of Japanese medaka (Oryzias latipes)

The fertilized eggs of Japanese medaka (Oryzias latipes) were exposed to estrone (E1) at 5–5000 ng L⁻¹ for 15 d, and the hatched fry were exposed continuously to the same concentrations for the additional 15 d. Adverse effects on hatchability, time to hatching, and gross abnormalities occurred at 50 ng L⁻¹ or above. Then the fry were divided into a continual exposure group, and a water recovery group. When the fry were exposed to E1 for another 60 d, there was a decrease in the hepatosomatic index (HSI) of males and the influence disappeared in the water recovery group. The gonadosonatic index (GSI) of females at 500 ng L⁻¹ decreased significantly in another 60 d exposure. While the fry were maintained in dechlorinated tap water for 60 d, a significant decrease in female GSI was observed at 50 ng L⁻¹ or above. An increased GSI was found in males in both continual exposure and water recovery groups at all E1 treatments. Quantitative RT-PCR showed that vitellogenin-I (Vtg-I) gene expressions in the female liver were significantly down-regulated at 50 ng L⁻¹ in the continual exposure group, and at 500 ng L⁻¹ in the water recovery group, while male Vtg-I genes were significantly up-regulated for all E1 treatments. In addition, all E1 treatments caused sex reversal of males. These results suggest that E1 at 5 ng L⁻¹ or above have unrecoverable impacts on the gonadal growth and development of medaka, even if only early life stages were exposed to E1.     

Endocrine modulation and toxic effects of two commonly used UV screens on the aquatic invertebrates Potamopyrgus antipodarum and Lumbriculus variegatus

The two UV screens 3-benzylidene-camphor (3-BC) and 3-(4'-methylbenzylidene)-camphor (4-MBC) were tested regarding their toxicity and estrogenic activity. The Yeast Estrogen Screen (YES) and two sediment assays with the freshwater invertebrates Lumbriculus variegatus and Potamopyrgus antipodarum were performed. In the YES, both substances activated the human estrogen receptor alpha with EC50 values of 44.2 microM for 3-BC and 44.3 microM for 4-MBC, whereby 4-MBC attained only 8% of the maximal response of 17beta-estradiol. For P. antipodarum embryo production increased after exposure to both substances (EC50 of 4.60 microM 4-MBC=1.17 mg kg(-1)dw) while mortality increased at high concentrations. The reproduction of L. variegatus was decreased by 3-BC with an EC50 of 5.95 microM (=1.43 mg kg(-1)dw) and also by 4-MBC, where no EC50 could be calculated. While reproduction decreased, the worms' weight increased after exposure to 3-BC with an EC50 of 26.9 microM (=6.46 mg kg(-1) dw), hence the total biomass remained unaffected.     

Integral assessment of estrogenic potentials of sediment-associated samples

GOAL, SCOPE AND BACKGROUND: Exogenic endocrine-active substances are also called 'Endocrine Disrupting Chemicals' (EDC). They imitate or hinder the function of natural endogenic hormones or disturb the synthesis or the metabolism of hormones or of hormone receptors. The Enzyme-Linked Receptor Assay (ELRA) can detect estrogenic and anti-estrogenic effects at the level of receptor binding and is a useful tool for the integrative detection of contaminant effects. Although the test system has been used repeatedly in sediment assessments, the questions have remained concerning how it responds to variations in the physico-chemical matrix. For some bioassays, the salinity of the sample is a critical factor. This is especially relevant when testing wastewater samples or when sediment-associated samples in the tidal reaches of rivers are tested. Sediments in the tidal reaches of rivers change their salinity several times a day. Against this background, it would be beneficial to have a test procedure of known salinity tolerance. On account of this, the salinity tolerance of the ELRA was tested, assessed with reference substances at several salinity levels, and compared with the E-Screen method and a Yeast Estrogen Screen (YES), which are also frequently applied in environmental testing. The aim of this paper was to explore when the salinity limits within these test procedures are applicable. The trials should reveal the working range to be expected, characterize the salinity-dependent variations in sensitivity of the test, and provide options for methodological adjustments to improve the stability against increased salinity. METHODS: The ELRA was carried out with the human Estrogen Receptor alpha. (ER) using the same principle like a competitive immunoassay based on ligand-protein interaction. However, an essential difference is the use of a physiologically relevant receptor instead of an antibody as a linking protein. The ELRA measures the competition of sample estrogens and anti-estrogens against estradiol supplied as a BSA-coating conjugate for the binding site of dissolved ER. Estradiol or xeno-estrogen binding is quantified by a biotynilated anti-ER antibody and the subsequent measurement of peroxidase activity by a streptavidin-POD-biotin complex. The E-Screen was performed with the human breast cancer cell line MCF-7, which expresses the estrogen receptor constitutively. Cell proliferation depends on binding of estrogens or xeno-estrogens with the receptor. After incubation, estrogen-dependent cell growth was measured by sulforhodamin B staining. The YES was performed with a recombinant yeast strain, transfected with a receptor and a reporter plasmid bearing the estrogen receptor and a vitellogenin gene fused with the reporter gene lacZ. Estrogen or xeno-estrogen-dependent gene induction was measured indirectly by LacZ activity. The salinity levels were simulated in varying concentrations with NaCl from 0 to 40 per thousand or Artificial Sea Water (ASW) from 0 to 32 per thousand. RESULTS: The study characterized the factor 'salinity' for the prospective application fields of the ELRA. With reference substances such as 17-beta-estradiol, the ELRA showed classical sigmoidal concentration-effect relations in a range from 0.05 to 100 microg/l under physiological conditions. After a methodological adjustment to compensate decreasing receptor-binding affinity of estrogens and xeno-estrogens at higher salinity levels, the ELRA became applicable under salinity conditions up to concentrations of 20.5 per thousand. In tests, the ELRA reached under the influence of salinity a mean limit of detection of 0.062 microg/l 17-beta-estradiol. The mean relative inter-test error was around 11%. Above concentrations of 20.5 per thousand there is a risk of false negative assessment. Compared with the E-Screen method using the MCF7 cell line and the yeast estrogen test system (YES), the ELRA shows a lower sensitivity to 17-beta-estradiol. In the E-Screen, the cell proliferation was strongly reduced by sodium chloride induced cytotoxicity. In comparison with the E-Screen, the salinity tolerance of the YES and YAS methods is significantly higher. DISCUSSION: Despite adaption, total salinity tolerance could not be achieved with the ELRA. Freshwater samples were generally appraisable. Higher salinity levels above 20.5 per thousand would tend towards false negative results. The low inter-test error of 11% makes the ELRA suitable for the detection of estrogenic and anti-estrogenic potentials of single substances, substance mixtures, and of environmental samples. CONCLUSIONS: The ELRA is very fast and reproducible, it can be used for high-throughput screening in a microplate format at low cost, it is robust to microbial contamination, and is less susceptible to cytotoxic interferences than cell culture methods. RECOMMENDATIONS AND PERSPECTIVES: In their established form, the YES and the E-Screen methods are not applicable for liquid phase testing at higher salinity conditions. The salinity-adapted test version of the ELRA described here shows a broader working range for samples. Native water samples of more or less brackish origin or high-salinity effluent samples are testable. Results of tests with sediment associated samples of different salinity will be subject of a forthcoming publication.     

成都市区夏季大气挥发性有机物污染特征及来源解析

2019年6~9月在成都市区对挥发性有机物（VOCs）进行在线观测,研究夏季VOCs浓度水平、变化特征、臭氧生成贡献（OFP）及来源贡献.结果表明,成都市区夏季TVOCs（总挥发性有机物）平均质量浓度为112.66 μg·m^-3,烷烃（29.51%）和卤代烃（23.23%）为主要组分;VOCs日变化峰值主要出现在上午10：00~11：00,受城市机动车、油气挥发和工业排放影响;夏季VOCs的OFP贡献中芳香烃贡献率（42.7%）最高,其次为烯烃（27.4%）,关键活性物种为间/对-二甲苯、乙烯、丙烯、邻-二甲苯、异戊烷、环戊烷和丙烯醛等;使用PMF受体模型进行来源解析表明,移动源为成都市区夏季VOCs的主要贡献源,贡献率为34%,其次为工业源（17%）和油气挥发（14%）,溶剂使用源和天然源分别贡献11%和13%.因此,机动车和工业排放为成都市区VOCs的重点控制源,同时溶剂使用及油气挥发等污染源的管控也不可忽视.     

北京城区和郊区大气细粒子有机物污染特征及来源解析

2004年在定陵、北京大学、奥体中心、良乡、通州共5个采样点采集北京市PM2.5,样品,并对其中有机碳(OC)、元素碳(EC)和有机物组成进行了测定.分析了北京市城区和郊区细粒子中有机化合物的污染特征.共检出有机物188种.主要物种为正构烷烃、正构烷酸、霍烷、多环芳烃、脱氧单糖苷以及其它多种源的示踪物.各监测点有机物浓度均呈现1月浓度最高、10月其次,7月浓度最低,4月居中的特征.市区点(奥体、北大)和近郊点(良乡、通州)的污染物浓度远高于受人为活动影响较小的远郊定陵.以正构烷烃、霍烷、多环芳烃、左旋葡聚糖和EC为示踪物,利用化学质量平衡(CMB)受体模型对北京市PM2.5中的OC进行了来源解析.结果表明,北京市细粒子OC的主要来源为柴油车排放(15.3%)、汽油车排放(20.5%)、燃煤排放(19.0%)、生物质燃烧(2.1%)和植物碎屑(1.1%).机动车和燃煤排放仍然是北京市细粒子OC的主要来源.而且有加重趋势.     

50Hz低频环境磁场对细胞增殖的影响及其机理探讨

本文以细胞株LO2(人肝细胞)和CHL(中国仓鼠肺成纤维细胞)为研究对象，用50Hz正弦磁场以不同强度、不同时间作用于细胞。长时间作用后，2种细胞的增殖均受到抑制，但抑制程度与磁场作用量并非呈简单的线性关系；磁场短时间作用对细胞增殖无明显影响，但细胞膜的流动性变差，在磁场撤去后膜的流动性又有所恢复。这一现象说明，低频弱磁场引起宏观生物效应需要时间的积累。同时观察到磁场短时间作用导致的CHL细胞上表皮生长因子受体(EGFR)的聚集，这可能是低频弱磁场引起宏观生物效应的一种机制。     

青藏高原表土重金属污染评价与来源解析

为了解青藏高原表层土壤重金属的污染特征、空间分布及污染来源,沿东北-西南方向对青藏高原表土(0～20 cm)样品进行了采集,对土壤中的Ba、Cd、Co、Cr、Cu、Fe、Mn、Mo、Ni、Pb、Sb、Sc和Zn等13种重金属总量进行了分析,并利用主成分分析-绝对主成分分数-多元线性回归(PCA-APCS-MLR)受体模型初步定量解析了重金属的潜在来源.结果表明Cd和Sb均显著超标,分别是20世纪70年代青藏高原土壤背景值的2. 13与1. 52倍.富集因子(EF)、地累积指数(I_(geo))和Nemero综合指数(PN)分析同样表明青藏高原表土主要以Cd和Sb污染为主,但污染程度普遍不高.在空间分布方面,青藏高原中部、东南部及东北部均呈不同程度污染,但中部及东南部污染相对较重. PCA-APCS-MLR分析表明,青藏高原土壤重金属主要有3个来源,分别为自然、交通和采矿等综合因素. Co、Cr、Cu、Fe、Mn、Ni和Sc主要受自然因素影响,Ba、Cd、Mo和Pb主要受交通因素影响,Zn主要受自然和交通因素共同影响,Sb主要受采矿、自然和交通等综合因素影响.青藏高原土壤重金属污染防治过程中应着重考虑受交通、采矿等综合因素影响下的Cd和Sb污染.     

京津冀水源涵养生态服务供体区与受体区范围的划分

京津冀水资源问题已经严重制约区域经济发展,定量分析研究区水源涵养生态服务供受关系,有助于深刻理解区域上下游之间的生态联系,促进区域生态公平,实现区域社会持续健康发展.利用降水储量法、水量平衡法及水源涵养生态服务供体区与受体区划分方法,分析2000-2010年京津冀净水源涵养量空间格局,划分研究区水源涵养生态服务供体区和受体区范围.结果表明:①冀北燕山山区净水源涵养量最高,年均值为65.24~81.35 mm,其次是冀西太行山山区,年均值为46.47~61.28 mm;净水源涵养量负值区主要分布在东部沿海和大中小城市,年均值为-130.46~-152.39 mm.②水源涵养生态服务供体区占研究区面积的41.42%,单位面积（m2,下同）平均净供水量为0.03~0.07 m3;水源涵养生态服务平衡区面积占25.01%;水源涵养生态服务受体区面积占33.57%,单位面积平均净需水量为0.18~0.41 m3,是供体区供水能力的6倍左右.③从流域来看,滦河流域的承德市为天津市、秦皇岛市及唐山市供应水资源,年均供水总量为23.7×108 m3;永定河流域的张家口市为北京市供应水资源,年均供水总量为5.3×108 m3;大清河流域的保定市为北京市及天津市西南部供应水资源,年均供水总量为8.2×108 m3;但子牙河流域和漳卫河流域供水能力严重不足.研究显示,燕山山区和太行山山区为水源涵养生态服务供体区,东部沿海和大中小城市为主要水源涵养生态服务受体区,水源涵养生态服务供体区净水源涵养能力不能满足水源涵养生态服务受体区需水量,整个研究区供受关系严重失衡.      

大气颗粒物上多环芳烃的识别和源解析的进展

本文对当前大气颗粒物上ＰＡＨｓ的识别和源解析的定量及定性和半定量的方法进行了综述，定性及半定量方法简便易行，但误差大；化学质量平衡法结果较准确具体，但ＰＡＨｓ会发生化学反应而降解，并且没有各种燃烧源较完整的ＰＡＨｓ成份谱，这已成为ＣＭＢ广泛推行的障碍；多元统计方法不考虑ＰＡＨｓ的降解，但要求数据量大，国内外的科学工作者用不同的方法和手段，使ＰＡＨｓ来源的识别和源解析工作有了一定进展。     

工业发达城市土壤重金属时空变异与源解析

采集了宁波市区2003和2013年两期共1 130个土壤样品,测定了8种典型重金属元素的含量,运用污染指数评价法对土壤重金属污染程度进行了评价,运用地统计学方法剖析了污染的时空变异特征,并分别运用主成分分析和UNMIX受体模型解析了重金属的污染来源.结果表明:(1)除了As,两期重金属元素的含量均值都超过了浙江省土壤背景值和国家土壤背景值,说明土壤普遍存在重金属污染;从2003～2013年,所有元素的含量均值都不同程度地增大,说明10年间重金属污染普遍加剧;(2)单因子污染指数的评价结果表明,Hg为重度污染,Cd、Cu、Pb为中度污染,Zn、Cr、Ni为轻度污染,As为非污染状态;所有采样点的内梅罗综合污染指数都超过了警戒值,10年间综合污染指数 3、呈重度污染的样本显著增加.(3)同种元素不同时期的污染空间格局相近,不同元素则呈现一定差异,Pb和Hg,Cr、As、Cu、Zn和Ni的空间分布特征具有相似性,高值区主要集中在污染企业分布密集和人类活动频繁的海曙、江北、江东、镇海四区;(4)研究区重金属污染源主要包括两类,即以Ni、As、Cr、Zn、Cd为主要成分的工业污染源,以Pb和Hg为主要成分的生活污染源,前者源自工业废水、废气和废渣的排放;后者源自于生活垃圾、汽车尾气及道路磨损造成的颗粒物和粉尘污染.