In vitro thyroid disrupting effects of organic extracts from WWTPs in Beijing

It is generally known that there are many endocrine disrupting compounds (EDCs) in the effluents from wastewater treatment plants (WWTPs). Most research has focused on the occurrence of estrogenic or androgenic activities, while ignoring that there are environmental chemicals disrupting thyroid system, which is essential for growth and development in both humans and animals. In the present work, a two-hybrid yeast assay was conducted to evaluate the removal efficiencies of agonistic or antagonistic thyroid receptor (TR) mediated effects in different treatment processes of three WWTPs located in Beijing. We found no TR agonistic, but TR antagonistic activities in all processes from the WWTPs. The TR antagonistic activities in organic extracts of water samples were then calibrated regarding to a known TR-inhibitor, amiodarone hydrochloride (AH). The observed concentration of TR disrupting substances ranged from 2.35 × 10-8 to 6.19 × 10-7 mol/L AH in Gaobeidian WWTP, 3.76 × 10-s to 8.75 x 10-s mol/L AH in Lugouqiao WWTP,and 4.80 × 10-9 to 2.55 × 10-s mol/L AH in Beixiaohe WWTP. Of the three WWTPs, the removal rates were 92.7%, 42.2%, and 23.1% respectively. Industrial sewage may contain more TR disrupting substances compared with domestic sewage. The recipient waters were found to contain considerable concentrations of TR disrupting substances that may cause adverse effects on the exposed organisms.     

Screening of agonistic activities against four nuclear receptors in wastewater treatment plants in Japan using a yeast two-hybrid assay

To assess the potential endocrine disruptive effects through multiple nuclear receptors(NRs),especially non-steroidal NRs,in municipal wastewater,we examined the agonistic activities on four NRs(estrogen receptor α,thyroid hormone receptor α,retinoic acid receptor α and retinoid X receptor α) of untreated and treated wastewater from municipal wastewater treatment plants(WWTPs) in Japan using a yeast two-hybrid assay.Investigation of the influent and effluent of seven WWTPs revealed that agonistic activities...     

Estrogen-related receptor disruption of source water and drinking water treatment processes extracts

Environmental chemicals in drinking water can impact human health through nuclear receptors. Additionally, estrogen-related receptors (ERRs) are vulnerable to endocrine-disrupting e ects. To date, however, ERR disruption of drinking water potency has not been reported. We used ERR two-hybrid yeast assay to screen ERR disrupting activities in a drinking water treatment plant (DWTP) located in north China and in source water from a reservoir, focusing on agonistic, antagonistic, and inverse agonistic activity to 4-hydroxytamoxifen (4-OHT). Water treatment processes in the DWTP consisted of pre-chlorination, coagulation, coal and sand filtration, activated carbon filtration, and secondary chlorination processes. Samples were extracted by solid phase extraction. Results showed that ERR antagonistic activities were found in all sample extracts, but agonistic and inverse agonistic activity to 4-OHT was not found. When calibrated with the toxic equivalent of 4-OHT, antagonistic e uent e ects ranged from 3.4 to 33.1 g/L. In the treatment processes, secondary chlorination was e ective in removing ERR antagonists, but the coagulation process led to significantly increased ERR antagonistic activity. The drinking water treatment processes removed 73.5% of ERR antagonists. To our knowledge, the occurrence of ERR disruption activities on source and drinking water in vitro had not been reported previously. It is vital, therefore, to increase our understanding of ERR disrupting activities in drinking water.     

北京官厅水库水体甲状腺激素干扰效应检测及特征

应用重组人甲状腺激素受体(hTR)基因酵母快速测试方法,检测北京官厅水库水体类/抗甲状腺激素干扰效应,并分析其地理分布、季节分布特征以及水体甲状腺激素干扰化合物(TDCs)组成特征.结果表明:官厅水库水体检出了显著的抗甲状腺激素干扰效应,与其它研究区域相比,效应处于较低到中等水平;入库河流对库区水质产生显著影响,且入库河流水质受季节影响显著,因此,库区水体不同季节甲状腺激素干扰水平存在差异;C18固相萃取柱对抗甲状腺激素干扰效应的去除率在61.9%~92.3%,表明官厅水库水体中的甲状腺激素抑制活性物质多集中在有机组分,加入30mg/L的EDTA后TR抑制活性显著降低,表明官厅水库水体中TDCs除有机化合物外可能还包含重金属类无机化合物.     

改进型重组基因酵母TR-GRIP1检测化合物甲状腺激素干扰活性

应用酵母双杂交技术构建重组TR(甲状腺激素受体)基因酵母,用以检测类/抗甲状腺激素化合物及环境样品的甲状腺激素干扰活性. 提取并纯化含有TR基因的酵母表达质粒pGBT9-TR以及含有TR共激活因子GRIP1基因的酵母表达质粒pGAD424-GRIP1,将pGBT9-TR和pGAD424-GRIP1同时转化至酵母细胞Y187,以营养缺陷型培养基(SD/-Trp/-Leu)筛选阳性菌落,构建TR-GRIP1双杂交酵母. 考察该酵母与天然甲状腺激素——T₃(三碘甲状腺原氨酸)的结合情况,确定最佳暴露时间,建立剂量－效应关系曲线. 结果表明:TR-GRIP1双杂交酵母能够与T₃结合诱导β-半乳糖苷酶活性,选择暴露时间为2 h,T₃诱导酶活性的EC₅₀值为1.1×10-7 mol/L;构建的重组基因酵母TR-GRIP1提供了检测化合物甲状腺激素干扰活性的新方法.      

Profile and removal of endocrine disrupting chemicals by using an ER/AR competitive ligand binding assay and chemical analyses

An estrogen receptor (ER)/androgen receptor (AR) ligand competitive binding assay (ER/AR-binding assay) and chemical analyses were used to evaluate the endocrine disrupting chemicals (EDCs) behavior of two municipal wastewater treatment plants (WWTPs) (K and S). In the influents, estrone (E1), androsterone (A), androstenedione (AD), BPA (bisphenol A), NP (nonylphenol) and daidzein (DZ) were detected in high amounts with subsequent 24 h-average concentrations of 350, 1000, 29, 1300, 3900, and 5700 ng/L in K-WWTP and of 310, 620, 59, 1600, 2600, and 8400 ng/L in S-WWTP. The estrogenic (androgenic) activity as 17 -estradiol (E2) equivalents (EEQ) or testosterone (Te) equivalents (TEQ) was consequently 620 ng E2/L (570 ng Te/L) and 580 ng E2/L (800 ng Te/L) for the two WWTPs. The removal e ciencies of the above mentioned sole target chemicals were 51%–100% for K-WWTP and 55.6%–100% for S-WWTP. The removal e ciencies of EEQ were about 73% for both WWTPs, while the removal e ciencies of TEQ were 62.1% for K-WWTP and 98.4% for S-WWTP. In addition, chemical-derived EEQ were about 1.2%–52.4% of those by ER-binding assay for K-WWTP and the corresponding ratios were 1.3%–83.3% for S-WWTP, while chemical derived TEQ were less than 3% of values measured by the AR-binding assay for both WWTPs.     

精神活性物质在北京市某污水处理厂中的污染特征与生态风险

为了解污水处理厂对精神活性物质的去除特征及总出水对受纳水体的生态风险,采用固相萃取-高效液相色谱-串联质谱法调查了北京市某污水处理厂中13种精神活性物质的浓度水平与负荷量变化,并运用RQ（risk quotient,风险熵）对总出水中精神活性物质进行风险评估.结果表明:①13种精神活性物质在总进水与总出水中均能检出,总质量浓度平均值分别为2 395.10和63.59 ng/L,其中ρ（EPH）（EPH表示麻黄碱）占比分别为93.9%和67.9%,其次为COD（可待因）与METH（甲基苯丙胺）.污水处理厂上游地表水中ρ（EPH）、ρ（METH）与ρ（KET）（KET为氯胺酮）均高于总出水及其下游地表水,说明上游沿河可能有新的污染源输入.②污水处理厂对NK（去甲氯胺酮）、BE（苯甲酰爱康宁）和MTD（美沙酮）均呈负去除,其他精神活性物质的去除主要发生在二级生物处理与三级处理（超滤膜与UV消毒）阶段.③污水处理厂服务区域内精神活性物质的周内负荷量存在一定波动,AMP（苯丙胺）、METH、MDA（3,4-亚甲二氧基苯丙胺）、MDMA（3,4-亚甲二氧基甲基苯丙胺）、KET与HER（海洛因）的负荷量均在周末升高.④污水处理厂总出水中精神活性物质的生态风险均较低（RQ < 0.10）.研究显示,污水处理厂不能完全去除污水中的精神活性物质,总出水中残留精神活性物质对受纳河流生态系统产生的长期混合效应不容忽视.      

Degradation of dissolved organic matter in effluent of municipal wastewater plant by a combined tidal and subsurface flow constructed wetland

Dissolved organic matter (DOM) is an important constituent of wastewater treatment plant (WWTP) effluent. A novel combined tidal and subsurface flow constructed wetland (TF-SSF-CW) of 90 L was constructed for a ten-month trial of advanced treatment of the WWTP effluent. Excitation emission matrix (EEM) fluorescence spectroscopy, parallel factor (PARAFAC) analysis and a two end-member mixing model were employed to characterize the composition and removal process of the effluent DOM (EfOM) from the WWTP. The results showed that the TF-SSF-CW performed an efficient EfOM removal with dissolved organic carbon (DOC) removal rate of 88% and dissolved organic nitrogen (DON) removal rate of 91%. Further analysis demonstrated that the EfOM consisted mainly of two protein moieties and two humic-like groups; protein moieties (76%) constituted the main content of EfOM in raw water and humic-like groups (57%) became the dominating contributor after treatment. The EfOM from the WWTP was mainly of aquatic bacterial origin and evolved to a higher proportion of terrigenous origin with higher humification in the TF-SSF-CW effluent. A common controlling treatment-related factor for determining the concentrations of the same kind of substances (protein groups or humic-like groups) was revealed to exist, and the ratio of removal rates between the same substances in treatment was calculated. Our study demonstrates that the TF-SSF-CW can be a novel and effective treatment method for the EfOM from WWTPs, and is helpful for understanding of the character of EfOM in wetland treatment.     

低碳氮比进水AAO污水处理厂低碳运行

为探明低碳氮比进水条件下AAO污水处理厂的碳排放特征,提出可行的低碳运行策略,基于排放因子法对7座低碳氮比进水AAO污水处理厂(分为AAO组和AAO-MBR组)运行1a产生的碳排放进行核算与评价,对具有显著低碳表现的污水处理厂开展全流程分析剖析其碳减排途径。结果表明,电耗和N₂O排放是主要碳排放来源,分别贡献49.43%和25.75%的碳排放。AAO-MBR组以间接碳排放为主,电耗碳排放占至约60%,而AAO组生物作用导致的直接碳排放占主导。AAO组平均吨水比碳排放显著低于AAO-MBR组(0.47和0.79kgCO_2eq/m³),更具低碳运行潜力。7座污水处理厂中,WWTP7各项比碳排放评价指标均为最低,意味着其最具低碳运行能力。充分利用进水碳源,多路径协同脱氮除磷同时精准控制溶解氧浓度避免过曝气是其大幅削减能耗和物耗,实现碳减排的关键路径。     

Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR)

The detection of viable bacteria in wastewater treatment plants(WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide(PMA) combined with the quantitative polymerase chain reaction(PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 μmol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.     

污水处理厂中微塑料的去除效能与全流程分析——以北京某下沉式三级污水处理厂为例

污水处理厂排放是微塑料进入自然环境的重要途径之一,本研究对北京某下沉式三级污水处理厂八个工艺单元中微塑料的赋存特征和去除效能进行全流程分析.研究表明该厂赋存微塑料主要形态为纤维,主要成分为聚丙烯、聚酯和聚乙烯,尺寸≤500μm的微塑料影响最为显著.该厂对微塑料的整体去除率为91.7%,主要依靠沉淀和截留过滤去除.双层平流沉淀池（二沉池）的去除效能最高,而生化处理单元没有明显去除效果.整体上,微塑料呈现从污水向污泥迁移的趋势.该厂最终出水中微塑料浓度为1.3n/L,成分为聚酯纤维,出水排放没有引起周边受纳水体中微塑料浓度升高,但干扰了下游地表水体中微塑料的成分,深度处理单元是降低受纳水体微塑料污染风险的重要保障.     

A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

Amphibian metamorphosis provides a wonderful model to study the thyroid hormone (TH) signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A (TBBPA), a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3, whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.     

基于2D-COS分析的难凝聚有机物臭氧化特性研究

基于二维相关光谱分析,对污水厂二级出水难凝聚有机物的臭氧化特性进行了研究.结果表明,混凝很难去除污水厂二级出水中的溶解性有机物,污水厂二级出水溶解性有机物大部分属于难凝聚有机物,以腐殖酸类物质为主.臭氧氧化可以降低二级出水混凝后上清液的色度及UV₂₅₄,去除率分别可达到70%和40%以上,但是对难凝聚有机物的去除率较低,去除率在10%以下,只是与难凝聚有机物进行了反应,改变其性质.同时,臭氧可以降低二级出水中难凝聚有机物的荧光强度,其中腐殖酸类物质的荧光强度降低较为明显.基于不同臭氧投加量下难凝聚有机物的同步荧光分析,可以得到二级出水难凝聚有机物的二维相关光谱,结果表明,腐殖酸类物质对臭氧氧化最为敏感,腐殖酸类物质与富里酸类物质相比会优先和臭氧反应.     

城市污水处理过程中微塑料赋存特征

以上海某污水厂为例,研究了中国沿海城市污水厂进出水、污泥中微塑料的赋存特征及微塑料排放量.结果表明,污水厂进出水中微塑料形态主要为纤维状.进水含量最高的种类依次为人造丝（36.84%）、合成革（21.05%）和聚酯纤维（18.42%）,出水中的种类依次为合成革（26.67%）、人造丝（24.44%）、聚酯纤维（17.78%）和聚乙烯（15.56%）.微塑料在总进水中的丰度为117n/L（n为微塑料个数）,调配水中为90n/L,总出水中为52n/L,污泥中为180n/50g（湿重）.该污水厂日出水量为280万m³/d,据此估算微塑料量由总进水中3276亿n/d降至总出水中1456亿n/d,去除率为55.6%.污水厂根据实际进水水质确定剩余污泥量150tDS/d,含有5.4亿个微塑料.此外,根据本研究结果讨论了有关国际刊物中对我国大陆范围内个人护理用品中微塑料入海量的不准确估算.     

基于全流程分析的低碳氮比进水污水处理厂运行调控

为使某污水处理厂出水达标排放,对该厂进行了全流程测试,分析其主要污染物沿工艺流程分布特征以及活性污泥特性,评估工艺运行现状,为该污水处理厂优化调控提供基础数据。研究发现,该厂进水ρ（BOD₅）/ρ（TN）仅为2.45,属于典型的低碳氮比进水。此外,通过活性污泥特性测试发现,反硝化潜力为9.0 mg/（g·h）,反硝化菌群相对丰度较高。进水碳源不足及外部碳源投加位点设置不合理是该厂无法实现TN达标排放的主要原因。在采取改变碳源投加位点、减小好氧池末端曝气量、增加碳源投加量等措施后,出水ρ（TN）由32.0 mg/L降至12.7 mg/L,实现了TN的达标排放;此外,厌氧释磷潜力由1.3 mg/（g·h）提升至2.6 mg/（g·h）,生物除磷能力也有了较大提升。研究提供了一种解决污水处理厂出水水质超标问题的思路,可为含低碳氮比进水的城镇污水处理厂运行调控及稳定达标提供参考。     

基于达标保证率的昆明市污水处理厂出水水质评价

采用多元统计分析,将水质分级评价法和达标保证率法相结合,评价了昆明市7座污水处理厂出水水质.以《城镇污水处理厂污染物排放标准》(GB18918-2002)一级A限值为达标基准,7座污水处理厂出水COD达标保证率均为100%、出水BOD5和SS达标保证率分别在94%和95%以上,不同污水处理厂出水TP、NH3-N和TN的达标保证率表现出较大差异,多维标度分析表明,出水TP的达标保证率与其他指标的偏离程度最大.将污染指标的达标保证率进行分级,建立污水处理厂污染指标达标满意程度评价规则,并制定出水水质的达标控制指标清单.结果表明,TP是第二和第七污水处理厂的重点控制指标,是第三污水处理厂的Ⅰ级控制指标;TN是第三和第七污水处理厂的Ⅰ级控制指标;NH3-N是第一和第三污水处理厂的Ⅰ级控制指标.     

有机磷阻燃剂甲状腺干扰效应及其作用机制研究——以磷酸三(2-氯丙基)酯(TCPP)为例

以典型有机磷阻燃剂磷酸三（2-氯丙基）酯（TCPP）为研究对象,应用GH3细胞增殖实验检测TCPP对甲状腺激素的干扰效应;应用重组甲状腺激素受体（TR）基因酵母实验和GH3（TRβ-）细胞增殖实验结合实时定量PCR技术初步探究TCPP甲状腺激素干扰作用机理.结果表明,TCPP在1×10^-4mol/L和2×10^-4mol/L浓度下对甲状腺激素T3诱导的GH3细胞增殖产生抑制效应;重组TR基因酵母实验和GH3（TRβ-）细胞增殖实验测试结果表明TCPP可能通过TR介导的基因组途径和非基因组途径诱导甲状腺激素干扰效应;实时定量PCR测试结果表明TCPP下调相关基因如：c-fos、TRβ、integrin-av和k-ras的mRNA表达,初步认为TCPP可能通过影响TRβ基因表达和激活α_vβ₃-ERK-1/2信号通路产生甲状腺激素干扰效应.     

大型城市污水处理厂处理工艺对微塑料的去除

污水处理厂出水是自然水体中微塑料(MPs)的重要来源.本文以上海市两个大型污水处理厂(WWTP)为研究对象,分析了进水及各处理工艺出水中MPs的数浓度、形态变化及相应的去除率差异,计算了MPs在污水处理厂的归趋.结果表明,WWTP1和WWTP2进水中MPs数浓度分别为(226. 27±83. 00)个·L-1和(171. 89±62. 98)个·L-1; WWTP1对MPs的去除效率为63. 25%,略大于WWTP2的处理效率(59. 84%);两个污水处理厂一级处理工艺对MPs的去除率占整个处理工艺的70%～80%.从污水处理厂的处理工艺来看,一级处理工艺和二级处理工艺分别将污水中(48. 10%±1. 62%)和(12. 97%±0. 05%)的MPs转移到污泥中.从整体来看,污水处理厂中的最终有(38. 82%±1. 55%)的MPs随出水进入到自然水体,剩余(61. 18%±1. 55%)的MPs进入到污泥中.本研究表明,上海市污水处理厂对MPs的去除率较低,即使在处理后依旧有大量的MPs随出水进入到自然水体,仍会对生态系统造成巨大的风险.本研究提供了平原河网地区大型城市污水处理厂MPs去除及归趋的基础数据,可为进一步的MPs去除工艺设计提供参考.     

不同微生物处理工艺对全氟化合物的去除效果

以辽宁省4个采用不同微生物处理工艺的污水处理厂为研究对象,采用WAX固相萃取分离富集和高效液相-质谱联用(HPLC/MS/MS)方法测定了4个污水处理厂中13种PFCs(全氟化合物)〔9种PFCAs(全氟羧酸类化合物,C₄~C₁₂)和4种PFSAs(全氟磺酸类化合物,C₄、C₆、C₈、C₁₀)〕的质量浓度. 研究发现,9种PFCAs(C₄~C₁₂)(记为∑PFCAs)和2种PFSAs(C₄、C₈)(记为∑PFSAs)均有不同程度的检出,水体中各PFCs浓度水平与碳链长度呈显著负相关. 进水中ρ(∑PFCAs)和ρ(∑PFSAs)分别为81.5~135.9和3.7~4.6 ng/L,出水中ρ(∑PFCAs)和ρ(∑PFSAs)分别为47.4~63.3和2.8~3.7 ng/L. 进、出水受PFCAs尤其是短链PFCAs污染较为严重. 4个污水处理厂对PFCs的去除呈现相似的去除效果及规律,即PFCs的去除主要靠活性污泥的吸附作用,而与污水处理厂所采用的微生物处理工艺相关性不大.      

北京市清河维A酸受体活性调查及原因物质解析

为了研究水环境中维A酸受体活性的成因,于2009年3~7月对清河河道进行了4次采样分析,利用固相萃取柱富集与重组基因酵母结合测试方法,研究了污水排放口上下游河段水样RAR活性的时空变化,并利用高效液相色谱分割活性馏分对原因物质进行了解析.结果发现,清河污水处理厂的上下游部分河段水样具有较高的RARα激活活性,但活性地点位置不稳定,随时间变化.同时,在对原因物质的解析中发现,all-trans-4-oxo-RA和13-cis-4-oxo-RA只是极小部分原因物质,环境水样中依然存在大量的未知RAR活性物质.