酵母双杂交技术构建稀有鮈鲫不同雌激素受体亚型的重组荧光酵母

目前关于环境雌激素(environmental estrogens,EEs)的效应评估大多数是基于人的雌激素受体(estrogen receptor,ER)的离体检测,缺乏EEs对鱼类ER影响的研究。本研究利用酵母双杂交技术构建模式生物稀有鮈鲫不同ER亚型重组荧光双杂交酵母,用以快速检测EEs的雌激素活性并研究不同ER亚型对17β-雌二醇(E_2)敏感度的差异。采用反转录多聚酶链反应法获取稀有鮈鲫ERα、ERβ1和ERβ2的配体结合域(LBD)序列,构建并鉴别诱饵质粒pGBKT7-ERα-LBD、p GBKT7-ERβ1-LBD和pGBKT7-ERβ2-LBD。将诱饵质粒和猎物质粒pGAD424-GRIP1同时转化荧光酵母Y187-Luc,分别构建3株稀有鮈鲫ERα-GRIP1、ERβ1-GRIP1和ERβ2-GRIP1重组荧光双杂交酵母,并考察了E_2、双氢睾酮(DHT)、9-顺维甲酸(9-cis RA)、三碘甲状腺原氨酸(T_3)和孕酮(PG)对荧光素酶的诱导情况。结果显示:ERα-GRIP1、ERβ1-GRIP1和ERβ2-GRIP1重组荧光双杂交酵母能够专一性地被E_2诱导产生荧光素酶,并存在显著的剂量-效应关系,半数效应浓度(EC_(50))值分别为1.98×10~(-9)、1.77×10~(-10)、和3.52×10~(-10)mol·L~(-1),对E_2的敏感程度排序为:ERβ1-GRIP1>ERβ2-GRIP1>ERα-GRIP1。研究表明,稀有鮈鲫不同ER亚型对E_2的响应具有差异,3株重组荧光双杂交酵母不仅可以应用于快速识别内分泌干扰物中的类雌激素物质,分析稀有鮈鲫不同ER亚型对EEs的敏感性,解析雌激素污染物对稀有鮈鲫的作用机制,评估鱼类接触EEs的潜在风险,以期为水质保障和污染治理提供重要依据。     

Low efficiency of sewage treatment plants due to unskilled operations in India

Centralized sewage treatment plants may not be a sustainable solution for a developing country such as India. Therefore, we conducted for the first time an integrated assessment of the different technologies currently used for sewage treatment in the state of West Bengal, India. Five decentralized sewage treatment plants and one centralized sewage treatment plant located in different parts of Kolkata were evaluated. We compared influent and effluent water quality, energy consumed, capital and operating costs, and treated wastewater reuse potential. F test was used to validate results on the effect of working days and holidays and seasons on treated water quality. Wastewater management strategy was assessed by performance indicators. Our results show that treatment efficiency was lowest in anaerobic plants not because of faulty technology but due to unskilled operation. Therefore, performance improvement of plants is expected if factors such as monitoring, training of staff, regular and scrupulous desludging, reuse aspects, and rational water tariff are implemented earnestly.     

环境雌激素生物效应的作用机制研究进展

环境雌激素(Environmental Estrogens, EEs)种类繁多、来源多样且分布广泛,大量工业添加剂、食品添加剂和农药类物质已被证实具有雌激素活性。EEs对人体生殖、神经、免疫等系统生物毒性,已经引起了公众的普遍关注。近年来的研究表明,EEs不仅可以结合雌激素核受体(nuclear Estrogen Receptor, nER)干扰正常的雌激素基因组信号通路,还能活化雌激素膜受体(membrane Estrogen Receptor, mER)引发快速的雌激素非基因组信号转导途径。本文总结了EEs通过nER、mER介导的多种雌激素基因组和非基因组信号途径及其产生的生物学效应,综述了在其毒理学作用机理基础上发展的环境样品的雌激素活性评估和EEs混合物的联合作用研究,以期为该类污染物的筛查、风险评估和进一步的机制研究提供参考。     

悬浮填料生物反应器处理低浓度氨氮的动力学特性分析

城市污水经一级强化处理后,NH3-N浓度为10~15mg/L,CODCr为40~60mg/L。试验利用悬浮填料床对一级强化处理出水中的氨氮进行深度处理,分析了悬浮填料表面负荷和硝化速率与水力停留时间的关系,并模拟分析了硝化速率与氨氮浓度的关系。结果表明,悬浮填料床内混合液氨氮浓度为0.5~2.5mg/L时,硝化反应符合半级反应动力学,半级反应速率常数k1/2为0.48(g/m)0.5/d;混合液氨氮浓度>2.1mg/L时,硝化反应遵循零级反应动力学,rmax为0.71g/(m2.d)。动力学理论计算值与实际运行结果基本吻合,说明动力学模型对悬浮填料床的硝化性能具有良好的预测和指导意义。     

Field demonstration of surfactant-enhanced solubilization of DNAPL at Dover Air Force Base, Delaware

This study reports on a surfactant-based flood for tetrachloroethylene (PCE) removal from a control test cell at the Dover National Test Site. The surfactant formulation (sodium dihexyl sulfosuccinate (Aerosol-MA or AMA), isopropanol and calcium chloride) was able to achieve a high concentration of PCE in swollen micelles (supersolubilization) without vertical PCE migration. The hydraulic system included eight screened wells that were operated in both vertical circulation and line drive configurations. After 10 pore volumes of flushing, the overall PCE removal was 68% (65% of which corresponded to the surfactant flooding alone). In addition, the residual PCE saturation was reduced from 0.7% to 0.2%, and the concentration of PCE in the groundwater was reduced from 37-190 mg/L before the flushing to 7.3 mg/L after flooding. Recycling the surfactant solution reduced the required surfactant mass (and thus cost, and waste) by 90%. Close to 80% of the total PCE removal was obtained during the first five pore volumes which were operated in an upward vertical circulation flow scheme. No free oil phase was observed during the test. Further analysis of multilevel sampler data suggests that most of the trapped oil remaining in the cell was likely localized in secluded regions of the aquifer, which helps explain the lower PCE groundwater concentration after remedial activities. In summary, this field study demonstrated the feasibility of surfactant-enhanced remediation to reduce the mass in the source zone and significantly reduce the PCE aqueous concentration and therefore the risk associated with the contaminant plume.     

Enhanced Fenton's destruction of non-aqueous phase perchloroethylene in soil systems

The Fenton's system is applied to the destruction of perchloroethylene (PCE) present as a dense non-aqueous phase liquid (DNAPL) in soil slurry systems; the initial concentration of PCE was 45 times higher than its aqueous solubility. Studies were conducted in two matrices: Ottawa sand and soil from Warsaw, IN. In Ottawa sand, a 60-62% decrease in PCE concentration was observed, and Cl(-) recovery was 47-58%, whereas in Warsaw soil, a 44-49% decrease in PCE concentration and a Cl(-) recovery of 40-42% were observed after the addition of 600 mM H(2)O(2) and 10 mM dissolved iron. Significantly enhanced destruction resulted during application of N-(2-hydroxyethyl) iminodiacetic acid (HEIDA) to Warsaw soil. For example, in the absence of HEIDA in Warsaw soil, 36% PCE loss and 33% Cl(-) release were observed at 600 mM H(2)O(2) and 5 mM Fe(III), while 74% PCE loss and 63% Cl(-) release were achieved at 600 mM H(2)O(2) and 5 mM Fe(III)-HEIDA. For both soils, the catalytic activities of Fe(II) and Fe(III) were nearly equivalent. These findings clearly demonstrate that system design can be optimized with regard to process variables in Fenton's treatment of DNAPL in soils.     

In vitro agonistic and antagonistic endocrine disrupting effects of organic extracts from waste water of different treatment processes

Effluents from wastewater treatment plant （WWTP） have been reported to have a broad spectrum of endocrine disrupting compounds （EDCs）. The majority of studies have focused on the occurrence of estrogenic activity, while ignoring nuclear hormone receptors （NRs） pathways. In the present study, a battery of in vitro yeast bioassays and a cell bioassay, including antagonistic and agonistic effects on estrogen receptor （ER）, androgen receptor （AR）, progesterone receptor （PR）, estrogen- related receptor （ERR） and aryl hydrocarbon receptor （AHR）, were conducted to evaluate the removal efficien- cies of EDCs by different treatment processes of a WWTP located in Beijing. Estrogenic, anti-estrogenic, anti- androgenic, anti-progesteronic, anti-ERR and the activa- tion of AHR activities were detected in samples from all treatment processes and the receiving water. The concen- tration of estrogenic contaminants with estradiol （E2） equivalent concentrations ranged from 0.82 x 10-9 to 3.54 x 10 9g Ee_EQ.L-1. The concentration of anti-estrogenic contaminants with 4-hydroxytamoxifen （4-OHT） equiva- lent concentrations ranged from 1.24 × 10-6 to 2.36 x 10-6 g 4-OHT-EQ.L-1. The concentration of anti-androgenic contaminants ranged from 2.21 x 10-s to 3.52 × 10-6g flutamide-EQ. L-1. The concentration of anti-progesteronic contaminants ranged from 3.15 x 10^-5 to 2.71 x 10^-4g RU486-EQ. L-1. The concentration of anti-ERR contami- nants ranged from 7.09 x 10-5 to 6.50 x 104 g 4-OHT-EQ × L^-10. The concentration of AHR activators ranged from 1.7 × 10-10 to 3.4 × 10^-10g TCDD-EQ-L-1. These processes including secondary clarifier, coagulation, as well as coal and sand filtration could eliminated 67.2% of estrogenic contaminants, 47.0% of anti-estrogenic contaminants, 98.3% of anti-androgenic contaminants, 88.4% of anti- progesteronic contaminants, 65.4% of anti-ERR contami- nants and 46.9% of AHR activators. WWTP effluents contain multiple receptor disruptors may have very complex adverse effects on exposed organisms.     

Environmental contamination using accumulation of metals in marine sponge,Sigmadocia fibulata inhabiting the coastal waters of Gulf of Mannar,India

Coastal marine ecosystems in many parts of the world are under unrelenting stress caused by urban development, pollutants and other ecological impacts such as building of infrastructure, land reclamation for port and industrial development, habitat modification, tourism and recreational activities. The present work is a first extensive field study using the marine sponge, Sigmadocia fibulata as a bioindicator to detect metal pollution between inshore and offshore environment of the ‘Gulf of Mannar’, India. Samples of S. fibulata were collected from different places of inshore (0.1–0.5?km) and offshore (3–5?kms) locations. Metal concentrations in water and bioaccumulation in sponges were determined by ICP-MS (Inductively coupled plasma-mass spectrometry). Enrichment of metal contamination was more in the inshore, when compared to offshore environment. Higher concentrations of metals Fe, Al, Ni, Mn, Cu, Cr, Co, Ba, Zn, V, Pb and Cd in inshore waters (3–6.4 times than offshore) may be due to the discharge of monsoonal rains, carrying a discharge of industrial and agricultural wastes and sewage directly into the coast. This is substantiated by a highly significant positive correlation between concentrations of metals in the water and accumulation in sponges. The bioaccumulation of metals in sponge tissue were in order of Fe?>?Al?>?Ni?>?Mn?>?Cu?> Cr?>?Co?>?Ba?>?Zn?>?V?>?Cd?>?Pb in both the inshore and offshore regions. Apparently, the macromolecule composition (sugars, proteins and lipids) was significantly reduced by the accumulation of metals in inshore sponges. The current findings indicated that the frequent monitoring is necessary to assess the eco-health of the marine environment by choosing bio-indicator species like S. fibulata, which provide accurate, reliable measurements of environmental quality.     

Biosorption of chromium(VI), nickel(II) and copper(II) ions from aqueous solutions using Pithophora algae

The biosorption of heavy metals is considered to be one of the best alternatives for the treatment of wastewater. The metal binding capacity of algae and acid-treated algae is investigated to find out the removal characteristics of Cr(VI), Ni(II) and Cu(II) ions from single metal solutions. Batch experiments are conducted and the study is extended to investigate the effect of pH, amount of adsorbent and adsorbate concentration on the extent of biosorption. The results indicate that the adsorption capacity of algae depends strongly on pH. The maximum adsorption of Cr(VI), Ni(II) and Cu(II) occurs at pH values of 2, 7 and 4.3, respectively. The adsorption process follows first-order kinetic equation. The data obtained are correlated with Freundlich and Langmuir adsorption isotherms.     

Effect of nickel exposure on serum amylase activity in nickel-plating workers

The objective of this study was to determine the effect of nickel (Ni) exposure on serum amylase activity in nickel-plating workers in Bangalore (India). Sixty-nine subjects using Ni during the electroplating process formed the exposed group. An equal number of age- and sex-matched subjects working in the administration section formed the control group. Urine Ni levels were determined using a flameless atomic absorption spectrophotometer. Serum amylase activity was determined using spectrophotometric method with 2-chloro-4-nitrophenyl-α-D-maltotrioside as substrate. A significant increase in urine Ni and serum amylase activity was noted in nickel platers as compared with the control group. The level of serum amylase activity was positively and significantly associated with Ni content in urine of nickel platers. Stepwise multiple regression analysis was used to assess the effects of Ni exposure, life style confounding factors and presence of gastrointestinal problems on serum amylase activity. The analysis showed that the subjects who had urine Ni levels beyond 10?µg?g^?1 of creatinine, nickel platers category, smoking and body mass index variables were significantly associated with serum amylase activity. The results of this study suggest that the increased serum amylase activity observed in nickel-exposed subjects could be used as a biomarker for investigating pancreatic function in Ni exposure.