村落耕作型稻田湿地中类固醇类激素HD菌强化降解试验

农村生活污水中的类固醇类激素（EDCs）通过灌溉形式直接被稻、麦、瓜果等农作物吸收,进而进入食物链,带来的环境污染及生态危害问题日益突出。本研究以水稻田为载体构建耕作型稻田湿地,并以本课题组筛选的农药降解菌HD为EDCs强化降解菌,考察耕作型稻田湿地对EDCs的去除效率。试验设A（空白对照）、B（投加HD菌剂强化）两组,结果表明,A组对雌酮（E₁）、雌二醇（E₂）、雌三醇（E₃）及17α-乙炔基雌二醇（EE₂）的平均去除率分别为80.1%、72.4%、51.4%、68.3%,B组平均去除率分别为82.6%、73.4%、60.5%、75.6%,除E₃外E₁、E₂、EE₂去除率均在65%以上。经HD菌剂强化后,耕作层、土壤层、填料层等3个生物单元对EDCs（E₁、E₂、E₃、EE₂）的去除效果均有提升,其中对E₁去除率分别为16.2%、75.1%、28.2%;对E₂去除率分别为14.5%、61.8%、18.7%;对E₃去除率分别为20.6%、49.8%、10.4%;对EE₂去除率分别为8.9%、43.4%、-8.1%。湿地经过5个月的连续运行后,EDCs含量下降明显,投菌组B中的EDCs残留量明显低于未投菌组A,B组湿地出水中E₁、E₂、E₃和EE₂含量相比A组可分别降低约26.6%、17.1%、30.3%、13.3%。HD菌剂能有效强化村落耕作型稻田湿地对EDCs的降解。     

Assessment of a potential preventive ability of amygdalin in mycotoxin-induced ovarian toxicity

The possible effects of a natural substance amygdalin and its combination with the mycotoxin deoxynivalenol (DON) on the steroid hormone secretion (progesterone and 17-β-estradiol) by porcine ovarian granulosa cells (GCs) were examined in this in vitro study. Ovarian GCs were incubated without (control group) and with amygdalin (1, 10, 100, 1,000 and 10,000 μg mL¹), or its combination with DON (1 μg mL¹) for 24 h. The release of steroid hormones was determined by ELISA. The progesterone secretion by porcine ovarian GCs was not affected by amygdalin in comparison to the control. However, the highest amygdalin dose (10,000 μg mL¹) caused a significant stimulation of the 17-β-estradiol release. A combination of amygdalin with DON significantly (P < 0.05) increased the progesterone release at all concentrations. Similarly, a stimulatory effect of amygdalin co-administered with DON was detected with respect to the 17-β-estradiol secretion at the highest dose (10,000 μg mL¹) of amygdalin and 1 μg mL¹ of DON. Noticeable differences between the effects of amygdalin alone and its combination with DON on the progesterone release were detected. In contrast, no differences between the stimulatory effects of amygdalin and its combination with DON on the 17-β-estradiol synthesis by porcine GCs were observed. Findings from this in vitro study did not confirm the expected protective effect of amygdalin on mycotoxin induced reprotoxicity. Our results indicate that the stimulatory effect of amygdalin combined with DON on the progesterone release was clearly caused by the DON addition, not by the presence amygdalin per se. On the other hand, the stimulation of 17-β-estradiol production was solely caused by the presence of amygdalin addition. These findings suggest a possible involvement of both natural substances into the processes of steroidogenesis and appear to be endocrine modulators of porcine ovaries.     

The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

我国直辖市畜禽养殖排放类固醇激素特征及其潜在污染风险

通过对北京、天津、重庆及上海4个直辖市畜禽养殖的统计分析,借鉴英美等发达国家的相关研究,估算了各直辖市畜禽养殖业类固醇雌激素和雄激素的年排放量,并初步探讨了各市单位耕地面积的类固醇雌激素承载量及类固醇雌激素对地表水体的潜在污染风险.结果表明,4个直辖中,重庆市的雌激素和雄激素年排放量处于4个直辖市之首,2008年两类激素的排放量分别达到1283.0和136.1 kg.北京和上海市类固醇激素总排放量均较10年前有所减少,尤其是上海市,10年间类固醇激素排放量减少了1/3.相反,天津市和重庆市的畜禽养殖业类固醇激素排放量却在增长.北京市单位耕地面积畜禽粪便类固醇雌激素的承载量最大,且明显增大,2008年达到839.1 mg·hm-2,是1998年的1.5倍.根据英国环保署(Environment Agency,United Kingdom)建议的最低可观测效应浓度10 ng·L-1,北京市和天津市的畜禽养殖排泄物有可能对当地地表水系统产生生态危害.在3种类固醇雌激素中,雌酮的排放对耕地承载的贡献最大.因此,各直辖市尤其是北京市和天津市亟需加强规模化养殖业管理,加强畜禽养殖业排污中类固醇激素的去除,保障城市的耕地及地表水生态安全.     

SEs在城市污泥处理处置过程中的归趋研究进展

近年来,内分泌干扰物因损坏生物的内分泌系统而引起人们的广泛关注。其中甾体雌激素(SEs)由于具有极强的内分泌干扰能力,在极其微量的情况下便可对生态系统及人和动物的健康产生不可预测的危害,而城市污水处理厂排放的剩余污泥是环境中该类化合物的重要来源。文章结合最近国内外的研究成果,指出了研究内分泌干扰物在城市污水处理厂剩余污泥处理过程中归趋的重要性,重点总结了有关雌酮、17β-雌二醇、雌三醇、17α-乙炔雌二醇等甾体雌激素在城市污水处理厂污泥处理过程中归趋的主要研究进展,包括4种甾体雌激素的基本理化性质、城市污泥处理系统各处理单元中雌激素的背景含量、主要污泥处理过程对雌激素的去除效果及其机制等,并展望了今后该方面的研究前景、热点和趋势。     

基于响应面法优化污水中雌酮的固相萃取条件

为提高污水样品中雌酮(E1)的固相萃取回收率,应用响应面法(RSM)对影响固相萃取的关键参数进行了优化,建立了固相萃取回收率的二次多项式模型,分析了模型有效性和因子交互作用,确定了最佳固相萃取条件;并对实际污水处理厂进、出水样品中的E1进行了固相萃取和浓度检测。结果表明,影响E1固相萃取回收率的因素重要性依次为:洗脱体积>进样速率>洗脱速率。最佳固相萃取条件为:洗脱体积11.15 mL;进样速率10.62mL/min;洗脱速率4.15 mL/min;在此条件下,预测回收率最大可达81.82%。分别采用SBR、氧化沟和A2/O工艺的3座污水处理厂进水E1浓度分别为36.8⁸9.0、24.189.0、24.1²8.4和27.828.4和27.8⁵8.1 ng/L,对应去除率分别为62.8%58.1 ng/L,对应去除率分别为62.8%⁷7.0%、49.3%77.0%、49.3%⁶3.6%和56.1%63.6%和56.1%⁷4.9%。3种污水处理工艺对E1均有一定的去除能力,但出水中残余E1仍远超过预测无效应浓度。     

天然类固醇雌激素源解析、环境行为及其污染控制

天然类固醇雌激素是一种在生物体内具有极强雌激素效应的内分泌干扰物,其在环境中的广泛存在严重影响野生动物尤其是鱼类的正常生长和发育,对环境生态系统所产生的破坏也日趋严重,然而其危害依然没有引起足够的重视。文章研究在阐述天然类固醇雌激素基本理化性质和环境存在形态的基础上,解析其在人类和畜禽的环境来源,分析其在地表水和地下水的污染状况以及环境迁移和生物积累等环境行为,并综合探讨了现今较为常见的如活性污泥法、光降解法以及微滤、纳滤和反渗透等与天然类固醇雌激素相关的污染控制技术,以期为天然类固醇雌激素的毒理性质、污染现状及相关控制方法的深入研究提供支撑。在此基础上,提出深入天然类固醇雌激素微量检测技术研究、加强国家行政管理层面上的污染控制等已十分急迫。     

再生水中微量有毒有害污染物的初步筛查

采用气相色谱质谱联用(GC-MS)、电感耦合等离子体质谱(ICP-MS)等方法对5个污水处理厂再生水中的162种微量有毒有害污染物进行了评价。在各种污染物中,酚类、酞酸酯、类固醇激素和金属类物质被检出。酚类物质的质量浓度为0.01~148 ng/L,符合地表水环境质量标准,但三氯生、双酚A仍存在一定的生态风险;酞酸酯类物质的质量浓度为0.1~0.45μg/L,符合地表水环境质量标准;类固醇激素的质量浓度为0.5~30 ng/L,具有内分泌干扰效应,需优先控制;金属类物质的质量浓度为1~27μg/L,满足污水排放标准和地表水环境质量标准。     

Associations between maternal phthalate exposure and cord sex hormones in human infants

It has been speculated that maternal phthalate exposure may affect reproductive development in human newborns. However, the mechanism awaits further investigation. The aim is to evaluate the association between maternal phthalate exposure and cord sex steroid hormones in pregnant women and their newborns from the general population. A total of 155 maternal and infant pair were recruited and analyzed. Levels of urinary phthalate metabolites and sex steroid hormones were determined using liquid chromatography/electrospray tandem mass spectrometry (LC-ESI-MS/MS) and radioimmunoassay (RIA), respectively. No significant correlation was found between each steroid hormones and phthalate metabolites for male newborns, except MMP was marginally significantly correlated with E₂. After adjusting for maternal age, estradiol (E₂) levels in cord serum from male newborns were not correlated with maternal urinary phthalate metabolites. In female newborns, the maternal urinary levels of mono-(2-ethylhexyl) phthalate (MEHP) and mono-(2-ethyl-5-hydroxyhexyl) phthalate (5OH-MEHP) were negatively correlated with the free testosterone (fT) and fT/E₂ levels in cord serum with Pearson correlation coefficients ranging between −0.24 and −0.29 (p < 0.05). Additionally, after gestational age was adjusted, the maternal urinary level of DEHP was negatively correlated with the free testosterone (fT) and fT/E₂ levels in cord serum. We suggest that maternal exposure to phthalates may affect sex steroid hormones status in fetal and newborn stage.