Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.     

一体式膜生物反应器污水处理工艺研究

在一体式膜生物反应器研制开发实验及反应器各参数运行协调理论分析的基础上,提出了一体式膜生物反应器工艺各参数间的设计原则、方法、步骤及设计实例,对该工艺的运行能耗作了评价.     

A novel approach to treat combined domestic wastewater and excess sludge in MBR

Domestic wastewater was treated by combined anaerobic biofilm-aerobic membrane bioreactor(MBR) process, and part biomass in MBR was withdrawn, to treat with ozone, then the ozonated sludge was returned to anaerobic inlet. In aerobic MBR, MLSS and DO were controlled at 3000—3500 mg/L and 0.8 mg/L respectively. Comparing the experimental results of two stages, it was noticed that ozonation did not affect the removal efficiency for organics but had a significant influence on the removals of NHs-N and TN. During the ozonation period of two months, no excess sludge was wasted, and a zero sludge yield was obtained.     

中水回用工程技术——膜生物反应器

抚顺市肉联厂的中水回用工程每日处理中水100m^3，采用MBR工艺。经过试运行后，CODCr去除率为51％，BOD5去除率为77％，悬浮物去除率为93％。实践证明，MBR工艺是一种简易、高效的中水回用技术。     

应用投加粉末活性炭的膜生物反应器处理生活污水的研究

应用向间歇曝气的膜生物反应器内投加粉末活性炭的新工艺，进行了处理模拟生活污水的研究，将此研究结果与未投加粉末活性炭时的研究结果相比较，表明该工艺不仅可以取得更优的出水水质，而且可以从根本上减少膜阻力，维持高膜通量。     

影响MBR脱氮效率的因素研究

采用重力出流式膜生物反应器,考察了采用2种脱氮运行方式对生活污水脱氮的效果,同时对影响脱氮效率的相关因素进行了分析．研究结果表明,C／N是影响A／O与同步硝化反硝化（SND）2种运行方式脱氮效率最关键的因素．当C／N比小于12时,A／O方式的脱氮效率高于SND;随着C／N的升高,A／O与SND脱氮效率的差距逐渐缩小．对于A／O运行方式,在低C／N条件下可以通过提高回流比与增加缺氧段的HRT来提高系统的总氮去除率．而对于SND运行方式,好氧反硝化的完成是建立在有足够有机物作碳源的基础上,此时,DO的控制就成为提高脱氮效率的关键因素．     

二苯碳酰二肼分光光度法测定水中六价铬样品预处理方法的改进

本文对二苯碳酰二肼分光光度法测定地表水及工业废水中六价铬的样品预处理方法提出了改进措施。改进方法准、快、简、省,符合环境监测分析要求。用该方法对样品进行预处理后分析,其结果与文献[1]一致。     

酱油废水处理技术初探

酱油废水属于比较难处理的高浓度有机废水 ,色度去除更是一个难题 ,从物化和生化处理角度探讨酱油废水处理技术 ,实例表明COD、BOD去除率达 8% ,有待研究。     

PAC-MBR处理垃圾渗滤液的硝化性能研究

研究了粉末活性炭(PAC)作为载体的膜生物反应器(MBR)处理吹脱后垃圾渗滤液的硝化性能.在HRT=3 d、NH 4-N=200～500 mg/L条件下,进水pH＞8.7时,PAC-MBR和没有添加PAC的普通MBR均只能将氨氮转化为NO-2;降低pH至7.6～8.2时,PAC-MBR中的亚硝酸盐氧化菌迅速恢复活性,将NO-2完全转化为硝酸盐,而普通MBR仍然停留在亚硝化阶段;当吹脱取消后,NO-2迅速消失,这可能与取消吹脱后,不再使用大量硫酸进行pH调节有关.研究发现,当SO2-4为3 g/L时,氨氮氧化速度出现了明显的下降,表明SO2-4对硝化菌具有抑制作用;同时,微生物代谢产物(SMP)分泌量越高,NO-2含量越低,但是SMP不是影响NO-2积累的主要原因.     

膜生物反应器中膜分离单元运行参数优化试验

用生活杂排水对膜生物反应器中膜分离单元的运行参数进行了优化试验研究，膜分离单元采用中空纤维超滤膜，试验表明，在适宜的反冲洗和化学清洗条件下，膜的透水率可及时、有效地恢复，使膜生物反应器能长期稳定地运行。