Hepatic and extrahepatic expression of estrogen-responsive genes in male adult zebrafish (Danio rerio) as biomarkers of short-term exposure to 17β-estradiol

Growing concern over reproductive hormones in the environment demands sensitive and efficient methods by use of molecular biomarkers to detect these contaminants in oviparous vertebrates. In this study, a real-time quantitative RT-PCR was adopted to investigate the expressions of three estrogen-responsive genes, Vtg I, Vtg II and ERalpha, in hepatic and extrahepatic tissues of male adult zebrafish exposed to varying concentrations of 17beta-estradiol (E2) for selected periods. Without exposure to E2, all the genes were expressed in the tissues of male controls with their levels being much lower than those in the respective tissues of female controls. The expressions of hepatic Vtg I and II mRNAs were induced significantly after 1-day exposure to E2 at as low as 0.25 microg L(-1). Significant induction in the expression of hepatic ERalpha mRNA required a higher E2 concentration (> or = 0.5 microg L(-1)) and a longer exposure (> or = 2 days), suggesting that Vtg I and Vtg II are more sensitive to E2 exposure. The induction of Vtg mRNA in the skin of zebrafish was also significant following a short exposure (1 day) to low E2 concentration (0.25 microg L(-1)), with the levels of Vtg I and Vtg II mRNA being increased by 25 and 5 times, respectively. These results suggest that Vtg I mRNA is a highly sensitive biomarker for determining the estrogenic effects of E2 and that the skin of zebrafish may be an appropriate substitute for liver for such a determination.     

Indexing method for assessment of pollution potential of leachate from non-engineered landfill sites and its effect on ground water quality

Dumping of solid waste in a non-engineered landfill site often leads to contamination of ground water due to leachate percolation into ground water. The present paper assesses the pollution potential of leachate generated from three non-engineered landfill sites located in the Tricity region (one each in cities of Chandigarh, Mohali and Panchkula) of Northern India and its possible effects of contamination of groundwater. Analysis of physico-chemical properties of leachate from all the three landfill sites and the surrounding groundwater samples from five different downwind distances from each of the landfill sites were collected and tested to determine the leachate pollution index (LPI) and the water quality index (WQI). The Leachate Pollution Index values of 26.1, 27 and 27.8 respectively for landfill sites of Chandigarh (CHD), Mohali (MOH) and Panchkula (PKL) cities showed that the leachate generated are contaminated. The average pH values of the leachate samples over the sampling period (9.2 for CHD, 8.97 for MOH and 8.9 for PKL) show an alkaline nature indicating that all the three landfill sites could be classified as mature to old stage. The WQI calculated over the different downwind distances from the contamination sites showed that the quality of the groundwater improved with an increase in the downwind distance. Principal component analysis (PCA) carried out established major components mainly from natural and anthropogenic sources with cumulative variance of 88% for Chandigarh, 87.1% for Mohali and 87.8% for Panchkula. Hierarchical cluster analysis (HCA) identifies three distinct cluster types for the groundwater samples. These clusters corresponds to a relatively low pollution, moderate pollution and high pollution regions. It is suggested that all the three non-engineered landfill sites be converted to engineered landfill sites to prevent groundwater contamination and also new sites be considered for construction of these engineered landfill sites as the present dumpsites are nearing the end of their lifespan capacity.     

Evaluation of an Exposure Assay to Measure Uptake of Sediment Pah by Fish

The ecological risks of polynuclear aromatic hydrocarbons (PAH) in aquatic sediments will vary with both toxicity and bioavailability to aquatic biota. While there are standardized protocols to test the acute toxicity of sediment-borne compounds to aquatic invertebrates, there are none for assessing bioavailability to fish. We found that sediment-borne PAH caused an exposure-dependent induction of cytochrome P450 (CYP1A) enzymes in rainbow trout (Oncorhynchus mykiss) fingerlings exposed in semi-static 96 h bioassays, as shown by increased activity of ethoxyresorufin-o-deethylase (EROD). Assuming that PAH are taken up by trout due to partitioning from organic and inorganic constituents of sediments, we tested the effect of different test variables on bioaccumulation using induction as an index of exposure. EROD activity increased with exposure of fish to increasing volumes of sediments containing PAH, i.e., with increasing ratios of sediment to water. Uptake of single compounds from sandy sediments did not differ from uptake from clay or low organic (7% LOI — loss on ignition) sediments, but decreased when organic content was very high (58% LOI). Maximum induction was observed within 24~h of exposure and at 7.5^C relative to 15, 22, or 28^C. Storage and handling techniques had minor effects on sediment EROD induction potency. Absolute EROD activity was greater in white sucker (Catostomus commersoni) a benthic species, than in trout, a pelagic species. However, when basal (negative control) activity was accounted for, there was no difference in response between the species. Together, these experiments provide a basis for developing a standard protocol to test the bioavailability to fish of sediment-borne PAH.     

城市垃圾填埋场渗滤液中污染物检测与分析

垃圾渗滤液水质复杂,危害性大,对其无害化处理尚未得到很好解决。溶解性有机物(DOM)是导致渗滤液处理难以达标的主导性污染物。采用GC-MS技术可将渗滤液中DOM标识划分为亲水性和疏水性两类物质,为DOM的针对性处理提供了新的思路。采用ICP-AES从渗滤液中检测出美国环境保护局颁布的13种优先污染重金属中的11种,其含量大幅超标。废弃的电子材料、颜料涂料、电池以及药物等物质的不分类收集是导致渗滤液中重金属浓度超标、毒性增强的主要原因。在垃圾渗滤液处理领域,认为BOD5/COD小于0.3就不可以采用生物法处理的传统观点已经不符合实验与工程现实。通过检测与分析,掌握了垃圾渗滤液的水质特性与污染源头,能够为制定切实可行的处理方案提供借鉴。     

Polybrominated diphenyl ethers (PBDEs) and 2,2′,4,4′,5,5′-hexabromobiphenyl (BB-153) in landfill leachate in Cape Town, South Africa

An assessment of the concentrations of selected polybrominated diphenyl ether (PBDE) congeners as well as BB 153 in leachate samples collected from three landfill sites within the city of Cape Town was conducted. A liquid–liquid extraction technique was employed for the isolation of all the target compounds from the leachate samples. Extracts obtained were further subjected to multi-layer column chromatography employing different forms of silica gel. The prepared samples were analysed using a high capillary gas chromatograph equipped with a micro-electron capture detector (GC-μECD). The overall mean concentrations of the total PBDEs, including BDE 209 ranged between 5.65 and 2,240, 0.28–20.5 and 1.66–1,170 ng/l for Bellville, Coastal Park, and Vissershok landfill sites, respectively. The mean concentrations of BB 153, which were generally low in most of the samples analysed, were 70.4, 7.14 and 8.16 ng/l for Bellville, Coastal Park and Vissershok sites, respectively. The influence of precipitation on the characteristics and quantity of leachate produced from the landfill sites investigated was most pronounced during the August/September sampling regime. Generally, the trend observed in this study clearly indicated a wide variation in the levels of these contaminants in all the landfill sites studied from one sampling period to the other. However, the principal component analysis revealed that the release of these contaminants might be associated with two or three possible sources. This study further confirmed the relevance of landfill leachate as an important source of PBDE contamination of the environment, especially the groundwater and surface water sources.     

Assessment of the impact of landfill on groundwater quality: A case study of the Pirana site in western India

In present study focus has been given on estimating quality and toxicity of waste with respect to heavy metals and its impact on groundwater quality, using statistical and empirical relationships between different hydrochemical data, so that easy monitoring may be possible which in turn help the sustainable management of landfill site and municipal solid waste. Samples of solid waste, leachate and groundwater were analyzed to evaluate the impact of leachates on groundwater through the comparison of their hydrochemical nature. Results suggest the existence of an empirical relationship between some specific indicator parameters like heavy metals of all three above mentioned sample type. Further, K/Mg ratio also indicates three groundwater samples heavily impacted from leachate contamination. A good number of samples are also showing higher values for and Pb than that of World Health Organization (WHO) drinking water regulation. Predominance of Fe and Zn in both groundwater and solid waste samples may be due to metal plating industries in the area. Factor analysis is used as a tool to explain observed relation between numerous variables in term of simpler relation, which may help to deduce the strength of relation. Positive loading of most of the factors for heavy metal clearly shows landfill impact on ground water quality especially along the hydraulic gradient. Cluster analysis, further substantiates the impact of landfill. Two major groups of samples obtained from cluster analysis suggest that one group comprises samples that are severely under the influence of landfill and contaminated leachates along the groundwater flow direction while other assorted with samples without having such influence.     

Assessing water quality in a tropical lake using biomarkers in zebrafish embryos: developmental toxicity and stress protein responses

In order to achieve a more substantial appraisal of lake water quality, the assessment must not be based only on chemical measurements and analyses of the water itself, but even more so on the impact of existing conditions on aquatic biota. This is possible with the use of biotests or biomarkers, e.g. investigations of the developmental parameters (96-h embryotoxicity evaluate) or of the induction of heat shock proteins (proteotoxicity evaluate). To evaluate the suitability of these tests for environmental screening, fertilized zebrafish eggs were exposed to water samples collected from five sites of varying levels of stress from Laguna Lake, Philippines. Reconstituted water was used as laboratory control while water samples from a highly polluted freshwater source was used as positive control. Developmental parameters were noted and described within 48 and 96 h of exposure. Dilution experiments of the positive control were also done to further assess and compare toxicity potentials of Laguna Lake waters with those originating from a polluted freshwater. After the 96-h exposure, the levels of stress proteins (hsp 70) were determined in embryos from all exposure groups. Results showed 100% mortality in embryos exposed to undiluted positive control (PC) within only 12 h. Increasing dilution levels, however, resulted in lower mortality and lower abnormality rates. No detectable developmental differences were noted among embryos exposed to either the laboratory control or Laguna Lake waters at the end of 96 h, regardless of the source. Very high survival rates and high hatching success rates were observed in embryos exposed to lake waters as well as laboratory control, and the data did not differ significantly among the groups. Likewise, no significant malformations were noted among all developing embryos throughout the exposure period. However, the levels of heat shock proteins in the two sites located closest to Manila, the Philippine capital (Northern West Bay and Central West Bay), showed a pronounced elevation relative to the control, indicating that these stress proteins protect the embryos from the detrimental effects of pollutants in the water. Based on the 96-h early life stage (ELS) test, the water quality of the lake is good for fishery propagation despite the current levels of pollutants in the water. This finding is in accordance with the Class C status (i.e. suitable for fish growth and propagation) as given to the lake by the local environmental agency. On the other hand, data on proteotoxicity showed that the fish are under stress, presumably deriving from pollutants. This calls for a continuous monitoring and improvement of the lake water. The present study indicates that the two biomarker methods are very easy to use, practical, rapid, and sensitive for assessing water quality in a tropical lake and recommends for their incorporation into the future monitoring program of Laguna Lake.     

Artemia salina as test organism for assessment of acute toxicity of leachate water from landfills

Artemia salina has, for the first time, been used as test organism for acute toxicity of leachate water from three landfills (the municipal landfills at Kristianstad, Sweden and Siauliai, Lithuania, and an industrial landfill at Stena fragmenting AB, Halmstad, as well as for leachate from Kristianstad treated in different ways in a pilot plan). Artemia can tolerate the high concentrations of chloride ions found in such waters. Large differences in toxicities were found, the leachate from Siauliai being the most toxic one. To increase the selectivity in the measurements, a fractionation was done by using ion exchange to separate ammonium/ammonia and metal ions from the leachate, and activated carbon adsorbents for organic pollutants. The influence of some metals and phenol compounds on the toxicity was investigated separately. It was found that most of the toxicity emanated from the ammonium/ammonia components in the leachate. However, there was also a significant contribution from organic pollutants, other than phenol compounds, since separate experiments had in this latter case indicated negligible impact. The concentrations of metals were at a level, shown by separate experiments, where only small contribution to the toxicity could be expected.     

The effectiveness of composite lining systems in controlling the leakage of leachate from sanitary landfills to groundwater

Leachate, the hazardous liquid that percolated through the refuse layers of a sanitary landfill, if it leaks through the landfill lining system, can become a serious source of groundwater pollution. In the past, leaks have been detected in many landfills lined with flexible membrane liners (FML) whose failure may be attributed to flaws such as imperfect seaming, rips, and tears of the membrane, or from chemical attack that dissolves the membrane. Recent studies have shown that composite lining systems which include either a clayey subbase or a layer of geotextile in addition to the FML, can substantially reduce the leakage of leachate. Therefore in this study, four different lining systems are proposed and evaluated to determine their effectiveness in controlling leachate flow under various degree of flaws (referred to as leakage fraction LF) in the FML. The Hydrologic Evaluation of Landfill Performance (HELP) computer model of the Environmental Protection Agency of USA, currently the most widely accepted model for predicting the performance of leachate collection systems in that country, is used to evaluate the following lining systems: (1) a single FML or liner, (2) a single FML with a clayey composite, (3) a single FML with a geotextile called Claymax, and (4) a double FML. Based on the climatic conditions and the present lining construction cost of Alaska, the study shows that a single FML or liner is the most economical but it is also the least effective in controlling leachate flow. Design (3), a single FML with a geotextile, costs about 50 percent more but it reduces the leakage of leachate by several orders. Design (2) is also effective but the cost incurred in constructing a 3 feet thick clayey subbase is prohibitive and thus to effectively and economically minimize the hazards of potential groundwater contamination by leachate, Design (3) is recommended as the composite lining system for future landfill sites.     

Measurement of biochemical oxygen demand of the leachates

Biochemical oxygen demand (BOD) of the leachates originally from the different types of landfill sites was studied based on the data measured using the two manometric methods. The measurements of BOD using the dilution method were carried out to assess the typical physicochemical and biological characteristics of the leachates together with some other parameters. The linear regression analysis was used to predict rate constants for biochemical reactions and ultimate BOD values of the different leachates. The rate of a biochemical reaction implicated in microbial biodegradation of pollutants depends on the leachate characteristics, mass of contaminant in the leachate, and nature of the leachate. Character of leachate samples for BOD analysis of using the different methods may differ significantly during the experimental period, resulting in different BOD values. This work intends to verify effect of the different dilutions for the manometric method tests on the BOD concentrations of the leachate samples to contribute to the assessment of reaction rate and microbial consumption of oxygen.     

Effects of fulvic substances on the distribution and migration of Hg in landfill leachate

Mercury (Hg) distribution and migration in different landfill stabilization processes were evaluated in this study. Wide ranges of Hg concentrations were observed because of the heterogeneity and variability of landfill refuse. In addition, temporally variable conditions, including pH, organic matter, and vegetation cover, which influence Hg migration in landfills, may also affect the temporal distribution of Hg in landfill refuse. The main fraction of Hg, elemental Hg, decreased with time, while the stable fractions of Hg increased. The fulvic acid (FA) extracted from the landfill leachate had much lower overall Hg-complexation stability constants, which suggests that organic S groups might have been rapidly saturated by small amounts of Hg while leaving oxygen functional groups, such as carboxylic functional or phenolic groups, acting as the primary binding sites for Hg.     

刚果红-巯基棉分离富集光度法测定垃圾渗滤液中镉

基于在pH值为4.0～5.2的BR缓冲介质中,镉与刚果红发生显色反应并生成蓝色离子缔合物的原理,建立了测定垃圾渗滤液中镉的巯基棉分离富集光度法。其最大正吸收波长和最大负吸收波长分别为604nm和492nm,线性范围分别为0.10mg／L～2.80mg／L和0.10mg／L～3．40mg／L,采用双波长叠加测定灵敏度更高。探讨了渗滤液的处理方法及共存离子对测定的影响,优化了试验条件,实际样品的测定结果与原子吸收法对比无显著差异,加标回收率为98,6％～104％。     

The use of isotopes to identify landfill gas effects on groundwater

An evaluation of the source of volatile organic compounds in groundwater samples was performed at a landfill in southern California. The 3H (tritium) content of the water in leachate and water from the gas-collection system (condensed water and entrained water droplets) and the delta 13C and 14C content of the inorganic carbon in landfill gas CO2, leachate, and gas-collection system water were used to characterize the dissolved inorganic carbon (DIC) inside the landfill, while the same parameters were monitored in groundwater samples from affected monitoring wells and an unaffected well. Tritium levels from leachate and gas-collection system condensate ranged from approximately 2000 TU to over 4000 TU, orders of magnitude higher than unaffected groundwater. The average 14C content of DIC in the landfill pore-water samples was 121 pMC and the 14C content of unaffected groundwater DIC was 93 pMC, while the 14C content of the dissolved inorganic carbon in groundwater with VOC detections ranged from 105 to 119 pMC. The delta 13C of DIC in pore water was consistently above 0 per thousand and the delta 13C of unaffected groundwater DIC was -20.3 per thousand, while the delta 13C of DIC in affected groundwater samples was increased from -17.3 to -13.2 per thousand. The increases in both delta 13C and 14C in landfill gas-impacted groundwater DIC generally correlated with the number of volatile organic compounds detected and their concentrations. Based on the tritium and DIC 14C levels in leachate and water from the gas-collection system compared to those of unaffected water, significant increases in the tritium content of the water would be expected to accompany VOC detections and increases in delta 13C and 14C caused by landfill water. The results rule out landfill water as the VOC source, leaving landfill gas as the source. The identities and concentrations of the specific VOCs in affected groundwater samples varied among wells as well as between two leachate samples, ruling out the use of a VOC "fingerprint" for leachate or landfill gas to be compared to groundwater VOC concentrations.     

毒性废水稀释中强度改变与浓度改变的跟随性研究

适用于已知毒物和毒性应用剂量测定的经典技术方法,以样品稀释过程中毒性强度改变与浓度改变的等比率跟随性为基础。废水中的毒性物质常以未知和混合为特征,高毒性的样品现状浓度距离方法预试验中最小全致死浓度较远,以已知毒物重铬酸钾毒性为对照,形成5种毒物高层级等强度毒性,通过这些高毒性废水同步等比率稀释后毒性改变的同步性测定试验,证明毒性废水稀释中强度改变与浓度改变的跟随状态具有不确定性。     

Induction of cytochrome P4501A and endocrine disrupting effects of school incinerator residues

The emission of the dioxin-like compounds from on-site waste incinerators of seven schools in Kyonggi Province of Korea was evaluated by determination of the cytochrome P4501A(CYP1A) catalytic activity and antiestrogenic activity using cell culturemicrobioassay. The residue samples were extracted in a Soxhlet apparatus using toluene for 20 hr. The concentrated crude extractswere fractionated with a basic alumina column. Dioxin-like compounds were then extracted. Induction of CYP1A activity in a rat(H4IIE) hepatoma cell line was used as indicator of biologicaleffect of incinerator residues and measured as 7-ethoxyresorufin-O-deethylase(EROD) activities. The EROD activities of fraction II extracts (one of the two extracts) in the H4IIE cells were from 0.044±0.002 to 4.424±0.351 ng-TEQ g^-1 (TCDD Toxicity equivalent), showing relatively high inducing capacity. Antisetrogenicity of the extracts was measured as decrease in E₂-induced cell proliferation. Most of the extracts showed antiestrogenic activity in MCF7-BUS cell.The TEQ levels of the incinerator residues and the antiestrogenicactivities were in good correlation, strongly suggesting that thepotent toxic emissions were indeed produced from the on-site school waste semi-incinerators and could cause the antiestrogenicity.     

Performance of bioreactor landfill with waste mined from a dumpsite

Emissions from landfills via leachate and gas are influenced by state and stability of the organic matter in the solid waste and the environmental conditions within the landfill. This paper describes a modified, ecologically sound waste treatment technique, where municipal solid waste is anaerobically treated in a lysimeter-scale landfill bioreactor with leachate recirculation to enhance organic degradation. The results demonstrate a substantial decrease in organic matter (BOD 99%, COD 88% and TOC 81%) and a clear decrease in nutrient concentrations especially ammonia (85%) over a period of 1 year with leachate recirculation.     

水体中黑碳对斑马鱼胚胎的生长发育危害识别

黑碳(BC)是含碳物质不完全燃烧后产生的残留物,广泛分布于水体、土壤和大气中,目前对于BC对水生生物的生态效应研究还十分有限。通过燃烧煤油获得BC,以斑马鱼胚胎为模式生物,研究了质量浓度分别为0,0.05,0.5和5 mg/L的BC悬浮溶液对斑马鱼早期生命阶段的生长发育毒性。结果表明,煤油源BC对斑马鱼具有明显的毒性作用,与对照组相比,BC暴露明显增加了斑马鱼的死亡率和心率,显著降低了孵化率和体长,导致斑马鱼形态畸形并且畸形率呈浓度依赖性。5mg/L的BC对斑马鱼的各项生长发育指标影响并非是最大的,可能与BC颗粒在水溶液中的聚集有关。该研究为BC在水生环境中的危害识别提供实验依据。     

Monitoring Pollution in River MureŞ, Romania, Part III: biochemical effect markers in fish and integrative reflection

Along a downstream stretch of River Mureş, Romania, adult males of two feral fish species, European chub (Leuciscus cephalus) and sneep (Chondrostoma nasus) were sampled at four sites with different levels of contamination. Fish were analysed for the biochemical markers hsp70 (in liver and gills) and hepatic EROD activity, as well as several biometrical parameters (age, length, wet weight, condition factor). None of the biochemical markers correlated with any biometrical parameter, thus biomarker reactions were related to site-specific criteria. While the hepatic hsp70 level did not differ among the sites, significant elevation of the hsp70 level in the gills revealed proteotoxic damage in chub at the most upstream site, where we recorded the highest heavy metal contamination of the investigated stretch, and in both chub and sneep at the site right downstream of the city of Arad. In both species, significantly elevated hepatic EROD activity downstream of Arad indicated that fish from these sites are also exposed to organic chemicals. The results were indicative of impaired fish health at least at three of the four investigated sites. The approach to relate biomarker responses to analytical data on pollution was shown to fit well the recent EU demands on further enhanced efforts in the monitoring of Romanian water quality.     

Evaluation of analytical strategies for the determination of metal concentrations to assess landfill leachate contamination

Due to the complex nature of landfill leachates, metal and metalloid analyses prove to be tricky and suffer from a lack of standard protocols. A complete approach has been adopted to investigate the influence of the different steps during the sample processing of French landfill leachates. The validation of the entire protocol has been achieved using a laboratory reference material. This material, which is a real landfill leachate, is representative of real samples. Its evaluation has allowed a quality control for metal and metalloid analyses in landfill leachates. Precautions concerning storage temperature, aeration and filtration are proposed to perform accurate metal analyses in these complex matrices. The sample processing has been applied to the seasonal monitoring of a French landfill. The assessment of major leachate metallic contaminants such as As, Cr, Sb, Sn, has been performed by evaluating the relative enrichment of metals and metalloids in comparison with rain water and groundwater. In addition, hydrological data are useful and complementary information for pointing out the main factors affecting metal concentrations and thus their potential remobilisation pathways.     

徐州市生活垃圾填埋场地下水典型金属污染物研究

在对徐州地区生活垃圾填埋场调查分析的基础上,根据填埋特征和地质状况选取4座典型填埋场为调查对象,采用ICP-MS对垃圾渗滤液及地下水中26种重金属进行监测分析。结果表明,在4个垃圾填埋场地下水及渗滤液中ρ(Sr)均相对较高(700μg/L);地下水中金属元素均正在以Ca,Mg为主向Na,Mg为主转化;地下水中Al,B质量浓度大小顺序为Y场(雁群)S场(睢宁)C场(翠屏山)P场(邳州);4个垃圾填埋场共同典型重金属污染物为Mn,Fe,Zn,Ba;除上述污染物外,Y场地下水潜在典型重金属污染物为Pb和Mo;S场为Mo和As;C场为Tl和Co;P场为As。