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.     

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.