Chronic toxicity of the Sava River (SE Europe) sediments and river water to the algae Pseudokirchneriella subcapitata

The Sava River, the largest and most commercially valuable water body in the riparian countries, receives inputs of organic and inorganic compounds from a variety of domestic and industrial activities that may affect the health of human beings and wildlife. In this work, the chronic toxicity of sediment, sediment porewater and surface water from the Sava River and connecting tributaries to the freshwater algae Pseudokirchneriella subcapitata was assessed to characterise the potential impact on aquatic organisms. Samples of different environmental matrices were either tested directly (porewater) or subjected to pre-concentration (sediments and surface water) prior to testing in a 72 h growth inhibition assay using P. subcapitata. The results show that a combination of pre-concentration and bioassay testing was able to characterise the toxic potential and to identify several compartment-specific "hot spots" along the Sava River. Based on the resulting data, a crude risk assessment identified that some of the locations may represent a risk to algae. Additional testing using multiple species and exposure phases is required to conduct a thorough risk assessment, however.     

Experiments using Daphnia to measure landfill leachate toxicity

Leachate from municipal refuse was analysed for acute toxicity using Daphnia pulex, rainbow trout (Salmo gairdneri) and sockeye salmon (Oncorhynchus nerka) as test organisms. Procedures used in conducting the daphnia bioassay are detailed. The $\text{3}\text{1}\text{2}\text{-}\text{year}$ testing program was designed to compare daphnia bioassays with standard fish bioassays and residual oxygen bioassays (ROB). The results from the daphnia bioassays compared favourably with the standard fish bioassay results. No disadvantages could be seen in using daphnia rather than fish as test organisms for determining the acute toxicity of municipal refuse leachates. However, both financial and logistic advantages could be gained by using daphnia rather than fish.     

双效工程菌Y1溶藻产物的急性毒性与健康风险评估

为检测溶藻、藻毒素降解双效工程菌Y1溶藻产物的生物毒性,采用发光细菌测定急性毒性,利用USEPA推荐模型对水源水进行健康风险评估。结果表明,Y1溶藻菌液能有效地抑制铜绿微囊藻的生长,并对发光细菌具有轻微的毒性。当叶绿素a浓度为34.92mg/m~3时,增加投加量会使藻液毒性加强;当叶绿素a浓度大于46.56mg/m~3时,加菌后藻液毒性明显降低。水源水中MC-LR的非致癌风险值为2.89~4.87,通过BAF处理后能减少到1.3,而加Y1菌强化后得到的0.6达到了健康风险评估安全标准。UV254与致突变强度的预测模型表明经过处理的水中有机物无致突变性,说明Y1应用于水源水无生物毒性和致突变风险。     

Toxicity characterization of urban stormwater with bioanalytical tools

Stormwater harvesting has become an attractive alternative strategy to address the rising demand for urban water supply due to limited water sources and population growth. Nevertheless, urban stormwater is also a major source of surface water pollution. Runoff from different urban catchments with source contributions from anthropogenic activities and various land uses causes variable contaminant profiles, thus posing a challenging task for environmental monitoring and risk assessment. A thorough understanding of raw stormwater quality is essential to develop appropriate treatment facilities for potential indirect potable reuse of stormwater. While some of the key chemical components have previously been characterized, only scarce data are available on stormwater toxicity. We benchmarked stormwater samples from urban, residential and industrial sites across various Australian capital cities against samples from the entire water cycle, from sewage to drinking water. Six biological endpoints, targeting groups of chemicals with modes of toxic action of particular relevance for human and environmental health, were investigated: non-specific toxicity (Microtox and combined algae test), the specific modes of action of phytotoxicity (combined algae test), dioxin-like activity (AhR-CAFLUX), and estrogenicity (E-SCREEN), as well as reactive toxicity encompassing genotoxicity (umuC) and oxidative stress (AREc32). Non-specific toxicity was highly variable across sites. The baseline toxicity equivalent concentrations of the most polluted samples were similar to secondary treated effluent from wastewater treatment plants. Phytotoxicity results correlated well with the measured herbicide concentrations at all sites. High estrogenicity was found in two sampling events and could be related to sewage overflow. Genotoxicity, dioxin-like activity, and oxidative stress response were evident in only three of the samples where the stormwater drain was beside a heavy traffic road, confirming that road runoff is the potential source of contaminants, while the bioanalytical equivalent concentrations (BEQ) of these samples were similar to those of raw sewage. This study demonstrates the benefit of bioanalytical tools for screening-level stormwater quality assessment, forming the basis for the evaluation of future stormwater treatment and reuse schemes.     

Plasma vitellogenin levels and gonadal morphology of wild carp (Cyprinus carpio L.) in a receiving rivers downstream of sewage treatment plants

A field study was carried out in order to assess, in a wild cyprinid fish, the biological effects related to the exposure to estrogenic compounds. The gonadosomic index (GSI), plasma vitellogenin (VTG) level and histological observations of gonads were performed to evaluate the reproductive health in common carp (Cyprinus carpio) from three rivers receiving sewage treatment plant (STP) effluents (Guadarrama, Henares and Jarama rivers, Spain) and from a reference site (Lozoya river). No significative differences in male GSI between sites were described. Only in the 18% of fish from Guadarrama river were elevated plasma VTG levels detected. The presence of oocytes in the testis was not found in any of males examined. The results suggest that the released estrogenic compounds by STPs effluents are not enough capable to adversely affect the gonadal development of the fish studied. Additional studies will be required to assess the risk of estrogenic compounds on fish from other sites of this area.     

Toxic and genotoxic evaluation of six antibiotics on non-target organisms

The ecotoxicity of the following six antibiotics on aquatic organisms was investigated: Erythromycin, Oxytetracyclin, Sulfamethoxazole, Ofloxacin, Lincomycin and Clarithromycin. Bioassays were performed on bacteria, algae, rotifers, microcrustaceans and fish to assess acute and chronic toxicity, while SOS Chromotest and Ames test were used to detect the genotoxic potential of the investigated drugs. For risk assessment, the environmental impact was calculated by MEC/PNEC ratio using the available data from the literature regarding their occurrence in the aquatic environment and the toxicity data obtained from the bioassays performed. The ecotoxicological results showed that acute toxicity was in the order of mg/L while, for the chronic data the antibiotics were bioactive at concentrations in the order of microg/L, mainly for the algae. Drugs investigated were one or two order of magnitude less active against rotifers and crustaceans. Ofloxacin was the only genotoxic compound and Sulfamethoxazole, Ofloxacin and Lincomycin were mutagenic. As for environmental risk, the macrolides were found to be the most harmful for the aquatic environment.     

Multiscale approach of fish responses to different types of environmental contaminations: a case study

In order to study the responses of wild fish to environmental contaminations, different sites (two references and three contaminated) were sampled across the Walloon hydrographical network (southern Belgium). The status of fish communities was characterized according to an index of biotic integrity (IBI). Furthermore, population structure, reproductive parameters and biochemical assays were performed on chub (Leuciscus cephalus) as sentinel species. The results showed impaired responses in fish sampled downstream paper mill effluents (BKME), in a polychlorinated biphenyls (PCB) contaminated site, and in a river receiving pesticides (i.e. atrazine and hexachlorocyclohexane). High level of hepatic 7-ethoxyresorufin-Odeethylase (EROD) activity were evidence of direct toxicant exposure, while population structures showed unbalanced size-frequency distributions. Moreover, in the pesticide contaminated site, a higher proportion of atretic oocytes was found in females and abnormally high plasma alkali-labile phosphorus (ALP) values were noticed in males, compared to reference sites, suggesting that chubs were exposed to estrogenic compounds. IBI scores from both sites were also lower than in reference sites. On the other hand, no particular response was found in hepatic glutathione-S-transferase (GST) activities, hepatic heat-shock protein HSP70 concentrations and plasma sex steroids (estradiol-17beta, 11-ketotestosterone and testosterone) in chubs from any sites. By using a multiparametric approach, a better understanding of response mechanisms to pollution in fish may be achieved in order to improve the ecological status of river ecosystems.     

A Daphnia magna feeding bioassay as a cost effective and ecological relevant sublethal toxicity test for Environmental Risk Assessment of toxic effluents

Environmental Risk Assessment of chemical products and effluents within EC countries require the use of cost effective standardized toxicity tests that in most cases are restricted to acute responses to high doses. Thus, subtle ecological effects are underestimated. Here we propose a short-term one day Daphnia magna feeding inhibition test as a cost effective and ecological relevant sublethal bioassay. The sensitivity and reliability of the proposed bioassay was tested in the laboratory against standardized bacteria, algae growth, D. magna and fish acute toxicity test by using 16 chemical mixture x water type combinations that included four different water types fortified with four complex chemical mixtures. Water types included ASTM hard water and three selected effluents diluted 1/10 in water to mimic worse field situations that many overexploited arid river ecosystems suffer during summer months when effluents are discharged into them with little dilution. The results obtained denoted a greater sensitivity of the proposed feeding bioassay in 51 out of 65 tests performed with an average sensitivity 50 fold greater than that of the standardized tests. The greater differences were obtained for mixtures that included narcotic chemicals and the lowest differences for those containing pesticides. Furthermore, feeding responses to the studied contaminant mixtures behaved differently to increasing TOC content than those based on bioluminescent bacteria and algae. Increasing TOC coming from sewage treated effluents decrease toxicity to the latter bioassays but increased those of D. magna feeding bioassays. These results empathize the need to include additional bioassays to monitor more accurately and realistically the toxicity of effluents or surface waters dominated by effluent discharges, a quite common situation in America and Mediterranean arid regions.     

Selecting analytical target pesticides in monitoring: Sensitivity analysis and scoring

Measuring river water concentrations of all pesticides applied in a catchment area is a daunting task. This study aims to develop new score tables for selecting analytical target pesticides. Sensitivity analyses were conducted using a diffuse pollution hydrologic model to quantitatively evaluate the influence of pesticide properties (e.g., log K_OC, degradability [half-life]) on concentrations of rice-farming pesticides in river water. Using the results of the analyses, score tables were systematically designed for the pesticide properties such that the sum of the scores for a particular pesticide, designated as the contamination index, was proportional to the expected/predicted concentration of that pesticide in river water. The contamination indexes for pesticides applied in three river basins were calculated and compared with the corresponding observed pesticide concentrations. Correlations between contamination indexes and observed concentrations were fairly good. Pesticides were ranked according to the quotients obtained by dividing the pesticide concentrations predicted from the contamination indexes by the corresponding drinking-water quality guideline values, and pesticide candidates to be monitored were successfully selected on the basis of a threshold quotient.     

A video-based movement analysis system to quantify behavioral stress responses of fish

Behavioral alterations can be measured as endpoints for sublethal toxicity, and serve as a tool for environmental risk assessment and analysis of toxicological impact. Numerous technical and biological factors have made sublethal effects on fish behavior difficult to quantify. In order to investigate stress- and contaminant-induced behavioral alterations, a video analysis system was designed by our laboratory. With this system up to 12 fish may be individually housed in 20 L exposure arenas and automatically videotaped at multiple and discrete intervals during an experimental period. Analog video data can then digitized, converted into x,y coordinates, and finally transformed into relevant behavioral endpoints using software designed for tracking fish movement combined with specific algorithms. These endpoints include velocity, total distance traveled, angular change, percent movement, space utilization, and fractal dimension (path complexity). Data from fish exposed to a reference toxicant, MS222, and simulation experiments, are presented to exemplify alterations in fish behavior associated with exposure, and accuracy and precision, respectively. The system provides flexibility to analyze any observed movement behavior, is remotely controlled, and can be transportable. These movement analyses can be used to identify characteristic behavioral responses to a variety of environmentally-relevant stressors, and assist in risk assessment and the development of more sensitive lowest observable effect level and no observable effect level for sentinel species.     

Application of short-term bioassay guided chemical analysis for water quality of agricultural land run-off.

The effect of agricultural land run-off on the water quality of Lake Kojima, Japan, was investigated using a short-term bioassay-guided chemical analysis. Water samples were collected for 1 year starting from June 1995 to June 1996. Toxicity of the dissolved and adsorbed extracts in the water samples was evaluated using the Daphnia immobilization test and the concentrations of pesticides and putative toxic substance in the extracts were determined by high performance liquid chromatography. Most of the dissolved extracts caused immobilization of the test Daphnia magna at low concentrations during the period of paddy pesticide application. Some extracts were found to contain pesticides such as dymron, mefenacet and flutolanil, but their concentrations were too low to have a toxic effect on the daphnia. An unknown toxic compound, Peak C, was isolated from some river water samples, but it produced only a relatively weak toxicity to Daphnia. To better understand the impact of agricultural run-off on a receiving water body, the relationship between the observed toxicity and the concentrations of pesticides and Peak C in the water samples was studied both temporally and spatially.     

Comparison of self-organizing maps classification approach with cluster and principal components analysis for large environmental data sets

Three classification techniques (loading and score projections based on principal components analysis (PCA), cluster analysis (CA) and self-organizing maps (SOM)) were applied to a large environmental data set of chemical indicators of river water quality. The study was carried out by using long-term water quality monitoring data. The advantages of SOM algorithm and its classification and visualization ability for large environmental data sets are stressed. The results obtained allowed detecting natural clusters of monitoring locations with similar water quality type and identifying important discriminant variables responsible for the clustering. SOM clustering allows simultaneous observation of both spatial and temporal changes in water quality. The chemometric approach revealed different patterns of monitoring sites conditionally named "tributary", "urban", "rural" or "background". This objective separation could lead to an optimization of river monitoring nets and to a better tracing natural and anthropogenic changes along the river stream.     

Fluorescent whitening agents: Acute fish toxicity and accumulation studies

Four fluorescent whitening agents (FWA), three of which are currently marketed in the United States for use as ingredients in household laundry detergents, were tested to determine their acute toxicity to the bluegill (Lepomis macrochirus), as well as their potential for significant accumulation in the flesh of the bluegill and channel catfish (Ictalurus punctatus), two species of fish common to U.S. waters. Results of the acute toxicity bioassays showed that no acutely toxic effects on fish, caused by these materials, would be expected at concentrations well above their projected environmental levels. Rates of accumulation and maximum levels accumulated were evaluated in the laboratory for a period of 90–105 days, followed by an examination of the rate of elimination of these materials from the fish over a 28-day period. Under these conditions, neither species accumulated any of three anionic fluorescent whitening agents (sulfonated stilbene derivatives) when exposed to nominal concentrations of 0.125, 1.25, or 12.5 μg 1⁻¹ in water. A nonionic FWA not currently used in the U.S. detergent formulations was significantly accumulated at the two highest concentrations tested. The nonionic FWA accumulated was rapidly eliminated by fish upon their transfer to water devoid of this chemical. Elimination of FWA was essentially complete by 14 days. No significant accumulation by fish was detected with any FWA when exposure was conducted at levels approximating projected environmental concentrations. Details of the acute toxicity tests, and the patterns of accumulation and elimination seen in this study, as well as their significance, are discussed.     

Comparison of the toxicity thresholds of water pollutants to bacteria, algae, and protozoa in the cell multiplication inhibition test

Using analogous methods of the cell multiplication inhibition test, the toxicity threshold (TT) of 156 potential water pollutants was determined for model organisms of biological self-purification, i.e. bacteria of the species Pseudomonas putida, green algae of the species Scenedesmus quadricauda, and flagellate protozoa of the species Entosiphon sulcatum. From the number of 156 inorganic and organic pollutants tested, 23 exhibited a pronounced selective toxic action on bacteria of the species Pseudomonas putida, 47 on algae of the species Scenedesmus quadricauda, and 43 on flagellate protozoa of the species Entosiphon sulcatum. It is seen from the ratios found that a determination of the toxicity threshold of potential water pollutants for these three model organisms from the microbiological spectrum would provide a broader basis for assessing the damaging action of water pollutants to model organisms active in the biological self-purification of water. Ecotoxicological testing of potential water pollutants to evaluate their toxicity involving only one model organism of biological self-purification would give an incomplete and biased picture of the effects of pollutants.     

Runoff rates and ecotoxicity of zinc induced by atmospheric corrosion

Initiated by regulatory restrictions on the use of zinc for various building and construction applications, together with a lack of knowledge related to the release of zinc induced by atmospheric corrosion, a major interdisciplinary research project was implemented to generate data to be used in future risk assessment. Runoff rates from a large number of commercially available zinc-based materials have been determined on panels inclined 45 degrees from the horizon, facing south, during a 1-year atmospheric exposure in an urban environment in Sweden. Possible environmental effects of runoff water immediately after leaving the surface of the various materials have been evaluated during two different sampling periods of varying season and zinc concentration, using the standard growth inhibition test with algae. Raphidocelis subcapitata (formerly Selenastrum capricornutum). Zinc-specific biosensors with the bacterial strain of Alcaligenes eutrophus, and computer modeling using the water-ligand model MINTEQA2 and the humic aquatic model WHAM, have been used to assess the bioavailability and chemical speciation of zinc in the runoff water. An excellent consistency between the different methods was observed. The results show considerably lower runoff rates of zinc (0.07-3.5 g m(-2) year(-1)) than previously being used for regulatory restrictions, and the concentration of zinc to be predominantly responsible for the observed toxicity of the runoff water towards the green algae. The majority of the released zinc quantity was found to be present as free hydrated zinc ions and, hence, bioavailable. The data do not consider changes in bioavailability and chemical speciation or dilution effects during entry into the environment, and should therefore only be used as an initial assessment of the potential environmental effect of zinc runoff from building applications. This interdisciplinary approach has the potential for studies on the environmental fate of zinc in soil or aquatic systems.     

Use of highly sensitive sublethal stress responses in the social amoeba Dictyostelium discoideum for an assessment of freshwater quality

In this work, the sensitivity of a battery of tests on the social amoeba Dictyostelium discoideum has been assessed within a freshwater toxicity study. The results obtained from the evaluation of survival and replication rate of D. discoideum were compared to those derived with a series of widely used tests for freshwater toxicity assessment, i. e. bioassays using Vibrio fischeri, Daphnia magna and Pseudokirchneriella subcapitata. The effects on sublethal endpoints, i.e. lysosomal membrane stability (LMS) and endocytotic rate, were analysed in conjunction with high-level endpoints to verify the potential to make a typical bioassay more sensitive. The field ecotoxicological investigation employing D. discoideum is part of a monitoring study assessing environmental quality of the Bormida River (Italy), subjected until recently to a chronic industrial pollution. The survey was carried out at several stations (upstream and downstream of a chemical factory outlet) in two different periods. In 2002, the results of chemical analyses performed on river water indicated no contamination. The ecotoxicological data obtained in this period showed that no evidence of biological effects was observed using V. fischeri and D. magna bioassays. In spite of the previous classical acute toxicity tests, significant differences in cell viability of D. discoideum were found. By analysing the effects measured on LMS and endocytotic rate, more relevant changes were observed for these sublethal stress biomarkers compared to survival. The chronic toxicity data showed significant changes in cell growth both of P. subcapitata and D. discoideum. Nevertheless, more sensitive and rapid responses were obtained when assessing the effects of exposure on D. discoideum. The chemical and ecotoxicological data obtained in 2006 indicated a full recovery of the quality of the river water (neither contamination nor toxicity found). Altogether, the results reported in this study underline that the use of a battery of biomarkers in conjunction with high-level endpoints may help follow the pollutant-induced stress syndrome in the organisms from early sublethal effects to starting mortality.     

泾河平凉段水环境容量分析与对策

水环境容量是衡量一个水域水质状况的重要指标,泾河作为平凉市的主要河流,其环境状况与人们的生产、生活息息相关。将泾河平凉段分为三个控制单元,分别分析其属性和水质达标情况,在对污染源进行调查研究之后,采用一维模型计算其水环境容量,并将其与现行排污量进行比较,得出该流域的剩余水环境容量,同时提出了泾河平凉段水环境治理的具体对策。     

Mercury contamination in the vicinity of a derelict chlor-alkali plant Part II: contamination of the aquatic and terrestrial food chain and potential risks to the local population

This study investigated the environmental impact and level of risk associated with mercury (Hg) contamination near a derelict chlor-alkali plant in Pavlodar, Northern Kazakhstan. Several species of fish were sampled from the highly polluted Lake Balkyldak and the nearby river Irtysh, to assess the extent of Hg bioaccumulation in the aquatic food chain and potential human health risks. A small number of bovine tissue samples, water samples, soil and plant samples from a nearby village were also investigated in order to make a preliminary assessment of potential impacts on the terrestrial food chain. Mercury levels in fish caught from Lake Balkyldak ranged from 0.16 to 2.2 mg kg(-1) and the majority of fish exceeded current human health criteria for Hg. Interspecies comparisons indicated that Hg is accumulated in the order dace>carp>tench. Site-specific bioaccumulation factors (BAF) were calculated for THg, and were estimated for MeHg. Fish from the river Irtysh and floodplain oxbow lakes contained between 0.075 and 0.159 mg kg(-1) of Hg and can be regarded as uncontaminated. Soils were found to be impacted by past atmospheric emissions of Hg. Cattle grazing in the surroundings of the factory are exposed to Hg from contaminated soils, plants and surface water, but the consumption of contaminated fish from the lake appears to be the main route of exposure for humans.     

Genes and environment - striking the fine balance between sophisticated biomonitoring and true functional environmental genomics

This article provides an overview how the application of the gene profiling (mainly via microarray technology) can be used in different organisms to address issues of environmental importance. Only recently, environmental sciences, including ecotoxicology, and molecular biology have started to mutually fertilize each other. This conceptual blend has enabled the identification of the interaction between molecular events and whole animal and population responses. Likewise, striking the fine balance between biomonitoring and functional environmental genomics will allow legislative and administrative measures to be based on a more robust platform. The application of DNA microarrays to ecotoxicogenomics links ecotoxicological effects of exposure with expression profiles of several thousand genes. The gene expression profiles are altered during toxicity, as either a direct or indirect result of toxicant exposure and the comparison of numerous specific expression profiles facilitates the differentiation between intoxication and true responses to environmental stressors. Furthermore, the application of microarrays provides the means to identify complex pathways and strategies that an exposed organism applies in response to environmental stressors. This review will present evidence that the widespread phenomenon of hormesis has a genetic basis that goes beyond an adaptive response. Some more practical advantages emerge: the toxicological assessment of complex mixtures, such as effluents or sediments, as well as drugs seems feasible, especially when classical ecotoxicological tests have failed. The review of available information demonstrates the advantages of microarray application to environmental issues spanning from bacteria, over algae and spermatophytes, to invertebrates (nematode Caenorhabditis elegans, crustacea Daphnia spp., earthworms), and various fish species. Microarrays have also highlighted why populations of a given species respond differently to similar contaminations. Furthermore, this review points at inherent limits of microarrays which may not yet have been properly addressed, namely epigenetics, which may explain heritable variation observed in natural population that cannot be explained by differences in the DNA sequence. Finally, the review will address promising future molecular biological developments which may supersede the microarray technique.     

Impact of calcium and TOC on biological acidification assessment in Norwegian rivers

Acidification continues to be a major impact in freshwaters of northern Europe, and the biotic response to chemical recovery from acidification is often not a straightforward process. The focus on biological recovery is relevant within the context of the EU Water Framework Directive, where a biological monitoring system is needed that detects differences in fauna and flora compared to undisturbed reference conditions. In order to verify true reference sites for biological analyses, expected river pH is modeled based on Ca and TOC, and 94% of variability in pH at reference sites is explained by Ca alone, while 98% is explained by a combination of Ca and TOC. Based on 59 samples from 28 reference sites, compared to 547 samples from 285 non-reference sites, the impact of calcium and total organic carbon (TOC) on benthic algae species composition, expressed as acidification index periphyton (AIP), is analyzed. Rivers with a high Ca concentration have a naturally higher AIP, and TOC affects reference AIP only at low Ca concentrations. Four biological river types are needed for assessment of river acidification in Norway based on benthic algae: very calcium-poor, humic rivers (Ca < 1 mg/l and TOC > 2 mg/l); very calcium-poor, clear rivers (Ca < 1 mg/l and TOC < 2 mg/l); calcium-poor rivers (Ca between 1 and 4 mg/l); moderately calcium rich rivers (Ca > 4 mg/l). A biological assessment system for river acidification in Norway based on benthic algae is presented, following the demands of the Water Framework Directive.