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Gammarids are aquatic amphipods widely used for water quality monitoring. To investigate the copper and cadmium diet-borne metal uptake in Gammarus pulex, we adapted the pulse-chase stable isotopes-based approach to determine the food ingestion rate (IR), the gut retention time (GRT) and the metal assimilation efficiencies (AE). G. pulex were fed with (65)Cu-, (106)Cd-, and (53)Cr-labeled alder leaves for 7.5h and then with unlabeled leaves for 5d. The metal stable isotope contents in the gammarids, leaves, filtered water and periodically collected feces were determined. Chromium was poorly assimilated by the gammarids; thus, Cr was used as an unassimilated tracer. The first tracer defecation occurred before the first feces harvest, indicating a gut passage time of less than 9h. A 24-h GRT and a 0.69gg(-1)d(-1) IR were estimated. The Cd AE value was estimated as 5-47%, depending on the assimilation determination method applied. The Cu AE value could not be evaluated regardless of the determination method used, most likely because of the rapid Cu regulation in gammarids in addition to analytical uncertainties when determining the Cu content in leaves. Application of the Cd AE value in the framework of the biodynamic bioaccumulation model shows that the diet-borne uptake of Cd significantly contributes (66-95%) to the metal bioaccumulation in G. pulex fed with alder leaves., (Copyright © 2014 Elsevier Inc. All rights reserved.)

It is difficult to assess the biological consequences of diffuse water contamination by micropollutants which are present in rivers at low, even sublethal levels. River biofilms, which respond quickly to changes of environmental parameters, are good candidates to acquire knowledge on the response of aquatic organisms to diffuse chemical contamination in the field. The study was designed as an attempt to link biofilm metal tolerance and metallic contamination in a field survey covering 13 different sampling sites in the Seine river basin (north of France) with low contamination levels. Cd and Zn tolerance of heterotrophic communities was assessed using a short-term toxicity test based on β-glucosidase activity. Metal tolerance levels varied between sites but there was no obvious correlation between tolerance and corresponding water contamination levels for Cd and Zn. Indeed, metallic contamination at the sampling sites remained subtle when compared to water quality standards (only two sampling sites had either Zn or both Cu and Zn concentrations exceeding the Environmental Quality Standards set by the EU Water Framework Directive). Yet, multivariate analysis of the data using Partial Least Squares Regression revealed that both metallic and environmental parameters were important variables explaining the variability of metal tolerance levels. Automated Ribosomal Intergenic Spacer Analysis (ARISA) was also performed on both bacterial and eukaryotic biofilm communities from the 13 sampling sites. Multivariate analysis of ARISA fingerprints revealed that biofilms with similar tolerance levels have similar ARISA profiles. Those results confirm that river biofilms are potential indicators of low, diffuse contamination levels of aquatic systems., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Biomonitoring, assumed to be an integrative measurement of the chemical exposure of aquatic organisms, is not straightforward for essential metals because they can be actively regulated by animals. Although increasing bioaccumulation with exposure levels is a crucial endpoint for the development of biomonitors, it is rarely verified in real environments, where the metal concentrations are rather low and vary little. This study was designed at the scale of a river basin to assess the ability of Gammarus pulex indigenous populations to accumulate Cu, Zn and Mn in realistic exposure conditions. During two annual campaigns, water and gammarids were collected at various sites contrasted in terms of physicochemistry and contamination. The results show significant relationships between metal concentrations in animals and in freshwaters established by conceptual models of bioaccumulation, but with patterns specific to each metal (base level, internal regulation and maximal accumulation). In particular, a saturation process of Cu accumulation occurs at environmental exposure levels, unlike Mn and Zn. Statistical analyses performed from field data show that Cu and Zn bioaccumulations may be influenced by a complex combination of geochemical variables, unlike Mn. We conclude that G. pulex is a useful candidate to monitor metal bioavailability in freshwaters due to its responsiveness to low exposures of surrounding environments. Nevertheless, a reliable quantification of bioavailability of essential metals requires characterizing some geochemical effects on metal bioaccumulation., (© 2013 Elsevier B.V. All rights reserved.)

Recent papers have alerted the scientific community that a diffusive boundary layer (DBL) forming in front of diffusive gradients in thin film (DGT) devices when they are immersed in water might have a significant impact on the results and have suggested a method to assess the DBL. This paper aims at evaluating to what extent the DBL impacts the results of metal measurement in water by DGT and providing new information on the dissociation kinetics of metal complexes in wastewater by using DBL calculation. A careful study of the influence of the water velocity on the measurement with DGTs equipped with restricted gels is presented. Deployments took place in the laboratory with a range of stirring speeds (0-400 rpm) and in a canal receiving treated wastewater with increasing controlled water velocity (0.07-3 cm s(-1)). Even under extreme low flow conditions, the error made in using the equation that does not take into account that the DBL was lower than the analytical error. Nevertheless, the DBL is the seat of dissociation of complexes and increases the lability window beyond the steric constraints of the hydrogel. The capacity of restricted gels to only sample inorganic species under these conditions is questioned. This study also is an opportunity to provide information on metal-ligand interactions in wastewater by creating the kinetic signature of the wastewater. Unlike previous studies which used different types of water, Pb was the more limited metal and interacted strongly with the ligands.

The present report is the companion study of our previous study in which we investigated the impact of the dissolved organic matter, water cationic composition and pH on the bioavailability and the bioaccumulation of copper (Cu) in aquatic mosses (Fontinalis antipyretica). The impact had been assessed under laboratory controlled conditions and modelled using a two-compartment model calibrated under a wide range of water compositions (Ferreira et al., 2008, 2009). Herein are reported the validation stage of the abovementioned approach for contrasted geochemical field conditions. Experiments were performed with aquatic mosses that were exposed for 7d to two nominal Cu concentrations (5 and 15μgL(-1)) in a flow-through field microcosm supplied with four contrasting natural waters. At the end of the exposure period, a 6-fold difference in the bioaccumulated Cu contamination levels was found among the four deployment sites, suggesting a significant control of the water quality on the metal bioaccumulation by aquatic mosses. In parallel, the so-called 'labile' Cu concentration for the same four field conditions was determined using a DGT device (Diffusive Gradient in Thin film). By coupling these DGT measurements and a cation competition model involving Ca(2+), Mg(2+), Na(+) and H(+), the time-dependent Cu concentrations in aquatic mosses were predicted; these simulation results were compared to the actual bioaccumulation of Cu in mosses. We found that any bioaccumulation model that ignores water characteristics is not suitable to predict the Cu accumulation by aquatic mosses under various water quality conditions. Instead, we found that our approach integrating DGT measurements and cationic composition was able to reproduce the Cu bioaccumulation kinetics by aquatic mosses for a wide range of water quality conditions. In conclusion, the DGT approach was demonstrated to be a dynamic in situ measuring technique that can be used as a surrogate of bioindicators if the cationic correction is taken into account., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The technique of diffusive gradient in thin film (DGT) is commonly used to assess metal contamination in natural waters. In this paper, we assess the effect of biofouling on DGT measured labile concentrations in water and investigate whether an additional nuclepore polycarbonate membrane on the surface of DGT devices can limit biofilm growth. Simultaneous field deployments of DGT equipped with and without the additional membrane in a canal receiving wastewater were compared. The effect of the biofilm was also assessed in controlled laboratory experiments, completed by the experimental determination of several metals diffusion coefficients in the hydrogel and membrane systems. The biofilms effect was problematic only from the 10th day of accumulation. Accumulation of some elements is highly biased by the presence of a thick biofilm (Zn, Ni, Cd). The polycarbonate membrane improved the quantification of Cd and Ni but adversely affects the quantification of Cr and Co. A kinetic model is proposed to explain the biofilm role on the DGT measurement. Depending on the metals of interest, it is possible to limit bias due to biofilms by using an additional polycarbonate membrane.

The effect of Ca, Zn, and dissolved natural organic matter (NOM) on waterborne Ni accumulation was investigated in a freshwater mussel. An enriched stable metal isotope tracer was required to measure the Ni uptake rate accurately. Zebra mussels were exposed to environmentally relevant concentrations of (62) Ni (from 0.5 to 8 µg/L) for 48 h in media spiked with Ca, Zn, or dissolved NOM. The (62)Ni uptake was inhibited by Ca (from 0.138 ± 0.021 to 0.061 ± 0.010 L/g/d for Ca concentrations ranging from 43 to 133 mg/L) and enhanced by Zn (from 0.051 ± 0.006 to 0.109 ± 0.007 L/g/d for Zn concentrations ranging from 6.6 to 38.3 µg/L). The mechanisms behind the synergistic effect of Zn remain unclear, yet it can be hypothesized that Ni uptake is facilitated by Zn-dependent transport sites. To formalize the effects of Ca and Zn, a model was proposed to express the Ni uptake rate as a function of the mussels' filtration rate and of Ca and Zn concentrations. The (62)Ni uptake increased at low NOM concentrations and decreased at higher concentrations. This could be explained by the influence of NOM on both the speciation of Ni and the filtration activity of mussels. At high NOM concentrations, a modification of the membrane's permeability might also have favored Ni uptake, although this was not clearly established in this study. Therefore, the effect of water composition on Ni bioavailability to zebra mussels cannot be predicted by competition and complexation models alone, because it also influences the animal's physiology., (Copyright © 2012 SETAC.)

This study is a first attempt to investigate the impact of urban contamination on metal tolerance of heterotrophic river biofilms using a short-term test based on β-glucosidase activity. Tolerance levels to Cu, Cd, Zn, Ni and Pb were evaluated for biofilms collected at three sites along an urban gradient in the Seine river (France). Metallic pollution increased along the river, but concentrations remained low compared to environmental quality standards. Biofilm metal tolerance increased downstream from the urban area. Multivariate analysis confirmed the correlation between tolerance and contamination and between multi-metallic and physico-chemical gradients. Therefore, tolerance levels have to be interpreted in relation to the whole chemical and physical characteristics and not solely metal exposure. We conclude that community tolerance is a sensitive biological response to urban pressure and that mixtures of contaminants at levels lower than quality standards might have a significant impact on periphytic communities., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The Water Framework Directive requires the development of biological tools which can act as early-warning indicators of a sudden increase (accidental pollution) or decrease (recovery due to prevention) of the chemical status of aquatic systems. River biofilms, which respond quickly to modifications of environmental parameters and also play a key part in the functioning of aquatic ecosystems, are therefore good candidates to monitor an increase or a decrease of water pollution. In the present study, we investigated the biological response of biofilms transplanted either upstream (recovery) or downstream (deterioration of exposure levels) the urban area of Paris (France). Both modifications of Cu community tolerance levels and of global bacterial and eukaryotic community structure using automated ribosomal intergenic spacer analysis (ARISA) fingerprints were examined 15 and 30 days after the transplantation. Cu tolerance levels of the heterotrophic component of biofilms were assessed using a short-term toxicity test based on β-glucosidase (heterotrophic) activity. Cu tolerance increased for biofilms transplanted upstream to downstream Paris (5-fold increase on day 30) and conversely decreased for biofilms transplanted downstream to upstream (8-fold decrease on day 30). ARISA fingerprints revealed that bacterial and eukaryotic community structures of transplanted biofilms were closer to the structures of biofilms from the transplantation sites (or sites with similar contamination levels) than to biofilms from their sites of origin. Statistical analysis of the data confirmed that the key factor explaining biofilm Cu tolerance levels is the sampling site and not the site of origin. It also showed that Cu tolerance levels are related to the global urban contamination (both metals and nutrients). The study shows that biofilms adapt fast to modifications of their surroundings. In particular, community tolerance varies quickly and reflects the new exposure levels only 15 days after transplantation. Those results support the use of biofilms as reliable early-warning indicators of diffuse urban contamination., (Copyright © 2011 Elsevier B.V. All rights reserved.)

This study investigates the ability of the biodynamic model to predict the trophic bioaccumulation of cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni) and zinc (Zn) in a freshwater bivalve. Zebra mussels were transplanted to three sites along the Seine River (France) and collected monthly for 11 months. Measurements of the metal body burdens in mussels were compared with the predictions from the biodynamic model. The exchangeable fraction of metal particles did not account for the bioavailability of particulate metals, since it did not capture the differences between sites. The assimilation efficiency (AE) parameter is necessary to take into account biotic factors influencing particulate metal bioavailability. The biodynamic model, applied with AEs from the literature, overestimated the measured concentrations in zebra mussels, the extent of overestimation being site-specific. Therefore, an original methodology was proposed for in situ AE measurements for each site and metal., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The biodynamic and saturation models offer promising lines of enquiry to predict the bioaccumulation of metals by aquatic organisms. However, in order to construct these models, the accumulation strategies have to be defined for each metal/organism couple in controlled conditions. This study aims at modelling the waterborne bioaccumulation of Ni and the influence of the water's geochemical properties on this process in a crustacean that is widely distributed in Europe, Gammarus pulex. In the laboratory, G. pulex was exposed to several Ni concentrations (from 0.001 to 100 mg L(-1)) in aquatic microcosms. Our results show that G. pulex is very tolerant to Ni (LC50(48 h)=477 mg L(-1) Ni). Time course experiments enabled the construction of a biodynamic model by determining the uptake (k(u)) and elimination (k(e)) rate constants. When the exposure concentration exceeded 1 mg L(-1) Ni, the metal uptake reached a maximum due to a limited number of binding sites for Ni. Therefore, the organism's maximal capacity to accumulate the metal (B(max)) and the half-saturation constant (K) were determined to establish the saturation model. We showed that the two models are comparable for the lowest exposure concentrations (<1 mg L(-1) Ni), with k(u)/k(e)=B(max)/K. Then, the bioaccumulation of Ni was recorded in waters exhibiting various concentrations of three major ions (Na(+), Mg(2+) and Ca(2+)). Only Ca had an inhibitory effect on the Ni uptake. This study reports for the first time the bioaccumulation of Ni in G. pulex. Because of its high tolerance to Ni and its high capacity to accumulate this metal, this crustacean could be used as an indicator of Ni bioavailability in freshwaters., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The biological response of periphyton chronically exposed to metals of urban origin (Cd, Ni and Zn) was investigated with a Pollution-Induced Community Tolerance (PICT) approach using a previously developed short-term toxicity test based on β-glucosidase (heterotrophic) activity. Periphyton was grown on plastic membranes immersed in indoor aquaria contaminated with metals at realistic contamination levels (0.3, 3 μg/l for Cd, 5, 50 μg/l for Ni, 20, 200 μg/l for Zn). After 3 weeks of exposure, biofilms' parameters (dry-weight, chlorophyll a concentration, heterotrophic activity) were analyzed and tolerance acquisition of the heterotrophic communities was assessed using the toxicity test. Modifications of bacterial and eukaryotic community structure were assessed with Automated Ribosomal Intergenic Spacer Analysis (ARISA). Effects of metal exposure were observed on biofilms parameters in the Cd and Zn experiments. Tolerance levels increased for both Cd-exposed biofilms, and for the high metal treatment biofilms in the Ni and Zn experiments. Analysis of the ARISA profiles showed that metal exposure affected the structure of both bacterial and eukaryotic communities. Moreover, Cd tolerance of the Zn-exposed heterotrophic communities was evaluated, which showed that the Zn-tolerant community (high metal treatment in the Zn experiment) also became tolerant to Cd (co-tolerance). The study shows that tolerance acquisition can be detected after exposure to environmental metal concentrations using β-glucosidase activity as an endpoint in short-term toxicity tests.

In order to obtain representative dissolved and solid samples from the aquatic environment, a spectrum of sampling methods are available, each one with different advantages and drawbacks. This article evaluates the use of discrete sampling and time-integrated sampling in illustrating medium-term spatial and temporal variation. Discrete concentration index (CI) calculated as the ratio between dissolved and solid metal concentrations in grab samples are compared with time-integrated concentration index (CI) calculated from suspended particulate matter (SPM) collected in sediment traps and labile metals measured by the diffusive gel in thin films (DGT) method, collected once a month during one year at the Seine River, upstream and downstream of the Greater Paris Region. Discrete CI at Bougival was found to be significantly higher than at Triel for Co, Cu, Mn, Ni and Zn, while discrete metal partitioning at Marnay was found to be similar to Bougival and Triel. However, when using time-integrated CI, not only was Bougival CI significantly higher than Triel CI, CI at Marnay was also found to be significantly higher than CI at Triel which was not observed for discrete CI values. Since values are time-averaged, dramatic fluctuations were smoothed out and significant medium-term trends were enhanced. As a result, time-integrated concentration index (CI) was able to better illustrate urbanization impact between sites when compared to discrete CI. The impact of significant seasonal phenomenon such as winter flood, low flow and redox cycles was also, to a certain extent, visible in time-integrated CI values at the upstream site. The use of time-integrated concentration index may be useful for medium- to long-term metal studies in the aquatic environment.

Biofilms are complex communities playing an important role in aquatic ecosystems. Automated ribosomal intergenic spacer analysis (ARISA) has been used successfully to explore biofilm bacterial diversity. However, a gap remains to be filled as regards its application to biofilm eukaryotic populations. The aim of this study is to use ARISA to detect eukaryotic population shifts in biofilm. We designed a new set of primers to focus specifically on the ITS1-5.8S-ITS2 region of diatoms and tested it on natural biofilms. Additionally, we tested universal primers, used previously to perform ARISA on fungal communities. Cloning and sequencing showed that the universal primer set amplified various eukaryotes, whereas the new set was diatom specific. The new set amplified a wider variety of diatoms. Therefore, the universal set is appropriate to study the general eukaryotic population shifts in biofilms, whereas the new set is more appropriate to study diatoms specifically. We used both primer sets, along with a bacterial set, to study the population shifts in natural river biofilms. Principal component analysis of the ARISA fingerprints revealed seasonal shifts that did not coincide for bacterial and eukaryotic communities. Therefore, the use of both eukaryotic and bacterial primers provides a useful insight to assess microbial succession in biofilms., (© 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

It is often difficult to evaluate the level of contamination in small urban rivers because pollution is mainly diffuse, with low levels of numerous substances. The use of a coupled approach using both chemical and biological measurements may provide an integrated evaluation of the impact of micro-pollution on the river. Zebra mussels were transplanted along a metal and organic pollution gradient in spring 2008. For two months, mussels and water samples were collected from two sites every two weeks and analyzed for metal and PAH content as well as water physicochemical parameters. Diffusive gradients in thin film (DGT) were also used to assess levels of labile metals. Exposure of mussels to contaminants and potential impact were evaluated using physiological indices and various biomarkers including condition index (CI), defense mechanisms (glutathione-S-transferase: GST), digestive enzymes (amylase and cellulase) and genotoxicity (micronucleus test: MN and comet assay: CA). For most contaminants, the water contamination was significantly higher downstream. Bioaccumulation in zebra mussels was related to water contamination in the framework of the biodynamic model, which allowed us to take into account the biological dilution that was caused by the growth of soft tissue downstream. Thus, metal influxes were on average two times higher downstream than upstream in particular for Zn, Cr, Cu and Cd. Significant differences in condition index were observed (final CI was 0.42 ± 0.03 downstream and 0.31 ± 0.03 upstream) reflecting a better food availability downstream. Moreover a significant decrease of GST activity and digestive enzymes activity in the cristalline style was observed downstream. Interpreting this decrease requires considering not only micro-pollution but also the trophic status related to the water's physicochemistry. The MN test and the CA on gill cells highlighted genotoxicity in mussels transplanted downstream compared to upstream., (© 2010 Wiley Periodicals, Inc. Environ Toxicol, 2010.)

A simple, low-cost and non-radioactive short-term toxicity test was developed to study the effects of urban metals on natural freshwater periphytic communities. β-glucosidase activity of natural freshwater biofilms collected in situ was chosen as an endpoint. Metals (Cd, Cu, Ni, Pb, and Zn) successfully inhibited bacterial enzymatic activity after a 1-h exposure enabling the calculation of EC(50). The EC(50) value of a biofilm sample varied with the Total Suspended Solids concentration (TSS) of the biofilm suspension, showing that EC(50) values (expressed as total added metal concentrations) are not representative of the bioavailable metal concentration during the toxicity test. For Cu, Cd, Ni, Zn and Pb, the EC(50) values increased linearly with the TSS concentration leading us to define a normalized EC(50): the value of the EC(50) divided by the corresponding TSS concentration. Normalized EC(50) proved to be a robust, reliable way to assess metal tolerance of a biofilm for Cd, Cu, Ni, Zn and Pb. Normalized EC(50) obtained, expressed as kg(metal)/g(TSS), varied between 0.2 to 7.6 for Cu, 1 to 8 for Cd, 1.8 to 92.3 for Ni, 1.8 to 76.6 for Zn and 25 to 189 for Pb.

The present study aims at investigating the effects of Zn, Ca, and dissolved organic carbon (DOC) on the waterborne Cd bioaccumulation of a freshwater bivalve (Dreissena polymorpha). Mussels were exposed for 48 h at 3 µg/L of Cd in different media. Their physiological activities were assessed by separately measuring the filtration rate in the same exposure water. Increased Zn (from 3 to 89 µg/L) and Ca (from 37 to 131 mg/L) concentrations in water led to a threefold and sevenfold reduction of Cd bioaccumulation, whereas the effect of DOC varied greatly depending on its concentration. At low DOC concentrations (from 0.2 to 1.1 mg/L), the uptake of Cd increased, whereas at higher concentrations (from 1.1 to 17.1 mg/L), the uptake decreased. The filtration activity was not strongly influenced by either Zn or Ca concentration, whereas it was modified in enriched DOC media in the same manner as Cd uptake. A competitive model was built to predict the waterborne uptake rate constant of Cd (k (u)) as a function of Zn and Ca concentrations in the water. Over the range of DOC concentrations we tested, organic matter was shown to influence Cd bioaccumulation in two ways: by modifying Cd speciation and thus its bioavailability and its interaction with the biological membrane, and by affecting the mussel's physiology and therefore its sensitivity to metal. The present study provides a useful means of adjusting the toxicokinetic constant to the water's physicochemical characteristics and proposes a unifying model that takes into account the different geochemical and biological influences on bioaccumulation., (Environ. Toxicol. Chem. 2010;29:2182-2189. © 2010 SETAC.)

Metal bioavailability depends on the presence of organic ligands in the water and on the concentrations of competitive cations. The present study aims at testing whether the diffusive gradient in thin films technique (DGT) could be used to take into account Cd speciation and its consequences on bioavailability in a bioaccumulation model and whether the influences of the Ca concentration and temperature also should be considered. Four kinetic experiments were conducted on Gammarus pulex: a calibration of Cd turnover rates and of the DGT lability in mineral water, a study of the influence f ethylenediaminetetraacetic acid (EDTA) and humic acids (HA) on uptake rates, and two experiments testing the influence of the Ca concentrations and temperature on Cd uptake clearance rates (ku). In mineral water, where Cd was considered fully labile, the ku was 0.46 L g⁻¹ d⁻¹, and the depuration rate was 0.032 d⁻¹. The initial Cd influxes were lowered significantly by additions of 10 μg L⁻¹ of EDTA or 10 mg L⁻¹ of HA in the water but not at 5 mg L⁻¹HA, even if DGT measurements proved that Cd formed Cd-HA complexes in that treatment. Increasing Ca concentrations lowered ku values, and a competitive inhibition model between Ca and Cd fitted the data. A 30% enhancement of k, values was observed when the temperature was increased by 8°C, which appeared comparatively as a weak effect. Thus, taking into account the metal speciation and the influence of the Ca concentration should improve Cd bioaccumulation modeling in amphipods. In freshwater, where metal bioavailability is reduced by the presence of dissolved organic matter, forecasting Cd waterborne uptake using the labile concentrations should allow robust comparisons between laboratory and field studies.

The present study investigated the effect of water cationic composition (Ca, Mg, Na, pH) on the bioaccumulation and elimination rates of copper by an aquatic moss (Fontinalis antipyretica), under laboratory conditions. For this purpose, mosses were exposed to copper at an environmentally relevant and usually non-toxic concentration (5 microg L(-1)) in natural waters where cationic composition and concentrations were varied. To describe copper bioaccumulation by aquatic mosses, a two-compartment model was the first-order kinetics, was developed and calibrated under a wide range of water cationic composition. Bioaccumulation rates of Cu in mosses were significantly reduced as the concentrations of competitive cations in solution increased. Hence, in hard-water, Ca and Mg cations play a protective role as they compete with Cu2+ ions for the absorption on transport sites at the organism-water interface. Based on the relationships between each major cation concentration and the exchange kinetics on mosses, the binding constants (K(Ci)(BL)) of each competing cations to the biological surfaces were derived. Using the present cationic-dependent kinetic model, it is now feasible to incorporate water cationic composition in the (re)interpretation of bryophytes contamination levels and in the (re)definition of Water Quality Criteria (WQC) as illustrated through two selected examples of biomonitoring programmes. In the framework of future national water quality guidelines revisions, a such flexible and mechanistic biomonitoring tool (integrating the protective effects of competing cations) may greatly improve the ability of regulators to derive site-specific Cu (metal) guidelines for protecting aquatic biota, while limiting the use of conservative assumptions.

Semipermeable membrane devices (SPMDs) previously spiked with performance reference compounds were exposed in wastewater. After 6 days of exposure, 13 polycyclic aromatic hydrocarbons (PAHs) were quantified in SPMDs. Exchange rate constants and time-weighted average (TWA) concentrations of SPMD-available PAHs in water were calculated. The bias of using SPMDs to estimate an actual TWA concentration if the concentration in water fluctuates, as can be expected in wastewater, was studied with numerical simulations. The bias increased with the exchange rate constant. However, most exchange rate constants evaluated in SPMDs exposed in wastewater were small enough for SPMDs to estimate a TWA concentration of PAHs even when the water concentration varied. TWA-SPMD-available concentrations were always below total dissolved (operationally defined as 0.7 microm) concentrations, indicating that part of the dissolved PAHs was not available for sampling. In situ partitioning coefficients K(DOC) were computed and found to be slightly higher than data from the literature. This confirms that only truly dissolved PAHs should be sampled by SPMDs in wastewater.

The present study investigates how dissolved organic matter (DOM) alters copper bioavailability at environmentally relevant concentrations (1-5 microg/L of dissolved copper, 1-4 mg/L of dissolved organic copper). A methodology combining two biological endpoints (short-term and steady-state bioaccumulation of copper by the aquatic moss Fontinalis antipyretica) and a sampling of labile copper with diffusion gradient in thin films (DGT) is proposed for batch experiments conducted with mineral water and various DOM, ethylenediaminetetra-acetic acid (EDTA), humic acid, and natural Seine River (France) extracts (hydrophobic and transphilic fractions). All types of DOM reduce the bioavailability of copper to aquatic mosses, and this reduction was more pronounced for the short-term biological endpoint, which was taken as being representative for environmental exposure. Labile copper sampled with DGT made it possible to estimate short-term bioaccumulation in the case of EDTA and natural Seine River extracts. With humic acid solutions, however, labile copper was lower than bioavailable copper. This result suggests that at realistic metal concentrations and with certain types of natural DOM, bioavailable copper might comprise not only inorganic copper but also some weak organic complexes. Hence, labile copper, in situ sampled with DGT, might not systematically overestimate bioavailable copper, as suggested previously on the basis of in vitro toxicity studies.

A combination of reverse osmosis (RO) concentration and DAX-8/XAD-4 resin adsorption techniques is used to isolate the various constituents of urban dissolved organic matter (DOM) from inorganic salts. Three fractions: hydrophobic (HPO), transphilic (TPI) and hydrophilic (HPI) accounting respectively for 35%, 20% and 45% of extracted carbon, are isolated from effluents of a major French wastewater treatment plant. This atypical DOC distribution, in comparison with natural water where the HPO fraction dominates, shows the significance of HPI fraction which often gets neglected because of extraction difficulties. A number of analytical techniques (elemental, spectroscopic: UV, FTIR) allow highlighting the weak aromaticity of wastewater effluent DOM (EfOM) due to fewer degradation and condensation processes and the strong presence of proteinaceous structures indicative of intense microbial activity. Copper toxicity in the presence of DOM is estimated using an acute toxicity test on Daphnia Magna (Strauss). Results reveal the similar protective role of each EfOM fraction compared to reference Suwannee river fulvic acid despite lower EfOM aromaticity (i.e. specific UV absorbance). The environmental implications of these results are discussed with respect to the development of site-specific water quality criteria.

Regulatory assessments of metal toxicity on freshwater organisms assume that toxic effects are caused by dissolved metals. In aquatic systems, organisms are exposed to both dissolved and particulate-bound metals. In this study, the chronic toxicity of dietary cadmium (Cd) on the reproduction and Cd body burden of Daphnia magna was investigated. Daphnids (<24 h) were successively exposed to dissolved Cd (8 h) and then to uncontaminated or contaminated algae (16 h) for 21 d. The results show a higher Cd burden in daphnids because of the addition of contaminated food and reveal that Cd uptake by D. magna from water and food was additive for the lowest Cd concentrations tested. Similar Cd distributions (cytosolic and insoluble fractions) were observed in the two groups of organisms, showing similar potential toxicity of Cd accumulated from the two exposure routes. Dietary Cd induces deleterious effects on D. magna reproduction. On the basis of Cd body burden of daphnids, the results support the claim that waterborne and dietary Cd exposures were additive in causing toxicity for Cd concentrations lower than 25 microg/L. At the highest Cd concentrations, the importance of dietary Cd on the daphnid contamination level decreases and confounding factors such as feeding rate reduction seem to appear, which induce an effect on neonate reproduction. In this study, we illustrate the need to take the dietary pathway into account in regulatory assessments and to establish effective concentrations with particulate-bound metals.

Sediments in aquatic ecosystems are often contaminated as a result of anthropogenic activities. Sediments and benthic organisms have been used to monitor trace metals contamination. However, due to the high variability of contaminant bioavailability, the attempt to link metal concentration in sediments and contamination of the organisms or ecotoxicological effect often lead to disappointing results. The technique of diffusive gradients in thin films (DGT) has been proposed as a relevant tool to study metal bioavailability, for example for accumulation in plants. In the present study, laboratory microcosm experiments were conducted with six contaminated sediments to compare metal accumulation in DGT and bioaccumulation in a chironomid (Chironomus riparius) for Cu, Cd and Pb . Metal accumulation in DGT was measured over time then modelled to determine two parameters of the dynamic response of the metals to DGT deployment: the size of the particulate labile pool and the kinetic of the solid-dissolved phase exchange. The mobility of metals was found metal and sediment dependent. A significant relationship between metal accumulated in DGT and bioaccumulated in chironomids was found for Cu and Pb. However, total metals in sediments were the best predictors of bioaccumulation. Nevertheless, the knowledge of the metals dynamic enhanced our ability to explain the different biological uptake observed in sediments of similar total metal concentrations.

Diffusive Gradient in Thin Films (DGT) and Semi-Permeable Membrane Devices (SPMDs) were deployed in the Seine river basin in order to assess labile metals and truly dissolved Polycyclic Aromatic Hydrocarbons. We show that the tools are reliable in aquatic environments to assess the speciation of dissolved contaminants and hence provide a good insight into the potential bioavailability of contaminants. The deployment of the DGT and SPMDs in contrasting environments in the Seine river basin allowed distinction to be made of availability of contaminants between headwater streams and much more impacted river reaches and an assessment of bioavailability. At the stations under urban influence, the impact of dissolved organic matter on both copper and PAHs bioavailability is less pronounced than at upstream stations, where humic substances dominate.

Periphytic biomass has an important influence on the water quality of many shallow streams. The purpose of this paper is to synthesize the knowledge obtained on periphyton during the PIREN Seine research program. Periphyton was sampled using chl a measurements by acetone extraction and oxygen measurements with microelectrodes. The experiments reveal the presence of an important fixed biomass ranging between 123 and 850 mgchl a m(-2) and the mean gross production (photosynthesis) is shown to range between 180 and 315 mgC m(-2) h(-1). An independent approach was performed using the ProSe model, which simulates transport and biogeochemical processes in 22 km of the Grand Morin stream. A strong agreement between in situ measurements and the model results was obtained. The gross production obtained using ProSe is 220 mgC m(-2) h(-1) for the periphyton, which matches the experimental data. Although the net photosynthetic activity of the phytoplankton (0.84 gC gC(-1) d(-1)) is higher than the periphytic one (0.33 gC gC(-1) d(-1)), the absolute periphytic activity is greater since the mean biomass (3.4 gC m(-)(2)) is 10 times higher than the phytoplanktonic one (0.3 gC m(-2)), due to the short residence time of the water body (1.5d).

Seven metals (Cd, Co, Cr, Cu, Fe, Ni and Pb) were monitored at the Seine-Aval wastewater treatment plant during 6 sampling campaigns in April 2004. Particulate and dissolved metals have been measured in 24h composite samples at each treatment stage (primary settling, secondary activated sludge and tertiary flocculation by FeCl(3)). In addition, the diffusive gradient in thin film technique (DGT) was used to determine the dissolved inert and labile metal fraction. Although all treatment stages were able to decrease particulate metals concentrations in wastewater, most dissolved metals concentrations were mainly affected during primary settling. This unexpected result was attributed to tertiary sludge filtrate recirculation. Metals added via the FeCl(3) reagent at the tertiary treatment were shown to lower the overall Cr removal from wastewater and to enrich Ni in effluents. The plant operating conditions (recirculation and reagent addition) appear therefore as important as treatment processes for the metals removal. Total metal fluxes were highly decreased by the whole treatment plant for Cd, Cr, Cu and Pb and to a lesser extend for Co and Ni. However, the labile metal fluxes were poorly decreased for Cu (18%), not significantly decreased for Ni and increased for Fe. The labile fraction of Cd, Co and Cr was not detectable at any stage of the plant. Discharged labile fluxes, at least for Ni, were potentially significant compared to the labile metal fluxes in the river measured downstream the plant. Treated urban wastewater discharges should be carefully considered as a possible source of bioavailable trace metals.

The concentrations of dissolved trace elements (Li, B, Mn, Cu, As, Rb, Sr, Mo, Cd, Ba, Pb) in the Marne and Seine rivers in the Paris urban area were monitored over a 2-year period. The resulting data indicated moderate contamination of waters by the most toxic elements (Cu, As, Cd and Pb). The River Marne upstream and the River Seine downstream of the city of Paris displayed similar concentrations. However higher fluxes of trace elements were observed in the Seine than in the Marne due to their different discharges. Li, B, Rb, Sr and Ba concentrations were correlated with river discharge and concentrations were higher during high river flow. This was interpreted as a dilution by discharge from a major natural or anthropogenic source. Mn, Cu, Mo, Cd and Pb concentrations were not correlated with discharge. Dissolved Mn, Cu and Cd increased rapidly in summer, whereas the concentration of Mo decreased. These variations were attributed to redox processes. During summer when the dissolved oxygen concentrations decrease, Mn, Cu, Cd and Pb are released into solution whereas Mo is immobilised. Like metals, variations in arsenic contents were not linked with discharge. Its similarity with phosphate distribution suggests similar controls involving phytoplankton uptake and release from sediments through organic matter mineralization.

This paper aims to evaluate the performance of the diffusive gradient in thin film technique (DGT) as a speciation tool for metals in wastewater. The validity of metal sampling by DGT in wastewater was checked. DGT was used in parallel with the Daphnia magna acute toxicity test in order to obtain information on the speciation of copper and cadmium in diluted and spiked filtered wastewater (raw and treated) from two treatment plants. Combining the chemical (DGT) and the biological methods (D. magna toxicity test) allowed metal to be fractionated into inorganic, labile organic and inert organic metal. Copper was mainly found as inert organic complexes, whereas the major part of cadmium was found to be labile organic complexes. The proportion of inert organic copper complexes was higher in the presence of treated wastewater than in raw wastewater. The use of restricted gels in DGT devices discriminated more labile organic cadmium than labile organic copper, indicating that cadmium weak ligands have more complex structures than copper weak ligands. In our experimental conditions (i.e. a high metal to ligand ratio), DGT, even equipped with restricted gels, was able to accumulate labile organic complexes. This result highlights that the ecotoxicological interpretation of DGT measurement should be considered carefully. DGT is a reliable tool to assess the chemical characteristics of metals (i.e. reactivity) in wastewater, but it does not ensure that only inorganic metal is measured.

This paper presents a methodology for assessing the variability of biodegradable chemical oxygen demand (COD) fractions in urban wastewaters. Thirteen raw wastewater samples from combined and separate sewers feeding the same plant were characterised, and two optimisation procedures were applied in order to evaluate the variability in biodegradable fractions and related kinetic parameters. Through an overall optimisation on all the samples, a unique kinetic parameter set was obtained with a three-substrate model including an adsorption stage. This method required powerful numerical treatment, but improved the identifiability problem compared to the usual sample-to-sample optimisation. The results showed that the fractionation of samples collected in the combined sewer was much more variable (standard deviation of 70% of the mean values) than the fractionation of the separate sewer samples, and the slowly biodegradable COD fraction was the most significant fraction (45% of the total COD on average). Because these samples were collected under various rain conditions, the standard deviations obtained here on the combined sewer biodegradable fractions could be used as a first estimation of the variability of this type of sewer system.

In order to better asses the influence of organic matter on the bioavailability of hydrophobic organic contaminants, the effect of algae and POM of bacterial origin on the bioaccumulation of benzo[a]pyrene in Daphnia magna was evaluated. The bioaccumulation was monitored with increasing concentrations of particulate organic matter (POM) and dissolved organic matter (DOM). In all experiments, the presence of POM greatly reduced the bioaccumulation of benzo[a]pyrene. The reduction was more pronounced in the presence of algae, for which we observed a 99%-reduction effect in the presence of 6 x10 (5) cell/mL (equivalent to 5.3 mg C/L). The bioaccumulation of benzo[a]pyrene was decreased by 49% by organic matter of bacterial origin at 4.7 mg C/L. Assuming that benzo[a]pyrene was partitioned between water, DOM and POM and supposing that D. magna accumulated free benzo[a]pyrene via respiration and POM-bond benzo[a]pyrene via ingestion, bioaccumulation data allowed to estimate the dietary uptake rate of benzo[a]pyrene as well as partitioning coefficients K(POC) and K(DOC). Despite the ingestion of contaminated particles, we could not observe any dietary uptake of benzo[a]pyrene in daphnids. We verified, as usually supposed, that the bioaccumulation of benzo[a]pyrene to D. magna occurs mainly via direct contact. Very high partitioning coefficients (log K(POC) between 5.2 and 6.2) were estimated. This study pointed out the great influence of biogenic organic matter on the fate and the bioavailability of benzo[a]pyrene in aquatic ecosystems.

The biodegradation of two substrates and the ability of dissolved organic matter (DOM) to influence benzo[a]pyrene bioavailability as DOM biodegradation progressed were monitored in reactors. Substrates were composed of algae extracts and an artificial substrate that mimics raw wastewater, which were considered to be autochthonous and anthropogenic allochthonous models for DOM, respectively. The soluble microbial products formed during biomass activity were also studied. The aromaticity of DOM was investigated with specific ultraviolet absorbance. Partitioning coefficients between DOM and benzo[a]pyrene, K(DOC)(biol), were biologically determined by means of 4-h bioaccumulation experiments on Daphnia magna. Parent and degraded substrates always significantly reduced the bioaccumulation of benzo[a]pyrene at environmental DOM concentrations. Soluble microbial products also significantly affected the benzo[a]pyrene bioaccumulation. K(DOC)(biol) ranged between 2 x 10(4) and 4 x 10(5) L/kg. As the artificial wastewater biodegraded, DOM aromaticity increased, as did K(DOC)(biol). During the biodegradation of algae extract DOM, K(DOC)(biol) increased, whereas their aromaticity slightly decreased.

This study investigates the relevance of the diffusion gradient in thin films technique (DGT) to measure copper's induced lethality on Daphnia magna in natural water spiked with various organic ligands. Ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), and glycine were used as artificial organic ligands in controlled solutions of mineral water. With EDTA, DGT measurement makes it possible to predict the toxicity of the mixture because Cu-EDTA complexes are inert, whereas DGT is of no help in the case of NTA, because Cu-NTA complexes are fully labile. The Cu-glycine complexes appear as partly labile and toxic. Humic acids as well as fresh and aged algae extracts also were used as models for natural dissolved organic matters. All three of them form copper complexes that are not toxic to Daphnia magna and appear as partly labile with open-pored DGT. However, the use of restricted gels in DGT greatly reduces the contribution of labile complexes, at least for humic acids and aged algae copper complexes. The DGT with restrictive gels, therefore, appears to be a powerful tool for measuring bioavailable copper in natural water bodies, especially when the dissolved organic matter mostly is of humic origin. The DGTs potential ability to mimic the lability induced by the biological uptake also is discussed.

As a preliminary study of the influence of urban organic matter on the bioavailability of polycyclic aromatic hydrocarbons (PAHs), the effect of different types of dissolved organic matter (DOM) on the bioaccumulation of fluoranthene, pyrene, or benzo[a]pyrene in Daphnia magna was studied. Commercial humic substances, DOM from the aeration basin of a wastewater treatment plant, and highly biodegradable DOM (algae or animal extracts) were tested. The bioaccumulation of benzo[a]pyrene was reduced by each DOM (up to 80% reduction with humic substances). Pyrene bioaccumulation was also decreased by each DOM to a lesser extent. Fluoranthene bioaccumulation was affected by the presence of humic acids only. In each experiment, the solution containing humic DOM led to the lowest bioaccumulation. Supposing that only dissolved PAHs were bioavailable, the reduction of bioaccumulation allowed a biological estimate of the partition coefficients of DOM and PAH, K(DOC). The estimated coefficients were positively related to the aromaticity of DOM and negatively related to its biodegradability.

The biodegradability of water samples is usually estimated with bioassays under oxic conditions. In order to overcome some of the drawbacks linked to the incubation of the samples in aerobic batches, a new protocol is proposed and tested, which is based on an organic carbon (OC) balance after a 45 days incubation under anoxic conditions with excess nitrate. The biodegradable fractions of organic matter obtained with the anoxic protocol are slightly lower than those obtained under oxic conditions. Several possible reasons for a systematic underestimation of the biodegradable organic matter under anoxic conditions are evaluated and discussed: a reduced microbial metabolic potential, significantly reduced degradation rates for the slowly biodegradable organic matter, an additional production of refractory organic compounds during the incubation, or the inhibition of the recycling of the organic matter stored in bacterial biomass. Nevertheless, the 7% difference observed on the biodegradable total OC estimations keeps low enough so that the anoxic protocol can be proposed as a convenient alternative to the oxic one.

The speciation of metal species extracted by diffusion gradient in thin films (DGT) devices during experimental deployments in simple metal-ligand synthetic solutions was numerically modeled, analyzed at steady state, and simulated in the dynamic regime. The modeled speciation of two well-known complexes (Cu-citrate and Cu-EDTA) are in good agreement with experimental data obtained in NaNO3 solutions. For any metal complex, the rate at which the metal accumulates on the chelating resin is proportional to the concentration of free metal in solution plus a fraction of the metal complex concentration in solution equivalent to xiD(ML)/D(M). D(M) and D(ML) are the diffusion coefficients of the free and metal complex, respectively, and xi characterizes the complex lability: it is defined as the fraction of metal complex lost when diffusing from the bulk solution to the chelating resin. Numerical simulations were used to explore the variations of xi as a function of several operational and chemical parameters. Lability increases when the dissociation rate constant or the residence time of the metal complex within the hydrogel increase (i.e., by either increasing the thickness of the hydrogel or decreasing D(ML)). Overall, the fraction of metal complex extracted by the DGT decreases when D(ML) decreases, which confirms the ability of diffusion-restrictive hydrogels to separate the free metal fraction from complex solutions. Although the DGT model does not exactly comply with the voltammetric formalism, the lability criteria deltatau(1/2) used with voltammetric macroelectrodes is similar to the lability criteria xi, except that it is valid for high metal-ligand ratio. To determine free metal concentrations in aquatic systems using DGT, it is necessary to account for the presence of numerous ligands with different complexing properties. Soon, the numerical model will be improved to include additional ligands competing for the metal, and new experiments will be designed to discriminate between different labile complexes. Experimental results obtained with different DGT devices (of different hydrogel thicknesses, for example) could be interpreted in terms of conditional kinetic characteristics of the dominant metal-ligand complex in solution.

Two different re-oxygenation techniques (aeration and hydrogen peroxide addition) were compared in respirometric experiments. As similar results were obtained in both cases, it was concluded that the addition of hydrogen peroxide does not modify the oxygen uptake rate of the biomass, under either endogenous or feeding conditions. It was hypothesized that under those experimental conditions (inhibition of nitrification with ATU), hydrogen peroxide alters neither the biomass metabolism nor the biodegradability of the tested substrates. The oxygen uptake rates obtained with the aeration system were often more scattered due to the adhesion of fine bubbles after the switch off of the aeration. Moreover, the transfer rate of oxygen to the solution is faster in the case of hydrogen peroxide addition.

Eutrophication is the enrichment of water bodies with plant nutrients and precursors, typically nitrogen, phosphorus, and organic matter. There exists a "natural" and slow eutrophication, which, over geological times, turns a lake into a marsh and then dries it entirely. Today, however, eutrophication is mostly referred to the human process that "results in the stimulation of an array of symptomatic changes, among which increased production of algae and macrophytes, deterioration of water quality and other symptomatic changes are found to be undesirable and interfere with water uses" as defined by the OECD in 1982. This undesirable process is observed mostly in enclosed water bodies, such as lakes, but also in some rivers, some estuaries, and some coastal zones. In most freshwater systems, phosphorus has been identified as the "limiting nutrient" to phytoplankton development. This nutrient is brought to aquatic environments from rock weathering, soil leaching, and rain (natural sources), but also and mostly from agricultural runoff and domestic sewage. Some food processing industries (meat, vegetables, cheese processing) also contribute significantly to the phosphorus budget, even though the pollution may be due to floor and utensil cleaning rather than to direct food wastes., (Copyright 2001 International Life Sciences Institute.)