Avoidance behaviour of aquatic macroinvertebrates for real-time detection of micropollutant surge in wastewater effluents.

Micropollutants are regularly detected at the outlets of wastewater treatment plants (WWTPs). Across urban and industrial WWTPs, monitoring directives only require assessment for a handful of chemicals via sampling methods that fail to capture the temporal variability in micropollutant discharge. In this study, we develop a biotest for real-time on-line monitoring of micropollutant discharge dynamics in WWTPs effluents. The selected biomonitoring device ToxMate uses videotracking of invertebrate movement, which was used to deduce avoidance behaviour of the amphipod Gammarus fossarum. Organism conditioning was set up to induce a state of minimal locomotor activity in basal conditions to maximise avoidance signal sensitivity to micropollutant spikes. We showed that with a standardised protocol, it was possible to minimise both overall movement and sensitivity to physio-chemical variations typical to WWTP effluents, as well as capture the spikes of two micropollutants upon exposure (copper and methomyl). Spikes in avoidance behaviour were consistently seen for the two chemicals, as well as a strong correlation between avoidance intensity and spiked concentration. A two-year effluent monitoring case study also illustrates how this biomonitoring method is suitable for real-time on-site monitoring, and shows a promising non-targeted approach for characterising complex micropollutant discharge variability at WWTP effluents, which today remains poorly understood.

Effects of diclofenac on sentinel species and aquatic communities in semi-natural conditions.

Due to the potential hazard of diclofenac on aquatic organisms and the lack of higher-tier ecotoxicological studies, a long-term freshwater mesocosm experiment was set up to study the effects of this substance on primary producers and consumers at environmentally realistic nominal concentrations 0.1, 1 and 10 µg/L (average effective concentrations 0.041, 0.44 and 3.82 µg/L). During the six-month exposure period, the biovolume of two macrophyte species (Nasturtium officinale and Callitriche platycarpa) significantly decreased at the highest treatment level. Subsequently, a decrease in dissolved oxygen levels was observed. High mortality rates, effects on immunity, and high genotoxicity were found for encaged zebra mussels (Dreissena polymorpha) in all treatments. In the highest treatment level, one month after the beginning of the exposure, mortality of adult fish (Gasterosteus aculeatus) caused effects on the final population structure. Total abundance of fish and the percentage of juveniles decreased whereas the percentage of adults increased. This led to an overall shift in the length frequency distribution of the F1 generation compared to the control. Consequently, indirect effects on the community structure of zooplankton and macroinvertebrates were observed in the highest treatment level. The No Observed Effect Concentration (NOEC) value at the individual level was < 0.1 µg/L and 1 µg/L at the population and community levels. Our study showed that in more natural conditions, diclofenac could cause more severe effects compared to those observed in laboratory conditions. The use of our results for regulatory matters is also discussed.

Nongenetic inheritance of increased Cd tolerance in a field Gammarus fossarum population: Parental exposure steers offspring sensitivity.

While the phenomenon of increased population tolerance to environmental contaminants has been widely reported for aquatic animal species living in contaminated ecosystems, the question of the nature of the tolerance, i.e., genetic adaptation or phenotypic plasticity, has not yet been studied to the same degree and remains under debate when explaining the variability of toxicological sensitivity among field populations. The occurrence of both processes challenges the ecological risk assessment (ERA) of chemicals, questioning the relevance of toxicity assessment procedures based on bioassays employing organisms from naive reference populations. Nonetheless, given that genetic adaptation and phenotypic acclimation are based on different mechanisms, with different persistence periods over time when exposure is stopped, gaining insight into the mechanism of tolerance to contaminants of field populations is a major concern for ERA. In this context, this study aimed to test whether genetic adaptation underlies the tolerance to cadmium (Cd) in a field Gammarus fossarum population identified in a previous study. Breeding and exposure experiments conducted in laboratory conditions revealed that: (1) first and second cohorts of neonates produced from field-collected parents present an increased Cd tolerance, which is explained by a reduction of between-brood variability of Cd sensitivity with an absence of sensitive broods compared to two reference populations rather than by the existence of more tolerant individuals; and (2) Cd tolerance was no longer evident for broods of the third cohort produced by these breeders when maintained under Cd-free conditions in the laboratory. Hence, this study drew the striking conclusion that the observation of Cd tolerance inheritance within the field population studied does not demonstrate genetic adaptation, but it results from an effect of parental exposure inducing transgenerational plasticity, which deeply influences the variability of population sensitivity. To our knowledge, this study is only the second report of increased tolerance to contaminants in a field population explicitly attributed to the phenomenon of transgenerational acclimation.

Identification, expression, and endocrine-disruption of three ecdysone-responsive genes in the sentinel species Gammarus fossarum.

Taking advantage of a large transcriptomic dataset recently obtained in the sentinel crustacean amphipod Gammarus fossarum, we developed an approach based on sequence similarity and phylogenetic reconstruction to identify key players involved in the endocrine regulation of G. fossarum. Our work identified three genes of interest: the nuclear receptors RXR and E75, and the regulator broad-complex (BR). Their involvement in the regulation of molting and reproduction, along with their sensitivity to chemical contamination were experimentally assessed by studying gene expression during the female reproductive cycle, and after laboratory exposure to model endocrine disrupting compounds (EDCs): pyriproxyfen, tebufenozide and piperonyl butoxide. RXR expression suggested a role of this gene in ecdysis and post-molting processes. E75 presented two expression peaks that suggested a role in vitellogenesis, and molting. BR expression showed no variation during molting/reproductive cycle. After exposure to the three EDCs, a strong inhibition of the inter-molt E75 peak was observed with tebufenozide, and an induction of RXR after exposure to pyriproxyfen and piperonyl butoxide. These results confirm the implication of RXR and E75 in hormonal regulation of female reproductive cycles in G. fossarum and their sensitivity towards EDCs opens the possibility of using them as specific endocrine disruption biomarkers.

Comparison in waterborne Cu, Ni and Pb bioaccumulation kinetics between different gammarid species and populations: Natural variability and influence of metal exposure history.

Kinetic parameters (uptake from solution and elimination rate constants) of Cu, Ni and Pb bioaccumulation were determined from two Gammarus pulex and three Gammarus fossrum wild populations collected from reference sites throughout France in order to assess the inter-species and the natural inter-population variability of metal bioaccumulation kinetics in that sentinel organism. For that, each population was independently exposed for seven days to either 2.5µgL-1 Cu (39.3nM), 40µgL-1 Ni (681nM) or 10µgL-1 Pb (48.3nM) in laboratory controlled conditions, and then placed in unexposed microcosms for a 7-day depuration period. In the same way, the possible influence of metal exposure history on subsequent metal bioaccumulation kinetics was addressed by collecting wild gammarids from three populations inhabiting stations contaminated either by Cd, Pb or both Pb and Ni (named pre-exposed thereafter). In these pre-exposed organisms, assessment of any changes in metal bioaccumulation kinetics was achieved by comparison with the natural variability of kinetic parameters defined from reference populations. Results showed that in all studied populations (reference and pre-exposed) no significant Cu bioaccumulation was observed at the exposure concentration of 2.5µgL-1. Concerning the reference populations, no significant differences in Ni and Pb bioaccumulation kinetics between the two species (G. pulex and G. fossarum) was observed allowing us to consider all the five reference populations to determine the inter-population natural variability, which was found to be relatively low (kinetic parameters determined for each population remained within a factor of 2 of the minimum and maximum values). Organisms from the population exhibiting a Pb exposure history presented reduced Ni uptake and elimination rate constants, whereas no influence on Ni kinetic parameters was observed in organisms from the population exhibiting an exposure history to both Ni and Pb. Furthermore Pb bioaccumulation kinetics were unaffected whatever the condition of pre-exposure in natural environment. Finally, these results highlight the complexity of confounding factors, such as metal exposure history, that influence metal bioaccumulation processes and showed that pre-exposure to one metal can cause changes in the bioaccumulation kinetics of other metals. These results also address the question of the underlying mechanisms developed by organisms to cope with metal contamination.

Assessing the relevance of a multiplexed methodology for proteomic biomarker measurement in the invertebrate species Gammarus fossarum: A physiological and ecotoxicological study.

Recently, a protein sequence database was built specifically for the sentinel non-model species Gammarus fossarum using a proteogenomics approach. A quantitative multiplexed targeted proteomics assay (using Selected Reaction Monitoring mass spectrometry) was then developed for a fast and simultaneous quantification of dozens of biomarker peptides specific of this freshwater sentinel crustacean species. In order to assess the relevance of this breakthrough methodology in ecotoxicology, the response patterns of a panel of 26 peptides reporting for 20 proteins from the Gammarus fossarum proteome with putative key functional roles (homeostasis, osmoregulation, nutrition, reproduction, molting,…) were recorded through male and female reproductive cycles and after exposure to environmental concentrations of cadmium and lead in laboratory-controlled conditions. Based on these results, we validated the implication of annotated vtg-like peptides in the oogenesis process, and the implication of Na+/K+ ATPase proteins in the molt cycle of organisms. Upon metal (cadmium and lead) contamination, peptides belonging to proteins annotated as involved in antioxidant and detoxification functions, immunity and molting were significantly down-regulated. Overall, this multiplex assay allowed gaining relevant insights upon disruption of different main functions in the sentinel species Gammarus fossarum. This breakthrough methodology in ecotoxicology offers a valid and high throughput alternative to currently used protocols, paving the way for future practical applications of proteogenomics-derived protein biomarkers in chemical risk assessment and environmental monitoring.

Active biomonitoring for assessing effects of metal polluted sediment resuspension on gammarid amphipods during fluvial traffic.

The resuspension of polluted sediments by boat traffic could release substantial amounts of metals to the water column, affecting at the same time their bioavailability. In order to characterize the impact of sediment resuspensions on biota, caged amphipods have been deployed on three different channelized watercourses in Northern France. Firstly, the biological responses of transplanted freshwater gammarid amphipods, Gammarus fossarum, described by trace metal accumulation, feeding and reproduction activities were quite similar for the three water courses despite the differences of metal contamination and navigability. Secondly, the concentrations of metals accumulated in gammarids never exceeded the contamination thresholds previously defined for Co, Cu, Cr and Zn. Values were in the same order of magnitude whatever the studied site despite: (i) large differences noticed in the sediment quality and (ii) some concentrations in the overlying waters exceeding the Environmental Quality Standards (EQS) defined by the Water Framework Directive. Conversely, Pb was highly bioaccumulated with values systematically exceeding the threshold value whatever the site. Therefore, the impact of navigation cannot be proved and the difference between the 3 monitoring periods is rather attributed to environmental variability, probably linked to the seasonality. Moreover, this study also confirms that organisms sampled from a local population in the vicinity of the three studied watercourses could be used as test organisms, leading to similar results than the ones obtained with reference gammarids initially used for developing all the biological responses. This would simplify and then promote the development of studies based on gammarid amphipod, G. fossarum, as bioindicators.

Environmental relevance of laboratory-derived kinetic models to predict trace metal bioaccumulation in gammarids: Field experimentation at a large spatial scale (France).

Kinetic models have become established tools for describing trace metal bioaccumulation in aquatic organisms and offer a promising approach for linking water contamination to trace metal bioaccumulation in biota. Nevertheless, models are based on laboratory-derived kinetic parameters, and the question of their relevance to predict trace metal bioaccumulation in the field is poorly addressed. In the present study, we propose to assess the capacity of kinetic models to predict trace metal bioaccumulation in gammarids in the field at a wide spatial scale. The field validation consisted of measuring dissolved Cd, Cu, Ni and Pb concentrations in the water column at 141 sites in France, running the models with laboratory-derived kinetic parameters, and comparing model predictions and measurements of trace metal concentrations in gammarids caged for 7 days to the same sites. We observed that gammarids poorly accumulated Cu showing the limited relevance of that species to monitor Cu contamination. Therefore, Cu was not considered for model predictions. In contrast, gammarids significantly accumulated Pb, Cd, and Ni over a wide range of exposure concentrations. These results highlight the relevance of using gammarids for active biomonitoring to detect spatial trends of bioavailable Pb, Cd, and Ni contamination in freshwaters. The best agreements between model predictions and field measurements were observed for Cd with 71% of good estimations (i.e. field measurements were predicted within a factor of two), which highlighted the potential for kinetic models to link Cd contamination to bioaccumulation in the field. The poorest agreements were observed for Ni and Pb (39% and 48% of good estimations, respectively). However, models developed for Ni, Pb, and to a lesser extent for Cd, globally underestimated bioaccumulation in caged gammarids. These results showed that the link between trace metal concentration in water and in biota remains complex, and underlined the limits of these models, in their present form, to assess trace metal bioavailability in the field. We suggest that to improve model predictions, kinetic models need to be complemented, particularly by further assessing the influence of abiotic factors on trace metal uptake, and the relative contribution of the trophic route in the contamination of gammarids.

Zebra mussel as a new tool to show evidence of freshwater contamination by waterborne Toxoplasma gondii.

AIMS: The objective of this study was to evaluate if freshwater bivalves can be used to detect the presence of Toxoplasma gondii in water bodies. METHODS AND RESULTS: Zebra mussels (Dreissena polymorpha) were caged for 1 month upstream and downstream of the discharge points of wastewater treatment plants (WWTPs). Physiological status was assessed to assure good health of bivalves during transplantation. The presence of T. gondii was investigated in mussel tissues by qPCR. In autumn, T. gondii was detected in mussels caged downstream of the discharge points of two WWTPs. In spring, it was detected upstream of one WWTP. CONCLUSIONS: For the first time, T. gondii DNA has been shown in a continental mollusc in environmental conditions. This highlights the interest of an active approach that could be applied independently of the presence or accessibility of autochthonous populations, and underlines the presence of T. gondii in natural waters under pressure of WWTP discharge at a certain time of the year. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that transplanted zebra mussels could be used as biosamplers to reveal contamination of freshwater systems by T. gondii.

Evolution of cadmium tolerance and associated costs in a Gammarus fossarum population inhabiting a low-level contaminated stream.

Deciphering evolutionary processes occurring within long-term contaminated wild populations is essential for the ecological risk assessment of persistent chemical contaminations. Using field populations of Gammarus, a commonly-used genus in aquatic ecotoxicology, the present study sought to gain insights into the extent to which long-term exposure to metals in the field could effectively lead to shifts in toxicological sensitivities. For this, we identified a Gammarus population inhabiting a stream contaminated by cadmium (Cd). We compared the Cd-exposure and Cd-sensitivity of this population to those of five reference populations. Active biomonitoring determined in different years and seasons that significant levels of Cd were bioavailable in the contaminated site. Laboratory sensitivity tests under common garden conditions established that this long-term field exposure led to the development of a moderate Cd tolerance, which was maintained after a 3-week acclimatization in the laboratory, and transmitted to offspring produced under clean conditions. The potential physiological costs of tolerance were assessed by means of feeding rate measurements (in the laboratory and in situ). They revealed that, unlike for reference populations, the feeding activity of organisms from the tolerant population was greatly decreased when they were maintained under laboratory conditions, potentially indicating a high population vulnerability to environmental perturbations. Because dissolved Cd concentrations in water from the contaminated site were low (averaging 0.045 µg L(-1)) and below the current European environmental quality standard for Cd for inland surface waters (fixed at 0.08 µg L(-1) in soft water environments), this case study sheds light onto the extent to which current environmental quality standards are protective against potential adverse outcomes of adaptive and micro-evolutionary processes occurring in contaminated environments.

A biodynamic model predicting waterborne lead bioaccumulation in Gammarus pulex: Influence of water chemistry and in situ validation.

Metals bioaccumulated in aquatic organisms are considered to be a good indicator of bioavailable metal contamination levels in freshwaters. However, bioaccumulation depends on the metal, the species, and the water chemistry that influences metal bioavailability. In the laboratory, a kinetic model was used to describe waterborne Pb bioaccumulated in Gammarus pulex. Uptake and elimination rate constants were successfully determined and the effect of Ca(2+) on Pb uptake was integrated into the model. Thereafter, accumulated Pb concentrations in organisms were predicted with the model and compared with those measured in native populations from the Seine watershed (France). The predictions had a good agreement with the bioaccumulation levels observed in native gammarids and particularly when the effect of calcium was considered. To conclude, kinetic parameters experimentally derived for Pb in G. pulex are applicable in environmental conditions. Moreover, the consideration of the water's chemistry is crucial for a reliable interpretation of bioaccumulation.

Temporal patterns of digestive enzyme activities and feeding rate in gammarids (Gammarus fossarum) exposed to inland polluted waters.

The aim of this study was to use digestive enzyme activities in Gammarus fossarum as biomarkers during active biomonitoring. Standardised gammarids were transplanted for 7 days to five sites in the Riou Mort watershed contaminated by polymetallic pollution. This experiment was conducted on seven different dates from February 2009 to June 2010. Feeding rates were tracked, along with amylase, cellulase and trypsin activities. We found that feeding rate and digestive capacity were reduced in the most polluted site, "Joany," in comparison with the reference site "Up.Lot". The results suggested that trypsin was more sensitive than the other two carbohydrases. In the four other sites, seasonal differences were observed during the 2yr but no clear pattern can be established. This study highlights the ability of G. fossarum to demonstrate environmental disturbances and suggests the use of a caging process in certain seasons. Caging organisms and feeding ad libitum is advantageous, as it reduces inter-individual variability and removes dependence on the native food fluctuations. However, confounding factors other than temperature were present, and the interpretation of digestive enzyme activities is complex.

Effect of water quality and confounding factors on digestive enzyme activities in Gammarus fossarum.

The feeding activity and subsequent assimilation of the products resulting from food digestion allow organisms to obtain energy for growth, maintenance and reproduction. Among these biological parameters, we studied digestive enzymes (amylase, cellulase and trypsin) in Gammarus fossarum to assess the impact of contaminants on their access to energy resources. However, to enable objective assessment of a toxic effect of decreased water quality on an organisms' digestive capacity, it is necessary to establish reference values based on its natural variability as a function of changing biotic and abiotic factors. To limit the confounding influence of biotic factors, a caging approach with calibrated male organisms from the same population was used. This study applied an in situ deployment at 23 sites of the Rhone basin rivers, complemented by a laboratory experiment assessing the influence of two abiotic factors (temperature and conductivity). The results showed a small effect of conductivity on cellulase activity and a significant effect of temperature on digestive enzyme activity but only at the lowest temperature (7 °C). The experimental conditions allowed us to define an environmental reference value for digestive enzyme activities to select sites where the quality of the water impacted the digestive capacity of the organisms. In addition to the feeding rate, this study showed the relevance of digestive enzymes as biomarkers to be used as an early warning tool to reflect organisms' health and the chemical quality of aquatic ecosystems.

One-year monitoring of core biomarker and digestive enzyme responses in transplanted zebra mussels (Dreissena polymorpha).

A 12-month active biomonitoring study was performed in 2008-2009 on the Vesle river basin (Champagne-Ardenne, France) using the freshwater mussel Dreissena polymorpha as a sentinel species; allochthonous mussels originating from a reference site (Commercy) were exposed at four sites (Bouy, Sept-Saulx, Fismes, Ardre) within the Vesle river basin. Selected core biomarkers (acetylcholinesterase (AChE) activity, glutathione-S transferase (GST) activity, metallothionein concentration), along with digestive enzyme activities (amylase, endocellulase) and energy reserve concentrations (glycogen, lipids), were monitored throughout the study in exposed mussels. At the Fismes and Ardre sites (downstream basin), metallic and organic contamination levels were low but still high enough to elicit AChE and GST activity induction in exposed mussels (chemical stress); besides, chemical pollutants had no apparent deleterious effects on mussel condition. At the Bouy and Sept-Saulx sites (upstream basin), mussels obviously suffered from adverse food conditions which seriously impaired individual physiological state and survival (nutritional stress); food scarcity had however no apparent effects on core biomarker responses. Digestive enzyme activities responded to both chemical and nutritional stresses, the increase in energy outputs (general adaptation syndrome-downstream sites) or the decrease in energy inputs (food scarcity-upstream sites) leading to mid- or long-term induction of digestive carbohydrase activities in exposed mussels (energy optimizing strategy). Complex regulation patterns of these activities require nevertheless the use of a multi-marker approach to allow data interpretation. Besides, their sensitivity to natural confounding environmental factors remains to be precised.

One-year monitoring of reproductive and energy reserve cycles in transplanted zebra mussels (Dreissena polymorpha).

A 12-month active biomonitoring study was performed in 2008-2009 on a northern French river system using the freshwater mussel Dreissena polymorpha as a sentinel species. Allochtonous mussels originating from a reference site (Commercy) were caged at four sites (Bouy, Sept-Saulx, Fismes, Ardre) within the Vesle River basin. The main objective of the study was to characterize the influence of biotic (sex, food availability) and abiotic (temperature, chemicals) factors on the reproductive and energy reserve (glycogen, lipids) cycles of exposed mussels. Both cycles were markedly disturbed at the Bouy and Sept-Saulx sites where the lowest chlorophyll a levels were recorded during the study. At these sites, mussels obviously faced a negative energy balance, as confirmed by the impairment of their physiological state and byssal attachment. At other exposure sites, reproductive and energy reserves cycles were less impacted but were still dependent on the nutritional state of mussels. The latter appeared as a significant natural confounding factor in ecotoxicological survey performed in low polluted areas.

Comprehensive biological effects of a complex field poly-metallic pollution gradient on the New Zealand mudsnail Potamopyrgus antipodarum (Gray).

The Lot River is known to be contaminated by metals, mainly cadmium and zinc, due to a former Zn ore treatment plant in the watershed of the Riou-Mort, a tributary of the Lot River. Many studies have been performed to characterize contamination, but few have assessed its consequences on the biological responses of organisms along the gradient. We exposed adult and juvenile New Zealand freshwater mudsnails Potamopyrgus antipodarum at several sites along the gradient of metal contamination for 28 days. Biological responses were monitored at different levels: individual (survival, growth and fecundity), tissue and biochemical (energy status and vertebrate-like sex steroid levels) to better understand the toxicity mechanisms involved. Accumulation of Cd and Zn was high during exposure. Most of the biological effects observed could be linked to this contamination and were concentration-dependent. Histological lesions of the digestive gland were observed, with hypertrophy of calcium cells and vacuolization of digestive cells. Such effects are likely to explain the decrease of energy status (triglycerides and proteins), juvenile growth and adult fecundity observed at the most polluted site. However the magnitude of the fall in fecundity cannot be attributed only to these tissular effects, indicating another mode of action of Cd or possible confounding factors. Steroid accumulation in snails indicated only organic pollution. Histopathological effects proved the most sensitive endpoint to metal (Cd and Zn) contamination.

Effects of metals on feeding rate and digestive enzymes in Gammarus fossarum: an in situ experiment.

The feeding activity and afterward the assimilation of the products resulting of the food digestion, allow organisms to obtain energy useful for growth, maintenance and reproduction. These biological parameters may be studied to assess the impact of contaminants on the energy metabolism of organisms, which could induce potential effects at an individual level. The studied species was an amphipod Gammarus fossarum, which has a high ecological relevance since it is widespread in European streams and plays a major role in the breakdown of leaf litter. Thus some G. fossarum were transplanted in four sites of a river characterized by metal contamination (Amous River, France). The following parameters were studied: digestive enzymes activities (esterase, beta-glucosidase, beta-galactosidase, amylase and endoglucanase), feeding rate, metal bioaccumulation and survival. Results showed a strong relationship between digestive enzymes activities, feeding rate and metal contents.

Physiological and behavioural responses of Gammarus pulex (Crustacea: Amphipoda) exposed to cadmium.

The aim of this study was to investigate the effects of cadmium on physiological and behavioural responses in Gammarus pulex. In a first experiment, cadmium LC50s for different times were evaluated in 264 h experiment under continuous mode of exposure (LC50(96 h)=82.1 microgL(-1), LC50(120 h)=37.1 microgL(-1), LC50(168 h)=21.6 microgL(-1), LC50(264 h)=10.5 microgL(-1)). In a second experiment, the physiological and behavioural responses of the amphipod exposed to cadmium (0, 7.5 and 15 microgL(-1)) were investigated under laboratory conditions. The mortality and the whole body cadmium concentration of organisms exposed to cadmium were significantly higher than in controls. Concerning physiological responses, cadmium exposure exerted a significant decrease on osmolality and haemolymph Ca(2+) concentration, but not on haemolymph Na(+) and Cl(-) concentrations, whereas the Na(+)/K(+)-ATPase activity was significantly increased. Behavioural responses, such as feeding rate, locomotor and ventilatory activities, were significantly reduced in Cd exposed organisms. Mechanism of cadmium action and consequent energetic reallocation in favour of maintenance functions (i.e., osmoregulation) are discussed. The results of this study indicate that osmolality and locomotor activity in G. pulex could be effective ecophysiological/behavioural markers to monitor freshwater ecosystem and to assess the health of organisms.

Physiological and behavioural responses of Gammarus pulex exposed to acid stress.

Physiological and behavioural responses of the acid-sensitive amphipod Gammarus pulex exposed to a wide range of acid conditions (pH 4.1, 5.1, 6.0) under laboratory conditions were investigated. An exposure of 38 h to acid conditions caused significant decreases in survival rate, osmolality, haemolymph Na+ concentration, ventilatory and locomotor activity compared to organisms exposed to a circumneutral medium (pH 7.9). We highlighted the interest of using individual response distribution, since drastic effects can be detected in organisms exposed to pH 6.0, in particular for osmolality: The response can be divided into two groups, unimpacted and impacted organisms. Moreover this representation permitted to evaluate the health level of individual organisms through the determination of threshold values. Significant correlations between mean pH and mean physiological/behavioural responses were observed. The relationships between individual responses permitted not only to compare endpoints, but also to show that affected organisms were impacted by ionoregulation failure, hypoventilation and low locomotor activity. The energetic reallocation in favour of maintenance functions, such as osmoregulation, is discussed. The results of this study indicate that the values of haemolymph Na+ concentration, osmolality and locomotor activity in G. pulex could be effective ecophysiological/behavioural markers to monitor freshwater ecosystems and to assess the health of organisms or populations.

Measurement of dynamic mobilization of trace metals in sediments using DGT and comparison with bioaccumulation in Chironomus riparius: first results of an experimental study.

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.