Clogging modulates the copper effects on microbial communities of streambed sediments.

The hyporheic zone, i.e. the water-saturated sediment beneath and alongside the riverbed, is exposed to multiple stressors. Agricultural-watershed rivers are frequently exposed to two concomitant stressors: clogging and copper contamination. However, one stressor exposure can increase sensitivity to a second stressor. The aim of this study was to experimentally test the cumulative effects of these two stressors on copper distribution and structural and functional microbial communities responses in the hyporheic zone. A slow filtration column experiment was conducted to compare the effects of 3 treatments of increasing complexity: 'Reference', 'Copper-contaminated' (dissolved copper added at 191 µg L-1), and 'Clogging+Copper' (dissolved copper + addition of 2 cm of fine sediment). Microbial community structure and activities were studied at 4 column sediment depths. The results showed that clogging did not modify the distribution of copper, which remained fixed in the first few centimetres. In the first few centimetres, clogging had a stimulating effect on microbial activities whereas copper had limited effects mainly on leucine aminopeptidase activity and microbial community tolerance to copper. The subsurface zone thus hosts significant different microbial communities from the communities in the deeper zones that were protected from surface stressors. This experiment confirms the valuable filtering role played by the hyporheic zone and shows that microbial responses are strongly correlated to microhabitat-scale physicochemical conditions in sediment.

Improving the design and implementation of sediment fingerprinting studies: summary and outcomes of the TRACING 2021 Scientific School.

Purpose: Identifying best practices for sediment fingerprinting or tracing is important to allow the quantification of sediment contributions from catchment sources. Although sediment fingerprinting has been applied with reasonable success, the deployment of this method remains associated with many issues and limitations. Methods: Seminars and debates were organised during a 4-day Thematic School in October 2021 to come up with concrete suggestions to improve the design and implementation of tracing methods. Results: First, we suggest a better use of geomorphological information to improve study design. Researchers are invited to scrutinise all the knowledge available on the catchment of interest, and to obtain multiple lines of evidence regarding sediment source contributions. Second, we think that scientific knowledge could be improved with local knowledge and we propose a scale of participation describing different levels of involvement of locals in research. Third, we recommend the use of state-of-the-art sediment tracing protocols to conduct sampling, deal with particle size, and examine data before modelling and accounting for the hydro-meteorological context under investigation. Fourth, we promote best practices in modelling, including the importance of running multiple models, selecting appropriate tracers, and reporting on model errors and uncertainty. Fifth, we suggest best practices to share tracing data and samples, which will increase the visibility of the fingerprinting technique in geoscience. Sixth, we suggest that a better formulation of hypotheses could improve our knowledge about erosion and sediment transport processes in a more unified way. Conclusion: With the suggested improvements, sediment fingerprinting, which is interdisciplinary in nature, could play a major role to meet the current and future challenges associated with global change. Supplementary information: The online version contains supplementary material available at 10.1007/s11368-022-03203-1.

Role of Biofilms in Contaminant Bioaccumulation and Trophic Transfer in Aquatic Ecosystems: Current State of Knowledge and Future Challenges.

In freshwater environments, microbial assemblages attached to submerged substrates play an essential role in ecosystem processes such as primary production, supported by periphyton, or organic matter decomposition, supported by microbial communities attached to leaf litter or sediments. These microbial assemblages, also called biofilms, are not only involved in nutrients fluxes but also in contaminants dynamics. Biofilms can accumulate metals and organic contaminants transported by the water flow and/or adsorbed onto substrates. Furthermore, due to their high metabolic activity and their role in aquatic food webs, microbial biofilms are also likely to influence contaminant fate in aquatic ecosystems. In this review, we provide (1) a critical overview of the analytical methods currently in use for detecting and quantifying metals and organic micropollutants in microbial biofilms attached to benthic substrata (rocks, sediments, leaf litter); (2) a review of the distribution of those contaminants within aquatic biofilms and the role of these benthic microbial communities in contaminant fate; (3) a set of future challenges concerning the role of biofilms in contaminant accumulation and trophic transfers in the aquatic food web. This literature review highlighted that most knowledge on the interaction between biofilm and contaminants is focused on contaminants dynamics in periphyton while technical limitations are still preventing a thorough estimation of contaminants accumulation in biofilms attached to leaf litter or sediments. In addition, microbial biofilms represent an important food resource in freshwater ecosystems, yet their role in dietary contaminant exposure has been neglected for a long time, and the importance of biofilms in trophic transfer of contaminants is still understudied.

Relevance of using the non-reactive geochemical signature in sediment core to estimate historical tributary contributions.

Fluvial suspended particulate matter (SPM) fluxes transport large amounts of contaminants that can affect water quality and river ecosystems. To better manage these inputs in river systems, it is essential to identify SPM and sediment sources. Many studies have applied a fingerprinting method based on using metals integrated into a numerical mixing model to estimate source contributions in a watershed. Most fingerprinting studies use contemporary SPM to trace historical inputs, whereas their metal concentrations were modified over time due to anthropogenic inputs. Moreover, total concentrations of these properties are subject to change due to diagenetic processes occurring in stored sediments. The aim of this study was to assess the relevance of using the non-reactive fraction of metals (i.e. metals and metalloids) in fingerprinting studies to estimate the historical contributions of SPM tributary inputs in a sediment core. To assess metal concentrations in the 'conservative' (i.e. non-reactive) fraction, SPM (samples of sources) and sediment core layers (targeted sediments) were subjected to total mineralization and soft extraction, and the non-reactive fraction was obtained by calculating the difference between the two extractions. This approach was applied on a sediment core from the Upper Rhône River (France), using geochemical signature in contemporary SPM of three major tributaries. We showed that the non-reactive fraction retains a higher number of metals in the range test for the deepest layers, which are characterized by significant anthropogenic inputs. Through apportionment modelling using Monte Carlo simulation, we demonstrated that the tributary contributions computed using the non-reactive fraction are more consistent with historical flood and water flow data and have lower uncertainties than with the total fraction. Working with the non-reactive fraction made it possible to decipher historical inputs of SPM using contemporary SPM samples. This approach enables robust identification of sub-catchment areas liable to provide large quantities of SPM. The non-reactive fraction can be used in a variety of environmental conditions and at various spatial and temporal scales to provide a robust quantification of sediment sources.

Acclimation and transgenerational plasticity support increased cadmium tolerance in Gammarus populations exposed to natural metal contamination in headwater streams.

Considering long-term population effects of chronic exposure to contaminants remains limited in ecological risk assessment. Field evidence that multigenerational exposure influences organisms' sensitivity is still scarce, and mechanisms have yet to be elucidated in the environmental context. This study focuses on the crustacean Gammarus fossarum, for which an increased tolerance to cadmium (Cd) has previously been reported in a naturally low-contaminated headwater stream. Our objectives were to investigate whether Cd tolerance is a common phenomenon in headwater populations, and to elucidate the nature of the tolerance and its intergenerational transmission. For this, we carried out an in-depth in situ characterization of Cd exposure (gammarids' caging) and levels of tolerance in nine populations on a regional scale, as well as laboratory maintenance and cross-breeding of contaminated and uncontaminated populations. Acute tolerance levels correlate positively with bioavailable Cd contamination levels among streams. The contaminated and non-contaminated populations differ about two-fold in sensitivity to Cd. Tolerance was found in all age classes of contaminated populations, it can be transiently lost during the year, and it was transmissible to offspring. In addition, tolerance levels dropped significantly when organisms were transferred to a Cd-free environment for two months. These organisms also ceased producing tolerant offspring, confirming a non-genetic transmission of Cd tolerance between generations. These findings support that Cd tolerance corresponds to non-genetic acclimation combined with transgenerational plasticity. Moreover, cross-breeding revealed that tolerance transmission to offspring is not limited to maternal effect. We suggest epigenetics as a plausible mechanism for the plasticity of Cd sensitivity observed in the field. Our results therefore highlight the neglected role of plasticity and non-genetic transmission of modified sensitivities during the long-term exposure of natural populations to environmental contamination.

Water-level fluctuation enhances sediment and trace metal mobility in lake littoral.

Water-level fluctuation (WLF) is a widespread management action in lakes and reservoirs whose impacts on contaminant fate have seldom been investigated. We used near shore hourly measurements (n = 2122) of turbidity (contaminant proxy) and water velocity (sediment resuspension proxy) to track high-frequency contaminant dynamics during a 0.6 m change in water level observed in autumn 2017 in a large French lake. Simultaneously, discrete trace metal measurements highlighted that trapped sediment was more contaminated and finer than surficial sediment supporting that suspended particles (measured by turbidity) were a preferential medium for contaminant mobility. General additive models involving tensor products revealed the enhancement of wind-speed and river discharge effects on turbidity with water draw down. The decrease of the explained deviances by the models over time-lags indicated short time-scale response of turbidity to external forcing. Three of the four major turbid events occurred at the lowest water-level and were concomitant of sediment resuspension as well as precipitation events and/or river flood suggesting a complex interplay among in-lake and watershed processes at controlling sediment mobility during the WLF. These results shed in light that WLF can affect lake littoral hydrodynamic cascading up to the enhancement of contaminant mobility. Sediment resuspension may be an overlooked feature of WLF increasing contamination risk and exposure for littoral organisms with widespread ecological consequences due to the large number of water-level regulated ecosystems.

Dataset of natural metal background levels inferred from pre-industrial palaeochannel sediment cores along the Rhône River (France).

Natural metal background levels in sediments are critical to assess spatial and temporal trends of contamination in hydrosystems and to manage polluted sediments. This is even more sensitive that multi-factors such as geogenic basement, depositional context, and past or long-term pollution can affect the level of metals in sediments. This article provides natural metal background levels and ancillary data (location, chronology, grain-size, total organic carbon - TOC) in pre-industrial sediments along the Rhône River (France). Two distinct areas were selected to take into account the geological variability of the watershed: the Dauphiné Lowlands (Upper Rhône River) and the Tricastin Floodplain (Middle Rhône River). On each area, the sediment cores were retrieved from palaeochannel sequences and the sampled sections were dated by radiocarbon from the Roman to the Modern Times (AD 3-1878). Regulatory metals (Al, Fe, Cd, Cr, Cu, Ni, Pb, and Zn) and other trace elements (Ba, Co, Li, Mg, Mn, Na, P, Sr, Ti, V) were analysed following both Aqua Regia (AR) and Total Extraction (TE) procedures. Classically, TE provides metal concentrations greater than AR because TE includes crystalline lattice, while AR is close to the potentially bio-accessible part of metals (used for ecotoxicological purposes). Due to the small number of samples and to the non-normal distribution of the results, a median-based approach was chosen to establish the geochemical background values and ranges (MGB) for each sample and area. These MGBs are valuable to identify pollution sources, to characterise a contamination (spread and timing), and to estimate the state of rivers regarding pollution legacy. Along the Rhône River, these two continental MGBs were used to reconstruct the metal geo-accumulation trajectories in river sediments from 1965 to 2018 [1].

Partitioning of copper at the confluences of Andean rivers.

The fate of copper (Cu) in rivers impacted by acid drainage remains poorly studied in waters with comparatively low Al and Fe concentrations. This work addresses the role of confluences in controlling the physical and chemical fate of Cu in a system with total molar ratio Cu/Al > 0.2 and Cu/Fe > 0.15. Two consecutive confluences were studied in the upper Mapocho watershed, a densely populated basin with intensive mining located in the Chilean Andes. The inflow had acidic conditions with seasonal variations and Cu up to 9 mg L-1. Lability measurements with diffusive gradient in thin films showed that Cu entered as a dissolved labile form. However, downstream from the confluences a higher pH shifted Cu toward nonlabile compounds and solid phases enriched with Cu. Measurements of x-ray absorption spectroscopy of freshly formed particles showed that composition was dominated by sorbed Cu and Cu(OH)2(s) precipitates, with a higher proportion of sorbed Cu downstream from confluences when pH < 5. Particle size distributions (PSD) measured in field showed that downstream from the confluences the total volume and average diameter of the suspended particles grew progressively, with estimated mean settling velocities increasing from 0.3 to 4.2 cm s-1. As a result, 7-30% of the influent Cu was removed from the river flow. These results highlight that shifts in chemical partition and PSDs in river confluences and the hydrodynamic environments at the river reach level control the mobility of Cu in systems with high Cu/Al and Cu/Fe.

Metal pollution trajectories and mixture risk assessed by combining dated cores and subsurface sediments along a major European river (Rhône River, France).

In European rivers, research and monitoring programmes have targeted metal pollution from bed and floodplain sediments since the mid-20th century by using various sampling and analysis protocols. We propose to characterise metal contamination trajectories since the 1960s based on the joint use of a large amount of data from dated cores and subsurface sediments along the Rhône River (ca. 512 km, Switzerland-France). For the reconstruction of spatio-temporal trends, enrichment factors (EF) and geo-accumulation (Igeo) approaches were compared. The latter index was preferred due to the recurrent lack of grain-size and lithogenic elements in the dataset. Local geochemical backgrounds were established near (1) the Subalps and (2) the Massif Central to consider the geological variability of the watershed. A high contamination (Igeo = 3-5) was found for Cd, Cu and Zn from upstream to downstream over the period 1980-2000. This pattern is consistent with long-term emissions from major cities and the nearby industrial areas of the Upper Rhône (Geneva, Arve Valley), and Middle Rhône (Lyon, Chemical Corridor, Gier Valley). Hotspots due to Cu and Zn leaching from vineyards, mining, and highway runoff were also identified, while Pb was especially driven by industrial sources. The recovery time of pollution in sediment varied according to the metals and was shorter upstream of Lyon (15-20 years) than downstream (30-40 years). More widely, it was faster on the Rhône than along other European rivers (e.g. Seine and Rhine). Finally, the ecotoxicological mixture risk of metal with Persistent Organic Pollutants (POPs) for sediment-dwelling organisms showed a medium "cocktail risk" dominated by metals upstream of Lyon, although it is enhanced due to POPs downstream, and southward to the delta and the Mediterranean Sea. Overall, this study demonstrates the heterogeneity of the contamination trends along large fluvial corridors such as the Rhône River.

Towards simple tools to assess functional effects of contaminants on natural microbial and invertebrate sediment communities.

Surface sediments can accumulate contaminants that affect microorganisms and invertebrates and disturb benthic ecological functions. However, effects of contaminants on ecological functions supported by sediment communities are understudied. Here, we tested the relevance of two simple tools to assess the ecotoxicological effects of metal contamination on natural sediment communities using particulate organic matter breakdown and decomposition as a functional descriptor. To this aim, we performed a 21-day laboratory microcosm experiment to assess the individual and combined effects of Cu and As (nominal concentration of 40 mg kg-1 dw each) using the bait-lamina method (cellulose, bran flakes, and active coal in PVC strips) as well as artificial tablets (cellulose, bran flakes and active coal embedded in an agar matrix). Sediment toxicity was also evaluated using the standardized ostracod toxicity test. Both the bait-lamina and artificial tablet methods showed low effects of As on organic matter breakdown and decomposition but strong effects of Cu on this important ecological function. Both also showed that the presence of Cu and As in mixture in the sediment induced total inhibition of organic matter breakdown and decomposition. The ostracod toxicity test also showed high toxicity of Cu-spiked and Cu-plus-As-spiked sediments and low toxicity of As-spiked sediments. Besides confirming that artificial organic matter substrates are relevant and useful for assessing the functional effects of contaminants on sediment micro- and macro-organism communities, these results suggest that the proposed methods offer promising perspectives for developing tools for use in assessing functional ecotoxicology in the sediment compartment.

Assessment of metal contamination in a small mining- and smelting-affected watershed: high resolution monitoring coupled with spatial analysis by GIS.

The Riou Mort River watershed (SW France), representative of a heavily polluted, small, heterogeneous watershed, represents a major source for the polymetallic pollution of the Lot-Garonne-Gironde fluvial-estuarine system due to former mining and ore-treatment activities. In order to assess spatial distribution of the metal/metalloid contamination in the watershed, a high resolution hydrological and geochemical monitoring were performed during one year at four permanent observation stations. Additionally, thirty-five stream sediment samples were collected at representative key sites and analyzed for metal/metalloid (Cd, Zn, Cu, Pb, As, Sb, Mo, V, Cr, Co, Ni, Th, U and Hg) concentrations. The particulate concentrations in water and stream sediments show high spatial differences for most of the studied elements suggesting strong anthropogenic and/or lithogenic influences; for stream sediments, the sequence of the highest variability, ranging from 100% to 300%, is the following: Mo < Cu < Hg < As < Sb < Cd < Zn < Pb. Multidimensional statistical analyses combined with metal/metalloid maps generated by GIS tool were used to establish relationships between elements, to identify metal/metalloid sources and localize geochemical anomalies attributed to local geochemical background, urban and industrial activities. Finally, this study presents an approach to assess anthropogenic trace metal inputs within this watershed by combining lithology-dependent geochemical background values, metal/metalloid concentrations in stream sediments and mass balances of element fluxes at four key sites. The strongest anthropogenic contributions to particulate element fluxes are 90-95% for Cd, Zn and Hg in downstream sub-catchments. The localisation of anthropogenic metal/metalloid sources in restricted areas offers a great opportunity to further significantly reduce metal emissions and restore the Lot-Garonne-Gironde fluvial-estuarine ecosystem.

Experimental Warming Differentially Influences the Vulnerability of Phototrophic and Heterotrophic Periphytic Communities to Copper Toxicity.

Aquatic ecosystems are generally subjected to multiple perturbations due to simultaneous or successive combinations of various natural and anthropogenic environmental pressures. To better assess and predict the resulting ecological consequences, increasing attention should be given to the accumulation of stresses on freshwater ecosystems and its effects on the vulnerability of aquatic organisms, including microbial communities, which play crucial functional roles. Here we used a microcosm study to assess the influence of an experimental warming on the vulnerability of phototrophic and heterotrophic periphytic communities to acute and chronic copper (Cu) toxicity. Natural periphytic communities were submitted for 4 weeks to three different temperatures (18, 23, and 28°C) in microcosms contaminated (at about 15 µg L-1) or not with Cu. The vulnerability of both phototrophic and heterotrophic microbial communities to subsequent acute Cu stress was then assessed by measuring their levels of sensitivity to Cu from bioassays targeting phototrophic (photosynthetic activity) and heterotrophic (ß-glucosidase and leucine aminopeptidase extracellular enzymatic activities) microbial functions. We postulated that both the increase in temperature and the chronic Cu exposure would modify microbial community structure, thus leading to changes in the capacity of phototrophic and heterotrophic communities to tolerate subsequent acute exposure to Cu. Our results demonstrated that the influence of temperature on the vulnerability of phototrophic and heterotrophic microbial communities to Cu toxicity can vary greatly according to function studied. These findings emphasize the importance of considering different functional compartments and different functional descriptors to better assess the vulnerability of periphyton to multiple stresses and predict the risks induced by multiple stressors for ecosystem balance and functioning.

Environmental Concentrations of Copper, Alone or in Mixture With Arsenic, Can Impact River Sediment Microbial Community Structure and Functions.

In many aquatic ecosystems, sediments are an essential compartment, which supports high levels of specific and functional biodiversity thus contributing to ecological functioning. Sediments are exposed to inputs from ground or surface waters and from surrounding watershed that can lead to the accumulation of toxic and persistent contaminants potentially harmful for benthic sediment-living communities, including microbial assemblages. As benthic microbial communities play crucial roles in ecological processes such as organic matter recycling and biomass production, we performed a 21-day laboratory channel experiment to assess the structural and functional impact of metals on natural microbial communities chronically exposed to sediments spiked with copper (Cu) and/or arsenic (As) alone or mixed at environmentally relevant concentrations (40 mg kg-1 for each metal). Heterotrophic microbial community responses to metals were evaluated both in terms of genetic structure (using ARISA analysis) and functional potential (using exoenzymatic, metabolic and functional genes analyses). Exposure to Cu had rapid marked effects on the structure and most of the functions of the exposed communities. Exposure to As had almost undetectable effects, possibly due to both lack of As bioavailability or toxicity toward the exposed communities. However, when the two metals were combined, certain functional responses suggested a possible interaction between Cu and As toxicity on heterotrophic communities. We also observed temporal dynamics in the functional response of sediment communities to chronic Cu exposure, alone or in mixture, with some functions being resilient and others being impacted throughout the experiment or only after several weeks of exposure. Taken together, these findings reveal that metal contamination of sediment could impact both the genetic structure and the functional potential of chronically exposed microbial communities. Given their functional role in aquatic ecosystems, it poses an ecological risk as it may impact ecosystem functioning.

Looking at biological community level to improve ecotoxicological assessment of freshwater sediments: report on a first French-Swiss workshop.

The first French-Swiss workshop on ecotoxicology of freshwater sediment communities was co-organized by the French Research Institute of Science and Technology for Environment and Agriculture (Irstea) and the Swiss Centre for Applied Ecotoxicology (Ecotox Centre EAWAG-EPFL) in Villié-Morgon (Beaujolais Region, France) on April 27-28, 2017. The workshop brought together scientists working in different fields of expertise (ecotoxicologists, ecologists, environmental chemists…), environmental stakeholder groups and managers, as well as economic players (start-ups and consultancies) to better connect research needs of potential end-users with research outputs. The objectives of this workshop were (i) to establish the state of the art of research in the characterization of sediment contamination and in the evaluation of the effects on sediment-associated biological communities and ecosystem functioning and (ii) to give an overview of the French and Swiss regulations dealing with the assessment of contaminated sediments in freshwater ecosystems. The ultimate goal was to collectively identify research needs and knowledge gaps, as well as to highlight ways to improve the ecotoxicological assessment of sediments in freshwater environments by further considering the structure and functions of associated microbial and invertebrate communities.

Groundwater contributions to metal transport in a small river affected by mining and smelting waste.

The Riou Mort watershed, strongly affected by former coal mining and Zn ore treatment, has been the major source of the historical polymetallic pollution of the Lot-Garonne-Gironde fluvial-estuarine system. Two decades after the end of ore treatment, the former industrial area still contributes important amounts of metals/metalloids from various, partly unidentified, sources to the downstream river system. This study presents the high spatial variability of metal/metalloid (Cd, Zn, As, Sb, U, V) concentrations in water and suspended particulate matter (SPM) from eight observation sites during a short, intense flood event. Despite important dilution effects, the observed concentration levels at the different sites suggested additional Cd and Zn inputs, probably from polluted groundwater. This formerly unknown metal source was then localized and characterized by sampling water and SPM along two longitudinal profiles during different hydrological situations. Groundwater inputs of "truly dissolved" (<0.02 microm) Cd and Zn occurred along approximately 200 m, contributing 43% and 28% to the total annual (2004) Cd and Zn fluxes in the Riou Mort River. The estimated groundwater concentrations of Cd and Zn (2500-6700 and 83,000-170,000 microg l(-1), respectively) in the source zone were consistent with values measured in samples from the near aquifer (5400-13,000 and 200,000-400,000 microg l(-1)). The present work induced concrete remediation actions (pumping and treatment of the polluted groundwater), that are expected to strongly reduce dissolved Cd and Zn emissions into the Riou Mort River.

Use of Gammarus fossarum (Amphipoda) embryo for toxicity testing: A case study with cadmium.

The effects of environmental contaminants on arthropod embryo stages have been poorly investigated in ecotoxicology. Moreover, many of these tests used hatching success as the sole metric, although it is possible to detect many more subtle effects. After a detailed description of embryogenesis in Gammarus fossarum, the present study reports on the sublethal effects of cadmium (Cd) exposure during embryonic development in G. fossarum. Embryos were first directly exposed in multiwell plates throughout the entire embryonic cycle (23 d) to increasing Cd concentrations (0, 1.5, and 3.0 µg/L; 120 embryos/concentration). Then, to assess the representativeness of the gammarid embryo assay performed in multiwell plates, embryos were exposed to similar Cd concentrations through the maternal open brood pouch. Next, to pinpoint sensitive periods of development, embryos were directly exposed to 3.0 µg/L of Cd for shorter periods of time: during gastrulation, organogenesis, and hatching. After hatching, the following parameters were measured in the newborn individuals: 1) body mass; 2) activity of the enzyme phenoloxidase, a key enzyme of the arthropod immune system; and 3) locomotor activity. Phenoloxidase activity was strongly inhibited in newborn individuals of embryos exposed (either in multiwell plates or in the maternal brood pouch) to 3.0 µg/L Cd throughout embryonic development. Furthermore, strong detrimental locomotor effects were observed in newborn individuals of embryos directly exposed to 3.0 µg/L. Exposures for shorter periods of time were not sufficient to induce such effects; no sensitive period could be determined. By bringing new insights into a critical time window of exposure, the gammarid embryo assay could provide a novel and interesting addition to existing bioassays in gammarids. Environ Toxicol Chem 2017;36:2436-2443. © 2017 SETAC.

Influence of temperature in pollution-induced community tolerance approaches used to assess effects of copper on freshwater phototrophic periphyton.

By measuring levels of tolerance to toxicants in microbial communities using functional toxicity tests under controlled conditions, pollution-induced community tolerance (PICT) approaches offer an effect-based tool to assess the ecological risk of chemicals in aquatic systems. However, induced tolerance of exposed microbial communities cannot always be attributed solely to the presence of toxicants as various environmental factors, such as temperature, can also be involved. Several PICT studies have been conducted to assess the effects of copper (Cu) on phototrophic periphyton, but little is known about the influence of temperature on the response of these microbial communities to acute and chronic exposure to Cu. Here, we report on a microcosm approach to assess the effects of two contrasting temperatures (18°C and 28°C) on (i) the baseline level of Cu tolerance in non-Cu-exposed phototrophic periphyton (i.e. effect of temperature on tolerance baseline), (ii) Cu tolerance acquisition by phototrophic periphyton in response to a 3-week chronic exposure to Cu at a nominal concentration of 60µgL-1 (i.e. effect of temperature on PICT selection) and (iii) tolerance measured during short-term toxicity tests (i.e. effect of temperature on PICT detection). The aim was to evaluate how temperature conditions during the different phases of the PICT approaches may modify the causal relationship between chronic Cu exposure and measured Cu tolerance levels. Our results evidence the influence of temperature both on the basal capacity of phototrophic periphyton to tolerate subsequent exposure to Cu (i.e. influence on tolerance baseline) and on its capacity to acquire tolerance following chronic exposure to Cu (i.e. influence on PICT selection). Hence temperature must be considered when using PICT to establish causal links between chronic Cu exposure and effects on phototrophic periphyton.

Changes in copper toxicity towards diatom communities with experimental warming.

Biological communities in aquatic environments most commonly face multiple stress, where natural and anthropogenic stressors often act jointly. Their interactions are most easily assessed using short cycle organisms such as periphytic diatoms. In this experiment, we analyzed the combined effects of copper exposure and warming on diatom successions over 6 weeks. Natural biofilm collected in winter was left to grow in mesocosms exposed or unexposed to realistic Cu concentrations at four different temperatures. Separate and joint impacts of the two stressors were determined through structural and functional endpoints. Both temperature and copper influenced the biological responses; their interaction, when significant, was always antagonistic. Diatom communities gradually changed with rising temperature. Under copper exposure, the dominant Planothidium lanceolatum was superseded by Achnanthidium exiguum, which accounted for about 70% relative abundance in the warmest conditions (18-23°C). Tolerance to copper was derived from dose-response curves based on photosynthesis inhibition. Cu-induced community tolerance was always found, but it decreased with warming and time. Biodiversity loss associated with lower Cu tolerance under combined Cu exposure and increasing temperatures evidences the major influence of cumulative stressors on aquatic health. These results highlight the crucial interplay between environmental stressors, which are expected to intensify with climate change.

Temperature modulates phototrophic periphyton response to chronic copper exposure.

Streams located in vineyard areas are highly prone to metal pollution. In a context of global change, aquatic systems are generally subjected to multi-stress conditions due to multiple chemical and/or physical pressures. Among various environmental factors that modulate the ecological effects of toxicants, special attention should be paid to climate change, which is driving an increase in extreme climate events such as sharp temperature rises. In lotic ecosystems, periphyton ensures key ecological functions such as primary production and nutrient cycling. However, although the effects of metals on microbial communities are relatively well known, there is scant data on possible interactions between temperature increase and metal pollution. Here we led a study to evaluate the influence of temperature on the response of phototrophic periphyton to copper (Cu) exposure. Winter communities, collected in a 8 °C river water, were subjected for six weeks to four thermal conditions in microcosms in presence or not of Cu (nominal concentration of 15 µg L(-1)). At the initial river temperature (8 °C), our results confirmed the chronic impact of Cu on periphyton, both in terms of structure (biomass, distribution of algal groups, diatomic composition) and function (photosynthetic efficiency). At higher temperatures (13, 18 and 23 °C), Cu effects were modulated. Indeed, temperature increase reduced Cu effects on algal biomass, algal class proportions, diatom assemblage composition and photosynthetic efficiency. This reduction of Cu effects on periphyton may be related to lower bioaccumulation of Cu and/or to selection of more Cu-tolerant species at higher temperatures.

Improved short-term toxicity test protocol to assess metal tolerance in phototrophic periphyton: toward standardization of PICT approaches.

Pollution-induced community tolerance (PICT) approaches involve comparing tolerance levels of natural communities to a particular contaminant or a contaminant mixture using short-term toxicity tests performed under controlled conditions. However, results from toxicity tests can be modulated by various environmental and experimental conditions, raising questions about their reproducibility and comparability. In this context, the present study aimed to determine the influence of exposure duration, periphyton suspension concentration, and periphyton maturation stage on the measurement of short-term effects of copper on phototrophic periphyton communities. Our results showed the very weak influence of exposure duration in the tested range (2-6 h) on toxicity level, whereas periphyton biomass in the tested suspension (in terms of both chlorophyll a concentrations and dry weight), proved a crucial determinant in toxicity assessment. Results also highlighted the potential tolerance increase with the periphyton maturation stage. This parameter conditioned the positive linear relationship between tolerance level and periphyton suspension concentration, leading to an increase in the linear regression slope with the maturation stage. This suggests that such a relationship is probably highly periphyton-dependent. Consequently, to enable data toxicity comparisons, an a priori normalization of the periphyton suspension biomass is necessary, and PICT approaches require the use, as much of possible, of periphyton with similar maturation stage. Finally, the present study clearly shows that a better standardization of PICT approaches could help to improve reproducibility. It could thus facilitate the comparison of tolerance levels measured in the same study (e.g., spatial and/or temporal and/or inter-treatment comparison) as well as the comparison obtained from different experimental and in situ research.