Experimental testing of two urban stressors on freshwater biofilms.

Aquatic ecosystems and their communities are exposed to numerous stressors of various natures (chemical and physical), whose impacts are often poorly documented. In urban areas, the use of biocides such as dodecyldimethylbenzylammonium chloride (DDBAC) and their subsequent release in wastewater result in their transfer to urban aquatic ecosystems. DDBAC is known to be toxic to most aquatic organisms. Artificial light at night (ALAN) is another stressor that is increasing globally, especially in urban areas. ALAN may have a negative impact on photosynthetic cycles of periphytic biofilms, which in turn may result in changes in their metabolic functioning. Moreover, studies suggest that exposure to artificial light could increase the biocidal effect of DDBAC on biofilms. The present study investigates the individual and combined effects of DDBAC and/or ALAN on the functioning and structure of photosynthetic biofilms. We exposed biofilms in artificial channels to a nominal concentration of 30 mg.L-1 of DDBAC and/or ALAN for 10 days. ALAN modified DDBAC exposure, decreasing concentrations in the water but not accumulation in biofilms. DDBAC had negative impacts on biofilm functioning and structure. Photosynthetic activity was inhibited by > 90% after 2 days of exposure, compared to the controls, and did not recover over the duration of the experiment. Biofilm composition was also impacted, with a marked decrease in green algae and the disappearance of microfauna under DDBAC exposure. The integrity of algal cells was compromised where DDBAC exposure altered the chloroplasts and chlorophyll content. Impacts on autotrophs were also observed through a shift in lipid profiles, in particular a strong decrease in glycolipid content was noted. We found no significant interactive effect of ALAN and DDBAC on the studied endpoints.

Influence of a hydroelectric dam on fish mercury contamination along the Sinnamary River (French Guiana).

The Petit Saut hydroelectric dam and the upstream and downstream areas of the Sinnamary River in French Guiana (Amazon basin) have been studied from 1993 to 2020. The nearly thirty-years-long study of the monitoring of total mercury concentration in fish and the physicochemical survey of the environment made it possible to demonstrate the impact of the flooding of the forest and the role of the hydroelectric dam on the methylation of mercury. Results show that the physicochemical modifications generated by the construction of the dam led to a significant production of methylmercury (MeHg) in the anoxic part of the reservoir and downstream of the river leading to a strong spatio-temporal impact of the dam. Seven species of fishes are studied and their mercury concentrations vary according to many parameters: fish diet, position in the water column, site, lake oxycline level and time.

Toxicity assessment of environmental MPs and NPs and polystyrene NPs on the bivalve Corbicula fluminea using a multi-marker approach.

Small plastic particles, microplastics (MPs) and nanoplastics (NPs) represent a major threat in aquatic environments. Freshwater organisms are exposed to MPs and NPs, particularly in industrial and urban areas. The present study aimed to compare the toxicity between polystyrene NPs (PS NPs) and environmental microplastics (ENV MPs) and nanoplastics (ENV NPs) generated from macro-sized debris collected in the Garonne River on the freshwater bivalve C. fluminea. The organisms were exposed to the different plastic particles at three environmentally relevant concentrations: 0.008, 10, and 100 µg L-1 for 21 days. The biological responses of organisms were assessed using a multi-biomarker approach from the sub-individual to the individual level. The results demonstrated that: i) ENV NPs triggered more effects on detoxification processes and immune response, confirming that using manufactured NPs for laboratory exposure can lead to misleading conclusions on the risks posed by plastic particles; ii) effects of ENV MPs were less marked than ENV NPs, emphasizing the importance of testing a size continuum of plastic particles from NPs to MPs; iii) some effects were only observed for the low and/or intermediate concentrations tested, underlining the importance of using environmentally relevant concentrations. In light of these results, laboratory studies should be continued by exposing aquatic species to environmental MPs and NPs. The properties of these particles have to be characterized for a better risk assessment of environmental plastic particles.

Impact of nickel mining in New Caledonia on marbled eels Anguilla marmorata.

New Caledonia is particularly affected by nickel open pit mining activities because of the presence of ultramafic soils rich in metals. The particles dispersed by atmospheric transport and soil erosion during the excavation of nickel end up by deposition or leaching in rivers where they may be bioaccumulated by organisms living downstream the mines. Despite alarming freshwater metals concentrations, no study investigated the level of their bioaccumulation in eels, and if high bioaccumulation levels occur, the potential consequences on their health. The aim of this study was to determine how eels Anguilla marmorata are impacted in situ by metals issued from mining activity by measuring: morphometric parameters; metal concentrations in tissues and organs and transcription levels of target genes encoding proteins involved in several metabolic key functions. Among organs, liver was found to be the most affected by mining with average nickel concentrations of 5.14 mg/kg versus 1.63 mg/kg for eels away from mines leading to dysregulation of numerous genes involved in oxidative stress, DNA repair, apoptosis, reproduction and both lipid and mitochondrial metabolisms. This study should allow us to define in an integrated way if metals released by mining activities influence metals bioaccumulation in eels and induce biological effects.

Transfer and Transcriptomic Profiling in Liver and Brain of European Eels (Anguilla anguilla) After Diet-borne Exposure to Gold Nanoparticles.

A nanometric revolution is underway, promising technical innovations in a wide range of applications and leading to a potential boost in environmental discharges. The propensity of nanoparticles (NPs) to be transferred throughout trophic chains and to generate toxicity was mainly assessed in primary consumers, whereas a lack of knowledge for higher trophic levels persists. The present study focused on a predatory fish, the European eel (Anguilla anguilla) exposed to gold NPs (AuNPs; 10 nm, polyethylene glycol-coated) for 21 d at 3 concentration levels in food: 0 (NP0), 1 (NP1), and 10 (NP10) mg Au kg-1 . Transfer was assessed by Au quantification in eel tissues, and transcriptomic responses in the liver and brain were revealed by a high-throughput RNA-sequencing approach. Eels fed at NP10 presented an erratic feeding behavior, whereas Au quantification only indicated transfer to intestine and kidney of NP1-exposed eels. Sequencing of RNA was performed in NP0 and NP1 eels. A total of 258 genes and 156 genes were significantly differentially transcribed in response to AuNP trophic exposure in the liver and brain, respectively. Enrichment analysis highlighted modifications in the immune system-related processes in the liver. In addition, results pointed out a shared response of both organs regarding 13 genes, most of them being involved in immune functions. This finding may shed light on the mode of action and toxicity of AuNPs in fish. Environ Toxicol Chem 2020;39:2450-2461. © 2020 SETAC.

Ecotoxicity of polyethylene nanoplastics from the North Atlantic oceanic gyre on freshwater and marine organisms (microalgae and filter-feeding bivalves).

Each year, 5 to 10 million tons of plastic waste is dumped in the oceans via freshwaters and accumulated in huge oceanic gyres. Under the effect of several abiotic factors, macro plastic wastes (or plastic wastes with macro sizes) are fractionated into microplastics (MP) and finally reach the nanometric size (nanoplastic NP). To reveal potential toxic impacts of these NPs, two microalgae, Scenedemus subspicatus (freshwater green algae), and Thalassiosira weissiflogii (marine diatom) were exposed for up to 48 h at 1, 10, 100, 1000, and 10,000 µg/L to reference polyethylene NPs (PER) or NPs made from polyethylene collected in the North Atlantic gyre (PEN, 7th continent expedition in 2015). Freshwater filter-feeding bivalves, Corbicula fluminea, were exposed to 1000 µg/L of PER and PEN for 48 h to study a possible modification of their filtration or digestion capacity. The results show that PER and PEN do not influence the cell growth of T. weissiflogii, but the PEN exposure causes growth inhibition of S. subspicatus for all exposure concentrations tested. This growth inhibition is enhanced for a higher concentration of PER or PEN (10,000 µg/L) in S. subspicatus. The marine diatom T. weissiflogii appears to be less impacted by plastic pollution than the green algae S. subspicatus for the exposure time. Exposure to NPs does not lead to any alteration of bivalve filtration; however, fecal and pseudo-fecal production increased after PEN exposure, suggesting the implementation of rejection mechanisms for inedible particles.

Mercury contamination levels in the bioindicator piscivorous fish Hoplias aïmara in French Guiana rivers: mapping for risk assessment.

In French Guiana, native populations present high level of mercury contamination, which has been linked to the consumption of contaminated fishes. The goal of this study is to undertake a cartography of mercury contamination levels in fishes from the six main Guiana rivers. The selected species for this study is the ubiquitous piscivorous fish Hoplias aimara. A total number of 575 fishes from 134 discrete fishing sites are regrouped into 51 river sectors. Results from this study permits to rank the six main Guiana rivers by their mean level of contamination: Oyapock (0.548 mg kg-1), Comté (0.624 mg kg-1), Maroni (0.671 mg kg-1), Approuague (0.684 mg kg-1), Mana (0.675 mg kg-1), and Sinnamary (1.025 mg kg-1). The contamination is however not spatially homogenous along each river, and a map of the different levels of mercury contamination in fishes is provided. Sectors of low mean Hg contamination are observed both upstream (0.471 mg kg-1) and downstream (0.424 mg kg-1), corresponding to areas without any influence of gold mining activities and areas under the influence of estuarine dilution, respectively. Anoxia and gold mining activities are found to be the two main factors responsible for the high mercury concentration in fish muscles. While mean levels of mercury contaminations are higher in anoxia areas (1.029 mg kg-1), contaminations induced by gold mining activities (0.717 mg kg-1) present the most harmful consequences to human populations. No significant differences in Hg concentrations are observed between 2005 and 2014 for neither a pristine nor a gold mining area, while Hg concentration differences are observed between former (0.550 mg kg-1) and current gold mining sites (0.717 mg kg-1).

Impact of diuron and S-metolachlor on the freshwater diatom Gomphonema gracile: Complementarity between fatty acid profiles and different kinds of ecotoxicological impact-endpoints.

Fatty acids (FA) are crucial for the maintenance of membrane fluidity and play a central role in metabolic energy storage. Polyunsaturated fatty acids play an essential ecological role since they are key parameters in the nutritional value of algae. Pesticide impacts on fatty acid profiles have been documented in marine microalgae, but remain understudied in freshwater diatoms. The aims of this study were to: 1) investigate the impact of diuron and S-metolachlor on "classical descriptors" (photosynthesis, growth rate, pigment contents, and on the expression levels of target genes in freshwater diatoms), 2) examine the impact of these pesticides on diatom fatty acid profiles and finally, 3) compare fatty acid profiles and "classical descriptor" responses in order to evaluate their complementarity and ecological role. To address this issue, the model freshwater diatom Gomphonema gracile was exposed during seven days to diuron and S-metolachlor at 10 µg.L-1. G. gracile was mostly composed of the following fatty acids: 20:5n3; 16:1; 16:0; 16:3n4; 14:0 and 20:4n6 and highly unsaturated fatty acids were overall the best represented fatty acid class. S-metolachlor decreased the growth rate and chlorophyll a content of G. gracile and induced the expression of cox1, nad5, d1 and cat genes, while no significant impacts were observed on photosynthesis and carotenoid content. In a more global way, S-metolachlor did not impact the fatty acid profiles of G. gracile. Diuron inhibited photosynthesis, growth rate, chlorophyll a content and induced cat and d1 gene expressions but no significant effect was observed on carotenoid content. Diuron decreased the percentage of highly unsaturated fatty acids but increased the percentage of monounsaturated fatty acids. These results demonstrated that fatty acids responded to diuron conversely to pigment content, suggesting that fatty acids can inform on energy content variation in diatoms subjected to herbicide stress.

Trophic transfer and effects of gold nanoparticles (AuNPs) in Gammarus fossarum from contaminated periphytic biofilm.

This work addressed the trophic transfer and effects of functionalized gold nanoparticles (AuNPs) from periphytic biofilms to the crustacean Gammarus fossarum. Biofilms were exposed for 48 h to 10 nm positively charged functionalized AuNPs at two concentrations, 4.6 and 46 mg/L, and crustaceans G. fossarum grazed on these for 7 days, with daily biofilm renewal. Gold bioaccumulation in biofilm and crustacean were measured to estimate the trophic transfer ratio of these AuNP, and, for the first time, a transcriptomic approach and transmission electron microscopy observations in the crustacean were made. These two approaches showed cellular damage caused by oxidative stress and, in particular, an impact of these AuNPs on mitochondrial respiration. Modulation of digestive enzyme activity was also observed, suggesting modifications of digestive functions. The damage due to these nanoparticles could then have vital consequences for the organisms during chronic exposure.

Can pesticides, copper and seasonal water temperature explain the seagrass Zostera noltei decline in the Arcachon bay?

Dwarf eelgrasses (Zostera noltei) populations have decreased since 2005 in Arcachon Bay (southwest France). Various stressors have been pointed out, however the role of xenobiotics like pesticides or copper (Cu) and of parameters like water temperature warming have not yet been explored. To determine their impact, Z. noltei individuals were collected in a pollution-free site and transferred to the laboratory in seawater microcosms. This dwarf eelgrass was exposed to a pesticide cocktail and copper, alone or simultaneously, at temperatures (10°C, 20°C, 28°C) representative of different seasons. After a two-week contamination, leaf growth, leaf bioaccumulation of Cu, and differential expression of target genes were studied. Eelgrasses bioaccumulated Cu regardless of the temperature, with reduced efficiency in the presence of the Cu and pesticide cocktail at the two higher temperatures. High temperature also exacerbated the effect of contaminants, leading to growth inhibition and differential gene expression. Mitochondrial activity was strongly impacted and higher mortality rates occurred. Experimental results have been confirmed during field survey. This is the first report on the impacts on Z. noltei of pesticides and Cu associate to temperature.

Biosynthesis of gold nanoparticles by the living freshwater diatom Eolimna minima, a species developed in river biofilms.

Testing biotransformation capacities of living aquatic microalgae diatoms to naturally synthetize gold nanoparticles (AuNP) from gold salts and assessing aftereffects on their viability by microscope observations is a great challenge. In this work, a laboratory experiment was conducted, which aimed to observe (i) directly by transmission electronic and light microscopy and (ii) through indirect measurements (UV-visible spectroscopy) the periphytic freshwater diatom Eolimna minima exposed to gold salts. This work revealed the capacity of E. minima to intracellularly biosynthetize AuNP and to tolerate it. AuNP synthesis appears as a mechanism of detoxification to protect diatom from gold salt contamination. We also pointed out the risks associated with the spread of diatoms full of AuNP, through the trophic web of freshwater ecosystems. The preponderant part of the diatoms in natural biofilms associated with their position at the basis of the trophic webs in rivers could then make them responsible for the contamination of their consumers (grazer animals) and consequently for the potential release of AuNP through the entire food web.

Herbicide toxicity on river biofilms assessed by pulse amplitude modulated (PAM) fluorometry.

The use of Rapid light curves (RLCs) as a toxicity endpoint for river biofilms was examined in this study and compared to "classical fluorescence parameters" i.e. minimal fluorescence (F0), optimal and effective quantum yields of photosystem II (Fv/Fm and ФPSII). Measurements were performed after exposure to five concentrations of diuron (from 0.3 to 33.4µgL(-1)), its main degradation product (DCPMU) (from 1.0 to 1014µgL(-1)) and norflurazon (from 0.6 to 585µgL(-1)) with the lowest exposure concentrations corresponding to levels regularly encountered in chronically contaminated sites. Biofilm responses were evaluated after 1, 5, 7 and 14 days of exposure to the different toxicants. Overall, the responses of both "classical fluorescence parameters" and RLC endpoints were highly time dependent and related to the mode of action of the different compounds. Interestingly, parameters calculated from RLCs (α, ETRmax and Ik) were useful early markers of pesticide exposure since they revealed significant effects of all the tested toxicants from the first day of exposure. In comparison, classical fluorescence endpoints (F0 and Fv/Fm) measured at day 1 were only affected in the DCPMU treatment. Our results demonstrated the interest of RLCs as early markers of toxicant exposure particularly when working with toxicants with less specific mode of action than PSII inhibitors.

Is the toxicity of pesticide mixtures on river biofilm accounted for solely by the major compounds identified?

Comparative effects of long-term exposure to Polar Organic Chemical Integrative Sampler (POCIS) extracts (PE) and to a reconstituted mixture based on the major compounds quantified in the PE were evaluated on river biofilm communities. The study aimed to characterize the effects of long-term and low-dose exposure to pesticides on natural biofilm communities and to evaluate if the effects due to PE exposure could be explained solely by the major compounds identified in the extracts. Biofilms from an uncontaminated site were exposed in artificial channels to realistic environmental concentrations using diluted PE, with the 12 major compounds quantified in the extracts (Mix) or with water not containing pesticides (Ctr). Significant differences between biofilms exposed to pesticides or not were observed with regard to diatom density, biomass (dry weight and ash-free dry mass), photosynthetic efficiency (ΦpsII) and antioxidant enzyme activities. After 14 days of exposure to the different treatments, the observed trend towards a decrease of mean diatom cell biovolumes in samples exposed to pesticides was related to the control biofilms' higher relative abundance of large species like Cocconeis placentula or Amphora copulata and lower relative abundance of small species like Eolimna minima compared to the contaminated ones. Principal component analyses clearly separated contaminated (PE and Mix) from non-contaminated (Ctr) biofilms; on the contrary, the analyses did not reveal separation between biofilms exposed to PE or to the 12 major compounds identified in the extract.

Single and mixture effects of pesticides and a degradation product on fluvial biofilms.

The Morcille River located in the Beaujolais vineyard area (Eastern France) is subjected to strong vine-growing pressure leading to the contamination by a range of herbicides and fungicides of the surrounding freshwater environment. Particularly high concentrations of norflurazon, desmethyl norflurazon and tebuconazole were recorded in spring 2010 at the downstream site of the river. Despite their occurrence in rivers, scarce toxicity data are available for these products, in particular in the case of desmethyl norflurazon (main norflurazon degradation product). Furthermore, the toxicity data are generally available only for single compounds and are issued from single species toxicity tests, leading to a lack of ecological relevance. Consequently, this study was undertaken to evaluate the toxic effects of norflurazon, desmethyl norflurazon and tebuconazole singly and in a ternary mixture on fluvial biofilm. Toxicity tests were performed in microplates for 48 h. Photosynthetic endpoints were measured using pulse amplitude-modulated fluorometry; diatom densities and taxonomic composition were determined. After 48 h of exposure, significant effects on optimal quantum yield (F v/F m) for desmethyl norflurazon and mixture were observed.

Environmental effects of realistic pesticide mixtures on natural biofilm communities with different exposure histories.

This study deals with the use of Polar Organic Chemical Integrative Sampler (POCIS) extracts to assess the impact of low-dose pesticide mixtures on natural biofilm communities originating from either a chronically contaminated or a reference field site. To investigate how natural biofilm communities, pre-exposed to pesticides in situ or not might respond to environmentally realistic changes in pesticide pressure, they were exposed to either clean water or to POCIS extracts (PE) in order to represent toxic pressure with a realistic pesticide mixture directly isolated from the field. The impacts of PE were assessed on structure, physiology and growth of biofilms. Initial levels of tolerance of phototrophic communities to PE were also estimated at day 0. PE exposure led to negative effects on diatom growth kinetics independently of in-field biofilm exposure history. In contrast, the impacts observed on dry weight, ash-free dry mass and algal fluorescence-related parameters followed different trends depending on biofilm origin. Exposure to PE induced changes in diatom assemblages for the biofilm originating from the reference field site with higher relative abundance of Eolimna minima and Nitzschia palea with PE exposure. Initial tolerance of phototrophic communities to PE was 8-fold higher for the biofilm originating from the chronically contaminated site compared to the reference field site. The use of POCIS extracts allowed us to highlight both chronic impacts of low doses of a mixture of pesticides on natural communities with regard to biofilm exposure history as well as initial levels of tolerance of phototrophic communities.

Cadmium decontamination and reversal potential of teratological forms of the diatom Planothidium frequentissimum (Bacillariophyceae) after experimental contamination.

While the induction of teratology by cadmium (Cd) on diatoms is already known, reversal kinetics are not well documented. This study aims to understand the viability of diatoms exhibiting teratological frustules and their reproduction capacities within a Cd-impacted population to predict their return to normal diatom forms. We worked on a frequently encountered species in French hydrosystems: Planothidium frequentissimum (Lange-Bertalot) Round & L. Bukhtiyarova. First, a 21-d contamination phase highlighted increasing inductionof different teratological types in response to two levels of Cd contamination: 20 and 100 µg · L(-1) . The deformity counting indicated that Cd firstly generated striae and mixed teratologies, then affected the central area and the valves. Second, a 28-d decontamination phase demonstrated the Cd depuration capacity of Planothidium frequentissimum. Cd half-lives appeared relatively low, ~6 d for the 100 µg · L(-1) condition. Moreover, the decontamination phase showed a decrease in teratology abundances, but a still incomplete recovery after 28 d. Deformations of the striae appeared to be the most sustainable phenotype since they were still significantly higher than in reference cultures at the end of the decontamination phase for both Cd cultures.

Recovery potential of periphytic biofilms translocated in artificial streams after industrial contamination (Cd and Zn).

Metal wastes can significantly disturb aquatic communities, particularly photosynthetic organisms, the main primary producers in freshwater running ecosystems. In this study, biofilms and diatoms were used as bioindicators to characterize the kinetics of biofilm recovery. An experimental decontamination study was conducted under laboratory conditions, after biofilm colonisation at a site subject to discharge of industrial metals (Zn and Cd) and in parallel at an upstream site, metal-free, considered as a control. After 24 days of colonisation, biofilms were translocated and maintained in the laboratory for 56 days under clean conditions (control and decontamination) or metal contamination. Various tests were conducted from the community level--measures of metal bioaccumulation, cell densities and taxonomic investigations, to the individual level--measures of teratological forms. After 56 days of decontamination, Zn and Cd concentrations in decontaminated biofilms showed a sharp decline, respectively ranging from 6.7 ± 2 to 4 ± 2.5 mg Zn g⁻¹ DW and from 207.6 ± 24.5 to 45.4 ± 9.9 µg Cd g⁻¹ DW. However, at the end of the experiment bioaccumulations remained significantly higher than concentrations in control biofilms. Despite a diatom evolution in biofilm assemblages, taxonomic inventories did not demonstrate a complete restoration of diatom communities in biofilms under decontamination conditions compared with controls, since metal-resistant species initially present after colonisation at the contaminated site, such as Eolimna minima, persisted in high abundance in decontaminated biofilms. Biofilms kept under metal pressure showed very high bioaccumulation capacities and a sharp decline of species diversity which allowed identification of some resistant species. Regarding these first results on the behaviour of diatom biofilms under experimental decontamination conditions, improvement of the natural hydrosystem's chemical state appears quickly, but an eventual return to good ecological status appears delayed, with the persistence of metal-tolerant species even after 56 days.

Remediation of a watershed contaminated by heavy metals: a 2-year field biomonitoring of periphytic biofilms.

This study focuses on an industrial contamination site subjected to remediation processes since 2007 in the Riou-Mort watershed (southwest France). The purpose was to assess the first impacts of remediation on periphytic biofilms, and was performed during two years of biomonitoring. Periphytic biofilms were collected on glass slides immersed 24 days at different sites along the contamination gradient for 12 colonisation cycles. Metal contaminations (Cd and Zn) were analysed in biofilms and the evolution of diatom communities was assessed, integrating teratology quantifications. Despite remediation work initiated at the industrial site, this study demonstrated the persistence of metal contamination in water, as well as in biofilms. In addition, our data, showed that the remediation process was initially marked by an increase in metal contamination in the river, with increasing diatom community shifts. Metal-contaminated biofilms presented decreasing species diversities and were dominated by metal-resistant species such as Eolimna minima, whom abundances increased in 2010 reaching 57.2±10%. No significant decrease in metal accumulation was observed and total Cd content in biofilms collected downstream the industrial site ranged from 772.7±88 in July 2009 to 636.9±20 µg/gDW in July 2010. Results obtained on artificial substrates were compared with those of natural substrates and showed similar diatom communities and abundances of deformed diatoms but lower diversities. This ensured that glass slide subtrates gave a good representation of periphytic biofilm health. Finally, results were compared to studies performed before the remediation process and this did not reveal a decrease of metal accumulation in biofilms nor shifts in taxonomic composition of the communities, rather the remaining dominance of metal resistant species such as E. minima was confirmed.

Development of q-PCR approaches to assess water quality: effects of cadmium on gene expression of the diatom Eolimna minima.

This study was undertaken to develop molecular tools to assess water quality using diatoms as the biological model. Molecular approaches were designed following the development of a rapid and easy RNA extraction method suited to diatoms and the sequencing of genes involved in mitochondrial and photosystem metabolism. Secondly the impact of cadmium was evaluated at the genetic level by q-PCR on 9 genes of interest after exposure of Eolimna minima diatom populations cultured in suspension under controlled laboratory conditions. Their growth kinetics and Cd bioaccumulation were followed. Population growth rates revealed the high impact of Cd at 100 µg/L with total inhibition of growth. These results are linked to the high bioaccumulation values calculated after 14 days of exposure, 57.0±6.3 µg Cd/g dw and 734.1±70 µg Cd/g dw for exposures of 10 and 100 µg Cd/L respectively. Genetic responses revealed the impact of Cd on the mitochondrial metabolism and the chloroplast photosystem of E. minima exposed to 10 and 100 µg Cd/L with induction of cox1, 12S, d1 and psaA after 7 days of exposure for the concentration of 100 µg Cd/L and of nad5, d1 and psaA after 14 days of exposure for both conditions. This is the first reported use of q-PCR for the assessment of toxic pollution on benthic river diatoms. The results obtained presage interesting perspectives, but the techniques developed need to be optimized before the design of new water quality diagnosis tools for use on natural biofilms.

Comparison of periphytic biofilm and filter-feeding bivalve metal bioaccumulation (Cd and Zn) to monitor hydrosystem restoration after industrial remediation: a year of biomonitoring.

Despite a significant decrease in the metallic waste emissions from an industrial site and a remediation process initiated in 2007, the Riou-Mort watershed (southwest France) still exhibits high Cd and Zn concentrations. Metal wastes have long been proven to significantly disturb aquatic communities. In this study, bioaccumulation capacities and responses to the chemical improvement of the hydrosystem were assessed for a year along the contamination gradient through the comparison of two biological models: Corbicula fluminea and periphytic biofilms, both considered as good bioindicators. Bioaccumulation results confirmed the persistence of water contamination in Corbicula fluminea and biofilms with, respectively, maximum Cd concentrations reaching 80.6 and 861.2 µg gDW(-1), and Zn concentrations 2.0 and 21.3 mg gDW(-1). Biofilms exhibited bioaccumulation in close correlation with water contamination, while Corbicula fluminea presented Cd bioaccumulation clearly regulated by water temperature and metal concentrations, affecting the ventilatory activity, as revealed by condition indices measurements. Also, a linear regression using Cd bioaccumulation and temperature () showed that below approximately 6 °C Corbicula fluminea did not appear to accumulate metals. Bioconcentration factors (BCFs) were higher in biofilms in comparison with Corbicula fluminea and showed the great accumulation capacity of suspended particulate matter in biofilms. However, bioaccumulation capacities are known to be influenced by many factors other than metal concentrations, such as temperature, water oxygenation or plankton and nutrient concentrations. Thus, this study demonstrates the power of a combined assessment using both Corbicula fluminea and biofilms as bioindicators to give a more integrated view of water quality assessment. Finally, when comparing our results with previous studies, the start of hydrosystem restoration could be shown by decreasing bioaccumulation in organisms.