Exploring the subcellular distribution of mercury in green alga Chlamydomonas reinhardtii and diatom Cyclotella meneghiniana: A comparative study.

Mercury (Hg) is a priority pollutant of global concern because of its toxicity, its ability to bioaccumulate throughout the food web and reach significant concentrations in top predators. Phytoplankton bioconcentrate large amounts of Hg and play a key role in the entry of Hg into the aquatic food web. However, the subcellular distribution of Hg in freshwater phytoplankton, known to affect it toxicity and trophic transfer is understudied. The present study aimed at investigating the accumulation of inorganic Hg (iHg) and its subcellular distribution in freshwater phytoplankton species. To this end green alga Chlamydomonas reinhardtii and diatom Cyclotella meneghiniana were exposed to 10 and 100 nM of iHg for 2 h. The concentrations of Hg in the adsorbed, intracellular and subcellular (granules, debris, organelles, heat-stable peptides (HSP) and heat-denaturable proteins (HDP)) fractions were determined. The results showed that C. meneghiniana accumulated more Hg compared to C. reinhardtii at both iHg exposure concentrations (10 nM: 4.41 ± 0.74 vs. 1.10 ± 0.25 amol cell-1; 100 nM: 79.35 ± 10.78 vs. 38.31 ± 4.15 amol cell-1). The evaluation of the subcellular distribution of Hg, revealed that the majority of Hg was concentrated in the organelles fraction (59.7 % and 74.6 %) in the green algae. In the diatom, Hg was mainly found in the organelles (40.9 % and 33.3%) and in the HSP fractions (26.8 % and 40.1 %). The proportion of Hg in HDP fraction decreased in favor of the organelles fraction in C. reinhardtii when the exposure concentration increased, whereas the proportions in the debris and organelles fractions decreased in favor of HSP fraction in C. meneghiniana. This study provides pioneering information on the subcellular distribution of Hg within in freshwater phytoplankton, a knowledge that is essential to understand the toxicity and trophic transfer of Hg in contaminated aquatic environment.

Mercury species induce metabolic reprogramming in freshwater diatom Cyclotella meneghiniana.

Mercury is a hazardous pollutant of global concern. While advances have been made in identifying the detrimental effects caused by Hg species in phytoplankton, knowledge gaps remain regarding the metabolomic perturbations induced by inorganic mercury (Hg(II)) and monomethylmercury (MeHg) in these organisms. Diatoms represent a major phytoplankton group essential in various global biogeochemical cycles. The current study combined targeted metabolomics, bioaccumulation, and physiological response assays to investigate metabolic perturbations in diatom Cyclotella meneghiniana exposed for 2 h to nanomolar concentrations of Hg(II) and MeHg. Our findings highlight that such exposures induce reprogramming of the metabolism of amino acids, nucleotides, fatty acids, carboxylic acids and antioxidants. These alterations were primarily mercury-species dependent. MeHg exposure induced more pronounced reprogramming of the metabolism of diatoms than Hg(II), which led to less pronounced effects on ROS generation, membrane permeability and chlorophyll concentrations. Hg(II) treatments presented distinct physiological responses, with more robust metabolic perturbations at higher exposures. The present study provides first-time insights into the main metabolic alterations in diatom C. meneghiniana during short-term exposure to Hg species, deepening our understanding of the molecular basis of these perturbations.

Cytochrome c - silver nanoparticle interactions: Spectroscopy, thermodynamic and enzymatic activity studies.

Cytochrome c, an iron containing metalloprotein in the mitochondria of the cells with an oxide/redox property, plays key role in the cell apoptotic pathway. In this study, the interaction of silver nanoparticles (AgNPs) with cytochrome c (Cyt c) was investigated by using a combination of spectroscopic, imaging and thermodynamic techniques, including dynamic light scattering (DLS), ultraviolet-visible (UV-vis) spectroscopy, transmission electron microscopy (TEM), fluorescence spectroscopy, near and far circular dichroism (CD) spectroscopy, and isothermal titration calorimetry (ITC). DLS and UV-vis analysis evidenced the formation of surface complexes of Cyt c on AgNPs. The saturation of surface coverage of AgNPs was observed at 4.36 Cyt c molecules per nm2 of AgNPs. The surface complexation resulted in a promotion of the Ag dissolution overtime. The negative sign of enthalpic (ΔH) contribution suggested that electrostatic forces are indicative forces in the interaction between protein and AgNPs. Moreover, the fluorescence spectra revealed that the conformation of protein was altered around tryptophan (Trp) and tyrosine (Tyr) residues indicating the alteration of the tertiary structure of Cyt c. CD analysis evidenced that the secondary structure of Cyt c was modified under AgNPs-Cyt c interactions and the binding of Cyt c onto AgNPs resulted in remarkable structural perturbation around the active site heme, which in turn alter the protein enzymatic activity. The results of the present study contributed to a deeper insight on the mechanisms of interaction between NPs and biomacromolecules and could help establish the in vivo fate of AgNPs on cellular redox homeostasis.

Oxidative stress induced by sub-lethal exposure to copper as a mediator in development of bacterial resistance to antibiotics.

Limited information exists on how bacterial resistance to antibiotics is acquired and altered in response to short-term metal stress, and what the prevailing pathways are. Here the precursor mechanisms of development of bacterial antibiotic resistance mediated by oxidative stress induce under sub-lethal Cu2+ exposure were explored. The results showed that the overall level of antibiotic resistance in wild-type Escherichia coli and antibiotic-resistant E. coli was enhanced under 4 and 20 mg/L Cu2+ exposure, as demonstrated by the 2- to 8-fold increase in minimum inhibitory concentration (MIC). The MIC correlated with the increase of the cellular ROS generation and the enhancement of the antioxidant enzyme activity (p < 0.05), suggesting that changes in antibiotic resistance under sub-lethal Cu2+ exposure could be associated with oxidative stress. Likewise, enhanced cell membrane permeability and an increase in the number of bacteria entering the viable but non culturable (VBNC) state contributed to bacterial resistance to antibiotics. Moreover, the variance partitioning analysis demonstrated that the alterations of the antibiotic resistance phenotype of wild-type E. coli was mainly caused by oxidative stress-mediated increase in cell membrane permeability and entry into the VBNC state. The development of antibiotic resistance in resistant E. coli was primarily attributed to changes in the abundance and horizontal transfer ability of its antibiotic resistance genes, both of which contributed up to 20 %. Taken together the results allowed to propose a conseptual scheme on developing bacterial antibiotic resistance mediated by oxidative stress under sub-lethal Cu2+ exposure. This result provided a strong basis for reduction of early bacterial resistance.

Aquatic organisms modulate the bioreactivity of engineered nanoparticles: focus on biomolecular corona.

The increased use of nanoparticle (NP)-enabled materials in everyday-life products have raised concerns about their environmental implications and safety. This motivated the extensive research in nanoecotoxicology showing the possibility that NPs could cause harm to the aquatic organisms if present at high concentrations. By contrast, studies dealing with influence that organisms could exert on the fate and thus effects of NPs are still very rare. Drawing on the existing up-to-date knowledge we critically discuss the formation of biomolecular corona as one of the mechanisms by which organisms exerted control on the NPs fate in the aquatic and biotic environments. We focused the formation of corona by exogeneous and endogenous biomolecules and illustrated the discussion with the specific example of phytoplankton and aquatic invertebrate species. We highlighted the necessity to incorporate the concept of biomolecular corona within more general framework considering the feedback of aquatic organisms and the control they exert in shaping the fate and impact of NPs in the aquatic and biological environment. In our view such broader perspective will contribute to get novel insights into the drivers of environmental transformations of NPs and their mechanisms, which are important in environmental risk assessment.

Metabolic alterations in alga Chlamydomonas reinhardtii exposed to nTiO2 materials.

Nano-sized titanium dioxide (nTiO2) is one of the most commonly used materials, however the knowledge about the molecular basis for metabolic and physiological changes in phytoplankton is yet to be explored. In the present study we use a combination of targeted metabolomics, transcriptomics and physiological response studies to decipher the metabolic perturbation in green alga Chlamydomonas reinhardtii exposed for 72 h to increasing concentrations (2, 20, 100 and 200 mg L-1) of nTiO2 with primary sizes of 5, 15 and 20 nm. Results show that the exposure to all three nTiO2 materials induced perturbation of the metabolism of amino acids, nucleotides, fatty acids, tricarboxylic acids, antioxidants but not in the photosynthesis. The alterations of the most responsive metabolites were concentration and primary size-dependent despite the significant formation of micrometer-size aggregates and their sedimentation. The metabolic perturbations corroborate the observed physiological responses and transcriptomic results and confirmed the importance of oxidative stress as a major toxicity mechanism for nTiO2. Transcriptomics revealed also an important influence of nTiO2 treatments on the transport, adenosine triphosphate binding cassette transporters, and metal transporters, suggesting a perturbation in a global nutrition of the microalgal cell, which was most pronounced for exposure to 5 nm nTiO2. The present study provides for the first-time evidence for the main metabolic perturbations in green alga C. reinhardtii exposed to nTiO2 and helps to improve biological understanding of the molecular basis of these perturbations.

Comparative study of the sensitivity of two freshwater gastropods, Lymnaea stagnalis and Planorbarius corneus, to silver nanoparticles: bioaccumulation and toxicity.

Metal-based nanoparticles (NPs) are considered detrimental to aquatic organisms due to their potential accumulation. However, little is known about the mechanisms underlying these effects and their species-specificity. Here we used stable silver (Ag) NPs (20 nm, from 10 to 500 µg/L) with a low dissolution rate (≤2.4%) to study the bioaccumulation and biological impacts in two freshwater gastropods: Lymnaea stagnalis and Planorbarius corneus. No mortality was detected during the experiments. Ag bioaccumulation showed a dose-related increase with an enhanced concentration in both species after 7d exposure. L. stagnalis displayed a higher accumulation for AgNPs than P. corneus (e.g., up to 18- and 15-fold in hepatopancreas and hemolymph, respectively) which could be due to the more active L. stagnalis having greater contact with suspended AgNPs. Furthermore, the hepatopancreas and stomach were preferred organs for bioaccumulation compared to the kidney, mantle and foot. Regarding biological responses, the hemolymph rather than hepatopancreas appeared more susceptible to oxidative stress elicited by AgNPs, as shown by significantly increasing lipid peroxidation (i.e., formation of malondialdehyde). Neurotoxicity was detected in L. stagnalis when exposed to high concentrations (500 µg/L). Comparison with impacts elicited by dissolved Ag revealed that the effects observed on AgNPs exposure were mainly attributable to NPs. These results highlighted the relationship between the physiological traits, bioaccumulation, and toxicity responses of these two species to AgNPs and demonstrated the necessity of species-specificity considerations when assessing the toxicity of NPs.

Simple Acid Digestion Procedure for the Determination of Total Mercury in Plankton by Cold Vapor Atomic Fluorescence Spectroscopy.

Plankton, at the bottom of the food web, play a central role in the entry of mercury into the aquatic biota. To investigate their role in mercury uptake, reliable analytical procedures for Hg analysis are highly sought. Wet digestion procedures for determining total mercury in different biological matrices have been established since years, however only few studies focused on planktonic samples. In the present work, a simple and cost-effective wet digestion method was developed for the determination of total mercury in samples of small plankton material using a cold vapor atomic fluorescence spectroscopy (CVAFS). The optimization of the digestion method was achieved by using glass vessels with Teflon caps, low amount of acids (3 mL w/w 65% HNO3 or 3 mL 50% v/v HNO3), a constant temperature of 85 °C, the presence and absence of pre-ultrasound treatment, and a continuous digestion period (12 h). Certified reference materials IAEA-450 (unicellular alga Scenedesmus obliquus) and BRC-414 (plankton matrix) were used to optimize and validate the digestion method. The recovery efficiency of the proposed method for IAEA-450 and BCR-414 (3.1 mg and 21.5 mg) ranged between 94.1 ± 7.6% and 97.2 ± 4.6%. The method displayed a good recovery efficiency and precision for plankton matrices of low size. Thus, allowing better digestion of planktonic samples for mercury analysis using CVAFS techniques.

Asymmetrical Flow Field-Flow Fractionation Methods for Quantitative Determination and Size Characterization of Thiols and for Mercury Size Speciation Analysis in Organic Matter-Rich Natural Waters.

Asymmetrical flow field-flow fractionation (AF4) efficiently separates various macromolecules and nano-components of natural waters according to their hydrodynamic sizes. The online coupling of AF4 with fluorescence (Fluo) and UV absorbance (UV) detectors (FluoD and UVD, respectively) and inductively coupled plasma-mass spectrometry (ICP-MS) provides multidimensional information. This makes it a powerful tool to characterize and quantify the size distributions of organic and inorganic nano-sized components and their interaction with trace metals. In this study, we developed a method combining thiol labeling by monobromo(trimethylammonio)bimane bromide (qBBr) with AF4-FluoD to determine the size distribution and the quantities of thiols in the macromolecular dissolved organic matter (DOM) present in highly colored DOM-rich water sampled from Shuya River and Lake Onego, Russia. We found that the qBBr-labeled components of DOM (qB-DOM) were of humic type, characterized by a low hydrodynamic size (d h < 2 nm), and have concentrations <0.3 µM. After enrichment with mercury, the complexes formed between the nano-sized components and Hg were analyzed using AF4-ICP-MS. The elution profile of Hg followed the distribution of the UV-absorbing components of DOM, characterized by slightly higher sizes than qB-DOM. Only a small proportion of Hg was associated with the larger-sized components containing Fe and Mn, probably inorganic oxides that were identified in most of the samples from river to lake. The size distribution of the Hg-DOM complexes was enlarged when the concentration of added Hg increased (from 10 to 100 nM). This was explained by the presence of small iron oxides, overlapping the size distribution of Hg-DOM, on which Hg bound to a small proportion. In addition, to provide information on the dispersion of macromolecular thiols in colored DOM-rich natural water, our study also illustrated the potential of AF4-FluoD-UVD-ICP-MS to trace or quantify dynamic changes while Hg binds to the natural nano-colloidal components of surface water.

Asymmetrical Flow Field-Flow Fractionation Coupled to ICP-MS for Characterization of Trace Metal Species in the Environment from Macromolecular to Nano-Assemblage Forms: Current Challenges for Quantification.

Asymmetrical flow field-flow fractionation (AF4) is a powerful technique employed for the separation of macromolecules, nanoparticles, and their assemblages according to their hydrodynamic behavior. It is well known that at this size range, complex interactions can occur between components (e.g. surface adsorption, aggregation) controlling the fate of trace metals (TMs) bound to them. AF4 coupling to inductively coupled plasma mass spectrometry (ICP-MS) allows the quantification of metal-containing species at trace levels present in environmental and biological systems on a size-composition basis. The combination of AF4-ICP-MS with other online detectors provides additional information that allows the assessment of the origin of analytes present in mixtures and complex matrixes with minimal sample preparation, which is crucial for understanding the behavior of trace metal contaminants. Despite the increasing use of AF4-ICP-MS in environmental contexts, we acknowledge that the quantification of inorganic species using such combined techniques requires further development of standardized procedures and need certified reference materials. In this review, we also discuss critical endpoints within the ICP-MS instrument coupled to AF4 that need to be controlled before quantitative measurements can be validated. Then, we illustrate how the combination of different online detectors in addition to ICP-MS offers an integrated picture of natural components states, thus providing key information on the changes in behavior of trace metal species and metallic nanoparticles (MNPs) as observed in both environmental samples and biofluids.

Light-trapped caddisflies to decipher the role of species traits and habitats in Hg accumulation and transfer.

We present a novel meta-community approach to explore the influence of species traits, such as adult body size, larval feeding type and microhabitat, as well as larval macrohabitat (main river channel vs. floodplain water bodies) on the concentration of total Hg accumulated ([THg]) in assemblages of adult caddisflies. We analyzed [THg] in 157 light-trapped adult caddisflies in a floodplain sector of the French upper Rhône River and used a linear mixed effect model to decipher the role of species traits and habitats in Hg accumulation. Variation of [THg] between species was best explained by the larval feeding type, whereas the contributions of adult size and larval micro and macro-habitat were minor. Results showed that [THg] in species associated with floodplain macrohabitats in the larval stage was lower than in those associated with the main river channel. This difference could depend on complexation of Hg by DOM (in the floodplain) and MES (in the main channel). This research provides a first evidence of the potential of an entire caddisfly assemblage for the assessment of contamination in large alluvial rivers. The implications of the results are discussed in view of the possible role of caddisflies as vectors of Hg to riparian predators.

Dual role of titanium dioxide nanoparticles in the accumulation of inorganic and methyl mercury by crustacean Daphnia magna through waterborne and dietary exposure.

Titanium dioxide nanoparticles (nTiO2) are widely used in numerous products, yet their role in the accumulation and transfer of other contaminants in the aquatic food webs is not well understood. The influence of nTiO2 on inorganic (IHg) and monomethyl mercury (MeHg) accumulation in invertebrate Daphnia magna through waterborne and dietary exposure was thus thoroughly investigated. The results showed that nTiO2 led to a substantial decrease of the total mercury body burden (THg) in D. magna in direct waterborne exposure to IHg/MeHg. However, exposure to nTiO2 pre-treated with IHg/MeHg resulted in an increase of the THg body burden in daphnids. The presence of nTiO2 led to a substantial decrease of the THg in D. magna when exposed to IHg/MeHg via algal food. These effects were more pronounced for IHg than that for MeHg due to the higher adsorption capabilities of nTiO2 for IHg. In addition, high concentrations of nTiO2 favored the trophic transfer of IHg/MeHg through feeding on nTiO2 pre-treated with Hg, however lessened it when D. magna were fed on alga pre-treated with IHg/MeHg. Comparable assimilation efficiency (AE), determined as Hg retained in daphnids after depuration, was observed in D. magna when exposed to IHg/MeHg via algal food regardless the absence or presence of 20 mgL-1 nTiO2. By contrast, an increase of the AE of MeHg through feeding on nTiO2 and alga was found in the presence of higher concentration of 200 mgL-1 nTiO2. The present results will help to better understand the role of nTiO2 on bioavailability and trophic transfer of global contaminants, such as mercury, known to bioaccumulate and biomagnify in the aquatic environment.

Determination of the Intracellular Complexation of Inorganic and Methylmercury in Cyanobacterium Synechocystis sp. PCC 6803.

Understanding of mercury (Hg) complexation with low molecular weight (LMW) bioligands will help elucidate its speciation. In natural waters, the rate of this complexation is governed by physicochemical, geochemical, and biochemical parameters. However, the role of bioligands involved in Hg intracellular handling by aquatic microorganisms is not well documented. Here, we combine the use of isotopically labeled Hg species (inorganic and monomethylmercury, iHg and MeHg) with gas or liquid chromatography coupling to elemental and molecular mass spectrometry to explore the role of intracellular biogenic ligands involved in iHg and MeHg speciation in cyanobacterium Synechocystis sp. PCC 6803, a representative phytoplankton species. This approach allowed to track resulting metabolic and newly found intracellular Hg biocomplexes (e.g., organic thiols) in Synechocystis sp. PCC 6803 finding different intracellular Hg species binding affinities with both high and low molecular weight (HMW and LMW) bioligands in the exponential and stationary phase. Furthermore, the parallel detection with both elemental and molecular ionization sources allowed the sensitive detection and molecular identification of glutathione (GSH) as the main low molecular weight binding ligand to iHg ((GS)2-Hg) and MeHg (GS-MeHg) in the cytosolic fraction. Such a novel experimental approach expands our knowledge on the role of biogenic ligands involved in iHg and MeHg intracellular handling in cyanobacteria.

Species-specific isotope tracking of mercury uptake and transformations by pico-nanoplankton in an eutrophic lake.

The present study aims to explore the bioaccumulation and biotic transformations of inorganic (iHg) and monomethyl mercury (MMHg) by natural pico-nanoplankton community from eutrophic lake Soppen, Switzerland. Pico-nanoplankton encompass mainly bacterioplankton, mycoplankton and phytoplankton groups with size between 0.2 and 20 µm. Species-specific enriched isotope mixture of 199iHg and 201MMHg was used to explore the accumulation, the subcellular distribution and transformations occurring in natural pico-nanoplankton sampled at 2 different depths (6.6 m and 8.3 m). Cyanobacteria, diatoms, cryptophyta, green algae and heterotrophic microorganisms were identified as the major groups of pico-nanoplankton with diatoms prevailing at deeper samples. Results showed that pico-nanoplankton accumulated both iHg and MMHg preferentially in the cell membrane/organelles, despite observed losses. The ratios between the iHg and MMHg concentrations measured in the membrane/organelles and cytosol were comparable for iHg and MMHg. Pico-nanoplankton demethylate added 201MMHg (~4 and 12% per day depending on cellular compartment), although the involved pathways are to further explore. Comparison of the concentrations of 201iHg formed from 201MMHg demethylation in whole system, medium and whole cells showed that 82% of the demethylation was biologically mediated by pico-nanoplankton. No significant methylation of iHg by pico-nanoplankton was observed. The accumulation of iHg and MMHg and the percentage of demethylated MMHg correlated positively with the relative abundance of diatoms and heterotrophic microorganisms in the pico-nanoplankton, the concentrations of TN, Mg2+, NO3-, NO2-, NH4+ and negatively with the concentrations of DOC, K+, Na+, Ca2+, SO42-. Taken together the results of the present field study confirm the role of pico-nanoplankton in Hg bioaccumulation and demethylation, however further research is needed to better understand the underlying mechanisms and interconnection between heterotrophic and autotrophic microorganisms.

Metabolomic Responses of Green Alga Chlamydomonas reinhardtii Exposed to Sublethal Concentrations of Inorganic and Methylmercury.

Metabolomics characterizes low-molecular-weight molecules involved in different biochemical reactions and provides an integrated assessment of the physiological state of an organism. By using liquid chromatography-mass spectrometry targeted metabolomics, we examined the response of green alga Chlamydomonas reinhardtii to sublethal concentrations of inorganic mercury (IHg) and monomethylmercury (MeHg). We quantified the changes in the levels of 93 metabolites preselected based on the disturbed metabolic pathways obtained in a previous transcriptomics study. Metabolites are downstream products of the gene transcription; hence, metabolite quantification provided information about the biochemical status of the algal cells exposed to Hg compounds. The results showed that the alga adjusts its metabolism during 2 h exposure to 5 × 10-9 and 5 × 10-8 mol L-1 IHg and MeHg by increasing the level of various metabolites involved in amino acid and nucleotide metabolism, photorespiration, and tricarboxylic acid (TCA) cycle, as well as the metabolism of fatty acids, carbohydrates, and antioxidants. Most of the metabolic perturbations in the alga were common for IHg and MeHg treatments. However, the exposure to IHg resulted in more pronounced perturbations in the fatty acid and TCA metabolism as compared with the exposure to MeHg. The observed metabolic perturbations were generally consistent with our previously published transcriptomics results for C. reinhardtii exposed to the comparable level of IHg and MeHg. The results highlight the potential of metabolomics for toxicity evaluation, especially to detect effects at an early stage of exposure prior to their physiological appearance.

Distinguishing the effects of Ce nanoparticles from their dissolution products: identification of transcriptomic biomarkers that are specific for ionic Ce in Chlamydomonas reinhardtii.

Cerium (Ce) is a rare earth element that is incorporated in numerous consumer products, either in its cationic form or as engineered nanoparticles (ENPs). Given the propensity of small oxide particles to dissolve, it is unclear whether biological responses induced by ENPs will be due to the nanoparticles themselves or rather due to their dissolution. This study provides the foundation for the development of transcriptomic biomarkers that are specific for ionic Ce in the freshwater alga, Chlamydomonas reinhardtii, exposed either to ionic Ce or to two different types of small Ce ENPs (uncoated, ∼10 nm, or citrate-coated, ∼4 nm). Quantitative reverse transcription PCR was used to analyse mRNA levels of four ionic Ce-specific genes (Cre17g.737300, MMP6, GTR12, and HSP22E) that were previously identified by whole transcriptome analysis in addition to two oxidative stress biomarkers (APX1 and GPX5). Expression was characterized for exposures to 0.03-3 µM Ce, for 60-360 min and for pH 5.0-8.0. Near-linear concentration-response curves were obtained for the ionic Ce and as a function of exposure time. Some variability in the transcriptomic response was observed as a function of pH, which was attributed to the formation of metastable Ce species in solution. Oxidative stress biomarkers analysed at transcriptomic and cellular levels confirmed that different effects were induced for dissolved Ce in comparison to Ce ENPs. The measured expression levels confirmed that changes in Ce speciation and the dissolution of Ce ENPs greatly influence Ce bioavailability.

Microbial community diversity and composition in river sediments contaminated with tetrabromobisphenol A and copper.

The microbial community composition in aquatic ecosystems have received increased attention. However, the knowledge gap relative to the responses of bacterial, archaeal and fungal communities in co-contaminated river sediments remain poorly studied. Here, we investigated the changes of tetrabromobisphenol A (TBBPA) and copper (Cu) concentrations and the responses of microbial communities in co-contaminated sediments during long-term incubation. TBBPA concentrations significantly decreased over time, whereas Cu concentrations remained relatively stable over the 60-day incubation. Abundances of the bacterial 16S rRNA, archaeal 16S rRNA and fungal ITS genes ranged from 6.53 × 106 to 1.26 × 109 copies g-1, 1.12 × 106 to 5.47 × 106 copies g-1 and 5.33 × 103 to 7.51 × 104 copies g-1 in the samples, respectively. A total of 11, 6 and 5 bacterial, archaeal and fungal phyla were identified across all samples. Bacterial, archaeal and fungal communities mainly consisted of members from the phyla Proteobacteria and Acidobacteria, Methanomicrobia and Woesearchaeia as well as Agaricales and Helotiales, respectively. Fungal communities showed a stronger response to pollutant addition after a long incubation compared with bacterial and archaeal communities. The variance analysis results revealed that the bacterial, archaeal and fungal microbial communities of all treatments were distinctly distributed into two separated clusters according to incubation time. However, the three microbial communities did not significantly change in response to pollutant types, which was consistent with variation in relative abundances of the three microbial communities. These findings improve our understanding of the ecotoxicological effects of co-exposure on sediment microbial communities.

Morphological plasticity in Chlamydomonas reinhardtii and acclimation to micropollutant stress.

Phytoplankton are characterized by a great phenotypic plasticity and amazing morphological variability, both playing a primary role in the acclimation to changing environments. However, there is a knowledge gap concerning the role of algal morphological plasticity in stress responses and acclimation to micropollutants. The present study aims at examining palmelloid colony formation of the green alga Chlamydomonas reinhardtii upon micropollutants exposure. Cells were exposed to four micropollutants (MPs, copper, cadmium, PFOS and paraquat) with different modes of action for a duration of 72 h. Effects of MPs on palmelloid formation, growth and physiological traits (chlorophyll fluorescence, membrane integrity and oxidative stress) were monitored by flow cytometry and fluorescence microscopy. Palmelloid formation was observed upon treatment with the four micropollutants. Number of palmelloid colonies and their size were dependent on MP concentration and exposure duration. Cells reverted to their unicellular lifestyle when colonies were harvested and inoculated in fresh medium indicating that palmelloid formation is a plastic response to micropollutants. No physiological effects of these compounds were observed in cells forming palmelloids. Palmelloid colonies accumulated lower Cd concentration than unicellular C. reinhardtii suggesting that colony formation protects the cells from MPs stress. The results show that colony formation in Chlamydomonas reinhardtii is a stress response strategy activated to face sub-lethal micropollutant concentrations.

Academic expertise in assisting private companies in the fields of environment and environmental toxicology: the role of individual expertise.

The scientific knowledge produced by academic research can be valued in all sectors of human activity, including private sector. The ROVALTAIN Foundation organized a round-table during its scientific day in 2019. It crossed the points of view of academic scientists and industrial partners, addressing five main topics. The first one concerned the validation of a common definition of the academic research/private partners interface. Then, the group discussed the place for academic expertise in the corporate world; the advantages of involving academic researchers in expertise for the private sector; and the limits of this model. To conclude, the need of a third party, like the ROVALTAIN Foundation, as a catalyzer in building the interface between academic research and private partners has been discussed.

The interplay of flow processes shapes aquatic invertebrate successions in floodplain channels - A modelling applied to restoration scenarios.

The high biotic diversity supported by floodplains is ruled by the interplay of geomorphic and hydrological processes at various time scales, from daily fluctuations to decennial successions. Because understanding such processes is a key question in river restoration, we attempted to model changes in taxonomic richness in an assemblage of 58 macroinvertebrate taxa (21 gastropoda and 37 ephemeroptera, plecoptera and trichoptera, EPT) along two successional sequences typical for former braided channels. Individual models relating the occurrence of taxa to overflow and backflow durations were developed from field measurements in 19 floodplain channels of the Rhône floodplain (France) monitored over 10 years. The models were combined to simulate diversity changes along a progressive alluviation and disconnection sequence after the reconnection with the main river of a previously isolated channel. Two scenarios were considered: (i) an upstream + downstream reconnection creating a lotic channel, (ii) a downstream reconnection creating a semi-lotic channel. Reconnection led to a direct increase in invertebrate richness (on average x2.5). However, taxonomical richness showed a constant decrease as isolation progressed and reached an average of 2 for EPT and 7 for gastropods at the end of the scenarios. With more than 80% of the taxonomic models with an AUC equal or higher than 0.7 and slopes of linear relations between observed and predicted richness of 0.75 (gastropods) and 1 (EPT), the Boosted Regression Trees (BRT) provided a good basis for prediction of species assemblages. These models can be used to quantify a priori the sustainability and ecological efficiency of restoration actions and help floodplain restoration planning and management.