DRomics: A Turnkey Tool to Support the Use of the Dose-Response Framework for Omics Data in Ecological Risk Assessment.

Omics approaches (e.g., transcriptomics, metabolomics) are promising for ecological risk assessment (ERA) since they provide mechanistic information and early warning signals. A crucial step in the analysis of omics data is the modeling of concentration-dependency which may have different trends including monotonic (e.g., linear, exponential) or biphasic (e.g., U shape, bell shape) forms. The diversity of responses raises challenges concerning detection and modeling of significant responses and effect concentration (EC) derivation. Furthermore, handling high-throughput data sets is time-consuming and requires effective and automated processing routines. Thus, we developed an open source tool (DRomics, available as an R-package and as a web-based service) which, after elimination of molecular responses (e.g., gene expressions from microarrays) with no concentration-dependency and/or high variability, identifies the best model for concentration-response curve description. Subsequently, an EC (e.g., a benchmark dose) is estimated from each curve, and curves are classified based on their model parameters. This tool is especially dedicated to manage data obtained from an experimental design favoring a great number of tested doses rather than a great number of replicates and also to handle properly monotonic and biphasic trends. The tool finally provides restitution for a table of results that can be directly used to perform ERA approaches.

Hierarchical modelling of species sensitivity distribution: development and application to the case of diatoms exposed to several herbicides.

The species sensitivity distribution (SSD) is a key tool to assess the ecotoxicological threat of contaminants to biodiversity. For a contaminant, it predicts which concentration is safe for a community of species. Widely used, this approach suffers from several drawbacks: (i) summarizing the sensitivity of each species by a single value entails a loss of valuable information about the other parameters characterizing the concentration-effect curves; (ii) it does not propagate the uncertainty on estimated sensitivities into the SSD; (iii) the hazardous concentration estimated with SSD only indicates the threat to biodiversity, without any insight about a global response of the community related to the measured endpoint. To remedy these drawbacks, we built a global hierarchical model including the concentration-effect model together with the distribution law of the SSD. We revisited the current SSD approach to account for more sources of variability and uncertainty into the prediction than the traditional analysis and to assess a global response for the community. Working within a Bayesian framework, we were able to compute an SSD taking into account the uncertainty from the original raw data. We also developed a quantitative indicator of a global response of the community to the contaminant. We applied this methodology to study the toxicity and the risk of six herbicides to benthic diatoms from Lake Geneva, based on the biomass endpoint. Our approach highlighted a wide variability within the set of diatom species for all the parameters of the concentration-effect model and a potential correlation between them. Remarkably, variability of the shape parameter of the model and correlation had not been considered before. Comparison between the SSD and the global response of the community revealed that protecting 95% of the species might preserve only 80-86% of the global response. Finally, propagating the uncertainty on the estimated sensitivity showed that building an SSD on a low level of effect, such as EC10, might be unreasonable as it induces a large uncertainty on the result.

Linking diatom sensitivity to herbicides to phylogeny: a step forward for biomonitoring?

Phylogeny has not yet been fully accepted in the field of ecotoxicology, despite studies demonstrating its potential for developing environmental biomonitoring tools, as it can provide an a priori assessment of the sensitivity of several indicator organisms. We therefore investigated the relationship between phylogeny and sensitivity to herbicides in freshwater diatom species. This study was performed on four photosystem II inhibitor herbicides (atrazine, terbutryn, diuron, and isoproturon) and 14 diatom species representative of Lake Geneva biofilm diversity. Using recent statistical tools provided by phylogenetics, we observed a strong phylogenetic signal for diatom sensitivity to herbicides. There was a major division in sensitivity to herbicides within the phylogenetic tree. The most sensitive species were mainly centrics and araphid diatoms (in this study, Thalassiosirales and Fragilariales), whereas the most resistant species were mainly pennates (in this study, Cymbellales, Naviculales, and Bacillariales). However, there was considerable variability in diatom sensitivity within the raphid clade, which could be explained by differences in trophic preferences (autotrophy or heterotrophy). These traits appeared to be complementary in explaining the differences in sensitivity observed at a refined phylogenetic level. Using phylogeny together with complementary traits, as trophic preferences, may help to predict the sensitivity of communities with a view to protecting their ecosystem.

Seasonal shift in the sensitivity of a natural benthic microalgal community to a herbicide mixture: impact on the protective level of thresholds derived from species sensitivity distributions.

Seasonal changes in the structure and composition of a benthic microalgal community may lead to different responses to herbicide contamination during different seasons. Consequently, the thresholds derived from risk assessment tools such as species sensitivity distributions (SSDs) must allow for these changes. We built a single-substance SSD for each of four herbicides (atrazine, terbutryn, diuron and isoproturon), which was specific to the sensitivity of the benthic diatoms found in Lake Geneva, in order to derive protective thresholds for a mixture of these four herbicides using the concentration addition model. We then investigated (1) the structural parameters of a Lake Geneva benthic microalgal community during two contrasting seasons (summer 2012 and winter 2013), (2) the response of these communities to a herbicide mixture, and (3) the protective levels of the thresholds derived. The winter community was characterized by having greater biomass, diatom species richness, and diversity metrics, and lower non-diatom species richness than the summer community. The differences in the diatom communities composition in these seasons appeared to be primarily driven by the environmental nitrate concentrations and the temperature. Moreover, the species in the winter community were more resistant to herbicides than those found in the summer community. Consequently, the protective threshold for this herbicide mixture obtained in this study was in fact protective for the winter community, but not for the summer community based on their structural parameters. Thus, the protective level against herbicides of the threshold for the benthic microalgal community should take into account changes in the environmental physico-chemical conditions that strongly influence the structure and composition of the community. The fact that the succession of species over time (i.e., over the seasons) is difficult to predict introduces uncertainties into the estimation of protective thresholds and questions their applicability year round.

The effect of temperature and a herbicide mixture on freshwater periphytic algae.

Temperature is a strong driver of biofilm formation and of the dynamics of microalgae in freshwater. Moreover, exposure to herbicides is a well-known stressor of periphytic communities in anthropized aquatic environments. We tested these two environmental factors on periphytic communities that had been sampled from the littoral zone of Lake Geneva and acclimatized in the lab for 3 weeks at 18, 21, 24 and 28 °C. After this acclimation period, differences in the composition of the diatom community and decreases in cell density were observed corresponding to the temperature gradient. These acclimated communities were then exposed to 23 and 140 nM of a mixture composed of equitoxic quantities of atrazine, terbutryn, diuron and isoproturon. The periphytic community was more sensitive to the herbicide mixture at 18 °C than at higher temperatures, suggesting that higher temperature reduced its toxicity. Small and pioneer diatom species known to be promoted by contamination also appeared to benefit from higher temperatures. Temperature therefore appears to condition the herbicide sensitivity of periphytic communities.

Linking ecotoxicological effects on biodiversity and ecosystem functions to impairment of ecosystem services is a challenge: an illustration with the case of plant protection products.

There is growing interest in using the ecosystem services framework for environmental risk assessments of chemicals, including plant protection products (PPPs). Although this topic is increasingly discussed in the recent scientific literature, there is still a substantial gap between most ecotoxicological studies and a solid evaluation of potential ecotoxicological consequences on ecosystem services. This was recently highlighted by a collective scientific assessment (CSA) performed by 46 scientific experts who analyzed the international science on the impacts of PPPs on biodiversity, ecosystem functions, and ecosystem services. Here, we first point out the main obstacles to better linking knowledge on the ecotoxicological effects of PPPs on biodiversity and ecological processes with ecosystem functions and services. Then, we go on to propose and discuss possible pathways for related improvements. We describe the main processes governing the relationships between biodiversity, ecological processes, and ecosystem functions in response to effects of PPP, and we define categories of ecosystem functions that could be directly linked with the ecological processes used as functional endpoints in investigations on the ecotoxicology of PPPs. We then explore perceptions on the possible links between these categories of ecosystem functions and ecosystem services among a sub-panel of the scientific experts from various fields of environmental science. We find that these direct and indirect linkages still need clarification. This paper, which reflects the difficulties faced by the multidisciplinary group of researchers involved in the CSA, suggests that the current gap between most ecotoxicological studies and a solid potential evaluation of ecotoxicological consequences on ecosystem services could be partially addressed if concepts and definitions related to ecological processes, ecosystem functions, and ecosystem services were more widely accepted and shared within the ecotoxicology community. Narrowing this gap would help harmonize and extend the science that informs decision-making and policy-making, and ultimately help to better address the trade-off between social benefits and environmental losses caused by the use of PPPs.

Community metabolomics provides insights into mechanisms of pollution-induced community tolerance of periphyton.

Chemical pollution is a major concern for freshwater ecosystems, but the impact and mechanisms of chemical stressors on communities are barely understood. Pollution stress beyond natural homeostatic capacities can trigger succession of tolerant species within a community, enhancing the overall community tolerance. This process was operationalized in the Pollution-Induced Community Tolerance (PICT) concept and applied in many case studies, however, the molecular mechanisms of community tolerance and implications for ecological functions remain largely unexplored. Our study aimed to demonstrate that 1) community metabolomics can unravel potential mechanisms of PICT in periphyton and 2) induced tolerance helps to maintain primary production under re-occuring pollution. To this end, we grew periphyton for 5 weeks with and without the model herbicide diuron in microcosms, quantified PICT, and determined the related metabolic fingerprint of periphyton by GC-MS-based untargeted metabolomics. Further, we explored the autotrophic community based on pigment composition and functional parameters including photosynthesis and gross primary production. Chronic diuron exposure resulted in a shift in pigment composition, higher community tolerance and an individual metabolic fingerprint in the contaminated communities. Opposing responses of selected metabolites during a short-term exposure indicated differences in diuron pre-adaptation in the different communities. Metabolites (threonic acid and two sugar acid lactones) were found to be related to tolerance development, suggesting that ascorbate metabolism was induced in contaminated communities. Despite these compensating mechanism, contaminated communities were compromised in production-to-respiration ratio and biomass. A ranking of sensitivity thresholds of different biological endpoints revealed that metabolites were less sensitive than photosynthetic parameters, which reflects the mode-of-action of the herbicide. In conclusion, we could demonstrate that community metabolomics is able to unravel complex biochemical changes and allows mechanistic insights into community tolerance. Moreover, we were able to show that induced community tolerance was insufficient to safeguard functions like primary production.

A critical review of effect modeling for ecological risk assessment of plant protection products.

A wide diversity of plant protection products (PPP) is used for crop protection leading to the contamination of soil, water, and air, which can have ecotoxicological impacts on living organisms. It is inconceivable to study the effects of each compound on each species from each compartment, experimental studies being time consuming and cost prohibitive, and animal testing having to be avoided. Therefore, numerous models are developed to assess PPP ecotoxicological effects. Our objective was to provide an overview of the modeling approaches enabling the assessment of PPP effects (including biopesticides) on the biota. Six categories of models were inventoried: (Q)SAR, DR and TKTD, population, multi-species, landscape, and mixture models. They were developed for various species (terrestrial and aquatic vertebrates and invertebrates, primary producers, micro-organisms) belonging to diverse environmental compartments, to address different goals (e.g., species sensitivity or PPP bioaccumulation assessment, ecosystem services protection). Among them, mechanistic models are increasingly recognized by EFSA for PPP regulatory risk assessment but, to date, remain not considered in notified guidance documents. The strengths and limits of the reviewed models are discussed together with improvement avenues (multigenerational effects, multiple biotic and abiotic stressors). This review also underlines a lack of model testing by means of field data and of sensitivity and uncertainty analyses. Accurate and robust modeling of PPP effects and other stressors on living organisms, from their application in the field to their functional consequences on the ecosystems at different scales of time and space, would help going toward a more sustainable management of the environment. Graphical Abstract Combination of the keyword lists composing the first bibliographic query. Columns were joined together with the logical operator AND. All keyword lists are available in Supplementary Information at https://doi.org/10.5281/zenodo.5775038 (Larras et al. 2021).

Federating young researchers in microbial ecotoxicology: EcotoxicoMicYR 2021, the first international webinar organized for and by young microbial ecotoxicology researchers.

The EcotoxicoMicYR group was initially composed of 4 Ph.D. students and 4 post-doctoral researchers. In brief, the EcotoxicoMicYR webinar took place three Monday afternoons in a row from November 22 to December 6, 2021. These three half-day webinars reached a success beyond our expectations with 25 countries and 41 presentations. Keynote lectures were delivered by Dr Fabio Roldan (Pontificia Universidad Javeriana, Colombia), Dr Belinda Ferrari (The University of New South Wales, Australia), and Dr Ahmed Tlili (Eawag, Switzerland). Their presentations provided an insight on latest research developments in the microbial ecotoxicology field and highlighted their specific contribution to this discipline. Twenty-two oral presentations and 16 pre-recorded presentations were diffused.

A meta-analysis of ecotoxicological models used for plant protection product risk assessment before their placing on the market.

Before their placing on the market, the safety of plant protection products (PPP) towards both human and animal health, and the environment has to be assessed using experimental and modelling approaches. Models are crucial tools for PPP risk assessment and some even help to avoid animal testing. This review investigated the use of modelling approaches in the ecotoxicology section of PPP active substance assessment reports prepared by the authorities and opened to consultation from 2011 to 2021 in the European Union. Seven categories of models (Structure-Activity, ToxicoKinetic, ToxicoKinetic-ToxicoDynamic, Species Sensitivity Distribution, population, community, and mixture) were searched for into the reports of 317 active substances. At least one model category was found for 44 % of the investigated active substances. The most detected models were Species Sensitivity Distribution, Structure-Activity and ToxicoKinetic for 27, 21 and 15 % of the active substances, respectively. The use of modelling was of particular importance for conventional active substances such as sulfonylurea or carbamates contrary to microorganisms and plant derived substances. This review also highlighted a strong imbalance in model usage among the biological groups considered in the European Regulation (EC) No 1107/2009. For example, models were more often used for aquatic than for terrestrial organisms (e.g., birds, mammals). Finally, a gap between the set of models used in reports and those existing in the literature was observed highlighting the need for the implementation of more sophisticated models into PPP regulation.

How diatom-, invertebrate- and fish-based diagnostic tools can support the ecological assessment of rivers in a multi-pressure context: Temporal trends over the past two decades in France.

The degradation of aquatic ecosystems, induced by worldwide intensification in the use of both land and aquatic resources, has highlighted the critical need for innovative methods allowing an objective quantification and ranking of anthropogenic pressure effects on aquatic organisms. Such diagnostic tools have a great potential for defining robust management responses to anthropogenic pressures. Our objective was to explore how the outputs of three diagnostic tools (based on benthic diatoms, macroinvertebrates and fishes) could be combined to (i) disentangle the temporal effects of multiple pressures over two decades and (ii) provide policy-relevant information for stream managers and decision makers. The diagnostic tools estimated, using taxonomy- and trait-based metrics, the impairment probabilities of biotic assemblages over time by different pressure categories, describing the alteration of water quality, hydromorphology and land use related to anthropogenic activities, in French streams (number of sites = 312). The main result shows that a large proportion of the time series exhibited no significant temporal patterns over the two decades (61.5% to 87.8%, depending on the used tests). Among time series exhibiting significant change, positive trends in impairment probabilities (i.e., degradation) were less frequent than negative ones, indicating a modest improvement in water quality at national scale over the study period. However, trends can be substantially different according to hydroecoregion and pressure category. The three biological compartments displayed convergent temporal responses according to the pressure category and regional context (e.g., lowland plains vs. mountains, pristine vs. agricultural regions). Altogether, this study proposes a unifying approach to integrate a vast amount of information in a single ecological diagnosis using an unparalleled database on natural and anthropized environments. Strengthening the synthesis of biological information provided by various biological compartments should be a priority before implementing evidence-based sustainable conservation and restoration actions.

A multi-omics concentration-response framework uncovers novel understanding of triclosan effects in the chlorophyte Scenedesmus vacuolatus.

In aquatic ecosystems, the biocide triclosan represents a hazard for the non-target microalgae. So far, algal responses were mainly investigated at apical levels hampering the acquisition of a holistic view on primary, adaptive, and compensatory stress responses. We assessed responses of the chlorophyte Scenedesmus vacuolatus to triclosan at apical (growth, photosynthesis) and molecular (transcriptome, metabolome) levels for comparative pathway sensitivity analysis. For each responsive signal (contigs, metabolites), a concentration-response curve was modeled and effect concentrations were calculated leading to the setting of cumulative sensitivity distributions. Molecular responses showed higher sensitivity than apical observations. The functional annotation of contigs and metabolites revealed 118 metabolic pathways putatively impaired by triclosan, highlighting a wide repercussion on the algal metabolism. Metabolites involved in the lipid metabolism showed decreasing trends along the concentration gradient and a globally highest sensitivity, pointing to the primary target of triclosan. The pathways involved in xenobiotic degradation and membrane transporters were mainly regulated in the transcriptome with increasing response trends comprising compensatory responses. The suggested novel approach, combining apical and multi-omics analyses in a concentration-response framework improves mechanistic understanding and mode of action analysis on non-targeted organisms and is suggested to better implement high-throughput multi-omics data in environmental risk assessment.

Assessing anthropogenic pressure in the St. Lawrence River using traits of benthic macroinvertebrates.

This study aims to evaluate the anthropogenic pressure in the St. Lawrence River by assessing the relationships between chemical contamination of sediments and benthic community structure with the trait-based approach. Organic and inorganic contaminants as well as other sediment variables (sediment grain size, total organic carbon, nutrients, etc.) and benthic invertebrate assemblages were determined in 59 sites along the river. Biological and ecological traits of taxa were coded, taking into account regional climate and ecosystem conditions. The aims of this study were to (1) describe the relationships between traits and macroinvertebrate taxa and identify homogeneous clusters of taxa with the same combinations of functional traits, (2) describe spatial patterns in traits of macroinvertebrates in the St. Lawrence River, (3) link trait-based metrics and site groups to sediment quality and (4) define a trait-based strategy for diagnosing the ecological quality of the St. Lawrence River. Seven groups of taxa sharing similar trait-category attributes were defined. Moreover, four groups of sites were identified using the 'K-mean' non-hierarchical clustering approach. The 'IndVal' method enabled us to specifically defined trait categories corresponding to site groups on the basis of their indicator value. The relative abundances of taxa from five functional groups significantly varied among site groups. For example, some indicator traits such as multivoltine cycle, long life span, fixed clutches, tegumental respiration, asexual reproduction, and collector/gatherer feeding habit were associated to the most heavily polluted sites located in the Montreal harbour which showed the highest sediment concentrations in Pb, Zn and Cu. Three trait-based pressure-specific models were built, based on the random forest approach, for respectively (1) heavy metals, (2) BPCs and PAHs, and (3) TBTs occurring in the environment. These models could be applied to assess sediment quality using macroinvertebrate assemblages in a large Canadian river.

Hydrodynamics Alter the Tolerance of Autotrophic Biofilm Communities Toward Herbicides.

Multiple stressors pose potential risk to aquatic ecosystems and are the main reasons for failing ecological quality standards. However, mechanisms how multiple stressors act on aquatic community structure and functioning are poorly understood. This is especially true for two important stressors types, hydrodynamic alterations and toxicants. Here we perform a mesocosm experiment in hydraulic flumes connected as a bypass to a natural stream to test the interactive effects of both factors on natural (inoculated from streams water) biofilms. Biofilms, i.e., the community of autotrophic and heterotrophic microorganisms and their extracellular polymeric substances (EPS) in association with substratum, are key players in stream functioning. We hypothesized (i) that the tolerance of biofilms toward toxicants (the herbicide Prometryn) decreases with increasing hydraulic stress. As EPS is known as an absorber of chemicals, we hypothesize (ii) that the EPS to cell ratio correlates with both hydraulic stress and herbicide tolerance. Tolerance values were derived from concentration-response assays. Both, the herbicide tolerance and the biovolume of the EPS significantly correlated with the turbulent kinetic energy (TKE), while the diversity of diatoms (the dominant group within the stream biofilms) increased with flow velocity. This indicates that the positive effect of TKE on community tolerance was mediated by turbulence-induced changes in the EPS biovolume. This conclusion was supported by a second experiment, showing decreasing effects of the herbicide to a diatom biofilm (Nitzschia palea) with increasing content of artificial EPS. We conclude that increasing hydrodynamic forces in streams result in an increasing tolerance of microbial communities toward chemical pollution by changes in EPS-mediated bioavailability of toxicants.

Assessing anthropogenic pressures on streams: A random forest approach based on benthic diatom communities.

Benthic diatoms have been widely used to assess the ecological status of freshwater ecosystems, especially in the context of recent international water framework directive policies (e.g. the WFD). Despite diatom-based indices are known to respond fastly to water quality degradation, they are not designed to precisely identify the nature of pressures co-occurring in the environment. Based on large scale monitoring data, we aimed at building models able to estimate the risk of stream impairment by many types of anthropogenic pressures from taxonomy-based and trait-based characteristics of diatom assemblages. Random forest models were built to individually evaluate the impairment risk of diatom assemblages for six chemical and five hydromorphological or land-use related pressure categories. Eight models provided good impairment risk assessment (Area Under the Curve≥0.70). Under multi-pressure scenarios, models built for chemical pressures exhibited a better accuracy than hydromorphological or land-use related ones. Models were able to detect both ecological restoration and degradation, based on long-term surveys. These models have been implemented in a R user-friendly routine, to help stream managers to early identify degrading processes and prioritize management actions.

Comparison of specific versus literature species sensitivity distributions for herbicides risk assessment.

Species sensitivity distributions (SSDs) are an important predictive tool in risk assessment. Usually, literature data are used to build SSDs that are mostly based on planktonic species. But, to get adequate protective thresholds for environmental communities, one could argue that SSD should be built on ecotoxicological data obtained from species found in the ecosystem that should be protected. This is particularly true when benthic algae are of concern. Due to the lack of literature data, building SSD on benthic microalgae is difficult. This paper aims in comparing SSDs, and thus protective thresholds (hazardous concentration that affects 5% of the species of a community, HC5), built on ecotoxicological data obtained (1) from literature and (2) with specific bioassays on benthic diatoms from a lake. Thresholds were derived for protection against four herbicides separately and for a mixture of them. Sensitivity data obtained from literature were statistically lower than the specific data for all herbicides: Species tested in the literature were usually more sensitive (mainly chlorophytes), leading to more protective lower HC5. The HC5 thresholds (literature or specific) derived for protection against the mixture were also compared to the observed sensitivity of an assemblage of benthic diatom species exposed to an increasing range of herbicide mixture concentrations. We observed that one species within the assemblage (Fragilaria rumpens) was affected at a concentration below both the literature and the specific HC5 thresholds.

Pollution-induced community tolerance (PICT) as a tool for monitoring Lake Geneva long-term in situ ecotoxic restoration from herbicide contamination.

Chemical monitoring revealed a regular decrease in herbicide concentration in Lake Geneva since last decades that may be linked to an ecotoxic restoration of nontarget phytoplanktonic communities. The Pollution-induced community tolerance (PICT) approach was tested as a tool to monitor the ecotoxic restoration of Lake Geneva for herbicides from 1999 to 2011. We conducted monthly assessments in 1999 and in 2011 for the tolerance of the phytoplankton communities to two herbicides (atrazine and copper), using PICT bioassays. The taxonomical composition of the communities was determined on the same collecting dates. The herbicide concentration decrease during the 12 years significantly influenced the composition of communities. The PICT monitoring indicated that a significant tolerance decrease in the community to both herbicides accompanied the herbicide concentration decrease. PICT measurements for atrazine and copper also changed at the intra-annual level. These variations were mainly due to community composition shifts linked to seasonal phosphorus and temperature changes. PICT monitoring on a seasonal basis is required to monitor the mean tolerance of communities. PICT appeared to be a powerful tool that reflected the toxic effects on environmental communities and to monitor ecotoxic ecosystem restoration.

Assessment of toxicity thresholds in aquatic environments: does benthic growth of diatoms affect their exposure and sensitivity to herbicides?

Benthic diatoms evolved in a biofilm structure, at the interface between water and substrata. Biofilms can adsorb toxicants, such as herbicides, but little is known about the exposure of biofilm organisms, such as benthic diatoms, to these adsorbed herbicides. We assessed the sensitivity of 11 benthic diatoms species to 6 herbicides under both planktonic and benthic conditions using single-species bioassays. The concentration that reduced the growth rate of the population by 10% (EC10) and 50% (EC50), respectively, varied depending on the species, the herbicides, and the growth forms involved. As a general trend, the more hydrophobic the herbicide, the more species were found to be sensitive under benthic growth conditions. Statistical differences (alpha<5%) were observed between the sensitivities under planktonic and benthic growth conditions for many hydrophobic herbicides. A protective effect of the biofilm against herbicides was observed, and this tended to decrease (at both the EC10 and EC50 levels) with increasing hydrophobicity. The biofilm matrix appeared to control exposure to herbicides, and consequently their toxicity towards benthic diatoms. For metolachlor, terbutryn and irgarol, benthic thresholds derived from species sensitivity distributions were more protective than planktonic thresholds. For hydrophobic herbicides, deriving sensitivity thresholds from data obtained under benthic growth seems to offer a promising alternative.

Physiological and proteomic changes suggest an important role of cell walls in the high tolerance to metals of Elodea nuttallii.

Macrophytes bioaccumulate metals, the suggestion being made that they be considered for phytoremediation. However, a thorough understanding of the mechanisms of metal tolerance in these plants is necessary to allow full optimization of this approach. The present study was undertaken to gain insight into Hg and Cd accumulation and their effects in a representative macrophyte, Elodea nuttallii. Exposure to methyl-Hg (23 ng dm(-3)) had no significant effect while inorganic Hg (70 ng dm(-3)) and Cd (281 µg dm(-3)) affected root growth but did not affect shoots growth, photosynthesis, or antioxidant enzymes. Phytochelatins were confirmed as having a role in Cd tolerance in this plant while Hg tolerance seems to rely on different mechanisms. Histology and subcellular distribution revealed a localized increase in lignification, and an increased proportion of metal accumulation in cell wall over time. Proteomics further suggested that E. nuttallii was able to efficiently adapt its energy sources and the structure of its cells during Hg and Cd exposure. Storage in cell walls to protect cellular machinery is certainly predominant at environmental concentrations of metals in this plant resulting in a high tolerance highlighted by the absence of toxicity symptoms in shoots despite the significant accumulation of metals.

Mercury bioaccumulation in the aquatic plant Elodea nuttallii in the field and in microcosm: accumulation in shoots from the water might involve copper transporters.

Previous studies suggest that macrophytes might participate in bioaccumulation and biomagnification of toxic mercury (Hg) in aquatic environment. Hg bioaccumulation and uptake mechanisms in macrophytes need therefore to be studied. Amongst several macrophytes collected in an Hg contaminated reservoir in Romania, Elodea nuttallii showed a high organic and inorganic Hg accumulation and was then further studied in the laboratory. Tolerance and accumulation of Hg of this plant was also high in the microcosm. Basipetal transport of inorganic Hg was predominant, whereas acropetal transport of methyl-Hg was observed with apparently negligible methylation or demethylation in planta. Hg concentrations were higher in roots>leaves>stems and in top>middle>bottom of shoots. In shoots, more than 60% Hg was found intracellularly where it is believed to be highly available to predators. Accumulation in shoots was highly reduced by cold, death and by competition with Cu(+). Hg in E. nuttallii shoots seems to mainly originate from the water column, but methyl-Hg could also be remobilized from the sediments and might drive in part its entry in the food web. At the cellular level, uptake of Hg into the cell sap of shoots seems linked to the metabolism and to copper transporters. The present work highlights an important breakthrough in our understanding of Hg accumulation and biomagnifications: the remobilization of methyl-Hg from sediments to aquatic plants and differences in uptake mechanisms of inorganic and methyl-Hg in a macrophyte.