Water flow velocity disturbances in biofilms are quite well understood, but the consequences for pesticide dissipation have yet to be addressed. We investigated the capacity of biofilms with different histories of hydrological disturbances [frequent flow velocity increase (FI), sporadic flow velocity increase (SI), and a control with constant flow velocity (C)] to dissipate a cocktail composed of herbicide (terbuthylazine), fungicide (tebuconazole), and insecticide (imidacloprid) molecules. Pesticide dissipation and bioaccumulation by biofilms were measured using two different exposure scenarios: i) an open-flow experiment using stream channels and ii) a batch experiment using culture flasks. The effect of flow velocity disturbances on biofilms was characterized by reduced thickness and bacterial density, but higher surface density compared to the constant flow velocity control. The highest dissipation of pesticides was observed in FI biofilms, and the lowest was observed in C biofilms. However, bioaccumulation was significantly higher in SI biofilms regardless of open flow and/or batch experiments. The more hydrophobic molecules (terbuthylazine and tebuconazole) had greater accumulation than imidacloprid in biofilms, and this was irrespective of the hydrological treatment and/or pesticide-exposure approach. Hydrological disturbances enhance pesticide accumulation in biofilms, but further research is needed to understand pesticide remobilization and downstream exposure during these events.

In the context of the ecological sustainability of phosphorus, emerging evidence for the ubiquitous presence of polyphosphate-accumulating bacteria in natural environments invites efforts to reveal their roles in the biogeochemical cycle of phosphorus. This requires high-throughput methods to characterize their structure and dynamics in ecosystems. A promising strategy is to combine the staining of intracellular polyphosphate granules and their subsequent detection by flow cytometry, enabling rapid data acquisition. In this study, we evaluated the potential of this approach by testing various factors that could affect the efficiency and specificity of polyphosphate labeling. Most of our experiments were performed using the 4′,6-diamidino-2-phenylindole dye (DAPI). However, we also carried out a preliminary study using the synthetic fluorochrome JC-D7, a new selective fluorescent dye used for the specific labeling of endogenous polyphosphate in living cells. The assays were performed on Tetrasphaera elongata, a Gram-positive bacterium known to accumulate large amounts of intracellular polyphosphates. We also used six bacterial strains belonging to different phyla, in particular a Gram-negative bacterial strain belonging to the genus Pseudomonas, which is characterized by low levels of cellular polyphosphate. The potential of flow cytometry to quantify and sort polyphosphate-accumulating bacteria in complex environmental samples, including soil, freshwater and sediments, was also examined. Our tests provide useful information for the design of future experiments and highlight the potential pitfalls and limitations of detecting polyphosphate-accumulating bacteria using the cytometric approach. We also show that JC-D7 is a promising dye for achieving these objectives, particularly for enumerating polyphosphate-accumulating bacteria from environmental samples.

Preservation of biodiversity and ecosystem services is critical for sustainable development and human well-being. However, an unprecedented erosion of biodiversity is observed and the use of plant protection products (PPP) has been identified as one of its main causes. In this context, at the request of the French Ministries responsible for the Environment, for Agriculture and for Research, a panel of 46 scientific experts ran a nearly 2-year-long (2020–2022) collective scientific assessment (CSA) of international scientific knowledge relating to the impacts of PPP on biodiversity and ecosystem services. The scope of this CSA covered the terrestrial, atmospheric, freshwater, and marine environments (with the exception of groundwater) in their continuity from the site of PPP application to the ocean, in France and French overseas territories, based on international knowledge produced on or transposable to this type of context (climate, PPP used, biodiversity present, etc.). Here, we provide a brief summary of the CSA’s main conclusions, which were drawn from about 4500 international publications. Our analysis finds that PPP contaminate all environmental matrices, including biota, and cause direct and indirect ecotoxicological effects that unequivocally contribute to the decline of certain biological groups and alter certain ecosystem functions and services. Levers for action to limit PPP-driven pollution and effects on environmental compartments include local measures from plot to landscape scales and regulatory improvements. However, there are still significant gaps in knowledge regarding environmental contamination by PPPs and its effect on biodiversity and ecosystem functions and services. Perspectives and research needs are proposed to address these gaps.

Streams naturally receive allochthonous particulate organic materials, but human activity may contribute additional plastic litter inputs, which can affect ecosystem functioning. Our objective was to assess the effect of plastic and bioplastic inputs on microbial biomass and function in a pristine mountain stream. To do that, fragments of plastics (polyethylene-PE, polypropylene-PP), bioplastics (polyhydroxyalkanoate-PHA, polylactic acid-PLA), and wood (as a natural material) were immersed in the stream and collected after 120, 202, 316, and 383 d. Fungal, prokaryotic, and algal biomass, and autotrophic and heterotrophic functions (net primary production, extracellular enzyme activities, and nutrient uptake) were analyzed together with organic matter weight changes over time. Wood showed high fungal and prokaryotic biomass, phosphatase and beta-glucosidase activities, and a significant weight loss, as related to the degradation process. In contrast, plastics and bioplastics did not lose weight and were mainly colonized by algae, suggesting that they serve as an inert surface and over-enhance primary production. However, phenol oxidase (ligninolytic enzyme) activity in plastics and bioplastics was similar to that in wood and increased with time together with heterotrophic biomass. This indicates that plastispheres can contribute to the degradation of plastics and bioplastics, which may become a carbon source in the long term. Overall, the observed dynamics of microbial biomass and metabolism in the plastispheres point to their potential effect on stream nutrient and carbon cycles. The study highlights the need for careful human activities in pristine mountain watersheds to avoid altering their ecosystem functioning.

Copper-based plant protection products (PPPs) are widely used in both conventional and organic farming, and to a lesser extent for non-agricultural maintenance of gardens, greenspaces, and infrastructures. The use of copper PPPs adds to environmental contamination by this trace element. This paper aims to review the contribution of these PPPs to the contamination of soils and waters by copper in the context of France (which can be extrapolated to most of the European countries), and the resulting impacts on terrestrial and aquatic biodiversity, as well as on ecosystem functions. It was produced in the framework of a collective scientific assessment on the impacts of PPPs on biodiversity and ecosystem services in France. Current science shows that copper, which persists in soils, can partially transfer to adjacent aquatic environments (surface water and sediment) and ultimately to the marine environment. This widespread contamination impacts biodiversity and ecosystem functions, chiefly through its effects on phototrophic and heterotrophic microbial communities, and terrestrial and aquatic invertebrates. Its effects on other biological groups and biotic interactions remain relatively under-documented.

One of the major threats to freshwater biodiversity is water pollution including excessive loads of nutrients, pesticides, industrial chemicals, and/or emerging contaminants. The widespread use of organic pesticides for agricultural and nonagricultural (industry, gardening, etc.) purposes has resulted in the presence of their residues in various environments, including surface waters. However, the contribution of pesticides to the deterioration of freshwater ecosystems (i.e., biodiversity decline and ecosystem functions impairment) remains uncertain. Once in the aquatic environment, pesticides and their metabolites can interact with microbial communities, causing undesirable effects. The existing legislation on ecological quality assessment of water bodies in Europe is based on water chemical quality and biological indicator species (Water Framework Directive, Pesticides Directive), while biological functions are not yet included in monitoring programs. In the present literature review, we analyze 20 years (2000–2020) of research on ecological functions provided by microorganisms in aquatic ecosystems. We describe the set of ecosystem functions investigated in these studies and the range of endpoints used to establish causal relationships between pesticide exposure and microbial responses. We focus on studies addressing the effects of pesticides at environmentally realistic concentrations and at the microbial community level to inform the ecological relevance of the ecotoxicological assessment. Our literature review highlights that most studies were performed using benthic freshwater organisms and that autotrophic and heterotrophic communities are most often studied separately, usually testing the pesticides that target the main microbial component (i.e., herbicides for autotrophs and fungicides for heterotrophs). Overall, most studies demonstrate deleterious impacts on the functions studied, but our review points to the following shortcomings: (1) the nonsystematic analysis of microbial functions supporting aquatic ecosystems functioning, (2) the study of ecosystem functions (i.e., nutrient cycling) via proxies (i.e., potential extracellular enzymatic activity measurements) which are sometimes disconnected from the current ecosystem functions, and (3) the lack of consideration of chronic exposures to assess the impact of, adaptations to, or recovery of aquatic microbial communities from pesticides. Environ Toxicol Chem 2023;42:1867–1888. © 2023 SETAC

The herbicide glyphosate is contaminating a large number of freshwater ecosystems worldwide and its fate and effects remains uncertain in light of the effects of global change. The present study examines how variations in water temperature and light availability relative to global change affect the ability of stream biofilms to degrade the herbicide glyphosate. Biofilms were exposed in microcosms to two levels of water temperature simulating global warming (Ambient = 19-22 degrees C and Warm = 21-24 degrees C) and three levels of light representative of riparian habitat destruction due to land use change (Dark = 0, Intermediate = 600, High = 1200 mu mol photons m- 2 s- 1). Biofilms were acclimated to six different experimental treatments, namely i) ambient temperature without light (AMB_D), ii) ambient temperature and intermediate light (AMB_IL), iii) ambient temperature and high light (AMB_HL), iv) warm temperature without light (WARM_D), v) warm temperature and intermediate light (WARM_IL) and vi) warm temperature and high light (WARM_HL). The ability of biofilms to degrade 50 mu g L-1 of glyphosate was tested. Results showed that water temperature increase, but not light availability increase, significantly increased aminomethyl phosphonic acid (AMPA) production by biofilms. However, the combined increase of temperature and light generated the shortest time to dissipate half of the glyphosate supplied and/or half of the maximum AMPA produced (6.4 and 5.4 days, respectively) by biofilms. Despite light had a major effect in modulating biofilm structural and functional descriptors, the response of certain descriptors (i. e. chlorophyll-a concentration, bacterial density and diversity, nutrient content and PHO activity) to light avail-ability increase depended on water temperature. Specifically, the biofilms in the WARM_HL treatment displayed the highest Glucosidase: Peptidase and Glucosidase: Phosphatase enzyme activity ratios and the lowest biomass C: N molar ratios compared to the other treatments. According to these results, warmer temperatures and high light availability could have been exacerbating the decomposition of organic C compounds in biofilms, including the use of glyphosate as a C source for microbial heterotrophs. This study shows that ecoenzymatic stoichiometry and xenobiotic biodegradation approaches can be combined to better understand the functioning of biofilms in pesticide-polluted streams.

To evaluate the effects of hydrological variability on pesticide dissipation capacity by stream biofilms, we conducted a microcosm study. We exposed biofilms to short and frequent droughts (daily frequency), long and less frequent droughts (weekly frequency) and permanently immersed controls, prior to test their capacities to dissipate a cocktail of pesticides composed of tebuconazole, terbuthylazine, imidacloprid, glyphosate and its metabolite aminomethylphosphonic acid. A range of structural and functional descriptors of biofilms (algal and bacterial biomass, extracellular polymeric matrix (EPS) concentration, microbial respiration, phosphorus uptake and community-level physiological profiles) were measured to assess drought effects. In addition, various parameters were measured to characterise the dynamics of pesticide dissipation by biofilms in the different hydrological treatments (% dissipation, peak asymmetry, bioconcentration factor, among others). Results showed higher pesticide dissipation rates in biofilms exposed to short and frequent droughts, despite of their lower biomass and EPS concentration, compared to biofilms in immersed controls or exposed to long and less frequent droughts. High accumulation of hydrophobic pesticides (tebuconazole and terbuthylazine) was measured in biofilms despite the short exposure time (few minutes) in our open-flow microcosm approach. This research demonstrated the stream biofilms capacity to adsorb hydrophobic pesticides even in stressed drought environments.

Plastic pollution represents a threat for biological communities and the ecological functions they provide in river ecosystems. In this study, we compared the microbial colonization of two plastics (biodegradable and non-biodegradable) and three natural substrata (leaves, sediment, and rocks) in two study sites of an urbanized watershed differing in their plastic-contamination degree (upstream and downstream). The density and diversity of bacterial, fungal, and algal communities, as well as the extracellular enzymatic activities β-glucosidase (GLU), N-acetyl-glucosaminidase (NAG), and phosphatase (PHO), were analysed in each substrata and site over a 4-week colonization experiment. Results showed higher microbial densities and enzymatic activities in leaves and sediment compared to plastics and rocks, probably due to the greater availability of organic carbon and nutrients in the former substrata. However, the microbial colonization of the two plastics was only different in the downstream site, where bacterial density and enzymatic activities were higher in the biodegradable plastic compared to the non-biodegradable plastic. Accordingly, the presence of biodegradable plastics would enhance the heterotrophic metabolism in plastic-polluted rivers.

This article is the result of work presented at the PharmaAQUA seminar (9-11 March, Annecy, France). It contributes towards knowledge on the pollution of inland aquatic ecosystems by pharmaceutical residues (PR) and antibiotic resistant bacteria (ARB). Pollution of these ecosystems by PR occurs around the world. The main sources are effluents from wastewater treatment plants (WWTP) and the spreading of organic waste products of animal origin (manure and slurry). French rivers are no exception and contain PR in concentrations ranging from nanograms per litre to micrograms per litre, depending on the molecule.This chronic contamination generates risks and ecotoxicological effects to which microbial communities are potentially vulnerable. Their alteration can indirectly affect the biodiversity and functioning of ecosystems. The Pollution Induced Community Tolerance (PICT) approach is a promising tool for diagnosing chemical pollution by PR. Aquatic environments are polluted by antibiotic residues and other chemicals that promote the development of a reservoir of antibiotic resistance genes (ARG). Bacterial biofilms and sediments are trapping areas for ARB and ARG. rRecent studies have shown ARB and ARG accumulate downstream of WWTP. Urban and hospital effluents have specific signatures for ARG. The improvement of wastewater treatment by WWTP, the reduction of discharges at source (through information, greater awareness, and changes in practices), and better control of pharmaceutical production are all levers that are being studied to control this pollution. They require multiple and coordinated efforts from all actors involved in this issue, at the intersection of human health, animal health, and water.

Abstract:The connectivity of environmental compartments through chemical and biological exchanges is often difficult to study. However, understanding the functioning of fluxes is essential in the context of climate change and the assessment of anthropogenic impacts. These exchanges can be realized through water cycle fluxes establishing interactions between the atmosphere, surface water and land.To examine the interactions between the atmosphere, surface water and land via water fluxes, we conducted a large scale field study at the watershed level, involving multiple disciplines from chemistry to meteorology and microbiology. The chemical and biological contents of water from the atmosphere (cloud and rain), to mid-mountain hydrological continuum (streams, wetlands and lake) and soil (agricultural plots) were assessed in the natural and agricultural area from Puy De Dôme (Central France) along an altitudinal gradient from puy de Dôme Mountain summit (1465 m asl) to the plain (~ 600 m asl). We set up experimental procedures for sampling, handling and analysing each environmental matrix, and the environmental context was characterized through meteorological and hydrological measurements and models. The biological and chemical variables included: isotopes of water (1H/2H and 16O/18O; laser spectroscopy), major ions (Na+, NH4+, K+, Mg2+, Ca2+, Cl-, NO3-, SO4-, IC), amino acids (LC-MS), bacterial diversity (16S metabarcoding and high-throughput sequencing) and microbial enzymatic activity associated with the nitrogen, carbon and phosphorus cycles (fluorimetric activity assays for whole Beta-Glucosidase, Leucine Aminopeptidase and Phosphatase activities).The multi-compartment analysis revealed significant differences between the compartments by the chemical variables, highlighting the compartment specificity. We estimated a chemical flux of major ions and amino acids from the atmosphere to the surface (soil and surface water). Bacterial diversity analysis showed a core community in these compartments, confirming their connectivity. Thereafter, we tried to explain bacterial diversity by the chemical variables from the studied compartments. Our analysis on microbial enzymatic activity showed an enzymatic activity associated with the nitrogen, carbon and phosphorus cycles in clouds and rain.Here, our study contributes to the understanding of atmosphere-surface interaction through field observations and atmospheric models and we attempted to better understand environmental fluxes. Our field study emphasized the importance of considering the interaction of environmental compartments in future investigations for future and gobal assessments of anthropogenic impacts, such as agrosystem effects to natural ecosystems. Key words:Field observations, Environmental interaction, Chemical flux, Microbial diversity, Atmosphere, Surface water, Soil.

The present study investigates the individual degrading behavior of bacterial strains isolated from glyphosate-degrading stream biofilms. In this aim, biofilms were subjected to enrichment experiments using glyphosate or its metabolite AMPA (aminomethyl phosphonic acid) as the sole phosphorus source. Five bacterial strains were isolated and taxonomically affiliated to Ensifer sp. CNII15, Acidovorax sp. CNI26, Agrobacterium tumefaciens CNI28, Novosphingobium sp. CNI35 and Ochrobactrum pituitosum CNI52. All strains were capable of completely dissipating glyphosate after 125-400 h and AMPA after 30-120 h, except for Ensifer sp. CNII15 that was not able to dissipate glyphosate but entirely dissipated AMPA after 200 h. AMPA dissipation was overall faster than glyphosate dissipation. The five strains degraded AMPA completely since formaldehyde and/or glycine accumulation was observed. During glyphosate degradation, the strain CNI26 used the C-P lyase degradation pathway since sarcosine was quantitatively produced, and C-P lyase gene expression was enhanced 30× compared to the control treatment. However, strains CNI28, CNI35 and CNI52 accumulated both formaldehyde and glycine after glyphosate transformation suggesting that both C-P lyase and/or glyphosate oxidase degradation pathways took place. Our study shows different and complementary glyphosate degradation pathways for bacteria co-existing in stream biofilms.

De nombreux microorganismes tels que les bactéries, les champignons ou la microfaune sont complètement ignorés par la réglementation au sujet de la qualité des masses d’eau. Or tous ces microorganismes jouent un rôle écologique majeur dans les écosystèmes aquatiques. Au cours de la dernière décennie, divers indicateurs microbiens ont démontré leur potentiel pour le diagnostic de la pollution toxique et de ses effets. Après une description de ces outils de diagnostic et leur mise en perspective opérationnelle, cet article explore les freins ayant à ce jour limité leur utilisation.

Freshwater contamination by pesticides in agricultural landscapes is of increasing concern worldwide, with strong pesticide impacts on biodiversity, ecosystem functions, and ultimately human health (drinking water, fishing). In addition, the excessively large number of substances, as well as their low - and temporally variable - concentrations in water, make the chemical monitoring by grab sampling very demanding and not fully representative of the actual contamination. Tools that integrate temporal variations and that are ecologically relevant are clearly needed to improve the monitoring of freshwater contamination and assess its biological effects. Here, we studied pesticide contamination and its biological impacts in 10 stream sections (sites) belonging to 3 agricultural catchments in France. In each site, we deployed a combination of pesticide integrative samplers, biocenotic indicators based on benthic macroinvertebrates, and functional indicators based on leaf litter decomposition and associated fungal communities. The 3 approaches largely proved complementary: structural and functional indicators did not respond equally to different agricultural impacts such as pesticide contamination (as revealed by integrative samplers), nutrients, or oxygen depletion. Combining chemical, structural, and functional indicators thus seems an excellent strategy to provide a comprehensive picture of agricultural impacts on stream ecosystems.

Knowledge on interactions among microbial communities colonizing various streambed substrata (e.g. cobbles, sediment, leaf-litter etc.) is essential when investigating the functioning of stream ecosystems. However, these interactions are often forgotten when assessing the responses of aquatic microbial communities to chemical contamination. Using a stream microcosm approach, the respective impact of two sulfonamide antibiotics (sulfamethoxazole and sulfamethazine) on the ability of microbial heterotrophs to decompose alder leaves was investigated in the presence or absence of periphyton. Our hypothesis suggested that sulfonamides would negatively impair microbial litter decomposition and that periphyton could possibly alleviate this effect by stimulating microbial decomposer activity through a priming effect. Results showed that the presence of periphyton enriched water with oxygen and labile dissolved organic carbon forms. However, these labile organic carbon sources did not stimulate leaf-litter decomposition but mostly decoupled microbial decomposer activity from particulate organic matter to dissolved organic matter through negative priming. Also, the two sulfonamide molecules did not affect the leaf-litter decomposition process but significantly decreased bacterial biomass accrual on leaves. The reduction of bacteria was concomitant with an increase in biomass-specific β-glucosidase activity and this was attributed to a stress response from bacteria to sulfonamides. Further research looking at microbial interactions would provide for better assessment of chemical contamination effects in communities and processes in stream ecosystems.