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

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

Multi-model assessment of hydrological and environmental impacts on streambed microbes in Mediterranean catchments.

Microbes inhabiting intermittent streambeds are responsible for controlling and developing many biogeochemical processes essential for the ecosystem functions. Although streambed microbiota is adapted to intermittency the intensification of water scarcity and prolonged dry periods may jeopardise their capacity to cope with hydrological changes. This study aims to evaluate whether, and to what extent, the duration of dry periods affects streambed microbial density, diversity, composition (16S rRNA gene diversity) and functions (extracellular enzyme activities and respiration). Our results highlight the fact that hydrology modulates the community composition and, to some extent, the functions carried out under different environmental conditions. The relative abundance of certain taxa inhabiting the driest intermittent communities differs significantly from those found at sites with continuous flow. Microbial functional metrics revealed a progressive increase in recalcitrant carbon degradation activity at sites with an extended dry phase. In contrast, bacterial density and diversity were mainly influenced by the catchment land use, agriculture enhanced density but reduced diversity, and the presence of riparian vegetation supported greater streambed bacterial diversity. From this perspective, a combination of prolonged dryness with reduced riparian vegetation and increased agricultural land cover could compromise the ecosystem functioning by threaten microbially mediated processes linked to the carbon cycle.

The use of copper as plant protection product contributes to environmental contamination and resulting impacts on terrestrial and aquatic biodiversity and ecosystem functions.

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.

Twenty Years of Research in Ecosystem Functions in Aquatic Microbial Ecotoxicology.

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.

Effects of global change on the ability of stream biofilm to dissipate the herbicide glyphosate.

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 °C and Warm = 21-24 °C) and three levels of light representative of riparian habitat destruction due to land use change (Dark = 0, Intermediate = 600, High = 1200 µmol 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 µg 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 availability 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.

Comparison of microbial colonization between natural and plastic substrata in a polluted watershed.

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 ß-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.

Dissipation of pesticides by stream biofilms is influenced by hydrological histories.

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.

Main conclusions and perspectives from the collective scientific assessment of the effects of plant protection products on biodiversity and ecosystem services along the land-sea continuum in France and French overseas territories.

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.

The combination of chemical, structural, and functional indicators to evaluate the anthropogenic impacts on agricultural stream ecosystems.

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.

Glyphosate-degrading behavior of five bacterial strains isolated from stream biofilms.

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.

Effects of sulfonamide antibiotics on aquatic microbial community composition and functions.

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 ß-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.

Interaction between glyphosate and dissolved phosphorus on bacterial and eukaryotic communities from river biofilms.

Since the capacity of river biofilms to degrade glyphosate has been proven to increase when the availability of dissolved phosphorus (P) in water decreases, the present study investigates the diversity responses of bacterial and eukaryotic microbial communities from biofilms in a search for glyphosate-degrader candidates. Glyphosate and P interactions were observed for eukaryotic communities, the highest community richness and diversity being preserved at low concentrations of glyphosate and P. This trend marked by glyphosate was also observed in the structure of eukaryotic communities. Therefore, phosphorus and glyphosate had a synergistic effect in decreasing the richness and diversity of eukaryotes species in biofilms. However, species richness and diversity in bacterial communities were not affected by glyphosate, though shifts in the structure of these communities were concomitant with the degradation of the herbicide. Bacterial communities capable of using glyphosate as P source were characterized by increases in the relative abundance of certain Bacteroidetes, Chloroflexi, Cyanobacteria, Planctomycetes and alpha-Proteobacteria members. Glyphosate-degrader candidates found in natural river biofilms can be further isolated for better understanding of glyphosate degradation pathways, and used as bioremediation strategies in heavily contaminated sites.

Dissolved organic matter does not promote glyphosate degradation in auto-heterotrophic aquatic microbial communities.

Environmental dissolved organic matter (DOM) has been proved to increase microbial population sizes and stimulate the degradation of some pesticide molecules. Among these molecules, the present study investigated the biodegradation of the herbicide glyphosate depending on photoautotrophs DOM supply in a microbial consortium isolated from river biofilms. Degradation experiments in the laboratory were performed in dark and light conditions, as well as after antibiotic supply, in order to characterize the eventual interactions between photoautotrophs and heterotrophs activity during glyphosate degradation. Fifty percent of the initial concentration of glyphosate (0.6 mM) was transformed into aminomethyl phosphonic acid (AMPA) after 9 days in presence or absence of light. Accordingly, the photoautotrophic DOM supply was not stimulating glyphosate degradation by microbial heterotrophs. This lack of response was probably explained by the low net primary production values and weak dissolved organic carbon production recorded in light treatments. The supply of the antibiotic drastically stopped glyphosate transformation demonstrating the central role of bacteria in the biodegradation of the herbicide. Glyphosate also modified the structure of prokaryotes assemblages in the consortium by increasing the relative abundances of Alphaproteobacteria and slightly decreasing those of Gammaproteobacteria. The chemoorganotrophic bacteria Phenylobacterium sp. (Alphaproteobacteria) was related to the transformation of glyphosate in our microbial consortium. The present study highlights the complexity of microbial interactions between photoautotrophs and heterotrophs in microbial assemblages that can contribute to the degradation of pesticides present in aquatic environments.

Stimulation or inhibition: Leaf microbial decomposition in streams subjected to complex chemical contamination.

Leaf litter decomposition is a key mechanism in headwater streams, allowing the transfer of nutrients and energy into the entire food web. However, chemical contamination resulting from human activity may exert a high pressure on the process, possibly threatening the structure of heterotrophic microbial communities and their decomposition abilities. In this study, the rates of microbial Alnus glutinosa (Alnus) leaf decay were assessed in six French watersheds displaying different land use (agricultural, urbanized, forested) and over four seasons (spring, summer, autumn, winter). In addition, for each watershed at each sampling time, both upstream (less-contaminated) and downstream (more-contaminated) sections were monitored. Toxicities (estimated as toxic units) predicted separately for pesticides and pharmaceuticals as well as environmental parameters (including nutrient levels) were related to microbial decay rates corrected for temperature and a range of fungal and bacterial community endpoints, including biomass, structure, and activity (extracellular ligninolytic and cellulolytic enzymatic activities). Results showed that agricultural and urbanized watersheds were more contaminated for nutrients and xenobiotics (higher pesticides and pharmaceuticals predicted toxicity) than forested watersheds. However, Alnus decay rates were higher in agricultural and urbanized watersheds, suggesting compensatory effects of nutrients over xenobiotics. Conversely, fungal biomass in leaves was 2-fold and 1.4-fold smaller in urbanized and agricultural watersheds than in the forested watersheds, respectively, which was mostly related to pesticide toxicity. However, no clear pattern was observed for extracellular enzymatic activities except that ß-glucosidase activity positively correlated with Alnus decay rates. Together, these results highlight microbial communities being more efficient for leaf decomposition in polluted watersheds than in less contaminated ones, which is probably explained by changes in microbial community structure. Overall, our study showed that realistic chemical contamination in stream ecosystems may affect the biomass of Alnus-associated microbial communities but that these communities can adapt themselves to xenobiotics and maintain ecosystem functions.

Meta-analysis of glyphosate contamination in surface waters and dissipation by biofilms.

One consequence of the intensive use of glyphosate is the contamination of rivers by the active substance and its metabolites aminomethyl phosphonic acid (AMPA) and sarcosine, inducing river eutrophication. Biofilms are the predominant lifestyle for microorganisms in rivers, providing pivotal roles in ecosystem functioning and pollutant removal. The persistence of glyphosate in these ecosystems is suspected to be mostly influenced by microbial biodegradation processes. The present study aimed to investigate the tripartite relationship among biofilms, phosphorus and glyphosate in rivers. The first part consists of a co-occurrence analysis among glyphosate, AMPA and phosphorus using an extensive dataset of measurements (n = 56,198) from French surface waters between 2013 and 2017. The second part investigated the capacity of natural river biofilms to dissipate glyphosate, depending on phosphorus availability and the exposure history of the biofilm, in a microcosm study. A strong co-occurrence among glyphosate, AMPA and phosphorus was found in surface waters. More than two-thirds of samples contained phosphorous with glyphosate, AMPA or both compounds. Seasonal fluctuations in glyphosate, AMPA and phosphorus concentrations were correlated, peaking in spring/summer shortly after pesticide spreading. Laboratory experiments revealed that natural river biofilms can degrade glyphosate. However, phosphorus availability negatively influenced the biodegradation of glyphosate and induced the accumulation of AMPA in water. An increase in alkaline phosphatase activity and phosphorus uptake was observed in glyphosate-degrading biofilms, evidencing the tight link between phosphorus limitation and glyphosate degradation by biofilms. The results of the present study show that phosphorus not only is a key driver of river eutrophication but also can reduce complete glyphosate degradation by biofilms and favour the accumulation of AMPA in river water. The predominant role of biofilms and the trophic status of rivers must therefore be considered in order to better assess the fate and persistence of glyphosate.

Flow conditions influence diuron toxicokinetics and toxicodynamics in freshwater biofilms.

Biofilms are considered as good bioindicators of contamination by means of their capacity to react quickly to xenobiotics exposure, and their pivotal role in sustaining the aquatic trophic web. The exchanges of dissolved substances between water column and biofilm can be modulated by flow velocity. This study deals with toxicokinetic (transfer mechanisms) and toxicodynamic (effects) modelling of pesticides under two contrasted flow conditions. Diuron was used to run a 2-h kinetic study on mature biofilms in river channels. Two flow conditions were considered (⋘1 cm·s-1: lentic environments such as ponds, 2 cm·s-1: lotic environments such as watercourses). Three concentrations were tested in order to estimate contamination levels in biofilms: 0, 5 (environmentally relevant concentration) and 50 (to determine the concentration effect) µg·L-1. The effect of the above-mentioned factors was also assessed on biofilms photosynthesis inhibition. For successive sampling times between 0 and 2 h, the raw biofilms and EPS tightly bound to cells plus microorganisms (T-EPS-M), were physically separated and analysed for diuron accumulation and structural and functional microbial descriptors. Diuron amounts accumulated in biofilm increased with increasing diuron exposure. Biofilms accumulated higher amounts of diuron at the lower flow velocity compared to high flow for raw biofilms, while accumulation in the T-EPS-M fraction was similar between flow conditions. Consequently, both flow velocity and diuron exposure had an influence on diuron bioaccumulation and distribution. Photosynthesis inhibition over time was directly linked to the exposure concentration of diuron recorded in the T-EPS-M fraction. These results suggest that flow causes a loss of organic matter in biofilms, decreasing the total accumulation of diuron, especially within diffusible EPS. As pesticide distribution in biofilm is a major factor in the onset of toxicity, the novel fractioning method presented here will improve further toxicokinetic and toxicodynamic studies dealing with biofilms exposed to organic toxicants.

Environmental risk assessment of antibiotics in agroecosystems: ecotoxicological effects on aquatic microbial communities and dissemination of antimicrobial resistances and antibiotic biodegradation potential along the soil-water continuum.

Antibiotics have a wide application range in human and veterinary medicines. Being designed for pharmacological stability, most antibiotics are recalcitrant to biodegradation after ingestion and can be persistent in the environment. Antibiotic residues have been detected as contaminants in various environmental compartments where they cause human and environmental threats, notably with respect to the potential emergence and proliferation of antibiotic-resistant bacteria. An important component of managing environmental risk caused by antibiotics is to understand exposure of soil and water resources to their residues. One challenge is to gain knowledge on the fate of antibiotics in the ecosystem along the soil-water continuum, and on the collateral impact of antibiotics on environmental microorganisms responsible for crucially important ecosystem functions. In this context, the ANTIBIOTOX project aims at studying the environmental fate and impact of two antibiotics of the sulfonamide class of antibiotics, sulfamethazine (SMZ), and sulfamethoxazole (SMX).

Longitudinal development of chlorophyll and phytoplankton assemblages in a regulated large river (the Ebro River).

The distribution of chlorophyll and phytoplankton communities were compared to nutrient concentrations and hydrological parameters along the main stretch of the river Ebro. A progressive increase in planktonic chlorophyll was observed from the upper reaches to the middle section of the river. Chlorophyll reached a maximum (60-80 microg L(-1)) in the meandering section (downstream of the city of Zaragoza), where nutrient inputs (both N and P) and the residence time of the water are very high. In this meandering section phytoplankton assemblages consisted of large centric diatoms and Scenedesmus sp.pl. These longitudinal patterns were interrupted by the presence of three large reservoirs in the lower section of the river. In the section below the reservoirs, the shorter residence water time, the presence of the invasive zebra mussel, and the massive macrophyte development may explain the historical decrease in chlorophyll-a (from 20-45 microg L(-1) in the 1990s to the present 2-5 microg L(-1)). Phytoplankton densities were extremely poor in this section of the river, where large colonial Coelastrum sp.pl. and Pediastrum sp.pl. were the most characteristic taxa.

Interactive Effects of Pesticides and Nutrients on Microbial Communities Responsible of Litter Decomposition in Streams.

Global contamination of streams by a large variety of compounds, such as nutrients and pesticides, may exert a high pressure on aquatic organisms, including microbial communities and their activity of organic matter decomposition. In this study, we assessed the potential interaction between nutrients and a fungicide and herbicide [tebuconazole (TBZ) and S-metolachlor (S-Met), respectively] at realistic environmental concentrations on the structure (biomass, diversity) and decomposition activity of fungal and bacterial communities (leaf decay rates, extracellular enzymatic activities) associated with Alnus glutinosa (Alnus) leaves. A 40-day microcosm experiment was used to combine two nutrient conditions (mesotrophic and eutrophic) with four pesticide treatments at a nominal concentrations of 15 µg L-1 (control, TBZ and S-Met, alone or mixed) following a 2 × 4 full factorial design. We also investigated resulting indirect effects on Gammarus fossarum feeding rates using leaves previously exposed to each of the treatments described above. Results showed interactive effects between nutrients and pesticides, only when nutrient (i.e., nitrogen and phosphorus) concentrations were the highest (eutrophic condition). Specifically, slight decreases in Alnus leaf decomposition rates were observed in channels exposed to TBZ (0.01119 days-1) and S-Met (0.01139 days-1) than in control ones (0.01334 days-1) that can partially be explained by changes in the structure of leaf-associated microbial communities. However, exposition to both TBZ and S-Met in mixture (MIX) led to comparable decay rates to those exposed to the pesticides alone (0.01048 days-1), suggesting no interaction between these two compounds on microbial decomposition. Moreover, stimulation in ligninolytic activities (laccase and phenol oxidase) was observed in presence of the fungicide, possibly highlighting detoxification mechanisms employed by microbes. Such stimulation was not observed for laccase activity exposed to the MIX, suggesting antagonistic interaction of these two compounds on the ability of microbial communities to cope with stress by xenobiotics. Besides, no effects of the treatments were observed on leaf palatability for macroinvertebrates. Overall, the present study highlights that complex interactions between nutrients and xenobiotics in streams and resulting from global change can negatively affect microbial communities associated with leaf litter, although effects on higher trophic-level organisms remains unclear.

Nicosulfuron Degradation by an Ascomycete Fungus Isolated From Submerged Alnus Leaf Litter.

Nicosulfuron is a selective herbicide belonging to the sulfonylurea family, commonly applied on maize crops. Its worldwide use results in widespread presence as a contaminant in surface streams and ground-waters. In this study, we isolated, for the first time, the Plectosphaerella cucumerina AR1 nicosulfuron-degrading fungal strain, a new record from Alnus leaf litter submerged in freshwater. The degradation of nicosulfuron by P. cucumerina AR1 was achieved by a co-metabolism process and followed a first-order model dissipation. Biodegradation kinetics analysis indicated that, in planktonic lifestyle, nicosulfuron degradation by this strain was glucose concentration dependent, with a maximum specific degradation rate of 1 g/L in glucose. When grown on natural substrata (leaf or wood) as the sole carbon sources, the Plectosphaerella cucumerina AR1 developed as a well-established biofilm in 10 days. After addition of nicosulfuron in the medium, the biofilms became thicker, with rising mycelium, after 10 days for leaves and 21 days for wood. Similar biofilm development was observed in the absence of herbicide. These fungal biofilms still conserve the nicosulfuron degradation capacity, using the same pathway as that observed with planktonic lifestyle as evidenced by LC-MS analyses. This pathway involved first the hydrolysis of the nicosulfuron sulfonylurea bridge, leading to the production of two major metabolites: 2-amino-4,6-dimethoxypyrimidine (ADMP) and 2-(aminosulfonyl)-N,N-dimethyl-3-pyridinecarboxamide (ASDM). One minor metabolite, identified as 2-(1-(4,6-dimethoxy-pyrimidin-2-yl)-ureido)-N,N-dimethyl-nicotinamide (N3), derived from the cleavage of the C-S bond of the sulfonylurea bridge and contraction by elimination of sulfur dioxide. A last metabolite (N4), detected in trace amount, was assigned to 2-(4,6-dimethoxy-pyrimidin-2-yl)-N,N-dimethyl-nicotinamide (N4), resulting from the hydrolysis of the N3 urea function. Although fungal growth was unaffected by nicosulfuron, its laccase activity was significantly impaired regardless of lifestyle. Leaf and wood surfaces being good substrata for biofilm development in rivers, P. cucumerina AR1 strain could thus have potential as an efficient candidate for the development of methods aiming to reduce contamination by nicosulfuron in aquatic environments.