The present study investigates the effect of metals on the secretion of enzymes from12 fungal strains maintained in liquid cultures. Hydrolases (acid phosphatase, β-glucosidase, β-galactosidase, and N-acetyl-β-glucosaminidase) and ligninolytic oxidoreductases (laccase, Mn, and lignin peroxidases) activities, as well as biomass production, were measured in culture fluids from fungi exposed to Cu or Cd. Our results showed that all fungi secreted most of the selected hydrolases and that about 50 % of them produced a partial oxidative system in the absence of metals. Then, exposure of fungi to metals led to the decrease in biomass production. At the enzymatic level, Cu and Cd modified the secretion profiles of soil fungi. The response of hydrolases to metals was contrasted and complex and depended on metal, enzyme, and fungal strain considered. By contrast, the metals always stimulated the activity of ligninolytic oxidoreductases in fungal strains. In some of them, oxidoreductases were specifically produced following metal exposure. Fungal oxidoreductases provide a more generic response than hydrolases, constituting thus a physiological basis for their use as biomarkers of metal exposure in soils.

International audience | The insecticide chlordecone applied for decades in banana plantations currently contaminates 20,000 ha of arable land in the French West Indies. Although the impact of various pesticides on soil microorganisms has been studied, chlordecone toxicity to the soil microbial community has never been assessed. We investigated in two different soils (sandy loam and silty loam) exposed to different concentrations of CLD (D0, control; D1 and D10, 1 and 10 times the agronomical dose) over different periods of time (3, 7, and 32 days): (i) the fate of chlordecone by measuring C-14-chlordecone mass balance and (ii) the impact of chlordecone on microbial community structure, abundance, and function, using standardized methods (-A-RISA, taxon-specific quantitative PCR (qPCR), and C-14-compounds mineralizing activity). Mineralization of C-14-chlordecone was inferior below 1 % of initial C-14-activity. Less than 2 % of C-14-activity was retrieved from the water-soluble fraction, while most of it remained in the organic-solvent-extractable fraction (75 % of initial C-14-activity). Only 23 % of the remaining C-14-activity was measured in nonextractable fraction. The fate of chlordecone significantly differed between the two soils. The soluble and nonextractable fractions were significantly higher in sandy loam soil than in silty loam soil. All the measured microbiological parameters allowed discriminating statistically the two soils and showed a variation over time. The genetic structure of the bacterial community remained insensitive to chlordecone exposure in silty loam soil. In response to chlordecone exposure, the abundance of Gram-negative bacterial groups (beta-, gamma-Proteobacteria, Planctomycetes, and Bacteroidetes) was significantly modified only in sandy loam soil. The mineralization of C-14-sodium acetate and C-14-2,4-d was insensitive to chlordecone exposure in silty loam soil. However, mineralization of C-14-sodium acetate was significantly reduced in soil microcosms of sandy loam soil exposed to chlordecone as compared to the control (D0). These data show that chlordecone exposure induced changes in microbial community taxonomic composition and function in one of the two soils, suggesting microbial toxicity of this organochlorine.

Passive sampling devices (PS) are widely used for pollutant monitoring in water, but estimation of measurement uncertainties by PS has seldom been undertaken. The aim of this work was to identify key parameters governing PS measurements of metals and their dispersion. We report the results of an in situ intercomparison exercise on diffusive gradient in thin films (DGT) in surface waters. Interlaboratory uncertainties of time-weighted average (TWA) concentrations were satisfactory (from 28% to 112%) given the number of participating laboratories (10) and ultra-trace metal concentrations involved. Data dispersion of TWA concentrations was mainly explained by uncertainties generated during DGT handling and analytical procedure steps. We highlight that DGT handling is critical for metals such as Cd, Cr and Zn, implying that DGT assembly/dismantling should be performed in very clean conditions. Using a unique dataset, we demonstrated that DGT markedly lowered the LOQ in comparison to spot sampling and stressed the need for accurate data calculation.

The effects of polystyrene microbeads (micro-PS; mix of 2 and 6 μm; final concentration: 32 μg L−1) alone or in combination with fluoranthene (30 μg L−1) on marine mussels Mytilus spp. were investigated after 7 days of exposure and 7 days of depuration under controlled laboratory conditions. Overall, fluoranthene was mostly associated to algae Chaetoceros muelleri (partition coefficient Log Kp = 4.8) used as a food source for mussels during the experiment. When micro-PS were added in the system, a fraction of FLU transferred from the algae to the microbeads as suggested by the higher partition coefficient of micro-PS (Log Kp = 6.6), which confirmed a high affinity of fluoranthene for polystyrene microparticles. However, this did not lead to a modification of fluoranthene bioaccumulation in exposed individuals, suggesting that micro-PS had a minor role in transferring fluoranthene to mussels tissues in comparison with waterborne and foodborne exposures. After depuration, a higher fluoranthene concentration was detected in mussels exposed to micro-PS and fluoranthene, as compared to mussels exposed to fluoranthene alone. This may be related to direct effect of micro-PS on detoxification mechanisms, as suggested by a down regulation of a P-glycoprotein involved in pollutant excretion, but other factors such as an impairment of the filtration activity or presence of remaining beads in the gut cannot be excluded. Micro-PS alone led to an increase in hemocyte mortality and triggered substantial modulation of cellular oxidative balance: increase in reactive oxygen species production in hemocytes and enhancement of anti-oxidant and glutathione-related enzymes in mussel tissues. Highest histopathological damages and levels of anti-oxidant markers were observed in mussels exposed to micro-PS together with fluoranthene. Overall these results suggest that under the experimental conditions of our study micro-PS led to direct toxic effects at tissue, cellular and molecular levels, and modulated fluoranthene kinetics and toxicity in marine mussels.

Despite being generally located far from contamination sources, deep marine ecosystems are impacted by chemicals like PCB. The PCB contamination in five fish and shark species collected in the continental slope of the Gulf of Lions (NW Mediterranean Sea) was measured, with a special focus on intra- and interspecific variability and on the driving factors. Significant differences occurred between species. Higher values were measured in Scyliorhinus canicula, Galeus melastomus and Helicolenus dactylopterus and lower values in Phycis blennoides and Lepidorhombus boscii. These differences might be explained by specific abilities to accumulate and eliminate contaminant, mostly through cytochrome P450 pathway. Interindividual variation was also high and no correlation was observed between contamination and length, age or trophic level. Despite its major importance, actual bioaccumulation of PCB in deep fish is not as documented as in other marine ecosystems, calling for a better assessment of the factors driving individual bioaccumulation mechanisms and originating high variability in PCB contamination.

Adding fertiliser to sediments is an established way of studying the effects of eutrophication but a lack of consistent methodology, reporting on enrichment levels, or guidance on application rates precludes rigorous synthesis and meta-analysis. We developed a simple enrichment technique then applied it to 28 sites across an intertidal sandflat. Fertiliser application rates of 150 and 600 g N m−2 resulted in pore water ammonium concentrations respectively 1–110 and 4–580 × ambient, with greater elevations observed in deeper (5–7 cm) than surface (0–2 cm) sediments. These enrichment levels were similar to eutrophic estuaries and were maintained for at least seven weeks. The high between-site variability could be partially explained by the sedimentary environment and macrofaunal community (42%), but only at the high application rate. We suggest future enrichment studies should be conducted in situ across large environmental gradients to incorporate real world complexity and increase generality of conclusions.

International audience | In estuarine ecosystems, metallic and organic contaminants are mainly associated with fine grain sediments which settle on mudflats. Over time, the layers of sediment accumulate and are then transformed by diagenetic processes mainly controlled by microbial activity, recording the history of the estuary's chemical contamination. In an environment of this specific type, we investigated the evolution of the chemical contamination and the structure of both total and active microbial communities, based on PhyloChip analysis of a 4.6-m core corresponding to a 40-year sedimentary record. While the archaeal abundance remained constant along the core, a decrease by one order of magnitude in the bacterial abundance was observed with depth. Both total and active microbial communities were dominated by Proteobacteria, Actinobacteria, and Firmicutes in all sediment samples. Among Proteobacteria, alpha-Proteobacteria dominated both total (from 37 to 60 %) and metabolically active (from 19.7 to 34.6 %) communities, including the Rhizobiales, Rhodobacter, Caulobacterales, and Sphingomonadales orders. Co-inertia analysis revealed a relationship between polycyclic aromatic hydrocarbons, zinc and some polychlorobiphenyls concentrations, and the structure of total and active microbial communities in the oldest and most contaminated sediments (from 1970 to 1975), suggesting that long-term exposure to chemicals shaped the structure of the microbial community.

[Departement_IRSTEA]Ecotechnologies [TR1_IRSTEA]TED | International audience | The German guidelinefor the dimensioning, construction, and operation of constructed wetlands for biological treatment of domestic and municipal wastewater, which last published in March 2006, has recently been revised. The guideline applies to small wastewater treatment systems of less than 50 Person Equivalents (PE), municipal treatment plants up to 1.000 – 3.000 PE (with combined or separated sewers), treatment systems which use constructed wetlands as a polishing step, and small wastewater treatment systems which are operated seasonally (summer months only).The revision is based on a wide range of experience gained in recent years in the use of treatment wetlands in Germany and in Europe. Previous to the revisions, the proposed changes were discussed in a public hearing in Potsdam, Germany in January 2014. The final revisions were released for public review in October 2015. The finalized document is anticipated to be put into effect in April, 2016. A number of constructed wetland designs appear for the first time in the new guideline. Two-stage unsaturated vertical flow gravel filters which receive raw wastewater in the first stage (based on experience in France) as well as two-stage unsaturated vertical flow filters receiving primarytreated wastewater (based on experience in Austria) are now included. The revisions also include new dimensioning for unsaturated vertical flow filters with lava sand as the main treatment media, as well as aerated treatment wetlands, both as secondary treatment steps. Recommendations are also provided for seasonally operated constructed wetlands, as well as constructed wetlands for graywater treatment. With the publication of the new guideline, horizontal subsurface flow constructed wetlands are no longer supported as a secondary treatment stage in Germany, and are recommended only as a tertiary or polishing stage. Existing horizontal subsurface flow wetlands are allowed to continue operation, provided the systems are not hydraulically or organically overloaded, and that they continue to receive routine operations and maintenance. This presentation will outline the key points of the newly revised DWA-A 262 German constructed wetland guideline.

[Departement_IRSTEA]Ecotechnologies [TR1_IRSTEA]TED | International audience | Constructed wetlands for combined sewer overflow treatment (CSO CWs) are variably saturated vertical flow filters in France. The design-support software Orage aims to facilitate engineering work by optimizing filter area and material sitespecifically which was otherwise encumbered by the stochasticity of CSO flows and concentrations. The optimization process relies on measured or simulated CSO series and a low number of input parameters. The iterative shell calls the core model repetitively. During the process, effluent flows and concentrations are simulated from a range of CW domains and compared to legislative thresholds. The iterative shell was tested both with measured and simulated inflows. First, key parameters of the hydraulic optimization were fixed. Large and underscaled designs are excluded this way and succeeding optimizations for pollutant removal are more efficient. Then, the optimization functions were verified using inflow and available land from an existing CSO CW. At third, the automatizations were used to test model predictions in the function of legislative thresholds. Zeolite-enriched media ensures high NH4-N removal at hydraulic loads exceeding the recommendations of present guidelines, marking clogging as an issue for further research. In summary, the demonstrated simulation experiments verified the optimization approach of the dynamic design tool Orage.