Effect of acidification on leaf litter decomposition in benthic and hyporheic zones of woodland streams

Anthropogenic acidification has deleterious effects on both structure and functioning of surface water ecosystems. This study examined how it may affect the leaf decomposition rate and the community structure and activity of decomposers in both benthic and hyporheic zones of five headwater streams along an acidification gradient from highly acidic (pH 4.6) to circumneutral (pH 7.4). Overall, responses to acidification in hyporheic zones were less pronounced, but followed the same pattern as in their benthic counterparts. Leaf decomposition was much faster in the circumneutral stream, both in the hyporheic and benthic zones (k = 0.0068 and 0.0534 d−1, respectively), than in the most acidic one (k = 0.0016 and 0.0055 d−1, respectively), and correlated well with the acidic gradient in both compartments. Interestingly, leaf litter decomposition was less affected by acidification in hyporheic compared to benthic compartments, likely due to the relatively low sensitivity of fungi, which were the main decomposers of buried coarse particulate organic matter. These results argue in favour of conserving hyporheic habitats in acidified streams as they can maintain matter and species fluxes that are essential to the ecosystem

Toxicity of CeO2 nanoparticles on a freshwater experimental trophic chain: A study in environmentally relevant conditions through the use of mesocosms

The toxicity of CeO2 NPs on an experimental freshwater ecosystem was studied in mesocosm, with a focus being placed on the higher trophic level, i.e. the carnivorous amphibian species Pleurodeles waltl. The system comprised species at three trophic levels: (i) bacteria, fungi and diatoms, (ii) Chironomus riparius larvae as primary consumers and (iii) Pleurodeles larvae as secondary consumers. NP contamination consisted of repeated additions of CeO2 NPs over 4 weeks, to obtain a final concentration of 1 mg/L. NPs were found to settle and accumulate in the sediment. No effects were observed on litter decomposition or associated fungal biomass. Changes in bacterial communities were observed from the third week of NP contamination. Morphological changes in CeO2 NPs were observed at the end of the experiment. No toxicity was recorded in chironomids, despite substantial NP accumulation (265.8±14.1mg Ce/kg). Mortality (35.3±6.8%) and a mean Ce concentration of 13.5±3.9mg/kg were reported for Pleurodeles. Parallel experiments were performed on Pleurodeles to determine toxicity pathways: no toxicity was observed by direct or dietary exposures, although Ce concentrations almost reached 100 mg/kg. In view of these results, various toxicity mechanisms are proposed and discussed. The toxicity observed on Pleurodeles in mesocosm may be indirect, due to microorganism’s interaction with CeO2 NPs, or NP dissolution could have occurred in mesocosm due to the structural complexity of the biological environment, resulting in toxicity to Pleurodeles. This study strongly supports the importance of ecotoxicological assessment of NPs under environmentally relevant conditions, using complex biological systems

Decoding the Time-Dependent Response of Bioluminescent Metal-Detecting Whole-Cell Bacterial Sensors

International audienceThe signal produced by aqueous dispersions of bioluminescent, metal-responsive whole-cell bacterial sensors is indicative of the concentration of bioavailable metal ions in solution. The conventional calibration-based strategy followed for measuring this concentration is however inadequate to provide any quantitative prediction of the cells response over time as a function of e.g. their growth features, their defining metal accumulation properties, or the physicochemical medium composition. Such an evaluation is still critically needed for assessing on a mechanistic level the performance of biosensors in terms of metal bioavailability and toxicity monitoring. Herein we report a comprehensive formalism unraveling how the dependence of bioluminescence on time is governed by the dynamics of metal biouptake, by the activation kinetics of lux-based reporter gene, by the ensuing rate of luciferase production, the kinetics of light emission and quenching. It is shown that bioluminescence signal corresponds to the convolution product between two time-dependent functions, one detailing the dynamic interplay of the above micro-and nanoscale processes, and the other pertaining to the change in concentration of photoactive cell sensors over time. Numerical computations illustrate how the shape and magnitude of the bioluminescence peak(s) are intimately connected to the dependence of the photoactive cells concentration on time and to the magnitudes of Deborah numbers that compare the relevant timescales of the biointerfacial and intracellular events controlling light emission. Explicit analytical expressions are further derived for practical situations where bioluminescence is proportional to the concentration of metal ions in solution. The theory is further quantitatively supported by experiments performed on luminescent cadmium-responsive lux-based Escherichia coli biosensors

Bimodal stringence-mediated response of metal-detecting luminescent whole cell bioreporters: experimental evidence and quantitative theory

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Impact of manufactured TiO2 nanoparticles on planktonic and sessile bacterial communities

International audienceIn the present study, we conducted a 2 week microcosm experiment with a natural freshwater bacterial community to assess the effects of titanium dioxide nanoparticles (TiO2-NPs) at various concentrations (0, 1, 10 and 100 mg/L) on planktonic and sessile bacteria under dark conditions. Results showed an increase of planktonic bacterial abundance at the highest TiO2-NP concentration, concomitant with a decrease from that of sessile bacteria. Bacterial assemblages were most affected by the 100 mg/L TiO2-NP exposure and overall diversity was found to be lower for planktonic bacteria and higher for sessile bacteria at this concentration. In both compartments, a 100 mg/L TiO2-NPs exposure induced a decrease in the ratio between the Betaproteobacteria and Bacteroidetes. For planktonic communities, a decrease of Comamonadaceae was observed concomitant with an increase of Oxalobacteraceae and Cytophagaceae (especially Emticicia). For sessile communities, results showed a strong decrease of Betaproteobacteria and particularly of Comamonadaceae

Insight into the primary mode of action of TiO2 nanoparticles on Escherichia coli in the dark.

16 pagesInternational audienceLarge-scale production and incorporation of titanium dioxide nanoparticles (NP-TiO2 ) in consumer products leads to their potential release into the environment and raises the question of their toxicity. The bactericidal mechanism of NP-TiO2 under UV light is known to involve oxidative stress due to the generation of reactive oxygen species. In the dark, several studies revealed that NP-TiO2 can exert toxicological effects. However, the mode of action of these nanoparticles is still controversial. In the present study, we used a combination of fluorescent probes to show that NP-TiO2 causes Escherichia coli membrane depolarization and loss of integrity, leading to higher cell permeability. Using both transcriptomic and proteomic global approaches we showed that this phenomenon translates into a cellular response to osmotic stress, metabolism of cell envelope components and uptake/metabolism of endogenous and exogenous compounds. This primary mechanism of bacterial NP-TiO2 toxicity is supported by the observed massive cell leakage of K(+) /Mg(2+) concomitant with the entrance of extracellular Na(+) , and by the depletion of intracellular ATP level

Modifications of the bacterial reverse mutation test reveals mutagenicity of TiO2 nanoparticles and byproducts from a sunscreen TiO2-based nanocomposite

International audienceThe bacterial reverse mutation test, recommended by the Organization for Economic Co-operation and Development (OECD) to determine genotoxicity of chemical compounds, has been recently used by several authors to investigate nanoparticles. Surprisingly, test results have been negative, whereas in vitro mammalian cell tests often give positive genotoxic responses. In the present study, we used the fluctuation test procedure with the Salmonella typhimurium strains TA97a, TA98, TA100 and TA102 to determine the mutagenic potential of TiO2 nanoparticles (NP-TiO2) and showed that, when it is used conventionally, this test is not suitable for nanoparticle genotoxicity assessment. Indeed, the medium used during exposure prevents electrostatic interactions between bacterial cells and nanoparticles, leading to false-negative responses. We showed that a simple pre-exposure of bacteria to NP-TiO2 in a low ionic strength solution (NaCl 10 mM) at a pH below the nanoparticle isoelectric points (pH 5.5) can strongly improve the accuracy of the test. Thus, based on these improvements, we have demonstrated the genotoxicity of the engineered NP-TiO2 tested and a NP-TiO2 byproduct from a sunscreen nanocomposite. It was also shown that strain TA102 is more sensitive than the other strains, suggesting an oxidative stress-mediated mechanism of genotoxicity. (C) 2012 Elsevier Ireland Ltd. All rights reserved

What do luminescent bacterial metal-sensors probe? Insights from confrontation between experiments and flux-based theory

International audienceWhole-cell bioreporters are routinely operated as sentinels for monitoring the concentration of bioavailable and/or toxic metal ions (M) in aquatic media. Despite the importance of metal bioreporters in environmental risk assessment, their use is often limited to the establishment and exploitation of calibration curves relating bioreporters signal and target metal concentration. In this work, a physicochemical rationale is elaborated for the response of metal-sensitive whole-cell bioreporters beyond the restrictive representation of metal partitioning equilibrium at the microorganism-solution interface. The analysis is conducted under poorly metal complexing conditions for steady-state bioreporter functioning defined by a rate of photons production independent of time. The theoretical framework deciphers how this rate is determined by (i) metal biouptake dynamics with contributions from metal conductive diffusion to the cell surface and metal internalisation kinetics, (ii) formation kinetics and stability of intracellular complexes between M and transcriptional regulators, and (iii) the flux of emitted photons resulting from biochemical reactions initiated after activation of transcriptional regulators. The formalism enables quantitative evaluation of bioreporters performance depending on interfacial cell electrostatics, cell concentration and cell metal-adsorption features. The theory is supported by experimental data on cadmium detection by genetically modified luminescent Escherichia coli bioreporters exhibiting various lipopolysaccharidic surface structures

Kinetics of metal detection by luminescence-based whole-cell biosensors: connecting biosensor response to metal bioavailability, speciation and cell metabolism

International audienceLuminescent whole-cell metal biosensors are genetically engineered cells used for the detection of metals in e.g. aqueous solutions. Herein, we detail the quantitative connections between time-response of luminescent bacterial metal sensors and the bioavailability of free and complexed metal species. To that end, we formulate the biophysicochemical dynamics of metal partitioning at a biosensor/solution interface and integrate the required metabolism contribution to cell response. The formalism explains the ways in which cell signal depends on: coupled Eigen kinetics of metal complexation and diffusion of metal species to/from the interface; kinetics of metal excretion, Michaelis–Menten bioaccumulation and ensuing metal depletion from bulk solution; and kinetics of bioluminescence production following intracellular metal sequestration by regulatory metalloproteins. In turn, an expression is derived for the time-dependent cell signal as a function of interrelated (bioavai)lability of metal species and (thermo)dynamic descriptors of extra/intracellular metal complexation. Quantitative criteria are elaborated to identify scenarios where equilibrium modeling of metal speciation is incorrect, bulk metal depletion is operative, metal biouptake kinetics is governed by metal diffusion, or labile metal complexes fully contribute to cell response. Remarkably, in agreement with experiments, the theory predicts time-shifts of bioluminescence peaks with increasing concentration of biosensor and/or metal ligand in solution. We show that these shifts originate from the crosstalk between activation kinetics of cell photoactivity and speciation-dependent kinetics of bulk metal depletion. Overall, the work paves the way for the elaboration of new strategies to exploit the bioluminescence response of metal lux-biosensors at a dynamic level and evaluate metal bioavailability properties in environmental or biological aqueous samples