Monitored natural attenuation of a complex mixture of organochlorines using compound-specific isotope analysis.

International audience(Calibri, 12pt bold) In France, chlorinated hydrocarbons represent the third most commonly detected groundwater contaminant class (19%, directly after heavy metals, 22%, and mineral‐oil hydrocarbons, 33%; Medde, 2012). The fate and behaviour of organic contaminants in aquifer systems depend on a number of physicochemical and biological processes, which may lead to natural attenuation. To apply in situ attenuation by naturally occurring or enhanced biodegradation as a cost‐effective remediation option at these sites, it becomes primordial to accurately identify and quantify ongoing biotransformation processes. Determination and quantification of these processes are crucial for contaminated site risk assessment and sustainable groundwater management strategies. Compound‐specific isotope analysis (CSIA) by online‐coupling of gas chromatography and isotope ratio mass spectrometry (GC‐IRMS) offers a versatile tool to study the origin of a pollution and/or to decipher multiple contamination sources, to verify if biodegradation of organic pollutants is occurring, to identify degradation mechanisms, and to determine the rate and extent of in‐situ biodegradation. In the last decade, compound‐specific isotope analysis (CSIA) has evolved as a diagnostic tool for contaminated site investigation and for monitoring the efficiency of intrinsic natural or enhanced remediation processes, and is now available for studying an increasing number of environmental pollutants (Elsner et al. 2012). Chemical compounds contain heavy and light stable isotopes in a certain ratio, i.e. 13 C/ 12 C, D/H, and 15 N/ 14 N. In general, molecules containing the lighter isotope react faster than those containing the heavier isotope. Bond‐breaking processes induced by microbial metabolism, can cause a large isotope fractionation leading to an enrichment of heavy isotopes in the residual fraction of the contaminant. CSIA allows interpreting these changes in the isotopic composition of individual organic compounds, thus providing a direct proxy for chemical transformations. CSIA permits qualitative and quantitative information on in situ (bio)degradation, can distinguish different reaction mechanisms, and provides additional means for source allocation and differentiation (Schmidt et al. 2004, Blessing et al. 2009, Braeckevelt et al. 2012, Negrel et al. 2012). We applied compound‐specific carbon isotope analysis (CSIA) in a remediation study to evaluate the potential of monitored natural attenuation (MNA) at an industrial site. The study site is an organochlorine production site with a long operational history and leaking facilities or accidental spills released high amounts of contaminants to soil and groundwater. A control and monitoring program of DNAPL at the study site is installed since the 80s. A hydraulical confinement separated the DNAPL source from the contamination plume that consists of lighter, more soluble chlorinated compounds, mainly PCE, TCE, DCE, TeCA, DCA, TCP and DCP. Biostimulation using acetate and H2 amendment was performed in a limited area within the plume. The aim of the present work is to demonstrate the potential of the CSIA approach for monitoring in‐situ biodegradation and for characterizing and quantifying transformation processes of chlorinated pollutants in groundwater towards the ultimate goal of an implementation of natural and/or enhanced bioremediation at the contaminated site. To this end, a total of four monitoring wells in the source zones and nine groundwater wells within the downgradient contaminant plume over several kilometres length were sampled and stable carbon isotope ratios of the aforementioned chlorinated compounds wer

Analysis of biofilm-nanoparticles interaction using microscopy (fluorescence, MEB, STEM, MET, EDS)

International audienceAmong biofilm's properties, the ability to interact with/catch pollutants can have applications in bioremediation. Here, biofilm interactions with metals (as iron nanoparticles (NanoFer 25S)) was evaluated using various approaches in microscopy. For this, biofilm growth, sampling, labelling and treatment were developed for each type of microscopy to access the surface or inside of the biofilm, biofilm composition, and metal location. Multispecies biofilms were grown on sand or in PVC tubes inoculated with aquifer water spiked with a nutritive solution to enhance denitrification, and then put in contact with nanoparticles. According to the targeted microscopy, biofilms were (i) sampled as flocs or attached biofilm, (ii) submitted to cells (DAPI) and/or lectins (PNA and ConA coupled to FITC or Au nanoparticles) labelling, and (iii) prepared for observation (fixation, cross-section, freezing…). Fluorescent microscopy revealed that nanoparticles were embedded in the biofilm structure as 0.5-5µm size aggregates. SEM observations also showed NP aggregates closed to microorganisms but it was not possible to conclude a potential interaction between nanoparticles and the biological membranes. STEM-in-SEM analysis showed NP aggregates could enter inside the biofilm over a depth of 7-11µm. Moreover, microorganisms were circled by an EPS ring that prevented the direct interaction between NP and membrane. TEM(STEM)/EDS revealed that NP aggregates were co-localized with lectins suggesting a potential role of exopolysaccharides in NP embedding. The combination of several approaches in microscopy is thus a good tool to better understandi and characterize biofilm/pollutant interaction

Regional distributon of mercury in sediments of the main rivers of French Guiana

International audienceUse of mercury (Hg) for gold-mining in French Guiana (up until 2006) as well as the presence of naturally high background levels in soils, has led to locally high concentrations in soils and sediments. The present study maps the levels of Hg concentrations in river sediments from five main rivers of French Guiana (Approuague River, Comté River, Mana River, Maroni River and Oyapock River) and their tributaries, covering more than 5 450 km of river with 1 211 sampling points. The maximum geological background Hg concentration, estimated from 241 non-gold-mined streams across French Guiana was 150 ng g-1. Significant differences were measured between the five main rivers as well as between all gold-mining and pristine areas, giving representative data of the Hg increase due to past gold-mining activities. These results give a unique large scale vision of Hg contamination in river sediments of French Guiana and provide fundamental data on Hg distribution in pristine and gold-mined areas

Shift in Natural Groundwater Bacterial Community Structure Due to Zero-Valent Iron Nanoparticles (nZVI)

Toxic and persistent contaminants in groundwater are technologically difficult to remediate. Remediation techniques using nanoparticles (NPs) such as nZVI (Zero-Valent Iron) are applicable as in situ reduction or oxidation agents and give promising results for groundwater treatment. However, these NP may also represent an additional contamination in groundwater. The aims of this study are to assess the impact of nZVI on the nitrate-reducing potential, the abundance and the structure of a planktonic nitrate-reducing bacterial community sampled in groundwater from a multicontaminated site. An active nitrate-reducing bacterial community was obtained from groundwater samples, and inoculated into batch reactors containing a carbon substrate, nitrate and a range of nZVI concentrations (from 0 to 70.1 mg Fe.L-1). Physical (pH, redox potential), chemical (NO3− concentrations) and biological (DNA, RNA) parameters were monitored during 1 week, as well as nZVI size distribution and mortality of bacteria. Nitrate-reducing activity was temporally stopped in the presence of nZVI at concentrations higher than 30 mg L-1, and bacterial molecular parameters all decreased before resuming to initial values 48 h after nZVI addition. Bacterial community composition was also modified in all cultures exposed to nZVI as shown by CE-SSCP fingerprints. Surprisingly, it appeared overall that bacteria viability was lower for lower nZVI concentrations. This is possibly due to the presence of larger, less reactive NP aggregates for higher nZVI concentrations, which inhibit bacterial activity but could limit cell mortality. After 1 week, the bacterial cultures were transplanted into fresh media without nZVI, to assess their resilience in terms of activity. A lag-phase, corresponding to an adaptation phase of the community, was observed during this step before nitrate reduction reiterated, demonstrating the community’s resilience. The induction by nZVI of modifications in the bacterial community composition and thus in its metabolic potentials, if also occurring on site, could affect groundwater functioning on the long term following nZVI application. Further work dedicated to the study of nZVI impact on bacterial community directly on site is needed to assess a potential impact on groundwater functioning following nZVI application

Impact de nanoparticules de fer zéro-valent (nZVI) sur la structure de la communauté bactérienne issue d'une eau souterraine

International audienc

Impact d'un traitement au H2O2 sur la biodégradation des BTEX évaluée par fractionnement isotopique du carbone stable et abondance des gènes codant des enzymes spécifiques à la dégradation des BTEX

International audienc

Étude des interactions entre sols-mercure-composante microbiologique en Guyane française

Les teneurs élevées en mercure des sols Guyanais suscitent beaucoup d’attention de par les conséquences écotoxicologiques qui en découlent. Dans cet écosystème tropical particulier, notre recherche a eu pour objectifs i) d’évaluer l’importance des oxydes de fer dans la fixation du mercure ii) d’étudier l’impact des processus biogéochimiques dans la remobilisation de ce métal et iii) d’évaluer l’impact de ce mercure mobile sur le fonctionnement et la diversité génétique des communautés microbiennes du sol. Afin d’atteindre ces objectifs deux volets ont été abordés, avec comme matériel d’étude des sols naturels provenant d’une toposéquence proche d’un ancien site orpaillé, situé dans les environs du village de Cacao, et des sols jamais orpaillés de la forêt de Patagai. Dans le premier volet, les résultats obtenus ont montré que les quantités de mercure dans les sols étudiés diminuent le long de la toposéquence, des oxisols aux sols hydromorphes, de même que les teneurs en fer et ceci en étroite relation avec les conditions réductrices et les mouvements probables des nappes d’eau. Les analyses pédologiques combinées à des extractions chimiques sélectives des formes d’oxydes de fer ont permis de montrer l’importance de ces derniers et de leurs propriétés cristallochimiques dans la fixation du mercure. Ces résultats ont logiquement ouvert la voie à une étude microbiologique, visant à évaluer la capacité des bactéries ferri-réductrices à mobiliser le mercure, dans des conditions physico-chimiques réductrices proches de celles rencontrées en conditions naturelles. Menée en microcosme au laboratoire et en présence d’une source de carbone facilement biodégradable, cette étude a mis en évidence une redistribution du mercure parallèlement à l’activité ferri-réductrice bactérienne. Le rôle méthylateur de ces bactéries expliquerait les fortes concentrations de methylmercure (MeHg) rapportées dans les sols inondés de notre site d’étude. Le second volet de ce travail a été consacré à l’impact d’un apport du mercure sur les communautés microbiennes telluriques. Ce volet a été abordé à travers une approche multiscalaire combinant chimie, microbiologie, enzymologie et biologie moléculaire. Les résultats obtenus ont confirmé que l’effet du mercure dépend des teneurs appliquées. Alors que la densité microbienne ne semble pas modifiée, les diversités spécifiques, fonctionnelles ainsi que les cinétiques de minéralisation de carbone sont rapidement influencées par des fortes concentrations en mercure (20 µg/g de sol). D’autre part, si l’application du mercure a montré une modification irréversible à long terme de la structure génétique des communautés microbiennes, un phénomène de résilience a été rapidement constaté au niveau de la minéralisation du carbone et de la diversité fonctionnelle. Un travail complémentaire a permis de mettre en place une méthode d’évaluation multidimensionnelle de la résilience des communautés microbiennes à un stress environnementalGuyanese soils contain high concentrations of mercury due to high geological background concentrations and locally to past gold-mining. The dynamics of this mercury is actually a major environmental and health preoccupation. In this particular ecosystem, the objectives of this thesis were to determine i) the importance of iron oxides in mercury distribution in soils, ii) the impact of microbial ferrireducing bacteria on the remobilisation of this toxic heavy metal, and iii) the impact of mercury pollution on the activity and genetic structure of soil microbial communities. This was assessed through two approaches, the first using soils sampled along a natural ferrallitic toposequence of a catchment basin, partly gold-mined up to 1950, near the village of Cacao. And the second using soils sampled from the forest of Patagai that records no past gold mining. Our results showed that the quantities of mercury decreased along the toposequence along with total iron contents, from the well drained oxisols to the hydromorphic talwegs. This happened in close relation to reductive conditions and watershed dynamics. The soil analyses combined to chemical extractions of amorphous and crystalline iron forms revealed the importance of the latter in mercury adsorption. These results logically brought us to study the impact of microbial activity, and more precisely bacterial iron reduction, on the remobilisation of mercury in these soils. This was carried out in reductive conditions similar to those encountered in natural conditions. In presence of available carbon, these experiments showed that ferri-reducing activity could solubilise significant quantities of iron, thus simultaneously mobilizing mercury. However, we did not detect an increase in dissolved mercury, presumably because it was re precipitated as HgS. Never the less, we observed variations in the amount of mercury associated to iron oxides during incubation. The second part of this work studied the impact of mercury on soil microbial communities. We approached this through a multidisciplinary study including chemistry, microbiology and molecular biology. Results confirmed that the effect of mercury depends on the concentration of Hg applied. While microbial biomass and numbers did not seem to be affected, the genetic structure as well as the functional diversity and carbon mineralisation were rapidly affected by high mercury concentrations (20 µg g-1 of soil). More over, we observed a rapid resilience in carbon mineralisation and functional diversity whereas genetic structure was durably modified. Complementary work enabled us to set up a multidimensional method to evaluate microbial community’s resistance and resilience to an environmental stres

Degradation of tetrachloroethylene by zero valent iron nanoparticles in the presence of a natural groundwater bacterial biofilm in a sandy porous media

International audienceBiofilms are naturally present in aquifers and can interact with zero valent iron nanoparticles (nZVI) used as remediation agents in contaminated groundwater; thereby they may alter nZVI reactivity towards targeted contaminants in porous media. Laboratory scale experiments using columns filled with sand (50 cm long and 5.2 cm in diameter) were performed to investigate the impact of natural biofilms on nZVI reactivity towards tetrachloroethylene (PCE) in conditions simulating an unconsolidated sandy aquifer. Solutions containing PCE were injected through the sand columns in the presence or absence of biofilm and nZVI. Concentrations in PCE and its metabolites were monitored during 45 days in dissolved and gas phases. PCE concentrations decreased at the column outlets due both to its reductive dechlorination by nZVI (~30% of injected PCE) and its sorption or deposition (as PCE-DNAPL) on sand (~35% of injected PCE). No significant differences in PCE concentrations were found in presence or absence of biofilm. However, biofilm presence affected the nature of PCE metabolites. A higher release of ethene in the column containing biofilm was observed, whereas ethane was dominant in the absence of biofilm. Microbes consumed H 2 released by the corrosion of nZVI limiting the hydrogenation of ethene to ethane. The consequences of biofilm development in porous media should be taken into account when considering treatment with nZVI, as it may affect the nature of produced metabolites

Étude des interactions entre sols-mercure-composante microbiologique en Guyane française

Les teneurs élevées en mercure des sols Guyanais suscitent beaucoup d attention de par les conséquences écotoxicologiques qui en découlent. Dans cet écosystème tropical particulier, notre recherche a eu pour objectifs i) d évaluer l importance des oxydes de fer dans la fixation du mercure ii) d étudier l impact des processus biogéochimiques dans la remobilisation de ce métal et iii) d évaluer l impact de ce mercure mobile sur le fonctionnement et la diversité génétique des communautés microbiennes du sol. Afin d atteindre ces objectifs deux volets ont été abordés, avec comme matériel d étude des sols naturels provenant d une toposéquence proche d un ancien site orpaillé, situé dans les environs du village de Cacao, et des sols jamais orpaillés de la forêt de Patagai. Dans le premier volet, les résultats obtenus ont montré que les quantités de mercure dans les sols étudiés diminuent le long de la toposéquence, des oxisols aux sols hydromorphes, de même que les teneurs en fer et ceci en étroite relation avec les conditions réductrices et les mouvements probables des nappes d eau. Les analyses pédologiques combinées à des extractions chimiques sélectives des formes d oxydes de fer ont permis de montrer l importance de ces derniers et de leurs propriétés cristallochimiques dans la fixation du mercure. Ces résultats ont logiquement ouvert la voie à une étude microbiologique, visant à évaluer la capacité des bactéries ferri-réductrices à mobiliser le mercure, dans des conditions physico-chimiques réductrices proches de celles rencontrées en conditions naturelles. Menée en microcosme au laboratoire et en présence d une source de carbone facilement biodégradable, cette étude a mis en évidence une redistribution du mercure parallèlement à l activité ferri-réductrice bactérienne. Le rôle méthylateur de ces bactéries expliquerait les fortes concentrations de methylmercure (MeHg) rapportées dans les sols inondés de notre site d étude. Le second volet de ce travail a été consacré à l impact d un apport du mercure sur les communautés microbiennes telluriques. Ce volet a été abordé à travers une approche multiscalaire combinant chimie, microbiologie, enzymologie et biologie moléculaire. Les résultats obtenus ont confirmé que l effet du mercure dépend des teneurs appliquées. Alors que la densité microbienne ne semble pas modifiée, les diversités spécifiques, fonctionnelles ainsi que les cinétiques de minéralisation de carbone sont rapidement influencées par des fortes concentrations en mercure (20 g/g de sol). D autre part, si l application du mercure a montré une modification irréversible à long terme de la structure génétique des communautés microbiennes, un phénomène de résilience a été rapidement constaté au niveau de la minéralisation du carbone et de la diversité fonctionnelle. Un travail complémentaire a permis de mettre en place une méthode d évaluation multidimensionnelle de la résilience des communautés microbiennes à un stress environnementalGuyanese soils contain high concentrations of mercury due to high geological background concentrations and locally to past gold-mining. The dynamics of this mercury is actually a major environmental and health preoccupation. In this particular ecosystem, the objectives of this thesis were to determine i) the importance of iron oxides in mercury distribution in soils, ii) the impact of microbial ferrireducing bacteria on the remobilisation of this toxic heavy metal, and iii) the impact of mercury pollution on the activity and genetic structure of soil microbial communities. This was assessed through two approaches, the first using soils sampled along a natural ferrallitic toposequence of a catchment basin, partly gold-mined up to 1950, near the village of Cacao. And the second using soils sampled from the forest of Patagai that records no past gold mining. Our results showed that the quantities of mercury decreased along the toposequence along with total iron contents, from the well drained oxisols to the hydromorphic talwegs. This happened in close relation to reductive conditions and watershed dynamics. The soil analyses combined to chemical extractions of amorphous and crystalline iron forms revealed the importance of the latter in mercury adsorption. These results logically brought us to study the impact of microbial activity, and more precisely bacterial iron reduction, on the remobilisation of mercury in these soils. This was carried out in reductive conditions similar to those encountered in natural conditions. In presence of available carbon, these experiments showed that ferri-reducing activity could solubilise significant quantities of iron, thus simultaneously mobilizing mercury. However, we did not detect an increase in dissolved mercury, presumably because it was re precipitated as HgS. Never the less, we observed variations in the amount of mercury associated to iron oxides during incubation. The second part of this work studied the impact of mercury on soil microbial communities. We approached this through a multidisciplinary study including chemistry, microbiology and molecular biology. Results confirmed that the effect of mercury depends on the concentration of Hg applied. While microbial biomass and numbers did not seem to be affected, the genetic structure as well as the functional diversity and carbon mineralisation were rapidly affected by high mercury concentrations (20 g g-1 of soil). More over, we observed a rapid resilience in carbon mineralisation and functional diversity whereas genetic structure was durably modified. Complementary work enabled us to set up a multidimensional method to evaluate microbial community s resistance and resilience to an environmental stressPARIS-EST-Université (770839901) / SudocSudocFranceF

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron (Oxy)Hydroxides, Trace Elements, and Bacteria

International audienceFate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as redox potential (Eh) and pH, but also to microbial activities that can play a direct or indirect role on speciation and/or mobility. Indeed, some bacteria can directly oxidize As(III) to As(V) or reduce As(V) to As(III). Likewise, bacteria are strongly involved in Hg cycling, either through its methylation, forming the neurotoxin monomethyl mercury, or through its reduction to elemental Hg degrees. The fates of both As and Hg are also strongly linked to soil or aquifer composition; indeed, As and Hg can bind to organic compounds or (oxy) hydroxides, which will influence their mobility. In turn, bacterial activities such as iron (oxy) hydroxide reduction or organic matter mineralization can indirectly influence As and Hg sequestration. The presence of sulfate/sulfide can also strongly impact these particular elements through the formation of complexes such as thio-arsenates with As or metacinnabar with Hg.Consequently, many important questions have been raised on the fate and speciation of As and Hg in the environment and how to limit their toxicity. However, due to their reactivity towards aquifer components, it is difficult to clearly dissociate the biogeochemical processes that occur and their different impacts on the fate of these TE.To do so, we developed an original, experimental, column setup that mimics an aquifer with As-or Hg-iron-oxide rich areas versus iron depleted areas, enabling a better understanding of TE biogeochemistry in anoxic conditions. The following protocol gives step by step instructions for the column set-up either for As or Hg, as well as an example with As under iron and sulfate reducing conditions