Towards an understanding of the factors controlling bacterial diversity and activity in semi-passive Fe- and As-oxidizing bioreactors treating arsenic-rich acid mine drainage.

Semi-passive bioreactors based on iron and arsenic oxidation and coprecipitation are promising for the treatment of As-rich acid mine drainages. However, their performance in the field remains variable and unpredictable. Two bioreactors filled with distinct biomass carriers (plastic or a mix of wood and pozzolana) were monitored during 1 year. We characterized the dynamic of the bacterial communities in these bioreactors, and explored the influence of environmental and operational drivers on their diversity and activity. Bacterial diversity was analyzed by 16S rRNA gene metabarcoding. The aioA genes and transcripts were quantified by qPCR and RT-qPCR. Bacterial communities were dominated by several iron-oxidizing genera. Shifts in the communities were attributed to operational and physiochemical parameters including the nature of the biomass carrier, the water pH, temperature, arsenic, and iron concentrations. The bioreactor filled with wood and pozzolana showed a better resilience to disturbances, related to a higher bacterial alpha diversity. We evidenced for the first time aioA expression in a treatment system, associated with the presence of active Thiomonas spp. This confirmed the contribution of biological arsenite oxidation to arsenic removal. The resilience and the functional redundancy of the communities developed in the bioreactors conferred robustness and stability to the treatment systems.

Bio-precipitation of arsenic and antimony in a sulfate-reducing bioreactor treating real acid mine drainage water.

Arsenic (As) and antimony (Sb) from mining sites can seep into aquatic ecosystems by acid mine drainage (AMD). Here, the possibility of concomitantly removing As and Sb from acidic waters by precipitation of sulfides induced by sulfate-reducing bacteria (SRB) was investigated in a fixed-bed column bioreactor. The real AMD water used to feed the bioreactor contained nearly 1 mM As, while the Sb concentrations were increased (0.008 ± 0.006 to 1.01 ± 0.07 mM) to obtain an Sb/As molar ratio = 1. Results showed that the addition of Sb did not affect the efficiency of As bio-precipitation. Sb was removed efficiently (up to 97.9% removal) between the inlet and outlet of the bioreactor, together with As (up to 99.3% removal) in all conditions. Sb was generally removed as it entered the bioreactor. Appreciable sulfate reduction occurred in the bioreactor, which could have been linked to the stable presence of a major SRB operational taxonomic unit affiliated with the Desulfosporosinus genus. The bacterial community included polymer degraders, fermenters, and acetate degraders. Results suggested that sulfate reduction could be a suitable bioremediation process for the simultaneous removal of Sb and As from AMD.

Impact of past mining activities on water quality in a karst area in the Cévennes region, Southern France.

Sampling and analysis of groundwater and surface water were conducted to assess the potential impacts of abandoned mines on water quality in a karst area in Southern France. The results of multivariate statistical analysis and geochemical mapping revealed that water quality is affected by contaminated drainage from abandoned mine sites. Acid mine drainage with very high concentrations of Fe, Mn, Al, Pb and Zn was identified in a few samples collected from mine openings and near waste dumps. In general, neutral drainage with elevated concentrations of Fe, Mn, Zn, As, Ni and Cd was observed due to buffering by carbonate dissolution. The contamination is spatially limited around abandoned mine sites, suggesting that metal(oid)s are sequestered in secondary phases that form under near-neutral and oxidizing conditions. However, the analysis of seasonal variations in trace metal concentrations showed that the transport of metal contaminants in water is highly variable according to hydrological conditions. During low flow conditions, trace metals are likely to be rapidly sequestered in Fe-oxyhydroxides and carbonate minerals in the karst aquifer and the river sediments, while low or no surface runoff in intermittent rivers limits the transport of contaminants in the environment. On the other hand, significant amounts of metal(loid)s can be transported under high flow conditions, primarily in dissolved form. Dissolved metal(loid) concentrations in groundwater remained elevated despite dilution by uncontaminated water, likely as a result of the increased leaching of mine wastes and the flushing of contaminated waters from mine workings. This work shows that groundwater is the main source of contamination to the environment and highlights the need to better understand the fate of trace metals in karst water systems.

Development and implementation of a multi-criteria aggregation operator to estimate the contributions of the natural geochemical background and anthropogenic inputs in groundwater in former mining regions: An application to arsenic and antimony in the Gardon river watershed (southern France).

Establishing the contribution of natural enrichment of a substance and anthropogenic inputs has become a major issue for the management of groundwater systems. The issue is made more complex when the geology of the concerned territories is heterogeneous, at variable geographical scales, at a site that has experienced mining activity that has left behind mining remains. Several studies have tried to answer this problem using different approaches: statistical, geostatistical, geological, and geochemical. The limits of these studies are seen through the incomplete integration of geological and geomorphological parameters in the results. The aim of the present research is, therefore, to look deeper into an approach to estimate the respective contributions of the natural geochemical background and anthropogenic inputs, by simultaneously considering the heterogeneity of the geology, the variability of the spatial scale, and the combination of geological, geomorphological and statistical factors. A multi-criteria aggregation operator was thus developed and implemented on underground water bodies delimited by the Gardon watershed in the Cevennes (a region with former mining activities - France), in order to produce quantitative and qualitative maps for discriminating between the natural geochemical background and anthropogenic inputs. 176 geochemical observation points on groundwater quality were collected by sampling and through the acquisition of public data on water sources (ADES database), wells and boreholes, to reconstruct the spatial distribution of arsenic and antimony in the study site. An aggregation operator was developed, which enabled the determination of the formulae to calculate the natural geochemical background and hence deduce the anthropogenic contributions. Cartography of the quantitative and qualitative aspects of the geochemical concentrations that have been impacted by anthropogenic activity made it possible to determine remarkable focal points located on the Cevennes fault and other specific points of geochemical interest.

Modeling of microbial kinetics and mass transfer in bioreactors simulating the natural attenuation of arsenic and iron in acid mine drainage.

Natural attenuation in acid mine drainage (AMD) due to biological iron and arsenic oxidation offers a promising strategy to treat As-rich AMD in passive bioreactors. A reactive transport model is developed in order to identify the main controlling factors. It simulates batch and flow-through experiments that reproduce natural attenuation in a high-As AMD. The 2-D model couples second-order microbial kinetics (Fe- and As- oxidation) and geochemical reactions to hydrodynamic transport. Oxidation only occurrs in the biofilm with an oxygen transfer from the air through the water column. The model correctly simulates the Fe(II)-Fe(III) and As(III)-As(V) concentrations in the outlet waters and the precipitates, over hydraulic retention times from 30 min to 800 min. It confirms that the natural attenuation at 20 °C is driven by the fast Fe(II) oxidation and slow As(III) oxidation that favors arsenite trapping by schwertmannite over amorphous ferric arsenate (AFA) formation. The localization of iron oxidation in the biofilm limits the attenuation of arsenic and iron as the water column height increases. The change in the composition of the bacterial iron-oxidizer community of the biofilm at the lowest pH boundary seems to control the Fe(II) oxidation kinetic rate besides the bacterial concentration.

Contrasting arsenic biogeochemical cycling in two Moroccan alkaline pit lakes.

Pit lakes resulting from the flooding of abandoned mines represent a valuable freshwater reserve. However, water contamination by toxic elements, including arsenic, compromises their use for freshwater supply. For a better management of these reserves, our aim was to gain insight into arsenic cycling in two Moroccan alkaline pit lakes. We first showed that dimethylarsenic dominated in stratified lake ZA whereas in lake ZL1, As(V) was prevailing. Because microbially mediated processes largely contribute to arsenic cycling, the diversity of arsenic-methylating and -oxidizing bacteria was determined through the sequencing of arsM and aioA genes. Diverse arsM-carrying bacteria were thriving in ZA while a low diversity of aioA genes was detected in ZL1. We also determined the structure of the total bacterial communities by fingerprinting (ARISA). Contrasting arsenic speciation and bacterial communities in the two lakes were associated with differences of conductivity, Total Organic Carbon and temperature. In ZA, dissolved oxygen and redox potential were the main factors driving the total bacterial community structure and the ArsM diversity. In ZL1, stable bacterial communities were associated with limited water physico-chemistry variations. Our study provides new insights into the biogeochemical behavior of arsenic and the role of arsenic transforming bacteria in alkaline pit lakes.

Hydraulic retention time affects bacterial community structure in an As-rich acid mine drainage (AMD) biotreatment process.

Arsenic removal consecutive to biological iron oxidation and precipitation is an effective process for treating As-rich acid mine drainage (AMD). We studied the effect of hydraulic retention time (HRT)-from 74 to 456 min-in a bench-scale bioreactor exploiting such process. The treatment efficiency was monitored during 19 days, and the final mineralogy and bacterial communities of the biogenic precipitates were characterized by X-ray absorption spectroscopy and high-throughput 16S rRNA gene sequencing. The percentage of Fe(II) oxidation (10-47%) and As removal (19-37%) increased with increasing HRT. Arsenic was trapped in the biogenic precipitates as As(III)-bearing schwertmannite and amorphous ferric arsenate, with a decrease of As/Fe ratio with increasing HRT. The bacterial community in the biogenic precipitate was dominated by Fe-oxidizing bacteria whatever the HRT. The proportion of Gallionella and Ferrovum genera shifted from respectively 65 and 12% at low HRT to 23 and 51% at high HRT, in relation with physicochemical changes in the treated water. aioA genes and Thiomonas genus were detected at all HRT although As(III) oxidation was not evidenced. To our knowledge, this is the first evidence of the role of HRT as a driver of bacterial community structure in bioreactors exploiting microbial Fe(II) oxidation for AMD treatment.

Temperature and nutrients as drivers of microbially mediated arsenic oxidation and removal from acid mine drainage.

Microbial oxidation of iron (Fe) and arsenic (As) followed by their co-precipitation leads to the natural attenuation of these elements in As-rich acid mine drainage (AMD). The parameters driving the activity and diversity of bacterial communities responsible for this mitigation remain poorly understood. We conducted batch experiments to investigate the effect of temperature (20 vs 35 °C) and nutrient supply on the rate of Fe and As oxidation and precipitation, the bacterial diversity (high-throughput sequencing of 16S rRNA gene), and the As oxidation potential (quantification of aioA gene) in AMD from the Carnoulès mine (France). In batch incubated at 20 °C, the dominance of iron-oxidizing bacteria related to Gallionella spp. was associated with almost complete iron oxidation (98%). However, negligible As oxidation led to the formation of As(III)-rich precipitates. Incubation at 35 °C and nutrient supply both stimulated As oxidation (71-75%), linked to a higher abundance of aioA gene and the dominance of As-oxidizing bacteria related to Thiomonas spp. As a consequence, As(V)-rich precipitates (70-98% of total As) were produced. Our results highlight strong links between indigenous bacterial community composition and iron and arsenic removal efficiency within AMD and provide new insights for the future development of a biological treatment of As-rich AMD.

Dynamics of Bacterial Communities Mediating the Treatment of an As-Rich Acid Mine Drainage in a Field Pilot.

Passive treatment based on iron biological oxidation is a promising strategy for Arsenic (As)-rich acid mine drainage (AMD) remediation. In the present study, we characterized by 16S rRNA metabarcoding the bacterial diversity in a field-pilot bioreactor treating extremely As-rich AMD in situ, over a 6 months monitoring period. Inside the bioreactor, the bacterial communities responsible for iron and arsenic removal formed a biofilm ("biogenic precipitate") whose composition varied in time and space. These communities evolved from a structure at first similar to the one of the feed water used as an inoculum to a structure quite similar to the natural biofilm developing in situ in the AMD. Over the monitoring period, iron-oxidizing bacteria always largely dominated the biogenic precipitate, with distinct populations (Gallionella, Ferrovum, Leptospirillum, Acidithiobacillus, Ferritrophicum), whose relative proportions extensively varied among time and space. A spatial structuring was observed inside the trays (arranged in series) composing the bioreactor. This spatial dynamic could be linked to the variation of the physico-chemistry of the AMD water between the raw water entering and the treated water exiting the pilot. According to redundancy analysis (RDA), the following parameters exerted a control on the bacterial communities potentially involved in the water treatment process: dissolved oxygen, temperature, pH, dissolved sulfates, arsenic and Fe(II) concentrations and redox potential. Appreciable arsenite oxidation occurring in the bioreactor could be linked to the stable presence of two distinct monophylogenetic groups of Thiomonas related bacteria. The ubiquity and the physiological diversity of the bacteria identified, as well as the presence of bacteria of biotechnological relevance, suggested that this treatment system could be applied to the treatment of other AMD.

The environmental legacy of historic Pb-Zn-Ag-Au mining in river basins of the southern edge of the Massif Central (France).

The main rivers (Aude, Orb, Herault) that discharge into the Gulf of Lions and the west bank tributaries of the Rhone River including the Gardon have former non-ferrous metal mines in their upper drainage basin. Using unpublished data and data from the literature, this study provides an integrated overview of the contamination of water and sediment along the continent-sea continuum and of its impacts on the biota and on human health. In the upper part of these basins, water and stream sediments are enriched in metal(-loids) compared to median European concentrations. Arsenic is the main contaminant in the rivers Aude and Gardon d'Anduze, Sb in the Orb and Gardon d'Alès, and Tl in the Herault river. A rapid reduction in dissolved and particulate concentrations was systematically observed along the river due to dilution and precipitation. The high concentrations of metal(-loid)s observed suggest that the former mining activity still represents a potential threat for the environment, but the lack of high temporal resolution monitoring, especially during Mediterranean floods, prevents accurate assessment of metal fluxes from these rivers to the Mediterranean Sea. Studies dedicated to the impacts on human health are too rare, given that studies have shown a higher rate of arsenic-specific cancer near Salsigne mine in the Aude River basin and cases of saturnism in children in the upper Herault River basin. These studies underline the need to take environmental health issues into consideration not only in these watersheds but around the entire Mediterranean basin, which harbors numerous metalliferous ores that have been mined for millennia.

Thallium release from acid mine drainages: Speciation in river and tap water from Valdicastello mining district (northwest Tuscany).

In this work we present an advantageous method for the simultaneous separation and detection of Tl(I) and Tl(III) species through ion chromatography coupled with on-line inductively coupled plasma - mass spectrometry. Chromatographic separation between Tl(III) and Tl(I) was achieved in less than two minutes. The method was validated by recovery experiments on real samples, and by comparing the sum of the concentrations of individual Tl species with total thallium values obtained from continuous flow ICP-MS. The experimental procedure offers an accurate, sensitive and interference-free method for Tl speciation at trace levels in environmental samples. This allowed us to investigate the Tl speciation in acid mine drainages (AMD), surface waters and springs in a mining catchment in Valdicastello Carducci (Tuscany, Italy), where severe Tl contamination ad been evidenced previously. This study shows for the first time that Tl(III), in addition to Tl(I), is present in considerable amounts in water samples affected by acid mining outflow, raising the question of the origin of this thermodynamically unstable species.

Complete removal of arsenic and zinc from a heavily contaminated acid mine drainage via an indigenous SRB consortium.

Acid mine drainages (AMD) are major sources of pollution to the environment. Passive bio-remediation technologies involving sulfate-reducing bacteria (SRB) are promising for treating arsenic contaminated waters. However, mechanisms of biogenic As-sulfide formation need to be better understood to decontaminate AMDs in acidic conditions. Here, we show that a high-As AMD effluent can be decontaminated by an indigenous SRB consortium. AMD water from the Carnoulès mine (Gard, France) was incubated with the consortium under anoxic conditions and As, Zn and Fe concentrations, pH and microbial activity were monitored during 94days. Precipitated solids were analyzed using electron microscopy (SEM/TEM-EDXS), and Extended X-Ray Absorption Fine Structure (EXAFS) spectroscopy at the As K-edge. Total removal of arsenic and zinc from solution (1.06 and 0.23mmol/L, respectively) was observed in two of the triplicates. While Zn precipitated as ZnS nanoparticles, As precipitated as amorphous orpiment (am-AsIII2S3) (33-73%), and realgar (AsIIS) (0-34%), the latter phase exhibiting a particular nanowire morphology. A minor fraction of As is also found as thiol-bound AsIII (14-23%). We propose that the formation of the AsIIS nanowires results from AsIII2S3 reduction by biogenic H2S, enhancing the efficiency of As removal. The present description of As immobilization may help to set the basis for bioremediation strategies using SRB.

Spatio-Temporal Detection of the Thiomonas Population and the Thiomonas Arsenite Oxidase Involved in Natural Arsenite Attenuation Processes in the Carnoulès Acid Mine Drainage.

The acid mine drainage (AMD) impacted creek of the Carnoulès mine (Southern France) is characterized by acid waters with a high heavy metal content. The microbial community inhabiting this AMD was extensively studied using isolation, metagenomic and metaproteomic methods, and the results showed that a natural arsenic (and iron) attenuation process involving the arsenite oxidase activity of several Thiomonas strains occurs at this site. A sensitive quantitative Selected Reaction Monitoring (SRM)-based proteomic approach was developed for detecting and quantifying the two subunits of the arsenite oxidase and RpoA of two different Thiomonas groups. Using this approach combined with FISH and pyrosequencing-based 16S rRNA gene sequence analysis, it was established here for the first time that these Thiomonas strains are ubiquitously present in minor proportions in this AMD and that they express the key enzymes involved in natural remediation processes at various locations and time points. In addition to these findings, this study also confirms that targeted proteomics applied at the community level can be used to detect weakly abundant proteins in situ.

Arsenite response in Coccomyxa sp. Carn explored by transcriptomic and non-targeted metabolomic approaches.

Arsenic is a toxic metalloid known to generate an important oxidative stress in cells. In the present study, we focused our attention on an alga related to the genus Coccomyxa, exhibiting an extraordinary capacity to resist high concentrations of arsenite and arsenate. The integrated analysis of high-throughput transcriptomic data and non-targeted metabolomic approaches highlighted multiple levels of protection against arsenite. Indeed, Coccomyxa sp. Carn induced a set of transporters potentially preventing the accumulation of this metalloid in the cells and presented a distinct arsenic metabolism in comparison to another species more sensitive to that compound, i.e. Euglena gracilis, especially in regard to arsenic methylation. Interestingly, Coccomyxa sp. Carn was characterized by a remarkable accumulation of the strong antioxidant glutathione (GSH). Such observation could explain the apparent low oxidative stress in the intracellular compartment, as suggested by the transcriptomic analysis. In particular, the high amount of GSH in the cell could play an important role for the tolerance to arsenate, as suggested by its partial oxidation into oxidized glutathione in presence of this metalloid. Our results therefore reveal that this alga has acquired multiple and original defence mechanisms allowing the colonization of extreme ecosystems such as acid mine drainages.

Diversity and Distribution of Arsenic-Related Genes Along a Pollution Gradient in a River Affected by Acid Mine Drainage.

Some microorganisms have the capacity to interact with arsenic through resistance or metabolic processes. Their activities contribute to the fate of arsenic in contaminated ecosystems. To investigate the genetic potential involved in these interactions in a zone of confluence between a pristine river and an arsenic-rich acid mine drainage, we explored the diversity of marker genes for arsenic resistance (arsB, acr3.1, acr3.2), methylation (arsM), and respiration (arrA) in waters characterized by contrasted concentrations of metallic elements (including arsenic) and pH. While arsB-carrying bacteria were representative of pristine waters, Acr3 proteins may confer to generalist bacteria the capacity to cope with an increase of contamination. arsM showed an unexpected wide distribution, suggesting biomethylation may impact arsenic fate in contaminated aquatic ecosystems. arrA gene survey suggested that only specialist microorganisms (adapted to moderately or extremely contaminated environments) have the capacity to respire arsenate. Their distribution, modulated by water chemistry, attested the specialist nature of the arsenate respirers. This is the first report of the impact of an acid mine drainage on the diversity and distribution of arsenic (As)-related genes in river waters. The fate of arsenic in this ecosystem is probably under the influence of the abundance and activity of specific microbial populations involved in different As biotransformations.

Toxic trace elements in maternal and cord blood and social determinants in a Bolivian mining city.

This study assessed lead, arsenic, and antimony in maternal and cord blood, and associations between maternal concentrations and social determinants in the Bolivian mining city of Oruro using the baseline assessment of the ToxBol/Mine-Niño birth cohort. We recruited 467 pregnant women, collecting venous blood and sociodemographic information as well as placental cord blood at birth. Metallic/semimetallic trace elements were measured using inductively coupled plasma mass spectrometry. Lead medians in maternal and cord blood were significantly correlated (Spearman coefficient = 0.59; p < 0.001; 19.35 and 13.50 µg/L, respectively). Arsenic concentrations were above detection limit (3.30 µg/L) in 17.9% of maternal and 34.6% of cord blood samples. They were not associated (Fischer's p = 0.72). Antimony medians in maternal and cord blood were weakly correlated (Spearman coefficient = 0.15; p < 0.03; 9.00 and 8.62 µg/L, respectively). Higher concentrations of toxic elements in maternal blood were associated with maternal smoking, low educational level, and partner involved in mining.

Antimony isotopic composition in river waters affected by ancient mining activity.

In this study, antimony (Sb) isotopic composition was determined in natural water samples collected along two hydrosystems impacted by historical mining activities: the upper Orb River and the Gardon River watershed (SE, France). Antimony isotope ratio was measured by HG-MC-ICP-MS (Hydride Generation Multi-Collector Inductively Coupled Plasma Mass Spectrometer) after a preconcentration and purification step using a new thiol-cellulose powder (TCP) procedure. The external reproducibility obtained for δ(123)Sb measurements of our in-house Sb isotopic standard solution and a certified reference freshwater was 0.06‰ (2σ). Significant isotopic variations were evident in surface waters from the upper Orb River (-0.06‰≤δ(123)Sb≤+0.11‰) and from the Gardon River watershed (+0.27‰≤δ(123)Sb≤+0.83‰). In particular, streams that drained different former mining sites exploited for Sb or Pb-Zn exhibited contrasted Sb isotopic signature, that may be related to various biogeochemical processes occurring during Sb transfer from rocks, mine wastes and sediments to the water compartment. Nevertheless, Sb isotopic composition appeared to be stable along the Gardon River, which might be attributed to the conservative transport of Sb at distance from mine-impacted streams, due to the relative mobile behavior of Sb(V) in natural oxic waters. This study suggests that Sb isotopic composition could be a useful tool to track pollution sources and/or biogeochemical processes in hydrologic systems.

Thiomonas sp. CB2 is able to degrade urea and promote toxic metal precipitation in acid mine drainage waters supplemented with urea.

The acid mine drainage (AMD) in Carnoulès (France) is characterized by the presence of toxic metals such as arsenic. Several bacterial strains belonging to the Thiomonas genus, which were isolated from this AMD, are able to withstand these conditions. Their genomes carry several genomic islands (GEIs), which are known to be potentially advantageous in some particular ecological niches. This study focused on the role of the "urea island" present in the Thiomonas CB2 strain, which carry the genes involved in urea degradation processes. First, genomic comparisons showed that the genome of Thiomonas sp. CB2, which is able to degrade urea, contains a urea genomic island which is incomplete in the genome of other strains showing no urease activity. The urease activity of Thiomonas sp. CB2 enabled this bacterium to maintain a neutral pH in cell cultures in vitro and prevented the occurrence of cell death during the growth of the bacterium in a chemically defined medium. In AMD water supplemented with urea, the degradation of urea promotes iron, aluminum and arsenic precipitation. Our data show that ureC was expressed in situ, which suggests that the ability to degrade urea may be expressed in some Thiomonas strains in AMD, and that this urease activity may contribute to their survival in contaminated environments.

Diversity and spatiotemporal dynamics of bacterial communities: physicochemical and other drivers along an acid mine drainage.

Deciphering the biotic and abiotic factors that control microbial community structure over time and along an environmental gradient is a pivotal question in microbial ecology. Carnoulès mine (France), which is characterized by acid waters and very high concentrations of arsenic, iron, and sulfate, provides an excellent opportunity to study these factors along the pollution gradient of Reigous Creek. To this end, biodiversity and spatiotemporal distribution of bacterial communities were characterized using T-RFLP fingerprinting and high-throughput sequencing. Patterns of spatial and temporal variations in bacterial community composition linked to changes in the physicochemical conditions suggested that species-sorting processes were at work in the acid mine drainage. Arsenic, temperature, and sulfate appeared to be the most important factors that drove the composition of bacterial communities along this continuum. Time series investigation along the pollution gradient also highlighted habitat specialization for some major members of the community (Acidithiobacillus and Thiomonas), dispersal for Acidithiobacillus, and evidence of extinction/re-thriving processes for Gallionella. Finally, pyrosequencing revealed a broader phylogenetic range of taxa than previous clone library-based diversity. Overall, our findings suggest that in addition to environmental filtering processes, additional forces (dispersal, birth/death events) could operate in AMD community.

Persisting impact of historical mining activity to metal (Pb, Zn, Cd, Tl, Hg) and metalloid (As, Sb) enrichment in sediments of the Gardon River, Southern France.

In this study, we assessed past and present influence of ancient mining activity on metal(loid) enrichment in sediments of a former mining watershed (Gardon River, SE France), that is now industrialized and urbanized. A sedimentary archive and current sediments were characterized combining geochemical analyses, zinc isotopic analyses and sequential extractions. The archive was used to establish local geochemical background and recorded (i) increasing enrichment factors (EFs) for Pb, Zn, Cd, Tl, Hg, As and Sb throughout the industrial era, (ii) a contamination peak in 1976 attributed to a tailings dam failure, and (iii) current levels in 2002 and 2011 similar to those of 1969, except for Sb and Hg, reflecting a persisting contamination pattern. Inter-element relationships and spatial distribution of EF values of current sediments throughout the watershed suggested that both ancient and current contamination had a common origin for Pb, Zn, Cd, Tl and As related to the exploitation of Pb/Zn mineralization while old Sb mines and coal extraction area were the main sources for Sb and Hg respectively. This prevailing mining origin was reflected for Zn by a relatively uniform isotopic composition at δ(66)Zn=0.23 ± 0.03‰, although slight decrease from 0.23‰ to 0.18‰ was recorded from upstream to downstream sites along the river course in relation with the contribution of the lighter δ(66)Zn signature (~0.08‰) of acid mine drainage impacted tributaries. Results from sequential extractions revealed that the potential mobility of the studied metal(loid)s varied in the order Sb