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.

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.

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.

Arsenic scavenging by aluminum-substituted ferrihydrites in a circumneutral pH river impacted by acid mine drainage.

Ferrihydrite (Fh) is a nanocrystalline ferric oxyhydroxide involved in the retention of pollutants in natural systems and in water-treatment processes. The status and properties of major chemical impurities in natural Fh is however still scarcely documented. Here we investigated the structure of aluminum-rich Fh, and their role in arsenic scavenging in river-bed sediments from a circumneutral river (pH 6-7) impacted by an arsenic-rich acid mine drainage (AMD). Extended X-ray absorption fine structure (EXAFS) spectroscopy at the Fe K-edge shows that Fh is the predominant mineral phase forming after neutralization of the AMD, in association with minor amount of schwertmannite transported from the AMD. TEM-EDXS elemental mapping and SEM-EDXS analyses combined with EXAFS analysis indicates that Al(3+) substitutes for Fe(3+) ions into the Fh structure in the natural sediment samples, with local aluminum concentration within the 25-30 ± 10 mol %Al range. Synthetic aluminous Fh prepared in the present study are found to be less Al-substituted (14-20 ± 5 mol %Al). Finally, EXAFS analysis at the arsenic K-edge indicates that As(V) form similar inner-sphere surface complexes on the natural and synthetic Al-substituted Fh studied. Our results provide direct evidence for the scavenging of arsenic by natural Al-Fh, which emphasize the possible implication of such material for scavenging pollutants in natural or engineered systems.

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.

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.

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.

Child neurodevelopment in a Bolivian mining city.

This study evaluates the neurodevelopment of children living near contaminated mining industries during their first year of life. Participants from the city of Oruro (Bolivia) were prospectively recruited during pregnancy. Follow-up occurred between May 2007 and November 2009. Information about the socioeconomic status and medical history of the pregnant women were collected using questionnaires. Neurodevelopment was evaluated for 246 children using the Bayley Scales of Infant Development (BSID) at 10.5-12.5 months of age. Exposure to trace elements (Pb, As, Cd, Sb, Cs, Zn, Fe, Cu, Se, Rb, and Sr) during prenatal life was evaluated by testing maternal blood concentrations before delivery. Almost all measured levels were lower than the control limits. The blood lead concentration of pregnant women was low, considering the contaminated environmental context. The geometric mean was 1.76 µg/dL (95% CI: 1.68-1.84), a level comparable with those observed in non-contaminated areas. The only element found to be relatively elevated was antimony, with a geometric mean of 1.03 µg/dL (95% CI: 0.96-1.11). Our results suggest that women from this mining area were not highly exposed. The Bayley Scales of Infant Development (BSID) did not reveal mental or psychomotor abnormalities. Surprisingly, at the observed low levels, lead was positively associated with the children's BSID performance.

Surface properties and intracellular speciation revealed an original adaptive mechanism to arsenic in the acid mine drainage bio-indicator Euglena mutabilis.

Euglena mutabilis is a protist ubiquitously found in extreme environments such as acid mine drainages which are often rich in arsenic. The response of E. mutabilis to this metalloid was compared to that of Euglena gracilis, a protist not found in such environments. Membrane fatty acid composition, cell surface properties, arsenic accumulation kinetics, and intracellular arsenic speciation were determined. The results revealed a modification in fatty acid composition leading to an increased membrane fluidity in both Euglena species under sublethal arsenic concentrations exposure. This increased membrane fluidity correlated to an induced gliding motility observed in E. mutabilis in the presence of this metalloid but did not affect the flagellar dependent motility of E. gracilis. Moreover, when compared to E. gracilis, E. mutabilis showed highly hydrophobic cell surface properties and a higher tolerance to water-soluble arsenical compounds but not to hydrophobic ones. Finally, E. mutabilis showed a lower accumulation of total arsenic in the intracellular compartment and an absence of arsenic methylated species in contrast to E. gracilis. Taken together, our results revealed the existence of a specific arsenical response of E. mutabilis that may play a role in its hypertolerance to this toxic metalloid.

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.

Characterization of the active bacterial community involved in natural attenuation processes in arsenic-rich creek sediments.

Acid mine drainage of the Carnoulès mine (France) is characterized by acid waters containing high concentrations of arsenic and iron. In the first 30 m along the Reigous, a small creek draining the site, more than 38% of the dissolved arsenic was removed by co-precipitation with Fe(III), in agreement with previous studies, which suggest a role of microbial activities in the co-precipitation of As(III) and As(V) with Fe(III) and sulfate. To investigate how this particular ecosystem functions, the bacterial community was characterized in water and sediments by 16S rRNA encoding gene library analysis. Based on the results obtained using a metaproteomic approach on sediments combined with high-sensitivity HPLC-chip spectrometry, several GroEL orthologs expressed by the community were characterized, and the active members of the prokaryotic community inhabiting the creek sediments were identified. Many of these bacteria are ß-proteobacteria such as Gallionella and Thiomonas, but γ-proteobacteria such as Acidithiobacillus ferrooxidans and α-proteobacteria such as Acidiphilium, Actinobacteria, and Firmicutes were also detected.

Predominance of aqueous Tl(I) species in the river system downstream from the abandoned Carnoulès mine (Southern France).

Thallium concentration reached up to 534 µg L(-1) in the Reigous acid mine drainage downstream from the abandoned Pb-Zn Carnoulès mine (Southern France). It decreased to 5.44 µg L(-1) in the Amous River into which the Reigous creek flows. Tl(I) predominated (>98% of total dissolved Tl) over Tl(III), mainly in the form of Tl(+). Small amounts of Tl(III) evidenced in Reigous Creek might be in the form of aqueous TlCl(2)(+). The range of dissolved to particulate distribution coefficients log K(d) = 2.5 L kg(-1) to 4.6 L kg(-1) indicated low affinity of Tl for particles, mainly ferrihydrite, formed in the AMD-impacted watershed. The low retention of Tl(+) on ferrihydrite was demonstrated in sorption experiments, the best fit between experimental and modeled data being achieved for surface complexation constants log K(ads) = -2.67 for strong sites and log K(ads) = -3.76 for weak sites. This new set of constants allowed reasonable prediction of the concentrations of aqueous and particulate Tl resulting from the mixing of water from Reigous Creek and the Amous River water during laboratory experiments, together with those measured in the Amous River field study.

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.

Thermodesulfovibrio hydrogeniphilus sp. nov., a new thermophilic sulphate-reducing bacterium isolated from a Tunisian hot spring.

A new thermophilic sulphate-reducing bacterium (strain Hbr5T) was enriched and isolated from a terrestrial Tunisian hot spring. It was a non-spore-forming Gram-negative curved or vibrio-shaped bacterium. It appeared singly or in long chains and was actively motile by a polar flagellum. It possessed c-type cytochromes and desulfofuscidin. Growth occurred between 50 and 70 degrees C, with an optimum of 65 degrees C at pH 7.1. In the presence of sulphate as a terminal electron acceptor, this strain readily used H2 but formate only poorly. It could use sulphate, thiosulphate, sulphite or arsenate as electron acceptors. Its DNA G+C content was 36.1 mol%. Based on phenotypic, genomic, and phylogenetic characteristics, strain Hbr5T (=DSM 18151T, =JCM 13991T) is proposed to be assigned to a novel species of genus Thermodesulfovibrio, T. hydrogeniphilus sp. nov.

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.

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.

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.