Study of microbial communities (bacteria, archaea and eukaryota) and their spatiotemporal variations in Carnoulès mine highly contaminated in arsenic

L'ancienne mine de plomb et de zinc de Carnoulès (Gard, France) a généré 1.5 Mt de déchets d'où émerge un drainage de mine aux eaux acides alimentant un ruisseau, le Reigous. Ce site fournit un exceptionnel exemple d'adaptation à un environnement extrême en raison des eaux acides (pH~3) et de très fortes concentrations en métaux et métalloïdes, particulièrement en As. Dans les 30 premiers mètres du ruisseau, l'activité bactérienne conduit à un phénomène de remédiation naturelle avec la co-précipitation de 20 à 60% de l'As dissous avec du fer. Les bactéries présentes dans les sédiments du ruisseau ont dans un premier temps été décrites par clonage/séquençage du gène de l'ARNr 16S, puis les membres actifs des communautés bactériennes ont été révélés par une approche de métaprotéomique. L'étude des Archaea au sein des sédiments a révélé la présence de groupes impliqués dans la méthanogénèse ou dans l'oxydation de l'ammoniac qui pourraient participer au cycle du carbone ou de l'azote. Les eucaryotes ont été caractérisés pour la première fois sur ce site par pyroséquençage, mettant en évidence une forte proportion de champignons (60%). Enfin, l'étude des variations spatiotemporelles des populations bactériennes dans les eaux a conduit à l'identification de 6801 OTUs dont des phyla encore jamais identifiés sur ce site. La concentration en arsenic, la température et le potentiel redox semblent jouer un rôle dans la structuration de ces communautés. Ce travail de thèse a ainsi contribué à une meilleure connaissance des microorganismes présents (Bactéries, Archaea, Eucaryotes) et de leurs dynamiques spatiotemporelles en relation avec les paramètres physicochimiques du milieu.Acidic mining drainage generated at Carnoulès, a former Pb-Zn mine (Gard, France) coincides with the spring of the Reigous Creek. This site provides an exceptional example of adaptation to extreme environments due to its acidic water (pH~3) and very high concentrations of metals and metalloids, particularly arsenic. During the first 30 m of downflow in Reigous Creek, natural remediation occurred, with co-precipitation of 20 to 60% of the dissolved arsenic with iron, mediated by bacteria. Bacterial communities inhabiting the creek sediments were first described by cloning/sequencing of the 16S rRNA genes and the active members were identified by a metaproteomic approach. A survey of the archaeal community in the sediment highlighted the presence of sequences phylogenetically related to methanogenic Archaea and to ammonia oxidizers, which could be involved in carbon and nitrogen biochemical cycling. The Eukaryotic communities were studied for the first time at this site by pyrosequencing, revealing that around 60% of the sequences belonged to Fungi. Finally, the study of the spatiotemporal variations of the water bacterial communities allowed the identification of 6801 OTUs including sequences of taxa never detected before. The environmental variables significantly correlated with bacterial community dynamics appear to be arsenic concentration, temperature and Eh. This PhD work has contributed to a better understanding of the spatiotemporal dynamics of microorganisms (Bacteria, Archaea, Eukaryotes) in relation with the physicochemical parameters of their environment

Determination of potential denitrification in a range of tropical topsoils using near infrared reflectance spectroscopy (NIRS)

Microbial denitrification plays a central role in nitrous oxide (N2O)-emitting processes, which are involved in ecosystem services such as crop production and climate regulation. Field characterization of N2O-emitting processes being time-consuming due to great variability, laboratory determination of potential denitrification (upon incubation) is often used as a valuable test. Near infrared reflectance spectroscopy (NIRS) is a time- and cost-effective approach that has been reported to allow accurate determination of several soil properties. The objective of the present study was to assess the interest of NIRS for determining potential denitrification over a set of 460 topsoils sampled under crops, tree plantations, savanna or rainforest, originating from Madagascar, Congo-Brazzaville, Brazil, and French Guiana. Prediction of potential denitrification using NIRS was satisfying over the total set, especially with LOCAL calibration, which builds a model for each sample separately using its spectral neighbours in the calibration subset (R-2=0.79 for validation). For the other sets, either textural or geographical, global calibration only was performed, involving for each set a unique prediction model built with all calibration samples. The accuracy of NIRS determination depended on the sample set, decreasing in the following order: Malagasy clayey set > total set approximate to. Brazilian sandy loam set > coarse-textured set (Congo-Brazil) approximate to Guianese sandy clay loam set approximate to Congolese sandy set > non-clayey set (Congo-Brazil-Guiana), with cross-validation R-2 ranging from 0.88 to 0.44 (external validation was not carried out for small-sized sets). Thus NIRS prediction was more accurate over the clayey homogeneous set than over the non-clayey heterogeneous set. As a result of global calibration, potential denitrification was expressed as a linear combination of absorbance at every wavelength. Wavelengths that contributed most to NIRS prediction of soil potential denitrification corresponded to wavelengths that literature has assigned to organic nitrogenous compounds, amide-containing ones especially, and to carbonaceous compounds such as cellulose or including CH3 or CH2 groups. This related to the importance of amides in soil organic nitrogen and microbial biomass, and to the dependence of denitrification on soil organic matter. In short, NIRS is a time- and cost-effective approach that proved relevant for determining soil potential denitrification with acceptable accuracy, especially for clayey samples or when LOCAL calibration was performed