Use of a continuous-flow bioreactor to evaluate nitrate reduction rate of Halomonas desiderata in cementitious environment relevant to nuclear waste deep repository

The redox level of repositories can influence the mobility of the waste components stored in them (i.e. radionuclides) and thus the related safety assessments. Microbial activity is known to impact the control of redox reactions, the mechanisms and kinetics of which must be evaluated. This study investigates the denitrification rates of a model bacterium Halomonas desiderata (Hd) in cementitious environment with alkaline and anoxic conditions comparable to those found in radioactive waste repository cells. The growth and the total oxidized nitrogen (TON) reduction rates of Hd was determined in a continuous bioreactor with several feeding solutions with or without solid cement paste. Temporary nitrite accumulation and reduced denitrification rates are correlated with diminished bacterial growth. When the system was fed by optimal culture medium supplemented with acetate and nitrate, the TON reduction rates varied between 0.082 mM TON/h and 0.063 mM TON/h, depending on whether solid cement paste was present in the reactor or not. When the culture medium was replaced with pure cement leachate, the reaction rates increased to 0.137 mM TON/h with solid cement paste and dropped to 0.023 mM TON/h without. In these conditions at pH 10, solid cement paste had no negative influence on Hd activity

STED nanoscopy : A novel way to image the pore space of geological materials

STED nanoscopy (Stimulated Emission Depletion). which can resolve details far below the diffraction barrier has been applied hitherto preferentially to life sciences. The method is however also ideal for the investigation of geological matrices containing transparent minerals, an application tested here, to our knowledge, for the first time. The measurements on altered granitic rock and sedimentary clay rock, both containing very fine-grained phases, were conducted successfully. The STED fluorophore was dissolved in C-14-labelled methylmethacrylate (C-14-MMA) monomer which was polymerised within the rock matrix, thereby labelling the pore space in the geomaterials. Double labelling provided by the C-14-labelled MMA enables autoradiography and scanning electron microscopy (SEM), providing necessary complementary information for characterisation and quantification of porosity distributions and mineral and structure identification. Promising perspectives for further investigations of geological matrices by using different fluorophores and the optimisation of measuring procedures or even higher resolution are discussed. The combination of these different methods enlarges the observation scale of porosity from nanometre to centimetre scale.Peer reviewe

Dielectric relaxation behavior of Callovo-Oxfordian clay rock: A hydraulic-mechanical-electromagnetic coupling approach

Water content is a key parameter to monitor in nuclear waste repositories such as the planed underground repository in Bure, France, in the Callovo-Oxfordian (COx) clay formation. High-frequency electromagnetic (HF-EM) measurement techniques, i.e., time or frequency domain reflectometry, offer useful tools for quantitative estimation of water content in porous media. However, despite the efficiency of HF-EM methods, the relationship between water content and dielectric material properties needs to be characterized. Moreover, the high amount of swelling clay in the COx clay leads to dielectric relaxation effects which induce strong dispersion coupled with high absorption of EM waves. Against this background, the dielectric relaxation behavior of the clay rock was studied at frequencies from 1 MHz to 10 GHz with network analyzer technique in combination with coaxial transmission line cells. For this purpose, undisturbed and disturbed clay rock samples were conditioned to achieve a water saturation range from 0.16 to nearly saturation. The relaxation behavior was quantified based on a generalized fractional relaxation model under consideration of an apparent direct current conductivity assuming three relaxation processes: a high-frequency water process and two interface processes which are related to interactions between the aqueous pore solution and mineral particles (adsorbed/hydrated water relaxation, counter ion relaxation and Maxwell-Wagner effects). The frequency-dependent HF-EM properties were further modeled based on a novel hydraulic-mechanical-electromagnetic coupling approach developed for soils. The results show the potential of HF-EM techniques for quantitative monitoring of the hydraulic state in underground repositories in clay formations

Nitrate and nitrite bacterial reduction at alkaline pH and high nitrate concentrations, comparison of acetate versus dihydrogen as electron donors

This study assesses bacterial denitrification at alkaline pH, up to 12, and high nitrate concentration, up to 400 mM. Two types of electron donors organic (acetate) and inorganic (dihydrogen) were compared. With both types of electron donors, nitrite reduction was the key step, likely to increase the pH and lead to nitrite accumulation. Firstly, an acclimation process was used: nitrate was progressively increased in three cultures set at pH 9, 10, or 11. This method allowed to observe for the first time nitrate reduction up to pH 10 and 100 mM nitrate with dihydrogen, or up to pH 10 and 400 mM nitrate with acetate. Nitrate reduction kinetics were faster in the presence of acetate. To investigate further the impact of the type of electron donor, a transition from acetate to dihydrogen was tested, and the pH evolution was modelled. Denitrification with dihydrogen strongly increases the pH while with acetate the pH evolution depends on the initial pH. The main difference is the production of acidifying CO2 during the acetate oxidation. Finally, the use of long duration cultures with a highly alkaline pH allowed a nitrate reduction up to pH 11.5 with acetate. However, no reduction was possible in hydrogenotrophy as it would have increased the pH further. Instead, bacteria used organic matter from inoculum to reduce nitrate at pH 11.5. Therefore, considering bacterial denitrification in a context of alkaline pH and high nitrate concentration an organic electron donor such as acetate is advantageous

Dielectric relaxation behavior of Callovo-Oxfordian clay rock: A hydraulic-mechanical-electromagnetic coupling approach

Water content is a key parameter to monitor in nuclear waste repositories such as the planed underground repository in Bure, France, in the Callovo-Oxfordian (COx) clay formation. High-frequency electromagnetic (HF-EM) measurement techniques, i.e., time or frequency domain reflectometry, offer useful tools for quantitative estimation of water content in porous media. However, despite the efficiency of HF-EM methods, the relationship between water content and dielectric material properties needs to be characterized. Moreover, the high amount of swelling clay in the COx clay leads to dielectric relaxation effects which induce strong dispersion coupled with high absorption of EM waves. Against this background, the dielectric relaxation behavior of the clay rock was studied at frequencies from 1 MHz to 10 GHz with network analyzer technique in combination with coaxial transmission line cells. For this purpose, undisturbed and disturbed clay rock samples were conditioned to achieve a water saturation range from 0.16 to nearly saturation. The relaxation behavior was quantified based on a generalized fractional relaxation model under consideration of an apparent direct current conductivity assuming three relaxation processes: a high-frequency water process and two interface processes which are related to interactions between the aqueous pore solution and mineral particles (adsorbed/hydrated water relaxation, counter ion relaxation and Maxwell-Wagner effects). The frequency-dependent HF-EM properties were further modeled based on a novel hydraulic-mechanical-electromagnetic coupling approach developed for soils. The results show the potential of HF-EM techniques for quantitative monitoring of the hydraulic state in underground repositories in clay formations. Key Points Dielectric relaxation behavior of porous mediaRadio and microwave remote sensing techniquesHydraulic-mechanical-electromagnetic couplin

Utilisation des données Google Street View pour cartographier la distribution géographique des espèces. Une étude préliminaire de la processionnaire du pin (Thaumetopoea pityocampa)

Article publié suite à l'événement : MEDINSECT 3 ; Hammamet-Tunis (Tunisie) - (2012-05-08 - 2012-05-11).Mapping species distribution is an important and useful task to monitor invasive species spread or native species expansion under climate change. Unfortunately it requires a lot of occurrence data that are not easily available from literature and that are very time-consuming to collect in the field. For that reason, we designed a survey with the aim to explore to which extent large-scale databases such as Google Street View could be used to derive valid occurrence data. We worked with an insect species, the Pine Processionary Moth (PPM) Thaumetopoea pityocampa because the larvae of that moth build silk nests on its host tree that are easily visible. The presence of the species at one location can therefore be inferred from visual records derived from the panoramic views available from Google Street View. We designed a standardized procedure allowing the evaluation of the presence of the PPM on a large sampling grid (covering 46 848 km2) located in France. Field sampling has been conducted in parallel, which allowed a straightforward comparison between field and Google-derived datasets. Data derived from Google Street View were highly similar to field data as we found an accuracy (percentage of field values correctly predicted using Google Street View) of 92.9%. We conclude that Google database might provide useful occurrence data for mapping the distribution of species which presence can be visually evaluated such as the PPM. More data are needed, however, to assess the range of spatial scales at which Google Street View actually provides reliable occurrence data.La cartographie de la distribution géographique des espèces est importante pour suivre l’évolution des aires de distribution d’espèces invasives ou d’espèces natives en expansion géographique. Malheureusement, les données nécessaires sont parfois difficilement accessibles à partir de la littérature et sont coûteuses à collecter sur le terrain. Pour cette raison, nous avons conçu une étude dans le but d'explorer dans quelle mesure il est possible d’utiliser les bases de données telles que Google Street View (GSV) pour obtenir des données d’occurrence valides. Nous avons choisi de travailler avec une espèce d’insecte, la chenille processionnaire du pin (PP) Thaumetopoea pityocampa car les larves de cette espèce se développent dans le feuillage des arbres hôtes et tissent un nid blanc aisément visible. La présence de l'espèce dans un site donné peut donc être facilement renseignée en examinant les vues panoramiques disponibles pour de nombreuses localités dans la base de données de Google Street View. Nous avons conçu une procédure standardisée permettant d'évaluer la présence de la PP à partir des données GSV et nous l’avons mise en oeuvre sur une aire d’étude couvrant 46 848 km2dans la région Centre en France. La distribution de l’espèce a également été décrite à l’aide d’échantillonnages réalisés sur le terrain. Les données issues de l’examen des images Google Street View ont été comparées aux données de terrain et se sont révélées de bons estimateurs de la présence de la processionnaire du pin avec une précision (proportion de valeurs correctement estimées) de 92.9% sur notre zone d’étude pour un maillage de 16 km x 16 km. Ces résultats suggèrent que l’exploitation des bases de données de GSV pourrait permettre de produire des données écologiques intéressantes pour les espèces dont la présence peut être estimée visuellement à partir de photographies. Des études complémentaires sont cependant nécessaires pour mieux cerner la gamme d’échelles spatiales auxquelles GSV fournit des données d’occurrence fiables

Adaptation of neutrophilic Paracoccus denitrificans to denitrification at highly alkaline pH

Bacterial denitrification is widely documented at neutral pH in order to improve the removal of nitrate in wastewater treatment processes. However, certain industrial contexts generate alkaline waste and effluent containing nitrate that must be denitrified. To obtain more information on denitrification at alkaline pH, this study evaluated the possibility of adapting a neutrophilic denitrifying strain, Paracoccus denitrificans, to alkaline pH. Firstly, P. denitrificans’ denitrifying activity was evaluated without acclimation in batch bioreactors at pH 7.0, 8.0, 9.0 and 10.0. Then, two acclimation methods using successive batch bioreactors and a continuous bioreactor allowed P. denitrificans to be gradually exposed to alkaline pH: from 8.5 to 11.2 in 26 and 72 days respectively. Results showed that P. denitrificans could grow and catalyse nitrate reduction (i) at pH 9.0 without acclimation, (ii) at pH 10.5 in successive batch cultures with progressively increasing pH and (iii) at pH 10.8 in continuously fed culture with a hydraulic retention time (HRT) of 8 days. It was shown that denitrification affected the pH despite the presence of carbonate buffering of the P. denitrificans growth medium. With acetate as an electron donor, the pH of a carbonate buffered medium tends towards pH 10 during the process of denitrification

Nitrate and nitrite reduction activity of activated sludge microcosm in a highly alkaline environment with solid cementitious material

Denitrification is a major biological process contributing to nitrate and nitrite reduction. However, this process remains poorly understood at alkaline pH although such conditions can be encountered in natural (e.g. soda lakes) or industrial environments (e.g. geological waste repositories with cementitious materials). To investigate the nitrate reduction (NR) rate for pH > 9.5 in a cementitious environment, several batch reactors were implemented, with cement leachate or with hardened cement paste (HCP). In the experiments carried out with cement leachate, NR dropped from 0.72 mM/h at pH 9.5 to 0.17 mM/h at pH > 11, while the concentration of nitrite increased. The NR was inhibited at pH close to 12, as was the nitrite reduction at pH above 11. In the reactor containing HCP, the NR rate was 0.75 mM/h at pH close to 10. Calcite precipitated on the HCP surface. Epifluorescence microscopy observations coupled with DNA labelling suggested the presence of microorganisms attached to the HCP surface. This was confirmed by biological growth coupled with NR activity after the transfer of the HCP into a new medium, considered to be sterile. The bacterial community analysis showed that the highly selective culture conditions led to the selection of two species: Halomonas sp. and a species known for its versatile metabolism and ability to form biofilms, i.e. Thauera sp