This study aimed to assess the potential impact of long-term chronic exposure (69 years) to naturally-occurring radionuclides (RNs) and heavy metals on microbial communities in sediment from a stream flowing through a watershed impacted by an ancient mining site (Rophin, France). Four sediment samples were collected along a radioactivity gradient (for 238U368 to 1710 Bq.Kg-1) characterized for the presence of the bioavailable fractions of radionuclides (226Ra, 210Po), and trace metal elements (Th, U, As, Pb, Cu, Zn, Fe). Results revealed that the available fraction of contaminants was significant although it varied considerably from one element to another (0 % for As and Th, 5-59 % for U). Nonetheless, microbial communities appeared significantly affected by such chronic exposure to (radio)toxicities. Several microbial functions carried by bacteria and related with carbon and nitrogen cycling have been impaired. The high values of fungal diversity and richness observed with increasing downstream contamination (H' = 4.4 and Chao1 = 863) suggest that the community had likely shifted toward a more adapted/tolerant one as evidenced, for example, by the presence of the species Thelephora sp. and Tomentella sp. The bacterial composition was also affected by the contaminants with enrichment in Myxococcales, Acidovorax or Nostocales at the most contaminated points. Changes in microbial composition and functional structure were directly related to radionuclide and heavy metal contaminations, but also to organic matter which also significantly affected, directly or indirectly, bacterial and fungal compositions. Although it was not possible to distinguish the specific effects of RNs from heavy metals on microbial communities, it is essential to continue studies considering the available fraction of elements, which is the only one able to interact with microorganisms.

Civilian and military nuclear programs of several nations over more than 70+ years have led to significant quantities of heterogenous solid, organic, and aqueous radioactive wastes bearing actinides, fission products, and activation products. While many physicochemical treatments have been developed to remediate, decontaminate and reduce waste volumes, they can involve high costs (energy input, expensive sorbants, ion exchange resins, chemical reducing/precipitation agents) or can lead to further secondary waste forms. Microorganisms can directly influence radionuclide solubility, via sorption, accumulation, precipitation, redox, and volatilization pathways, thus offering a more sustainable approach to remediation or effluent treatments. Much work to date has focused on fundamentals or laboratory-scale remediation trials, but there is a paucity of information toward field-scale bioremediation and, to a lesser extent, toward biological liquid effluent treatments. From the few biostimulation studies that have been conducted at legacy weapon production/test sites and uranium mining and milling sites, some marked success via bioreduction and biomineralisation has been observed. However, rebounding of radionuclide mobility from (a) biotic scale-up factors are often encountered. Radionuclide, heavy metal, co-contaminant, and/or matrix effects provide more challenging conditions than traditional industrial wastewater systems, thus innovative solutions via indirect interactions with stable element biogeochemical cycles, natural or engineered cultures or communities of metal and irradiation tolerant strains and reactor design inspirations from existing metal wastewater technologies, are required. This review encompasses the current state of the art in radionuclide biogeochemistry fundamentals and bioremediation and establishes links toward transitioning these concepts toward future radioactive effluent treatments.

Some natural environments on Earth are characterised by high levels of radiation, including naturally radioelement enriched mineral springs in the French Massif Central. Therefore, naturally radioactive mineral springs are interesting ecosystems for understanding how bacterial populations in these springs have adapted to high levels of natural and chronic radioactivity over the very long term. The aim of this study was to analyse the bacterial communities of sediments from five naturally radioactive mineral springs in the French Massif Central, sampled in autumn 2019 and spring 2020, and to observe whether radionuclides, compared to other physicochemical parameters, are drivers of the bacterial community structuring in these extreme environments. Physicochemical measurements showed that two springs, Dourioux and Montagne had high radioelement concentrations/activities (uranium, thorium and radon). Analysis of the structure of the bacterial communities, by next generation sequencing based on 16S rRNA gene sequencing, showed that the presence of radionuclides in Dourioux and Montagne, did not lead to a reduction in bacterial diversity and richness compared to the other springs. However, Dourioux and Montagne were characterised by specific bacterial populations, whose presence correlates with the radioelement concentrations/activities measured in these springs. This suggests that radioelements could partly explain the structuring of bacterial communities in these springs. In addition, several of these operational taxonomic units (OTUs) specific to Dourioux and Montagne, mainly affiliated to Proteobacteria, Firmicutes, Acidobacteria, Actinobacteria, and Bacteroidetes, could be involved in the biogeochemistry of radionuclides through different mechanisms (biosorption, biomineralisation, bioaccumulation, and bioreduction), which would allow the development of other bacterial species sensitive to these metals/radioelements. In particular, the co-occurrence of sulphate and/or iron-reducing bacteria, capable of bioreducing uranium, with fermentative bacteria, releasing sources of organic carbons, reflects associations of bacteria with complementary functions that allow them to grow in this peculiar environment and maintain a high diversity in these extreme environments. This study has provided a better understanding of the structuring of bacterial communities exposed to ionising radiation for thousands of years in naturally radioactive environments.

The TIRAMISU collaboration gathers expertise from biologists, physicists, radiochemists and geologists within the Zone-Atelier Territoires Uranifères (ZATU) in France to analyze the radiation exposure of microorganisms living in naturally radioactive mineral springs. These springs are small waterbodies that are extremely stable over geological time scales and display different physicochemical and radiological parameters compared to their surroundings. Water and sediment samples collected in 27 mineral springs of the volcanic Auvergne region (Massif Central, France) have been studied for their microbial biodiversity and their radionuclide content. Among the microorganisms present, microalgae (diatoms), widely used as environmental indicators of water quality, have shown to display an exceptional abundance of teratogenic forms in the most radioactive springs studied (radon activity up to 3700 Bq/L). The current work presents a first assessment of the dose received by the diatoms inhabiting these ecosystems. According to ERICA tool, microorganisms living in most of the sampled mineral springs were exposed to dose rates above 10 μGy/h due to the large concentration of radium in the sediments (up to 50 Bq/g). Radiological analyses of water and sediments were used as inputs to Monte Carlo simulations at micro-(GATE) and nano- (Geant4-DNA) scale in order to assess the direct and indirect damages on the diatom DNA.

Although many studies have explored the responses of living organisms or ecosystems exposed to radioelements from human activities, very few studies have focused on the characterization of microbial biodiversity and the adaptations developed in environments characterized by natural, chronic and old radioactivity. Radioactive mineral springs are insulated ecosystems that have barely evolved in the last centuries. They therefore offer exceptional windows on the history of life on earth. Within the framework of the TIRAMISU project (biodiversiTy In RAdioactive MIneral Springs of the aUvergne region), biologists, ecologists, geologists, physicists, radiochemists and sociologists explore if and how natural ionizing radiation can be an “abiotic driver” impacting the diversity and structuring of microbial communities in food webs exposed to this radiation for long periods.

In 2008, as part of a feasibility study for radioactive waste disposal in deep geological formations, the French National Radioactive Waste Management Agency (ANDRA) drilled several boreholes in the transposition zone in order to define the potential variations in the properties of the Callovo–Oxfordian claystone formation. This consisted of a rare opportunity to investigate the deep continental biosphere that is still poorly known. Four rock cores, from 1709, 1804, 1865, and 1935 m below land surface, were collected from Lower and Middle Triassic formations in the Paris Basin (France) to investigate their microbial and geochemical composition. Rock leachates showed high salinities ranging from 100 to 365 g·L−1 NaCl, current temperatures averaging 65 °C, no detectable organic matter, and very fine porosity. Microbial composition was studied using a dual cultural and molecular approach. While the broad-spectrum cultural media that was used to activate microbial communities was unsuccessful, the genetic investigation of the dominant 16S rRNA gene sequences revealed eight bacterial genera considered as truly indigenous to the Triassic cores. Retrieved taxa were affiliated to aerobic and facultative anaerobic taxon, mostly unknown to grow in very saline media, except for one taxon related to Halomonas. They included Firmicutes and α-, β-, and γ-Proteobacteria members that are known from many subsurface environments and deep terrestrial and marine ecosystems. As suggested by geochemical analyses of rocks and rock leachates, part of the indigenous bacterial community may originate from a cold paleo-recharge of the Trias aquifer with water originating from ice melting. Thus, retrieved DNA would be fossil DNA. As previously put forward to explain the lack of evidence of microbial life in deep sandstone, another hypothesis is a possible paleo-sterilisation that is based on the poly-extremophilic character of the confined Triassic sandstones, which present high salinity and temperature.

After the Chernobyl nuclear power plant accident in 1986, contaminated soils, vegetation from the Red Forest and other radioactive debris were buried within trenches. In this area, trench T22 has long been a pilot site for the study of radionuclide migration in soil. Here, we used 454 pyrosequencing of 16S rRNA genes to obtain a comprehensive view of the bacterial and archaeal diversity in soils collected inside and in the vicinity of the trench T22 and to investigate the impact of radioactive waste disposal on prokaryotic communities. A remarkably high abundance of Chloroflexi and AD3 was detected in all soil samples from this area. Our statistical analysis revealed profound changes in community composition at the phylum and OTUs levels and higher diversity in the trench soils as compared to the outside. Our results demonstrate that the total absorbed dose rate by cell and, to a lesser extent the organic matter content of the trench, are the principal variables influencing prokaryotic assemblages. We identified specific phylotypes affiliated to the phyla Crenarchaeota, Acidobacteria, AD3, Chloroflexi, Proteobacteria, Verrucomicrobia and WPS-2, which were unique for the trench soils.

Although uranium (U) is naturally found in the environment, soil remediation programs will become increasingly important in light of certain human activities. This work aimed to identify U(VI) detoxification mechanisms employed by a bacteria strain isolated from a Chernobyl soil sample, and to distinguish its active from passive mechanisms of interaction. The ability of the Microbacterium sp. A9 strain to remove U(VI) from aqueous solutions at 4 °C and 25 °C was evaluated, as well as its survival capacity upon U(VI) exposure. The subcellular localisation of U was determined by TEM/EDX microscopy, while functional groups involved in the interaction with U were further evaluated by FTIR; finally, the speciation of U was analysed by TRLFS. We have revealed, for the first time, an active mechanism promoting metal efflux from the cells, during the early steps following U(VI) exposure at 25 °C. The Microbacterium sp. A9 strain also stores U intracellularly, as needle-like structures that have been identified as an autunite group mineral. Taken together, our results demonstrate that this strain exhibits a high U(VI) tolerance based on multiple detoxification mechanisms. These findings support the potential role of the genus Microbacterium in the remediation of aqueous environments contaminated with U(VI) under aerobic conditions.

The diversity of bacterial communities exposed to radioactive contamination in Chernobyl soils was examined by a combination of molecular and culture-based approaches. A set of six radioactive soil samples, exhibiting high levels of Cs-137 contamination, were collected from the T22 trench. Three samples were also collected in nearby soils with low contamination. Complex bacterial community structures were observed in both highly and weakly contaminated samples, using a molecular approach targeting the 16S rRNA gene. However, the presence of specific populations within samples from highly contaminated soils could not be revealed by statistical analysis of the DGGE profiles. More than 200 culturable isolates, representative of dominant morphotypes, were grouped into 83 Operational Taxonomic Units (OTUs) and affiliated to Firmicutes, Actinobacteria, Alpha-, Beta-, Gamma-Proteobacteria and Bactero detes. No specific pattern linked to contamination was observed for these culturable bacteria. The results show that both highly and weakly contaminated soils host a wide diversity of bacteria, suggesting that long term exposure to radionuclides does not lead to the extinction of bacterial diversity. (C) 2011 Elsevier Ltd. All rights reserved.

Steroid hormones are essential signalling molecules in prostate cancer (PC). However, many studies focusing on liquid biomarkers fail to take the hormonal status of these patients into account. Steroid measurements are sensitive to bias caused by matrix effects, thus assessing potential matrix effects is an important step in combining circulating tumour DNA (ctDNA) analysis with hormone status.We investigated the accuracy of multi-steroid hormone profiling in mechanically-separated plasma (MSP) samples and in plasma from CellSave Preservative (CS) tubes, that are typically used to obtain ctDNA, compared to measurements in serum. We performed multiplex steroid profiling by liquid chromatography–tandem mass spectrometry (LC–MS/MS) in samples obtained from ten healthy controls and ten castration-resistant prostate cancer (CRPC) patients.Steroid measurements were comparable between MSP and serum. A small but consistent decrease of 8–21% compared to serum was observed when using CS plasma, which was considered to be within the acceptable margin. The minimal residual testosterone levels of CRPC patients could be sensitively quantified in both MSP and CS samples.We validated the use of MSP and CS samples for multi-steroid profiling by LC–MS/MS. The optimised use of these samples in clinical trials will allow us to gain further insight into the steroid metabolism in PC patients.

The purpose of the Porewater Chemistry (PC) experiment at the Mont Terri (MT) Underground Rock Laboratory (URL) was to measure geochemical parameters, such as pH, Eh and pCO(2), in the porewater of the Opalinus Clay formation. Although the PC experiment was designed and implemented carefully from a geochemical perspective, conditions were not sterile and some microbial and nutrient contamination likely occurred. Microbial activity in the added synthetic porewater in the borehole was apparent shortly after initiation of the experiment and affected the geochemical parameters observed in the porewater. This paper summarizes the results from microbial analyses of post-termination PC water and overcore clay samples, conducted to attempt to elucidate the role of microbial activity in the evolution of the geochemical conditions in the PC experiment. Microbial analyses of the PC borehole water, and of clay overcore samples from around the borehole, were carried out at three laboratories and included both molecular biology and culturing methods.Results indicated the presence of heterotrophic aerobic and anaerobic organisms that resulted likely from the initial, non-sterile conditions, sustained by suspected contamination with organic matter (glycerol, acetone). The results also indicated the presence of NO(3)-reducers, Fe-reducers, SO(4)-reducers and methanogens (i.e.. Bacteria as well as Archaea), suggesting a reducing environment with Fe(III)- and SO(4) reduction, and methanogenesis occurring in the PC water and adjacent clay. A black precipitate containing pyrite (identified by XRD and SEM) and a strong H(2)S smell in the porewater confirmed the occurrence of SO(4) reduction. Microorganisms identified in the porewater included Pseudomonas stutzeri, Bacillus licheniformis, Desulfosporosinus spp. and Hyphomonas spp. Species identified in enrichment cultures from the overcore samples included Pseudomonas stutzeri, three species of Trichococcus spp., Caldanaerocella colombiensis, Geosporobacter subterrenus and Desulfosporosinus locus. Overall the results indicated a thriving microbial community in the PC water and adjacent clay in contrast to "undisturbed" Opalinus Clay for which limited evidence for a small viable microbial community has been given in a previous study. Crown Copyright (C) 2011 Published by Elsevier Ltd. All rights reserved.