Deep Geological Environment Research Articles

Microfluidics for studying the deep underground biosphere: from applications to fundamentals.

In this review, selected examples are presented to demonstrate how microfluidic approaches can be utilized for investigating microbial life from deep geological environments, both from practical and fundamental perspectives. Beginning with the definition of the deep underground biosphere and the conventional experimental techniques employed for these studies, the use of microfluidic systems for accessing critical parameters of deep life in geological environments at the microscale is subsequently addressed (high pressure, high temperature, low volume). Microfluidics can simulate a range of environmental conditions on a chip, enabling rapid and comprehensive studies of microbial behavior and interactions in subsurface ecosystems, such as simulations of porous systems, interactions among microbes/microbes/minerals, and gradient cultivation. Transparent microreactors allow real-time, noninvasive analysis of microbial activities (microscopy, Raman spectroscopy, FTIR microspectroscopy, etc.), providing detailed insights into biogeochemical processes and facilitating pore-scale analysis. Finally, the current challenges and opportunities to expand the use of microfluidic methodologies for studying and monitoring the deep biosphere in real time under deep underground conditions are discussed.

Research progress on microbial adsorption of radioactive nuclides in deep geological environments.

Due to the development and utilization of nuclear energy, the safe disposal of nuclear waste needs to be urgently addressed. In recent years, the utilization of microorganisms' adsorption capacity to dispose of radioactive waste has received increasing attention. When compared with conventional disposal methods, microbial adsorption exhibits the characteristics of high efficiency, low cost, and no secondary pollution. In the long term, microbial biomass shows significant promise as specific chemical-binding agents. Optimization of biosorption conditions, identification of rare earth element binding sites, and studies on the sorption capacities of immobilized cells provide compelling reasons to consider biosorption for industrial applications in heavy metal removal from solutions. However, the interaction mechanism between microorganisms and radioactive nuclides is very complex. This mini-review briefly provides an overview of the preparation methods, factors affecting the adsorption capacity, and the mechanisms of microbial adsorbents.

Effect of Iron Chloride (II) on Bentonites under Hydrothermal Gradients: A Comparative Study between Sodium Bentonite and Calcium Bentonite

This study investigates the performance of two bentonite materials, specifically MX-80 (Na-bentonite) and FEBEX (Ca-Mg-Na-bentonite), employed as engineered barriers in deep geological disposal facilities for the isolation of high-level radioactive waste, contained in metallic canisters. Experiments conducted at the laboratory scale focused on the interaction of these bentonites with FeCl2 powder, used as a soluble iron source, to observe enhanced alteration of the bentonite. The experiments were carried out under a hydrothermal gradient. A dominant Na-Cl-SO4 saline solution was put in contact with the compacted bentonites from the top, while a constant temperature of 100 °C was maintained at the bottom using a heater in contact with the layer of FeCl2. The experimental cells were examined after six months of interaction. Various changes in the physical and chemical properties of the bentonites were observed. An increase in the water content, a reduction in the specific surface area and cation exchange capacity, changes in the distribution of aqueous species, and the formation of secondary minerals were observed. Reaction products formed at the bentonite interface with FeCl2, primarily comprising akaganeite, goethite, and hematite. The smectites showed evident structural modifications, with an enrichment in iron content, and a shift in the exchangeable ion distribution in the case of MX-80 bentonite. This work provides valuable insights into the complex interactions between bentonite barriers and materials that dissolve iron, serving as proxies for deep geological disposal environments and indicating the potential long-term behavior, taking into account higher concentrations of dissolved iron than those expected in a real repository.

A comparative analysis of the performance of backfill heat exchangers in deep mine geological environments

With the increase of mineral mining depth, the technologies of constructing heat exchanger in stopes to exploit geothermal resources from deep mines have received great attention. The heat extraction capacities of heat exchangers are closely related to the geological environments of deep mines, especially backfill mines. Therefore, a three-dimensional unsteady heat transfer and seepage coupling model of backfill heat exchangers (BFHEs) is created and verified in this work using COMSOL simulation software. Then, the influences of geological environments, involving groundwater seepage, initial temperature and thermal conductivity of surrounding rock etc., on the ability of five BFHEs with the equal buried tube length to extract heat in different configurations are investigated by this model. The findings indicate that groundwater flow can significantly improve the performance of BFHEs. Double-layer tube BFHEs have the best overall performance in the studied seepage velocity range. Only when there is no or little seepage is the performance of single-layer tube BFHEs equivalent to that of double-layer tube BFHEs. The heat extraction capacity of BFHEs is greatly improved under seepage conditions with the increase in the initial temperature of surrounding rock, although the impact is low in the absence of groundwater seepage. The benefit of backfill body's thermal conductivity on BFHEs' ability to transport heat is significantly greater than that of surrounding rock, but the effect becomes smaller as the value increases. Further increasing the thermal conductivity of the backfill body will not considerably increase the heat extraction capability of BFHEs after it reaches 2.5 (W·m−1·K−1). Therefore, if the cost is too high, continued enhancement is not recommended. The findings offer theoretical recommendations for optimizing BFHEs’ setup to better adapt to deep mine geological environments and increase geothermal extraction capability.

Effect of hydrogen gas and leaching solution on the fast release of fission products from two PWR fuels

To study the dissolution of UOX spent nuclear fuel in a deep geological environment and the fast release of a selection of relevant radionuclides for long-term safety of this high level waste, leaching experiments were performed with spent nuclear fuel samples originating from the pressurized water reactors (PWRs) Tihange 1 and Gösgen with a similar burnup (50 – 55 MWd.kgHM−1) but different irradiation histories. Six experiments were conducted to investigate the effect of two critical parameters: (1) the highly alkaline environment caused by the presence of cementitious materials in the “Supercontainer design”, which is currently the reference design for the long-term management of the high-level nuclear waste forms in Belgium, and (2) the reducing conditions imposed by the presence of hydrogen from the corrosion of iron-based materials present in the engineered barriers. The experiments were performed using autoclaves under pressure from 1 to 40 bar with a pure Ar atmosphere or a mixture of H2/Ar. Divided into two consecutive phases, the total experimental duration was about 1400 days. The Phase I provided mainly information about the fast release of the fission products while the perspective of the Phase II was to study the long-term evolution of the spent fuel matrix. During the leaching experiment, concentrations of a selection of radionuclides (238U, 129I, 137Cs, 90Sr and 99Tc) were monitored in solution and the amounts of Kr and Xe were measured in the gas phase. Based on results of the experiments conducted for up to 40 months (i.e. during Phase I of the experimental program), we observe that there is a continuous release of 137Cs, 90Sr and of the fission gases and a clear impact of the irradiation history on the release of certain fission products.

The corrosion behavior of nonferrous metals in deep geological repository environments

AbstractThe advanced technologies of modern civilization produce radioactive wastes that require careful disposal if they cannot be recycled. These materials can originate from a variety of activities, such as scientific research, medicine, or nuclear power generation and, as such, can result in numerous waste forms. In this paper, the corrosion behavior of several less‐common metals is studied, specifically: aluminum, copper, lead, magnesium, zinc, and zirconium, all under simulated cementitious environments. The data reported rely on the production of hydrogen as a corrosion end‐product to calculate the uniform corrosion rate as a function of time. At 50°C, in either young cement water (pH 13.5) or saturated portlandite (pH 12.5) and after approximately 2 years of exposure, magnesium was found to corrode at ∼10 µm/year; aluminum at 1 µm/year (portlandite only); zinc at ∼100 nm/year; lead at <1 nm/year and both copper and zirconium at less than 0.1 nm/year.

Assessing the tunnel stability in brittle rocks based on strain bursting assessment

Strain bursting is a phenomenon associated with deep underground excavations and tunnelling in brittle hard rocks that result in a sudden and violent failure, posing high risks and damage. This work aims to assess the strain bursting and the energy response characteristics and develop a guideline that can be utilized in the preliminary investigation stage. The obtained results are intended to enable predicting different brittle behaviours in deep geological environments. Elastic stress analysis using the DISL approach based on boundary element method is established (EX3-software) for the assessment of two tunnel geometries (circular, horseshoe) in granitic rock mass. Two different stress states are modelled to simulate the potential stress regime. The analysis’ results demonstrate that different geometries affect the energy storage capacity and result in different behaviour regarding the bursting potential of the rock mass.

Genesis of Significance of Carbonated Thermal Water Springs in Xining Basin, China

There are 30 carbonate hot springs in Yaoshuitan geothermal field, Xining Basin, China, with a temperature of 18~41.5 °C; and there are 10 carbonate hot springs in Qijiachuan geothermal field, with a temperature of 10~19.5 °C. Both geothermal fields are carbonate hot springs containing large amounts of CO2 gas. In order to reveal the origin of the carbonated hot springs in Yaoshuitan and Qijiachuan of Xining Basin, this paper offers a comprehensive study of the regional deep geology, tectonic setting, total analysis of carbonated hot springs, δ2H, δ18O, δ13C isotopes, main gas composition, and geochemical characteristics of travertine dating, travertine δ13C, and rare earth elements. The geological process of carbonated hot spring formation and the evolution of H+ content from deep to shallow is revealed, and the genetic mechanism of the carbonated hot spring in Xining Basin is systematically summarized. The results show that: (1) The characteristics of δ2H and δ18O isotopes indicate that the recharge source of carbonated thermal water springs in Xining Basin is mainly atmospheric precipitation. The age of carbonated thermal water springs at 14C is more than 20 ka, indicating that some of them may come from deep fluid (gas) sources. The R/Ra in carbonated thermal water springs is mostly less than 1, indicating that the helium in geothermal water is mainly crustal source helium, and there is no deep mantle source material. (2) The Piper three-plot indicates that the direction of groundwater evolution from the recharge area at the edge of Xining Basin to Yaoshuitan and Qijiachuan carbonated thermal water spring area near the edge of the basin is opposite to the normal path of groundwater evolution in the basin, which is due to the large amount of CO2 gas mixed in the deep fault along the northern margin of Laji Mountain. The ratio of (Ca2+ + Mg2+) and (HCO3− + SO42−) in the Potan and Qijiachuan carbonated thermal water springs is close to 1, and the ratio of (Na+ + K+)/HCO3− is less than 1. It indicates that the chemical composition of the Yaoshuitan carbonated thermal water spring and the Qijiachuan carbonated thermal water spring in Xining Basin is dominated by the dissolution of calcite, dolomite, and gypsum in deep carbonate reservoirs, supplemented by the dissolution of silicate minerals. The relationship between the volume fraction of CO2 and the δ13C value of carbon isotope of CO2 indicates that the source of CO2 is inorganic, which is mainly formed by metamorphism and decomposition of deep carbonate and marble. The δEu < 1 and δCe > 1 of the rare earth elements in the calcium center of the carbonated thermal water springs indicate that the groundwater supplying the travertine material has been in the acidic environment receiving CO2 from the deep crust for a long time. (3) A series of tectonic activities, such as late collision and post-collision between the Indian and Eurasian plates, has led to the uplift, asthenosphere upwelling, and thermal invasion of the northern Tibetan Plateau and other deep dynamic processes. The deep faults in the northern margin of the Laji Mountain and other deep faults with obvious neotectonic activity have provided channels for the up-invasion of deep thermal materials, and local geothermal anomalies were formed near the deep faults. The hidden carbonate rocks and silicate rocks with large thickness undergo thermal metamorphism under high temperature and high pressure in the deep geothermal anomaly area and form a large amount of CO2, which is dissolved in water and enhances the acidity of water. At the same time, the dissolution reaction of acidic water to carbonate rocks consumes H+, which keeps the carbonated thermal water spring weakly acidic. (4) The composition of travertine in carbonated thermal water springs is dominated by calcite, indicating that travertine may be formed in a deep geological environment with a temperature of 150~200 °C, indicating that there are abnormal heat sources in shallow carbonate strata with a burial depth of 3000~4000 m. The abnormal heat source may be caused by the deep fault in the northern margin of Laji Mountain, as well as other deep and large faults channeled in the deep crust and mantle heat source, indicating that the deep fault in the northern margin of Laji Mountain has an obvious heat-controlling effect, and there is a good prospect of geothermal resources exploration near the fault.

Multi-level voxel representations for digital twin models of tunnel geological environment

• An efficient linear segmentation-based multi-level voxel representation method is proposed, aiming to represent the geological environment of the long and deep tunnel. Based on the results of the comparative experiment, the spatial efficiency was augmented by 30% approximately by using this technique. • VDB data structure is used to meet the dynamic update requirement of the digital twin applications. It has the consistent time complexity of random access and uses a narrow-band level set to reduce space complexity. • Multi-source data is integrated to represent the multi-feature geological environment. Four typical data with different modalities (parametric BIM model, B-rep solid model, triangle mesh model, 3D regular grid model) are successfully integrated into the digital twin model. Intelligent tunnel engineering requires accurate and comprehensive digital twin models to represent complex geological environments. The digital twin model of tunnel geological environment has multi-level and diversified in-depth applications such as problem diagnosis, risk assessment, trend prediction, emergency response, etc. Since tunnel construction is a long-term continuous dynamic spatial–temporal progress, the tunnel face constantly advances during excavations. Thus, the geological conditions of the construction surface and surrounding areas are continuously revealed and updated, which causes the digital twin model to have a very high update frequency of the geological information in tunnel engineering. The existing methods fail to represent and update the geological environment comprehensively and in real-time, especially for those long deep tunnels with the complicated topology of geological structures and the heterogeneous distribution of internal properties. Due to the digital twin application having a high-level requirement of completeness, accuracy, and timeliness, 3D modeling of tunnel geological environments has remained challenging within digital twin application contexts. This study aims to deliver a solution that efficiently represents and updates the geological structures and internal non-uniform properties of tunnel engineering. Since dense voxels can represent inner intricate heterogeneous property information, sparse voxels can efficiently represent complex structural information. Meanwhile, voxels using Volumetric Dynamic B + trees (VDB) data structure are easy to integrate and update. However, according to the characteristics of long strip distribution of tunnel geological environments, directly using the VDB will still cause an efficiency problem. Therefore, this study proposes an efficient linear segmentation-based multi-level voxel representation method for the prolonged deep tunnel geological environment using the VDB data structure to support dynamic updates. The typical Mountain Tunnel of China is employed for experimental analysis to validate the proposed method, and four representative data with different modalities are integrated into the digital twin model. The results demonstrate that spatial efficiency augments 28.49% after segmentation, and data access with O(1) time complexity supports efficiently dynamic updates.

Evolution Characteristics of Overburden Instability and Failure under Deep Complex Mining Conditions

Aiming at the problems of intensified deformation and serious stress damage of stope overburden during mining in deep typical geological environment, taking the deep mining face of a mine in the northwest as the research background, the characteristics and influencing factors of overburden collapse in deep geological environment are analyzed, a comprehensive evaluation system affecting overburden instability and deformation is established, and the temporal and spatial evolution law of deep stope roof failure is obtained by using the “three-zone” distribution theory of overburden, analytic hierarchy process, and numerical simulation method. The results show that high ground stress has the most significant impact on the geological environment of deep stope, followed by the properties of surrounding rock, hydrogeological conditions, and unloading disturbance. The damage degree of the plastic zone at the top of the overburden is greater than that at the bottom. With the increase of mining time, the damage range of the roof and floor gradually increases. In the process of coal seam mining, the increasing range and sinking trend of roof stress are larger near the working face, and the farther away from the working face, the increasing range decreases continuously. The research conclusion provides an important reference for coal seam safety mining and surrounding rock control in complex environments such as deep mining, high stress, and strong disturbance.

Biogeochemical Cycling of 99Tc in Alkaline Sediments.

99Tc will be present in significant quantities in radioactive wastes including intermediate-level waste (ILW). The internationally favored concept for disposing of higher activity radioactive wastes including ILW is via deep geological disposal in an underground engineered facility located ∼200-1000 m deep. Typically, in the deep geological disposal environment, the subsurface will be saturated, cement will be used extensively as an engineering material, and iron will be ubiquitous. This means that understanding Tc biogeochemistry in high pH, cementitious environments is important to underpin safety case development. Here, alkaline sediment microcosms (pH 10) were incubated under anoxic conditions under "no added Fe(III)" and "with added Fe(III)" conditions (added as ferrihydrite) at three Tc concentrations (10-11, 10-6, and 10-4 mol L-1). In the 10-6 mol L-1 Tc experiments with no added Fe(III), ∼35% Tc(VII) removal occurred during bioreduction. Solvent extraction of the residual solution phase indicated that ∼75% of Tc was present as Tc(IV), potentially as colloids. In both biologically active and sterile control experiments with added Fe(III), Fe(II) formed during bioreduction and >90% Tc was removed from the solution, most likely due to abiotic reduction mediated by Fe(II). X-ray absorption spectroscopy (XAS) showed that in bioreduced sediments, Tc was present as hydrous TcO2-like phases, with some evidence for an Fe association. When reduced sediments with added Fe(III) were air oxidized, there was a significant loss of Fe(II) over 1 month (∼50%), yet this was coupled to only modest Tc remobilization (∼25%). Here, XAS analysis suggested that with air oxidation, partial incorporation of Tc(IV) into newly forming Fe oxyhydr(oxide) minerals may be occurring. These data suggest that in Fe-rich, alkaline environments, biologically mediated processes may limit Tc mobility.

Research Progress on Hydrogen Absorption and Embrittlement of Titanium and Its Alloy for High-level Nuclear Waste Container in Deep Geological Disposal Environment

Research Progress on Hydrogen Absorption and Embrittlement of Titanium and Its Alloy for High-level Nuclear Waste Container in Deep Geological Disposal Environment

Analyses on Thermal Stability and Structural Integrity of the Improved Disposal Systems for Spent Nuclear Fuels in Korea

With respect to spent nuclear fuels, disposal containers and bentonite buffer blocks in deep geological disposal systems are the primary engineered barrier elements that are required to isolate radioactive toxicity for a long period of time and delay the leakage of radio nuclides such that they do not affect human and natural environments. Therefore, the thermal stability of the bentonite buffer and structural integrity of the disposal container are essential factors for maintaining the safety of a deep geological disposal system. The most important requirement in the design of such a system involves ensuring that the temperature of the buffer does not exceed 100℃ because of the decay heat emitted from high-level wastes loaded in the disposal container. In addition, the disposal containers should maintain structural integrity under loads, such as hydraulic pressure, at an underground depth of 500 m and swelling pressure of the bentonite buffer. In this study, we analyzed the thermal stability and structural integrity in a deep geological disposal environment of the improved deep geological disposal systems for domestic light-water and heavy-water reactor types of spent nuclear fuels, which were considered to be subject to direct disposal. The results of the thermal stability and structural integrity assessments indicated that the improved disposal systems for each type of spent nuclear fuel satisfied the temperature limit requirement (< 100℃) of the disposal system, and the disposal containers were observed to maintain their integrity with a safety ratio of 2.0 or higher in the environment of deep disposal.

Uncovering the electrochemical interface of low-index copper surfaces in deep groundwater environments

Using a combination of a sophisticated modeling protocol and well-established electrochemical techniques, we unravel the chemical composition of the low-index surfaces of copper in groundwater environments at different ion concentrations, pHs, and redox potentials. By carefully linking density functional theory (DFT) and cyclic voltammetry (CV), we are able to extract fundamental information on interfaces of broad significance. Herein, we focus on the case of groundwater found in deep geological environments of importance to the planned constructions of disposal repositories for spent nuclear fuel around the world. Within the error margins of DFT, we can assign adsorption structures and compositions to the current peaks of the CVs. It is found that among the groundwater ions of main interest (i.e. sulfide, bisulfide, sulfate, chloride and bicarbonate), sulfides (HS−, S2−) bind strongest to the surface, and are likely to dominate at the interfaces under the deep geological conditions relevant for repositories of spent nuclear fuel.

Sorption behavior of selenide on montmorillonite

Batch sorption experiments were performed to investigate the sorption mechanism of Se on montmorillonite under reducing conditions in deep geological environments. Based on Eh–pH diagrams and ultraviolet–visible spectra, Se was dissolved as selenide (Se(–II)) anions under the experimental conditions. The distribution coefficients (Kd; m3 kg−1) of Se(–II) indicated ionic strength independence and slight pH dependence. The Kd values of Se(–II) were higher than those of Se(IV), which also exists as an anionic species. X-ray absorption near edge spectroscopy showed that the oxidation state of Se-sorbed on montmorillonite was zero even though selenide remained in the solution. These results suggest that Se(–II) was oxidized and precipitated on the montmorillonite surface. Therefore, it is implied that a redox reaction on the montmorillonite surface contributed to high Kd values for Se(–II).

Effects of deep geological environments for nuclear waste disposal on the hydrogen entry into titanium

Abstract The surface of nuclear waste container will be completely wetted by the groundwater infiltrating through the bentonite or concrete buffer after the closure of its deep geological disposal. Because of its excellent corrosion resistance, titanium is one of the candidates for the container materials for high-level nuclear waste (HLNW) disposal. However, once oxygen depleted in the large time scale deep geological disposal, titanium container will suffer from hydrogen embrittlement (HE). Therefore, in order to predict the potential danger of the HLNW container caused by HE, the hydrogen entry into titanium was studied at different temperatures in simulated deep geological environment of Beishan, which is the preselected HLNW disposal area in China. Devanathan-Stachurski cell methods were firstly implemented in measuring apparent hydrogen diffusivity in titanium with hydrides being formed during hydrogen charging at low temperature ranges (298–373 K). Given the characteristic of the Devanathan-Stachurski cell method's capable of getting apparent diffusivity with combined influencing factors easily, the technique provides a simple way to measure the parameter that is necessary for the estimation of hydrogen penetration depth, facilitating the design of HLNW containers.

Modelling of radiolytic production of HNO3 relevant to corrosion of a used fuel container in deep geologic repository environments

ABSTRACTCopper-coated steel containers are part of the engineered barrier system to permanently store Canadian nuclear fuel waste in a deep geological repository. This work models the dose rates (DRs) at the container surfaces as a function of fuel age. It also utilises a humid-air radiolysis model to study the effects of DR and humidity on radiolytic oxidant production for conditions where unexpected early water intrusion reaches clay seal materials. Radiolysis of humid air produces HNO3. The HNO3 production rate in a condensed water droplet formed on a container surface was conservatively estimated by assuming that every •OH produced by primary radiolytic processes was immediately converted to HNO3 in the gas phase and that all of the HNO3 was absorbed in the water droplet. Also assuming that all of the nitric acid absorbed in the water droplet is consumed in corroding copper and using a hemispherical water droplet geometry, the corrosion depth of the copper coating induced by humid-air radiolysis is conservatively estimated to be 9.4 μm over the permanent storage time.This paper is part of a supplement on the 6th International Workshop on Long-Term Prediction of Corrosion Damage in Nuclear Waste Systems.

Impact of pore fluid salinity on the mechanical behavior of unsaturated bentonite–sand mixture

High salinity of local groundwater and the thermal gradient present within the deep geological repository environment make it necessary to investigate the effect of the pore fluid salinity on the hydro-mechanical behavior engineered barriers at different degrees of saturation. This study examines the impact of CaCl2 salt solution at two concentrations, i.e., 100 and 250 g/L on the compressibility and shear strength behavior of the bentonite–sand buffer (BSB) within a range of saturations. Both as-compacted and desiccated BSB specimens were studied. Desiccated specimens were prepared in a constant-suction environment using the vapor equilibrium technique, and high-pressure cells were used to carry out the triaxial shear experiments. Experimental results were compared with the available data from the literature for BSB materials prepared with distilled water in order to get a better understanding of the impact of the salinity. The comparison shows that the existence of salt in the specimens has notable effect on the total suction levels, compressibility and shear strength of the material.

Geomicrobiology and Metagenomics of Terrestrial Deep Subsurface Microbiomes.

Fractures in the deep subsurface of Earth's crust are inhabited by diverse microbial communities that participate in biogeochemical cycles of the Earth. Life on Earth, which arose c. 3.5-4.0 billion years ago, reaches down at least 5 km in the crust. Deep mines, caves, and boreholes have provided scientists with opportunities to sample deep subsurface microbiomes and to obtain information on the species diversity and functions. A wide variety of bacteria, archaea, eukaryotes, and viruses are now known to reside in the crust, but their functions are still largely unknown. The crust at different depths has varying geological composition and hosts endemic microbiomes accordingly. The diversity is driven by geological formations and gases evolving from deeper depths. Cooperation among different species is still mostly unexplored, but viruses are known to restrict density of bacterial and archaeal populations. Due to the complex growth requirements of the deep subsurface microbiomes, the new knowledge about their diversity and functions is mostly obtained by molecular methods, eg, meta'omics'. Geomicrobiology is a multidisciplinary research area combining disciplines from geology, mineralogy, geochemistry, and microbiology. Geomicrobiology is concerned with the interaction of microorganisms and geological processes. At the surface of mineralogical or rock surfaces, geomicrobial processes occur mainly under aerobic conditions. In the deep subsurface, however, the environmental conditions are reducing and anaerobic. The present chapter describes the world of microbiomes in deep terrestrial geological environments as well as metagenomic and metatranscriptomic methods suitable for studies of these enigmatic communities.

Measuring microbial metabolism in atypical environments: Bentonite in used nuclear fuel storage

Genomics enjoys overwhelming popularity in the study of microbial ecology. However, extreme or atypical environments often limit the use of such well-established tools and consequently demand a novel approach. The bentonite clay matrix proposed for use in Deep Geological Repositories for the long-term storage of used nuclear fuel is one such challenging microbial habitat. Simple, accessible tools were developed for the study of microbial ecology and metabolic processes that occur within this habitat, since the understanding of the microbiota–niche interaction is fundamental to describing microbial impacts on engineered systems such as compacted bentonite barriers. Even when genomic tools are useful for the study of community composition, techniques to describe such microbial impacts and niche interactions should complement these. Tools optimised for assessing localised microbial activity within bentonite included: (a) the qualitative use of the resazurin–resorufin indicator system for redox localisation, (b) the use of a CaCl2 buffer for the localisation of pH, and (c) fluorometry for the localisation of precipitated sulphide. The use of the Carbon Dioxide Evolution Monitoring System was also validated for measuring microbial activity in desiccated and saturated bentonite. Finally, the buffering of highly-basic bentonite at neutral pH improved the success of isolation of microbial populations, but not DNA, from the bentonite matrix. Thus, accessible techniques were optimised for exploring microbial metabolism in the atypical environments of clay matrices and desiccated conditions. These tools have application to the applied field of used nuclear fuel management, as well as for examining the fundamental biogeochemical cycles active in sedimentary and deep geological environments.