Currently, the production of radioactive waste from nuclear industries is increasing, leading to the development of reliable containment strategies. The deep geological repository (DGR) concept has emerged as a suitable storage solution, involving the underground emplacement of nuclear waste within stable geological formations. Bentonite clay, known for its exceptional properties, serves as a critical artificial barrier in the DGR system. Recent studies have suggested the stability of bentonite within DGR relevant conditions, indicating its potential to enhance the long-term safety performance of the repository. On the other hand, due to its high resistance to corrosion, copper is one of the most studied reference materials for canisters. This review provides a comprehensive perspective on the influence of nuclear waste conditions on the characteristics and properties of DGR engineered barriers. This paper outlines how evolving physico-chemical parameters (e.g., temperature, radiation) in a nuclear repository may impact these barriers over the lifespan of a repository and emphasizes the significance of understanding the impact of microbial processes, especially in the event of radionuclide leakage (e.g., U, Se) or canister corrosion. Therefore, this review aims to address the long-term safety of future DGRs, which is critical given the complexity of such future systems.

Deep geological repositories (DGRs) stand out as one of the optimal options for managing high-level radioactive waste (HLW) such as uranium (U) in the near future. Here, we provide novel insights into microbial behavior in the DGR bentonite barrier, addressing potential worst-case scenarios such as waste leakage (e.g., U) and groundwater infiltration of electron rich donors in the bentonite. After a three-year anaerobic incubation, Illumina sequencing results revealed a bacterial diversity dominated by anaerobic and spore-forming microorganisms mainly from the phylum Firmicutes. Highly U tolerant and viable bacterial isolates from the genera Peribacillus, Bacillus, and some SRB such as Desulfovibrio and Desulfosporosinus, were enriched from U-amended bentonite. The results obtained by XPS and XRD showed that U was present as U(VI) and as U(IV) species. Regarding U(VI), we have identified biogenic U(VI) phosphates, U(UO₂)·(PO₄)_2, located in the inner part of the bacterial cell membranes in addition to U(VI)-adsorbed to clays such as montmorillonite. Biogenic U(IV) species as uraninite may be produced as result of bacterial enzymatic U(VI) reduction. These findings suggest that under electron donor-rich water-saturation conditions, bentonite microbial community can control U speciation, immobilizing it, and thus enhancing future DGR safety if container rupture and waste leakage occurs.

Uranium (U) represents the preeminent hazardous radionuclide within the context of nuclear waste repositories. Indigenous microorganisms in bentonite can influence radionuclide speciation and migration in Deep Geological Repositories (DGRs) for nuclear waste storage. While bacterial communities in bentonite samples have been extensively studied, the impact of fungi has been somewhat overlooked. Here, we investigate the geomicrobiological processes in bentonite microcosms amended with uranyl nitrate and glycerol-2-phosphate (G2P) for six-month incubation. ITS sequencing revealed that the fungal community was mainly composed of Ascomycota (96.6 %). The presence of U in microcosms enriched specific fungal taxa, such as Penicillium and Fusarium, potentially associated with uranium immobilization mechanisms. Conversely, the amendment of U into G2P-suplemented samples exhibited minimal impact, resulting in a fungal community akin to the control group. Several fungal strains were isolated from bentonite microcosms to explore their potential in the U biomineralization, including Fusarium oxysporum, Aspergillus sp., Penicillium spp., among others. High Annular Angle Dark-Field Scanning Transmission Electron Microscopy (HAADF) analyses showed the capacity of F. oxysporum B1 to form U-phosphate mineral phases, likely mediated by phosphatase activity. Therefore, our study emphasizes the need to take into account indigenous bentonite fungi in the overall assessment of the impact of microbial processes in the immobilization of U within DGRs environments.

The deep geological repository (DGR) concept consists of storing radioactive waste in metal canisters, surrounded by compacted bentonite, and placed deeply into a geological formation. Here, bentonite slurry microcosms with copper canisters, inoculated with bacterial consortium and amended with acetate, lactate and sulfate were set up to investigate their geochemical evolution over a year under anoxic conditions. The impact of microbial communities on the corrosion of the copper canisters in an early-stage (45 days) was also assessed. The amended bacterial consortium and electron donors/acceptor accelerated the microbial activity, while the heat-shocked process had a retarding effect. The microbial communities partially oxidize lactate to acetate, which is subsequently consumed when the lactate is depleted. Early-stage microbial communities showed that the bacterial consortium reduced microbial diversity with Pseudomonas and Stenotrophomonas dominating the community. However, sulfate-reducing bacteria such as Desulfocurvibacter, Anaerosolibacter, and Desulfosporosinus were enriched coupling oxidation of lactate/acetate with reduction of sulfates. The generated biogenic sulfides, which could mediate the conversion of copper oxides (possibly formed by trapped oxygen molecules on the bentonite or driven by the reduction of H₂O) to copper sulfide (Cu₂S), were identified by X-ray photoelectron spectroscopy (XPS). Overall, these findings shed light on the ideal geochemical conditions that would affect the stability of DGR barriers, emphasizing the impact of the SRB on the corrosion of the metal canisters, the gas generation, and the interaction with components of the bentonite.

One of the major challenges for future Deep Geological Repositories (DGRs) for radioactive waste is to ensure long-term safety over timescales exceeding 100,000 years. The effect of gamma radiation on bentonite microorganisms remains poorly understood, with even less research exploring its combined impact with other key repository conditions. This study investigates the combined effects of high bentonite compaction density (1.6 g/cm³) and gamma radiation (cumulative doses 14 kGy or 28 kGy) on FEBEX bentonite blocks under complete water saturation with synthetic pore water, anoxic atmosphere, and sulfate-reducing bacteria (SRB). Culture-dependent microbiological analyses and next-generation sequencing demonstrated that native FEBEX bentonite microorganisms can withstand these challenging conditions, favoring spore-forming bacteria. After 1-year incubation, in both non-irradiated and irradiated samples, extremophile genera like Saccharopolyspora and Streptomyces, were identified. In addition, gamma radiation negatively impacted heterotrophic aerobe viability. However, a key finding was that a 6-month incubation period prior to radiation exposure enhanced microbial resistance to this stressor. Mineralogically, neither gamma radiation nor other studied conditions affected the properties of FEBEX bentonite, which remained stable. This study provides groundbreaking insights into the combined effects of gamma radiation and other DGR-relevant conditions on bentonite microbiology and mineralogy, significantly advancing the field.