Microbiology of barrier component analogues of a deep geological repository

Abstract
Canada is currently implementing a site selection process to identify a location for a deep geological repository (DGR) for the long-term storage of Canada’s used nuclear fuel, wherein used nuclear fuel bundles will be sealed inside copper-coated carbon steel containers, encased in highly compacted bentonite clay buffer boxes and sealed deep underground in a stable geosphere. Because a DGR must remain functional for a million years, there is value to examining ancient natural systems that serve as analogues for planned DGR components. Specifically, studying the microbiology of natural analogue components of a DGR is important for developing an understanding of the types of microorganisms that may be able to grow and influence the long-term stability of a DGR. This study explored the abundance, viability, and composition of microorganisms in several ancient natural analogues using a combination of cultivation and cultivation-independent approaches. Samples were obtained from the Tsukinuno bentonite deposit (Japan) that formed ~10 mya, the Opalinus Clay formation (Switzerland) that formed ~174 mya, and Canadian shield crystalline rock from Northern Ontario that formed ~2.7 bya. Analysis of 16S rRNA gene amplicons revealed that three of the ten Tsukinuno bentonite samples analyzed were dominated by putative aerobic heterotrophs and fermenting bacteria from the Actinobacteria phylum, whereas five of the Tsukinuno bentonite samples were dominated by sequences associated with putative acidophilic chemolithoautotrophs capable of sulfur reduction.