Characterization of microbial processes in deep aquifers of the Fennoscandian Shield

Micro-organisms must be included in any hydrogeochemical modelling efforts in the ongoing Swedish programme to characterize potential sites for the geological disposal of spent nuclear fuel. This paper presents the development and testing of several methods for estimating the total numbers of micro-organism groups and amounts of their biomass in groundwater, their diversity, and the rates of microbial processes. The enumeration and cultivation methods were tested and evaluated on groundwater from boreholes at 450 m depth in the Äspö Hard Rock Laboratory (HRL), Sweden, and from two potential sites for a final repository of spent nuclear fuel, Forsmark and Laxemar. The reproducibility of the methods between parallel samples and over time was investigated and found to be excellent. Nitrate-, iron-, manganese- and sulphate-reducing bacteria and acetogens and methanogens were found in numbers up to approximately 87,000 cells L⁻¹ groundwater from the studied sites. A methodology that analysed microbial process rates was developed and tested under open and closed controlled in situ conditions in a circulation system situated 447 m underground in the MICROBE laboratory at the Äspö HRL. The sulphide and acetate production rates were determined to be 0.08 and 0.14 mg L⁻¹ day⁻¹, respectively. The numbers of sulphide- and acetate-producing micro-organisms increased concomitantly in the analysed circulating groundwater. Flushing the sampled circulation aquifer created an artefact, as it lowered the sulphide concentration. Microbial and inorganic processes involved in sulphur transformations are summarized in a conceptual model, based on the observations and results presented here. The model outlines how dissolved sulphide may react with Fe(III) and Fe(II) to form solid phases of iron sulphide and pyrite. Sulphide will, consequently, continuously be removed from the aqeous phase via these reactions, at a rate approximately equalling the rate of production by microbial sulphate reduction.