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The formation of pedogenic carbonate (calcrete) in terrestrial environments is commonly mediated by microorganisms. In Australia, Au-anomalous calcrete is an important sampling medium for geochemical exploration, but current models describing its formation do not include a confirmed microbial component. This study demonstrates that bacterial communities in calcareous sands from dunes overlying the Barns Cold Deposit in semi-arid South Australia, are capable of mediating the biomineralisation of Au-anomalous carbonates. Bacterial enrichment cultures obtained from calcareous sands at three depths (0.1, 0.64 and 2.1 m, plus abiotic control) were incubated in urea and Ca(2+)-containing growth media (pH 8), unamended and amended with Au (100 parts-per-billion, ppb) as Au-aspartic-acid complex. During the incubation of the enrichment cultures urea was turned over to NH(4)(+) within 96 h to 220 h. The solution pH increased concurrently by approximately 1.2 units, and Au-anomalous Ca-carbonate crystallites were precipitated on cells, which functioned as nucleation sites: no carbonate precipitation was observed in abiotic controls. Compared to the medium, Au was strongly enriched in these carbonates and appeared to be uniformly dispersed in the individual crystallites, as shown using LA-ICP-MS; a similar distribution is present in naturally occurring Auanomalous calcrete. Phylogenetic 16S rRNA PCR DGGE analyses, shotgun cloning and functional microbial analyses (BioLog, ureC quantitative PCR) demonstrated that naturally occurring and culture-enriched bacterial communities were dominated by alkaliphylic, halotolerant Bacillus spp. The indigenous bacterial communities were capable of utilising amino acids (including L-aspartic acid) and urea, which appears to lead to the destabilisation of the Au-amino acid complexes and concomitant co-precipitation of Au in the Ca-carbonates. In conclusion, a model combining geomicrobial- with evapotranspiration- and plant-based components is likely to best describe the formation of (Au-anomalous) calcrete in semi-arid and and zones. Crown Copyright (C) 2008 Published by Elsevier B.V. All rights reserved.