Toxic metals can be immobilized on surface sorption sites of soil Fe(III) minerals or can be included in the mineral structure (4, 29). Fe(III)-reducing bacteria (FeRB) can facilitate the release of these metals by reductive dissolution of Fe(III) oxides (9, 17) and bioreduction of Fe(III) oxide-bound trace metals (42). This release might enhance metal stress, suggesting that metal tolerance should be an important attribute for FeRB. Acidophilic FeRB can tolerate millimolar concentrations of Cd, Cu, Ni, and Zn (12), which might be a prerequisite to survival in habitats where low pH facilitates high metal solubility. In contrast, neutrophilic FeRB like Shewanella spp. tolerate only M concentrations (34, 36). Geobacter spp. have not been tested to the best of our knowledge, but metal tolerance proteins are expressed during growth in uranium-contaminated sediments, which might be connected to metal resistance (19).Near Ronneburg (Thuringia, Germany), uranium mining caused severe environmental contamination with metals and radionuclides (20). In creek bank alluvial soils of the Gessenbach, a main drainage system of upstream mining sites (41), high heavy metal concentrations occur both in solid phase and in the pore water of a ground-and surface water-influenced, oxidized, iron-rich Btlc horizon of a Luvic Gleysol. We demonstrated the solubilization of Co, Ni, Zn, As, and U in Btlc soil microcosms during biostimulated microbial Fe(III) reduction that was associated with the activity of microorganisms related to Delta-and Betaproteobacteria, Acidobacteria, and Firmicutes (7). The aims of this study were to (i) determine the heavy metal fraction of the solid phase, which could be released during reductive dissolution of Fe(III) oxides, (ii) estimate the effect of heavy metals on the activity of FeRB in the Gessenbach creek bank soil, and (iii) identify metal-tolerant FeRB, because the permanent exposure to contaminants during the last 50 years should have promoted metal tolerance.Soil geochemistry. Putative binding forms of heavy metals in the Btlc soil solid phase were determined by sequential extraction (8, 43) in samples collected in August 2006. Metal concentrations were analyzed with either ICP-MS (inductively coupled plasma-mass spectrometry) or ICP-OES (optical emission spectrometry) (8). Most metals (20 to 40%) and even 80% of As in Btlc soil were detected in fraction 5, which is representative for amorphous Fe(III) oxides (Fig. 1). A considerable amount of uranium (30%) was recovered in the specifically adsorbed fraction, whereas only Zn and Ni were primarily recovered in the mobile fraction. Zn and Ni also dominated the heavy metal pore water concentration of the creek bank soil, which was sampled monthly from June to November 2007 (7). Pore water heavy metal and As concentrations always peaked in the Btlc horizon (see Fig. S1 in the supplemental material) and reached maximum concentrations of 38.6, 16.4, 3.9, 1.5, 0.6, and 0.3 M for Zn, Ni, Al, Cu, Co, and Cd, respectively. Pore water Fe(II) concentrations, ...