The in situ stimulation of Fe(III) oxide reduction by Geobacter bacteria leads to the concomitant precipitation of hexavalent uranium [U(VI)] from groundwater. Despite its promise for the bioremediation of uranium contaminants, the biological mechanism behind this reaction remains elusive. Because Fe(III) oxide reduction requires the expression of Geobacter 's conductive pili, we evaluated their contribution to uranium reduction in Geobacter sulfurreducens grown under pili-inducing or noninducing conditions. A pilin-deficient mutant and a genetically complemented strain with reduced outer membrane c -cytochrome content were used as controls. Pili expression significantly enhanced the rate and extent of uranium immobilization per cell and prevented periplasmic mineralization. As a result, pili expression also preserved the vital respiratory activities of the cell envelope and the cell's viability. Uranium preferentially precipitated along the pili and, to a lesser extent, on outer membrane redox-active foci. In contrast, the pilus-defective strains had different degrees of periplasmic mineralization matching well with their outer membrane c -cytochrome content. X-ray absorption spectroscopy analyses demonstrated the extracellular reduction of U(VI) by the pili to mononuclear tetravalent uranium U(IV) complexed by carbon-containing ligands, consistent with a biological reduction. In contrast, the U(IV) in the pilin-deficient mutant cells also required an additional phosphorous ligand, in agreement with the predominantly periplasmic mineralization of uranium observed in this strain. These findings demonstrate a previously unrecognized role for Geobacter conductive pili in the extracellular reduction of uranium, and highlight its essential function as a catalytic and protective cellular mechanism that is of interest for the bioremediation of uranium-contaminated groundwater.

Biofilms formed by dissimilatory metal reducers are of interest to develop permeable biobarriers for the immobilization of soluble contaminants such as uranium. Here we show that biofilms of the model uranium-reducing bacterium Geobacter sulfurreducens immobilized substantially more U(VI) than planktonic cells and did so for longer periods of time, reductively precipitating it to a mononuclear U(IV) phase involving carbon ligands. The biofilms also tolerated high and otherwise toxic concentrations (up to 5 mM) of uranium, consistent with a respiratory strategy that also protected the cells from uranium toxicity. The enhanced ability of the biofilms to immobilize uranium correlated only partially with the biofilm biomass and thickness and depended greatly on the area of the biofilm exposed to the soluble contaminant. In contrast, uranium reduction depended on the expression of Geobacter conductive pili and, to a lesser extent, on the presence of the c cytochrome OmcZ in the biofilm matrix. The results support a model in which the electroactive biofilm matrix immobilizes and reduces the uranium in the top stratum. This mechanism prevents the permeation and mineralization of uranium in the cell envelope, thereby preserving essential cellular functions and enhancing the catalytic capacity of Geobacter cells to reduce uranium. Hence, the biofilms provide cells with a physically and chemically protected environment for the sustained immobilization and reduction of uranium that is of interest for the development of improved strategies for the in situ bioremediation of environments impacted by uranium contamination.

Quantitative microbial risk assessment (QMRA) is a powerful decision analytics tool, yet it faces challenges when modeling health risks for the indoor environment. One limitation is uncertainty in fomite recovery for evaluating the efficiency of decontamination. Addressing this data gap has become more important as a result of response and recovery from a potential malicious pathogen release. To develop more accurate QMRA models, recovery efficiency from non-porous fomites (aluminum, ceramic, glass, plastic, steel, and wood laminate) was investigated. Fomite material, surface area (10, 100, and 900 cm(2)), recovery tool (swabs and wipes), initial concentration on the fomites and eluent (polysorbate 80, trypticase soy broth, and beef extract) were evaluated in this research. Recovery was shown to be optimized using polysorbate 80, sampling with wipes, and sampling a surface area of 10-100 cm(2). The QMRA model demonstrated, through a relative risk comparison, the need for recovery efficiency to be used in these models to prevent underestimated risks.

Although the ability of some microorganisms to grow without O2 has long been recognized, the application of new methodologies has greatly expanded the known diversity and potential of anaerobic microorganisms and processes. In particular, anaerobic techniques that permit the successful cultivation of microorganisms on solid media have opened new avenues for the study of the physiology and metabolic potential of many new microorganisms using molecular, genomic, and proteomic tools. One technique above all has proven instrumental for anaerobic studies over the years: the use of the anaerobic chamber. This unit gives a brief description of the methods used for the cultivation of anaerobic microorganisms, and describes in detail the principles and applications of anaerobic chambers, with special emphasis on vinyl glove boxes. The methodologies described in this unit should provide the interested but inexperienced investigator with the basic tools to successfully cultivate anaerobic microorganisms and study anaerobic processes. Curr. Protoc. Microbiol. 12:A.4F.1‐A.4F.16. © 2009 by John Wiley & Sons, Inc.

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