Bioreduction Rate Research Articles

Effects of non-dissolved redox mediators on a hexavalent chromium bioreduction process

ABSTRACTThe effects of six non-dissolved redox mediators (RM) immobilized in cellulose acetate beads on enhancing Cr(VI) reduction by Mangrovibacter plantisponsor CR1 were investigated. In addition, the voltammetric behaviours and electron transfer capacities of the redox mediators were examined using electrochemical methods. Compared to the control beads, the Cr(VI) bioreduction rate with 1-chloroanthraquinone cellulose acetate beads (1-CAQ/CA beads) was increased up to 4.5-fold, which was mainly attributed to enhanced electron transfer by 1-CAQ. The redox mediators also improved the oxidation–reduction potential values of the Cr(VI) bioreduction processes, which might assist in Cr(VI) bioreduction. The role of the redox mediators was discussed based on the cyclic voltammetric characteristics (E0') of the redox mediators and the electrochemical impedance spectroscopy characteristics (Rct) of the RM/CA beads. A linear correlation was found for the reaction constant k and the 1-CAQ concentration (C1-CAQ), which was k = 1.5674 C1-CAQ + 4.8506 (R2 = 0.9683). The Cr(VI) bioreduction was affected by temperature, and the optimum pH for the Cr(VI) bioreduction was 6.5. The results of repeated-batch operations showed that 1-CAQ/CA beads exhibited good stability and persistence. This study contributes to a better understanding of the effects of the redox mediator on Cr(VI) bioreduction process and demonstrates its promising potential for environmental bioremediation applications.

Nitrate bioreduction in redox-variable low permeability sediments

Low permeability zone (LPZ) can play an important role as a sink or secondary source in contaminant transport in groundwater system. This study investigated the rate and end product of nitrate bioreduction in LPZ sediments. The sediments were from the U.S. Department of Energy's Hanford Site, where nitrate is a groundwater contaminant as a by-product of radionuclide waste discharges. The LPZ at the Hanford site consists of two layers with an oxidized layer on top and reduced layer below. The oxidized layer is directly in contact with the overlying contaminated aquifer, while the reduced layer is in contact with an uncontaminated aquifer below. The experimental results showed that nitrate bioreduction rate and end-product differed significantly in the sediments. The bioreduction rate in the oxidized sediment was significantly faster than that in the reduced one. A significant amount of N2O was accumulated in the reduced sediment; while in the oxidized sediment, N2O was further reduced to N2. RT-PCR analysis revealed that nosZ, the gene that codes for N2O reductase, was below detection limit in the reduced sediment. Batch experiments and kinetic modeling were performed to provide insights into the role of organic carbon bioavailability, biomass growth, and competition between nitrate and its reducing products for electrons from electron donors. The results revealed that it is important to consider sediment redox conditions and functional genes in understanding and modeling nitrate bioreduction in subsurface sediments. The results also implied that LPZ sediments can be important sink of nitrate and a potential secondary source of N2O as a nitrate bioreduction product in groundwater.

Linear free energy relationships for the biotic and abiotic reduction of nitroaromatic compounds.

Nitroaromatic compounds (NACs) are ubiquitous environmental contaminants that are susceptible to biological and abiotic reduction. Prior works have found that for the abiotic reduction of NACs, the logarithm of the NACs’ rate constants correlate with one-electron reduction potential values of the NACs (EH,NAC1) according to linear free energy relationships (LFERs). Here, we extend the application of LFERs to the bioreduction of NACs and to the abiotic reduction of NACs by bioreduced (and pasteurized) iron-bearing clay minerals. A linear correlation (R2=0.96) was found between the NACs’ bioreduction rate constants (kobs) and EH,NAC1 values. The LFER slope of log kobs versus EH,NAC1/(2.303RT/F) was close to one (0.97), which implied that the first electron transfer to the NAC was the rate-limiting step of bioreduction. LFERs were also established between NAC abiotic reduction rate constants by bioreduced iron-bearing clay minerals (montmorillonite SWy-2 and nontronite NAu-2). The second-order NAC reduction rate constants (k) by bioreduced SWy-2 and NAu-2 were well correlated to EH,NAC1 (R2=0.97 for both minerals), consistent with bioreduction results. However, the LFER slopes of log k versus EH,NAC1/(2.303RT/F) were significantly less than one (0.48–0.50) for both minerals, indicating that the first electron transfer to the NAC was not the rate-limiting step of abiotic reduction. Finally, we demonstrate that the rate of 4-acetylnitrobenzene reduction by bioreduced SWy-2 and NAu-2 correlated to the reduction potential of the clay (EH,clay, R2=0.95 for both minerals), indicating that the clay reduction potential also influences its reactivity.

Effects of aqueous uranyl speciation on the kinetics of microbial uranium reduction

The ability to predict the success of the microbial reduction of soluble U(VI) to highly insoluble U(IV) as an in situ bioremediation strategy is complicated by the wide range of geochemical conditions at contaminated sites and the strong influence of aqueous uranyl speciation on the bioavailability and toxicity of U(VI) to metal-reducing bacteria. To determine the effects of aqueous uranyl speciation on uranium bioreduction kinetics, incubations and viability assays with Shewanella putrefaciens strain 200 were conducted over a range of pH and dissolved inorganic carbon (DIC), Ca2+, and Mg2+ concentrations. A speciation-dependent kinetic model was developed to reproduce the observed time series of total dissolved uranium concentration over the range of geochemical conditions tested. The kinetic model yielded the highest rate constant for the reduction of uranyl non-carbonate species (i.e., the ‘free’ hydrated uranyl ion, uranyl hydroxides, and other minor uranyl complexes), indicating that they represent the most readily reducible fraction of U(VI) despite being the least abundant uranyl species in solution. The presence of DIC, Ca2+, and Mg2+ suppressed the formation of more bioavailable uranyl non-carbonate species and resulted in slower bioreduction rates. At high concentrations of bioavailable U(VI), however, uranium toxicity to S. putrefaciens inhibited bioreduction, and viability assays confirmed that the concentration of non-carbonate uranyl species best predicts the degree of toxicity. The effect of uranium toxicity was accounted for by incorporating the free ion activity model of metal toxicity into the bioreduction rate law. Overall, these results demonstrate that, in the absence of competing terminal electron acceptors, uranium bioreduction kinetics can be predicted over a wide range of geochemical conditions based on the bioavailability and toxicity imparted on U(VI) by solution composition. These findings also imply that the concentration of uranyl non-carbonate species, despite being extremely low, is a determining factor controlling uranium bioreduction at contaminated sites.

Cr(VI) Reduction and Cr(III) Immobilization by Acinetobacter sp. HK-1 with the Assistance of a Novel Quinone/Graphene Oxide Composite

Cr(VI) biotreatment has attracted a substantial amount of interest due to its cost effectiveness and environmental friendliness. However, the slow Cr(VI) bioreduction rate and the formed organo-Cr(III) in solution are bottlenecks for biotechnology application. In this study, a novel strain, Acinetobacter sp. HK-1, capable of reducing Cr(VI) and immobilizing Cr(III) was isolated. Under optimal conditions, the Cr(VI) reduction rate could reach 3.82 mg h(-1) g cell(-1). To improve the Cr(VI) reduction rate, two quinone/graphene oxide composites (Q-GOs) were first prepared via a one-step covalent chemical reaction. The results showed that 2-amino-3-chloro-1,4-naphthoquinone-GO (NQ-GO) exhibited a better catalytic performance in Cr(VI) reduction compared to 2-aminoanthraquinone-GO. Specifically, in the presence of 50 mg L(-1) NQ-GO, a Cr(VI) removal rate of 190 mg h(-1) g cell(-1), which was the highest rate obtained, was achieved. The increased Cr(VI) reduction rate is mainly the result of NQ-GO significantly increasing the Cr(VI) reduction activity of cell membrane proteins containing dominant Cr(VI) reductases. X-ray photoelectron spectroscopy analysis found that Cr(VI) was reduced to insoluble Cr(III), which was immobilized by glycolipids secreted by strain HK-1. These findings indicate that the application of strain HK-1 and NQ-GO is a promising strategy for enhancing the treatment of Cr(VI)-containing wastewater.

Effects of Microbial Iron Reduction and Oxidation on the Immobilization and Mobilization of Copper in Synthesized Fe(III) Minerals and Fe-Rich Soils

The effects of microbial iron reduction and oxidation on the immobilization and mobilization of copper were investigated in a high concentration of sulfate with synthesized Fe(III) minerals and red earth soils rich in amorphous Fe (hydr)oxides. Batch microcosm experiments showed that red earth soil inoculated with subsurface sediments had a faster Fe(III) bioreduction rate than pure amorphous Fe(III) minerals and resulted in quicker immobilization of Cu in the aqueous fraction. Coinciding with the decrease of aqueous Cu, SO4(2-) in the inoculated red earth soil decreased acutely after incubation. The shift in the microbial community composite in the inoculated soil was analyzed through denaturing gradient gel electrophoresis. Results revealed the potential cooperative effect of microbial Fe(III) reduction and sulfate reduction on copper immobilization. After exposure to air for 144 h, more than 50% of the immobilized Cu was remobilized from the anaerobic matrices; aqueous sulfate increased significantly. Sequential extraction analysis demonstrated that the organic matter/sulfide-bound Cu increased by 52% after anaerobic incubation relative to the abiotic treatment but decreased by 32% after oxidation, indicating the generation and oxidation of Cu-sulfide coprecipitates in the inoculated red earth soil. These findings suggest that the immobilization of copper could be enhanced by mediating microbial Fe(III) reduction with sulfate reduction under anaerobic conditions. The findings have an important implication for bioremediation in Cucontaminated and Fe-rich soils, especially in acid-mine-drainage-affected sites.

Changes in the physicochemical properties and enzymatic activity of waste during bioreduction of pig carcasses

Bioreduction is a novel method for the on-farm storage of fallen stock in a vessel containing water that is heated and aerated, prior to disposal. The combination of a mesophilic temperature and high bacterial population leads to rapid degradation of carcasses due to microbial and enzymatic breakdown of protein material; and ultimately the reduction in volume of waste to be disposed. The system could, however, be improved if more was known about the changes that occur during a bioreduction cycle. Pig carcasses were placed within two commercial-scale bioreduction vessels (BVs) (6.5 m3 capacity) and the changes in physicochemical parameters, enzymatic activity, gas emissions and microbial communities were analysed over 56 days. Analyses showed that each vessel displayed different physicochemical parameters. The microbial communities within both vessels were also distinct, though they converged between days 28 and 42 before again diverging. Of the enzymes assayed, acetylesterases showed the highest activity during initial stages, with a subsequent increase in lipase towards the end. All other enzymes showed little activity in comparison. Despite active aeration of the vessels, conditions were redox-constrained, leading to the emission of gases associated with anaerobic conditions, namely NH3 and H2S. It was concluded that no single parameter governed the biochemical processes and that each BV will have its own unique microbial population and hence rate of decomposition. Further work is needed to increase the rate of bioreduction through bioaugmentation or developing enzyme additives.

Evaluation of Fe(III)EDTA and Fe(II)EDTA-NO reduction in a NO x scrubber solution by magnetic Fe3O4-chitosan microspheres immobilized microorganisms

A two-stage bioreduction system containing magnetic-microsphere-immobilized denitrifying bacteria and iron-reducing bacteria was developed for the regeneration of scrubbing solutions for NO x removal. In this process, a higher bioreduction rate and a better tolerance of inhibition of bacteria were achieved with immobilized bacteria than with free bacteria. This work focused on evaluation of the effects of the main components in the scrubbing solution on Fe(III)EDTA (EDTA: ethylenediaminetetraacetate) and Fe(II)EDTA-NO reduction, with an emphasis on mass transfer and the kinetic model of Fe(III)EDTA and Fe(II)EDTA-NO reduction by immobilized bacteria. It was found that Fe(II)EDTA-NO had a strong inhibiting effect, but Fe(II)EDTA had no effect, on Fe(III)EDTA reduction. Fe(II)EDTA accelerated Fe(II)EDTA-NO reduction, whereas Fe(III)EDTA had no effect. This showed that the use of the two stages of regeneration was necessary. Moreover, the effect of internal diffusion on Fe(III)EDTA and Fe(II)EDTANO reduction could be neglected, and the rate-limiting step was the bioreduction process. The reduction of Fe(III)EDTA and Fe(II)EDTA-NO using immobilized bacteria was described by a first-order kinetic model. Bioreduction can therefore be enhanced by increasing the cell density in the magnetic chitosan microspheres.

Smectite Reduction by Shewanella Species as Facilitated by Cystine and Cysteine

Exogenous electron transfer mediators employed by Fe(III)-reducing bacteria are believed to govern the kinetics and equilibrium of bioreduction of Fe(III) in solid phase. In contrast to a large number of studies on humic substances and analog anthraquinone-2,6-disulfonate (AQDS), our knowledge of other potential electron shuttles involved in Fe(III) reduction is limited. The purpose of the present study was to understand the role of cystine and cysteine in reduction of iron-rich smectite (nontronite, NAu-2) by Shewanella species. A series of abiotic and biotic experiments were conducted in nongrowth media (bicarbonate buffered, pH = 7.0). Fe(II) and cysteine concentrations were monitored over the course of the bioreduction experiments with wet chemistry, and the unreduced and reduced nontronites were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results indicated that all Shewanella species tested here were capable of reducing cystine to cysteine. Either cystine or cysteine amendments significantly stimulated the Fe(III) bioreduction rate and extent. The initial reduction rate was linearly correlated with cystine or cysteine concentration. The reduction extent (18.7–22.3%) calculated from bioreactor with cystine or cysteine was slightly lower than those with AQDS (26.3%). Mineralogical analysis demonstrated that cystine or cysteine facilitated the reaction of smectite to illite as a result of Fe(III) bioreduction. Thus, we concluded that, in our experiments, cystine and cysteine functioned as electron carrier in the smectite reduction systems, and were favorable factors influencing smectite illitization.

Biotransformation of lepidocrocite in the presence of quinones and flavins

This study investigated the bioreduction of lepidocrocite (γ-FeIIIOOH) and its mineral transformation in the presence of exogenous (quinones) and endogenous (flavins) electron transfer mediators (ETMs) at low concentrations of the ETMs and bacterial cells (Shewanella putrefaciens CN32). It is very important to investigate the bioreduction of lepidocrocite in the presence of different ETMs because biotransformation of Fe(III)-containing minerals can be stimulated by ETMs and affect fate and transport of contaminants in contaminated environments. In the absence of phosphate, green rust formation was observed with fast Fe(II) production rate (0.44–0.56mMd−1) during the bioreduction of lepidocrocite with exogenous ETMs, while goethite formed at slow Fe(II) production rate (0.24–0.29mMd−1) with endogenous ETMs. In the presence of phosphate, formation of green rust and vivianite was observed with fast Fe(II) production rate (0.54–0.74mMd−1) during the bioreduction of lepidocrocite with exogenous ETMs, while vivianite formed at moderate Fe(II) production rate (0.36–0.40mMd−1) with endogenous ETMs. Vivianite formed in all experimental cases with phosphate in a broad range of Fe(II) production rates (0.23–0.74mMd−1). Our results (1) suggest that exogenous and endogenous ETMs can significantly but differently affect the biotransformation of lepidocrocite, especially at low concentrations of the ETMs and bacterial cells, (2) highlight the importance of Fe(II) production rate to determine the formation of specific biogenic minerals, (3) provide additional evidence that phosphate can significantly affect the bioreduction rate and the mineral transformation, and (4) help to understand the basic knowledge about complex interactions among microbial cell, soil mineral, and ETM in natural environments and engineered systems.

Improved cathode for high efficient microbial-catalyzed reduction in microbial electrosynthesis cells

Microbial electrosynthesis cells (MECs) are devices wherein microorganisms can electrochemically interact with electrodes, directly donating or accepting electrons from electrode surfaces. Here, we developed a novel cathode by using nickel nanowires anchored to graphite for the improvement of microbial-catalyzed reduction in MEC cathode chamber. This porous nickel-nanowire-network-coated graphite electrode increased the interfacial area and interfacial interactions between the cathode surface and the microbial biofilm. A 2.3 fold increase in bio-reduction rate over the untreated graphite was observed. Around 282 mM day(-1) m(-2) of acetate resulting from the bio-reduction of carbon dioxide by Sporomusa was produced with 82 ± 14% of the electrons consumed being recovered in acetate.

The potential impact of anaerobic microbial metabolism during the geological disposal of intermediate-level waste

AbstractMicrobial metabolism has the potential to control the biogeochemistry of redox-active radionuclides in a range of geodisposal scenarios. In this study, sediments from a high pH lime workings site were incubated under carefully controlled anaerobic conditions, at a range of alkali pH values with added electron donors and electron acceptors, to explore the limits and rates of bioreduction in a sediment system analogous to intermediate-level nuclear waste. There was a clear succession in the utilization of electron acceptors (in the order nitrate > Fe(III)-citrate > Fe(III) oxyhydroxide > sulfate), in accordance with calculated free energy yields and Eh values over the pH range 10–12. The rate and extent of bioreduction decreased at higher pH, with an upper limit for the processes studied at pH 12. The biochemical limits for such processes are discussed, alongside the potential impact of such forms of microbial metabolism on the solubility of a range of redox active radionuclides that will feature heavily in the safety case for the geological disposal of intermediate-level nuclear waste.

Enhanced Reduction of Fe(II)EDTA-NO/Fe(III)EDTA in NOx Scrubber Solution Using a Three-Dimensional Biofilm-Electrode Reactor

A promising technique called chemical absorption-biological reduction (CABR) integrated approach has been developed recently for the nitrogen oxides (NO(x)) removal from flue gases. The major challenge for this approach is how to enhance the rate of the biological reduction step. To tackle the challenge, a three-dimensional biofilm-electrode reactor (3D-BER) was utilized. This reactor provides not only considerable amount of sites for biofilm, but also many electron donors for bioreduction. Factors affecting the performance of 3D-BER were optimized, including material of the third electrode (graphite), glucose concentration (1000 mg·L(-1)), and volume current density (30.53 A·m(-3) NCC). Experimental results clearly demonstrated that this method significantly promotes the bioreduction rate of Fe(II)EDTA-NO (0.313 mmol·L(-1)·h(-1)) and Fe(III)EDTA (0.564 mmol·L(-1)·h(-1)) simultaneously. Experiments on the mechanism showed that Fe(II)EDTA serves as the primary electron donor in the reduction of Fe(II)EDTA-NO, whereas the reduction of Fe(III)EDTA took advantage of both glucose and electrolysis-generated H(2) as electron donors. High concentration of Fe(II)EDTA-NO or Fe(III)EDTA interferes the bioreduction of the other one. The proposed methodology shows a promising prospect for NO(x) removal from flue gas.

Hexavalent Chromium Bioremoval through Adaptation and Consortia Development from Sukinda Chromite Mine Isolates

Indigenous isolates from the waters of chromite mining sites at Sukinda Valley, Orissa, India, showed a considerable enhancement in Cr (VI) bioreduction rate through adaptation and consortia develo...

A biogeochemical framework for bioremediation of plutonium(V) in the subsurface environment

Accidental release of plutonium (Pu) from storage facilities in the subsurface environment is a concern for the safety of human beings and the environment. Given the complexity of the subsurface environment and multivalent state of Pu, we developed a quantitative biogeochemical framework for bioremediation of Pu(V)O(2) (+) in the subsurface environment. We implemented the framework in the biogeochemical model CCBATCH by expanding its chemical equilibrium for aqueous complexation of Pu and its biological sub-models for including Pu's toxicity and reduction reactions. The quantified framework reveals that most of the Pu(V) is speciated as free Pu(V)O(2) (+) ((aq)), which is a problem if the concentration of free Pu(V)O(2) (+) is ≥28μM (the half-maximum toxicity value for bacteria able to reduce Pu(V) to Pu(III)PO(4(am))) or ≥250μM (the full-toxicity value that takes the bioreduction rate to zero). The framework includes bioreduction of Fe(3+) to Fe(2+), which abiotically reduces Pu(V)O(2) (+) to Pu(IV) and then to Pu(III). Biotic (enzymatic) reduction of Pu(V)O(2) (+) directly to Pu(III) by Shewanella alga (S. alga) is also included in the framework. Modeling results also reveal that for formation of Pu(III)PO(4(am)), the desired immobile product, the concentration of coexisting model strong ligand-nitrilotriacetic acid (NTA)-should be less than or equal to the concentration of total Pu(III).

Microbial reduction of Fe(III) in illite–smectite minerals by methanogen Methanosarcina mazei

Clay minerals are ubiquitous in soils, sediments and sedimentary rocks, and could coexist with methanogens in anoxic environments, but whether or not methanogens can reduce structural Fe(III) in clay minerals other than smectite is not known. The objective of this study was to understand the capability of a mesophilic methanogen, Methanosarcina mazei in reducing structural Fe(III) in illite–smectite clay minerals and its potential role in inducing mineralogical changes. Bioreduction experiments were performed in batch systems, where four different clay minerals (nontronite NAu-2, mixed-layer illite–smectite RAr-1 and ISCz-1, and illite IMt-1) were exposed to M. mazei in growth medium with and without anthraquinone-2,6-disulfonate (AQDS) as electron shuttle. The extent and rate of bioreduction were quantified via wet chemistry and mineralogical transformed by X-ray diffraction and scanning and transmission electron microscopy. Our results demonstrated that M. mazei was able to reduce structural Fe(III) in clay minerals with Fe(III) in smectite being the most reducible and illite the least. AQDS enhanced the reduction rate and extent. The bioavailability of structural Fe(III) to M. mazei was correlated to the smectite proportion in each clay mineral . Methanogenesis was inhibited by Fe(III) bioreduction, possibly due to diversion of electrons from the methanogenesis pathway to structural Fe(III) in clay minerals. Bioreduction of Fe(III) in clay minerals induced formation of biogenic mixed-layer illite–smectite, silica, and vivianite. These data collectively showed that mesophilic methanogen is capable of reducing structural Fe(III) in illite–smectite minerals and of inducing a number of mineralogical changes.

Effects of redox cycling of iron in nontronite on reduction of technetium

In situ technetium-99 ( 99Tc) immobilization by Fe(II) associated with clay minerals is a potential cost-effective method for Tc remediation at contaminated sites. The oxidation state of Fe is constantly cycled in sedimentary environments, however the effect of this cycle on Tc reduction and immobilization has not yet been investigated. The objective of this project was therefore to study how multiple cycles of reduction–reoxidation of Fe-rich clay mineral, nontronite, affected its reactivity toward Tc (VII) reduction. Iron-rich nontronite NAu-2 was used as a model clay mineral. NAu-2 suspension was first bioreduced by Shewanella putrefaciens CN32, which was subsequently re-oxidized by air. Three cycles of reduction–oxidation were conducted and bioreduced NAu-2 samples from all three cycles were collected and used for Tc(VII) reduction experiments. Each redox cycle resulted in a small fraction of dissolution of small size and/or poorly crystalline NAu-2. The released Fe(II) from the dissolution was likely adsorbed onto NAu-2 surface/edge sites with a high reactivity. Upon exposure to O 2, this reactive Fe(II) fraction was oxidized more rapidly than structural Fe(II) and may have accounted for a two-step reoxidation kinetics of NAu-2 associated Fe(II): rapid oxidation over first few hours followed by slow oxidation. Progressive increase of this reactive fraction of Fe(II), from increased dissolution, accounted for the successively higher rate of bioreduction and reoxidation with increased redox cycles. The same Fe redistribution accounted for two-step Tc(VII) reduction kinetics as well. Rapid Tc(VII) reduction in the first few hours may be attributed to a small fraction of highly reactive Fe(II) at the NAu-2 surface/edge sites, and more steady Tc(VII) reduction over longer time may be carried out by structural Fe(II). Similar to the increased rates of Fe(III) reduction and Fe(II) oxidation, the Tc(VII) reduction rate also increased with redox cycles and could be explained by progressive increase of the reactive Fe(II) on NAu-2 surface/edges. Iron-rich clay minerals undergo important changes after redox cycles, but eventually reach a steady state with continued reactivity toward heavy metals.

Effects of Si-bearing minerals on the nature of secondary iron mineral products from lepidocrocite bioreduction

Lepidocrocite (γ-FeOOH) bioreduction by Shewanella putrefaciens was examined in systems containing three kinds of quartz silica sand with solid contents ranging from 5 to 70 wt.%. The results notably demonstrated that the formation routes of secondary iron minerals from microbial reduction of γ-FeOOH in silica mineral assemblages are dependent on quartz sand loading and particle size distribution. Biomineralization experiments with a concentration range of aqueous silica showed that a lower Si concentration (less than 2 mM) led to magnetite or magnetite/chukanovite, while higher concentrations resulted in the formation of green rust (GR). Dissolution of very fine quartz particles may have promoted the formation of biogenic GR instead of magnetite. This behavior could be explained by the sorption of dissolved silicate on the lepidocrocite surface, which might limit the bioreduction rate and promote GR formation. As observed with the chemically synthesized GR, the interactions of dissolved or colloidal Si with the lateral faces of the GR crystals could stabilize the GR structure, thus preventing its transformation into other products. Furthermore, complex physical and chemical distribution at the surfaces and within heteroaggregates of mixed mineral phases may affect the diffusion processes of species and therefore the biomineralization mechanism.

Bioreduction of Fe-bearing clay minerals and their reactivity toward pertechnetate (Tc-99)

99Technetium ( 99Tc) is a fission product of uranium-235 and plutonium-239 and poses a high environmental hazard due to its long half-life ( t 1/2 = 2.13 × 10 5 y), abundance in nuclear wastes, and environmental mobility under oxidizing conditions [i.e., Tc(VII)]. Under reducing conditions, Tc(VII) can be reduced to insoluble Tc(IV). Ferrous iron, either in aqueous form (Fe 2+) or in mineral form [Fe(II)], has been used to reduce Tc(VII) to Tc(IV). However, the reactivity of Fe(II) from clay minerals, other than nontronite, toward immobilization of Tc(VII) and its role in retention of reduced Tc(IV) has not been investigated. In this study the reactivity of a suite of clay minerals toward Tc(VII) reduction and immobilization was evaluated. The clay minerals chosen for this study included five members in the smectite–illite (S–I) series, (montmorillonite, nontronite, rectorite, mixed layered I–S, and illite), chlorite, and palygorskite. Surface Fe-oxides were removed from these minerals with a modified dithionite–citrate–bicarbonate (DCB) procedure. The total structural Fe content of these clay minerals, after surface Fe-oxide removal, ranged from 0.7% to 30.4% by weight, and the structural Fe(III)/Fe(total) ratio ranged from 45% to 98%. X-ray diffraction (XRD) and Mössbauer spectroscopy results showed that after Fe oxide removal the clay minerals were free of Fe-oxides. Scanning electron microscopy (SEM) revealed that little dissolution occurred during the DCB treatment. Bioreduction experiments were performed in bicarbonate buffer (pH-7) with structural Fe(III) in the clay minerals as the sole electron acceptor, lactate as the sole electron donor, and Shewanella putrefaciens CN32 cells as a mediator. In select tubes, anthraquinone-2,6-disulfate (AQDS) was added as electron shuttle to facilitate electron transfer. In the S–I series, smectite (montmorillonite) was the most reducible (18% and 41% without and with AQDS, respectively) and illite the least (1% for both without and with AQDS). The extent and initial rate of bioreduction were positively correlated with the percent smectite in the S–I series (i.e., layer expandability). Fe(II) in the bioreduced clay minerals subsequently was used to reduce Tc(VII) to Tc(IV) in PIPES buffer. Similar to the trend of bioreduction, in the S–I series, reduced NAu-2 showed the highest reactivity toward Tc(VII), and reduced illite exhibited the least. The initial rate of Tc(VII) reduction, after normalization to clay and Fe(II) concentrations, was positively correlated with the percent smectite in the S–I series. Fe(II) in chlorite and palygorskite was also reactive toward Tc(VII) reduction. These data demonstrate that crystal chemical parameters (layer expandability, Fe and Fe(II) contents, and surface area, etc.) play important roles in controlling the extent and rate of bioreduction and the reactivity toward Tc(VII) reduction. Reduced Tc(IV) resides within clay mineral matrix, and this association could minimize any potential of reoxidation over long term.

How does organic matter constrain the nature, size and availability of Fe nanoparticles for biological reduction?

Few studies have so far examined the kinetics and extent of the formation of Fe-colloids in the presence of natural organic ligands. The present study used an experimental approach to investigate the rate and amount of colloidal Fe formed in presence of humic substances, by gradually oxidizing Fe(II) at pH 6.5 with or without humic substances (HS) (in this case, humic acid – HA and fulvic acid – FA). Without HS, micronic aggregates (0.1–1μm diameter) of nano-lepidocrocite is obtained, whereas, in a humic-rich medium (HA and FA suspensions at 60 and 55ppm of DOC respectively), nanometer-sized Fe particles are formed trapped in an organic matrix. A proportion of iron is not found to contribute to the formation of nanoparticles since iron is complexed to HS as Fe(II) or Fe(III). Humic substances tend to (i) decrease the Fe oxidation and hydrolysis, and (ii) promote nanometer-sized Fe oxide formation by both inhibiting the development of hydroxide nuclei and reducing the aggregation of Fe nanoparticles.Bioreduction experiments demonstrate that bacteria (Shewanella putrefaciens CIP 80.40T) are able to use Fe nanoparticles associated with organic matter about eight times faster than in the case of nano-lepidocrocite. This increase in bioreduction rate appears to be related to the presence of humic acids that (i) indirectly control the size, shape and density of oxyhydroxides and (ii) directly enhance biological reduction of nanoparticles by electron shuttling and Fe complexation. These results suggest that, in wetlands but also elsewhere where mixed organic matter-Fe colloids occur, Fe nanoparticles closely associated with organic matter represent a bioavailable Fe source much more accessible for microfauna than do crystallized Fe oxyhydroxides.