Nontronite NAu-2 Research Articles

Critical Role of Mineral Fe(IV) Formation in Low Hydroxyl Radical Yields during Fe(II)-Bearing Clay Mineral Oxygenation.

In subsurface environments, Fe(II)-bearing clay minerals can serve as crucial electron sources for O2 activation, leading to the sequential production of O2•-, H2O2, and •OH. However, the observed •OH yields are notably low, and the underlying mechanism remains unclear. In this study, we investigated the production of oxidants from oxygenation of reduced Fe-rich nontronite NAu-2 and Fe-poor montmorillonite SWy-3. Our results indicated that the •OH yields are dependent on mineral Fe(II) species, with edge-surface Fe(II) exhibiting significantly lower •OH yields compared to those of interior Fe(II). Evidence from in situ Raman and Mössbauer spectra and chemical probe experiments substantiated the formation of structural Fe(IV). Modeling results elucidate that the pathways of Fe(IV) and •OH formation respectively consume 85.9-97.0 and 14.1-3.0% of electrons for H2O2 decomposition during oxygenation, with the Fe(II)edge/Fe(II)total ratio varying from 10 to 90%. Consequently, these findings provide novel insights into the low •OH yields of different Fe(II)-bearing clay minerals. Since Fe(IV) can selectively degrade contaminants (e.g., phenol), the generation of mineral Fe(IV) and •OH should be taken into consideration carefully when assessing the natural attenuation of contaminants in redox-fluctuating environments.

Mechanistic Insight into Electron Transfer from Fe(II)-Bearing Clay Minerals to Fe (Hydr)oxides.

Electron transfer (ET) is the essence of most biogeochemical processes related to element cycling and contaminant attenuation, whereas ET between different minerals and the controlling mechanism remain elusive. Here, we used surface-associated Fe(II) as a proxy to explore ET between reduced nontronite NAu-2 (rNAu-2) and Fe (hydr)oxides in their coexisting systems. Results showed that ET could occur from rNAu-2 to ferrihydrite but not to goethite, and the ET amount was determined by the number of reactive sites and the reduction potential difference between rNAu-2 and ferrihydrite. ET proceeded mainly through the mineral-mineral interface, with a negligible contribution of dissolved Fe2+/Fe3+. Control experiments by adding K+ and increasing salinity together with characterizations by X-ray diffraction, scanning electron microscopy/energy-dispersive spectrometry, and atomic force microscopy suggested that ferrihydrite nanoparticles inserted the interlayer space in rNAu-2 where structural Fe(II) in rNAu-2 transferred electrons mainly through the basal plane to ferrihydrite. This study implicates the occurrence of ET between different redox-active minerals through the mineral-mineral interface. As minerals at different reduction potentials often coexist in soils/sediments, the mineral-mineral ET may play an important role in subsurface biogeochemical processes.

Microbial reduction and alteration of Fe(III)-containing smectites in the presence of biochar-derived dissolved organic matter

Clay minerals are important components of solid-phase iron pools in soils, and the redox state of structurally-coordinated iron in clay minerals can significantly influence soil physicochemical properties. Due to increasing use of biochar in agriculture, the dissolved organic matter derived from biochar (BDOM) has become an unignorable fraction of dissolved organic carbon in soil systems. However, our knowledge regarding the effect of BDOM on soil biogeochemical processes (particularly on iron redox cycling) is still very limited. To examine the role of BDOM in the microbial reduction of Fe(III) in clays, BDOM was extracted from wheat straw biochar and selectively added to laboratory reactors in which lactate, Fe(III)-containing smectite (montmorillonite SWy-2 or nontronite NAu-2), and a typical iron-reducing bacterium Shewanella oneidensis strain MR-1 were used as the respective electron donor, electron acceptor, and reaction mediator. The results demonstrated that BDOM contained bio-reactive quinone moieties. BDOM amendment increased the initial rate and final extent of Fe(III) bioreduction by strain MR-1. Such promotion effect of BDOM might be ascribed to its quinone moieties functioning as electron shuttles between MR-1 cells and structural Fe(III) in smectite. Structural changes were observed in the bioreduced smectite systems, especially for NAu-2 reactors in which obvious dissolution and illitization occurred. These observations suggest that BDOM is bio-reactive and is able to stimulate the microbial reduction of Fe(III) in clay minerals, and may offer new insights into the effects of biochar application in elemental cycles in soils.

Effect of Structural Fe Reduction on Water Sorption by Swelling and Non-Swelling Clay Minerals

Ferruginous clay minerals in saturated soils and within hydrocarbon deposits often exist in a reduced state. Upon introduction of dissolved oxygen, or other oxidants, the clay minerals oxidise and changes in mineral surface charge and sorption capacity occur, resulting in changes in hydration as well as flux of intercalated species. Here we examine the sorption of water to the Fe-containing clay minerals nontronite NAu-2 (23 wt% Fe) and illite IMt-2 (7 wt% Fe) as a function of Fe oxidation state and exchangeable cations by means of water vapour volumetry and N2 surface area analysis. The clay minerals were chemically reduced using sodium dithionite. Sorption isotherms of water vapour and nitrogen, controlled relative humidity diffractograms, and chemical analyses were recorded. The results show that, after reduction using sodium dithionite, increased amounts of water vapour and nitrogen were adsorbed to the high Fe content nontronite, despite decreased interlayer separation. Little change was observed for the non-swelling and low Fe content illite. Sodium from the reducing agent was found to exchange with calcium present in the starting clay minerals, and sodium balanced the additional mineral charge generated during reduction. The findings presented in this study deliver improved understanding of sorption at the surface of the reduced clay minerals, which aid constrain the role of clay mineral interfaces in subsurface environments.

Sustainable enhancement of Cr(VI) bioreduction by the isolated Cr(VI)-resistant bacteria

Bioreduction of mobile Cr(VI) to sparingly soluble Cr(III) is an effective strategy for in situ remediations of Cr contaminated sites. The key of this technology is to screen Cr(VI)-resistant bacteria and further explore the sustainable enhancement approaches towards their Cr(VI) reduction performance. In this study, a total of ten Cr(VI)-resistant bacteria were isolated from a Cr(VI) contaminated site. All of them could reduce Cr(VI), and the greatest extent of Cr(VI) reduction (98%) was obtained by the isolated CRB6 strain. The isolated CRB6 was able to reduce structural Fe(III) in Nontronite NAu-2 to structural Fe(II). Compared with the slow bioreduction process, the produced structural Fe(II) can rapidly enhance Cr(VI) reduction. The resist dissolution characteristics of NAu-2 in the redox cycling may provide sustainable enhancement of Cr(VI) reduction. However, no enhancement on Cr(VI) bioreduction by the isolated CRB6 was observed in the presence of NAu-2, which was attributed to the inhibition of Cr(VI) on the electron transfer between the isolated CRB6 and NAu-2. AQDS can accelerate the electron transfer between the isolated CRB6 and NAu-2 as an electron shuttle in the presence of Cr(VI). Therefore, the combination of NAu-2 and AQDS generated a synergistic enhancement on Cr(VI) bioreduction compared with the enhancement obtained by NAu-2 and AQDS individually. Our results highlight that structural Fe(III) and electron shuttle can provide a sustainable enhancement of Cr(VI) reduction by Cr(VI)-reducing bacteria, which has great potential for the effective Cr(VI) in-situ remediation.

Antibacterial Mechanisms of Reduced Iron-Containing Smectite-Illite Clay Minerals.

Reduced nontronite has been demonstrated to be antibacterial through the production of hydroxyl radical (•OH) from the oxidation of structural Fe(II). Herein, we investigated the antibacterial activity of more common smectite-illite (S-I) clays toward Escherichia coli cells, including montmorillonite SWy-3, illite IMt-2, 50-50 S-I rectorite RAr-1, 30-70 S-I ISCz-1, and nontronite NAu-2. Under an oxic condition, reduced clays (with a prefix r before mineral names) produced reactive oxygen species (ROS), and the antibacterial activity followed the order of rRAr-1 > rSWy-3 ≥ rNAu-2 ≫ rIMt-2 ≥ rISCz-1. The strongest antibacterial activity of rRAr-1 was contributed by a combination of •OH and Fe(IV) generated from structural Fe(II)/adsorbed Fe2+ and soluble Fe2+, respectively. Higher levels of lipid and protein oxidation, intracellular ROS accumulation, and membrane disruption were consistent with this antibacterial mechanism of rRAr-1. The antibacterial activity of other S-I clays depended on layer expandability, which determined the reactivity of structural Fe(II) and the production of •OH, with the expandable smectite being the most antibacterial and nonexpandable illite the least. Our results provide new insights into the antibacterial mechanisms of clay minerals.

Mechanistic Insight into Humic Acid-Enhanced Hydroxyl Radical Production from Fe(II)-Bearing Clay Mineral Oxygenation.

Hydroxyl radical (•OH) production by electron transfer from Fe(II)-bearing clay minerals to oxygen has been increasingly reported. However, the influence of ubiquitous coexisting humic acid (HA) on this process is poorly understood. Here, we investigated the effect of different HA on •OH production during the oxygenation of reduced nontronite NAu-2 (rNAu-2), montmorillonite, and sediment. Results showed that HA could enhance •OH production, and the enhancement was related to the content of reactive Fe(II) in rNAu-2 and the electron-accepting capacity of HA. Coexisting HA leads to a new electron-transfer pathway from Fe(II) in rNAu-2 to HA (instead of the HA-Fe complex) and then to O2, changing the first step of O2 reduction from one- to two-electron transfer process with H2O2 as the main intermediate. Reduced HA decomposes H2O2 to •OH at a higher yield (13.8%) than rNAu-2 (8.8%). Modeling results reveal that the HA-mediated electron-transfer pathway contributes to 12.6-70.2% of H2O2 generation and 13.2-62.1% of •OH formation from H2O2 decomposition, with larger contributions at higher HA concentrations (5-100 mg C/L). Our findings implicate that HA-mediated electron transfer can expand the area of •OH production from the mineral surface to the aqueous phase and increase the yield of •OH production.

Lignin-enhanced reduction of structural Fe(III) in nontronite: Dual roles of lignin as electron shuttle and donor

Lignin is a major component of plant-derived soil organic matter (SOM) in soils and sediments. Fe-bearing clay minerals are widely distributed in these environments and often co-exist with lignin. While previous studies have reported the electron shuttling and donating roles of certain redox-active SOM in the dissimilatory reduction of structural Fe(III) in Fe-bearing clay minerals, the role of lignin in this process remains unknown. Here we studied this role by incubating an Fe-rich smectite (nontronite NAu-2) with two types of lignin (soluble and insoluble) in the absence and presence of an Fe(III)-reducing bacterium Shewanella putrefaciens CN32 under anaerobic condition. Lactate was added in some experiments as an extra electron donor. The results demonstrated that both soluble and insoluble lignins abiotically reduced structural Fe(III) in NAu-2. The reduction extent was proportional to lignin concentration. After abiotic reaction, lignin served as either electron shuttle or electron donor in the presence of CN32: (1) When lactate was present, lignin served as an electron shuttle to enhance the rate of Fe(III) reduction; (2) When lactate was absent, lignin served as an electron donor for Fe(III) reduction. Although the ultimate biotic Fe(III) reduction extents were similar in the presence of either soluble or insoluble lignin, the reduction rates with soluble lignin were higher than those with insoluble lignin, likely owing to their different electron transfer mechanisms. After interaction with NAu-2 and/or CN32, soluble lignin structure largely remained intact, but with some decreases of humic/fulvic acid-like and protein-like compounds, aromatic functional groups (e.g., CH, CO, COOH), and aliphatic/aromatic compounds. An increase of semiquinone-like organic radicals was observed after lignin interaction with NAu-2. These chemical changes of lignin were likely coupled with the reduction of structural Fe(III) in nontronite. Upon reduction, the nontronite did not display much dissolution and mineral transformation. The findings of this study provide insights into the role of lignin in promoting mineral-microbe interactions and have significant implications for coupled Fe and C biogeochemical cycles in soils and sediments.

Molecular Determination of Organic Adsorption Sites on Smectite during Fe Redox Processes Using ToF-SIMS Analysis.

Turnover of soil organic carbon (SOC) is strongly affected by a balance between mineral protection and microbial degradation. However, the mechanisms controlling the heterogeneous and preferential adsorption of different types of SOC remain elusive. In this work, the heterogeneous adsorption of humic substances (HSs) and microbial carbon (MC) on a clay mineral (nontronite NAu-2) during microbial-mediated Fe redox cycling was determined using time-of-flight secondary ion mass spectrometry (ToF-SIMS). The results revealed that HSs pre-adsorbed on NAu-2 would partially inhibit structural modification of NAu-2 by microbial Fe(III) reduction, thus retarding the subsequent adsorption of MC. In contrast, NAu-2 without precoated HSs adsorbed a significant amount of MC from microbial polysaccharides as a result of Fe(III) reduction. This was attributed to the deposition of a thin Al-rich layer on the clay surface, which provided active sites for MC adsorption. This study provides direct and detailed molecular evidence for the first time to explain the preferential adsorption of MC over HSs on the surface of clay minerals in iron redox processes, which could be critical for the preservation of MC in soil. The results also indicate that ToF-SIMS is a unique tool for understanding complex organic-mineral-microbe interactions.

Anaerobic oxidation of arsenite by bioreduced nontronite

The redox state of arsenic controls its toxicity and mobility in the subsurface environment. Understanding the redox reactions of arsenic is particularly important for addressing its environmental behavior. Clay minerals are commonly found in soils and sediments, which are an important host for arsenic. However, limited information is known about the redox reactions between arsenic and structural Fe in clay minerals. In this study, the redox reactions between As(III)/As(V) and structural Fe in nontronite NAu-2 were investigated in anaerobic batch experiments. No oxidation of As(III) was observed by the native Fe(III)-NAu-2. Interestingly, anaerobic oxidation of As(III) to As(V) occurred after Fe(III)-NAu-2 was bioreduced. Furthermore, anaerobic oxidization of As(III) by bioreduced NAu-2 was significantly promoted by increasing Fe(III)-NAu-2 reduction extent and initial As(III) concentrations. Bioreduction of Fe(III)-NAu-2 generated reactive Fe(III)-O-Fe(II) moieties at clay mineral edge sites. Anaerobic oxidation of As(III) was attributed to the strong oxidation activity of the structural Fe(III) within the Fe(III)-O-Fe(II) moieties. Our results provide a potential explanation for the presence of As(V) in the anaerobic subsurface environment. Our findings also highlight that clay minerals can play an important role in controlling the redox state of arsenic in the natural environment.

Microbial mobilization of arsenic from iron-bearing clay mineral through iron, arsenate, and simultaneous iron-arsenate reduction pathways

Clay minerals are an important host for arsenic in many arsenic-affected areas. The role of bioreduction of structural Fe(III) in clay minerals in the mobilization of arsenic from clay minerals, however, still remains unclear. In this study, Fe(III) reducing bacterium, As(V) reducing bacterium, and Fe(III)-As(V) reducing bacterium were employed to investigate the possible bioreduction pathways for arsenic release from Nontronite NAu-2. Results demonstrated that microbial reduction controlled arsenic mobilization from NAu-2 through Fe(III), As(V), and simultaneous Fe(III)-As(V) reduction pathways. Although the bioreduction of structural Fe(III) led to a negligible dissolution of NAu-2, it triggered a significant release of arsenic from NAu-2. The bioreduction of tetrahedral Fe(III) initiated the release of As(V), and the further bioreduction of octahedral Fe(III) induced the release of As(III) in NAu-2. In addition, bioreduction of As(V) resulted in the desorption and transformation of As(V) from NAu-2. Simultaneous bioreduction of Fe(III) and As(V) led to an almost complete release of As(V) from NAu-2. These findings suggest that simultaneous Fe(III)-As(V) reduction was the dominant pathway governing As(V) release from NAu-2, while structural Fe(III) reduction controlled As(III) release from NAu-2. Therefore, the bioreduction of iron-bearing clay minerals has a great potential for arsenic mobilization in the subsurface environment.

Promotion of Microbial Oxidation of Structural Fe(II) in Nontronite by Oxalate and NTA.

Iron redox cycling occurs extensively in soils and sediments. Previous research has focused on microbially mediated redox cycling of aqueous Fe. At circumneutral pH, most Fe occurs in solid phase, where Fe and organic ligands interact closely. However, the role of organic ligands in microbial oxidation of solid-phase Fe(II) is not well understood. Here, we incubated reduced nontronite NAu-2 (rNAu-2) with an iron-oxidizing bacterium and in the presence of oxalate and nitrilotriacetic acid. These ligands significantly enhanced the rate and extent of microbial oxidation of structural Fe(II) in rNAu-2. Aqueous and solid-phase analyses, coupled with biogeochemical modeling, revealed a pathway for ligand-enhanced bio-oxidation of solid-phase Fe(II): (1) dissolution of rNAu-2 to form aqueous Fe(II)-ligand complex; (2) bio-oxidation to Fe(III)-ligand complex; (3) rapid reduction of Fe(III)-ligand complex to Fe(II)-ligand complex by structural Fe(II) in rNAu-2. In this process, the Fe(II)-ligand and Fe(III)-ligand complexes effectively serve as electron shuttle to expand the bioavailable pool of solid-phase Fe(II). These results have important implications for a better understanding of the bioavailability and reactivity of solid-phase Fe pool in the environment.

Mechanisms of Enhanced Antibacterial Activity by Reduced Chitosan-Intercalated Nontronite

Previous studies have documented the antibacterial activity of certain iron-containing clays. However, the repulsion between negatively charged bacteria and the clay surface makes this process inefficient. The objective of this study is to improve the bactericidal efficiency of clays by reversing their surface charge from negative to positive. To achieve this objective, positively charged chitosan, a nontoxic and biodegradable polymer, was intercalated into nontronite NAu-2. Chitosan-intercalated NAu-2 (C-NAu-2) was chemically reduced to obtain reduced C-NAu-2 (rC-NAu-2). Relative to reduced nontronite (rNAu-2), the antibacterial activity of rC-NAu-2 is higher and more persistent over a pH range of 6-8. The close spatial association between positively charged rC-NAu-2 and negatively charged bacteria increases the chances of cell membrane attack by extracellular ROS, the influx of soluble Fe2+ into the bacterial cell, and the yield of intracellular ROS. All these factors contribute to the enhanced antibacterial activity of rC-NAu-2. In contrast to rNAu-2 treated E. coli cells, where membrane damage and intracellular ROS/Fe accumulation are restricted to the polar regions, the close bacteria-clay association in rC-NAu-2 results in nonselective membrane damage and more uniform intracellular ROS/Fe distribution across whole bacterial cells. These results advance the antibacterial model by highlighting the importance of bacteria-clay interactions to the antibacterial activity of Fe-bearing clays.

Surface-bound radical control rapid organic contaminant degradation through peroxymonosulfate activation by reduced Fe-bearing smectite clays

Heterogeneously activated peroxymonosulfate (PMS)-based advanced oxidation technologies (AOTs) have received increasing attention in contaminated water remediation. However, PMS activation by reduced clay minerals (e.g., reduced Fe-bearing smectite clays) has rarely been explored. Herein, PMS decomposition by reduced Fe-bearing smectite clays was investigated, and the hydroxyl radical (OH) and sulfate radical (SO4−) formation mechanisms were elucidated. Reduced nontronite NAu-2 (R-NAu-2) activated PMS efficiently to induce rapid degradation of diethyl phthalate (DEP) within 30 s. Mössbauer spectroscopy, FTIR and XPS analyses substantiated that distorted trans-coordinated Fe(II)Fe(II)Fe(II)OH entities were mainly responsible for rapid electron transfer to regenerate clay surface Fe(II) for PMS activation. Chemical probe, radical quenching, and electron paramagnetic resonance (EPR) results confirmed that OH and SO4− were mainly bound to the clay surface rather than in bulk solution, which resulted in the rapid degradation of organic compounds such as DEP, sulfamethoxazole, phenol, chlortetracycline and benzoic acid. Anions such as Cl− and NO3− had a limited effect on DEP degradation, while HCO3− inhibited the DEP degradation due to the increase of reaction pH. This study provides a new PMS activation strategy using reduced Fe-bearing smectite clays that will contribute to rapid degradation of organic contaminants using PMS-based AOTs.

Effect of Coexisting Fe(III) (oxyhydr)oxides on Cr(VI) Reduction by Fe(II)-Bearing Clay Minerals.

Fe(II)-bearing clay minerals are important electron sources for Cr(VI) reduction in subsurface environments. However, it is not clear how iron (oxyhydr)oxides impact Cr(VI) reduction by Fe(II)-bearing clays as the two minerals can coexist in soil and sediment aggregates. This study investigated Cr(VI) reduction in the mixed suspensions of reduced nontronite NAu-2 (rNAu-2) and ferrihydrite (Fe(II)/Cr(VI) = 3:1). When the mineral premixing time increased from 0 to 72 h, Cr(VI) reduction was accelerated prominently in the initial stage, while Cr(VI) sorption was inhibited drastically. Mineral premixing led to electron transfer from structural Fe(II) in rNAu-2 to ferrihydrite with formation of reactive-surface-associated Fe(II), which catalyzed ferrihydrite transformation to lepidocrocite. Reactive-surface-associated Fe(II) accelerated Cr(VI) reduction initially, and ferrihydrite transformation to lepidocrocite was responsible for the inhibited sorption. When the reactive-surface-associated Fe(II) was consumed in the initial stage, the Cr(VI) reduction rate decreased dramatically due to the limitation of slow electron transfer from structural Fe(II) in rNAu-2 to surface-reactive sites. The main reduction sites shifted from rNAu-2 to ferrihydrite/lepidocrocite when rNAu-2 coexisted with ferrihydrite. Our findings demonstrate that electron transfer between minerals has important implications for Cr(VI) and other high-valence contaminant reduction by Fe(II)-bearing clay minerals in subsurface environments.

Anoxic storage regenerates reactive Fe(II) in reduced nontronite with short-term oxidation

Fe(II)-bearing clay minerals represent a large electron source under redox-dynamic systems. Although redox reactions between Fe(II)-bearing clay minerals and oxidants (such as O2) have been widely investigated in redox-dynamic systems, one important process that remains unexplored is the response of Fe(II) reactivity in clay minerals to long-term anoxic storage following short-term oxidation. Here we used reduced nontronite NAu-2 as a model Fe-bearing clay mineral, exposed its suspension to air for oxidation in a short time (1, 3 and 5 h), and then stored it under anoxic conditions without any exogenous oxidants for a long time (0–120 h). After different time of anoxic storage, we evaluated the reactivity of Fe(II) by air oxidation, measured the Fe(II) fractions by phosphate extraction and characterized the variation of Fe(II) by X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and Mössbauer. We found that anoxic storage increased prominently the rate of Fe(II) oxidation by O2 and the resultant production of hydroxyl radicals, and the increase was more obvious for the NAu-2 with a longer time of anoxic storage. Both phosphate extraction and XPS results revealed that the reactive Fe(II) associated with edge sites was regenerated and the regeneration increased with the increase in anoxic storage time. FTIR and Mössbauer spectra identified the rearrangements of NAu-2 structure during anoxic storage. The mechanism of reactive Fe(II) regeneration by anoxic storage was attributed to the increase in reactive Fe(II) associated with edge sites, which was most likely induced by the electron transfer from Fe(II) at the interior sites to Fe(III) at the edge sites in the octahedral sheets of nontronite. The driving force for electron transfer was presumed to be the redox potential gradient between edge and interior sites because of different Fe(II)/Fe(III) ratios. Our findings improve the understanding of Fe(II)/Fe(III) interconversion as well as the resultant reducing impact on substance transformation induced by Fe(II)-bearing clay minerals under redox-dynamic systems.

Iron(III) minerals and anthraquinone-2,6-disulfonate (AQDS) synergistically enhance bioreduction of hexavalent chromium by Shewanella oneidensis MR-1

Bioreduction of hexavalent chromium (Cr(VI)) to sparingly soluble trivalent chromium (Cr(III)) is a strategy for the remediation of Cr(VI) contaminated sites. However, its application is limited due to the slow bioreduction process. Here we explored the potential synergistic enhancement of iron(III) minerals (nontronite NAu-2, ferrihydrite, and goethite) and electron shuttle anthraquinone-2,6-disulfonate (AQDS) on the bioreduction of Cr(VI) by Shewanella oneidensis MR-1. AQDS alone increased the bioreduction rate of Cr(VI) by accelerating electron transfer from MR-1 to Cr(VI). Iron minerals alone did not increase the bioreduction rate of Cr(VI), where the electron transfer from MR-1 to Fe(III) minerals was inhibited due to the toxicity of Cr(VI) to MR-1. AQDS plus NAu-2 or ferrihydrite significantly enhanced the bioreduction rate of Cr(VI) as compared to AQDS or NAu-2/ferrihydrite alone, demonstrating that AQDS plus NAu-2/ferrihydrite had the synergistic effect on bioreduction of Cr(VI). Synergy factor (kcells+Fe+AQDS/(kcells+Fe + kcells+AQDS)) was used to quantify the synergistic effect of AQDS and iron minerals on the bioreduction of Cr(VI). The synergy factors of AQDS plus NAu-2 were 2.09–4.63 (three Cr(VI) spikes), and the synergy factors of AQDS plus ferrihydrite were 1.89–4.61 (two Cr(VI) spikes). In the presence of Cr(VI), AQDS served as the electron shuttle between MR-1 and iron minerals, facilitating the reduction of Fe(III) minerals to Fe(II). The synergistic enhancement of AQDS and NAu-2/ferrihydrite was attributed to the generated Fe(II), which could quickly reduce Cr(VI) to Cr(III). Our results provide an attractive strategy to strengthen the bio-immobilization of Cr(VI) at iron-rich contaminated sites through the synergistic enhancement of iron(III) minerals and electron shuttle.

Reduction of structural Fe(III) in nontronite by thermophilic microbial consortia enriched from hot springs in Tengchong, Yunnan Province, China

Iron redox cycling provides one of the most important energy sources supporting microorganisms in terrestrial hot springs. In order to understand microbial Fe(III) reduction processes in hot springs, Fe(III)-reducing bacterial consortia were successfully retrieved from seven acidic hot springs in Tengchong County, Yunnan Province, China (a temperature range from 40.6°C to 74.7°C and a pH range from 2.64 to 4.72). These Fe(III)-reducing consortia were mainly composed of sulfate reducing bacteria (SRB, including Desulfotomaculum, Thermodesulfobacterium and Desulfomicrobium) and organic matter fermenting bacteria (Thermoanaerobacterium and Thermoanaerobacter), with each consortia having specific composition depending on the physio-chemical conditions of the studied hot springs. The obtained consortia were capable of dissimilatory reduction of Fe(III) in nontronite NAu-2 (ferruginous smectite) with 20.8% to 29.8% reduction extent and initial reduction rate of 0.011–0.023mMFe(III)/gNAu-2/h. These extents and rates were comparable to those by mesophilic Fe-reducers in the presence of electron shuttles. Both high temperature and the possible presence of electron-shuttles in hot springs may have accounted for these observed high reduction extents and rates. Various secondary minerals formed during NAu-2 bioreduction, including Fe sulfides and oxides, silicates, and silica. In one spring, illitization and kaolinization of smectite were observed, suggesting that Fe-reducers play important roles in mineral transformation in geothermal environments.

Reduced Iron-Containing Clay Minerals as Antibacterial Agents.

Previous work documented the general antibacterial mechanism of iron containing clays that involved hydroxyl radical (•OH) production from soluble Fe2+, and attack of cell membrane and intracellular proteins. Here we explore the role of clay structural Fe(II) in •OH production at near neutral pH and identify a lipid involved in the antibacterial process. Structural Fe(III) in nontronite NAu-2 was reduced (rNAu-2) and E. coli, a model bacterium, was exposed to rNAu-2 in oxic suspension. The antibacterial activity of rNAu-2 was dependent on pH and Fe(II) concentration, where E. coli were completely killed at pH 6, but survived at pH 7 and 8. In the presence of a •OH scavenger or in anaerobic atmosphere, E. coli survived better, suggesting that cell death may be caused by •OH generated from oxidation of structural Fe(II) in rNAu-2. In-situ imaging revealed damage of a membrane lipid, cardiolipin, in the polar region of E. coli cells, where reactive oxygen species and redox-active labile Fe were enriched. Our results advance the previous antibacterial model by demonstrating that the structural Fe(II) is the primary source of •OH, which damages cardiolipin, triggers the influx of soluble Fe2+ into the cell, and ultimately leads to cell death.

Microbial reduction of Fe(III)-bearing clay minerals in the presence of humic acids

Both Fe(III)-bearing clay minerals and humic acids (HAs) are abundant in the soils and sediments. Previous studies have shown that bioreduction of structural Fe(III) in clay minerals could be accelerated by adding anthraquinone compound as a redox-active surrogate of HAs. However, a quinoid analogue could not reflect the adsorption and complexation properties of HA, and little is known about the effects of real HAs at environmental concentration on bioreduction of clay minerals. Here, it was shown that 10–200 mg l−1 of natural or artificially synthesized HAs could effectively stimulate the bioreduction rate and extent of Fe(III) in both iron-rich nontronite NAu-2 and iron-deficient montmorillonite SWy-2. After adsorption to NAu-2, electron-transfer activities of different HA fractions were compared. Additionally, Fe(II) complexation by HAs also contributed to improvement of clay-Fe(III) bioreduction. Spectrosopic and morphological analyses suggested that HA addition accelerated the transformation of NAu-2 to illite, silica and siderite after reductive dissolution.