Low and high acetate amendments are equally as effective at promoting complete dechlorination of trichloroethylene (TCE)

Experiments with trichloroethylene-contaminated aquifer material demonstrated that TCE, cis-DCE, and VC were completely degraded with concurrent Fe(III) or Fe(III) and sulfate reduction when acetate was amended at stoichiometric concentration; competing TEAPs did not inhibit ethene production. Adding 10× more acetate did not increase the rate or extent of TCE reduction, but only increased methane production. Enrichment cultures demonstrated that ~90 μM TCE or ~22 μM VC was degraded primarily to ethene within 20 days with concurrent Fe(III) or Fe(III) + sulfate reduction. The dechlorination rates were comparable between the low and high acetate concentrations (0.36 vs 0.34 day^?1, respectively), with a slightly slower rate in the 10× acetate amended incubations. Methane accumulated to 13.5 (±0.5) μmol/tube in the TCE-degrading incubations with 10× acetate, and only 1.4 (±0.1) μmol/tube with low acetate concentration. Methane accumulated to 16 (±1.5) μmol/tube in VC-degrading enrichment with 10× acetate and 2 (±0.1) μmol/tube with stoichiometric acetate. The estimated fraction of electrons distributed to methanogenesis increased substantially when excessive acetate was added. Quantitative PCR analysis indicated that 10× acetate did not enhance Dehalococcoides biomass but rather increased the methanogen abundance by nearly one order of magnitude compared to that with stoichiometric acetate. The data suggest that adding low levels of substrate may be equally if not more effective as high concentrations, without producing excessive methane. This has implications for field remediation efforts, in that adding excess electron donor may not benefit the reactions of interest, which in turn will increase treatment costs without direct benefit to the stakeholders.     

INFLUENCE OF SOIL PROPERTIES AND PHOSPHATE ADDITION ON ARSENIC UPTAKE FROM POLLUTED SOILS BY VELVETGRASS (HOLCUS LANATUS)

Four kinds of soil material were used in a pot experiment with velvetgrass (Holcus lanatus). Two unpolluted soils: sand (S) and loam (L) were spiked with sodium arsenite (As III) and arsenate (As V), to obtain total arsenic (As) concentrations of 500 mg As kg^?1. Two other soils (ZS I, ZS III), containing 3320 and 5350 mg As kg^?1, were collected from Zloty Stok where gold and arsenic ores were mined and processed for several centuries. The effects of phosphate addition on plants growth and As uptake were investigated. Phosphate was applied to soils in the form of NH₄H₂PO₄ at the rate 0.2 g P/kg. Average concentrations of arsenic in the shoots of velvetgrass grown in spiked soils S and L without P amendment were in the range 18–210 mg As kg^?1 d.wt., whereas those in plants grown on ZS I and ZS II soils were considerably lower, and varied in the range 11–52 mg As kg^?1 d.wt. The addition of phosphate caused a significant increase in plant biomass and therefore the total amounts of As taken up by plants, however, the differences in As concentrations in the shoots of velvetgrass amended and non-amended with phosphate were not statistically significant.     

Interannual Variability of Low-Molecular Metabolite Composition in <Emphasis Type="Italic">Ceratophyllum demersum</Emphasis> (Ceratophyllaceae) from a Floodplain Lake with a Changeable Trophic Status

The regularities that shape the composition of low molecular weight organic compounds (LMWOCs) in aquatic macrophytes in response to aquatic environment alterations remain poorly characterized. The aim of the present study consists of a comparative interannual investigation into LMWOC composition in rigid hornwort (Ceratophyllum demersum L.) from a Volga-Akhtuba floodplain lake with a variable trophic state. A high variability of LMWOC composition and individual compound levels in hornwort is detected as different trophic states of the water body are analyzed. Active allelochemicals are the predominant LMWOCs in the case of a “macrophytic” mesotrophic state of the lake, with fatty acids (the free fatty acid fraction) apparently being the most important in this group. Hornwort LMWOC composition in the case of a “cyanobacterial” eutrophic type of lake development is characterized by the predomination of compounds that enhance the protective reactions (manool being the most important) under the conditions of suppression by cyanobacteria, which is also manifested as an almost twofold decrease in the overall intensity of organiccompound biosynthesis.     

Abundance and Diversity of Sulfate-Reducing Bacteria in High Arsenic Shallow Aquifers

The abundance, diversity, and relative distribution of sulfate-reducing bacteria (SRB) in high arsenic (As) groundwater aquifers of Hangjinhouqi County in the Hetao Basin, Inner Mongolia was investigated using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction (qPCR) analysis of dsrB genes (encoding dissimilatory sulfite reductase beta-subunit). DGGE results revealed that SRB populations were diverse, but were mainly composed of Desulfotomaculum, Desulfobulbus, Desulfosarcina, and Desulfobacca. The abundance of Desulfobulbus was positively correlated with the ratio of Fe(II)/Fe(III). Although qPCR results showed that the dsrB gene abundance in groundwater samples ranged from below detection to 4.9 × 10⁶ copies/L, and the highest percentage of dsrB gene copies to bacterial 16S rRNA gene copies was 2.1%. Geochemical analyses showed that As(III) content and the ratio of Fe(II) to Fe(III) increased with total As, while sulfate concentrations decreased. Interestingly, the dsrB gene abundance was positively correlated with As concentrations. These results indicate that sulfate reduction occurs simultaneously with As and Fe reduction, and might result in increased As release and mobilization when As is not incorporated into iron sulfides. This study improves our understanding of SRB and As cycling in high As groundwater systems.     

SORPTION OF FERROUS AND FERRIC IRON BY EXTRACELLULAR POLYMERIC SUBSTANCES (EPS) FROM ACIDOPHILIC BACTERIA

The sorption of Fe(II) and Fe(III) by extracellular polymeric substances (EPS) of acidophilic bacteria Acidiphilium 3.2Sup(5) and Acidithiobacillus ferrooxidans, harvested from the ecosystem of the Tinto River (Huelva, Spain), was investigated. EPS from mixed cultures of both bacteria (EPS_mixed) and pure cultures of A. 3.2Sup(5) (EPS_pure) were extracted with ethylenediamine tetraacetic acid (EDTA) and were characterized by Fourier-transform infrared (FTIR), electron photoemission (XPS), x-ray diffraction (DRX), and energy dispersive x-ray (EDX) spectroscopy and scanning electron microscopy (SEM). EPS _pure were loaded, in sorption tests, with Fe(II) and Fe(III). The results obtained indicate that the biochemical composition and structure of EPS_mixed was very similar to that of EPS_pure. Besides, results indicate that EPS_mixed adsorbed Fe(II) and Fe(III) by preferential interaction with the carboxyl group, which favored the formation of Fe(II)/Fe(III) oxalates. These species were also formed in EPS_pure loaded with Fe(II)/Fe(III). All this behavior suggested that the sorption of iron by EPS_mixed was similar to sorption of EPS_pure, which fitted the Freundlich model. Thus, the iron uptake of EPS_mixed reached 516.7 ± 23.4 mg Fe/g-EPS at an initial concentration of 2.0 g/L of Fe_total and Fe(II)/Fe(III) ratio of 1.0.     

Bacterial Community Structure and Functional arrA Gene Diversity Associated with Arsenic Reduction and Release in an Industrially Contaminated Soil

This study aimed at evaluating potential arsenic (As) mobility in an industrially contaminated soil (64 mg/kg of As) of the Meuse River basin, and at identifying key bacterial groups that drive soil As dynamics. Both speciation and release of As from this soil was followed under anaerobic conditions using a laboratory batch experiment. In the presence of exogenous carbon sources, As^V initially present in the soil matrix and/or adsorbed on synthetic hydrous ferric oxides were solubilized and mainly reduced to As^III by indigenous soil microflora. After a 1-month incubation period in these biotic conditions, As^III accounted for 80–85% of the total dissolved As and more than 60% of the solid-phase As. Bacterial community structure (i.e., 16S rDNA-based capillary electrophoresis single-strand conformation polymorphism profiles) changed with incubation time and As amendment. The detection of distantly related arsenate respiratory reductase genes (arrA), as functional markers of As^V respirers, indicates that novel dissimilatory As^V-reducing bacteria may be involved in As biotransformation and mobility in anoxic soils. Since As and iron were concomitantly released, a crucial role of indirect As-mobilizing bacteria on As behavior was also revealed. Our results show that the majority of As within the soil matrix was bioavailable and bioaccessible for heterotrophic As^V reduction to As^III, which may increase As toxicity and mobility in the contaminated soils.     

Oxygen Dependency of Neutrophilic Fe(II) Oxidation by Leptothrix Differs from Abiotic Reaction

Neutrophilic Fe(II) oxidizing microorganisms are found in many natural environments. It has been hypothesized that, at low oxygen concentrations, microbial iron oxidation is favored over abiotic oxidation. Here, we compare the kinetics of abiotic Fe(II) oxidation to oxidation in the presence of the bacterium Leptothrix cholodnii Appels isolated from a wetland sediment. Rates of Fe(II) oxidation were determined in batch experiments at 20°C, pH 7 and oxygen concentrations between 3 and 120 μmol/l. The reaction progress in experiments with and without cells exhibited two distinct phases. During the initial phase, the oxygen dependency of microbial Fe(II) oxidation followed a Michaelis-Menten rate expression (K_M = 24.5 ± 10 μmol O₂/l, v_max = 1.8 ± 0.2 μmol Fe(II)/(l min) for 10⁸ cells/ml). In contrast, abiotic rates increased linearly with increasing oxygen concentrations. At similar oxygen concentrations, initial Fe(II) oxidation rates were faster in the experiments with bacteria. During the second phase, the accumulated iron oxides catalyzed further oxidative iron precipitation in both abiotic and microbial reaction systems. That is, abiotic oxidation also dominated the reaction progress in the presence of bacteria. In fact, in some experiments with bacteria, iron oxidation during the second phase proceeded slower than in the absence of bacteria, possibly due to an inhibitory effect of extracellular polymeric substances on the growth of Fe(III) oxides. Thus, our results suggest that the competitive advantage of microbial iron oxidation in low oxygen environments may be limited by the autocatalytic nature of abiotic Fe(III) oxide precipitation, unless the accumulation of Fe(III) oxides is prevented, for example, through a close coupling of Fe(II) oxidation and Fe(III) reduction.     

Ecophysiology and the energetic benefit of mixotrophic Fe(II) oxidation by various strains of nitrate-reducing bacteria

In order to assess the importance of nitrate-dependent Fe(II) oxidation and its impact on the growth physiology of dominant Fe oxidizers, we counted these bacteria in freshwater lake sediments and studied their growth physiology. Most probable number counts of nitrate-reducing Fe(II)-oxidizing bacteria in the sediment of Lake Constance, a freshwater lake in Southern Germany, yielded about 10⁵ cells mL⁻¹ of the total heterotrophic nitrate-reducing bacteria, with about 1% (10³ cells mL⁻¹) of nitrate-reducing Fe(II) oxidizers. We investigated the growth physiology of Acidovorax sp. strain BoFeN1, a dominant nitrate-reducing mixotrophic Fe(II) oxidizer isolated from this sediment. Strain BoFeN1 uses several organic compounds (but no sugars) as substrates for nitrate reduction. It also reduces nitrite, dinitrogen monoxide, and O₂, but cannot reduce Fe(III). Growth experiments with cultures amended either with acetate plus Fe(II) or with acetate alone demonstrated that the simultaneous oxidation of Fe(II) and acetate enhanced growth yields with acetate alone (12.5 g dry mass mol⁻¹ acetate) by about 1.4 g dry mass mol⁻¹ Fe(II). Also, pure cultures of Pseudomonas stutzeri and Paracoccus denitrificans strains can oxidize Fe(II) with nitrate, whereas Pseudomonas fluorescens and Thiobacillus denitrificans strains did not. Our study demonstrates that nitrate-dependent Fe(II) oxidation contributes to the energy metabolism of these bacteria, and that nitrate-dependent Fe(II) oxidation can essentially contribute to anaerobic iron cycling.     

The design of long-term effective uranium bioremediation strategy using a community metabolic model

Acetate amendment at uranium contaminated sites in Rifle, CO. leads to an initial bloom of Geobacter accompanied by the removal of U(VI) from the groundwater, followed by an increase of sulfate‐reducing bacteria (SRBs) which are poor reducers of U(VI). One of the challenges associated with bioremediation is the decay in Geobacter abundance, which has been attributed to the depletion of bio‐accessible Fe(III), motivating the investigation of simultaneous amendments of acetate and Fe(III) as an alternative bioremediation strategy. In order to understand the community metabolism of Geobacter and SRBs during artificial substrate amendment, we have created a genome‐scale dynamic community model of Geobacter and SRBs using the previously described Dynamic Multi‐species Metabolic Modeling framework. Optimization techniques are used to determine the optimal acetate and Fe(III) addition profile. Field‐scale simulation of acetate addition accurately predicted the in situ data. The simulations suggest that batch amendment of Fe(III) along with continuous acetate addition is insufficient to promote long‐term bioremediation, while continuous amendment of Fe(III) along with continuous acetate addition is sufficient to promote long‐term bioremediation. By computationally minimizing the acetate and Fe(III) addition rates as well as the difference between the predicted and target uranium concentration, we showed that it is possible to maintain the uranium concentration below the environmental safety standard while minimizing the cost of chemical additions. These simulations show that simultaneous addition of acetate and Fe(III) has the potential to be an effective uranium bioremediation strategy. They also show that computational modeling of microbial community is an important tool to design effective strategies for practical applications in environmental biotechnology. Biotechnol. Bioeng. 2012; 109: 2475–2483. © 2012 Wiley Periodicals, Inc.     

Application of Heavy Metal Rich Tannery Sludge on Sustainable Growth,Yield and Metal Accumulation by Clarysage (Salvia sclarea L.)

A field experiment was conducted to evaluate the effective utilization of tannery sludge for cultivation of clarysage (Salvia sclarea) at CIMAP research farm, Lucknow, India during the year 2012–2013. Six doses (0, 20, 40, 60, 80, 100 tha^?1) of processed tannery sludge were tested in randomised block design with four replications. Results revealed that maximum shoot, root, dry matter and oil yield were obtained with application of 80 tha^?1of tannery sludge and these were 94, 113 and 61% higher respectively, over control. Accumulation of heavy metals (Cr, Ni, Fe, Pb) were relatively high in shoot portion of the plant than root. Among heavy metals, magnitude of chromium accumulation was higher than nickel, iron and lead in shoot as well as in root. Linalool, linalyl acetate and sclareol content in oil increased by 13,8 and 27% respectively over control, with tannery sludge application at 80 tha^?1. Heavy metals such as chromium, cadmium and lead content reduced in postharvest soil when compared to initial status. Results indicated that clarysage (Salvia sclarea) can be grown in soil amended with 80 tha^?1sludge and this can be a suitable accumulator of heavy metals for phytoremediation of metal polluted soils.     

Altitudinal Distribution of Ammonia-Oxidizing Archaea and Bacteria in Alpine Grassland Soils Along the South-Facing Slope of Nyqentangula Mountains,Central Tibetan Plateau

Nitrogen is a major limiting nutrient for the net primary production of terrestrial ecosystems, especially on sentinel alpine ecosystem. Ammonia oxidation is the first and rate-limiting step on nitrification process and is thus crucial to nitrogen cycle. To decipher climatic influence on ammonia oxidizers, their communities were characterized by qPCR and clone sequencing by targeting amoA genes (encoding the alpha subunit of ammonia mono-oxygenase) in soils from 7 sites over an 800 m elevation transect (4400–5200 m a.s.l.), based on “space-to-time substitution” strategy, on a steppe-meadow ecosystem located on the central Tibetan Plateau (TP). Archaeal amoA abundance outnumbered bacterial amoA abundance at lower altitude (<4800 m a.s.l.), but bacterial amoA abundance was greater in surface soils at higher altitude (≥4800 m a.s.l.). Archaeal amoA abundance decreased with altitude in surface soil, while its abundance stayed relatively stable and was mostly greater than bacterial amoA abundance in subsurface soils. Conversely, bacterial amoA abundance gradually increased with altitude at all three soil depths. Statistical analysis indicated that altitude-dependent factors, in particular pH and precipitation, had a profound effect on the abundance and community of ammonia-oxidizing bacteria, but only on the community composition of ammonia-oxidizing archaea along the altitudinal gradient. These findings imply that the shifts in the relative abundance and/or community structure of ammonia-oxidizing bacteria and archaea may result from the precipitation variation along the altitudinal gradient. Thus, we speculate that altitude-related factors (mainly precipitation variation combing changed pH), would play a vital role in affecting nitrification process on this alpine grassland ecosystem located at semi-arid area on TP.     

Alkaline iron(III) reduction by a novel alkaliphilic,halotolerant, <Emphasis Type="Italic">Bacillus</Emphasis> sp. isolated from salt flat sediments of Soap Lake

A halotolerant, alkaliphilic dissimilatory Fe(III)-reducing bacterium, strain SFB, was isolated from salt flat sediments collected from Soap Lake, WA. 16S ribosomal ribonucleic acid gene sequence analysis identified strain SFB as a novel Bacillus sp. most similar to Bacillus agaradhaerens (96.7% similarity). Strain SFB, a fermentative, facultative anaerobe, fermented various hexoses including glucose and fructose. The fructose fermentation products were lactate, acetate, and formate. Under fructose-fermenting conditions in a medium amended with Fe(III), Fe(II) accumulated concomitant with a stoichiometric decrease in lactate and an increase in acetate and CO₂. Strain SFB was also capable of respiratory Fe(III) reduction with some unidentified component(s) of Luria broth as an electron donor. In addition to Fe(III), strain SFB could also utilize nitrate, fumarate, or O₂ as alternative electron acceptors. Optimum growth was observed at 30°C and pH 9. Although the optimal salinity for growth was 0%, strain SFB could grow in a medium with up to 15% NaCl by mass. These studies describe a novel alkaliphilic, halotolerant organism capable of dissimilatory Fe(III) reduction under extreme conditions and demonstrate that Bacillus species can contribute to the microbial reduction of Fe(III) in environments at elevated pH and salinity, such as soda lakes.     

ARSENIC CHEMISTRY AND REMEDIATION IN HAWAIIAN SOILS

Past use of arsenical pesticides has resulted in elevated levels of arsenic (As) in some Hawaiian soils. Total As concentrations of 20–100 mg/kg are not uncommon, and can exceed 900 mg/kg in some lands formerly planted with sugarcane. With high contents of amorphous aluminosilicates and iron oxides in many Hawaii's volcanic ash-derived Andisols, a high proportion (25–30%) of soil As was associated with either these mineral phases or with organic matter. Less than 1% of the total As was water soluble or exchangeable. Furthermore, the soils can sorb As strongly: the addition of 1000 mg/kg as As (+5) resulted in only between 0.03 and 0.30 mg/L As in soil solution. In contrast, soils having more crystalline minerals (e.g., Oxisols) sorb less As and thus often contain less As. Phosphate fertilization increases As bioaccessibility, whereas the addition of Fe(OH)₃ decreases it. Brake fern (Pteris vittata L.) can be used to remove some soil As. Concentrations of As in fronds varied on average from 60 mg/kg when grown on a low-As Oxisol to 350 mg/kg when grown on a high-As Andisol. Ratios of leaf As to CaCl₂-extractable soil As were 12 and 222 for the Oxisol and Andisol, respectively.     

Acclimation of arsenic-resistant Fe(II)-oxidizing bacteria in aqueous environment

This study investigated the potential of the Fe(II)-oxidizing bacteria in removing arsenic in aqueous environment. The bacteria were isolated from the batch of tap water and rusty iron wires, and were acclimated to culture media amended with arsenic concentrations, gradually increasing from 100 μg L⁻¹ to 100 mg L⁻¹. Acclimated bacteria with enhanced arsenic tolerance were used to remove arsenic from the aqueous solution. These bacteria belonged to Pseudomonas species according to 16S rRNA gene sequences. Extracellular enzymes produced by these bacteria played important roles in microbial Fe(II) oxidization and Fe oxide precipitation. Moreover, these bacteria survived and propagated in high arsenic condition (100 mg L⁻¹ As). However, after As(III/V) acclimation, morphological characteristics of the bacteria showed some changes, e.g., shrinking of long bacillus. XRD (X-ray diffraction) patterns indicated that Fe oxide precipitations by Fe(II)-oxidizing bacteria in Fe-rich culture medium were poorly-crystallized ferrihydrites. Adsorption on the biogenic ferrihydrites greatly contributed to high arsenic removal efficiency of Fe(II)-oxidizing bacteria.     

Dependence of In Situ Bacterial Fe(II)-Oxidation and Fe(III)-Precipitation on Sequential Reactive Transport

A study was conducted to determine in situ rates of Fe(II) oxidation and Fe(III) precipitation along a 5.0 m reach of a ferruginous groundwater discharge zone under two distinct conditions; (i) the natural state featuring abundant flocculent mats of bacteriogenic iron oxides (BIOS) produced by Fe(II)-oxidizing bacteria, and (ii) after a manual washout of the streambed to remove the microbial mat. Examination of mat samples by differential interference contrast light microscopy revealed tangled meshworks of filamentous Leptothrix sheaths and helical Gallionella stalks intermixed with fine-grained hydrous ferric oxide (HFO) precipitates. The greatest accumulation of BIOS mat was 1.0 m downstream of the groundwater spring. Redox potential (Eh) increased sharply from 200 mV to over 300 mV over the last 2.0 m of the reach. Similarly, dissolved oxygen increased from < 10% saturation to almost 100% saturation over the last 2.0 m of the reach, whereas pH increased from 6.4 to 7.3. Pseudo-first-order rate constants determined on the basis of analytical solutions to sequential partial differential advection-dispersion-reaction equations for the linear Fe(II)→Fe(III)→HFO reaction network yielded in situ Fe(II) oxidation rate constants (k_ox) of 1.70 ± 0.20 min^?1 in natural conditions and 0.48 ± 0.14 min^?1 after washout. Corresponding Fe(III)-precipitation rates (k_p) before and after washout were 3.45 ± 0.10 min^?1 and 0.90 ± 0.01 min^?1, respectively. These values for k_ox and k_p are higher than estimates obtained from closed batch microcosm and laboratory experiments, underscoring the crucial dependence of in situ Fe(II) oxidation and Fe(III) precipitation rates on advective and dispersive mass transport. The results also highlight the influence that BIOS microbial mats exert on the reaction kinetics of the multiple heterogeneous reactions contributing not only to Fe(II)/Fe(III) redox transformations in groundwater discharge zones, but also the precipitation of HFO.     

Suppression of methanogenesis by dissimilatory Fe(III)-reducing bacteria in tropical rain forest soils: implications for ecosystem methane flux

Tropical forests are an important source of atmospheric methane (CH₄), and recent work suggests that CH₄ fluxes from humid tropical environments are driven by variations in CH₄ production, rather than by bacterial CH₄ oxidation. Competition for acetate between methanogenic archaea and Fe(III)‐reducing bacteria is one of the principal controls on CH₄ flux in many Fe‐rich anoxic environments. Upland humid tropical forests are also abundant in Fe and are characterized by high organic matter inputs, steep soil oxygen (O₂) gradients, and fluctuating redox conditions, yielding concomitant methanogenesis and bacterial Fe(III) reduction. However, whether Fe(III)‐reducing bacteria coexist with methanogens or competitively suppress methanogenic acetate use in wet tropical soils is uncertain. To address this question, we conducted a process‐based laboratory experiment to determine if competition for acetate between methanogens and Fe(III)‐reducing bacteria influenced CH₄ production and C isotope composition in humid tropical forest soils. We collected soils from a poor to moderately drained upland rain forest and incubated them with combinations of ¹³C‐bicarbonate, ¹³C‐methyl labeled acetate (¹³CH₃COO^?), poorly crystalline Fe(III), or fluoroacetate. CH₄ production showed a greater proportional increase than Fe²⁺ production after competition for acetate was alleviated, suggesting that Fe(III)‐reducing bacteria were suppressing methanogenesis. Methanogenesis increased by approximately 67 times while Fe²⁺ production only doubled after the addition of ¹³CH₃COO^?. Large increases in both CH₄ and Fe²⁺ production also indicate that the two process were acetate limited, suggesting that acetate may be a key substrate for anoxic carbon (C) metabolism in humid tropical forest soils. C isotope analysis suggests that competition for acetate was not the only factor driving CH₄ production, as ¹³C partitioning did not vary significantly between ¹³CH₃COO^? and ¹³CH₃COO^?+Fe(III) treatments. This suggests that dissimilatory Fe(III)‐reduction suppressed both hydrogenotrophic and aceticlastic methanogenesis. These findings have implications for understanding the CH₄ biogeochemistry of highly weathered wet tropical soils, where CH₄ efflux is driven largely by CH₄ production.     

Electron shuttle-stimulated RDX mineralization and biological production of 4-nitro-2,4-diazabutanal (NDAB) in RDX-contaminated aquifer material

The potential for extracellular electron shuttles to stimulate RDX biodegradation was investigated with RDX-contaminated aquifer material. Electron shuttling compounds including anthraquinone-2,6-disulfonate (AQDS) and soluble humic substances stimulated RDX mineralization in aquifer sediment. RDX mass-loss was similar in electron shuttle amended and donor-alone treatments; however, the concentrations of nitroso metabolites, in particular TNX, and ring cleavage products (e.g., HCHO, MEDINA, NDAB, and NH₄ ⁺) were different in shuttle-amended incubations. Nitroso metabolites accumulated in the absence of electron shuttles (i.e., acetate alone). Most notably, 40–50% of [¹⁴C]-RDX was mineralized to ¹⁴CO₂ in shuttle-amended incubations. Mineralization in acetate amended or unamended incubations was less than 12% within the same time frame. The primary differences in the presence of electron shuttles were the increased production of NDAB and formaldehyde. NDAB did not further degrade, but formaldehyde was not present at final time points, suggesting that it was the mineralization precursor for Fe(III)-reducing microorganisms. RDX was reduced concurrently with Fe(III) reduction rather than nitrate or sulfate reduction. Amplified 16S rDNA restriction analysis (ARDRA) indicated that unique Fe(III)-reducing microbial communities (β- and γ-proteobacteria) predominated in shuttle-amended incubations. These results demonstrate that indigenous Fe(III)-reducing microorganisms in RDX-contaminated environments utilize extracellular electron shuttles to enhance RDX mineralization. Electron shuttle-mediated RDX mineralization may become an effective in situ option for contaminated environments.     

Toxicity and bioremediation of As(III) and As(V) in the green microalgae Botryococcus braunii: A laboratory study

Worldwide threats of fuel shortages in the near future and climate change because of greenhouse gas emissions are posing severe challenges and therefore it is vital to search for sustainable ways of preventing the consequences. The dual use of microalgae for phycoremediation and biomass production for sustainable biofuel production is a viable choice. Phycoremediation of As(III) and As(V) ions using microalgae was investigated in a two-staged batch reactor. Accumulation and toxicity of inorganic arsenic forms (As(III) and As(V)) to green microalgae Botryococcus braunii depend on environmental factors. Dissolved oxygen and pH cycles did not significantly differ due to the absence or presence of arsenic (either As(III) or As(V)) ions in the culture. Monod model was utilized for representing the growth kinetics of microalgae in pure media containing various concentrations of nitrate ions. Maximum specific growth rate and saturation constant were found to be 0.14788 d^?1 and 0.00105 g/L, respectively. With the increase in concentration of phosphate in growth medium, the growth of microalgae increased. Media with NaCl (1.0 g/L) and NaHCO₃ (1 g/L) resulted in higher maximum biomass concentration. Effect of coexisting ions on phycoremediation of As(III) and As(V) ions using microalgae was studied.     

Taxonomical and functional microbial community dynamics in an Anammox-ASBR system under different Fe (III) supplementation

In the present study, we explored the metabolic versatility of anaerobic ammonium oxidation (anammox) bacteria in a variety of Fe (III) concentrations. Specifically, we investigated the impacts of Fe (III) on anammox growth rates, on nitrogen removal performance, and on microbial community dynamics. The results from our short-term experiments revealed that Fe (III) concentrations (0.04–0.10 mM) significantly promote the specific anammox growth rate from 0.1343 to 0.1709 d^?1. In the long-term experiments, the Anammox-anaerobic sequencing batch reactor (ASBR) was operated over 120 days and achieved maximum NH₄ ⁺-N, NO₂ ^?-N, and TN efficiencies of 90.98 ± 0.35, 93.78 ± 0.29, and 83.66 ± 0.46 %, respectively. Pearson’s correlation coefficients between anammox-(narG + napA), anammox-nrfA, and anammox-FeRB all exceeded r = 0.820 (p < 0.05), confirming an interaction and ecological association among the nitrogen and iron-cycling-related microbial communities. Illumina MiSeq sequencing indicated that Chloroflexi (34.39–39.31 %) was the most abundant phylum in an Anammox-ASBR system, followed by Planctomycetes (30.73–35.31 %), Proteobacteria (15.40–18.61 %), and Chlorobi (4.78–6.58 %). Furthermore, we found that higher Fe (III) supplementation (>0.06 mM) could result in the community succession of anammox species, in which Candidatus Brocadia and Candidatus Kuenenia were the dominant anammox bacteria species. Combined analyses indicated that the coupling of anammox, dissimilatory nitrogen reduction to ammonium, and iron reduction accounted for nitrogen loss in the Anammox-ASBR system. Overall, the knowledge gained in this study provides novel insights into the microbial community dynamics and metabolic potential of anammox bacteria under Fe (III) supplementation.     

Viral and bacterial community responses to stimulated Fe(III)-bioreduction during simulated subsurface bioremediation

The delivery of fermentable substrate(s) to subsurface environments stimulates Fe(III)-bioreduction and achieves detoxification of organic/inorganic contaminants. Although, much research has been conducted on the microbiology of such engineered systems at lab and field scales, little attention has been given to the phage-host interactions and virus community dynamics in these environments. The objective was to determine the responses of soil bacterial communities and viral assemblages to stimulated anaerobic Fe(III)-bioreduction following electron donor (e.g. acetate) addition. Microbial communities, including viral assemblages, were investigated after 60 days of Fe(III)-bioreduction in laboratory-scale columns continuously fed with acetate-amended artificial groundwater. Viral abundances were greatest in the influent section and decreased along the flow path. Acetate availability was important in influencing bacterial diversity, microbial interactions and viral abundance and community composition. The impact of acetate addition was most evident in the influent section of the columns. The increased relative abundance of Fe(III)-reducing bacteria coincided with an increase in viral abundance in areas of the columns exhibiting the most Fe(III) reduction. The genetic composition of viruses in these column sections also differed from the control column and distal sections of acetate-treated columns suggesting viral communities responded to biostimulated Fe(III)-bioreduction.