We studied the microbial functional and structural interactions between nitrate (NO(3)(-)) and perchlorate (ClO(4)(-)) reductions in the hydrogen (H(2))-based membrane biofilm reactor (MBfR). When H(2) was not limiting, ClO(4)(-) and NO(3)(-) reductions were complete, and the MBfR's biofilm was composed mainly of bacteria from the ε- and β-proteobacteria classes, with autotrophic genera Sulfuricurvum, Hydrogenophaga, and Dechloromonas dominating the biofilm. Based on functional-gene and pyrosequencing assays, Dechloromonas played the most important role in ClO(4)(-) reduction, while Sulfuricurvum and Hydrogenophaga were responsible for NO(3)(-) reduction. When H(2) delivery was insufficient to completely reduce both electron acceptors, NO(3)(-) reduction out-competed ClO(4)(-) reduction for electrons from H(2), and mixotrophs become important in the MBfR biofilm. β-Proteobacteria became the dominant class, and Azonexus replaced Sulfuricurvum as a main genus. The changes suggest that facultative, NO(3)(-)-reducing bacteria had advantages over strict autotrophs when H(2) was limiting, because organic microbial products became important electron donors when H(2) was severely limiting.

[1] This work presents a pore-scale biofilm model that solves the flow field using the lattice Boltzmann method, the concentration field of chemical species using the finite difference method, and biofilm development using the cellular automaton method. We adapt the model from a previous work and expand it by implementing biofilm shrinkage in the cellular automaton method. The new pore-scale biofilm model is then evaluated against a previously published pore-scale biofilm experiment, in which two microfluidic flow cells, one with a homogeneous pore network and the other with an aggregate pore network, were tested for aerobic degradation of a herbicide. The simulated biofilm distribution and morphology, biomass accumulation, and contaminant removal are generally consistent with the experimental data. Biofilm detachment in this model occurs when the local shear stress is above a critical value. We use the critical value from our previously published modeling study and find it works well in this case, even though we now have a different pore network and a different microbial species. We also use the model to show that the interaction between flow and biofilm growth is important to predict contaminant removal. The computational time of the new model is reduced 90% compared to our prior work due to implementation of biofilm shrinkage in the cellular automaton method. To the best of our knowledge, this is the first time that biofilm shrinkage has been incorporated into a pore-scale model for simulation of pollutant biodegradation in porous media.

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For the first time, we demonstrate chromate (Cr(VI)) bioreduction using methane (CH4) as the sole electron donor in a membrane biofilm reactor (MBfR). The experiments were divided into five stages lasting a total of 90 days, and each stage achieved a steady state for at least 15 days. Due to continued acclimation of the microbial community, the Cr(VI)-reducing capacity of the biofilm kept increasing. Cr(VI) removal at the end of the 90-day test reached 95% at an influent Cr(VI) concentration of 3 mg Cr/L and a surface loading of 0.37g of Cr m(-2) day(-1). Meiothermus (Deinococci), a potential Cr(VI)-reducing bacterium, was negligible in the inoculum but dominated the MBfR biofilm after Cr(VI) was added to the reactor, while Methylosinus, a type II methanotrophs, represented 11%-21% of the total bacterial DNA in the biofilm. Synergy within a microbial consortia likely was responsible for Cr(VI) reduction based on CH4 oxidation. In the synergy, methanotrophs fermented CH4 to produce metabolic intermediates that were used by the Cr(VI)-reducing bacteria as electron donors. Solid Cr(III) was the main product, accounting for more than 88% of the reduced Cr in most cases. Transmission electron microscope (TEM) and energy dispersive X-ray (EDS) analysis showed that Cr(III) accumulated inside and outside of some bacterial cells, implying that different Cr(VI)-reducing mechanisms were involved.

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To study the effect of nitrate (NO3(-)) on selenate (SeO4(2-)) reduction, we tested a H2-based biofilm with a range of influent NO3(-) loadings. When SeO4(2-) was the only electron acceptor (stage 1), 40% of the influent SeO4(2-) was reduced to insoluble elemental selenium (Se(0)). SeO4(2-) reduction was dramatically inhibited when NO3(-) was added at a surface loading larger than 1.14 g of N m(-2) day(-1), when H2 delivery became limiting and only 80% of the input NO3(-) was reduced (stage 2). In stage 3, when NO3(-) was again removed from the influent, SeO4(2-) reduction was re-established and increased to 60% conversion to Se(0). SeO4(2-) reduction remained stable at 60% in stages 4 and 5, when the NO3(-) surface loading was re-introduced at ≤ 0.53 g of N m(-2) day(-1), allowing for complete NO3(-) reduction. The selenate-reducing microbial community was significantly reshaped by the high NO3(-) surface loading in stage 2, and it remained stable through stages 3-5. In particular, the abundance of α-Proteobacteria decreased from 30% in stage 1 to less than 10% of total bacteria in stage 2. β-Proteobacteria, which represented about 55% of total bacteria in the biofilm in stage 1, increased to more than 90% of phylotypes in stage 2. Hydrogenophaga, an autotrophic denitrifier, was positively correlated with NO3(-) flux. Thus, introducing a NO3(-) loading high enough to cause H2 limitation and suppress SeO4(2-) reduction had a long-lasting effect on the microbial community structure, which was confirmed by principal coordinate analysis, although SeO4(2-) reduction remained intact.

Selenate (SeO4(2-)) bioreduction is possible with oxidation of a range of organic or inorganic electron donors, but it never has been reported with methane gas (CH4) as the electron donor. In this study, we achieved complete SeO4(2-) bioreduction in a membrane biofilm reactor (MBfR) using CH4 as the sole added electron donor. The introduction of nitrate (NO3(-)) slightly inhibited SeO4(2-) reduction, but the two oxyanions were simultaneously reduced, even when the supply rate of CH4 was limited. The main SeO4(2-)-reduction product was nanospherical Se(0), which was identified by scanning electron microscopy coupled to energy dispersive X-ray analysis (SEM-EDS). Community analysis provided evidence for two mechanisms for SeO4(2-) bioreduction in the CH4-based MBfR: a single methanotrophic genus, such as Methylomonas, performed CH4 oxidation directly coupled to SeO4(2-) reduction, and a methanotroph oxidized CH4 to form organic metabolites that were electron donors for a synergistic SeO4(2-)-reducing bacterium.

This work presents a multispecies biofilm model that describes the co-existence of nitrate- and sulfate-reducing bacteria in the H(2)-based membrane biofilm reactor (MBfR). The new model adapts the framework of a biofilm model for simultaneous nitrate and perchlorate removal by considering the unique metabolic and physiological characteristics of autotrophic sulfate-reducing bacteria that use H(2) as their electron donor. To evaluate the model, the simulated effluent H(2), UAP (substrate-utilization-associated products), and BAP (biomass-associated products) concentrations are compared to experimental results, and the simulated biomass distributions are compared to real-time quantitative polymerase chain reaction (qPCR) data in the experiments for parameter optimization. Model outputs and experimental results match for all major trends and explain when sulfate reduction does or does not occur in parallel with denitrification. The onset of sulfate reduction occurs only when the nitrate concentration at the fiber's outer surface is low enough so that the growth rate of the denitrifying bacteria is equal to that of the sulfate-reducing bacteria. An example shows how to use the model to design an MBfR that achieves satisfactory nitrate reduction, but suppresses sulfate reduction.