Concomitant urea stabilization and phosphorus recovery from source-separated fresh urine in magnesium anode-based peroxide-producing electrochemical cells.

In this study, we investigated the recovery of nitrogen (N) and phosphorus (P) from fresh source-separated urine with a novel electrochemical cell equipped with a magnesium (Mg) anode and carbon-based gas-diffusion cathode. Recovery of P, which exists primarily as phosphate (PO43-) in urine, was achieved through pH-driven precipitation. Maximizing N recovery requires simultaneous approaches to address urea and ammonia (NH3). NH3 recovery was possible through precipitation in struvite with soluble Mg supplied by the anode. Urea was stabilized with electrochemically synthesized hydrogen peroxide (H2O2) from the cathode. H2O2 concentrations and resulting urine pH were directly proportional to the applied current density. Concomitant NH3 and PO43- precipitation as struvite and urea stabilization via H2O2 electrosynthesis was possible at lower current densities, resulting in urine pH under 9.2. Higher current densities resulted in urine pH over 9.2, yielding higher H2O2 concentrations and more consistent stabilization of urea at the expense of NH3 recovery as struvite; PO43- precipitation still occurred but in the form of calcium phosphate and magnesium phosphate solids.

Predicting COVID-19 Infected Individuals in a Defined Population from Wastewater RNA Data.

Wastewater surveillance of SARS-CoV-2 RNA has become an important tool for tracking the presence of the virus and serving as an early indicator for the onset of rapid transmission. Nevertheless, wastewater data are still not commonly used to predict the number of infected individuals in a sewershed. The main objective of this study was to calibrate a susceptible-exposed-infectious-recovered (SEIR) model using RNA copy rates in sewage (i.e., gene copies per liter times flow rate) and the number of SARS-CoV-2 saliva-test-positive infected individuals in a university student population that was subject to repeated weekly testing during the Spring 2021 semester. A strong correlation was observed between the RNA copy rates and the number of infected individuals. The parameter in the SEIR model that had the largest impact on calibration was the maximum shedding rate, resulting in a mean value of 7.72 log10 genome copies per gram of feces. Regressing the saliva-test-positive infected individuals on predictions from the SEIR model based on the RNA copy rates yielded a slope of 0.87 (SE=0.11), which is statistically consistent with a 1:1 relationship between the two. These findings demonstrate that wastewater surveillance of SARS-CoV-2 can be used to estimate the number of infected individuals in a sewershed.

COVID-19 wastewater epidemiology: a model to estimate infected populations.

BACKGROUND: Wastewater-based epidemiology provides an opportunity for near real-time, cost-effective monitoring of community-level transmission of SARS-CoV-2. Detection of SARS-CoV-2 RNA in wastewater can identify the presence of COVID-19 in the community, but methods for estimating the numbers of infected individuals on the basis of wastewater RNA concentrations are inadequate. METHODS: This is a wastewater-based epidemiology study using wastewater samples that were collected weekly or twice a week from three sewersheds in South Carolina, USA, between either May 27 or June 16, 2020, and Aug 25, 2020, and tested for SARS-CoV-2 RNA. We developed a susceptible-exposed-infectious-recovered (SEIR) model based on the mass rate of SARS-CoV-2 RNA in the wastewater to predict the number of infected individuals, and have also provided a simplified equation to predict this. Model predictions were compared with the number of confirmed cases identified by the Department of Health and Environmental Control, South Carolina, USA, for the same time period and geographical area. FINDINGS: We plotted the model predictions for the relationship between mass rate of virus release and numbers of infected individuals, and we validated this prediction on the basis of estimated prevalence from individual testing. A simplified equation to estimate the number of infected individuals fell within the 95% confidence limits of the model. The rate of unreported COVID-19 cases, as estimated by the model, was approximately 11 times that of confirmed cases (ie, ratio of estimated infections for every confirmed case of 10·9, 95% CI 4·2-17·5). This rate aligned well with an independent estimate of 15 infections for every confirmed case in the US state of South Carolina. INTERPRETATION: The SEIR model provides a robust method to estimate the total number of infected individuals in a sewershed on the basis of the mass rate of RNA copies released per day. This approach overcomes some of the limitations associated with individual testing campaigns and thereby provides an additional tool that can be used to inform policy decisions. FUNDING: Clemson University, USA.

Evaluation of electrical current production in microbial electrolysis cells fed with animal rendering wastewater.

Anode-respiring bacteria (ARB) generate electrical current from the oxidation of short chain fatty acids (SCFA), primarily acetate, in microbial electrolysis cells (MECs). Animal rendering wastewater (RW) has high fat content, which under anaerobic conditions can yield acetate, making RW a potential feed for MECs. Yet, excess intermediate long chain fatty acids (LCFA) may limit conversion of LCFA and SCFA, and impact ARB activity. Here, we evaluated electrical current production in single-chamber MECs fed with RW. In RW-fed MECs, 34.26 ± 2.69% of the COD provided was converted to electrical current in an 80-day batch cycle. LCFA accumulated in RW-fed MECs, during which conversion of acetate to electrical current was limited. Diverse sulfate-reducing microorganisms were present in the anode biofilm in RW-fed MECs, whereas the genus Geobacter dominated in inoculum-only control MECs. Detection of H2-utilizing homoacetogens suggested some internal cycling of H2 produced at the cathode. Overall, this study shows that current production is possible from RW, but to be a viable process for RW treatment, further improvement in rates of COD conversion and current production is necessary along with identifying configurations and/or conditions in which the inhibitory effect of LCFA is reduced.

Water Recovery from Bioreactor Mixed Liquors Using Forward Osmosis with Polyelectrolyte Draw Solutions.

This paper reports on the use of forward osmosis (FO) with polyelectrolyte draw solutions to recover water from bioreactor mixed liquors. The work was motivated by the need for new regenerative water purification technologies to enable long-duration space missions. Osmotic membrane bioreactors may be an option for water and nutrient recovery in space if they can attain high water flux and reverse solute flux selectivity (RSFS), which quantifies the mass of permeated water per mass of draw solute that has diffused from the draw solution into a bioreactor. Water flux was measured in a direct flow system using wastewater from a municipal wastewater treatment plant and draw solutions prepared with two polyelectrolytes at different concentrations. The direct flow tests displayed a high initial flux (>10 L/m2/h) that decreased rapidly as solids accumulated on the feed side of the membrane. A test with deionized water as the feed revealed a small mass of polyelectrolyte crossover from the draw solution to the feed, yielding an RSFS of 80. Crossflow filtration experiments demonstrated that steady state flux above 2 L/m2·h could be maintained for 70 h following an initial flux decline due to the formation of a foulant cake layer. This study established that FO could be feasible for regenerative water purification from bioreactors. By utilizing a polyelectrolyte draw solute with high RSFS, we expect to overcome the need for draw solute replenishment. This would be a major step towards sustainable operation in long-duration space missions.

pH Dependency in Anode Biofilms of Thermincola ferriacetica Suggests a Proton-Dependent Electrochemical Response.

Monitoring the electrochemical response of anode respiring bacteria (ARB) helps elucidate the fundamental processes of anode respiration and their rate limitations. Understanding these limitations provides insights on how ARB create the complex interfacing of biochemical metabolic processes with insoluble electron acceptors and electronics. In this study, anode biofilms of the thermophilic (60 °C) Gram-positive ARB Thermincola ferriacetica were studied to determine the presence of a proton-dependent electron transfer response. The effects of pH, the presence of an electron donor (acetate), and biofilm growth were varied to determine their influence on the electrochemical midpoint potential ( EKA) and formal redox potential ( E°') under nonturnover conditions. The EKA and E°' are associated with an enzymatic process within ARB's metabolism that controls the rate and energetic state of their respiration. Results for all conditions indicate that pH was the major contributor to altering the energetics of T. ferriacetica anode biofilms. Electrochemical responses measured in the absence of an electron donor and with a minimal proton gradient within the anode biofilms resulted in a 48 ± 7 mV/pH unit shift in the E°', suggesting a proton-dependent rate-limiting process. Given the limited energy available for anode respiration (<200 mV when using acetate as electron donor), our results provide a new perspective in understanding proton-transport limitations in ARB biofilms, one in which ARB are thermodynamically limited by pH gradients. Since the anode biofilms of all ARB that perform direct extracellular electron transfer (EET) investigated thus far exhibit an n = 1 Nernstian behavior, and because this behavior is affected by changes in pH, we hypothesize that the Nernstian response is associated with membrane proteins responsible for proton translocation. Finally, this study shows that the EKA and E°' are a function of pH within the physiological range of ARB, and thus, given the significant effect pH has on this parameter, we recommend reporting the EKA and E°' of ARB biofilms at a specific bulk pH.

H2O2 Production in Microbial Electrochemical Cells Fed with Primary Sludge.

We developed an energy-efficient, flat-plate, dual-chambered microbial peroxide producing cell (MPPC) as an anaerobic energy-conversion technology for converting primary sludge (PS) at the anode and producing hydrogen peroxide (H2O2) at the cathode. We operated the MPPC with a 9 day hydraulic retention time in the anode. A maximum H2O2 concentration of ∼230 mg/L was achieved in 6 h of batch cathode operation. This is the first demonstration of H2O2 production using PS in an MPPC, and the energy requirement for H2O2 production was low (∼0.87 kWh/kg H2O2) compared to previous studies using real wastewaters. The H2O2 gradually decayed with time due to the diffusion of H2O2-scavenging carbonate ions from the anode. We compared the anodic performance with a H2-producing microbial electrolysis cell (MEC). Both cells (MEC and MPPC) achieved ∼30% Coulombic recovery. While similar microbial communities were present in the anode suspension and anode biofilm for the two operating modes, aerobic bacteria were significant only on the side of the anode facing the membrane in the MPPC. Coupled with a lack of methane production in the MPPC, the presence of aerobic bacteria suggests that H2O2 diffusion to the anode side caused inhibition of methanogens, which led to the decrease in chemical oxygen demand removal. Thus, the Coulombic efficiency was ∼16% higher in the MPPC than in the MEC (64% versus 48%, respectively).

Tailoring Microbial Electrochemical Cells for Production of Hydrogen Peroxide at High Concentrations and Efficiencies.

A microbial peroxide producing cell (MPPC) for H2 O2 production at the cathode was systematically optimized with minimal energy input. First, the stability of H2 O2 was evaluated using different catholytes, membranes, and catalyst materials. On the basis of these results, a flat-plate MPPC fed continuously using 200 mm NaCl catholyte at a 4 h hydraulic retention time was designed and operated, producing H2 O2 for 18 days. H2 O2 concentration of 3.1 g L-1 H2 O2 with 1.1 Wh g-1 H2 O2 power input was achieved in the MPPC. The high H2 O2 concentration was a result of the optimum materials selected. The small energy input was largely the result of the 0.5 cm distance between the anode and cathode, which reduced ionic transport losses. However, >50 % of operational overpotentials were due to the 4.5-5 pH unit difference between the anode and cathode chambers. The results demonstrate that a MPPC can continuously produce H2 O2 at high concentration by selecting compatible materials and appropriate operating conditions.

The effect of pH and buffer concentration on anode biofilms of Thermincola ferriacetica.

We assessed the effects of pH and buffer concentration on current production and growth of biofilms of Thermincola ferriacetica - a thermophilic, Gram-positive, anode-respiring bacterium (ARB) - grown on anodes poised at a potential of -0.06V vs. SHE in microbial electrolysis cells (MECs) at 60°C. T. ferriacetica generated current in the pH range of 5.2 to 8.3 with acetate as the electron donor and 50mM bicarbonate buffer. Maximum current density was reduced by ~80% at pH5.2 and ~14% at 7.0 compared to pH8.3. Increasing bicarbonate buffer concentrations from 10mM to 100mM resulted in an increase in the current density by 40±6%, from 6.8±1.1 to 11.2±2.7Am(-2), supporting that more buffer alleviated pH depression within T. ferriacetica biofilms. Confocal laser scanning microscopy (CLSM) images indicated that higher bicarbonate buffer concentrations resulted in larger live biofilm thicknesses: from 68±20µm at 10mM bicarbonate to >150µm at 100mM, supporting that buffer availability was a strong influence on biofilm thickness. In comparison to mesophilic Geobacter sulfurreducens biofilms, the faster transport rates at higher temperature and the ability to grow at relatively lower pH allowed T. ferriacetica to produce higher current densities with lower buffer concentrations.

Critical transport rates that limit the performance of microbial electrochemistry technologies.

Microbial electrochemistry technologies (METs) take advantage of the connection of microorganisms with electrodes. In the classic case of a microbial anode, the maximization of current density produced is often the goal. But, current production is dependent on many transport processes occurring, which can be rate-limiting. These include the fluxes of electron donor and acceptor, the ionic flux, the acidity and alkalinity fluxes at anode and cathode respectively, the electron transport flux at the biofilm, and the reactant/product crossover flux. Associated with these fluxes are inherent concentration gradients that can affect performance. This critical review provides an analysis on how these transport processes have hindered the development of METs, and how MET designs have evolved as more knowledge of these transport limitations is gained. Finally, suggestions are provided on how to design MET systems taking into consideration critical transport processes that are intimately linked to the current produced.

Application of microbial electrolysis cells to treat spent yeast from an alcoholic fermentation.

Spent yeast (SY), a major challenge for the brewing industry, was treated using a microbial electrolysis cell to recover energy. Concentrations of SY from bench alcoholic fermentation and ethanol were tested, ranging from 750 to 1500mgCOD/L and 0 to 2400mgCOD/L respectively. COD removal efficiency (RE), coulombic efficiency (CE), coulombic recovery (CR), hydrogen production and current density were evaluated. The best treatment condition was 750mgCOD/LSY+1200mgCOD/L ethanol giving higher COD RE, CE, CR (90±1%, 90±2% and 81±1% respectively), as compared with 1500mgCOD/LSY (76±2%, 63±7% and 48±4% respectively); ethanol addition was significantly favorable (p value=0.011), possibly due to electron availability and SY autolysis. 1500mgCOD/LSY+1200mgCOD/L ethanol achieved higher current density (222.0±31.3A/m(3)) and hydrogen production (2.18±0.66 [Formula: see text] ) but with lower efficiencies (87±2% COD RE, 71.0±.4% CE). Future work should focus on electron sinks, acclimation and optimizing SY breakdown.

Anode Biofilms of Geoalkalibacter ferrihydriticus Exhibit Electrochemical Signatures of Multiple Electron Transport Pathways.

Thriving under alkaliphilic conditions, Geoalkalibacter ferrihydriticus (Glk. ferrihydriticus) provides new applications in treating alkaline waste streams as well as a possible new model organism for microbial electrochemistry. We investigated the electrochemical response of biofilms of the alkaliphilic anode-respiring bacterium (ARB) Glk. ferrihydriticus voltammetry (CV), electrochemical impedance spectroscopy (EIS), and chronoamperometry. We observed there to be at least four dominant electron transfer pathways, with their contribution to the overall current produced dependent on the set anode potential. These pathways appear to be manifested at midpoint potentials of approximately -0.14 V, -0.2 V, -0.24 V, and -0.27 V vs standard hydrogen electrode. The individual contributions of the pathways change upon equilibration from a set anode potential to another anode potential. Additionally, the contribution of each pathway to the overall current produced is reversible when the anode potential is changed back to the original set potential. The pathways involved in anode respiration in Glk. ferrihydriticus biofilms follow a similar, but more complicated, pattern as compared to those in the model ARB, Geobacter sulfurreducens. This greater diversity of electron transport pathways in Glk. ferrihydriticus could be related to its wider metabolic capability (e.g., higher pH and larger set of possible substrates, among others).

Effects of pre-fermentation and pulsed-electric-field treatment of primary sludge in microbial electrochemical cells.

The aim of this study was to investigate the combination of two technologies - pulsed electric field (PEF) pre-treatment and semi-continuous pre-fermentation of primary sludge (PS) - to produce volatile fatty acids (VFAs) as the electron donor for microbial electrolysis cells (MECs). Pre-fermentation with a 3-day solids retention time (SRT) led to the maximum generation of VFAs, with or without pretreatment of the PS through pulsed-electric-fields (PEF). PEF treatment before fermentation enhanced the accumulation of the preferred VFA, acetate, by 2.6-fold. Correspondingly, MEC anodes fed with centrate from 3-day pre-fermentation of PEF-treated PS had a maximum current density ∼3.1 A/m(2), which was 2.4-fold greater than the control pre-fermented centrate. Over the full duration of batch MEC experiments, using pre-fermented centrate led to successful performance in terms of Coulombic efficiency (95%), Coulombic recovery (80%), and COD-removal efficiency (85%).

Dynamic potential-dependent electron transport pathway shifts in anode biofilms of Geobacter sulfurreducens.

Biofilms of the anode-respiring bacterium Geobacter sulfurreducens (G. sulfurreducens) demonstrate dynamic potential-dependent changes between two electron transport pathways that are used selectively depending on the anode potential. Electrochemical impedance spectroscopy (EIS) measurements suggest that these pathways (both n=1), with midpoint potentials of -0.155 (± 0.005) and -0.095 (± 0.003) V versus standard hydrogen electrode, are not additive within the biofilm, but are preferentially used depending on the anode potential. Potential step voltammetry and cyclic voltammetry (CV) confirmed rapid changes between the two pathways in minutes when the anode potential is changed. We confirm that the electrochemical response observed in a slow-scan-rate CV (∼1 mV s(-1) ) is often composed of at least the two pathways characterized. Thus, beyond understanding the electron transport pathways in G. sulfurreducens, this study also has implications on the interpretation of previously collected and future potential-dependent datasets.

Selective enrichment yields robust ethene-producing dechlorinating cultures from microcosms stalled at cis-dichloroethene.

Dehalococcoides mccartyi strains are of particular importance for bioremediation due to their unique capability of transforming perchloroethene (PCE) and trichloroethene (TCE) to non-toxic ethene, through the intermediates cis-dichloroethene (cis-DCE) and vinyl chloride (VC). Despite the widespread environmental distribution of Dehalococcoides, biostimulation sometimes fails to promote dechlorination beyond cis-DCE. In our study, microcosms established with garden soil and mangrove sediment also stalled at cis-DCE, albeit Dehalococcoides mccartyi containing the reductive dehalogenase genes tceA, vcrA and bvcA were detected in the soil/sediment inocula. Reductive dechlorination was not promoted beyond cis-DCE, even after multiple biostimulation events with fermentable substrates and a lengthy incubation. However, transfers from microcosms stalled at cis-DCE yielded dechlorination to ethene with subsequent enrichment cultures containing up to 10(9) Dehalococcoides mccartyi cells mL(-1). Proteobacterial classes which dominated the soil/sediment communities became undetectable in the enrichments, and methanogenic activity drastically decreased after the transfers. We hypothesized that biostimulation of Dehalococcoides in the cis-DCE-stalled microcosms was impeded by other microbes present at higher abundances than Dehalococcoides and utilizing terminal electron acceptors from the soil/sediment, hence, outcompeting Dehalococcoides for H2. In support of this hypothesis, we show that garden soil and mangrove sediment microcosms bioaugmented with their respective cultures containing Dehalococcoides in high abundance were able to compete for H2 for reductive dechlorination from one biostimulation event and produced ethene with no obvious stall. Overall, our results provide an alternate explanation to consolidate conflicting observations on the ubiquity of Dehalococcoides mccartyi and occasional stalling of dechlorination at cis-DCE; thus, bringing a new perspective to better assess biological potential of different environments and to understand microbial interactions governing bioremediation.

Successful operation of continuous reactors at short retention times results in high-density, fast-rate Dehalococcoides dechlorinating cultures.

The discovery of Dehalococcoides mccartyi reducing perchloroethene and trichloroethene (TCE) to ethene was a key landmark for bioremediation applications at contaminated sites. D. mccartyi-containing cultures are typically grown in batch-fed reactors. On the other hand, continuous cultivation of these microorganisms has been described only at long hydraulic retention times (HRTs). We report the cultivation of a representative D. mccartyi-containing culture in continuous stirred-tank reactors (CSTRs) at a short, 3-d HRT, using TCE as the electron acceptor. We successfully operated 3-d HRT CSTRs for up to 120 days and observed sustained dechlorination of TCE at influent concentrations of 1 and 2 mM TCE to ≥ 97 % ethene, coupled to the production of 10(12) D. mccartyi cells Lculture (-1). These outcomes were possible in part by using a medium with low bicarbonate concentrations (5 mM) to minimize the excessive proliferation of microorganisms that use bicarbonate as an electron acceptor and compete with D. mccartyi for H2. The maximum conversion rates for the CSTR-produced culture were 0.13 ± 0.016, 0.06 ± 0.018, and 0.02 ± 0.007 mmol Cl(-) Lculture (-1) h(-1), respectively, for TCE, cis-dichloroethene, and vinyl chloride. The CSTR operation described here provides the fastest laboratory cultivation rate of high-cell density Dehalococcoides cultures reported in the literature to date. This cultivation method provides a fundamental scientific platform for potential future operations of such a system at larger scales.

Kinetic, electrochemical, and microscopic characterization of the thermophilic, anode-respiring bacterium Thermincola ferriacetica.

Thermincola ferriacetica is a recently isolated thermophilic, dissimilatory Fe(III)-reducing, Gram-positive bacterium with capability to generate electrical current via anode respiration. Our goals were to determine the maximum rates of anode respiration by T. ferriacetica and to perform a detailed microscopic and electrochemical characterization of the biofilm anode. T. ferriacetica DSM 14005 was grown at 60 °C on graphite-rod anodes poised at -0.06 V (vs) SHE in duplicate microbial electrolysis cells (MECs). The cultures grew rapidly until they achieved a sustained current density of 7-8 A m(-2) with only 10 mM bicarbonate buffer and an average Coulombic Efficiency (CE) of 93%. Cyclic voltammetry performed at maximum current density revealed a Nernst-Monod response with a half saturation potential (EKA) of -0.127 V (vs) SHE. Confocal microscopy images revealed a thick layer of actively respiring cells of T. ferriacetica (~38 µm), which is the first documentation for a gram positive anode respiring bacterium (ARB). Scanning electron microscopy showed a well-developed biofilm with a very dense network of extracellular appendages similar to Geobacter biofilms. The high current densities, a thick biofilm (~38 µm) with multiple layers of active cells, and Nernst-Monod behavior support extracellular electron transfer (EET) through a solid conductive matrix - the first such observation for Gram-positive bacteria. Operating with a controlled anode potential enabled us to grow T. ferriacetica that can use a solid conductive matrix resulting in high current densities that are promising for MXC applications.

Importance of OH(-) transport from cathodes in microbial fuel cells.

Cathodic limitation in microbial fuel cells (MFCs) is considered an important hurdle towards practical application as a bioenergy technology. The oxygen reduction reaction (ORR) needs to occur in MFCs under significantly different conditions compared to chemical fuel cells, including a neutral pH. The common reason cited for cathodic limitation is the difficulty in providing protons to the catalyst sites. Here, we show that it is not the availability of protons, but the transport of OH(-) from the catalyst layer to the bulk liquid that largely governs cathodic potential losses. OH(-) is a product of an ORR mechanism that has not been considered dominant before. The accumulation of OH(-) at the catalyst sites results in an increase in the local cathode pH, resulting in Nernstian concentration losses. For Pt-based gas-diffusion cathodes, using polarization curves developed in unbuffered and buffered solutions, we quantified this loss to be >0.3 V at a current density of 10 Am(-2) . We show that this loss can be partially overcome by replacing the Nafion binder used in the cathode catalyst layer with an anion-conducting binder and by providing additional buffer to the cathode catalyst directly in the form of CO(2) , which results in enhanced OH(-) transport. Our results provide a comprehensive analysis of cathodic limitations in MFCs and should allow researchers to develop and select materials for the construction of MFC cathodes and identify operational conditions that will help minimize Nernstian concentration losses due to pH gradients.

Using electron balances and molecular techniques to assess trichoroethene-induced shifts to a dechlorinating microbial community.

This study demonstrated the utility in correlating performance and community structure of a trichloroethene (TCE)-dechlorinating microbial consortium; specifically dechlorinators, fermenters, homoacetogens, and methanogens. Two complementary approaches were applied: predicting trends in the microbial community structure based on an electron balance analysis and experimentally assessing the community structure via pyrosequencing and quantitative polymerase chain reaction (qPCR). Fill-and-draw reactors inoculated with the DehaloR^2 consortium were operated at five TCE-pulsing rates between 14 and 168 µmol/10-day-SRT, amended with TCE every 2 days to give peak concentrations between 0.047 and 0.56 mM (6-74 ppm) and supplied lactate and methanol as sources of e(-) donor and carbon. The complementary approaches demonstrated the same trends: increasing abundance of Dehalococcoides and Geobacter and decreasing abundance of Firmicutes with increasing TCE pulsing rate, except for the highest pulsing rate. Based on qPCR, the abundance of Geobacter and Dehalococcoides decreased for the highest TCE pulsing rate, and pyrosequencing showed this same trend for the latter. This deviation suggested decoupling of Dehalococcoides growth from dechlorination. At pseudo steady-state, methanogenesis was minimal for all TCE pulsing rates. Pyrosequencing and qPCR showed suppression of the homoacetogenic genera Acetobacterium at the two highest pulsing rates, and it was corroborated by a decreased production of acetate from lactate fermentation and increased propionate production. Suppression of Acetobacterium, which can provide growth factors to Dehalococcoides, may have contributed to the decoupling for the highest TCE-pulsing rate.

Managing methanogens and homoacetogens to promote reductive dechlorination of trichloroethene with direct delivery of H2 in a membrane biofilm reactor.

A study with H(2)-based membrane biofilm reactors (MBfRs) was undertaken to examine the effectiveness of direct H(2) delivery in ex-situ reductive dechlorination of chlorinated ethenes. Trichloroethene (TCE) could be reductively dechlorinated to ethene with up to 95% efficiency as long as the pH-increase effects of methanogens and homoacetogens were managed and dechlorinators were selected for during start-up by creating H(2) limitation. Based on quantitative PCR, the dominant bacterial groups in the biofilm at the end of reactor operation were Dehalococcoides, Geobacter, and homoacetogens. Pyrosequencing confirmed the dominance of the dechlorinators and identified Acetobacterium as the key homoacetogen. Homoacetogens outcompeted methanogens for bicarbonate, based on the effluent concentration of acetate, by suppressing methanogens during batch start-up. This was corroborated by the methanogenesis functional gene mcrA, which was 1-2 orders of magnitude lower than the FTHFS functional gene for homoacetogens. Imaging of the MBfR fibers using scanning electron microscopy showed a distinct Dehalococcoides-like morphology in the fiber biofilm. These results support that direct addition of H(2) can allow for efficient and complete reductive dechlorination, and they shed light into how H(2)-fed biofilms, when operated to manage methanogenic and homoacetogenic activity, can be used for ex-situ bioremediation of chlorinated ethenes.