A Monod-based model of attached-growth anaerobic fermenters

A simple mathematical model of steady-state attached-growth anaerobic fermenter kinetics is described. The model considers a single methanogenic culture following Monod growth kinetics. The model accounts for effects of influent biodegradability and volatile solids concentration, temperature and hydraulic retention time, and predicts volumetric methane productivity and volatile solids reduction. It is distinguished from conventional suspended-growth reactor models by its explicit consideration of bacterial concentration in the reactor system, based on hydraulic flow and influent volatile solids concentration. The model was validated using data from both porous- and solid-media attached-growth fermenters.     

Treatment of pig manure liquid digestate in horizontal flow constructed wetlands: Effect of aeration

With the rapid development of scaled anaerobic digestion of pig manure, the generation of liquid anaerobic digestate exceeds the farmland loading capacity, causing serious environmental pollution. Three laboratory‐scale horizontal subsurface flow constructed wetlands (CWs; planted + aeration, planted, and unplanted) were set up to investigate the feasibility of liquid digestate treatment in wetlands. Treatment capacity in different wetlands was evaluated under different influent concentrations (chemical oxygen demand [COD], 5 days biochemical oxygen demand [BOD₅], and nitrogen forms). The effect of aeration and effluent recirculation on organic matter and total nitrogen removal was investigated. Results showed that integrating intermittent aeration in CWs significantly improved the oxygen condition (p < 0.01) in the wetland bed and promoted BOD₅ removal to 90% in aerated CWs as compared with <15% in the unaerated CWs. Meanwhile, COD removal between these three wetlands did not show any difference and varied from 52 to 72% under influent concentration of 200–820 mg/L because of the high content of hard‐degradable organic matter in the liquid digestate. Intermittent aeration resulted in high ammonium removal (>98%) although the influent loading varied from 65 to 350 mg/L. However, intermittent aeration caused nitrate accumulation of 300 mg/L and limited total nitrogen (TN) removal of 33%. To intensify the TN removal, we verified effluent recirculation to increase the removal efficiency of TN to 78%. These results not only show the potential application of CWs for treatment of high‐strength liquid anaerobic digested slurry, but also indicate the significance of intermittent aeration on the enhanced removal of organic matter and ammonium.     

Anaerobic treatment of low-strength municipal wastewater by a two-stage pilot plant under psychrophilic conditions

A two-stage anaerobic treatment pilot plant was tested for the treatment of raw domestic wastewater under temperatures ranging from 21 to 14 degrees C. The plant consisted of a hydrolytic upflow sludge bed (HUSB) digester (25.5m3) followed by an upflow anaerobic sludge blanket (UASB) digester (20.36m3). The hydraulic retention time (HRT) varied from 5.7 to 2.8h for the first stage (HUSB digester) and from 13.9 to 6.5h for the second stage (UASB digester). Total suspended solids (TSS), total chemical oxygen demand (TCOD), and biochemical oxygen demand (BOD) removals ranged from 76% to 89%, from 49% to 65%, and from 50% to 77%, respectively, for the overall system. The percentage of influent COD converted to methane was 36.1%, the hydrolysis of influent volatile suspended solids (VSS) reached 59.7% and excess biomass was 21.6% of the incoming VSS. Plant performance was influenced by the wastewater concentration and temperature, yet better results were obtained for influent COD higher than 250mg/l.     

Understanding the Anodic Mechanism of a Seafloor Fuel Cell: Interactions Between Geochemistry and Microbial Activity

Seafloor fuel cells made with graphite electrodes generate electricity by promoting electron transfer in response to a natural voltage difference (−0.7 to −0.8 V) between anoxic sediments and overlying oxic seawater. Geochemical impacts of a seafloor fuel cell on sediment solids and porewaters were examined to identify the anodic mechanisms and substrates available for current production. In an estuarine environment with little dissolved sulfide, solid-phase acid volatile sulfide and Cr²⁺-reducible sulfur minerals decreased significantly toward the anode after 7 months of nearly continuous energy harvesting. Porewater iron and sulfate increased by millimolar amounts. Scanning electron microscope images showed a biofilm overcoating the anode, and electron microprobe analyses revealed accumulations of sulfur, iron, silicon and phosphorus at the electrode surface. Sulfur deposition was also observed on a laboratory fuel cell anode used to generate electricity with only dissolved sulfide as an electron donor. Moreover, current densities and voltages displayed by these purely chemical cells were similar to the values measured with field devices. These results indicate that electron transfer to seafloor fuel cells can readily result in the oxidation of dissolved and solid-phase forms of reduced sulfur producing mainly S⁰ which deposits at the electrode surface. This oxidation product is consistent with the observed enrichment of bacteria most closely related to Desulfobulbus/Desulfocapsa genera within the anode biofilm, and its presence is proposed to promote a localized biogeochemical cycle whereby biofilm bacteria regenerate sulfate and sulfide. This electron-shuttling mechanism may co-occur while these or other bacteria use the anode directly as a terminal electron acceptor.     

Analysis of the performance of an anaerobic digestion system at the Regina Wastewater Treatment Plant

From the performance analysis of the anaerobic digestion system at the Regina Wastewater Treatment Plant, it was found that the anaerobic digestion system at the Regina plant was generally operated in a stable condition as indicated by pH, volatile acids and alkalinity levels. The operation of the anaerobic digestion system was not optimal because of the low volatile solids concentration and low volatile solids loading rate, especially because of high HRT. Two options, thickening the primary sludge and increasing the volatile solids loading rate, were recommended for the optimal operation of the digestion system. After examining a number of kinetic models, it was found that the Chen-Hashimoto model could be used to predict the volumetric methane production rate and the first-order model could be used to predict the efficiency of volatile solids reduction. The study showed that utilization of digester gas for power production was the best alternative for the excess digester gas. 13.3% of the electrical demand and 35.5% of the plant's total energy could be met based on digester gas wasted, assuming 25% as the conversion efficiency.     

Effect of effluent recycling and shock loading on the biodegradation of complex phenolic mixture in hybrid UASB reactors

This study describes the feasibility of anaerobic treatment of synthetic coal wastewater using four identical 13.5L (effective volume) bench scale hybrid up flow anaerobic sludge blanket (HUASB) reactors (R1, R2, R3 and R4) under mesophilic (27+/-5 degrees C) conditions. Synthetic coal wastewater with an average chemical oxygen demand (COD) of 2240 mg/L and phenolics concentration of 752 mg/L was used as substrate. Effluent recirculation was employed at four different effluent to feed recirculation ratios (R/F) of 0.5, 1.0, 1.5 and 2.0 for 100 days to study the effect of recirculation on the performance of the reactors. Phenolics and COD removal was found to improve with increase in effluent recirculation. An effluent to feed recycle ratio of 1.0 resulted in maximum removal of phenolics and COD. Phenolics and COD removal improved from 88% and 92% to 95% each, respectively. The concentration of volatile fatty acids in the effluent was lower than the influent when effluent to feed recirculation was employed. Effect of shock loading on the reactors revealed that phenolics shock load up to 2.5 times increase in the normal input phenolics concentration in the form of continuous shock load for 4days did not affect the reactors performance irreversibly.     

Thermal Energy Interchange During Anaerobic Methane Fermentation of Waste Organic Substrates

An experimental calorimeter-type anaerobic fermentation system was developed to evaluate the extent of thermal energy interchange during anaerobic digestion of waste organic solids at 36 C. Results over a period of 184 days indicated that the energy released during conversion of the waste substrate was utilized primarily for the production of CH(4) and that any excess released as heat during normal digestion was not sufficient to overcome heat losses from the system. The more exothermic response observed during retarded digestion was attributed to the accumulation of volatile acids and associated reduction in gas yields. After combustion of the CH(4) produced during digestion, between 2,370 and 3,950 kcal per lb of volatile solids converted per day was available for heating the process and maintaining optimal temperature conditions.     

Performances of a full-scale novel multiplate anaerobic reactor treating cheese whey effluent

A 450-m(3) multiplate anaerobic reactor (MPAR) has been started-up in April 1992 for treating wastewater (whey permeate and domestic wastewater) at the Nutrinor (Lactel) cheese factory in Chambord (Québec, Canada). The MPAR consists of four superimposed sections. The liquid flows upwards from one section to the next, while the gas is collected below each plate and evacuated through side-outlets. The wastewater is concurrently distributed at the bottom of the first, second, and third sections, as 50%, 33%, and 17% of the total influent stream, respectively. Granular anaerobic sludge at an initial concentration of 30 kg of volatile suspended solids (VSS) per cubic meter of reactor liquid volume was used to inoculate the reactor. Under normal operation of the factory, the chemical oxygen demand (COD) concentration of the influent ranged from 20 to 37 kg COD m(-3). The reactor organic loading rate (OLR) fluctuated between 9 and 14.7 kg COD m(-3) d(-1) for hydraulic retention times (HRT) maintained between 55 and 68 h. At the highest OLR, the MPAR showed an efficiency of 98% and 92% for soluble and total COD removal, respectively, and a methane production rate averaging around 4 m(3) m(-3) d(-1).Biomass-specific activities ranged between 7 and 51, 1.3 and 8.5, 5.3 and 12.2, 60 and 119, and 119 and 211 mmol g(-1) VSS d(-1) for glucose, propionate, acetate, formate, and hydrogen, respectively. Average equivalent-diameter of the granules was around 0.65 mm. The MPAR reactor generally showed a large capacity for solid retention with a biomass content between 32 and 37 kg VSS m(-3). (c) 1995 John Wiley & Sons, Inc.     

Fermentation and growth kinetic study of <Emphasis Type="Italic">Aeromonas caviae</Emphasis> under anaerobic conditions

Although Aeromonas caviae is pathogenic to a broad range of invertebrates including human, frequent in aquatic environments, and potentially vital for acidogenesis in anaerobic digestion, virtually no biokinetic information on its anaerobic growth is at hand. Therefore, this study focused on evaluating its anaerobic growth kinetics on glucose. To provide a set of relevant biokinetic coefficients for modeling, a combination of curve fitting and numerical modeling was used. Microcultivations were performed at eight different initial glucose concentrations of 0.1 to 2.5 g l⁻¹ to establish a function of specific growth rate versus substrate concentration. A batch anaerobic bioreactor was then operated to collect a data set for the numerical analysis. Kinetic coefficients were estimated from three different biomass growth profiles monitored by optical density, volatile suspended solids (VSS), or DNA measurement, and applied for simulating continuous operations at various hydraulic retention times (HRTs). Assuming the influent glucose concentration is 5,000 mg l⁻¹, the substrate utilization efficiency predicted to be 77.2% to 92.0% at 17 to 36 h HRTs. For the VSS-model-based simulation, the washout HRT was estimated to be 16.6 h, and similar for the other models. Overall, the anaerobic biokinetic coefficients of A. caviae grown on glucose were successfully estimated and found to follow a substrate inhibition model.     

Anaerobic digestion of processed municipal solid waste using a novel high solids reactor: Maximum solids levels and mixing requirements

Summary Novel, laboratory-scale, high solids reactors operated under mesophilic conditions were used to study the anaerobic fermentation of processed municipal solid waste (MSW) to methane. The anaerobic digestion consortium was introduced to high solids levels through gradual adaptation. The maximum sludge solids level for stable anaerobic fermentation performance was identified as approximately 36% wt/wt. Recovery of the anaerobic consortium, following dilution of inhibitory high solids levels, was swift. Reactor mixing requirements were also studied. No significant difference in fermentation performance was observed between agitator speeds of 1 and 25 rpm. Preliminary fermentation performance tests showed that solids loading rates as high as 9.5 g VS (volatile solids) feed/L sludge^.d, at 32% solids within the reactor, were possible. Under these conditions, operation was stable with an average pH of 7.8–8.0, total volatile fatty acid pools of <20 mM, and a biogas composition of 55%–60% methane.     

Anaerobic Digestion of Dairy Manure in a Hybrid Reactor with Biogas Recirculation

Summary A novel anaerobic hybrid reactor (AHR) configuration incorporating floating support media for biomass immobilization and biogas recirculation for enhanced mixing was used for anaerobic digestion of dairy manure. No pretreatment or solid liquid separation was applied. The reactor was operated at high influent volatile solids (VS) and organic loading rates (OLR) of up to 9.87% and 7.30 g VS/l day, respectively. After 149 days of continuous operation the results revealed that a high amount (38.1 g VSS) of biomass was able to attach itself to the support medium being used. The investigated AHR configuration achieved COD, BOD, TS, and VS removal efficiencies of 48–63, 64–78, 55–65, and 59–68%, respectively, at a hydraulic retention time (HRT) of 15 days. The corresponding average methane production value obtained in this study was 0.191 l/g VS added.     

Bacterial community analysis in upflow multilayer anaerobic reactor treating high‐solids organic wastes

A novel anaerobic digestion configuration, the upflow multi‐layer anaerobic reactor (UMAR), was developed to treat high‐solids organic wastes. The UMAR was hypothesized to form multi‐layer along depth due to the upflow plug flow; use of a recirculation system and a rotating distributor and baffles aimed to assist treating high‐solids influent. The chemical oxygen demand (COD) removal efficiency and methane (CH₄) production rate were 89% and 2.10 L CH₄/L/d, respectively, at the peak influent COD concentration (110.4 g/L) and organic loading rate (7.5 g COD/L/d). The 454 pyrosequencing results clearly indicated heterogeneous distribution of bacterial communities at different vertical locations (upper, middle, and bottom) of the UMAR. Firmicutes was the dominant (>70%) phylum at the middle and bottom parts, while Deltaproteobacteria and Chloroflexi were only found in the upper part. Potential functions of the bacteria were discussed to speculate on their roles in the anaerobic performance of the UMAR system. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1226–1234, 2017     

Lab-scale study of an anaerobic membrane bioreactor (AnMBR) for dilute municipal wastewater treatment

Anaerobic bioreactors supplemented with membrane technology have become quite popular, owing to their favorable energy recovery characteristics. In this study, a lab-scale anaerobic Membrane Bioreactor (AnMBR) was assessed in experimental treatments of pre-settled dilute municipal wastewater obtained from a full-scaled wastewater treatment plant. The MBR system was operated in continuous flow mode for 440 days. To evaluate the performance of the AnMBR under various loading rates, the hydraulic retention time (HRT) was reduced in a stepwise manner (from 2 to 0.5 days). Afterward, the mixed liquor suspended solids (MLSS) were reduced from 7,000 to 3,000 mg/L in increments of 1,000 mg/L, resulting in a decrease in solids retention time (SRT) at a constant HRT of 1.0 day. The soluble chemical oxygen demand (SCOD) concentration in the feed varied between 38 and 131 mg/L, whereas the average permeate SCOD ranged between 18 and 37 mg/L, reflecting excellent effluent quality. The AnMBR performance in terms of COD removal proved stable, despite variations in influent characteristics and HRT and SRT changes. The concentration of extracellular polymeric substance (EPS) was reduced with decreases in HRT from 42 to 22 mg VS/mg of MLSS, thereby indicating that the increased biomass concentration biodegraded the EPS at lower HRTs. AnMBR is, therefore, demonstrably a feasible option for the treatment of dilute wastewater with separate stage nitrogen and phosphorus removal processes.     

Two-phase partitioning bioreactors in environmental biotechnology

Operation of microbial electrolysis cells (MECs) without an ion exchange membrane could help to lower the construction costs while lowering the ohmic cell resistance and improving MEC conversion rates by minimizing the pH gradient between anode and cathode. In this research, we demonstrate that membraneless MECs with plain graphite can be operated for methane production without pH adjustment and that the ohmic cell resistance could be lowered with approximately 50% by removing the cation exchange membrane. As a result, the current production increased from 66 ± 2 to 156 ± 1 A m⁻³ MEC by removing the membrane with an applied voltage of −0.8 V. Methane was the main energetic product despite continuous operation under carbonate-limited and slightly acidified conditions (pH 6.1–6.2). Our results suggest that continuous production of hydrogen in membraneless MECs will be challenging since methane production might not be avoided easily. The electrical energy invested was not always completely recovered under the form of an energy-rich biogas; however, our results indicate that membraneless MECs might be a viable polishing step for the treatment of the effluent of anaerobic digesters as methane was produced under low organic loading conditions and at room temperature. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Application of redox mediators to accelerate the transformation of reactive azo dyes in anaerobic bioreactors.

Azo dyes are nonspecifically reduced under anaerobic conditions but the slow rates at which reactive azo dyes are converted presents a serious problem for the application of anaerobic technology as a first stage in the complete biodegradation of these compounds. As quinones have been found to catalyze reductive transfers by acting as redox mediators, the application of anthraquinone-2,6-disulfonic acid (AQDS) during continuous anaerobic treatment of the reactive azo dye, Reactive Red 2 (RR2), was evaluated. A mixture of volatile fatty acids was used as the electron-donating primary substrate. Batch experiments demonstrated that AQDS could increase the first-order rate constant of RR2 reductive cleavage by one order of magnitude. In the continuous experiment, treatment of RR2 containing synthetic wastewater in a lab-scale upflow anaerobic sludge blanket (UASB) reactor yielded low dye removal efficiencies (<30%). Consequently, severe toxicity problems occurred, eventually resulting in almost complete inhibition of the methanogenic activity. Addition of catalytic concentrations of AQDS (19 microM) to the reactor influent caused an immediate increase in the dye removal efficiency and recovery of biological activity. Ultimately, RR2 removal efficiency stabilized at 88%, and higher AQDS loads resulted in higher RR2 removal efficiencies (up to 98% at 155 microM AQDS). Examination of the RR2 decolorizing properties of dye-adapted reactor sludge and of nonadapted reactor seed sludge revealed that RR2 decolorization was principally a biologically driven transfer of reducing equivalents from endogenous and added substrates to the dye. Hydrogen, added in bulk, was clearly the preferred electron donor. Bacteria that couple dye decolorization to hydrogen oxidation were naturally present in seed sludge. However, enrichment was required for the utilization of electrons from volatile fatty acids for dye reduction. The stimulatory effect of AQDS on RR2 decolorization by AQDS-unadapted sludge was mainly due to assisting the electron transfer from endogenous substrates in the sludge to the dye. The stimulatory effect of AQDS on RR2 decolorization by sludge from the AQDS-exposed reactor was, in addition, strongly associated with the transfer of electrons from hydrogen and acetate to the dye, probably due to enrichment of specialized AQDS-reducing bacteria.     

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO₂, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic‐ and thermophilic anaerobic cultures were enriched to convert CO₂ to CH₄ by addition of H₂. Enrichment at thermophilic temperature (55°C) resulted in CO₂ and H₂ bioconversion rate of 320 mL CH₄/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic‐ and thermophilic‐enriched cultures, as revealed by PCR–DGGE. Nonetheless, they all belonged to the order Methanobacteriales, which can mediate hydrogenotrophic methanogenesis. Biogas upgrading was then tested in a thermophilic anaerobic reactor under various operation conditions. By continuous addition of hydrogen in the biogas reactor, high degree of biogas upgrading was achieved. The produced biogas had a CH₄ content, around 95% at steady‐state, at gas (mixture of biogas and hydrogen) injection rate of 6 L/(L day). The increase of gas injection rate to 12 L/(L day) resulted in the decrease of CH₄ content to around 90%. Further study showed that by decreasing the gas–liquid mass transfer by increasing the stirring speed of the mixture the CH₄ content was increased to around 95%. Finally, the CH₄ content around 90% was achieved in this study with the gas injection rate as high as 24 L/(L day). Biotechnol. Bioeng. 2012; 109: 2729–2736. © 2012 Wiley Periodicals, Inc.     

Influence of substrate concentration on the stability and yield of continuous biohydrogen production

The effect of substrate concentration (sucrose) on the stability and yield of a continuous fermentative process producing hydrogen was studied. High substrate concentrations are attractive from an energy standpoint as they would minimise the energy required for heating. The reactor was a CSTR; temperature was maintained at 35 degrees C; pH was controlled between 5.2 and 5.3, and the hydraulic retention time (HRT) was 12 h. Online measurements were taken for ORP, pH, temperature, %CO2, gas output and %H2, and data logged using a MatLAB data acquisition toolbox. Steady-state operation was obtained at 10, 20 and 40 g/L of sucrose in the influent, but a subsequent step change to 50 g/L was unsustainable. The hydrogen content ranged between 50% and 60%. The yield of hydrogen decreased as the substrate concentration increased from 1.7 +/- 0.2 mol/mol hexose added at 10 g/L, to 0.8 +/- 0.1 mol/mol at 50 g/L. Sparging with nitrogen improved the hydrogen yield by at least 35% at 40 g/L and at least 33% at 50 g/L sucrose. Sparging also enabled steady-state operation at 50 g/L sucrose. Addition of an extra 4 g/L of n-butyric acid to the reactor operating at 40 g/L sucrose increased the butyrate concentration from 9,830 to 18,900 mg/L, immediately stopping gas production and initiating the production of propionate, whilst the addition of 2 g/L taking the butyrate concentration to 12,200 mg/L did not do so. It was shown that operation at 50 g/L sucrose in a CSTR in butyrate fermentation is possible.     

Evaluation of startup and operation of four anaerobic processes treating a synthetic meat waste

Two continuous stirred tanks reactors (CSTR) and four anaerobic fluidized bed reactors (AFBR) were used to study the treatment of a synthetic meat waste during single-and two-stage anaerobic treatment. Four configurations were investigated; a single-stage CSTR and AFBR and the two-stage systems CSTR-AFBR and AFBR-AFBR. Startup of the anaerobic reactors was achieved within 50 days by use of a regime that included stepped increases in influent COD, methanol substitution of the substrate, and addition of essential trace metals such as cobalt and nickel. Two-stage reactors removed up to 85% of influent COD concentrations of 5000 mg/L, whereas the single-stage AFBR and CSTR removed 76 and 9%, respectively. The proportion of methane in the effluent gases increased as the influent COD concentration was increased. Volumetric production of methane was greatest for the first stage of the AFBR-AFBR system. Solids retention times calculated for the AFBRs ranged from 7 to 12 days, sufficient to support methanogenesis. The AFBRs and two-stage systems were more resistant to an influent pH shock from the operating value of pH 6.8 down to pH 3 than the CSTRs and single-stage reactors. It was concluded that high-rate anaerobic treatment systems were applicable to meat industry wastewaters and that two-stage digestion produced a better quality effluent.     

Enhancement of hydrogen production in a single chamber microbial electrolysis cell through anode arrangement optimization

Reducing the inner resistances is crucial for the enhancement of hydrogen generation in microbial electrolysis cells (MECs). This study demonstrates that the optimization of the anode arrangement is an effective strategy to reduce the system resistances. By changing the normal MEC configuration into a stacking mode, namely separately placing the contacted anodes from one side to both sides of cathode in parallel, the solution, biofilm and polarization resistances of MECs were greatly reduced, which was also confirmed with electrochemical impedance spectroscopy analysis. After the anode arrangement optimization, the current and hydrogen production rate (HPR) of MEC could be enhanced by 72% and 118%, reaching 621.3 ± 20.6 A/m³ and 5.56 m³/m³ d respectively, under 0.8 V applied voltage. A maximum current density of 1355 A/m³ with a HPR of 10.88 m³/m³ d can be achieved with 1.5 V applied voltage.     

Analysis of ammonia loss mechanisms in microbial fuel cells treating animal wastewater

Ammonia losses during swine wastewater treatment were examined using single- and two-chambered microbial fuel cells (MFCs). Ammonia removal was 60% over 5 days for a single-chamber MFC with the cathode exposed to air (air-cathode), versus 69% over 13 days from the anode chamber in a two-chamber MFC with a ferricyanide catholyte. In both types of systems, ammonia losses were accelerated with electricity generation. For the air-cathode system, our results suggest that nitrogen losses during electricity generation were increased due to ammonia volatilization with conversion of ammonium ion to the more volatile ammonia species as a result of an elevated pH near the cathode (where protons are consumed). This loss mechanism was supported by abiotic tests (applied voltage of 1.1 V). In a two-chamber MFC, nitrogen losses were primarily due to ammonium ion diffusion through the membrane connecting the anode and cathode chambers. This loss was higher with electricity generation as the rate of ammonium transport was increased by charge transfer across the membrane. Ammonia was not found to be used as a substrate for electricity generation, as intermittent ammonia injections did not produce power. The ammonia-oxidizing bacterium Nitrosomonas europaea was found on the cathode electrode of the single-chamber system, supporting evidence of biological nitrification, but anaerobic ammonia-oxidizing bacteria were not detected by molecular analyses. It is concluded that ammonia losses from the anode chamber were driven primarily by physical-chemical factors that are increased with electricity generation, although some losses may occur through biological nitrification and denitrification.