Bioelectrochemical extraction of ammonium from low-strength wastewater with concomitant generation of high-purity hydrogen.

Bioelectrochemical systems (BES) are emerging environmental biotechnology for recovering ammonia from waste streams. It has been tested extensively for treating ammonium-rich wastewater. This study examined the suitability of BES to facilitate carbon removal and ammonium extraction from a low ammonium liquor (3.7 mM) that mimics municipal wastewater, and concomitant production of high-purity hydrogen gas, which could potentially be harnessed as a fuel or internally recycled for ammonia stripping. Results showed that a two-chamber cation exchange membrane-equipped BES enabled a high hydrogen yield (22.8 m3 H2 m-3 d-1; > 98% cathodic efficiency) and chemical oxygen demand (COD) removal (80%; 2.43 kg COD m-3 d-1 at a hydraulic retention time of 4.4 h). However, for the treatment of wastewater, the system demanded high energy (2.3 kWh kg COD-1; 152 kWh kg-1 N removed) and base for pH adjustment. The technology may be more suitable for recovering ammonium from wastewaters with molar ammonium to BOD ratio closer to the desired stoichiometric ratio of four, and for waste streams containing sufficient alkalinity or pH-buffering capacity, eliminating the need for dosing cation-bearing alkali.

Sustained and enhanced anaerobic removal of COD and nitrogen in a zeolite amended glycogen accumulating organism dominated biofilm process.

Activated sludge, the most widely used biological wastewater treatment process is known to be expensive to operate, largely due to energy expense for oxygen transfer into the bulk wastewater solution. The alternative of using passive aeration facilitates oxygen supply directly from the air resulting in aeration energy savings. The current study demonstrated sustained and improved removal of chemical oxygen demand (COD) and nitrogen in a zeolite modified glycogen accumulating organisms (GAOs) dominated biofilm reactor, which achieved anaerobic removal of COD and ammonium by the activity of GAOs and zeolite, respectively. Draining of the batch-operated reactor enabled the biofilm to directly uptake oxygen from air (passive aeration) to carry out simultaneous nitrification and denitrification due to the activity of GAO (Candidatus competibacter) and nitrifying bacteria (Nitrosomonas and Nitrospira). Under stable long-term (4-months) operation, the process achieved COD and nitrogen removal at rates of 1354 and 79.1 g m-3 d-1, respectively. The biofilm process demonstrated >90% nitrogen removal efficiency in multi-cycle (4/8 cycles) strategy with a short treatment time of 8 h. Due to the passive aeration scheme, the energy consumption of the proposed wastewater treatment process is calculated to be about 13-times less than that of traditional activated sludge process. Therefore, the Passive Aeration Simultaneous Nitrification and Denitrification (PASND) biofilm process is a promising low-energy treatment step for efficient removal of COD and nitrogen from wastewater.

Novel microbial-electrochemical filter with a computer-feedback pH control strategy for upgrading biogas into biomethane.

A novel microbial-electrochemical filter was designed and operated based on a combined microbial electrolysis cell and bio-trickling filter principles with the aim to maximize gas-liquid mass-transfer efficiency and minimize costs associated with bubbling biogas through liquid-filled reactor. CO2/biogas feed to the MEF was done via a computer-feedback pH control strategy, linking CO2 feed directly to the OH- production. As a result current efficiency was constant at around 100% throughout the period of experiments. CO2 from biogas was almost completely removed at cathodic pH setpoint of 8.5. Maximum CO2 removal rate was 14.6 L/L/day (equivalent to 29.2 L biogas/L/day). Net energy consumption was around 1.28 kWh/Nm3CO2 or 0.64 kWh/m3 biogas (maximum 49% energy efficiency). An ability to maintain a constant pH means elevated pH from increasing applied potential (current) is no longer an issue. The process can potentially be up-scaled and operated at a much higher current and therefore CO2 removal rate.

Energy efficient COD and N-removal from high-strength wastewater by a passively aerated GAO dominated biofilm.

Conventional aerobic treatment of high-strength wastewater is not economical due to excessively high energy requirement for compressed air supply. The use of passive aeration avoids the use of compressed air and enables energy efficient oxygen supply directly from the air. This study evaluates a passively aerated simultaneous nitrification and denitrification performing biofilm to treat concentrated wastewater. The biofilm reactor was operated > 5-months under alternating anaerobic/aerobic conditions. For 4-times concentrated wastewater, > 80% COD (2307 mg L-1 h-1) and > 60% N (60 mg L-1 h-1) was removed at a hydraulic retention time (HRT) of 7 h. A double application in the same reactor enabled > 95% COD and 85% N-removal, at an overall HRT of 14 h which is substantially shorter than what traditional activated sludge-based systems would require for the treatment of such concentrated feeds. Microbial community analysis showed Candidatus competibacter (27%) and nitrifying bacteria (Nitrosomonas, and Nitrospira) as key microbes involved in COD and N-removal, respectively.

Anaerobic acidification of sugar-containing wastewater for biotechnological production of organic acids and ethanol.

Anaerobic acidification of sugars can produce some useful end-products such as alcohol, volatile fatty acids (e.g. acetate, propionate, and butyrate) and lactic acid. The production of end-products is highly dependent on factors including pH, temperature, hydraulic retention time and the types of sugar being fermented. Results of this current study indicate that the pH and hydraulic retention time played significant roles in determining the end products from the anaerobic acidification of maltose and glucose. Under uncontrolled pH, the anaerobic acidification of maltose ceased when pH in the reactor dropped below 5 while anaerobic acidification of glucose continued and produced ethanol as the main end-product. Under controlled pH, lactic acid was found to be the dominant end-product produced from both maltose and glucose at pH 5. Acetate was the main end-product from both maltose and glucose fermented at neutral pH (6 and 7). Short hydraulic retention time (HRT) of 2 days could induce the production of ethanol from the anaerobic acidification of glucose. However, the anaerobic acidification of maltose could stop when short HRT of 2 days was applied in the reactor. This finding is significant for industrial fermentation and waste management systems, and selective production of different types of organic acids could be achieved by managing pH and HRT in the reactor.

Direct oxygen uptake from air by novel glycogen accumulating organism dominated biofilm minimizes excess sludge production.

The cost associated with treatment and disposal of excess sludge produced is one of the greatest operational expenses in wastewater treatment plants. In this study, we quantify and explain greatly reduced excess sludge production in the novel glycogen accumulating organism (GAO) dominated drained biofilm system previously shown to be capable of extremely energy efficient removal of organic carbon (biological oxygen demand or BOD) from wastewater. The average excess sludge production rate was 0.05 g VSS g-1 BOD (acetate) removed, which is about 9-times lower than that of comparative studies using the same acetate based synthetic wastewater. The substantially lower sludge yield was attributed to a number of features such as the high oxygen consumption facilitated by direct oxygen uptake from air, high biomass content (21.41 g VSS L-1 of reactor), the predominance of the GAO (Candidatus competibacter) with a low growth yield and the overwhelming presence of the predatory protozoa (Tetramitus) in the biofilm. Overall, the combination of low-energy requirement for air supply (no compressed air supply) and the low excess sludge production rate, could make this novel "GAO drained biofilm" process one of the most economical ways of biological organic carbon removal from wastewater.

Ethanol and lactic acid production from sugar and starch wastes by anaerobic acidification.

Anaerobic conversion of carbohydrates can generate various end-products. Besides physical parameters such as pH and temperature, the types of carbohydrate being fermented influences the fermentation pattern. Under uncontrolled pH, microbial mixed cultures from activated sludge and anaerobic digester sludge anaerobically produced ethanol from glucose while producing lactic acid from starch conversion. This trend was not only observed in batch trials. Also, continuous chemostat operation of anaerobic digester sludge resulted in the reproducible predominance of ethanol fermentation from glucose solution and lactic acid production from starch. Different feeding regimes and substrate availability (shock load versus continuous feeding) in glucose fermentation under non-controlled pH did not affect the ethanol production as the major end product. Shifts in feed composition from glucose to starch and vice versa result in an immediate change of fermentation end products formation.

Proof of concept of wastewater treatment via passive aeration SND using a novel zeolite amended biofilm reactor.

The current paper describes a novel passive aeration simultaneous nitrification and denitrification (PASND) zeolite amended biofilm reactor that removes organic carbon and nitrogen from wastewater with low-energy consumption. Next to the ammonium oxidizing bacteria (AOB), this reactor contained naturally enriched glycogen accumulating organisms (GAOs) and zeolite powder to initially adsorb BOD (acetate) and ammonium (NH4 +-N) from synthetic wastewater under anaerobic conditions. Draining of the treated wastewater exposed the biofilm directly to air enabling low-energy oxygen supply by passive aeration. This allowed the adsorbed ammonium to be oxidized by the AOB and the produced nitrite and nitrate to be reduced simultaneously by the GAOs using the adsorbed BOD (stored as PHAs) as carbon source. Overall, with an operation mode of 1 h anaerobic and 4 h aerobic phase, the nutrient removal efficiency after single treatment was about 94.3% for BOD and 72.2% for nitrogen (NH4 +-N). As high-energy aeration of the bulk solution for oxygen supply is completely avoided, the energy requirement of the proposed PASND biofilm reactor can be theoretically cut down to more than 50% compared to the traditional activated sludge process.

Concurrent Lactic and Volatile Fatty Acid Analysis of Microbial Fermentation Samples by Gas Chromatography with Heat Pre-treatment.

Organic acid analysis of fermentation samples can be readily achieved by gas chromatography (GC), which detects volatile organic acids. However, lactic acid, a key fermentation acid is non-volatile and can hence not be quantified by regular GC analysis. However the addition of periodic acid to organic acid samples has been shown to enable lactic acid analysis by GC, as periodic acid oxidizes lactic acid to the volatile acetaldehyde. Direct GC injection of lactic acid standards and periodic acid generated inconsistent and irreproducible peaks, possibly due to incomplete lactic acid oxidation to acetaldehyde. The described method is developed to improve lactic acid analysis by GC by using a heat treated derivatization pre-treatment, such that it becomes independent of the retention time and temperature selection of the GC injector. Samples containing lactic acid were amended by periodic acid and heated in a sealed test tube at 100°C for at least 45 min before injecting it to the GC. Reproducible and consistent peaks of acetaldehyde were obtained. Simultaneous determination of lactic acid, acetone, ethanol, butanol, volatile fatty acids could also be accomplished by applying this GC method, enabling precise and convenient organic acid analysis of biological samples such as anaerobic digestion and fermentation processes.

Bioelectrochemical enhancement of anaerobic digestion: Comparing single- and two-chamber reactor configurations at thermophilic conditions.

Bioelectrochemical system (BES) can act as an auxiliary technology for improving organic waste treatment and biogas production in anaerobic digestion (AD). For the first time this study directly compared the performance of a single- and a cation-exchange membrane-equipped two-chamber BES-AD systems at thermophilic conditions. The results indicated that an active glucose-fed thermophilic anaerobic sludge could readily (<3days) increase biogas production in both reactor configurations by inserting a carbon electrode poised at -0.8V (vs. Ag/AgCl). However, after a 3-week operation, the biogas production rates from the single- and two-chamber BES reactor decreased due to volatile fatty acids accumulation. Only the two-chamber configuration could enable methane enrichment (98% CH4v/v) in biogas. Overall, this study suggests that integrating bioelectrodes in-situ could not sustainably improve biogas production in a thermophilic AD reactor, and future studies should be directed towards the use of bioelectrodes for improving biogas quality.

Rapid adaptation of activated sludge bacteria into a glycogen accumulating biofilm enabling anaerobic BOD uptake.

Glycogen accumulating organisms (GAO) are known to allow anaerobic uptake of biological oxygen demand (BOD) in activated sludge wastewater treatment systems. In this study, we report a rapid transition of suspended activated sludge biomass to a GAO dominated biofilm by selective enrichment using sequences of anaerobic loading followed by aerobic exposure of the biofilm to air. The study showed that within eight weeks, a fully operational, GAO dominated biofilm had developed, enabling complete anaerobic BOD uptake at a rate of 256mg/L/h. The oxygen uptake by the biofilm directly from the atmosphere had been calculated to provide significant energy savings. This study suggests that wastewater treatment plant operators can convert activated sludge systems readily into a "passive aeration" biofilm that avoids costly oxygen transfer to bulk wastewater solution. The described energy efficient BOD removal system provides an opportunity to be coupled with novel nitrogen removal processes such as anammox.

New method for characterizing electron mediators in microbial systems using a thin-layer twin-working electrode cell.

Microbial biofilms are significant ecosystems where the existence of redox gradients drive electron transfer often via soluble electron mediators. This study describes the use of two interfacing working electrodes (WEs) to simulate redox gradients within close proximity (250µm) for the detection and quantification of electron mediators. By using a common counter and reference electrode, the potentials of the two WEs were independently controlled to maintain a suitable "voltage window", which enabled simultaneous oxidation and reduction of electron mediators as evidenced by the concurrent anodic and cathodic currents, respectively. To validate the method, the electrochemical properties of different mediators (hexacyanoferrate, HCF, riboflavin, RF) were characterized by stepwise shifting the "voltage window" (ranging between 25 and 200mV) within a range of potentials after steady equilibrium current of both WEs was established. The resulting differences in electrical currents between the two WEs were recorded across a defined potential spectrum (between -1V and +0.5V vs. Ag/AgCl). Results indicated that the technique enabled identification (by the distinct peak locations at the potential scale) and quantification (by the peak of current) of the mediators for individual species as well as in an aqueous mixture. It enabled a precise determination of mid-potentials of the externally added mediators (HCF, RF) and mediators produced by pyocyanin-producing Pseudomonas aeruginosa (WACC 91) culture. The twin working electrode described is particularly suitable for studying mediator-dependent microbial electron transfer processes or simulating redox gradients as they exist in microbial biofilms.

Automatic online buffer capacity (alkalinity) measurement of wastewater using an electrochemical cell.

The use of an automatic online electrochemical cell (EC) for measuring the buffer capacity of wastewater is presented. pH titration curves of different solutions (NaHCO3, Na2HPO4, real municipal wastewater, and anaerobic digester liquid) were obtained by conventional chemical titration and compared to the online EC measurements. The results show that the pH titration curves from the EC were comparable to that of the conventional chemical titration. The results show a linear relationship between the response of the online EC detection system and the titrimetric partial alkalinity and total alkalinity of all tested samples. This suggests that an EC can be used as a simple online titration device for monitoring the buffer capacity of different industrial processes including wastewater treatment and anaerobic digestion processes.

Novel process of bio-chemical ammonia removal from air streams using a water reflux system and zeolite as filter media.

A novel biofilter that removes ammonia from air streams and converts it to nitrogen gas has been developed and operated continuously for 300 days. The ammonia from the incoming up-flow air stream is first absorbed into water and the carrier material, zeolite. A continuous gravity reflux of condensed water from the exit of the biofilter provides moisture for nitrifying bacteria to develop and convert dissolved ammonia (ammonium) to nitrite/nitrate. The down-flow of the condensed water reflux washes down nitrite/nitrate preventing ammonium and nitrite/nitrate accumulation at the top region of the biofilter. The evaporation caused by the inflow air leads to the accumulation of nitrite to extremely high concentrations in the bottom of the biofilter. The high nitrite concentrations favour the spontaneous chemical oxidation of ammonium by nitrite to nitrogen (N2). Tests showed that this chemical reaction was catalysed by the zeolite filter medium and allowed it to take place at room temperature. This study shows that ammonia can be removed from air streams and converted to N2 in a fully aerated single step biofilter. The process also overcomes the problem of microorganism-inhibition and resulted in zero leachate production.

Simultaneous phosphorus uptake and denitrification by EBPR-r biofilm under aerobic conditions: effect of dissolved oxygen.

A biofilm process, termed enhanced biological phosphorus removal and recovery (EBPR-r), was recently developed as a post-denitrification approach to facilitate phosphorus (P) recovery from wastewater. Although simultaneous P uptake and denitrification was achieved despite substantial intrusion of dissolved oxygen (DO >6 mg/L), to what extent DO affects the process was unclear. Hence, in this study a series of batch experiments was conducted to assess the activity of the biofilm under various DO concentrations. The biofilm was first allowed to store acetate (as internal storage) under anaerobic conditions, and was then subjected to various conditions for P uptake (DO: 0-8 mg/L; nitrate: 10 mg-N/L; phosphate: 8 mg-P/L). The results suggest that even at a saturating DO concentration (8 mg/L), the biofilm could take up P and denitrify efficiently (0.70 mmol e(-)/g total solids*h). However, such aerobic denitrification activity was reduced when the biofilm structure was physically disturbed, suggesting that this phenomenon was a consequence of the presence of oxygen gradient across the biofilm. We conclude that when a biofilm system is used, EBPR-r can be effectively operated as a post-denitrification process, even when oxygen intrusion occurs.

Organic carbon removal from wastewater by a PHA storing biofilm using direct atmospheric air contact as oxygen supply.

The principal reason for the high energy costs for biological wastewater treatment is the poor transfer efficiency of oxygen to the bulk water phase. The current paper describes a biofilm reactor in which oxygen transfer to the bulk solution is avoided by alternating anaerobic submersed (2h) and drained (1h) operation of the biofilm. During the submersed phase the biofilm enriched for glycogen accumulating organism (GAO) stored the organic carbon (acetate) as poly-hydroxy-alkanoate (PHA). After draining the reactor, this carbon stored as PHA was biologically oxidised, using oxygen directly from the atmosphere. The 12Cmmol/L (384mg/L BOD) of acetate was completely removed during long term automated operation of the reactor for 9months with a cycle length of 3.3h. As the process specifically removes dissolved organic carbon but not N or P it could possibly be coupled with novel processes such as Anammox or nutrient recovery.

Microbial fuel cell biosensor for rapid assessment of assimilable organic carbon under marine conditions.

The development of an assimilable organic carbon (AOC) detecting marine microbial fuel cell (MFC) biosensor inoculated with microorganisms from marine sediment was successful within 36 days. This established marine MFC was tested as an AOC biosensor and reproducible microbiologically produced electrical signals in response to defined acetate concentration were achieved. The dependency of the biosensor sensitivity on the potential of the electron-accepting electrode (anode) was investigated. A linear correlation (R(2) > 0.98) between electrochemical signals (change in anodic potential and peak current) and acetate concentration ranging from 0 to 150 µM (0-3600 µg/L of AOC) was achieved. However, the present biosensor indicated a different-linear relation at somewhat elevated acetate concentration ranging from 150 to 450 µM (3600-10,800 µg/L of AOC). This high concentration of acetate addition could be measured by coulombic measurement (cumulative charges) with a linear correlation. For the acetate concentration detected in this study, the sensor recovery time could be controlled within 100 min.

In-line deoxygenation for organic carbon detections in seawater using a marine microbial fuel cell-biosensor.

Assimilable organic carbon (AOC) is a key predictor for membrane biofouling in seawater desalination reverse osmosis (SWRO). Microbial fuel cells have been considered as biosensors for the detection of biodegradable organics. However, the presence of dissolved oxygen (DO) is known to completely suppress the signal production (i.e., current) of a typical MFC. This study describes AOC detection in normal oxygenated seawater by coupling an electrochemical cell for DO removal with a MFC-biosensor for AOC detection. The electrochemical deoxygenation for oxygen removal caused no interference in the AOC detection. A linear relationship (R(2)=0.991) between the AOC concentration and current production from the MFC biosensor was achieved. The coupling of an electrochemical cell with a MFC-biosensor can be effectively used as an online, rapid and inexpensive measure of AOC concentrations and hence as an indicator for biofouling potential of seawater.

Hexacyanoferrate-adapted biofilm enables the development of a microbial fuel cell biosensor to detect trace levels of assimilable organic carbon (AOC) in oxygenated seawater.

A marine microbial fuel cell (MFC) type biosensor was developed for the detection of assimilable organic carbon (AOC) in ocean water for the purpose of online water quality monitoring for seawater desalination plants prone to biofouling of reverse osmosis (RO) membranes. The anodophilic biofilm that developed on the graphite tissue anode could detect acetate as the model AOC to concentrations as low as 5 µM (120 µg/L of AOC), which is sufficiently sensitive as an online biofouling risk sensor. Although the sensor was operated at a higher (+200 ± 10 mV) than the usual (-300 mV) anodic potential, the presence of oxygen completely suppressed the electrical signal. In order to overcome this outcompeting effect of oxygen over the anode as electron acceptor by the bacteria, hexacyanoferrate (HCF(III)) was found to enable the development of an adapted biofilm that transferred electrons to HCF(III) rather than oxygen. As the resultant of the reduced HCF(II) could readily transfer electrons to the anode while being re-oxidised to HCF(III), the marine MFC biosensor developed could be demonstrated to work in the presence of oxygen unlike traditional MFC. The possibility of operating the marine MFC in batch or continuous (in-line) mode has been explored by using coulombic or potentiometric interpretation of the signal.

Selective enrichment and production of highly urease active bacteria by non-sterile (open) chemostat culture.

In general, bioprocesses can be subdivided into naturally occurring processes, not requiring sterility (e.g., beer brewing, wine making, lactic acid fermentation, or biogas digestion) and other processes (e.g., the production of enzymes and antibiotics) that typically require a high level of sterility to avoid contaminant microbes overgrowing the production strain. The current paper describes the sustainable, non-sterile production of an industrial enzyme using activated sludge as inoculum. By using selective conditions (high pH, high ammonia concentration, and presence of urea) for the target bacterium, highly active ureolytic bacteria, physiologically resembling Sporosarcina pasteurii were reproducibly enriched and then continuously produced via chemostat operation of the bioreactor. When using a pH of 10 and about 0.2 M urea in a yeast extract-based medium, ureolytic bacteria developed under aerobic chemostat operation at hydraulic retention times of about 10 h with urease levels of about 60 µmol min⁻¹ ml⁻¹ culture. For cost minimization at an industrial scale the costly protein-rich yeast extract medium could be replaced by commercial milk powder or by lysed activated sludge. Glutamate, molasses, or glucose-based media did not result in the enrichment of ureolytic bacteria by the chemostat. The concentration of intracellular urease was sufficiently high such that the produced raw effluent from the reactor could be used directly for biocementation in the field.