WO2019079908A1

WO2019079908A1 - Bioelectrochemical method and apparatus for energy reclamation from nitrogen compounds

Info

Publication number: WO2019079908A1
Application number: PCT/CA2018/051366
Authority: WO
Inventors: Michael Siegert
Original assignee: Michael Siegert
Priority date: 2017-10-29
Filing date: 2018-10-29
Publication date: 2019-05-02
Also published as: US11691901B2; EP3700868A4; CA3080819A1; US20200339453A1; EP3700868A1

Abstract

Methods are described for treating aqueous solutions, including wastewater, to remove nitrogen-containing compounds using electrochemical processes. The method may be conducted electrolytically under an applied voltage or using endogenous current in a fuel cell arrangement. In some embodiments, energy is reclaimed in the form of hydrogen, methane, and other hydrocarbons or organic molecules. Microorganisms may be used as the catalyst for oxidation of the nitrogen-containing compound and/or reduction of hydrogen ions, carbon dioxide, or bicarbonate. Anaerobic or low-oxygen conditions may be used in the zone.

Description

BIOELECTROCHEMICAL METHOD AND APPARATUS FOR ENERGY

RECLAMATION FROM NITROGEN COMPOUNDS

[0001] This application claims priority based on U .S. Application No.

62/578,456 entitled "Processes for Energy Reclamation from Nitrogen Removal From Liquids and for Electrical Current Production" filed on October 29, 2017, which is herein incorporated by reference in its entirety.

TECHNICAL FIELD

[002] The present disclosure relates to the field of nitrogen removal from solutions. More specifically, the present disclosure relates to the removal of ammonium or amines of wastewater.

BACKGROUND

[003] Wastewater and other solutions often contain nitrogen- containing compounds that must be disposed of. For wastewater, ammonium removal is important for proper discharge in order to minimize environmental damage. Other disposal needs also arise in situations where nitrogen- containing compounds are dissolved in solution, but no longer wanted.

[004] Conventional approaches for removing nitrogen from aqueous solutions rely on oxygen saturation in order to convert nitrogen-containing compounds, such as ammonium and amines, into nitrate (nitrification) which subsequently converted into gaseous nitrogen ( N2, denitrification). However, oxygen does not dissolve well in solution, making nitrogen removal an energy intensive process. Moreover, oxygen is a strong oxidant, which consumes much of the available chemical energy as heat, resulting in a process that is energy inefficient.

[005] A particular challenge of oxidizing nitrogen-containing

compounds is the strong bond energy that all nitrogen atoms have. In biological systems, this is overcome by using strong oxidants such as O2, NO3^", NO2^". Alternative approaches for nitrogen removal of aqueous solutions still require the presence of oxygen, either directly or indirectly.

[006] For example, in the "annamox" process, a portion of the ammonium ions are first oxidized to nitrite using molecular oxygen.

Microorganisms then react the nitrite ions with the balance of the ammonium ions in solution, to produce nitrogen gas (N2) . However, this process is slow, requires high solution volumes, operates under a narrow range of temperature and pH, requires excess nitrate or nitrite that must still be disposed of, and does not recapture energy from ammonium.

[007] Another approach for nitrogen removal is microbial electrolysis, in which one or more electrodes are covered in a biofilm or are placed in a suspension of microorganisms. Examples of this approach are described by Kuntke et al. in U .S. Patent No. 9,725,812 and Kuroda et al. in European Patent No. 0573226. In such approaches, the microorganisms that remove

contaminants from the water are offered a small energetic incentive in the form of an electrochemical potential, which they can use for their metabolism. In exchange, the microorganisms catalyse the oxidation of ammonium, nitrate, or nitrite into N2. Nevertheless, such approaches also rely on aerated conditions in order to achieve complete nitrogen removal, which substantially reduces the energy available for recapture of useful compounds from the treated solution, such as hydrogen gas, at the cathode. Moreover, the complexity of such systems, which typically require three or more electrochemically active electrodes, makes them expensive to manufacture and operate. Finally, such systems have not been successful in removing nitrogen at lower concentrations, such as ammonium concentrations below 0.5 g/L.

[008] Various compounds of interest may also be recovered from aqueous solutions during electrolysis. For example, hydrogen gas can be produced at the cathode by reducing FT ions in water, whereas methane can be produced at the cathode by reducing bicarbonate ions. Considering that a well developed infrastructure exists for methane transport and storage, as opposed to hydrogen gas, methane is preferred as an energy storage medium.

[009] This background information is provided for the purpose of making known information believed by the applicant to be of possible relevance to the present disclosure. No admission is necessarily intended, nor should be construed, that any of the preceding information constitutes prior art against the present disclosure.

SUMMARY:

[0010] The present disclosure provides an electrochemical process for removal of nitrogen-containing compounds from solutions, such as wastewater. Fuel cells are also discussed, in which some or all of the voltage is provided by electrogenic microorganisms.

[0011] Oxygen is not directly involved in the removal of the nitrogen- containing compound, which allows for the extraction of a portion of the energy contained in said nitrogen-containing compounds.

[0012] Microorganisms living in wastewater, or supplied using an external microbial suspension capable of such a process, use the nitrogen- containing compounds to produce electrical current. This process may be assisted by a power source (electrolysis cell) or a chemical electron acceptor across an electrical circuit (fuel cell).

[0013] Various nitrogen-containing compounds are contemplated within the scope of the present disclosure, and are defined further below. Such compounds may be oxidized completely to nitrogen gas, or incompletely to other oxidized nitrogen compounds such as N₄0, N2O, NO, N2O3, NO2, N20₄, N2O5, N03, N02^", N03^", cyanides such as HCN, or various organic nitrogen oxides. In embodiments where nitrogen-containing compounds are fully oxidized, the production of toxic intermediates is reduced. [0014] The disclosed methods produce either useful chemicals when operated in electrolysis mode or electrical power when operated in fuel cell mode. In either case, the energy efficiency of the process may be significantly increased.

[0015] In embodiments where methane is produced, the energy recaptured from the solution being treated can be stored using existing energy infrastructure to produce liquid fuels. Liquid fuels or their precursors may also be produced from the generated methane. Brittling of metals is also decreased. Aside from its energy uses, methane can also be used as a carbon feedstock for the chemical industry, or as an alternative source of hydrogen . The resulting biogas may also be of higher quality due to the absence of CO2 production in the cathodic reaction . Recycling of CO2 production into methane may also reduce greenhouse gases during the treatment process.

[0016] In one broad aspect, there is provided a method for treating an aqueous solution using an electrolytic cell having a first electrode in electrical communication with a second electrode. The method includes the steps of: disposing the aqueous solution within a reaction zone in the electrolytic cell, with effect that the aqueous solution is in electrical communication with the first electrode; and applying a voltage across the first and second electrodes, with effect that at least one nitrogen-containing compound, of the aqueous solution, is oxidized . The reaction zone less than 100 μιτιοΙ L- l of free oxygen.

[0017] In another broad aspect, there is provided a method for treating an aqueous solution using an electrolytic cell having a first electrode in electrical communication with a second electrode. The method includes the steps of: disposing the aqueous solution within a reaction zone in the electrolytic cell, with effect that the aqueous solution is in electrical communication with the first electrode; and applying a voltage across the first and second electrodes, with effect that at least one nitrogen-containing compound, of the aqueous solution, is oxidized . The reaction zone is an anaerobic environment. [0018] In another broad aspect, there is provided a method of treating wastewater. The method includes a reactive process, including within a reaction zone, disposing an operative mixture, including operative wastewater, in communication with an oxidant, with effect that at least one nitrogen-containing compound, within the operative wastewater, become oxidized. The oxidant includes carbon dioxide.

[0019] In another broad aspect, there is provided a method of treating wastewater. The method includes supplying wastewater to a reaction zone such that wastewater becomes disposed in communication with operative

microorganisms within the reaction zone, with effect that: (i) produced CO2, is obtained, and (ii) one or more nitrogen-containing compounds, within the wastewater, interact with operative CO2, including the produced CO2 and any CO2 within the supplied wastewater, with effect that the one or more nitrogen- containing compounds are oxidized.

[0020] In another broad aspect, there is provided a method of generating electric current within an external circuit of a fuel cell having a first electrode and a second electrode, the first electrode being coupled to the second electrode via the external circuit. The method includes : disposing operative wastewater in electrical communication with the first electrode within a first space of the fuel cell, and disposing an oxidant in electrical

communication with the second electrode within a second space of the fuel cell.

[0021 ] In an embodiment, the reaction zone contains less than 100 μΐτιοΙ L ¹ of free oxygen, less than 80 μΐηοΙ L ¹ of free oxygen, less than 50 μΐηοΙ L ¹ of free oxygen, or less than 30 μΐηοΙ L ¹ of free oxygen. There may also be an absence, or substantial absence, of free oxygen within the reaction zone. The oxygen conditions in the reaction zone may also apply to the electrolytic cell as a whole.

[0022] In an embodiment, the first electrode is an anode and the second electrode is a cathode, which are in electrical communication with one another. A voltage may be applied between the anode and the cathode that is between 10 mV and 2500 mV. The anode potential may be at least +400 mV, +400 mV, between +400 mV and + 550 mV, at least + 500 mV, + 500 mV, + 550 mV, or other suitable voltages.

[0023] In an embodiment, the solution containing the at least one nitrogen-containing compound is wastewater, such as municipal wastewater. The at least one nitrogen-containing compound may be oxidized with effect that gaseous nitrogen is produced.

[0024] In an embodiment, a plurality of microorganisms are disposed within the reaction zone, which may be in suspension or located on the first electrode. Such microorganisms may be derived from ocean sediment, anaerobic digester sludge, or other sources.

[0025] In an embodiment, the cell comprises a first portion and a second portion, wherein the first electrode is disposed within the first portion and the second electrode is disposed within the second portion. An ion- exchange membrane may separate the first and second portions.

[0026] In an embodiment, the cell includes an oxidant at the second electrode, which may be disposed in a second reaction zone. The oxidant may include hydrogen ions that are reduced to hydrogen gas. Alternatively, or in addition, carbon dioxide or bicarbonate may be reduced to an organic compound, such as a hydrocarbon, including methane gas. The resulting hydrogen and/or methane gas may be collected and stored.

[0027] In another broad aspect, there is provided a method of detecting a target material within wastewater. The method includes disposing operative wastewater, including the target material, in electrical communication with a first electrode within a first space of a galvanic cell. An oxidant is disposed in electrical communication with a second electrode within a second space of the galvanic cell. The wastewater, the oxidant, the first electrode, and the second electrode co-operate to generate an electrical current within an external circuit of the galvanic cell. The generated current is sensed and in response to the sensing, a signal is transmitted, representative of the detection of the target material. In an embodiment, a method of treating the wastewater is initiated in response to the signal.

[0028] In another broad aspect, there is provided a method of treating wastewater. The method includes: disposing operative wastewater, including the target material, in electrical communication with a first electrode within a first space of a galvanic cell; and disposing an oxidant in electrical

communication with a second electrode within a second space of the galvanic cell; wherein the wastewater, the oxidant, the first electrode, and the second electrode co-operate to generate an electrical current within an external circuit of the galvanic cell; sensing the generated current; and in response to the sensing, modulating a treatment of the wastewater.

[0029] In an embodiment, the modulating includes initiating the treatment of the wastewater, suspending the treatment of the wastewater, increasing a stimulus applied to the wastewater for the treatment of

wastewater, or decreasing a stimulus applied to the wastewater for the treatment of wastewater.

BRIEF DESCRIPTION OF THE DRAWINGS

[0030] FIG 1 is a Pourbaix diagram showing the relationship between pH and the redox potentials Eh of ammonium oxidation (102), proton reduction (101), and proton reduction at +0.5 volts (103) versus a standard hydrogen electrode.

[0031 ] FIG 2 is a schematic electrolytic cell for carrying out an embodiment, in which ammonium is oxidized while producing hydrogen gas. [0032] FIG 3A depicts nitrogen evolution in an electrolytic embodiment over two fed-batch cycles (304, 305) poised at +0.550 volts and three subsequent cycles (306, 307, 308) poised at +0.4 volts. All voltages are versus a standard hydrogen electrode. The results using microbial innocula derived from three separate locations on the ocean floor (301, 302, 303) are shown.

[0033] FIG 3B depicts hydrogen evolution in an electrolytic embodiment over two fed-batch cycles (304, 305) poised at +0.55 volts and the three subsequent cycles (306, 307, 308) poised at +0.4 volts. All voltages are versus a standard hydrogen electrode. The results using microbial innocula derived from three separate locations on the ocean floor (301, 302, 303) are shown.

[0034] FIG 4 is a Pourbaix diagram showing the relationship between pH and redox potentials Eh of ammonium oxidation (101), proton reduction

(102) , CO2 reduction to methane (401), and proton reduction at +0.5 volts

(103) . All voltages are versus a standard hydrogen electrode.

[0035] FIG 5 is a schematic electrolysis reactor carrying out an embodiment, in which ammonium is oxidized while producing methane gas.

[0036] FIG 6A depicts the % Nitrogen removal in an embodiment using a carbon brush anode (601) or a graphite granule "drum" anode (603).

[0037] FIG 6B depicts the combined evolution of methane on the cathode and the anode in mol/L/day using a carbon brush anode (601) or a graphite granule "drum" anode (603) in an embodiment.

[0038] FIG 6C depicts the evolution of n itrogen gas in mol/L/day at the anode (203) or a cathode (604) of an embodiment. DETAILED DESCRIPTION

[0039] To gain a better understanding, the following illustrative embodiments are set forth . It will be understood that these embodiments are intended to describe examples only and are not intended to limit the scope of the present disclosure in any way.

Definitions

[0040] "microorganisms" includes bacteria archaea, and eukarya of the genera : Acidobacterium, Geothrix, Holophaga, Mycobacterium,

Microbacterium, Marinobacter, Paludibacter, Petrimonas, Proteiniphilum, Sediminibacterium, Anaerolinea, Leptolinea, Caldilinea, Deinococcus, Thermus, Clostridium, Bacillus, Butyribacterium, Sporomusa, Acetobacterium,

Acetogenium, Thermoanaerobacter, Anaerovorax, Desulfosporosinus,

Proteiniborus, Faecalibacterium, Fastidiosipila, Hydrogenoanaerobacterium, Oscillibacter, Phascolarctobacterium, Turicibacter, Nitrospira, Nitrososphaera, Nitrosopumilus, Nitrobacter, Kuenenia, Brocardia, Nitrosomonas,

Nitrosopumilus, Nitrosococcus, Nitrospina, Pirellula, Brevundimonas,

Bradyrhizobium, Hyphomicrobium, Pedomicrobium, Xanthobacter, Methylosinus, Nordella, Rhodobium, Amaricoccus, Rhodobacter, Roseomonas, Botryococcus, Synechococcus, Synechocystis, Chloroflexus, Chlorobium, Sphingobium, Sphingomonas, Sphingopyxis, Chitinimonas, Ralstonia, Comamonas,

Methylibium, Ottowia, Pelomonas, Herbaspirillum, Thiobacillus, Methylobacillus, Neisseria, Gallionella, Nitrosomonas, Azovibrio, Dechloromonas,

Methyloversatilis, Propionivibrio, Kuenania, Thauera, Bdellovibrio,

Desulfobacterium, Desulfobulbus, Desulfovibrio, Desulfuromonas, Geobacter, Anaeromyxobacter, Haliangium, Desulfobacca, Smithella, Syntrophus,

Syntrophobacter, Syntrophorhabdus, Sulfurospirillum, Sulfuricurvum, Wolinella, Aeromonas, Haliea, Citrobacter, Methylobacter, Methylocaldum, Methylomonas, Methylosinus, Acinetobacter, Pseudomonas, Dokdonella, Thermomonas,

Exilispira, Aminiphilus, Kosmotoga, Verrucomicrobia, Opitutus, Puniceicoccus, Tissierella, Sphingopyxis, Pseudoxanthomonas, Sterolibacterium, Brucella, Bosea, Brevundimonas, Singulisphaera, Azospira, Gemmatimonas,

Sphingobacterium, Azonexus, Aquamicrobium, Petrimonas, Fontibacter,

Arcobacter, Chryseobacterium, Megasphera, Truepera, Hydrogenophaga, Paracoccus, Stenotrophomonas, Methanobacterium, Methanobrevibacter, Methanothermobacter, Methanothermus, Methanomicrobium, Methanogenium, Methanoplanus, Methanoplanus, Methanolacinia, Methanocorpusculum,

Methanofollis, Methanolinea, Methanoculleus, Methanosphaerula, Methanolinea, Methanospirillum, Methanoregula, Methanofollis, Methanocalculus,

Methanothrix, Methanosarcina, Methanosaeta, Methanosphaera,

Halomethanococcus, Methanohalobium, Methanosalsum,

Methanomethylovorans, Methanimicrococcus, Methanohalophilus, Methanolobus, Methanococcoides, Methanococcus, Methanoflorens, Methanohalophilus,

Methanopyrus, Halobacterium, Thermococcus, Pyrococcus, Thermoproteus, and Saccharomyces.

[0041] "nitrogen-containing compound" includes various organic and inorganic molecules containing nitrogen groups, such as amines, including without limitation ammonium, ammonium hydroxide, ethyl amine, hydroxyl amine, benzyl amine, and various organic nitrogen compounds including urea, trimethyl amine, ethyl amine, ethanolamine, or natural and artificial amino acids such as alanine, glutamine, arginine, cysteine, and serine. In some

embodiments, the nitrogen-containing compound may also be drugs such as paracetamol, oxetacaine, chlorphenamine, chlorpromazine, amphetamine, clomipramine, or nortriptyline. Other nitrogen-containing compounds may be amides and alkylated amides such as acetamide, formamide, sulfonamide, phosphoramide, N-methylacetamide, or acrylamide. Additional nitrogen- containing compounds include glucosamines, for example glucosamine or N- acetylglucosamine or polymers of N-acetylglucosamine, such as chitin and peptidoglycan. Further nitrogen-containing compounds include nucleotides, deoxynucleotides, and purines or pyrimidines such as adenine, uridine, guanine, cytosine, and thymine. The nitrogen-containing compound may also be a polymer of the foregoing, such as polyamines, polyamides, or polynucleotides (e.g. RNA or DNA). Example 1 : Electrolytic Nitrogen Removal Coupled to Hydrogen Gas Production

[0042] FIG 1 is a Pourbaix diagram showing the relationship between pH on the X-axis and the redox potentials Eh on the Y-axis of ammonium oxidation (101) and proton reduction (102). Also shown is the hypothetical offset potential (103) of proton reduction at +0.5 Volts versus a standard hydrogen electrode.

[0043] Under the standard conditions (all concentrations, except protons, 1 M, gases 1 bar, temperature 25°C) shown in FIG 1, the oxidation of dissolved ammonium 101 (N2/N H4^"1") with protons 102 (Ι- /Η2) as electron acceptor is not spontaneous. This is true for pH values (X-axis) between 0 and 14.

Table 1: Gibbs Free Energy of the complete reactions for oxidation of various nitrogen-containing compounds using protons as the oxidant:

[0044] An electron acceptor more positive than N2/NH4^"1" would ordinarily be needed to make the oxidation of ammonium chemically

spontaenous. However, an applied electrode potential (103), for example set to + 500 mV, can be used instead of a chemical electron acceptor to drive the reaction. At + 500 mV, the oxidation of ammonium to N2 is spontaneous and hydrogen gas can be produced at the cathode.

[0045] FIG 2 is a schematic of the electrolytic cell used in Example 1 to generate hydrogen gas from an aqueous ammonium solution. In th is Example, the cell is divided into a first compartment (206) containing an anode (203) and a second compartment (207) containing a cathode (204). The anode (203) and cathode (204) are partially submerged in an electrolyte (202). The electrolyte (202) also links the two compartments ionically, which were divided by an ion- exchange membrane (210) (Nafion™ 117, Chemours Company, New Castle, Delaware). The anode (203) and cathode (204) were in electrical

communication with one another via a DC power source (208), the electrolyte (202), and the ion-exchange membrane (210). In Example 1, the DC power source (208) was an AC/DC converter, which is a potentiostat. A reference electrode (209) is provided in order to assist the DC power supply (208) in maintaining a consistent voltage.

[0046] Cells of this configuration are described in Siegert et al.

"Comparison of Nonprecious Metal Cathode Materials for Methane Production by Electromethanogenesis" ACS Sustainable Chem. Eng., 2014, 2(4), pp 910-917, which is incorporated herein by reference in its entirety.

[0047] Each of the reactor chambers (Adams & Chittenden, Berkely,

California, USA) was sealed using a screw cap (GL45 Corning Screw Caps, Thomas Scientific, Swedesboro, New Jersey, USA) with a center hole having a septum. The septa were self-made 45 mm discs from 7 mm thick butyl rubber. The two reactor-halves were held together using 35/25 pinch clamps (Thomas Scientific, Swedesboro, New Jersey, USA).

[0048] In other embodiments, the ion-exchange membrane (210) may be omitted. Likewise, the second compartment (207) may further include a collection means for collecting and storing hydrogen gas. Various other DC power sources (208) may also be substituted for the AC/DC converter, such as batteries, solar cells, and the like.

[0049] In Example 1, the anode (203) was a cylindrical carbon fiber brush (4 cm x 4 cm) made from carbonized polyacrylonitrile fibers sold as Panex 35™ (Zoltek in St. Louis, Missouri, USA). Graphite blocks

(2 x 2 x 0.32 cm) were used as cathode (204), which were sanded with 1,500 grit sandpaper and washed without further treatment. The current collector on the cathode (204) was a titanium wire inserted into the graphite block, through drill holes.

[0050] Each electrode was connected to the DC power supply (208) by titanium wires. An Ag/AgCI reference electrode (209) (model RE-5B, BASi, West Lafayette, Indiana, USA) was inserted through holes in rubber septa sealing the top of the first (206) compartment. [0051 ] The solution to be treated in Example 1 was 200 mL of artificial seawater containing 5 mM ammonium chloride and 30 mM sodium bicarbonate. This served as the electrolyte (202), which was added to the first (206) and second (207) compartment. The use of artificial seawater ensured that ammonium was the primary source of electrons and nitrogen in the solution being treated and sodium bicarbonate was the primary carbon source available. A head space of about 50 mL was left in each compartment (206, 207). In alternative embodiments, the electrolye (202) may be municipal wastewater or other nitrogen containing solutions in need of treatment and/or a second electrolyte, such as a suitable buffer, may also be used in the second (207) compartment.

[0052] Trace nutrients, minerals, and other growth media were also added to the artificial seawater to support microbial growth . The resulting artificial seawater solution was as follows :

• 5 mM ammonium chloride

• 10 mL/L of the following trace element solution :

Nitrilotracetic acid 1.5 g/L

MgS0₄ x 7H₂0 3 g/L

NaCI 1 g/L

ZnS0₄ 0.1 g/L

CuS0₄ x 5H₂0 0.01 g/L

AIK(S0₄)₂ 0.01 g/L

• 10 mL/L of the following vitamin solution : Pyridoxin x 2HCI 50 mg/L

Thiamin x 2HCI 10 mg/L

Bi2 (cyanocobalamine) 10 mg/L

p-Aminobenzoic acid 10 mg/L

Riboflavin 5 mg/L

Nicotinic acid 5 mg/L

Ca-D(+)-pantothenate 5 mg/L

Lipoic (thioctic) acid 5 mg/L

D(+)-biotin 2 mg/L

Folic acid 2 mg/L

ch litre of artificial seawater further contained :

NaHCOs 0.25 g (30 mM)

KCI 0.72 g

KBr 0.09 g

NaCI 26 g

[0053] Further details are provided in Siegert M, Sitte J, Galushko A,

Kruger M (2014a), "Starting up microbial enhanced oil recovery." In : Schippers A, Glombitza F, Sand W (eds) Geobiotechnology II. Springer Berlin Heidelberg, pp 1-94, which is incorporated herein by reference in its entirety.

[0054] The first compartment (206) of the electrolytic cell was also inoculated with ocean floor sediment from one of three different locations in the Atlantic Ocean, off the coast of Namibia . Three separate inocula (301, 302, 303) were collected from ocean sediments using gravity corers. The first inoculum (301) was collected at 25°45.060S and 13°04.200E at a water depth of 1,942 m and a sediment depth of 308 cm. The second inoculum (302) was collected at 26°22.178S and 11°53.492E in 3795.6 m water and 0-431 cm sediment depth . The third inoculum (303) was collected at 27°44.131S and 14°14.553E in 1249.3 water and 8-88 cm sediment depth. Live cultures were taken using 1 mL of gravity core sediment, diluted 1/5 with on-site seawater and stored over several years. One mL of the 1/5 (i.e. 20%) dilution was used to inoculate the first compartment (206). Without committing to a particular theory, it is believed that this mixed ocean floor inoculum contained a variety of microorganisms from various genera, including electrogenic species.

[0055] In operation, the power supply (208) was adjusted to maintain

+400 mV or + 550 mV at the anode (203), depending on the treatment.

Potentials reported here are expressed versus a standard hydrogen electrode (SHE), which has an approximate offset potential to an Ag/AgCI electrode of about +0.2 volts.

[0056] The cells were operated in fed-batch cycles in which the anode

(203) was poised at + 550 mV during the first two cycles (304, 305) and at +400 mV during the last three cycles (306, 307, 308) using a potentiostat as the power source (208). All voltages are versus a standard hydrogen electrode.

[0057] While both compartments were batch-fed, only the first compartment (206) was inoculated with microorganisms. Re-inoculation of the first compartment (206) was carried out with a 10% volume of the solution from the previous batch cycle, resulting a 10-fold dilution series of the existing microorganisms in the first compartment (206). That is, at the end of one fed- batch cycle, 90 mL of solution from the first compartment (206) was discarded. The remaining 10 mL were mixed with 90 mL fresh solution to create a 1 : 9 mix, which was used to fill the first comparment (206) and the fed batch cycle was started anew. In contrast, the second compartment (207) was completely replenished with fresh electrolyte (202) on each feeding.

[0058] Prior to the start of each fed-batch cycle, the headspace in each compartment (206, 207) was flushed using argon gas for at least 5 minutes. This created a substantially anaerobic environment in the electrolytic cell and allowed for more accurate measurements of nitrogen gas production. The anodic potentials used (i.e. +400 mV to+ 500 mV) were also too low to cause formation of secondary O2 within the cell through electrolysis of water, which can occur at higher potentials (i.e. +820 mV with Pt catalyst).

[0059] Each batch cycle was operated at room temperature (i.e.

between 20-30 ° C) and standard atmospheric pressure. The total length of the experiment was 600 days.

[0060] FIG 3A depicts nitrogen evolution in the electrolytic cell over five fed-batch cycles (304, 305, 306, 307, 308) with ammonium as the only source of electrons and nitrogen . The results for three electrolytic cells are shown. Each cell was inoculated with one of the three ocean floor sediments (301, 302, 303).

[0061 ] FIG 3B depicts hydrogen evolution in the electrolytic cell over five fed-batch cycles (304, 305, 306, 307, 308) with ammonium as the only source of electrons and nitrogen. The results for three electrolytic cells are shown. Each cell (301, 302, 303) was inoculated with one of the three ocean floor sediments (301, 302, 303).

[0062] The above experiment was repeated by the inventor, with similar results.

[0063] The predicted half reactions are as follows :

6 FT + 6 e-→ 3 H₂

[0064] Without necessarily committing to a particular theory, it is believed that the microorganisms formed a biofilm on the anode (203). In a first reaction zone, these microorganisms transferred electrons from the oxidation reaction (201) into the anode (203), which passed to the cathode (204) via the power supply (208) under the applied voltage. Simultaneously, hydrogen ions generated during the oxidation reaction (101) in the first reaction zone migrated across the membrane (210), and into the second compartment (207). In the second compartment (207), electrons arriving from the cathode (204) and hydrogen ions arriving from the first compartment (206) are consumed in a reduction reaction (205) at a second reaction zone around the cathode (204).

Example 2 : Electrolytic Denitrification Coupled to Methane Gas Production

[0065] FIG 4 is a Pourbaix diagram showing the relationship between pH and redox potentials Eh of ammonium oxidation (101), proton reduction (102), and CO2 reduction to methane (401). Also shown is the hypothetical offset potential of proton reduction (103) at +0.5 volts, for comparison.

Table 2: Gibbs Free Energy for complete reactions for the oxidation of various nitrogen-containing compounds using carbon dioxide as the oxidant:

Δσ°' kj

Net reaction

mol^{" 1}

Ammonium

8 NH₄ ⁺ + 3 HCO3- → 4 N2 + 3 CH₄ + 5 H⁺ + 9 H2O -90

3 NH₄ ⁺ + HCO3- → IN3- + CH₄ + 3 H⁺ + 3 H2O + 271

2 NH₄ ⁺ + HCO3- → N2O + CH₄ + H⁺ + 2 H2O + 284

8 NH₄ ⁺ + 5 HCO3- → 8 NO + 5 CH₄ + 3 H⁺ + 7 H2O + 2,237

4 NH₄ ⁺ + 3 HCO3- → 4 NO2- + 3 CH₄ + 5 H⁺ + H2O + 1,341

4 NH₄ ⁺ + 3 HCO3- → 2 N2O3 + 3 CH₄ + H⁺ + 3 H2O + 1,459

8 NH₄ ⁺ + 7 HCO3- → 4 N₂0₄ + 7 CH₄ + H⁺ + 5 H2O + 3,561

8 NH₄ ⁺ + 7 HCO3- → 8 NO2 + 7 CH₄ + H⁺ + 5 H2O + 3,572

NH₄ ⁺ + HCO3- → NO3- + CH₄ + H ⁺ + 464

2 NH₄ ⁺ + 2 HCO3- → N2O5 + 2 CH₄ + H2O + 1, 111

Ethyl Amine

[0066] As seen in FIG 4 and Table 2, the oxidation of ammonium using carbon dioxide as the oxidant proceeds spontaneously (AG°' = -90 kJ mol^"1), as follows :

8 NH₄ ⁺ + 3 HCO3-→ 3 CH4 + 4 N2 + 5 H⁺ + 9 H2O

[0067] Accordingly, one advantage of producing methane gas is that a power supply is not always required to drive the reaction. Nevertheless, the application of a potential can increase the kinetics of the reaction .

[0068] FIG 5 is a schematic of the electrolytic cell used in Example 2, which is substantially the same construction as in Example 1. However, in this example, the ion-exchange membrane (210) was omitted so as to allow free movement of microorganisms between the first (206) and second (207) compartments. [0069] In other embodiments, the ion-exchange membrane (210) may be included, as shown in FIG 2. Likewise, the second compartment (207) may further include a collection means for collecting and storing methane.

[0070] Two configurations were tested for the anode (203), with three replicate electrolytic cells in each configuration. In the first configuration, cylindrical carbon fiber brushes (4 cm x 4 cm) made from carbonized polyacrylonitrile fibers sold as Panex 35™ (Zoltek in St. Louis, MO, USA) were used as the anode (203). In the second configuration, a "drum" style anode (601) was used, which comprised a cylindrical titanium mesh basket (4 cm x 4 cm, mesh size 40, Ti wire as current collector) with a closed top and bottom, in which the basket was filled with untreated graphite granules of 0.5-5 mm diameter and irregular shapes. Electrodes of this type are described by Siegert in PCT application No. PCT/IB2018/052671, filed April 2018, which is

incorporated by reference herein. The cathodes (504) were untreated 4 cm x 4 cm stainless steel brushes (type 304) purchased from Gordon Brush in

Commerce, CA, USA.

[0071 ] In operation, the power supply (208) was adjusted to maintain

+ 500 mV at the anode (203). Potentials reported here are expressed versus a standard hydrogen electrode (SHE), which has an approximate offset potential to an Ag/AgCI electrode of about +0.2 volts.

[0072] The solution to be treated was domestic wastewater collected at

Calgary's Fish Creek Wastewater Treatment Plant, usually during the morning hours. Pure wastewater was obtained from the primary clarifier. Total Kjeldah Nitrogen ("TKN") concentrations varied from 43 to 120 mg/L (N) or 3-9 mM.

[0073] In Example 2, the microorganisms were provided endogenously within the wastewater, which was enriched by a 1% inoculum of anaerobic digester sludge obtained from the same wastewater treatment plant. This wastewater solution was supplied to the first (206) and second (207) compartments of the electrolytic cell. In alternative embodiments, a different electrolyte, such as a suitable buffer, may instead be used in the second (207) compartment, in place of wastewater.

[0074] Without committing to a particular theory, it is believed that the inoculum derived from the wastewater and anaerobic digester sludge contained a variety of microorganisms from various genera, including electrogenic and methanogenic species.

[0075] Once the experiment was underway, no additional inoculum was used from the anaerobic digester. Instead, the electrolytic cells were batch fed using a 10% volume of solution from the previous batch cycle mixed with freshly collected wastewater, resulting a 10-fold dilution series. Each fed-batch cycle lasted until gas evolution ceased but no longer than ten days. Each batch cycle was operated at room temperature (i.e. between 20-30°C) and standard atmospheric pressure.

[0076] Each cell was operated as a closed system, such that oxidative decomposition of organic and other materials in the wastewater quickly created an anaerobic environment within the cells. The anodic potentials used (i.e. 0.5 volts) were also too low to cause formation of secondary O2 within the cell through electrolysis of water, which can occur at higher potentials (i.e. +0.82 volts with a Pt catalyst). The average O2 concentration within the wastewater was 42± 34 μΜ with the error being the standard deviation, n = 392

measurements. On average, there was a slight increase in O2 levels within the head space (1 μΜ / day) due to sampling and leakage. Nevertheless, O2 concentrations within the wastewater continued to decrease over the course of the experiment. The maximum observed O2 concentration within the wastewater was approximately 80 μΜ at one measurement. O2 concentrations of less than 100 μΜ, less than 50 μΜ, and less than 30 μΜ are also

contemplated within the scope of the present disclosure.

[0077] In Example 2, the oxidant was carbon dioxide, which was reduced to methane. The CO2 in the cathode reaction (205) may be in the form of bicarbonate ions within the wastewater to be treated (e.g. due to the previous oxidation of organic molecules in the wastewater) or can be supplied to the electrolytic cell by an outside source, such as CO2 sparging or gaseous supply to the head space in the second compartment (207).

[0078] FIG 6A depicts the % nitrogen removal using a carbon brush anode (603) or a graphite granule "drum" anode (601). Total nitrogen was determined using the TKN method.

[0079] FIG 6B depicts the combined evolution of methane from both compartments (206, 207) in mol/L/day using a carbon brush anode (603) or a graphite granule "drum" anode (601) of the type described above. The cathodes were in all cases the steel brushes described above. Gases were measured using a gas chromatograph. During the first four fed-batch cycles, the second compartment (207) also produced hydrogen gas (not shown) which was consumed by methanogenic microorganisms and converted into methane gas. During the last fed-batch cycle, hydrogen gas accumulated only during the first day.

[0080] FIG 6C depicts the evolution of n itrogen gas in mol/L/day at the anode (601) and cathode (604) of the electrolytic cells. A graphite granule "drum" style anode (601) was used in this experiment, which was performed in triplicate. As seen in FIG 6C, there was more dinitrogen gas in the anode compartment than in the cathode compartment indicating an oxidative process, which in the case of wastewater can be the oxidation of organic nitrogen or ammonium .

[0081 ] The predicted half reactions are as follows :

CO2 + 8 e^" + 8 FT→ CH4 + 2 H2O [0082] Without necessarily committing to a particular theory, it is believed that microorganisms populated both the anode (203) and the cathode (504) of the electrolytic cell. Electrons and hydrogen ions were transferred from the oxidation reaction (201) in a first reaction zone around the anode (203) to a second reaction zone around the cathode (504), in a similar manner to Example 1 above. In Example 2, microorganisms were also present in the second compartment (207), possibly as a biofilm on the cathode (504). It is believed that methanogens amongst the population of micoorganisms were responsible for the reduction reaction (401), in which incoming hydrogen ions from the electrolyte (202) were combined with incoming electrons from the cathode (504) in the presence of bicarbonate, at a second reaction zone in the second compartment (207), to produce methane and water. Intermediates in th is process may include hydrogen gas, which is likely consumed by the

methanogens when producing methane.

Example 3 : Fuel Cell Nitrogen Removal

[0083] Referring to Figure 7, in another aspect, there is provided a fuel cell (700). The fuel cell (700) includes an anode compartment (702) and a cathode compartment (704). The anode compartment (702) is separated from the cathode compartment (704) by a separator (706) configured to permit selective permeation of ionic species therethrough. An anode (708) is disposed within the anode compartment (702) and a cathode (710) is disposed within the cathode compartment (704). The anode (708) and cathode (710) are electrically coupled to an external load (712) via an external circuit (714). A reference electrode (713) may be used to monitor the redox potential at either of the electrodes.

[0084] Although control over the voltage may be more difficult, a fuel cell (700) is more efficient as it avoids two energy conversion steps typical of electrolytic arrangements, namely: (a) conversion of electricity into chemical energy; and (b) conversion of chemical energy back into electricity. [0085] Wastewater is supplied to the anode compartment (702) such that the wastewater becomes disposed within the anode compartment (702) in electrical communication with the anode (708). In some embodiments, for example, the wastewater includes nitrogen-containing or organic compounds that serve as the energy source for the fuel cell.

[0086] Oxidant is supplied to the cathode compartment (704) such that the oxidant becomes disposed within the cathode compartment (704) in electrical communication with the cathode (710). Such an oxidant may be gaseous oxygen dissolved in solution but can be any other oxidizing compound. In some embodiments, for example, the oxidant includes02, benzoyl peroxide, ferricyanide, manganese dioxide, or nitrate.

[0087] Mediated by electrical communication, the oxidant reacts with a fuel source to create a cathodic reduction reaction with a redox potential more positive than the anodic oxidation reaction . Example compounds that may serve as the energy source for the fuel cell include nitrogen-containing compounds and organic compounds, which may be found in wastewater or may be supplied separately to the cathode compartment (704). Examples fuel sources include ammonium, benzylamine, ethanolamine, arginine, cysteine, serine, acetamide, N-methylacetamide, or other nitrogen-containing

compounds.

[0088] An example of such a reaction mediated by the fuel cell (700) is the oxidation of ammonium in the presence of molecular oxygen (2 NhU^"1" + 3 O2 → 2 N₂ + 4 H⁺ + 6 H2O; AG°' = - 1,265 kJ mol^"1). Various other nitrogen- containing compounds or organic molecules may be similarly oxidized to provide the required current.

[0089] While: (i) the wastewater is disposed in electrical communication with the anode (708), and (ii) the oxidant is disposed in electrical

communication with the cathode (710), electrical current is generated across the external circuit (714) of the fuel cell (700). Such current may be used to drive the anodic reaction and/or be used to power a useful external load (712) such as a resistor or battery.

[0090] In some embodiments, a plurality of microorganisms may be disposed within the anode compartment (702) to assist the anodic reaction. In some embodiments, for example, at least a portion of such microorganisms may be coated on the anode (708) as a biofilm. These microorganisms may assist in nitrogen removal in the manner described in Examples 1 and 2 above.

[0091 ] In yet another aspect, the electrochemical cell in Example 2

(see: FIG 5) may itself be configured as a microbial fuel cell. As discussed, the oxidation of ammonium using carbon dioxide under anaerobic conditions is a spontaneous reaction (AG°' = -90 kJ mol^"1). In this embodiment, the power supply (208) is replaced by a useful electrical circuit, and the voltage potential is generated by spontaneous oxidation of nitrogen-containing compounds by electrogenic microorganisms at the anode (203).

[0092] In yet another aspect, the fuel cell (700) can be used for detecting target material within the wastewater, and the load (712) is an electrical current sensor. In this respect, in some embodiments, for example, the disposition of the wastewater within the fuel cell (700) is with effect that an electrical current is generated that is representative of the presence of a target material within the wastewater. The sensing of the current by the sensor can be used to indicate detection of the target material within the wastewater. In response to the sensing, a signal can be transmitted, representative of the detection of the target material. Examples of such targets include nitrogen- containing compounds.

[0093] The detection of nitrogen-containing compounds may be used as a signal to begin treating the wastewater, suspending the treatment of the wastewater, increasing a stimulus applied to the wastewater for the treatment of wastewater, or decreasing a stimulus applied to the wastewater for the treatment of wastewater. The stimulus may include admixing the wastewater with a reagent, or disposing the wastewater within a reaction zone in the electrolytic cell, with effect that the aqueous solution is in electrical

communication with an anode and the anode is disposed in electrical communication with a cathode and applying a voltage, and includes

embodiments discussed herein.

[0094] The embodiments of the present disclosure are intended to be examples only. Those of skill in the art may effect alterations, modifications and variations to the particular embodiments without departing from the intended scope of the present application.

[0095] In particular, features from one or more of the above-described embodiments may be selected to create alternate embodiments comprised of a subcombination of features which may not be explicitly described above. In addition, features from one or more of the above-described embodiments may be selected and combined to create alternate embodiments comprised of a combination of features which may not be explicitly described above. Features suitable for such combinations and subcombinations would be readily apparent to persons skilled in the art upon review of the present application as a whole. The subject matter described herein and in the recited claims intends to cover and embrace all suitable changes in technology.

Claims

CLAIMS:

1. A method for treating an aqueous solution using an electrolytic cell

having a first electrode in electrical communication with a second electrode, the method comprising : disposing the aqueous solution within a zone in the electrolytic cell, with effect that the aqueous solution is in electrical communication with the first electrode; and applying a voltage across the first and second electrodes, with effect that at least one nitrogen-containing compound, of the aqueous solution, is oxidized; and wherein the zone contains less than 100 μΐηοΙ L ¹ of free oxygen.

2. A method for treating an aqueous solution using an electrolytic cell

having a first electrode in electrical communication with a second electrode, the method comprising : disposing the aqueous solution within a zone in the electrolytic cell, with effect that the aqueous solution is in electrical communication with the first electrode; and applying a voltage across the first and second electrodes, with effect that at least one nitrogen-containing compound, of the aqueous solution, is oxidized; and wherein the zone is an anaerobic environment.

3. The method of claim 1 or claim 2, wherein the zone contains less than 80 μΐτιοΙ L ¹ of free oxygen, such as less than 50 μΐηοΙ L ¹ of free oxygen, such as less than 30 μΐηοΙ L ¹ of free oxygen .

4. The method of any one of claims 1 to 3, wherein there is an absence, or substantial absence, of free oxygen within the zone.

5. The method of any one of claims 1 to 4, wherein the electrolytic cell is an anaerobic environment.

6. The method of any one of claims 1 to 5, wherein there is an absence, or substantial absence, of free oxygen within the electrolytic cell.

7. The method of any one of claims 1 to 6, wherein the at least one

nitrogen-containing compound is oxidized in the absence, or substantial absence, of free oxygen.

8. The method of any one of claims 1 to 7, wherein the free oxygen is O2.

9. The method of any one of claims 1 to 8, wherein the first electrode is disposed within the zone.

10. The method of any one of claims 1 to 9, wherein the first electrode is an anode and the second electrode is a cathode.

11. The method of any one of claims 1 to 10, wherein the voltage is at least +400 mV, at least + 500 mV, or between+400 mV and + 550 mV.

12. The method of any one of claims 1 to 11, wherein the oxidation of the at least one nitrogen-containing compound is with effect that gaseous nitrogen is produced in the first space.

13. The method of claim 12, wherein at least 0.01 weight % of the nitrogen- containing compound disposed within the zone is oxidized into nitrogen gas, such as at least 1.0 weight %, such as at least 10 weight %, and such as at least 50 weight %.

14. The method of any one of claims 1 to 13, wherein a plurality of

microorganisms are disposed within the zone.

15. The method of claim 14, wherein the plurality of microorganisms form a biofilm on the first electrode. The method of claim 14 or claim 15, wherein the microorganisms are derived from ocean floor sediment or anaerobic digester sludge.

The method of any one of claims 14 to 16, wherein the plurality of microorganisms are selected from the genera consisting of:

Acidobacterium, Geothrix, Holophaga, Mycobacterium, Microbacterium, Marinobacter, Paludibacter, Petrimonas, Proteiniphilum,

Sediminibacterium, Anaerolinea, Leptolinea, Caldilinea, Deinococcus, Thermus, Clostridium, Bacillus, Butyribacterium, Sporomusa,

Acetobacterium, Acetogenium, Thermoanaerobacter, Anaerovorax, Desulfosporosinus, Proteiniborus, Faecalibacterium, Fastidiosipila, Hydrogenoanaerobacterium, Oscillibacter, Phascolarctobacterium, Turicibacter, Nitrospira, Nitrososphaera, Nitrosopumilus, Nitrobacter, Kuenenia, Brocardia, Nitrosomonas, Nitrosopumilus, Nitrosococcus, Nitrospina, Pirellula, Brevundimonas, Bradyrhizobium, Hyphomicrobium, Pedomicrobium, Xanthobacter, Methylosinus, Nordella, Rhodobium, Amaricoccus, Rhodobacter, Roseomonas, Botryococcus, Synechococcus, Synechocystis, Chloroflexus, Chlorobium, Sphingobium, Sphingomonas, Sphingopyxis, Chitinimonas, Ralstonia, Comamonas, Methylibium, Ottowia, Pelomonas, Herbaspirillum, Thiobacillus, Methylobacillus, Neisseria, Gallionella, Nitrosomonas, Azovibrio, Dechloromonas,

Methyloversatilis, Propionivibrio, Kuenania, Thauera, Bdellovibrio, Desulfobacterium, Desulfobulbus, Desulfovibrio, Desulfuromonas, Geobacter, Anaeromyxobacter, Haliangium, Desulfobacca, Smithella, Syntrophus, Syntrophobacter, Syntrophorhabdus, Sulfurospirillum, Sulfuricurvum, Wolinella, Aeromonas, Haliea, Citrobacter, Methylobacter, Methylocaldum, Methylomonas, Methylosinus, Acinetobacter,

Pseudomonas, Dokdonella, Thermomonas, Exilispira, Aminiphilus, Kosmotoga, Verrucomicrobia, Opitutus, Puniceicoccus, Tissierella, Sphingopyxis, Pseudoxanthomonas, Sterolibacterium, Brucella, Bosea, Brevundimonas, Singulisphaera, Azospira, Gemmatimonas,

Sphingobacterium, Azonexus, Aquamicrobium, Petrimonas, Fontibacter, Arcobacter, Chryseobacterium, Megasphera, Truepera, Hydrogenophaga, Paracoccus, Stenotrophomonas, Methanobacterium, Methanobrevibacter, Methanothermobacter, Methanothermus, Methanomicrobium,

Methanogenium, Methanoplanus, Methanoplanus, Methanolacinia, Methanocorpusculum, Methanofollis, Methanolinea, Methanoculleus, Methanosphaerula, Methanolinea, Methanospirillum, Methanoregula, Methanofollis, Methanocalculus, Methanothrix, Methanosarcina,

Methanosaeta, Methanosphaera, Halomethanococcus, Methanohalobium, Methanosalsum, Methanomethylovorans, Methanimicrococcus,

Methanohalophilus, Methanolobus, Methanococcoides, Methanococcus, Methanoflorens, Methanohalophilus, Methanopyrus, Halobacterium, Thermococcus, Pyrococcus, Thermoproteus, and Saccharomyces.

18. The method of any one of claims 14 to 17, wherein the aqueous solution further comprises nutrients and buffers to support growth of the plurality of microorganisms.

19. The method of claim 18, wherein the step of disposing the aqueous

solution, after a predetermined period of time: sampling a portion of the aqueous solution within the electrolytic cell to create an inoculum; mixing the inoculum with a fresh batch of the aqueous solution; replacing the aqueous solution in the electrolytic cell with the fresh batch .

20. The method of any one of claims 1 to 19, wherein the electrolytic cell comprises a first portion and a second portion, wherein the first electrode is disposed within the first portion and the second electrode is disposed within the second portion.

The method of claim 20, wherein the first portion and the second portio of the electrolytic cell are separated by an ion-exchange membrane.

22. The method of claim 21, wherein the electrolytic cell further comprises an electrolyte disposed in electrical communication with the second electrode.

23. The method of claim 22, wherein the electrolyte contains an oxidant. 24. The method of claim 23, wherein the oxidant is H⁺, which is reduced to hydrogen gas.

25. The method of claim 24, wherein the nitrogen-containing compound

comprises ammonium and three moles of hydrogen gas is generated for every two moles of ammonium oxidized. 26. The method of claim 24 or 25, wherein the nitrogen-containing compound comprises ethylamine or an amino acid and 12 to 15 moles of hydrogen gas are generated for every two moles of ethylamine or amino acid oxidized.

27. The method of any one of claims 14 to 21, wherein the method further comprises the step of: disposing the aqueous solution within a second zone in the electrolytic cell, with effect that the aqueous solution becomes disposed in electrical communication with the second electrode; and applying the voltage across the first and second electrodes, with effect that bicarbonate within the aqueous solution is reduced to an organic compound.

28. The method of claim 27, wherein the organic compound is a hydrocarbon.

29. The method of claim 28, wherein the hydrocarbon is methane gas.

30. The method of any one of claims 27 to 29, wherein the plurality of

microorganisms is also disposed on the second electrode

31. The method of any one of claims 27 to 30, further comprising supplying carbon dioxide to the aqueous solution or the electrolytic cell.

32. The method of any one of claims 1 to 31, wherein the aqueous solution comprises wastewater.

33. The method of any one of claims 1 to 32, wherein the at least one

nitrogen-containing compound comprises ammonium or a salt thereof.

34. The method of claim 33, wherein four moles of nitrogen gas is generated for every eight moles of ammonium oxidized.

35. The method of any one of claims 1 to 34, wherein the at least one

nitrogen-containing compound comprises ethylamine or an amino acid .

36. The method of claim 35, wherein one mole of nitrogen gas is produced for every 1 to 2 moles of ethylamine or amino acid oxidized.

37. The method of any one of claims 1 to 36, wherein the at least one

nitrogen-containing compound is selected from the group consisting of: amines, including ammonium, ammonium hydroxide, ethyl amine, hydroxyl amine, benzyl amine; organic nitrogen compounds including urea, trimethyl amine, ethyl amine, ethanolamine; natural and artificial amino acids including alanine, glutamine, arginine, cysteine, and serine; drugs including paracetamol, oxetacaine, chlorphenamine,

chlorpromazine, amphetamine, clomipramine, or nortriptyline; amides and alkylated amides including acetamide, formamide,

sulfonamide, phosphoramide, N-methylacetamide, or acrylamide; glucosamines, including glucosamine, N-acetylglucosamine, chitin, and peptidoglycan; nucleotides and deoxynucleotides, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA); purines or pyrimidines including adenine, uridine, guanine, cytosine, and thymine; and polymers of the foregoing, including polyamines, polyamides, or

polynucleotides.

38. The method of any one of claims 1 to 36, wherein the at least one

nitrogen-containing compound comprises ammonium.

39. A method of treating wastewater, comprising : effecting a reactive process, including : within a zone, disposing an operative mixtu re, including operative wastewater, in communication with an oxidant, with effect that at least one nitrogen-containing compound, within the operative wastewater, become oxidized ; wherein the oxidant includes CO2.

40. The method of claim 39, wherein the ratio of moles of nitrogen, within the at least one nitrogen-containing compound of the operative wastewater, to moles of CO2, within the oxidant, is between 8 : 1 and 1 : 1, such as between 4: 1 and 1 : 1, and more such as 8 : 3.

41. The method of claim 39 or 40, further comprising, prior to the effecting of the reactive process : monitoring the operative wastewater for nitrogen-containing

compounds, and based on sensing of the nitrogen-containing compounds, effecting the disposing of the operative mixture within the zone.

42. The method of any one of claims 39 to 41, wherein the operative mixture includes operative microorganisms, which facilitate the oxidation of the at least one nitrogen-containing compound.

43. The method of claim 42, wherein the operative microorganisms are

selected from the genera consisting of: Acidobacterium, Geothrix,

Holophaga, Mycobacterium, Micro bacterium, Marinobacter, Paludibacter, Petrimonas, Proteiniphilum, Sediminibacterium, Anaerolinea, Leptolinea, Caldilinea, Deinococcus, Thermus, Clostridium, Bacillus, Butyribacterium, Sporomusa, Acetobacterium, Acetogenium, Thermoanaerobacter,

Anaerovorax, Desulfosporosinus, Proteiniborus, Faecalibacterium,

Fastidiosipila, Hydrogenoanaerobacterium, Oscillibacter,

Phascolarctobacterium, Turicibacter, Nitrospira, Nitrososphaera,

Nitrosopumilus, Nitrobacter, Kuenenia, Brocardia, Nitrosomonas,

Nitrosopumilus, Nitrosococcus, Nitrospina, Pirellula, Brevundimonas, Bradyrhizobium, Hyphomicrobium, Pedomicrobium, Xanthobacter, Methylosinus, Nordella, Rhodobium, Amaricoccus, Rhodobacter,

Roseomonas, Botryococcus, Synechococcus, Synechocystis, Chloroflexus, Chlorobium, Sphingobium, Sphingomonas, Sphingopyxis, Chitinimonas, Ralstonia, Comamonas, Methylibium, Ottowia, Pelomonas, Herbaspirillum, Thiobacillus, Methylobacillus, Neisseria, Gallionella, Nitrosomonas, Azovibrio, Dechloromonas, Methyloversatilis, Propionivibrio, Kuenania, Thauera, Bdellovibrio, Desulfobacterium, Desulfobulbus, Desulfovibrio, Desulfuromonas, Geobacter, Anaeromyxobacter, Haliangium,

Desulfobacca, Smithella, Syntrophus, Syntrophobacter,

Syntrophorhabdus, Sulfurospirillum, Sulfuricurvum, Wolinella,

Aeromonas, Haliea, Citrobacter, Methylobacter, M ethy local d um ,

Methylomonas, Methylosinus, Acinetobacter, Pseudomonas, Dokdonella, Thermomonas, Exilispira, Aminiphilus, Kosmotoga, Verrucomicrobia, Opitutus, Puniceicoccus, Tissierella, Sphingopyxis, Pseudoxanthomonas, Sterolibacterium, Brucella, Bosea, Brevundimonas, Singulisphaera, Azospira, Gemmatimonas, Sphingobacterium, Azonexus, Aquamicrobium, Petrimonas, Fontibacter, Arcobacter, Chryseobacterium, Megasphera, Truepera, Hydrogenophaga, Paracoccus, Stenotrophomonas,

Methanobacterium, Methanobrevibacter, Methanothermobacter,

Methanothermus, Methanomicrobium, Methanogenium, Methanoplanus, Methanoplanus, Methanolacinia, Methanocorpusculum, Methanofollis, Methanolinea, Methanoculleus, Methanosphaerula, Methanolinea,

Methanospirillum, Methanoregula, Methanofollis, Methanocalculus, Methanothrix, Methanosarcina, Methanosaeta, Methanosphaera,

Halomethanococcus, Methanohalobium, Methanosalsum,

Methanomethylovorans, Methanimicrococcus, Methanohalophilus,

Methanolobus, Methanococcoides, Methanococcus, Methanoflorens, Methanohalophilus, Methanopyrus, Halobacterium, Thermococcus, Pyrococcus, Thermoproteus, and Saccharomyces.

44. The method of claim 42 or 43, wherein the effecting of the reactive

process further includes, prior to the disposing of the operative

wastewater in communication with an oxidant: disposing an operative wastewater precursor in communication with the operative microorganisms, with effect that: (i) at least a portion of the CO2 is obtained, and (ii) the operative wastewater is obtained.

45. The method of claim 44, further comprising, prior to the effecting of the reactive process : monitoring the operative wastewater precursor for nitrogen- containing compounds, and based on sensing of the nitrogen- containing compounds, effecting the disposing of the operative mixture within the zone.

46. The method of any one of claims 39 to 45, wherein the oxidant includes less than 100 μΐηοΙ L ¹ of free oxygen, such as less than 80 μΐηοΙ L ¹ of free oxygen, such as less than 50 μΐηοΙ L ¹ of free oxygen, such as less than 30 μΐηοΙ L ¹ of free oxygen.

47. The method of any one of claims 39 to 46, wherein there is an absence, or a substantial absence, of oxygen within the oxidant.

48. The method of any one of claims 39 to 47, wherein the communication between the operative wastewater and the oxidant includes an electrical communication.

49. The method as claimed in claim 48, wherein : the operative mixture is disposed within an electrolytic cell

including an anode that is electrically coupled to a cathode and a voltage is applied across the anode and cathode; the operative wastewater is disposed in electrical communication with the anode; and the oxidant is disposed in electrical communication with the

cathode.

50. The method as claimed in any one of claims 39 to 47, wherein the

communication between the operative and the oxidant includes a chemical reaction communication .

51. The method as claimed in any one of claims 39 to 50, wherein the at least one nitrogen-containing compound comprises ammonium or a salt thereof.

52. The method of any one of claims 39 to 51, wherein the at least one

sulfonamide, phosphoramide, N-methylacetamide, or acrylamide; glucosamines, including glucosamine, N-acetylglucosamine, chitin, and peptidoglycan; nucleotides and deoxynucleotides, including deoxyribonucleic acid

(DNA) and ribonucleic acid (RNA); purines or pyrimidines including adenine, uridine, guanine, cytosine, and thymine; and polymers of the foregoing, including polyamines, polyamides, or

polynucleotides.

53. The method of any one of claims 39 to 51, wherein the at least one

nitrogen-containing compound comprises trimethyl amine.

54. The method of any one of claims 39 to 51, wherein the at least one

nitrogen-containing compound comprises amino acids.

55. A method of treating wastewater, comprising : supplying wastewater to a zone such that wastewater becomes disposed in communication with operative microorganisms within the zone, with effect that: (i) produced CO2, is obtained, and (ii) one or more nitrogen-containing compounds, within the wastewater, interact with operative CO2, including the produced CO2 and any CO2 within the supplied wastewater, with effect that the one or more nitrogen-containing

compounds are oxidized .

56. The method as claimed in claim 55, wherein the ratio of moles of nitrogen, within the one or more nitrogen-containing compounds of the wastewater, to moles of the operative CO2 is between 8 : 1 and 1 : 1, such as between 4: 1 and 1 : 1, and such as 8 : 3.

57. The method of claim 55 or 56, further comprising, prior to the effecting of the reactive process : monitoring the wastewater for nitrogen-containing compounds, and based on sensing of one or more nitrogen-containing compounds, effecting the supplying of the wastewater to the zone.

58. The method of any one of claims 55 to 57, wherein the supplied

wastewater includes less than 100 μΐηοΙ L ¹ of free oxygen, such as less than 80 μΐηοΙ L ¹ of free oxygen, such as less than 50 μΐηοΙ L ¹ of free oxygen, such as less than 30 μΐηοΙ L ¹ of free oxygen .

59. The method of any one of claims 55 to 58, wherein there is an absence, or a substantial absence, of oxygen within the supplied wastewater.

60. The method of any one of claims 55 to 59, wherein the communication between the wastewater and the operative CO2 includes an electrical communication.

61. The method of claim 60, wherein : the operative mixture is disposed within an electrolytic cell

including an anode that is electrically coupled to a cathode and a voltage is applied across the anode and cathode; the wastewater is disposed in electrical communication with the anode; and the operative CO2 is disposed in electrical communication with the cathode.

62. The method of claim 61, wherein the anode is coated with the operative microorganisms.

63. The method as claimed in any one of claims 55 to 62, wherein the

64. The method of any one of claims 55 to 63, wherein the at least one

nitrogen-containing compound comprises ammonium or a salt thereof.

65. The method of any one of claims 55 to 64, wherein the at least one

nitrogen-containing compound comprises ethylamine.

66. The method of any one of claims 55 to 63, wherein the at least one

polynucleotides.

67. The method of any one of claims 55 to 65, wherein the at least one

nitrogen-containing compound comprises amino acids.

68. The method of any one of claims 53 to 65, wherein wherein the operative microorganisms are selected from the genera consisting of:

Pseudomonas, Dokdonella, Thermomonas, Exilispira, Aminiphilus, Kosmotoga, Verrucomicrobia, Opitutus, Puniceicoccus, Tissierella,

Sphingopyxis, Pseudoxanthomonas, Sterolibacterium, Brucella, Bosea, Brevundimonas, Singulisphaera, Azospira, Gemmatimonas,

Methanogenium, Methanoplanus, Methanoplanus, Methanolacinia,

Methanocorpusculum, Methanofollis, Methanolinea, Methanoculleus, Methanosphaerula, Methanolinea, Methanospirillum, Methanoregula, Methanofollis, Methanocalculus, Methanothrix, Methanosarcina,

69. A method for generating electric current within an external circuit of a fuel cell having a first electrode and a second electrode, the first electrode being coupled to the second electrode via the external circuit, the method comprising : disposing operative wastewater in electrical communication with the first electrode within a first space of the fuel cell, and disposing an oxidant in electrical communication with the second electrode within a second space of the fuel cell

70. A method of detecting a target material within wastewater comprising : disposing operative wastewater, including the target material, in electrical communication with a first electrode within a first space of a galvanic cell; disposing an oxidant in electrical communication with a second electrode within a second space of the galvanic cell; wherein the wastewater, the oxidant, the first electrode, and the second electrode co-operate to generate an electrical current within an external circuit of the galvanic cell; sensing the generated current; and in response to the sensing, transmitting a signal, representative of the detection of the target material.

71. The method of claim 69 or 70, wherein : the first electrode is an anode; and the second electrode is a cathode.

72. The method of any one of claims 69 to 71, wherein the oxidant includes

73. The method of any one of claims 69 to 71, wherein the oxidant comprises at least one of O2, benzoyl peroxide, ferricyanide, manganese dioxide, and nitrate.

74. The method of any one of claims 69 to 73, wherein the zone further

contains a plurality of microorganisms.

75. The method of claim 74, wherein the plurality of microorganisms is

coated on the first electrode.

76. The method of any one of claims 69 to 75, wherein wherein the operative microorganisms are selected from the genera consisting of:

77. The method of any one of claims 69 to 76, wherein the target material includes at least one nitrogen-containing compound.

78. The method of claim 77, wherein the at least one nitrogen-containing compound comprises ammonium or a salt thereof.

79. The method of claim 76 or 78, wherein the at least one nitrogen- containing compound comprises ethylamine.

80. The method of claim 77, wherein wherein the at least one nitrogen- containing compound is selected from the group consisting of: amines, including ammonium, ammonium hydroxide, ethyl amine, hydroxyl amine, benzyl amine; organic nitrogen compounds including urea, trimethyl amine, ethyl amine, ethanolamine; natural and artificial amino acids including alanine, glutamine, arginine, cysteine, and serine; drugs including paracetamol, oxetacaine, chlorphenamine,

polynucleotides.

81. The method of claim 77, wherein the at least one nitrogen-containing compound comprises amino acids.

82. A method of treating wastewater comprising : disposing operative wastewater, including the target material, in

electrical communication with a first electrode within a first space of a galvanic cell; disposing an oxidant in electrical communication with a second

electrode within a second space of the galvanic cell; wherein the wastewater, the oxidant, the first electrode, and the

second electrode co-operate to generate an electrical current with in an external circuit of the galvanic cell; sensing the generated current; and in response to the sensing, modulating a treatment of the wastewater.

83. The method as claimed in claim 82, wherein the modulating includes initiating the treatment of the wastewater, suspending the treatment of the wastewater, increasing a stimulus applied to the wastewater for the treatment of wastewater, or decreasing a stimulus applied to the wastewater for the treatment of wastewater.