Methods for treating wastewater to produce biofertilizer

Abstract

Disclosed herein are systems and methods of wastewater digestion to produce a fertilizer. Some systems may include an anaerobic decomposition vessel configured to produce an anaerobic effluent. Systems may transfer the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel. Systems may transfer the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel. Systems may digest the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8. Systems may filter at least the first or second aerobic effluent or both, where the second aerobic effluent comprises inorganic nitrogen in oxyanion form and has a carbon to nitrogen ratio greater than or equal to 3:1.

Description

METHODS FOR TREATING WASTEWATER TO PRODUCE BIOFERTILIZERCROSS-REFERENCE TO RELATED U.S. APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 654,424, filed May 31 ,2024, the content of which is incorporated by reference in its entirety.Field of the Disclosure

[0002] The present disclosure relates to systems and methods for producing a biofertilizer or a biofertilizer component from agricultural waste and / or wastewater.BACKGROUND

[0003] The production of biofertilizers is an important component in sustainable agriculture, offering an environmentally friendly alternative to synthetic fertilizers. Various methods have been developed to use organic waste to create nutrient-rich solutions that enhance soil fertility and promote plant growth. Composting is one of the most traditional and widely used methods and involves the aerobic decomposition of organic materials such as plant residues, animal manure, and food waste. Over weeks to months, microorganisms break down the waste into humus — a nutrient-rich material. The process usually uses regular turning to maintain adequate oxygen levels and results in a solid biofertilizer that is directly applicable to the soil. Vermicomposting is a similar but a distinct method, which employs worms to consume and decompose organic waste. The worms' excretions, known as vermicast, are an exceptionally nutrient-rich form of biofertilizer. This method is generally faster than traditional composting and can be adapted to various scales.

[0004] Anaerobic digestion represents another approach, wherein organic material is broken down in the absence of oxygen, typically in a sealed digester. This process not only produces a digestate rich in nutrients like nitrogen, phosphorus, and potassium but also generates biogas, a renewable energy source. Conversely, Bokashi fermentation utilizes specific anaerobic microorganisms to ferment organic waste quickly, handling types of wastenot easily composted, such as meat and dairy. The fermented material is then buried in the soil to finish decomposing, offering a rapid nutrient boost.

[0005] Liquid biofertilizers are produced through fermentation processes involving microbial cultures that fix atmospheric nitrogen, solubilize phosphorus, or mobilize other essential nutrients. These fertilizers are quick-acting and easily applied but generally have a shorter shelf life than their solid counterparts. On a larger scale, industrial production of biofertilizers may employ advanced biotechnological methods to cultivate specific microorganism strains in controlled environments. Storage and transportation, particularly of liquid biofertilizers, pose challenges due to their short shelf life and susceptibility to contamination. Moreover, effectively integrating biofertilizers into existing farming practices necessitates greater education and adaptation of conventional farming practices.

[0006] Some existing methods for integrating biofertilizers include various irrigation techniques like deep water culture, Nutrient Film Techniques, and Aquaponics. These methods often require clear water with very low carbon content, which is difficult to maintain when using organic fertilizers due to their decomposition requirements and resultant microbial growth. Additionally, these systems are not designed to handle the low concentration and high volume of liquid composting products, leading to operational inefficiencies and potential clogging. The need for continuous and extensive sanitization, especially in systems where sanitizers cannot be used, further complicates the operation, necessitating a constant addition of oxygen to maintain a high oxygen saturation for microbial balance.

[0007] Existing methods fail to provide solutions to the following interconnected issues when transitioning to organic nutrition in irrigation systems. A first challenge for organic fertilizers is to produce nutrition that is available for plants, and that is not otherwise bound up in inaccessible forms. Organic fertilizers themselves are generally not available to the plant as nutrition until microorganisms decompose the material into inorganic salts, which can then be assimilated by the plant. A second challenge concerns the fact that most organic fertilizers begin with around 10% usable nutrition, of which at least 30% will often be lost to feeding microorganisms that decompose the organic fertilizer. This might incentivize using large amounts of material, but biodigesters used to decompose the material are sensitive to overloading and have limits to the ultimate concentration of dissolved solids (nutritional salts)that ultimately can be in solution. Accordingly, the final concentration of digestate for most biodigesters is low and therefore relatively large volumes are added to irrigation systems.

[0008] At present, most conventional irrigation systems are designed to inject perhaps a couple of liters of concentrated nutrients per day, where these organic fertilizer digestates would be added in tens of thousands of gallons per day to keep up. This, in turn, raises issues with maintaining water quality. The output of liquid composting is a large proportion of microbial cells, detritus, and humis. If added to an irrigation system, these suspended solids can cause substantial filter and irrigation tube clogging. It can be cost- prohibitive to sanitize the water with oxidizers since remaining suspended solids and soluble carbon will absorb a large proportion of sanitizer before effective sanitizer levels can become stable in solution. An alternative is to provide a small proportion of carbonaceous feed to maintain live microbial cultures in solution, such as with beneficial "Probiotics". However, such systems must host those organisms and constantly inject liquid or concentrated oxygen to maintain high oxygen saturation.

[0009] As such, there remains a need to develop systems and methods, which provide a liquid compost that is well decomposed, such that the liquid compost would introduce little substrate to feed bacteria once in an irrigation system.SUMMARY OF THE DISCLOSURE

[0010] Some aspects of the disclosure relate to systems and methods for producing a biofertilizer and biofertilizer components from agricultural waste and / or wastewater. Preferably, aspects of the disclosure are directed to systems and methods for producing an organic fertilizer compatible with irrigation systems, aimed at addressing microbial fouling challenges. In some embodiments, a process may begin by converting biological feedstock into a digestate via microbial mineralization. In some embodiments, the digestate is a product rich in inorganic nitrogen and low in carbonaceous biological oxygen demand. In some embodiments, mechanical filtration is used to remove suspended solids from a digestate. In some embodiments, a digestate is processed through dewatering, yielding a concentrate and diluted water.

[0011] An aspect of the disclosure relates to systems and methods for processing input materials to generate an output from a bioreactor by using microorganisms. In someembodiments, one or more outputs of a bioreactor is compatible with the addition of stable carbonaceous supplements, such as Humic or Fulvic acids. In some embodiments, an output of one or more bioreactor is introduced into recirculating irrigation systems, facilitating the maintenance of dissolved solids in the circulating fluid.

[0012] An aspect of the disclosure is directed toward using bacteria for specific functions throughout a decomposition process. In some embodiments, one or more reactors including bacterial strains, or mixtures thereof, may focus on the production of fertilizers that yield nitrate and other minerals from complex feeds while also minimizing waste to microbial biomass. This process may include at least one of the following steps or stages to efficiently produce a desired liquid compost: anaerobic digestion, anaerobic polishing, nitrification, pH dosing, and / or filtration.

[0013] In an anaerobic step, pure cultures of carbon decomposers is cultivated to break down complex organic materials. In some embodiments, digestate from the anaerobic step may undergo an aerobic polishing step whereby aerobic heterotrophs are introduced to consume free organic acids, facilitating the survival and functionality of nitrifier digesters in subsequent steps. In some embodiments, a nitrification step may include reacting inorganic ions with oxygen, mediated by chemoautotrophs, to efficiently extract materials useful for plant nutrition and result in the conversion of ammonium into nitrate.

[0014] An aspect of the disclosure is directed to systems and methods for reducing high Biological Oxygen Demand (BOD) in irrigation water, which can lead to significant system fouling, the creation of anaerobic zones, and operational failures. BOD can be used as a measure of the amount of oxygen that microorganisms use to break down organic material present in water over a certain period, such as, for example, five days (BOD5) at 20°C. In some embodiments, the systems and methods may include one or more steps of the disclosed multi- step approach. In some embodiments, pre-digesting the feed reduces BOD and may convert nitrogen into forms such as ammonium, nitrite, and nitrate. In some embodiments, the use of nano or microfiltration on the digestate removes bacterial cells, sanitizes the solution, and eliminates suspended solids, while retaining large organic molecules in the retentate. In some embodiments, an appropriate selection of nano or microfiltration membranes may allow the passage of desired inorganic ions and organic molecules while blocking unwanted materials. Accordingly, the systems and methods of the disclosure may produce a biofertilizer that doesnot have high BOD or suspended solids. In some embodiments, a high level of BOD that is avoided would include a high BOD level, ranging from 20 mg / L to hundreds of mg / L, which indicates a high level of organic pollution. Accordingly, in some embodiments, a biofertilizer may have a BOD ranging from 0-7 mg / L, which can range from water with virtually no organic pollution to moderate levels of BOD that could lead to deteriorating water conditions as described herein. In some embodiments, the BOD is >1-2 mg / L. In some embodiments, the BOD is <2-3 mg / L. In some embodiments, the BOD is <3-4 mg / L. In some embodiments, the BOD is <4-5 mg / . In some embodiments, the BOD is <5-6 mg / L. In some embodiments, the BOD is <6-7 mg / L.

[0015] Such a biofertilizer may also be compatible with other processing techniques not available to traditional digestion effluents. Also, by utilizing nanofiltration to remove suspended solids, the effluent can then be directly dewatered with membrane technologies . For example, some embodiments may use Reverse Osmosis (RO) and / or Forward Osmosis (FO) may be used to dewater the effluent. RO systems can be used for removing a wide range of contaminants, including bacteria, viruses, salts, and organic compounds.

[0016] In some embodiments, a pH dosing step may actively adjust the pH levels to maintain a desirable environment for the digestion process(es). In some embodiments, the pH dosing is accomplished by one or more outputs of a reactor according to the disclosure. In some embodiments, the pH dosing step may convert the majority of nitrogen species into nitrate forms and allows for the stable coexistence of ammonia-oxidizing bacteria and nitriteoxidizing bacteria.

[0017] In some embodiments, a filtration step may involve the removal of suspended solids, tannins, and larger organic molecules from the water through nanofiltration and ultrafiltration, resulting in a solution predominantly composed of soluble salts. Accordingly, systems and methods of the disclosure allow for the production of fertilizers with useful plant nutrition and is compatible with irrigation systems. Preferred alternatives are provided below.

[0018] 1. A method of producing an organic fertilizer from a wastewater, comprising: introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent; wherein the anaerobic decomposition vessel comprises anorganic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained in an anaerobic condition at a pH from 6.0 to 9.0, transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel, digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent, wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea, transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel, digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or from 7.1 to 7.8; wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira,' and filtering at least the first or second aerobic effluent or both, wherein the second aerobic effluent comprises inorganic nitrogen in oxyanion form and has a carbon to nitrogen ratio greater than or equal to 3:1. Additional advantageous ranges of ORP are disclosed herein.

[0019] 2. The method of alternative 1, wherein at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from an inoculum.

[0020] 3. The method of alternative 2, wherein the anaerobic decomposition vessel comprises an inoculum selected from a group consisting of compost, worm castings, crop waste, microbial inoculum, and commercial inoculum, wherein the inoculum comprises a composition of microorganisms capable of digesting the organic material into at least organic acids and ammonium (but also include methane in here as additional scope) under basic conditions, said composition of microorganisms comprising bacteria known for their ability todecompose complex organic materials into simpler molecules, including organic acids, ammonium, and methane, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, Azotobacter, and Methanogens.

[0021] 4. The method according to any one of the preceding alternatives, wherein the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Escherichia coli, Escherichia fergusonii, Escherichia albertii, and Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae, and Methanogens including species such as Methanobacterium, Methanosarcina, Methanococcus, Methanomicrobium, Methanospirillum, Methanothermobacter, and Methanoculleus .

[0022] 5. The method according to any one of the preceding alternatives, wherein the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon decomposing bacteria, and wherein the plurality of microorganisms does not include ammonium decomposing bacteria.

[0023] 6. The method according to any one of the preceding alternatives, wherein the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of Lactobacillus, Streptococcus, and Enterobacter.

[0024] 7. The method according to any one of the preceding alternatives, wherein the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, e.g., species from the genera Bacteroides, Clostridium, or Eubacterium.

[0025] 8. The method according to any one of the preceding alternatives, wherein the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts compatible with a desired low oxygen and nonnitrifying environment for a least one of reduction of chemical and biological oxygen demand and consumption of organic acids.

[0026] 9. The method according to any one of the preceding alternatives, wherein the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentrationnot exceeding 2 mg / L (2 ppm), and further comprises a bacterial composition adapted to thrive in such low oxygen conditions.

[0027] 10. The method according to any one of the preceding alternatives, wherein the second aerobic decomposition vessel comprises an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria.

[0028] 11. The method according to any one of the preceding alternatives, wherein the at least one chemoautotrophic microorganism digests an ammonia compound present in the first aerobic effluent to produce nitrate compounds.

[0029] 12. The method according to any one of the preceding alternatives, wherein the second aerobic decomposition vessel comprises an environment facilitating decomposition of ammonia present in the first aerobic effluent and facilitating decomposition of nitrite compounds produced during decomposition of the ammonia.

[0030] 13. The method according to any one of the preceding alternatives, comprising maintaining the pH of a media in the second aerobic decomposition vessel at approximately 5.8, preferably by monitoring the pH in the second aerobic decomposition vessel and introducing the first aerobic effluent, which is preferably basic, from the first aerobic decomposition vessel into the second aerobic decomposition vessel when the pH falls below a pH of 5.8.

[0031] 14. The method according to any one of the preceding alternatives, wherein the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganisms and a plurality of nitrite oxidizer organisms.

[0032] 15. The method according to any one of the preceding alternatives, further comprising raising the pH of a media in the second aerobic decomposition vessel by transferring the first aerobic effluent from the first aerobic decomposition vessel into the second aerobic decomposition vessel.

[0033] 16. The method of alternative 15, wherein an amount of the first aerobic effluent is added to the second aerobic decomposition vessel sufficient to raise the pH of the media in the second aerobic decomposition vessel to 6.2 or 7.8.

[0034] 17. The method according to any one of the preceding alternatives, wherein a process of digesting the first aerobic effluent in the second aerobic decomposition vessel decreases (acidifies) the pH of a media in the second aerobic decomposition vessel.

[0035] 18. The method according to any one of the preceding alternatives, further comprising adjusting the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 or 7.1 to 7.8 (or 7.8 to 7.1) by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0036] 19. The method of alternative 18, wherein the amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel is transferred when the pH of the media in the second aerobic decomposition vessel reaches 5.5 until the pH of the media in the second aerobic decomposition vessel reaches 6.2.

[0037] 20. The method according to any one of the preceding alternatives, wherein transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel is performed multiple times so as to provide a continuous culture where a majority of any nitrogen species are in nitrate form; and wherein a population of ammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions. The addition of ozone or an oxidizer treatment to the membrane permeate may also serve to further convert nitrite to nitrate. This process is useful for maintaining the stability and shelf life of the final product. During fermentation, nitrites can accumulate, which are less stable and potentially harmful. By introducing ozone or an oxidizer, these nitrites are oxidized to nitrates, which are more stable and less reactive. This conversion not only ensures a more stable product but also enhances the overall safety and quality of the fermentation output. This additional treatment step effectively stabilizes the permeate, transforming it into a more suitable form for long-term storage and use.

[0038] 21. The method according to any one of the preceding alternatives, wherein the filtration further comprises a prefiltration process using a large pore microfilter to remove particulate matter, preferably followed by a tubular nanofilter, and preferably subsequently followed a spiral ultrafilter. As described herein, ultrafiltration may be useful due to its efficiency in removing fine particulates and contaminants, but other membrane filtration techniques can also be sufficient if they achieve similar reductions in suspended solids and BOD.

[0039] 22. The method according to alternative 21, wherein the sequence of filtration removes suspended solids, tannins, and larger organic molecules from the second aerobic effluent.

[0040] 23. The method according to alternatives 21 or 22, wherein a retentate from the ultrafiltration process is returned to at least one of the first aerobic decomposition vessel and the second aerobic decomposition vessel.

[0041] 24. The method according to any one of the preceding alternatives, further comprising a side stream process, wherein a portion of a filtered effluent is recycled during the filtration.

[0042] 25. The method according to any one of the preceding alternatives, wherein a discharge of a filtered effluent is performed, wherein said discharge removes a portion of the accumulated suspended solids.

[0043] 26. The method according to any one of the preceding alternatives, wherein the filtration comprises a sequential filtration process, first employing an ultrafiltration and then employing a nanofiltration, preferably in a series, wherein the nanofiltration follows the ultrafiltration.

[0044] 27. The method according to any one of the preceding alternatives, wherein, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3:1.

[0045] 28. The method according to any one of the preceding alternatives, wherein the anaerobic effluent comprises carbon decomposition products, such as organic acids.

[0046] 29. The method according to any one of the preceding alternatives, wherein the first aerobic effluent has a reduced chemical oxygen demand as compared to the anaerobic effluent.

[0047] 30. The method according to any one of the preceding alternatives, wherein the first aerobic effluent is substantially free of oxyanions of nitrogen.

[0048] 31. The method according to any one of the preceding alternatives, wherein the second aerobic effluent comprises nitrates.

[0049] 32. The method according to any one of the preceding alternatives, wherein the second aerobic effluent comprises reduced amounts of ammonia compared to the first aerobic effluent.

[0050] 33. The method according to any one of the preceding alternatives, further comprising adjusting the pH of the anaerobic effluent, the first aerobic effluent, and / or the second aerobic effluent.

[0051] 34. The method according to any one of the preceding alternatives, further comprising adding at least one of base and a carbon source, such as molasses, saccharides, lime, and bicarbonates, to the second aerobic decomposition vessel and / or the first aerobic decomposition vessel.

[0052] 35. The method according to any one of the preceding alternatives, wherein the ORP is measured using an electrochemical sensor comprising a platinum measurement electrode and a silver / silver chloride reference electrode.

[0053] 36. The method according to any one of the preceding alternatives, wherein the ORP is measured using a colorimetric testing module configured to use redox indicators for estimating the ORP of a media in a digestion vessel.

[0054] 37. The method according to any one of the preceding alternatives, further comprising adding at least one of a peroxide, ozone, humic acid and / or fulvic acid to the second aerobic effluent.

[0055] 38. A system for producing an organic fertilizer from wastewater, comprising: at least one anaerobic digestion vessel configured to receive an organic feed stock, such as a wastewater comprising an organic waste, wherein the at least one anaerobic digestion vessel provides an anaerobic environment suitable to produce an anaerobic effluent, wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0, at least one aerobic oxidation vessel in fluid communication with the at least one anaerobic digestion vessel, wherein the at least one aerobic oxidation vessel is configured to receive anaerobic effluent from the at least one anaerobic digestion vessel and oxygen, and digest the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent, wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea;and at least one aerobic nitrification vessel in fluid communication with the at least one aerobic oxidation vessel, wherein the at least one aerobic oxidation vessel is configured to receive the first aerobic effluent and oxygen, and to digest the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8, wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira.

[0056] 39. The system according alternative 38, further comprising a port configured to receive an acid, a base or a carbon source, wherein said port is connected to at least one of the anaerobic digestion vessel, the aerobic oxidation vessel, and / or the aerobic nitrification vessel.

[0057] 40. The system according to any one of alternatives 38 to 39, wherein at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from an inoculum, preferably an inoculum of worm castings and / or crop waste.

[0058] 41. The system according to any one of alternatives 38 to 40, wherein the anaerobic decomposition vessel comprises an inoculum comprising a composition of microorganisms capable of digesting the organic material into organic acids under basic conditions, said composition of microorganisms comprising bacteria known for their ability to decompose complex organic materials into simpler molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter.

[0059] 42. The system according to any one of alternatives 38 to 41, wherein the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens Escherichia coli, Escherichia fergusonii, Escherichia albertii; Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae, and Methanogens includiing species such as Methanobacterium, Methanosarcina, Methanococcus, Methanomicrobium, Methanospirillum, Methanothermobacter, and Methanoculleus .

[0060] 43. The system according to any one of alternatives 38 to 42, wherein the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon decomposing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria.

[0061] 44. The system according to any one of alternatives 38 to 43, wherein the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter.

[0062] 45. The system according to any one of alternatives 38 to 44, wherein the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, and Eubacterium.

[0063] 46. The system according to any one of alternatives 38 to 45, wherein the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts compatible with a desired low oxygen and nonnitrifying environment for at least one of reduction of chemical and biological oxygen demand and of consumption of organic acids.

[0064] 47. The system according to any one of alternatives 38 to 46, wherein the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), and further comprises a bacterial composition adapted to thrive in such low oxygen conditions.

[0065] 48. The system according to any one of alternatives 38 to 47, wherein the second aerobic decomposition vessel comprises an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria.

[0066] 49. The system according to any one of alternatives 38 to 48, wherein the at least one chemoautotrophic microorganism digests an ammonia compound present in the first aerobic effluent to produce nitrate compounds.

[0067] 50. The system according to any one of alternatives 38 to 49, wherein the second aerobic decomposition vessel comprises an environment facilitating decomposition ofammonia present in the first aerobic effluent and facilitating decomposition of nitrite compounds produced during decomposition of the ammonia.

[0068] 51. The system according to any one of alternatives 38 to 50, wherein the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganism and a plurality of nitrite oxidizer organism.

[0069] 52. The system according to any one of alternatives 38 to 51, wherein the pH of a media in the second aerobic decomposition vessel is raised by transferring the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0070] 53. The system according to any one of alternatives 38 to 52, wherein the second aerobic decomposition vessel is configured to digest the first aerobic effluent in the second aerobic decomposition vessel and increase the pH of a media in the second aerobic decomposition vessel.

[0071] 54. The system according to any one of alternatives 38 to 53, wherein the second aerobic decomposition vessel is configured to adjust the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 or 7.1 to 7.8 by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0072] 55. The system according to any one of alternatives 38 to 54, wherein the second aerobic decomposition vessel is configured to transfer an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel when the pH of the media in the second aerobic decomposition vessel reaches 5.5, until the pH of the media in the second aerobic decomposition vessel reaches 6.2.

[0073] 56. The system according to any one of alternatives 38 to 55, wherein the second aerobic decomposition vessel is configured to transfer the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel multiple times so as to provide a continuous culture where a majority of any nitrogen species are in nitrate form; and wherein a population of ammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions.

[0074] 57. The system according to any one of alternatives 38 to 56, further comprising a prefiltration unit, a microfilter with a pore size between 1 and 10 micrometers to remove particulate matter, a spiral ultrafilter, and a tubular nanofilter. Typically only one of a nanofilter or an ultrafilter may be utilized, but in some cases they might be operated in series. Typically, a microfilter may precede an ultra or nano, as described.

[0075] 58. The system according to any one of alternatives 38 to 57, wherein the filter is configured to remove suspended solids, tannins, and larger organic molecules from the second aerobic effluent.

[0076] 59. The system according to any one of alternatives 38 to 58, wherein a retentate from the ultrafiltration process is returned to at least one of the first aerobic decomposition vessel or the second aerobic decomposition vessel.

[0077] 60. The system according to any one of alternatives 38 to 59, further comprising a side stream unit configured to recycle a portion of the retentate during the filtration step.

[0078] 61. The system according to any one of alternatives 38 to 60, wherein, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1. In some embodiments, the same effluent may reside in a storage tank under aeration. In some embodiments, extra nitrification can occur here in the event a stage 3 is substantially nitrite.

[0079] 62. The system according to any one of alternatives 38 to 61, wherein the anaerobic effluent comprises carbon decomposition products, such as organic acids.

[0080] 63. The system according to any one of alternatives 38 to 62, wherein the first aerobic effluent has a reduced chemical oxygen demand as compared to the anaerobic effluent.

[0081] 64. The system according to any one of alternatives 38 to 63, wherein the first aerobic effluent is substantially free of oxyanions of nitrogen.

[0082] 65. The system according to any one of alternatives 38 to 64, wherein the second aerobic effluent comprises nitrites.

[0083] 66. The system according to any one of alternatives 38 to 65, wherein the second aerobic effluent comprises reduced amounts of ammonia compared to the first aerobic effluent.

[0084] 67. The system according to any one of alternatives 38 to 66, wherein theORP is measured using an electrochemical sensor comprising a platinum measurement electrode and a silver / silver chloride reference electrode.

[0085] 68. The system according to any one of alternatives 38 to 67, wherein theORP is measured using a colorimetric testing module configured to use redox indicators for estimating the ORP of a media in a digestion vessel.

[0086] 69. A method of producing an ammonium-based biofertilizer from a wastewater, including introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent, wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0, transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel; and digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea.

[0087] 70. The method of alternative 69, further comprising filtering and dewatering the first aerobic effluent and producing a concentrated filtrate.

[0088] 71. The method according to any one of alternatives 69 to 70, wherein the anaerobic decomposition process is at least one of substantially acetogenic, substantially hydrolytic, and substantially methanogenic.

[0089] 72. The method according to any of one of alternatives 69 to 71, wherein at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from an inoculum comprising at least one of worm castings, crop waste, microbial inoculum, and commercial inoculum.

[0090] 73. The method according to any of alternatives 69 to 72, wherein the anaerobic decomposition vessel comprises a composition of microorganisms capable ofdigesting the organic material into organic acids under basic conditions, said composition of microorganisms comprising bacteria known for their ability to decompose complex organic materials into simpler molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter, and Methanogens such as Methanobacterium, Methanosarcina, Methanococcus, Methanomicrobium, Methanospirillum, Methanothermobacter, and Methanoculleus .

[0091] 74. The method according to any one of alternatives 69 to 73, wherein the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Escherichia coli, Escherichia fergusonii, and Escherichia albertii, Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae.

[0092] 75. The method according to any one of alternatives 69 to 74, wherein the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon decomposing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria.

[0093] 76. The method according to any one of alternatives 69 to 75, wherein the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter.

[0094] 77. The method according to any one of alternatives 69 to 76, wherein the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, and Eubacterium.

[0095] 78. The method according to any one of alternatives 69 to 77, wherein the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts sufficient to maintain the desired low oxygen and non-nitrifying environment for the consumption of organic acids.

[0096] 79. The method according to any one of alternatives 69 to 78, wherein the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration notexceeding 2 mg / L (2 ppm), and further comprises a bacterial composition adapted to thrive in such low oxygen conditions.

[0097] 80. The method according to any one of alternatives 69 to 79, wherein the method is operated continuously, and further wherein the hydraulic retention time of the solution acidified with an acid in the oxidation reactor is sufficient to substantially decompose the acid, such as substantially decomposing the base component of acetic acid — acetate. In some embodiments, a commercially available citric acid or acetic acid may be introduced directly into the first aerobic vessel for pH adjustment. In this process the first aerobic digestor may be pH corrected with, for example, citric acid to lower the pH and remove bicarbonate, whereby the base component of the citric acid would then be digested out.

[0098] 81. The method according to any one of alternatives 69 to 79, wherein the method is operated in batches, and wherein the hydraulic retention time of the solution acidified with the acid in the oxidation reactor is sufficient to substantially decompose the base component of the acid, such as substantially decomposing the base component of acetic acid — acetate.

[0099] 82. The method according to any one of alternatives 69 to 81, wherein the biofertilizer is compatible as a component of YAN (Yeast Assimilable Nitrogen).

[0100] 83. The method according to any one of alternatives 69 to 82, further comprising the addition of an oxidizer such as Ozone or hydrogen peroxide to further oxidize any ammonium or nitrite to nitrate in a filtrate. Ozone may be used as a sanitizer and can fully oxidize ammonia and nitrite to nitrate. So, the ozone can be used for sanitation with the inherent benefit of nitrogen conversion. This method works best on direct membrane filtrate or an irrigation water where the COD is low.

[0101] 84. A liquid fertilizer produced by an organic waste digester, wherein the liquid fertilizer has a biological oxygen demand (BOD) of less than 100 mg / L.

[0102] 85. The liquid fertilizer of alternative 84, wherein the liquid fertilizer is stable upon addition of peroxides. In some embodiments, stability may be determined when the COD remains high after addition and peroxide is stable in solution and does not react with compounds providing the COD.

[0103] 86. The liquid fertilizer of alternative 84 or 85, further comprising an organic acid.

[0104] 87. The liquid fertilizer of any one of alternatives 84 to 86, further comprising a carbonaceous species to raise the C:N ratio, such as an organic acid selected from the group consisting of humic acid, fulvic acid, citric acid, and acetic acid.

[0105] 87. The liquid fertilizer of any one of alternatives 84 to 87, wherein the liquid fertilizer is further treated with the addition of an oxidizer such as Ozone or hydrogen peroxide to further oxidize any ammonium or nitrite to nitrate.

[0106] 88. The liquid fertilizer of any one of alternatives 84 to 87, wherein the composition is produced by introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent; wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0, transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel, digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea, transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel; and digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira, e.g., a bacterial population obtained from earthworm castings.

[0107] 89. A method according to any of the preceding alternatives, further including in certain embodiments, at least one base media contactor, which is fluidicallyconnected to at least one aerobic oxidation vessel, such as the second aerobic decomposition vessel, and functions to modulate solution pH and supplement mineral ions that are beneficial to microbial activity and biofertilizer quality. The base media contactor comprises a bed of alkaline-reactive solids selected from calcite, dolomitic limestone, phosphate rock, magnesium oxide, Corosex™, or combinations thereof. The contactor may accommodate either upflow or downflow fluid dynamics. In an upflow arrangement, the residence time of the fluid within the contactor is controlled to enable dissolution of alkaline constituents, particularly carbonates and oxides, thereby elevating the pH of the process stream. In a downflow arrangement, the contactor may additionally serve as a filtration medium, mechanically retaining suspended solids and particulates before exposure to downstream filtration stages such as microfiltration, ultrafiltration, or nanofiltration units.

[0108] 90. A method according to any of the preceding alternatives, further including in certain embodiments, the base media contactor is operated in closed-loop feedback with real-time monitoring of parameters within the third aerobic decomposition vessel. When sensor inputs indicate elevated nitrite levels or declining bicarbonate availability in Stage 3, the operational throughput or contact duration within the base media contactor is adjusted to increase the dissolution rate of alkaline solids. This responsive operation supports microbial community stability and mitigates nitrite inhibition, particularly within nitrifying populations belonging to genera such as Nitrosomonas, Nitrobacter, and Nitrospira.

[0109] 91. A method according to any of the preceding alternatives, further including in certain embodiments, the controlled dissolution of carbonate or oxide-based components within the base media contactor introduces both alkaline anions (e.g., carbonate, bicarbonate, hydroxide) and agronomically beneficial counterions (e.g., calcium, magnesium, phosphate). The availability of these ions supports enzymatic function and osmotic balance in microbial cultures, while also enhancing the nutritional value of the resulting biofertilizer product. The mineral composition of the base media is selected to comply with organic agricultural certification standards, thereby making the system compatible with organic market requirements.

[0110] 92. A method according to any of the preceding alternatives, further including in certain embodiments, the downflow operation of the base media contactor is specifically utilized for pre-filtration of effluent from Stage 2. This function reduces theloading of suspended solids and particulate matter prior to exposure to microfilters, spiral ultrafilters, or tubular nanofilters. By reducing fouling potential, this pre-filtration step contributes to the maintenance of stable transmembrane pressures and extends the operational lifespan of downstream filtration membranes.

[0111] 93. A method according to any of the preceding alternatives, further including in certain embodiments, the dissolution of mineral constituents in the base media contactor is coordinated with broader pH management strategies across the multi-stage decomposition system. For instance, mineral ion release may be synchronized with the codosing of aqueous ammonia during Stage 2 operations, such that ammonia-nitrogen concentrations and pH levels are concurrently optimized for downstream nitrification. This integrated chemical management approach improves nitrification rates in Stage 3 and supports a continuous, rather than batch-mode, operation, distinguishing the system from conventional treatment designs.

[0112] 94. A method according to any of the preceding alternatives, further including in certain embodiments, Stage 3 operation is supplemented by exposure to the effluent modified by the base media contactor when pH cycling strategies are employed. When inhibitory nitrite concentrations are detected, pH may be transiently lowered followed by a rapid recovery using mineral-enriched alkaline solutions. This pH cycling supports microbial recovery and helps maintain consistent biofertilizer output quality, particularly when integrated with upstream aeration and retention time adjustments in Stage 2.BRIEF DESCRIPTION OF THE DRAWINGS

[0113] Fig. 1 is a flowchart of a nonlimiting example method for producing an organic biofertilizer from a wastewater.

[0114] Fig. 2 illustrates a multi-stage system for converting a raw organic feed to a filtered output, such as a biofertilizer or component thereof, suitable for use in an irrigation system.

[0115] Fig. 3 is a graphical representation of experimental pH levels in a reaction over time, where the pH cycles in a wave-like pattern.

[0116] Fig. 4 is a pie chart summarizing the percentages by class of microorganisms in a stage 1 tank.

[0117] Fig. 5 is a pie chart summarizing the percentages by class of microorganisms in a stage 2 tank.

[0118] Fig. 6 is a pie chart summarizing the percentages by class of microorganisms in a stage 3 tank.

[0119] Fig. 7 shows a schematic diagram of a reactor system for converting a raw organic feed input into a biofertilizer or component thereof showing different stages indicated by pipes, which illustrate a sequential process flow.DETAILED DESCRIPTION

[0120] The present disclosure provides systems and methods for producing a biofertilizer or a biofertilizer component from agricultural waste and / or wastewater, including for example, introducing the wastewater into one or more decomposition vessels configured to produce effluents under anaerobic or aerobic conditions. Despite several advances in bioreactor designs and biofertilizer production processes, conventional processes and bioreactors fail to employ the presently described systems and methods e.g., utilization of the specific bacterium types employed at the various decomposition stages utilized, providing an aerobic polishing step or providing nitrification steps, which convert ammonium salt into nitrate. By incorporating these unique elements into the processes described herein, improved biofertilizers and components thereof are obtained.

[0121] While there exist publications that discuss the feasibility of certain aspects of bioreactor designs, they fall short of teaching or suggesting the specific elements found to unexpectedly improve the production of biofertilizers and components thereof. Chinese Patent No. 108610106, expressly incorporated by reference in its entirety, outlines a method for producing humic acid fertilizer from agricultural wastes and methane fermentation. U.S. Publication No. 2009 / 0282882, expressly incorporated by reference in its entirety, outlines a method for treating waste biomass to produce a fertilizer. U.S. Patent No. 9,716,038, expressly incorporated by reference in its entirety, outlines an ammonia recovery system that extracts ammonia gas to form an acidic ammonium salt solution, with the possibility of further acidification. In contrast to these teachings, the present disclosure provides an advanced method for transforming agricultural waste and / or wastewater into a nutrient-rich organic fertilizer, optimized for effective use in agricultural applications.

[0122] Some embodiments relate to one, two, three or more stage bioreactors for digesting biomaterials. In some embodiments, specific cultures, strains and / or species of microorganisms are described as being contained in one or more bioreactors. In some embodiments, the disclosure provides for methods of inoculating, culturing, or otherwise encouraging growth or death of such microorganisms. Some embodiments relate to the types of effluents generated by one or more bioreactors. In some embodiments, the effluent of one or more bioreactors is used as a source of fertilizer or a source of methane. Various embodiments describe the production or different types of fertilizers depending on the method of operation of the bioreactors. As described herein, it was surprisingly discovered that typical effluent streams could be adapted into producing fertilizers that are compatible with irrigation systems e.g., reduce clogging of equipment, especially nozzles and sprinklers, which deploy the fertilizer. Additionally, certain biofertilizers release nutrition from biomaterials and, also prevent odors and the buildup of detritus inside an irrigation system.

[0123] One goal of wastewater treatment is the complete decomposition and mineralization of complex waste waters, followed by nutrition removal. In other wastewater processes, a mineralization step occurs first, then nitrogen and phosphorus are removed. The goal in these approaches is to break down the waste as much as possible, then remove nutrition so water is not polluted with nitrogen and phosphorus once it is discharged back to the environment. Feeding microorganisms too much readily available nutrition can cause environmental disasters, such as algae blooms. The instant disclosure provides methods to mineralize the biomaterials, but not proceed down a path that would be used to remove nitrogen and phosphorus so as to retain the beneficial, readily available nutrition.

[0124] Before the present disclosure is described further, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0125] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the disclosure. The upper and lower limits of thesesmaller ranges may independently be included in the smaller ranges and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0126] Methods recited herein may be carried out in any order of the recited events, which is logically possible, as well as the recited order of events.

[0127] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described.

[0128] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0129] It must be noted that as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.Definitions

[0130] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. In the event that there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0131] Digestate: A byproduct or output of a digestion process, where typically organic materials are broken down by microorganisms in the absence of oxygen. This process is commonly used for the treatment of waste materials, such as sewage sludge, agricultural waste, and organic industrial waste. Digestate may include indigestible material and deadmicroorganisms, and may be rich in nutrients such as nitrogen, phosphorus, and potassium that may or may not be accessible to plants as nutrients. As such, it is often used as a soil conditioner or fertilizer, helping to improve soil health and support plant growth.

[0132] Biochemical Oxygen Demand (BOD): A measure of the amount of oxygen that microorganisms use to decompose organic matter in water. BOD is a parameter in water quality assessment and may be used to gauge the effectiveness of wastewater treatment processes.

[0133] Bioreactor: A device or system that supports a biologically active environment. This term may be used interchangeably with reactor, or vessel. Bioreactors are typically used for growing cells, fermenting microorganisms, or conducting enzymatic reactions. Different vessels may be designed to provide the optimal conditions for the desired biological process, including temperature, pH, oxygen levels, and nutrient supply. Bioreactors can range in size from small laboratory-scale units to large industrial-scale systems, depending on their use.

[0134] Mineralization: The process of converting organic matter into minerals or inorganic compounds, often through bacterial or enzymatic action.

[0135] Organic feeds: Substances derived from living organisms, such as animal or plant waste, which are used as a source of nutrients in various industrial and agricultural processes.

[0136] Activated sludge: A mixture of wastewater and microorganisms, such as bacteria and protozoa, which are used to break down organic matter in sewage and industrial wastewater.

[0137] Aerobic bacteria: Microorganisms that require oxygen to grow and reproduce.

[0138] Digestor: A vessel or tank used to break down organic matter through bacterial or enzymatic action.

[0139] Aeration: The process of adding oxygen to a substance, such as water or soil, to support the growth of aerobic bacteria or other organisms.

[0140] Municipal waste: Solid or liquid waste generated by households, businesses, or other institutions in a city or town.

[0141] Heterotrophic bacteria: Microorganisms that obtain energy and carbon from organic matter, such as sugars or proteins, produced by other organisms.

[0142] Autotrophic nitrifiers: Bacteria that obtain energy from inorganic compounds and convert ammonia into nitrite and nitrate, which can be used by other organisms as a source of nitrogen.

[0143] Denitrification: The process by which nitrate and nitrite are converted into nitrogen gas, which can be released into the atmosphere.

[0144] Carbon-to-carbon bonds: Chemical bonds that connect carbon atoms in a molecule, which are often rich in energy and can be broken down by bacteria to obtain energy and carbon for their own cells.

[0145] CO2: Carbon dioxide, a gas produced by heterotrophic bacteria as a byproduct of respiration.

[0146] Aerobic: Referring to a process or organism that uses oxygen to grow and reproduce.

[0147] Microaerophilic: Referring to a condition wherein a small amount of oxygen is present, which some bacteria, including heterotrophs, can survive in.

[0148] Growth rate: The rate at which a population of bacteria increases in number over time.

[0149] Carbonaceous feed: Organic matter that serves as a source of carbon for heterotrophic bacteria.

[0150] Nutrients: Substances that are essential for the growth and survival of bacteria, including elements such as nitrogen, phosphorus, and sulfur, and vitamins and other organic compounds.

[0151] Logarithmic growth: A rapid phase of bacterial growth characterized by exponential increase in numbers of individuals.

[0152] Macromolecules: Large molecules made up of many smaller subunits, such as proteins, starches, and fats.

[0153] Enzymes: Proteins that catalyze chemical reactions, including the breakdown of large molecules into smaller ones that can be absorbed by bacteria.

[0154] Monomers: Smaller subunits that make up larger molecules, which can be absorbed by bacteria for use as nutrients.

[0155] Hydraulic retention time: The length of time that wastewater remains in a treatment system, which can affect the rate of decomposition and mineralization by heterotrophic bacteria.

[0156] Wastewater: Wastewater includes any water that includes organic materials. Common examples of wastewater include sewage from households, runoff from agricultural fields carrying fertilizers, and discharge from industrial processes. Wastewater as referred to herein can also include solid waste materials, such as biomaterials, which are added to a vessel or container, leading to the formation of a mixture with water. These solid wastes may include organic matter, such as food scraps or plant residues, which when combined with water can create a slurry or semi-liquid waste.

[0157] Agricultural waste: Agricultural waste refers to any by-products or residues generated from agricultural activities. This includes a wide range of materials such as crop residues (stalks, leaves, husks, and shells), livestock manure, poultry litter, and other organic wastes produced from agricultural operations. These wastes can originate from various agricultural practices, including crop cultivation, animal husbandry, aquaculture, and agroindustrial processes. While often considered a disposal challenge, agricultural waste also presents opportunities for recycling and reuse in sustainable farming practices, such as composting, bioenergy production, and as raw materials for bio-based products.

[0158] Biofertilizer: Biofertilizers refer to substances that contain living microorganisms or residues thereof, which, when applied to seeds, plant surfaces, or soil, may promote growth by increasing the supply of primary nutrients to a host plant.

[0159] Anaerobic: In the context of low oxygen environments, "low" signifies a gas or liquid concentration of oxygen that is below those typically found in standard atmospheric conditions. By way of non-limiting examples, a range of oxygen levels consistent with a low oxygen level might encompass concentrations below 200,000 parts per million (ppm). Low oxygen levels include oxygen levels of or approximately of 1,000 to 200,000 ppm, to near-zero oxygen levels, where concentrations approach 0 ppm, indicative of anoxic conditions. Low oxygen levels may also independently refer to oxygen levels of or approximately of 10,000 to 200,000 ppm, 20,000 to 190,000 ppm, 30,000 to 180,000 ppm, 40,000 to 170,000 ppm, 50,000 to 160,000 ppm, 60,000 to 150,000 ppm, 70,000 to 140,000 ppm, 80,000 to 130,000 ppm, or 90,000 to 120,000 ppm. Low oxygen levels may alsoindependently refer to oxygen levels of or approximately of 1 to 200 ppm, 2 to 19 ppm, 3 to 18 ppm, 4 to 17 ppm, 5 to 16 ppm, 6 to 15 ppm, 7 to 14 ppm, 8 to 13 ppm, or 9 to 12 ppm. For the purposes of anaerobic vessels, the vessel may, for example, not have a low oxygen environment immediately after the addition of an input. Low oxygen levels may also independently refer to oxygen levels of less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2, and less than 1 ppm. Low oxygen levels may also independently refer to oxygen levels that are substantially anoxic. In various embodiments, the anaerobic may refer to one or both of the concentration of oxygen in the gas phase and the concentration of oxygen in the liquid phase. Typical levels of oxygen in the gas phase at atmospheric conditions is 200,000 ppm. Typical levels of oxygen of oxygen in the liquid phase, also referred to as dissolved oxygen, is 5-10 ppm.

[0160] Chemoautotrophic growth refers to growth of an organism that depends on its energy gain via a non-organic (non-carbon based) Oxidation or Reduction (chemo-) and its carbon source is from CO2 (solubilized in water as bicarbonate / carbonate).Fertilizer Production Methods

[0161] An aspect of this disclosure relates to methods of fertilizing a crop or treating a wastewater or agricultural waste using an organic fertilizer supply. Microorganisms can decompose most natural and many synthetic products into salts and stable humic material, but entail growing and sustaining a large and diverse group of organisms for this decomposition to occur. Organic feeds are very diverse, but for the most part, they typically are a minimally refined waste products from food production industries. Taking these complex feeds and utilizing them in performance indoor cropping systems using complex plumbing systems presents many challenges not typical of traditional conventional crop production. By contrast, a typical indoor crop system might use conventional fertilizer and maintain the ideal nutrient solution by automatically injecting a high concentration of nutrient salts into water, monitoring the electrical conductivity (EC) of the solution to maintain a consistent level of nutrients by adding more of the salt mixture if the EC drops due to addition of water or plant uptake. Organic fertilizers, on the other hand, often have salts bound up in large organic molecules, which are released after microbial action to decompose them into ions. Microbial cultures can also depend on the carbon content of the feed and their waste products are the saltleftover, which is the nutritional salts the plant can use. Accordingly, using organic fertilizer in, for example, an irrigation system may not be as simple as switching from conventional fertilizers.

[0162] Systems and methods according to the disclosure are able to utilize organic fertilizers and extract a whole range of nutrients that in many cases are sufficient for a general- purpose fertilizer. In some cases, the disclosure also provides for methods of producing a fertilizer and provides for novel and useful fertilizers. Additionally, the disclosure provides for systems compatible with hosting microbial decomposition processes, such as the non-limiting examples of irrigation systems, greenhouses and aquaponics systems.

[0163] Fig. 1 is a flowchart of a nonlimiting example method for producing an organic fertilizer from a wastewater. The method may begin at step 110, by introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent. In some embodiments, the anaerobic decomposition vessel may include an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose. In some embodiments, the organic material is preferably a food or crop material. In some embodiments, the anaerobic decomposition vessel is maintained in an anaerobic condition at a pH from 6.0 to 9.0.

[0164] After sufficient decomposition, which is preferably monitored by methods according to the disclosure, the anaerobic effluent is transferred at step 120, from the anaerobic decomposition vessel to a first aerobic decomposition vessel. In some embodiments, the transfer occurs, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel.

[0165] Once transferred to the first aerobic decomposition vessel, the anaerobic effluent is digested at step 130, in the presence of oxygen or an oxidizer. In some embodiments, the oxidizer is hydrogen peroxide, or ozone. In some embodiments, the anaerobic effluent is digested while maintaining a pH from 7.0 to 9.0. In some embodiments, the anaerobic effluent is digested at an oxidation reduction potential (ORP) between -1000 mV and 1000 mV. In some embodiments, the ORP intervals may span from -1500 mV to 1500 mV, -1200 mV to 1200 mV, the original -1000 mV to 1000 mV, -800 mV to 800 mV, -500 mV to 500 mV, -700 mV to 700 mV, and finally, a highly specific -100 mV to 100 mV. In some embodiments, the ORP may range from -1000, -900, -800, -700, -600, -500, -700, -700, -200, -100, 0, 100, 200,700, 700, 500, 600, 700, 800, 900, to 1000 mV. In bioreactor operations, where the oxidationreduction potential (ORP) plays a role in driving specific biochemical reactions, several potential alternatives across different processes can be considered. For anaerobic digestion, maintaining an ORP between -700 mV and -200 mV is typical. Aerobic digestion processes, relevant for wastewater treatment more generally, typically employ a more positive ORP range of +100 mV to +700 mV to support aerobic microorganisms in consuming organic pollutants. In the realm of nitrogen removal, nitrification may use a slightly positive ORP of +200 mV to +700 mV, whereas denitrification, which converts nitrate may operate at an ORP from -200 mV to 0 mV. Additionally, for fermentation processes or mixed culture cultivation, the ORP can vary significantly, generally within -700 mV to +700 mV, depending on the target product and involved microorganisms. In some embodiments, the first aerobic vessel comprises at least one organic acid consuming organism. In some embodiments, an organic acid consuming organism is a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea. After digestion, the anaerobic effluent will produce a first aerobic effluent.

[0166] The first aerobic effluent is transferred at step 140 to another vessel. In some embodiments, the first aerobic effluent is transferred from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel. Once transferred to the second aerobic decomposition vessel, the first aerobic effluent is digested at step 150, in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer. In some embodiments, the oxidizer is hydrogen peroxide, or ozone.

[0167] In some embodiments, the first aerobic effluent is digested while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8. In some embodiments, the second aerobic decomposition vessel includes at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira.

[0168] In some embodiments, the first aerobic effluent is digested to produce a second aerobic effluent. The process may optionally conclude at step 160, by filtering at least the first or second aerobic effluent or both. In some embodiments, the preceding steps address the problem of maintaining sanitizer levels in irrigation water, which can be a significant challenge in other systems due to either difficulty or high costs. Some embodiments utilizenanofiltration or microfiltration, which can significantly reduce the content of organic molecules that absorb UV light, thus allowing for the effective use of UV light for sanitization. Furthermore, in some embodiments, reducing suspended solids in the system allows for using hydrogen peroxide as a sanitizer. In some embodiments, the hydrogen peroxide is a preferred oxidizer to ozone. In some embodiments, the second aerobic effluent includes inorganic nitrogen in an oxyanion form. In some embodiments, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1, such as 3: 1, 4:1, 5: 1, 6:1, 7: 1, or 8: 1 or more.

[0169] In some embodiments, the systems and methods according to the disclosure include introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent, transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, and digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer to produce a second aerobic effluent.

[0170] In some embodiments, the systems and methods according to the disclosure include introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent. The systems and methods discussed in the context of treating wastewater or agricultural waste through anaerobic decomposition may involve managing and processing the waste. These embodiments typically entail using a dedicated vessel designed for anaerobic decomposition, which functions under conditions devoid of oxygen. The primary input to this process is wastewater or liquified agricultural waste, such as agricultural waste suspended in water, which can vary widely in composition based on its source, including domestic, industrial, or agricultural waste or runoff. This wastewater usually contains organic matter, microorganisms, and may require treatment or sterilization before the water obtained after the process can be safely discharged into the environment or reused.

[0171] In some embodiments, the anaerobic decomposition vessel includes an organic material, such as an organic material including a sugar, protein, lipid or cellulose, preferably a food or crop material. Such materials are often derived from food waste oragricultural by-products, making them readily available and sustainable sources for anaerobic digestion processes. The choice of these materials may impact any nutrients in the resulting fertilizer and may also impact the rate at which the feed is input into the first stage.

[0172] In some embodiments, the anaerobic decomposition vessel is maintained in an anaerobic condition at a pH from 6.0 to 9.0. Without being bound to a single theory of operation, this pH range will help in stabilizing the microbial activity, ensuring that acidogenic bacteria and methanogens, for example, can thrive and efficiently break down the organic material into biogas.

[0173] The output of this anaerobic decomposition process is referred to as an anaerobic effluent. This effluent would include the treated or partially treated water and byproducts of the anaerobic digestion process, which may include a biogas — a mixture of methane, carbon dioxide, and traces of other gases. The state of the outputs may be significantly different from the inputs in that the organic load of the wastewater is substantially reduced, making the effluent less polluting and sometimes suitable for discharge into water bodies or further treatment. The process also reduces the volume of sludge produced compared to aerobic processes, offering a more sustainable and cost-effective solution for wastewater management. The biogas produced can be captured and used as a source of renewable energy, further enhancing the environmental benefits of this method.

[0174] In some embodiments, systems and methods according to the disclosure may include transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel. In some embodiments, the anaerobic effluent is transferred by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel. In some embodiments, systems and methods according to the disclosure may include digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone to produce a first aerobic effluent.

[0175] In some embodiments, the first aerobic decomposition vessel is maintained at a pH from 7.0 to 9.0, In some embodiments, the first aerobic decomposition vessel is maintained at an oxidation reduction potential (ORP) between -1000 mV and 1000 mV. The maintenance of specific pH and / or ORP (Oxidation-Reduction Potential) ranges in aerobic decomposition vessels is useful for optimizing the breakdown of organic materials by aerobicmicroorganisms. The pH range of 7.0 to 9.0 is significant as it helps to develop an environment that is neither too acidic nor too alkaline, which can be detrimental to microbial activity. Many aerobic microorganisms involved in decomposition thrive in neutral to slightly alkaline conditions. A pH below 7.0 might inhibit microbial growth and activity, slowing down the decomposition process, while a pH above 9.0 might lead to, for example, ammonia toxicity and harm the microorganisms. This specific pH range helps in maintaining a desirable environment for the microbial consortia, enhancing the efficiency of the aerobic decomposition process.

[0176] Similarly, maintaining an ORP range between -1000 mV and 1000 mV is useful for managing the redox environment of the decomposition process. ORP is a measure of the system's ability to either donate or accept electrons, which is influential in the metabolic processes of aerobic microorganisms. A positive ORP indicates an oxidizing environment, while a negative ORP signifies a reducing environment. This wide range allows for flexibility in adjusting the redox conditions to favor different stages of the aerobic decomposition process. For example, slightly positive ORP values can promote the breakdown of complex organic compounds, while values towards the lower end of the range can support other biochemical pathways relevant for nutrient recycling. The control of ORP within this range contributes to the versatility of the aerobic decomposition process, allowing for the effective breakdown of organic materials and the stabilization of the final product. In some embodiments, the ORP intervals may span from -1500 mV to 1500 mV, -1200 mV to 1200 mV, the original -1000 mV to 1000 mV, -800 mV to 800 mV, -500 mV to 500 mV, -700 mV to 700 mV, and finally, a highly specific -100 mV to 100 mV. In some embodiments, the ORP may range from -1000, - 900, -800, -700, -600, -500, -700, -700, -200, -100, 0, 100, 200, 700, 700, 500, 600, 700, 800, 900, to 1000 mV. In bioreactor operations, where the oxidation-reduction potential (ORP) plays a role in driving specific biochemical reactions, several potential alternatives across different processes can be considered. For anaerobic digestion, maintaining an ORP between -700 mV and -200 mV is typical. Aerobic digestion processes, relevant for wastewater treatment more generally, typically employ a more positive ORP range of +100 mV to +700 mV to support aerobic microorganisms in consuming organic pollutants. In the realm of nitrogen removal, nitrification may use a slightly positive ORP of +200 mV to +700 mV, whereas denitrification, which converts nitrate may operate at an ORP from -200 mV to 0 mV.Additionally, for fermentation processes or mixed culture cultivation, the ORP can vary significantly, generally within -700 mV to +700 mV, depending on the target product and involved microorganisms.

[0177] In some embodiments, the first aerobic vessel includes at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea. Pseudomonas is a genus of Gram-negative bacteria known for its metabolic versatility, allowing it to occupy different environmental niches, including soil, water, and plant surfaces. Many Pseudomonas species are beneficial for bioremediation due to their ability to degrade pollutants. Bacillus is a genus of Gram-positive, rod-shaped bacteria, widely recognized for their ability to form durable endospores, enabling them to withstand harsh conditions. Bacillus species are found in various environments, and some, like Bacillus subtilis, are used in industrial applications, including the production of enzymes and antibiotics. Escherichia, with Escherichia coli as its most well-known species, are commonly found in the intestines of warm-blooded organisms. Zoogloea is a genus of Gram-negative bacteria, notable for its role in wastewater treatment processes, forming floc that aids in the removal of contaminants from water.

[0178] In some embodiments, systems and methods include a means for transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel. Transferring is preferably done by a conduit that connects the first aerobic decomposition vessel to the second aerobic decomposition vessel. Transferring the first aerobic effluent from the initial aerobic decomposition vessel to a second aerobic vessel can be accomplished through various methods, tailored to the specific operational uses of a system designs. For example, one approach involves using air-tight hoses to ensure an oxygen-free environment during transfer, which is relevant for processes sensitive to oxygen exposure and where odor control is desired. Such methods can be automated or manually controlled, allowing for a continuous processing. Additionally, a batch process is employed depending on the facility's capacity and processing speed requirements. The air-tight transfer system is particularly beneficial in maintaining the integrity of the effluent and preventing contamination.

[0179] In some embodiments, the transfer process is designed to accommodate multiple inputs, enabling the integration of effluents from various sources into a single ormultiple secondary aerobic vessels. This approach allows for greater flexibility in handling volumes and types of effluent, optimizing the decomposition process across different streams of waste. In a batch process scenario, effluent is moved in specified amounts at scheduled or unscheduled intervals, which can help in managing the microbial health and nutrient balance within the secondary vessels. During transfer, a system can employ pumps or gravity-feed systems to facilitate the movement of effluent. The method is desirably chosen based on energy efficiency, operational costs, and the physical layout of the treatment area.

[0180] In some embodiments, the systems and methods according to the disclosure include digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent. In some embodiments, the systems and methods digest the effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8. In some embodiments, effluents from a previous stage are strategically used to maintain pH and support the complete nitrification process with no addition of other chemicals in the process. In some embodiments, the control of flow of digestate through the system is controlled by a single metering pump.

[0181] In some aspects, the techniques described herein relate to a method of producing an organic fertilizer from a wastewater or liquified organic waste, including: introducing the wastewater or liquified organic waste into an anaerobic decomposition vessel configured to produce an anaerobic effluent; wherein the anaerobic decomposition vessel includes an organic material, such as an organic material including a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained in an anaerobic condition at a pH from 6.0 to 9.0; transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel; digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent, wherein the first aerobic vessel includes at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,' transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, whichconnects the first aerobic decomposition vessel to the second aerobic decomposition vessel; digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel includes at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospircr, and filtering at least the first or second aerobic effluent or both; wherein the second aerobic effluent includes inorganic nitrogen in oxyanion form and has a carbon to nitrogen ratio greater than or equal to 3: 1, , such as 3:1, 4: 1, 5: 1, 6:1, 7: 1, or 8: 1 or more.Three Stage Description

[0182] An aspect of the disclosure is related to providing separate digestion vessels for different cultures of microorganisms. In some embodiments, some cultures of microorganisms perform different digestion processes, such as CO2 production or nitrification. In some embodiments, some cultures of microorganisms are outcompeted by other microorganisms such that certain digestion processes are reduced unless conditions are met to favor the desired cultures with the relevant digestion processes. In some embodiments, systems and methods of the disclosure provide for conditions for sensitive microorganisms, such as, for example, nitrification microorganisms, to thrive, by controlling pH, carbon / nitrogen content, and / or food sources put into the biological digestor. In some embodiments, the inputs to a digestor are controlled by performing digestions in other vessels prior to providing it to the “sensitive” digestor. In some embodiments, systems and methods of the disclosure provide for a first digestor with a pure culture of carbon decomposers in an anaerobic state, which then sends an effluent to a digestor of nitrifiers. In some embodiments, an intermediary digestor is placed between an initial anaerobic digestor and a nitrifying digestor.

[0183] In some embodiments, the techniques described herein relate to methods of producing an organic fertilizer from a wastewater or agricultural waste, such as liquified agricultural waste. In some embodiments, the systems and methods include a three-stage fertilizer production plant with specific operational characteristics and dependencies between stages. As described herein, the systems and methods may include one or more reaction vessels.

[0184] In some embodiments, the systems and methods according to the disclosure include one or more stages that are employed independently. In some embodiments, the systems and methods according to the disclosure include two or more stages that are optionally employed sequentially. For example, material may first come from Stage 1, then proceed into Stage 2, and then into Stage 3, and optionally into a stage 4. In some embodiments, a three- stage fertilizer system is configured such that the system is rate limited by the performance at Stage 1 or Stage 3, wherein Stage 2 is a rapid digestion that is facile and resilient if the Stage fails for some reason. As described herein, in some embodiments, any of the stages 1, 2, or 3 is skipped or combined with another stage.

[0185] In some embodiments, Stage 1 is an anerobic process with little or no oxygen present. In some embodiments, Stage 1 includes a vessel that is mixed by constant pumping- raw feed with fresh water fed directly into it (for example, molasses or a leafy green material with water). In a situation where the feed source is simple (e.g., containing sugars, amino acids, organic acids, etc. in an absorbable form) the digestion is rapid. These small molecules can be transported across the cellular walls and immediately be decomposed inside the cells into energy and nutrition. As a general rule of thumb, the smaller and simpler the carbon source as feed, the faster the growth of the bacteria. Large oil molecules, proteins, and starches are macromolecules or sugar polymers that require excreted enzymes for initial digestion into smaller components, which then can be absorbed into cells as a feed source. Excretion of those enzymes into the environment is expensive and the energy and carbon is lost until the larger molecules are broken down into monomers that can be absorbed. Accordingly, the initial effort to decompose complex feeds is typically slow and very energy and oxygen intensive. As the material is decomposed further the job gets easier and faster and these phenomena can be observed in the reaction rates of digesters when tuning feed rates and hydraulic retention time (how long the feed and water should “brew”).

[0186] In some embodiments, Stage 2 is an aerobic process (oxygen present and used by microorganisms to digest or react input materials). In some embodiments, the tank and plumbing for it is regularly (e.g., quarterly) cleaned with citric acid to remove buildup. In some embodiments, poor aeration into Stage 2 will slow down or cause Stage 3 to fail, and so the Stage 2 reaction vessel may have one or more ports for incorporating oxygen to maintain stable operation of Stage 2. In some embodiments, the aeration quality in Stage 2 tanks includesdissolved oxygen levels above 6ppm. In some embodiments, the aeration quality in Stage 2 tanks includes dissolved oxygen levels above 10 ppm. In some embodiments, the aeration quality in Stage 2 tanks includes dissolved oxygen levels from around 10 ppm to around 20 ppm. In some embodiments, the oxygen levels range from 6.1 to 6.5 ppm, 6.5 to 7 ppm, 7 to 7.5 ppm, or 7.5 to 8 ppm. In some embodiments, the oxygen levels are supplemented by the introduction of oxygen at a higher concentration than atmospheric. In some embodiments, medical oxygen generators or compressed pure oxygen may be used to introduced oxygen to either the gas or liquid phase. Introduction may be done through any traditional means like micro bubble aeration, introduction into circulation pump volutes for shearing and compressing, static mixers etc.

[0187] In some embodiments, Stage 3 is an aerobic process that performs a last digestion. In some embodiments, the digestion in Stage 3 is rate limited by any of the strength of the feed going into it, how established the bacteria community is, and how good the aeration is. Water from Stage 3 is filtered by a membrane machine. Run time and filtrate strength of the effluent of Stage 3 may depend on Stage 3 water quality / quantity, such that running filtration too aggressively can empty the Stage 3 vessel and cause it to fail. Similarly, in some embodiments, feeding Stage 1 too much or too rapidly can cause it to fail.

[0188] Accordingly, in some embodiments, certain indicators are used to monitor the efficiency or functioning of the multistage system. In some embodiments, the system or methods regulate the temperature at Stage 1 so that it is maintained within a range of 35 to 43 Celsius. For Stage 2, the temperature setting is flexible, optionally not requiring a specific range for successful operation. In contrast, for Stage 3, desirably the system or methods ensure that the temperature is kept between 30 to 35 Celsius.

[0189] In some embodiments, the titration levels in Stage 1 tanks are adjusted to align with the specifications outlined in the relevant report, serving as a useful indicator for maintaining chemical balance. Similarly, in some embodiments, the feeding rates in Stage 1 are managed at rates that support the maintenance or correction of Stage 1 performance. In some embodiments, the titration method used in Stage 1 tanks is based on the FOS / TAC (Free Organic Acids / Total Alkalinity Carbonate) titration. This method measures the ratio of volatile fatty acids (FOS) to the buffering capacity of the system (TAC). The FOS / TAC ratio can be an useful indicator of the digester's health, with higher ratios suggesting an accumulation ofacids and a potential risk of acidification, while lower ratios indicate a stable system. By monitoring and adjusting the FOS / TAC ratio, operators can maintain the chemical balance within the digester.

[0190] In some embodiments, in Stage 3, the dosing behavior is calibrated for a consistent and normal pulsing pattern. In some embodiments, the membrane filtration efficiency is kept at a capacity that matches the output from Stage 3, ensuring uninterrupted filtration capability.

[0191] In some embodiments, the chemical cleaning of the membranes can be implemented once every two months on a maintenance schedule. In some embodiments, the aeration quality in Stage 3 tanks includes dissolved oxygen levels above 6ppm. In some embodiments, the aeration quality in Stage 3 tanks includes dissolved oxygen levels above 10 ppm. In some embodiments, the aeration quality in Stage 3 tanks includes dissolved oxygen levels from around 10 ppm to around 20 ppm. In some embodiments, the oxygen levels range from 6.1 to 6.5 ppm, 6.5 to 7 ppm, 7 to 7.5 ppm, or 7.5 to 8 ppm. In some embodiments, the oxygen levels are supplemented by the introduction of oxygen at a higher concentration than atmospheric. In some embodiments, medical oxygen generators or compressed pure oxygen may be used to introduced oxygen to either the gas or liquid phase. Introduction may be done through any traditional means like micro bubble aeration, introduction into circulation pump volutes for shearing and compressing, static mixers etc. In some embodiments, oxygen levels of 7.5 to 8ppm represent an upper level of aeration. In some embodiments, the presence of nitrate / nitrite in Stage 2 water is avoided to improve water quality. Managing these levels prevents potential disruptions in subsequent stages and enhances the overall performance of the system. Furthermore, the production of a bulk finished fertilizer should ideally exceed the consumption needs of the greenhouses, ensuring a surplus for distribution or further use. In some embodiments, the production of a bulk finished fertilizer should ideally exceed the consumption of the greenhouses. To ensure the stability and effectiveness of the dosing pattern between stages 2 and 3, it is useful to prevent pre-acidifi cation of the feed into stage 3. Bicarbonate levels in stage 2 can be controlled to remain steady and not be pre-acidified by nitrite formation, as this may lead to an excessive feed into stage 3, disrupting the balance and potentially affecting the overall efficiency of the system.

[0192] Fig. 2 illustrates a multi-stage system 200 for treating raw organic feed to produce a filtered output for use in an irrigation system. System 200 facilitates a three-stage process for converting raw organic feed into a form suitable for irrigation use. Starting with the raw organic feed at point 201 , the material enters the Stage 1 reactor 210, where the organic feed 201 undergoes anaerobic digestion or fermentation. In this vessel, the feed is treated with fresh water to facilitate the breakdown of organic matter without the presence of oxygen. In some embodiments, raw organic feed is introduced into an anaerobic digestion / fermentation tank, where it undergoes decomposition in the absence of oxygen. Fresh water is optionally added to the system via a float valve.

[0193] In some embodiments, the product of the anaerobic digestion then flows into an ‘Aerobic Polishing’ tank, wherein oxygen is introduced, thereby reducing the chemical oxygen demand (COD) or the Biological oxygen demand of the output. Without being bound to a theory of operation, this approach will break down the organic matter further and reduce substances that can potentially harm the nitrification process in the next stage. Accordingly, the effluent material from the Stage 1 reaction 210 flows into the Stage 2 reactor 220, where the effluent is subjected to aerobic treatment. This stage may serve to polish the effluent by reducing the chemical oxygen demand, with oxygen optionally being introduced into the system.

[0194] With respect to pH, the pH will change as it moves through the system. In some embodiments, the pH in a Stage 1 digestor is 7.8 to 8.1. Without being bound to a theory of operation, this pH level is caused by the abundance of bicarbonate and ammonium. Note that in some embodiments, higher pH (more basic) is avoided due to the presence of a least small amounts of organic acid in the water. In some embodiments, the pH of the vessel is lower, if, for example, the ratio of acid to bicarbonate is high, which may in turn depend on digestor health. When the effluent is transferred to a Stage 2 digestor, the aerobic decomposition will digest the organic acids, which creates a little more CCh / bicarbonate, and as a result the solution pH rises to a higher pH, which in some embodiments is 8.3 -8.6.

[0195] The digestion continues into the Stage 3 reactor 230 through a dosing pump 221, wherein aerobic nitrification takes place, primarily by converting ammonium into nitrate. During this stage, pH will drop due to nitrification acidification, creating the primary dosing control mechanism. Since the reaction rate of nitrification is directly related to the acidproduced, the dosing of feed becomes dynamic and responsive to microbial performance. Additional oxygen may be supplied to further refine the effluent and stabilize the microorganism culture within the reactor, ensuring optimal conditions for efficient nitrification. Such a process will make the effluent more suitable for use as a fertilizer. Here, additional oxygen is optionally supplied to further refine the effluent and / or stabilize the microorganism culture within the reactor.

[0196] The next step in the process can, optionally, include a filtration subsystem 240, which separates solids 241 from the liquid effluent. The removed solids can be handled separately, while the filtrate 250 is directed toward the dewatering subsystem 260. In this phase, the liquid is split into a concentrated solution 261 and dilute water 262. The concentrated solution can be recycled back to join the filtrate, optimizing the process. In some embodiments, the systems and methods of the disclosure may not require the larger equipment to pump and move around the low-strength solutions of fertilizer as compared to the high-strength conventional compositions. Accordingly, because this method generates a stronger biodigestate and the filtrate can be concentrated with dewatering equipment, it is much more practical to implement it into existing equipment than was previously conventionally operated.

[0197] The end product of this digestion process is dilute water and a filtrate that each of which can then be channeled into an irrigation system 270, serving alternatively as a water source or a nutrient-rich water source for agricultural purposes. In some embodiments, the filtrate from Stage 3 is put into an irrigation system while the excess water is sent to a dewatering step, which uses reverse osmosis (RO) to concentrate the solids. In some embodiments, the concentrate is processed through ultrafiltration to separate the solids, which can be removed from the system, leaving behind the diluted water that cycles back into the system. In some embodiments, the system provides a closed-loop sub-system wherein water is recycled, and organic feed is converted into a nitrate-rich fertilizer through a combination of anaerobic and aerobic processes with intermediate filtration steps to create a final product for use in irrigation.Methods for cultivating microorganisms for different decomposition pathways.

[0198] Without being bound to a theory of operation, the following description provides rationales and solutions to issues with switching to organic fertilizers. In someembodiments, a goal for the multistage bioreactors includes performing decomposition on the biomaterials to the point where the plants can absorb the nutrition but without system failures, such as excess microorganism detritus clogging an irrigation system. In some embodiments, this decomposition includes mineralizing, which refers to decomposing organic material to a more stable mineral form, such as salts of potassium, magnesium, iron, ammonia, or nitrate.

[0199] Traditional methods for mineralizing organic feeds can use activated sludge. Activated sludge is an aerated body of water and bacteria, and in these systems most mineralization can occur in one digestor via aerobic bacteria. Activated sludge depends on two groups of bacteria primarily - heterotrophic and autotrophic nitrifiers. One difference that separates these bacterium types is their energy and carbon source. Heterotrophic bacteria usually depend on the energy available primarily in carbon-to-carbon bonds in complex organic molecules. As those bonds are broken, they utilize the carbon in those compounds for their own cells and then any surplus carbon is discarded as CO2. Heterotrophic bacteria are aerobic and use oxygen for respiration. Some species of Heterotrophic bacteria can also survive under microaerophilic conditions, surviving right on the edge of no oxygen. The growth rate of heterotrophic bacteria is comparably fast to other microbes. While heterotrophic bacteria aggressively consume carbonaceous feeds, these microorganisms also produce a substantial quantity of waste. Most notably the wastes are heat, CO2, ammonium, all other mineral ions (potassium, magnesium, calcium, etc.), and dead cells. Under some conditions, half of the carbon fed to the bacteria is converted to CO2, while the other half is converted to cell material. This results in massive dead cell waste and poor return of CO2, which, as described herein, can create incompatibilities with irrigation systems.

[0200] Another relevant waste product is the ammonium. Ammonium is the result of nitrogenous feed decomposition and is not a particularly well decomposed state for nitrogen. Plants are able to use ammonium as a nitrogen source very effectively, but in some crop production ammonia is only present in minor amounts relative to the other primary nitrogen sources of nitrites and nitrates. Accordingly, in some embodiments, systems and methods of the disclosure digest nitrogen and ammonia further into nitrite / nitrate. Autotrophic bacteria, and, for example, specifically chemoautotrophic bacteria can perform this digestion. These bacteria generally depend on the energy of the oxidation of ammonium into a higher oxidation state. As autotrophic bacteria typically do not use carbonaceous feeds for energy, autotrophicbacteria will also rely on other sources for carbon assimilation such as CO2 in water as a carbon source, in the form of aqueous bicarbonate / carbonate.

[0201] As CO2 and ammonium are waste products of their heterotrophic counterparts, autotrophic bacteria have specialized in the niche of utilizing these two resources, but these oxidation reactions are very low energy yielding and the oxidation of ammonium also creates acid, which can drive CO2 out of water. Accordingly, these autotrophic bacteria “Nitrifiers” are very poor competitors in the microbial space because autotrophic bacteria generally do not grow very quickly as their energy and carbon sources are typically rare and depleted. However, as provided by the disclosure, certain conditions allow the autotrophic bacteria to perform well, under both selective water chemistries and competition.

[0202] In some embodiments, systems and methods provide for microorganisms that can perform Ammonium oxidation and Nitrite Oxidation. In some embodiments, these reactions is carried out by two different bacteria strains: Nitrosomonas and Nitrobacter, which predominantly perform Ammonium oxidation and Nitrite Oxidation, respectively. Note, bacteria species may vary and may perform both reactions, like, for example, Nitrospira, which performs both. By way of one example only, these oxidation processes of the Nitrosomonas / Nitrobacter model are illustrative of some of the typical chemistry in the bioreactors. In a mixed culture of bacteria in activated sludge, the population of these chemoautotrophs is 3-7%, while 93-97% is heterotrophs. Being slow growing and easily outcompeted, they occupy a small proportion of the biomass and are easily wiped out with overfeeding of heterotrophs, like when carbonaceous feed is loaded in rapidly. The activated sludge systems described above might convert half of the carbonaceous feed delivered to a digestor into microbial cell mass with the other half will be eliminated as CO2 gas. In contrast, the carbon fraction fed to an anaerobic digestor culture including chemoautotrophs as described herein might convert 5% of cell mass while the other 95% of the carbon fraction is eliminated as CO2 and methane. This presents an opportunity when designing an efficient digestor system, which may also produce high strength effluent.

[0203] In some embodiments, systems and methods provide for a fertilizer production process that yields Nitrate and other accompanying minerals from a complex feed, and which may also minimize waste to microbial biomass along the way.

[0204] Accordingly, in some embodiments, systems and methods culture both organisms that break down the complex carbonaceous feed, and nitrification microorganisms that are sensitive or otherwise slow growing. In some embodiments, nitrification microorganisms have two main inputs, ammonium and carbon dioxide (in bicarbonate form). In some embodiments, a strong culture of nitrifiers is cultured by consistently feeding them these two ingredients. In some embodiments, other supportive minerals (potassium, calcium, magnesium, iron, etc.) would also not be limited. As the nitrifiers use bicarbonate not just for the reactions, but also their cellular growth, providing higher amounts of bicarbonate, than mere stoichiometric amounts for ammonium oxidation and nitrite oxidation, can led to more robust growth of the nitrification microorganisms.

[0205] In some embodiments, an excess amount of bicarbonate is provided. Where a theoretical ratio might recommend (7.5: 1) bicarbonate to ammonium, some embodiments of the disclosure provide for ratios (8.6: 1). In some embodiments, the ratio of bicarbonate to ammonium is one of the following: 7.6:1, 7.7:1, 7.8:1, 7.9:1, 8.0:1, 8.1 :1, 8.2:1, 8.3:1, 8.4:1, 8.5: 1, 8.6: 1, 8.7: 1, 8.8:1, or 8.9: 1.

[0206] Some embodiments of the disclosure include a digestor of nitrifiers. In some embodiments, a digestor of nitrifiers is provided a feed of bicarbonate to ammonium of 8.6: 1, wherein such digesters have been observed to digest to completion and the pH will be stable at 6. In some embodiments, the disclosure provides for large amounts of CO2 released from initial digestion processes to finish oxidizing ammonium to nitrate. In some embodiments, the disclosure provides for efficient methods for extracting CO2 from feed. In some embodiments, the disclosure provides for using high carbon: nitrogen ratio feeds to benefit nitrification microorganisms. In some embodiments, the ratio of carbonmitrogen is one of the following: 10: 1, 9: 1, 8: 1, 7: 1, 6:1, 5:1, 4:1, 3:1, 2:1, or 1 :1. In some embodiments, both efficient methods for extracting CO2 from feed and additional sources of high carbonmitrogen ratio feeds are used.

[0207] In some embodiments, raw feeds are digested by anaerobic digestion. In some embodiments, aerobic heterotrophic microbes are used to digest the raw feeds. Anaerobic digesters generally perform digestion via excreted enzymes that are capable of breaking down highly complex compounds and mixtures. Also, in contrast to aerobic oxidative decomposition, anaerobic decomposition is generally not limited to the oxidizing potential ofoxygen, rather the reductive processes have the potential to decompose strong bonds such as the disulfide bridges in keratin. In some embodiments, the yield of CO2 from anaerobic digestion is higher, as compared to heterotrophic microbes. In some embodiments, the carbon loss to microbial cells is less for anaerobic digestion than with heterotrophic microbes. In some embodiments, the byproducts of anaerobic digestions are ammonium, CO2, and mineral ions. In some embodiments, additional products are produced including organic acids and methane gas (if, for example, operating under methanogenic conditions).

[0208] Some embodiments use an anaerobic culture as a preliminary digestor, whereby later stages of the digestion receive the end product of the anaerobic digestate. If the later stages included nitrifying cultures, these potentially sensitive cultures would have free rein on the entirety of the culture since the anaerobic cells would rapidly degrade in the presence of oxygen. Secondly, providing anaerobic feed to nitrifiers according to the disclosure is preferable because of the low loss of carbon to cell volume, production of ammonium, and the naturally highly enriched solution in CO2.

[0209] In some embodiments, a system is provided with an intermediary vessel that removes the opportunity for heterotrophs to destroy the nitrifier culture in the event that feeding loads are too high. Similar to competition in an aerobic environment, nitrifiers can be outcompeted by aerobic heterotrophs when the aerobic heterotrophs have plenty of feed. Accordingly, in some embodiments, intermediary digesters or methods to complete digestion by, for example, adding additional nutrients or feed to allow the digestor to run to completion are utilized.Stage 1 Anaerobic Polishing - pre-nitrification cleanse

[0210] As described above, in some embodiments, methods and systems according to the disclosure include an anerobic process under little or no oxygen. In some embodiments, this process is a preliminary step and includes a vessel that is mixed by constant pumping- raw feed with fresh water fed directly into it (for example, molasses or a leafy green mixture with water). In some embodiments, anaerobic digesters are operated under acidic conditions. Acidic operation generally refers to cultures enriched for growth of the acid forming bacteria, those which decompose a complex feed into small organic acids. These acids are typically veryunpleasant smelling and include acids like butyric acids (rotten egg) and / or acetic acid (vinegar).

[0211] Fortunately, under anaerobic conditions there is still another group of bacteria that like to digest those acids and convert them into methane and CO2. In some embodiments, anaerobic digesters are operated under methanogenic conditions. In some embodiments, both acidic and methanogenic cultures are provided, in one or more vessels. In some embodiments, these bacteria decompose these organic acids to methane and CO2 that is used in downstream processes. Note that in some embodiments, such as, for example, in a continuous flow culture of anaerobes, the water taken out of the reactor may still have residual organic acid in it. In some embodiments, the residual organic acid presence is controlled depending on the health and feed of those methanogens.Stage 2

[0212] As described above, one aspect of the disclosure is directed to vessels / cultures that decrease the number of specific types of nutrition that would allow undesirable microorganisms to outcompete desired organisms. In some embodiments, systems and methods provide for nutrients specific for nitrifiers, and avoid stimulating the growth of heterotrophs in the same vessel as the nitrifiers. In some embodiments, the growth of heterotrophs is stimulated in a precursor vessel in order to eliminate the nutrients for heterotrophs. In some embodiments, excess carbonaceous feed is avoided in the nitrifier vessel(s), so as to avoid stimulating the growth of heterotrophs.

[0213] By way of example, nitrifiers can be fed directly from an anaerobic digestor as the remaining organic acids can stimulate heterotrophic growth. Organic acids are desirable heterotroph feed because these small molecules are highly soluble and can readily be absorbed into the cells. Accordingly, when the acids are introduced into an aerobic, aerated environment, the heterotrophic culture will burst in population -and may outcompete nitrifiers. Thus, in some embodiments, the anaerobic digestate is fed directly to a small culture of aerobic heterotrophs, which gives them a competitive advantage to such a degree that the nitrifiers will not survive, then once the organic acids are all consumed, the resultant product is introduced into a nitrifier digestor. This general process may be referred to herein as aerobic polishing.

[0214] Accordingly, in some embodiments, the systems and methods provide an intermediate digestor that has mild aeration such that aggressive heterotrophs will consume the majority of organic acids. In some embodiments, the heterotrophs aggressively consume organic acids, but are also stressed by providing limited amounts of oxygen. In some embodiments, the oxygen levels are limited, and is too low to sustain nitrifiers (for example, less than 2ppm O2 and typically near 0 and the environment becomes so competitive that only heterotrophs will survive. Note that some nitrification bacteria might monitor their environment for excess free carbon feed which would indicate high levels of competition, and if the carbon is too high, they choose to not grow. Accordingly, some embodiments of the disclosure are directed to reducing the amount of excess free carbon entering the nitrifying vessel.

[0215] In some embodiments, an effluent exiting a vessel undergoing this “aerobic polish” comprises a partially processed biomaterial that has had most of the free organic acids removed, CO2 generated from the process, ammonium, bicarbonate, minerals, and dead cells. This feed has desirable characteristics for a nitrification reactor.

[0216] In some embodiments, the culture in the intermediate vessel, or Stage 2 vessel, is controlled by monitoring the ORP of the vessel and introducing oxygen or air when the oxidation reduction potential deviates from desirable levels. By way of example, in principle, Stage 2 is a well aerated digestor that if left alone, would naturally begin to foster nitrification bacteria. If a Stage 2 vessel were to begin nitrifying, the pH might drop and the dosing control into Stage 3 will be impacted negatively. In a typical example, a drop in pH will cause the dosing from Stage 2 to Stage 3 to be much too rapid, or not work at all. Accordingly, in some embodiments, systems and methods are provided that limit polishing in the intermediate vessel and prevent the growth of nitrification microorganisms.

[0217] In some embodiments, control over the intermediate vessel conditions is provided by maintaining a consistent feed of anerobic digestate (for example, from Stage 1) and maintaining moderate aeration. If, for example, an insufficient amount of aeration is provided to the intermediate tank, an undesirably high amount of undigested acids is generated in the effluent, such that a nitrifying tank would then become a host to other microorganisms that can outcompete the nitrifying microorganisms.

[0218] In some embodiments, an ORP measurement (Oxidation-Reduction Potential) is used to measure how far into the Oxidation cycle the digestion has gone. The Oxidation-Reduction Potential is a measurement of the oxidation or reduction voltage that exists in a solution. Solutions with high dissolved oxygen will have a commensurately high ORP measurement (+1500mV). Inversely, low oxygen environments will have a low ORP (- 600 to -2000 mV). Note that Oxygen is not the only species that contributes to the ORP. The Nitrification cycle, as an example, also has a change in oxidation state of nitrogen through its series of transformations.

[0219] Accordingly, some embodiments keep a Stage 2 reactor at an ideal state, with suitable acid removal, but no or limited nitrification. In some embodiments, intermediate vessels have an ORP of OmV + / - 150mV. In some embodiments, intermediate vessels have an ORP of OmV + / - lOOmV. In some embodiments, intermediate vessels have an ORP of OmV + / - 50mV. Note that the ORP measurement may vary between solutions. For example, one day the desired conditions in a reactor may correspond with an ORP of 0 mV, but then the next day the ORP could be, for example, +50 mV due to a number of factors, such as changes in the conductivity of the solution or another factor. Additionally, as would be expected by one of skill in the art, different sensors can also yield different results, but generally similar solutions will have relatively close measurements.

[0220] However, in some embodiments, an ORP of 0 mV within + / - lOOmV is a desirable ORP for operating conditions of the reaction vessel. In some embodiments, when the ORP deviates from the desired amount the methods and system provide for modulating the aeration rate. In some embodiments, a system comprises one or more installed valves on the air supply lines into a Stage 2 tank. In some embodiments, ORP sensors are installed on the internal recirculation loops of the Stage 2 tanks and the aeration is turned on and off to keep the ORP within 25mV of a setpoint. In some embodiments, a setpoint is within 150mV of 0 mV. In some embodiments, the setpoint is 150 mV. In some embodiments, the setpoint is -150 mV. In some embodiments, the setpoint is 100 mV. In some embodiments, the setpoint is -100 mV.

[0221] In some embodiments, the transfer of effluent from, for example, Stage 2 to Stage 3 is controlled by a positive displacement dosing pump. In some embodiments, this transfer is controlled using a controller software. In some embodiments, a target pH is set inthe control software and when reached, the system will move effluent, pulse-by-pulse, to maintain characteristics, such as a pH range, in, for example, a Stage 3 reactor.

[0222] An aspect of the disclosure is directed towards the transfer of effluent between reaction vessels to create properly balanced chemistry. For some applications, bioreactors are limited in their access to refined chemicals to control water pH, etc. In such embodiments, the disclosure provides for systems and methods that create desired chemistries based on the initial inputs to create properly balanced chemistry throughout the process. For example, in some embodiments, instead of adding a pH controlling additives (e.g. liming chemistry, like potassium bicarbonate,) the systems and methods control the progression of inputs to generate a high pH dig estate, such as a digestate coming from Stage 2 into Stage 3. In some embodiments, the pH may be within any of the following ranges: 5-13 pH, 5-12 pH, 5-1 IpH, 5-10 pH, 6-10 pH, 6-9 pH, or 6-8 pH.

[0223] In some embodiments, a single point of control can be used to control the transfer of liquid between Stage 2 to Stage 3. This configuration would have some benefits to control the system because it is positioned at what is typically the rate-limiting step and may also use pH as a feedback measurement, which makes for simple operation. In some embodiments, movement of water from Stage 1 to Stage 2 is controlled via a float valve. By way of example, as water is removed from Stage 2, water is automatically moved from Stage 1 to Stage 2 to refill it. In some embodiments, fresh water is used to refill Stage 1, so Stage 1 may also include one or more float valves that supply fresh water to Stage 1.

[0224] Accordingly, in some embodiments, pH is controlled by using a Stage 2 effluent as a source of a high pH solution that is fed into a nitrification reactor. Adjusting pH, as described further herein, will modulate and maintain balance between ammonia and nitrite oxidation activity, which will reduce the pH as nitrogen species are oxidized by the cultures in the reactor. In some embodiments, a desired pH setting and dosing speed / reaction behavior are set, and the whole system is run continuously when provided a proper supply of raw feed and water into Stage 1, thereby maintaining the overall bioreactors’ health. In some embodiments, an aqueous ammonium solution may be added to any one of the digesters as a pH control agent and / or as an additional source of nitrogen. In some embodiments, an aqueous ammonium solution may be derived from pig manure. In some embodiments, an aqueousammonium solution may be particularly beneficial in stages 2 or 3 of the process described herein.Stage 3

[0225] An aspect of the disclosure is directed to a vessel that receives feed through an anaerobic digestor, a polishing digestor, and then to a nitrification reactor. Note that this step might be referred to as a reactor / digestor interchangeably because this stage of the process might not be strictly decomposing and digesting, but instead “reacting” (via bacteria) inorganic ions with oxygen to extract the final and last bit of energy available with chemoautotrophs. In some embodiments, the systems and methods according to the disclosure provide for a vessel to digest ammonium (NH4+) into Nitrite (NO2-) and Nitrite (NO2-) to Nitrate (NO3-).

[0226] For purposes of illustration, and without being bound to a theory of operation, the following describes differences in the biochemistry of the microorganisms in, for example, a Stage 3 reactor. Heterotrophic growth on sucrose, for example, has high growth rates due in part to the large energy yields from digesting sucrose. Ammonia oxidizing bacteria (AOB) on the other hand receive half as much energy per reaction and the last in the chain, the nitrite oxidizing bacteria (NOB), yield 5 times less energy than the AOB.

[0227] Additionally, in an AOB reaction, 4 protons (H+) are produced when an ammonia molecule is decomposed, 4 acid units are produced. Accordingly, in some embodiments, the bicarbonate from previous reactions is useful in maintaining the solution pH in the ideal range. Secondly, as this reaction proceeds the microbial cells use carbon to build their cell walls and organelles - this too can be sourced by the bicarbonate. About 15% of the bicarbonate supply might go to cell production of both AOB and NOB.

[0228] Some embodiments provide for the oxidation of nitrite available for NOB. The NOB reaction itself does not strictly yield acid, however bicarbonate is still consumed to support cellular respiration and growth. Taking into account the 5X lower energy yield from the NOB reaction, these microorganisms have a lower competitive advantage over AOB when considering energy yield alone. Other desirable features such as improved biofilm stability provide adapted mechanisms to maintain a competitive advantage. However, the initial establishment of the culture can be challenging. Accordingly, some embodiments of the disclosure provide for methods of establishing such cultures through inoculum.

[0229] In some embodiments, a system will employ pH level as a control variable to modulate the use of the high alkalinity-to-nitrogen ratio solution produced in the initial stages. Some embodiments will ensure that the pH remains within a predetermined range. By adjusting the feed from Stage 2 to Stage 3 based on the pH level. In some embodiments, the process in Stage 3 will acidify the solution, as compared with the alkaline conditions fostered in Stages 1 and 2. In some embodiments, Stage 3 serves as a bottleneck in the overall process due to its rate-limiting nature, such that monitoring and adjusting the feed based on pH changes, which are indicative of the reaction rate, is useful. As the pH decreases due to the ongoing digestion process, the system can be compensated by increasing the feed, thereby maintaining a desired reaction pace.

[0230] Additionally, the strategy for maintaining a balanced culture of ammonia oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) within the Stage 3 tank can require fluctuating the pH approximately 0.5 units above and below a setpoint of 5.8. This method purposely subjects the faster-growing AOB to stress with lower pH levels but allows for their recovery as the pH increases. Conversely, NOB can tolerate the full pH spectrum, enabling them to remain competitive. This approach allows for a coexistence of both bacterium types, which improves the system’s efficiency.

[0231] In some embodiments, the inclusion of chemoautotrophic microorganisms, such as those from the Nitrosomonas, Nitrobacter, and related genera, along with the filtration of aerobic effluents, will further enhance the systems’ performance by ensuring the presence of inorganic nitrogen in the effluent and maintaining a desirable carbon to nitrogen ratio. Addressing the challenge of transporting low-concentration liquid digestates, systems may employ nanofiltration and membrane technologies like Reverse Osmosis (RO) and Forward Osmosis (FO) for solid removal and direct dewatering. In some embodiments, this process will significantly concentrate the material, thereby reducing the need for extensive storage space and making on-site storage more feasible and efficient.

[0232] In various embodiments, the features and functionalities described herein with respect to the base media contactor, the control regime for hydraulic retention, solids loading, and microbial activity modulation may be implemented in conjunction with any of the other features disclosed throughout this application. The system elements described in relation to base media contactors — including, but not limited to, the modulation of pH via carbonate oroxide dissolution, the release of agronomically beneficial mineral ions, the pre-filtration of particulates in downflow configurations, and the closed-loop integration with pH and nitrite monitoring in aerobic nitrification stages — are compatible with and may be functionally combined with the multi-stage digestion processes, the microbial control strategies, the nutrient dosing approaches, and the monitoring and feedback frameworks described elsewhere in the application.

[0233] In some embodiments, the integration of the base media contactor may be implemented in systems comprising at least one anaerobic decomposition vessel (Stage 1) and at least two aerobic decomposition vessels (Stages 2 and 3), wherein fluid transfer, aeration levels, and substrate dosing are regulated in real time based on operational indicators such as FOS / TAC ratios, electrical conductivity (EC), ammonium and nitrite concentrations, bicarbonate availability, and pH dynamics. In certain embodiments, the base media contactor is fluidically coupled downstream of Stage 2 and upstream of Stage 3, such that effluent undergoing nitrification in Stage 3 has first been conditioned through partial alkalinization, mineralization, and / or pre-filtration.

[0234] In some embodiments, the base media contactor may be used in systems that employ controlled pH cycling within Stage 3. In such cases, the media contactor provides a responsive buffer capacity, mitigating microbial stress associated with nitrite accumulation and acidification. The dissolution of alkaline-reactive solids is further used to restore or maintain optimal nitrifying microbial function, and can be timed to coincide with reduced aeration strategies or extended retention within Stage 2 to avoid over-oxidation or ammonia stripping.

[0235] The features of the base media contactor are also combinable with systems that manage solids loading in the anaerobic vessel (Stage 1). In some embodiments, a base media contactor may be combinable with systems that manage solids loading in the anaerobic vessel proportionally to downstream demand, and thereby, for example, maintaining steadystate ionic strength. Some embodiments may avoid substrate overload or ammonium toxicity. In some embodiments, the mineral supplementation provided by the base media contactor compensates for ionic imbalances downstream. In some embodiments, the mineral supplementation provided by the base media contactor supports biofertilizer nutrient profiles compliant with organic certification standards.

[0236] Furthermore, it is specifically envisioned that the base media contactor may be combined with embodiments that employ staged filtration — including microfilters, ultrafilters, or nanofilters — the pre-filtration function of a downflow base media contactor reduces fouling potential and extends membrane service life. In some embodiments, the physical function operates concurrently with the chemical functions of alkalinity and nutrient ion supplementation. In some embodiments, the base media contactor may operate as an integrated pretreatment unit process that complements broader system stability and continuous operation.

[0237] Accordingly, all of the system elements described herein — including aeration modulation, substrate dosing strategies, microbial monitoring, mineral supplementation, hydraulic retention control, and multi-stage digestion — may be variously combined with the use of the base media contactor as described, whether operated in upflow or downflow configuration. These combined implementations offer synergistic performance, including improved microbial resilience, reduced process variability, enhanced nutrient recovery, and stable biofertilizer composition. The descriptions provided herein are intended to support claims that recite such combinations and all the permutations therein.Characterization of Effluents and Filtrates

[0238] The functioning of the microbial digestion system may be evaluated through various effluent metrics. These metrics provide useful insights into the efficiency and stability of the digestion process. Certain parameters such as Biochemical Oxygen Demand (BOD), Volatile Acids, Chemical Oxygen Demand (COD), pH, Ammoniacal Nitrogen, Nitrite Nitrogen, and Nitrate Nitrogen may be measured at different stages of the process. The system's performance may be assessed by analyzing this data points. The following section presents a summary of the effluent data, and highlights relevant changes observed across different stages, as well as the effectiveness of specific treatments, such as the addition of humic acids to membrane filtrate products.

[0239] The table below shows the measurements taken at different stages of the microbial digestion process and the effects of adding humic acids to the membrane filtrate products. The data include Biochemical Oxygen Demand (BOD), Volatile Acids, Chemical Oxygen Demand (COD), pH, Ammoniacal Nitrogen, Nitrite Nitrogen, and Nitrate Nitrogen.The table illustrates the changes in these parameters from Stage 1 to Stage 3, and the characteristics of two membrane filtrate products with added humic acids. The Chemical Oxygen Demand (COD) increased dramatically, and the total organic carbon (TOC) raised significantly, as expected. However, the Biochemical Oxygen Demand (BOD) stayed low, which was the desired result. This demonstrates the addition of a stable carbon source that has a low BOD but raises the C:N ratio effectively.Table 1 : Chemistry of stages 1. 2. and 3 of the treatment processUnits in mg / LBacterial Strain Disclosure and Characterization

[0240] Fig. 4 shows a pie chart summarizing the microorganisms by class, for the microorganisms measured to be present in one example of a stage 1 reactor. The pie chart indicates the relative proportions of different classes of bacteria or other microorganisms by the relative size of the segments in the pie chart. The composition represented in this specific pie chart does not imply that all configurations within the bioreactor must adhere to this particular distribution. The composition of bacteria in an anaerobic bioreactor that is processing biological waste from a farm can give us some insight into the likely outputs of the process. The largest segment of the chart is corresponds to "Chloroflexia" , which constitutes 29.37% of the chart. This is followed by a " is the next significant group at 13.12%, and " Bacteroidia" follows at 10.18%. There are smaller segments for " Synergistia" (4.14%) and "Bacilli" (1.27%). Other classes are labeled in the legend but are not specifically labeled in the chart, as these microorganisms make up smaller fractions of the total. These microorganismsinclude "Unclassified" and various other classes such as " Coriobacteriia" , " Planctomycetia” , ” Art ri bacteria” , and several others, down to " Flavobacteriia" at the end of the legend list.

[0241] Given that the dominant class of bacteria is Chloroflexia, which are present in various anaerobic environments, including sewage sludge and lake, their presence in a high proportion suggests that the reactor might be operating in conditions that favor their growth.

[0242] The relatively large presence of Clostridia is noteworthy because members of this class are known for their role in anaerobic digestion, where they break down complex organic molecules into simpler compounds, such as organic acids. In anaerobic bioreactors, these bacteria are often responsible for the hydrolysis and acidogenesis stages, where large polymers like carbohydrates, proteins, and fats are broken down into sugars, amino acids, and fatty acids, and then further into products like hydrogen, carbon dioxide, and organic acids. Actinomycetia, another class with substantial representation in the bioreactor, are also capable of degrading complex organic substances and may contribute to the production of similar intermediate degradation products. Deferribacteres and Bacteroidia are also associated with the degradation of complex organic molecules under anaerobic conditions. Their activities could enhance the breakdown process, contributing to the overall efficiency of the bioreactor.

[0243] Accordingly, an output from such a bioreactor, considering the bacterial composition and the anaerobic conditions, may be biogas, a mixture of methane (CH4) and carbon dioxide (CO2), with methane being the desired product for energy generation. Alternative outputs include organic acids, including a mixture of volatile fatty acids (VFAs) as intermediary products of the digestion process. The proportions of these outputs may depend on the operational conditions of the bioreactor, including temperature, pH, retention time, and the specifics of the biological waste input.

[0244] Fig. 5 shows a pie chart summarizing the proportions of different bacterial classes, in a system of bioreactors’ second stage, where the effluent from the previous stage is introduced into the instant bioreactor. A large class in this stage is "Actinomycetia” , which occupies a large portion of the pie chart, labeled at 35.57%. The next significant class is "Clostridia” at 27.67%, followed by " Betaproteobacteria” at 11.24% and "Deferribacteres” at 9.52%. "Chloroflexia” is present at 4.49%, " Synergistia” at 3.96%, "Bacteroidia” at 3.11%, and "Bacilli" at 0.7%. Similar to Fig. 4, other classes are noted in the legend, indicating their presence in smaller amounts not clearly distinguishable in the pie chart. These include" Alphaproteobacteria" , "Unclassified", " Coriobacteriia" , " Deltaproteobacteria" ,'' Thermo / eophi / ia'', and several others, ending with "Acidobacteria" at the bottom of the legend.

[0245] Fig. 5 indicates a shift in bacterial dominance from "Chloroflexia" in Stage1 to "Actinomycetia" in Stage 2, which is also indicative of the conditions within the bioreactor or the stage of waste processing. The increase in the relative abundance of Actinomycetia from Stage 1 to Stage 2, alongside a notable presence of Clostridia, the transition from strictly anaerobic to semi-aerobic or aerobic conditions. Actinomycetia includes a diverse group of bacteria, many of which are facultative aerobes or obligate aerobes, thriving in the presence of oxygen. They are known for their ability to break down a wide array of organic compounds and are often found in soil and composting systems where oxygen is more available. Introducing aerobic conditions can activate and accelerate the metabolic activity of Actinomycetia, thereby increasing their proportion in Stage 2.

[0246] Note that in some embodiments, the differences between stage 1 and stage2 could be created by introducing a different inoculum in Stage 2. For example, if the inoculum contains a high concentration of Actinomycetia and other aerobes, these organisms would outcompete anaerobes when oxygen is introduced, leading to the observed dominance in Stage 2. This shift can also be the result of the designed progression in the bioreactor stages, where Stage 1 focuses on reducing the mass and complexity of the organic waste under anaerobic conditions, and Stage 2 aims at further breakdown and stabilization of the remaining matter under aerobic conditions.

[0247] Other factors that may be used to induce a shift in the composition in the reactor include changes in pH, temperature, or the addition of specific nutrients or substrates that favor the growth of, for example, Actinomycetia or other aerobes. Adjusting these parameters can promote the growth of different microbial communities between stages. For example, a higher temperature could promote thermophilic bacteria in one stage over another, or a change in pH could select for bacterial groups that prefer neutral or slightly alkaline conditions.

[0248] Fig. 6 shows a pie chart summarizing the proportions of different bacterial classes, in a system of bioreactors’ third stage. A large class in this stage is 'Actinomycetia," which constitutes 36.41% of the chart, slightly increasing from the 35.57% in Stage 2." Flavobacteria" appear in Stage 3 at 17.11%, which is a new addition to the major classes compared to the previous stages. Other notable classes include "Betaproteobacteria" at 10.39%, " Alphaproteobacteria" at 5.72%, " Sphingobacteria" at 7.2%, and "Clostridia" at 6.88%. Additionally, there are smaller proportions for "Cytophagia" (4.87%), " Synergistia" (1.61%), and " Coriobacteriia" (0.61%), with "Unclassified" making up 3.65%.

[0249] Comparing Stage 3 to Stage 2, there is a slight increase in the proportion of Actinomycetia, which is consistent with continuing semi-aerobic or aerobic conditions. The appearance of Flavobacteria as a major class is notable for their role in the degradation of complex organic compounds, and their increase is consistent with a shift in the operational conditions that favor their growth, such as changes in nutrient availability, pH, temperature, and dissolved oxygen levels.

[0250] The presence and proportions of other classes like Betaproteobacteria and Sphingobacteria suggest that the bioreactor maintains a diverse microbial community, which is useful for the comprehensive breakdown of organic matter. The decline in Clostridia from Stage 2 to Stage 3 could be due to the shift towards more aerobic conditions, as Clostridia are typically anaerobic.

[0251] The disclosure also provides for methods of cultivating specific strains or mixtures thereof. To cultivate a specific strain of bacteria, such as Comamonas, in significant amounts, starting from a small inoculum in a stage 3 tank of a wastewater treatment process, several methods can be employed, each tailored to optimize growth conditions and effectively scale up the culture.

[0252] One approach is batch culturing, where a small inoculum of Comamonas is isolated from a mixed microbial community or obtained from a microbial culture collection. The growth medium may be optimized for Comamonas, typically including appropriate sources of carbon and nitrogen that the bacteria can metabolize. One may adjust the pH and maintain adequate oxygen levels, as Comamonas thrives under aerobic conditions. The culture may be kept at optimal temperatures, generally between 20°C and 30°C, and scaled up progressively by transferring the culture to larger tanks while maintaining consistent environmental and nutritional conditions.

[0253] Another method is continuous culture, which involves setting up a bioreactor system such as a chemostat or turbidostat. In this system, new medium iscontinuously fed into the culture vessel, and culture fluid containing microorganisms is continuously removed to maintain a steady state of bacterial growth. This method allows for the continuous provision of nutrients and removal of wastes, which helps maintain optimal growth conditions. Parameters like flow rate, nutrient concentration, and dilution rate can be finely controlled to ensure the culture remains in its growth phase.

[0254] The fed-batch culture technique combines elements of batch and continuous cultures by starting with a batch culture and then intermittently adding fresh nutrient medium without removing the culture fluid. This approach allows higher cell densities by preventing nutrient depletion and toxic buildup. The timing and amount of nutrients added are managed to optimize the growth environment. Using selective media can also enhance the growth of Comamonas by inhibiting competing microbes. This involves adding specific substrates or inhibitors that target metabolic pathways unique to Comamonas or closely related strains.

[0255] Table 2 below lists the various bacterial species included in stages 1, 2, and 3 of the treatment process. This disclosure outlines a composition that comprises a mixture of bacteria, specifically a mixture of isolated bacteria selected from the group consisting of at least 7, at least 6, at least 5, at least 4, and at least 3 of the bacterial strains presented in the table. This allows for flexibility in configuring the microbial composition to meet specific bioreactor conditions and treatment objectives. Table 2 is sorted first by the largest proportion of microorganisms in stage 1, then by the largest proportion in stage 2, and finally by the largest proportion in stage 3. As can be observed in the first rows of the table, the originally high concentration of Chloroflexi decreases from stage 1 to stage 2, but is not completely eliminated, which is consistent with a multistage reactor that allows some cell detritus from an earlier stage to progress to subsequent stage bioreactors.Table 2: Bacterial species included in stages 1, 2, and 3 of the treatment process

[0256] An aspect of the disclosure provides for bacterial types with their respective percentages for each of the three stages of treatment. For Stage 1, the composition may include Chloroflexia from the order Chloroflexales, which may be present at approximately 30% of the bacterial population. Clostridia, belonging to the phylum Firmicutes, may make up a value within + / - 5% of 20% of the mixture. Actinomycetia from the family Actinomycetaceae, genus Actinomyces, and order Actinomycetales of the phylum Actinobacteria, account for a value within + / - 5% of 15%. Deferribacteres may be present at a value within + / - 5% of 15% and Bacteroidia from the family Rikenellaceae and order Bacteroidales of the phylum Bacteroidota may be present at a value within + / - 5% of 5%.

[0257] In Stage 2, Actinomycetia of the same classification as in Stage 1 may be a value within + / - 5% of 30%, Clostridia may be present at a value within + / - 5% of 25% and Deferribacteres at a value within + / - 5% of 10%, Chloroflexia, similar to those in Stage 1, may account for a value within + / - 5% of 5% and Bacteroidia may be at a value within + / - 5% of 3%. For Stage 3, the composition may include Actinomycetia at a value within + / - 5% of 30%, Clostridia at a value within + / - 5% of 5%, Chloroflexia at a value within + / - 5% of 1%, and Deferribacteres at a value within + / - 5% of 0.5%.

[0258] In some embodiments, at least one of the anaerobic decomposition vessel, the first aerobic vessel, or the second aerobic vessel includes a microorganism culture derived from an inoculum, such as, for example, worm castings and / or crop waste.

[0259] In some embodiments, the anaerobic decomposition vessel includes worm castings, and the worm castings include a composition of microorganisms capable of digesting the organic material into organic acids under basic conditions, said composition of microorganisms including bacteria known for their ability to decompose complex organic materials into simpler molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter.

[0260] In some embodiments, the anaerobic decomposition vessel includes at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Escherichia coli, Escherichia fergusonii, Escherichia albertii, Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae.

[0261] In some embodiments, the anaerobic decomposition vessel includes a plurality of microorganisms, wherein the majority of the microorganisms are carbon composing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria.

[0262] In some embodiments, the first aerobic decomposition vessel includes facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of Lactobacillus, Streptococcus, and Enterobacter.

[0263] In some embodiments, the first aerobic decomposition vessel includes obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, e.g., species from the genera Bacteroides, Clostridium, or Eubacterium.

[0264] In some embodiments, the first aerobic decomposition vessel does not include nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts sufficient to maintain the desired low oxygen and non-nitrifying environment for the consumption of organic acids.

[0265] In some embodiments, the first aerobic decomposition vessel includes a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), and further includes a bacterial composition adapted to thrive in such low oxygen conditions.

[0266] In some embodiments, the second aerobic decomposition vessel includes an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria. In some embodiments, the at least one chemoautotrophic microorganism may digest an ammonia compound present in the first aerobic effluent to produce nitrate compounds. In some embodiments, the second aerobic decomposition vessel includes an environment facilitating decomposition.

[0267] In some aspects, the techniques described herein relate to a system for producing an organic fertilizer from wastewater, including: at least one anaerobic digestion vessel configured to receive an organic feed stock, such as a wastewater including an organic waste, wherein the at least one anaerobic digestion vessel provides an anaerobic environment suitable to produce an anaerobic effluent; wherein the anaerobic decomposition vessel includes an organic material, such as an organic material including a sugar, protein, lipid or cellulose,preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0; at least one aerobic oxidation vessel in fluid communication with the at least one anaerobic digestion vessel, wherein the at least one aerobic oxidation vessel is configured to receive anaerobic effluent from the at least one anaerobic digestion vessel and oxygen, and digest the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel includes at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,' and at least one aerobic nitrification vessel in fluid communication with the at least one aerobic oxidation vessel, wherein the at least one aerobic oxidation vessel is configured to receive the first aerobic effluent and oxygen, and to digest the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel includes at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira.

[0268] In some aspects, the techniques described herein relate to a system, further including a port configured to receive an acid, a base or a carbon source, wherein said port is connected to at least one of the anaerobic digestion vessel, the aerobic oxidation vessel, and / or the aerobic nitrification vessel. In some embodiments, at least one of the anaerobic decomposition vessel, the first aerobic vessel, or the second aerobic vessel includes a microorganism culture derived or obtained from worm castings and / or crop waste.

[0269] In some embodiments, the anaerobic decomposition vessel includes worm castings, and the worm castings include a composition of microorganisms capable of digesting the organic material into organic acids under basic conditions, said composition of microorganisms including bacteria known for their ability to decompose complex organic materials into simpler molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter.

[0270] In some embodiments, the anaerobic decomposition vessel includes at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens Escherichia coli, Escherichia fergusonii, Escherichia albertii; Zoogloea ramigera, Zoogloea caeni, or Zoogloea oryzae.

[0271] In some embodiments, the anaerobic decomposition vessel includes a plurality of microorganisms, wherein most of the microorganisms are carbon composing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria. In some embodiments, the first aerobic decomposition vessel includes facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter. In some embodiments, the first aerobic decomposition vessel includes obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, and Eubacterium.

[0272] In some embodiments, the first aerobic decomposition vessel does not include nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts sufficient to maintain the desired low oxygen and non-nitrifying environment for the consumption of organic acids. In some embodiments, the first aerobic decomposition vessel includes a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), such as not exceeding 1 ,5ppm. Ippm, or 0.5ppm, and further includes a bacterial composition adapted to thrive in such low oxygen conditions.

[0273] In some embodiments, the second aerobic decomposition vessel includes an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria. In some embodiments, the at least one chemoautotrophic microorganism digests an ammonia compound present in the first aerobic effluent to produce nitrate compounds. In some embodiments, the second aerobic decomposition vessel includes an environment facilitating decomposition of ammonia present in the first aerobic effluent and facilitating decomposition of nitrite compounds produced during decomposition of the ammonia.

[0274] In some embodiments, the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganism and a plurality of nitrite oxidizer organism. pH and Media Cycling Between Reaction Vessels

[0275] Cycling the pH of a bioreactor, as disclosed herein, is a strategic approach to manipulate the growth and metabolic output of microbial cultures for enhanced production of specific compounds or to achieve other desired results. By intentionally varying the pH within a reactor, certain strains of microbes are selectively stressed or favored, leading to the overproduction of metabolites or the enhancement of specific biochemical pathways. This method leverages the sensitivity of microbes to pH changes, using these shifts as a tool to direct microbial activity towards the desired outcomes.

[0276] Additionally, taking advantage of a pH differential between reactors offers a dynamic method to balance or cycle the pH in a reaction vessel. By alternating the pH environment by adding effluent between more acidic and less acidic conditions across different reactors, it is possible to create a series of microenvironments that support a diverse microbial community or optimize the conditions for strains at different phases of its growth or metabolic cycle. This cycling promotes the efficiency of the reaction by maintaining desirable growth conditions and also allows for the fine-tuning of metabolic processes to enhance product pathways or yields.

[0277] Some aspects of the disclosure are related to systems and methods for controlling or monitoring the pH of at least one of effluents, inputs, and the contents of reaction vessels. In some embodiments, systems and methods maintain the pH of a media in the second aerobic decomposition vessel at 5.8 or approximately 5.8, preferably by monitoring the pH in the second aerobic decomposition vessel and introducing the first aerobic effluent from the first aerobic decomposition vessel into the second aerobic decomposition vessel when the pH falls below a pH of 5.8 or approximately 5.8. In some embodiments, the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganisms and a plurality of nitrite oxidizer organisms.

[0278] In some aspects, the techniques described herein relate to a method, further including raising the pH of a media in the second aerobic decomposition vessel by transferring the first aerobic effluent from the first aerobic decomposition vessel into the second aerobic decomposition vessel. In some embodiments, an amount of the first aerobic effluent is added to the second aerobic decomposition vessel sufficient to raise the pH of the media in the second aerobic decomposition vessel to 6.2, from, for example, a pH of 5.5. In some embodiments, a process of digesting the first aerobic effluent in the second aerobic decomposition vessel increases the pH of a media in the second aerobic decomposition vessel. In some aspects, the techniques described herein relate to a method, further including adjusting the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel. In some embodiments, the amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel is transferred when the pH of the media in the second aerobic decomposition vessel reaches 5.5 until the pH of the media in the second aerobic decomposition vessel reaches 6.2. In some embodiments, transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel is performed multiple times so as to provide a continuous culture. For instance, in some embodiments, the pH in one or both aerobic decomposition vessels is monitored by e.g., by a sensor and upon reaching a pH of 5.5 in the second aerobic decomposition vessel, the sensor initiates the opening of a valve allowing the first aerobic effluent from the first aerobic decomposition vessel to be transferred to the second aerobic decomposition vessel and once the pH in the second aerobic decomposition vessel reaches 6.2, the sensor initiates a closing of the valve, which allows the transfer of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel. This process can be initiated multiple times so as to regulate the pH in the second aerobic decomposition vessel.

[0279] In some embodiments, the pH of the first aerobic decomposition vessel may have a value of 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, or 7.2. In some embodiments, the pH of the second aerobic decomposition vessel may have a value of 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, or 6.5.

[0280] In some embodiments, the second aerobic decomposition vessel is configured to adjust the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 by transferring an amount of the first aerobic effluent, which is basic relative to other vessels, from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0281] In some embodiments, the second aerobic decomposition vessel is configured to transfer an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel when the pH of the media in the second aerobic decomposition vessel reaches 5.5, until the pH of the media in the second aerobic decomposition vessel reaches 6.2.

[0282] In some embodiments, a pH range of 7.1 to 7.8 may be maintained. In some embodiments, this pH range may be a useful range of pH values for cultivating microorganisms or for stressing microorganisms because it aligns with the secondary equivalence point of the bicarbonate buffer system. Being near, but not at, the equivalence point allows for more linear changes in pH during the dosing process. This linearity is useful for stable pH control, as it ensures that adjustments made during dosing result in predictable and manageable changes in pH. This stability is useful for the efficiency and effectiveness of the microbial activity involved in the decomposition process. In some embodiments, the pH of the first aerobic decomposition vessel may have a value within the range of 5.2 to 7.2, including values such as 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, or7.2. Similarly, in some embodiments, the pH of the second aerobic decomposition vessel may range from 5.2 to 6.5, with specific values including 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0,6.1, 6.2, 6.3, 6.4, or 6.5. Additionally, for the stage 3 setpoint, the pH may have values of 7.1,7.2, 7.3, 7.4, 7.5, 7.6, 7.7, or 7.8. The inclusion of the 7.1 to 7.8 range as a new setpoint for stage 3 allows for more flexible and efficient pH management, improving overall process stability and performance by avoiding the buffering range where pH changes are less predictable.

[0283] In some embodiments, the second aerobic decomposition vessel is configured to transfer the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel multiple times so as to provide a continuous culture where a majority of any nitrogen species are in nitrate form; and wherein a population ofammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions.

[0284] Fig. 3 is a graphical representation of experimental pH levels in a reaction over time, where the pH cycles in a wave-like pattern. The graph has arbitrary units and also includes the rate of adding effluent as a function of time. The graph displays a recurring fluctuation between higher and lower pH levels, corresponding to the addition of effluent and subsequent nitrification processes in the reactor. The line resembling a sawtooth function indicates different operating pH levels, while the line resembling a step function indicates the amount of feeding into the reactor.

[0285] Here, the first aerobic effluent is basic and causes the pH to rise, followed by a drop due to digestion within the reactor. Not shown are the levels of nitrate to ammonia levels, but the cyclical pH levels provide a proxy for determining if the goal of competition is reached. The cycling pH pattern is a control strategy to maintain a continuous culture with steady proportions of AOB and NOB, optimizing the conversion of ammonium to nitrate. The figure serves as a visual confirmation that the pH is being maintained within the desired range, allowing both bacteria to coexist and perform effectively.

[0286] Systems and methods of the disclosure aim to balance the conditions for two types of bacteria, ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB), which coexist and compete within the reactor. The desirable pH range for both AOB and NOB is 6.5 - 7, but there is a deliberate strategy to cycle the pH between higher and lower levels (6.3 to 5.4) to nurture both types. The pH cycling favors the AOB when higher and NOB when lower. The graph illustrates a successful application of this cycling strategy, with the peaks and troughs representing the alternating conditions that support the growth of both bacterium types.

[0287] In some embodiments, the methods include maintaining the pH of a media in the second aerobic decomposition vessel at approximately 5.8, preferably by monitoring the pH in the second aerobic decomposition vessel and introducing the first aerobic effluent from the first aerobic decomposition vessel into the second aerobic decomposition vessel when the pH falls below a pH of 5.8.

[0288] In some embodiments, the pH of a media in the second aerobic decomposition vessel is set such that it establishes an equilibrium between a population and / oran activity of a plurality of ammonia oxidizer microorganisms and a plurality of nitrite oxidizer organisms. In some cases, it may be advantageous to operate stage 3 at higher ranges than 6.2, such as at a pH between 7.1 to 7.8.

[0289] In some embodiments, the methods include lowering the pH of a media in the second aerobic decomposition vessel by transferring the first aerobic effluent from the first aerobic decomposition vessel into the second aerobic decomposition vessel.

[0290] In some embodiments, an amount of the first aerobic effluent is added to the second aerobic decomposition vessel in an amount sufficient to raise the pH of the media in the second aerobic decomposition vessel to 6.2.

[0291] In some embodiments, a process of digesting the first aerobic effluent in the second aerobic decomposition vessel increases the pH of a media in the second aerobic decomposition vessel.

[0292] In some embodiments, the methods include adjusting the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 or 7.1 to 7.8 by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel, wherein the pH is monitored by one or more sensors or is measured manually.

[0293] In some embodiments, the amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel is transferred when the pH of the media in the second aerobic decomposition vessel reaches 5.5 until the pH of the media in the second aerobic decomposition vessel reaches 6.2.

[0294] In some embodiments, transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel is performed multiple times so as to provide a continuous culture where a majority of any nitrogen species are in nitrate form. In some embodiments, a population of ammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions. In some embodiments, the population of ammonia oxidizing bacteria and population of nitrite oxidizing bacteria coexist at a ratio of at least one of approximately 1 :1, 1:2, 1:3, 1:4, 1:5, 1 :10, 1 :20, 1 :30, or 1:50. In some embodiments, the population of nitrite oxidizing bacteria and the population of ammonia oxidizing bacteria coexist at a ratio of at least one of approximately 1: 1, 1:2, 1 :3, 1 :4, 1:5, 1: 10, 1:20, 1 :30, or 1:50.

[0295] In some embodiments, for example in a Stage 3 reactor as described above, two or more bacteria species coexist in the same space, both competing for nutrients and oxygen. Ecological principles suggest that under desirable conditions, one bacteria strain will eventually outcompete the other to a significant extent. However, as a reactor may operate efficiently with both types of species, it is desirable in some instances to nurture the weaker bacteria, in some embodiments, the nitrite oxidizing bacteria (NOB) is weaker, and the feed is controlled in such a way that allows the two bacteria types (ammonia oxidizing bacteria (AOB) and NOB) to co-exist and provides at least a larger proportion of nitrate over nitrite. In some embodiments, the first aerobic effluent is basic, with a pH of 9, causing the pH to rise as more effluent is added. However, as effluent addition ceases, the pH drops due to nitrification processes in the second aerobic tank, where ammonium loses protons, converting them into acid protons in the solution.

[0296] Accordingly, the methods of the disclosure provide for setting the pH within a range that is favorable for AOB (more competitive) but also suitable for NOB. By way of a non-limiting example, some AOB may struggle to thrive above pH 5.4, while some NOB can perform well at pH 4.3. Both bacterium types may perform desirably at pH 6.5 -8.2. In some embodiments, the systems and methods utilize a cycling pH pattern akin to a wave, whereby feeding occurs until the pH of the vessel raises to 6.3, then pauses until it drops to 5.4 before resuming. This alternating pattern initially favors both AOB and NOB, then shifts to stress AOB while promoting NOB. This strategy results in a continuous culture whereby the majority of nitrogen species are in the nitrate form, aligning with one's goal, and where the two cultures coexist at relatively steady proportions.

[0297] In certain embodiments, one may select a maximum pH of 6.2 or approximately 6.2 to ensure a relatively linear rise and drop in pH behavior. A bicarbonate buffering system may exhibit distinct buffering regions, which can interfere with the dosing scheme if not managed carefully. Bicarbonate ions (HCO3-) are naturally produced during the microbial digestion of organic matter. As microorganisms break down organic compounds, they produce carbon dioxide (CO2), which dissolves in water to form carbonic acid (H2CO3). This carbonic acid then dissociates into bicarbonate and hydrogen ions (H+), creating a natural buffering system. This buffer chemistry can cause the pH response to feeding to become nonlinear, which might be counterproductive to the cycling strategy. In some embodiments,systems, and methods may provide a continuous culture where the majority of nitrogen species are nitrate form (the goal), and the two cultures co-exist at almost steady proportions to one another. The bicarbonate buffer system is also effective in the pH range of about 7.1 to 7.8, which aligns with the secondary equivalence point of the bicarbonate buffer system, and also aligns well with the pH range conducive for most microbial digestion processes. This pH range of the buffer system prevents significant pH fluctuations that could inhibit digestion, between 7.1 and 7.8. During microbial digestion, acidic by-products are often produced, which can lower the pH and create an inhospitable environment for microorganisms. A bicarbonate buffer at more basic pH may neutralize these acids, maintaining a stable pH and promoting continuous microbial activity and efficiency in breaking down organic matter.

[0298] Issues with systems and methods according to the disclosure may be addressed by some of the methods of adjusting the pH of vessels as described herein. For example, at times the production rate from one or more of the vessels may stall, and an overall slow production rate may occur, due, for example, to a slower feeding rate due to internal conditions within the first vessel. In some cases, the slow production rate is due to slow production from a downstream vessel, where conditions in the vessel deteriorate and the desired digestion stalls. To address such issues, the pH of one or more vessels is adjusted up or down. As described herein, various vessels may have different pH levels, such that effluent from one or more vessels is added to an upstream or downstream vessel to change the pH. Alternatively, external sources of acid, base, or buffers is added to adjust the pH. In some embodiments, the pH of Stage 2 reactor is increased by 0.2 units, i.e. to more basic pH levels. This can re-stress and / or destress various microorganisms in the vessel and jumpstart production of the overall system during an error state.

[0299] In some embodiments, an issue may arise where the pH within a system does not drop below 5.3, which may indicate a deficiency of nitrite-consuming bacteria capable of functioning in acidic environments. In such cases, ammonium-consuming bacteria tend to dominate the culture and is modulated via stress. To address this imbalance, the feeding is halted, and water from a healthy reactor or compost tea can be introduced. This intervention may prompt a drop in pH below 5.3. Once indications show that nitrite is being consumed, either through improved test strip results or by observing the pH dropping below 5.3, the set point can be resumed to 5.8. Regardless of the specific issue, high levels of aeration ismaintained within the system to promote the activity of ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB). These bacteria are aerobic and have low competitive ability, meaning that insufficient oxygen levels can severely harm these cultures. Denitrification strategies involving low oxygen conditions may also be employed.Filtering Methods

[0300] In some aspects, the techniques described herein relate to a methods and systems further including a prefiltration unit, a large pore microfilter to remove particulate matter, a tubular nanofilter, and a spiral ultrafilter. In some embodiments, filtration is performed in a single step, or in multiple with different machines and filter types. In some embodiments, one may filter solid fluids and directly produce a filtrate without any prefiltration. In some, but not all embodiments, depending on preference, filtration is sequenced, where a pre-filtration step occurs, followed by fine filtration.

[0301] In some embodiments, processing mixtures and effluents will include removing suspended solids, tannins, and larger organic molecules. In some embodiments, a filter is configured to remove suspended solids, tannins, and larger organic molecules from the second aerobic effluent. Ultrafiltration and nanofiltration are useful for effectively eliminating both dead and living bacterial cells, along with a portion of the larger organic molecules. The filtrate obtained is a sanitized solution abundant in dissolved solids such as salts and small organics, which is suitable as a fertilizer for the crops. Residues from the filtration process is recirculated back to, for example the reactor referred to herein as Stage 3. This recirculation is beneficial as it enriches the culture with bacteria, avoiding an excessively rapid removal of these microorganisms.

[0302] Filtration has multiple benefits, such as removing the decomposition products of raw organic feed through the initial stages, but yielding a solution rich in absorbable soluble salts, which is detectable by an EC sensor, and with negligible biological oxygen demand. These attributes are particularly beneficial for use in irrigation waters, for example, in a system where water is recirculated near completely. In some embodiments, maintaining sanitary conditions according to the disclosure is useful to prevent the irrigation system from being clogged with solids or bacterial growth.

[0303] Significantly, by subjecting material to highly oxidizing conditions, such as in Stage 3, most organic compounds are so thoroughly oxidized that they reach a stable state, unable to be further degraded by bacteria or mild oxidizers. This stability allows for the continual introduction of hydrogen peroxide or ozone into the irrigation water, preserving sanitary conditions and warding off foodborne and plant diseases. The choice in the permeability of the ultrafiltration or nanofiltration membrane is also strategic, removing compounds responsible for water coloration, which in turn permits the use of UV sanitation. The use of hydrogen peroxide in conjunction with UV light facilitates advanced oxidation through the generation of peroxide radicals, providing a robust oxidizing environment.

[0304] The systems and methods described herein include an advanced filtration system incorporating a prefiltration unit, a microfilter with large pores for particulate matter removal, followed by a spiral ultrafilter, and concluding with tubular nanofilter. This configuration is designed to efficiently eliminate suspended solids, tannins, and larger organic molecules from the effluent produced after the second stage of aerobic digestion. Such a comprehensive filtration sequence ensures a high-quality effluent, optimized for agricultural or other uses by removing undesirable components.

[0305] Furthermore, the process may include the recycling of the retentate, a concentrated by-product from the ultrafiltration stage, back into either the first or the second aerobic decomposition vessel. This recycling step is useful to the system by enhancing the efficiency of the decomposition process and ensuring that nutrients are not wasted but instead are reused within the system, or by treating other reaction vessels to control for, for example, pH, bacterial strain populations or ORP. Additionally, the system includes a side stream unit, which plays a role in recycling a portion of the fluids, thereby optimizing the overall water and nutrient management within the system.

[0306] In some embodiments, the methods may include a filtration sequence beginning with a prefiltration process that utilizes a large pore microfilter. This initial step is useful for removing particulate matter, which is then followed by further purification through a tubular nanofilter and / or a spiral ultrafilter. Tubular filters are typically standalone and do not require pre-filtration, whereas spiral filters might use microfiltration beforehand, with tubular microfiltration being a common choice. This sequential approach may remove suspended solids, tannins, and larger organic molecules, ensuring that the effluent from thesecond stage of aerobic digestion is purified and compatible with later treatment steps or for direct injection into an irrigation system.

[0307] Finally, the approach includes a mechanism for managing the effluent postfiltration. A portion of the filtered effluent is recycled during the filtration process, enhancing the system's sustainability. Additionally, the discharge process removes a fraction of the accumulated soluble salts, ensuring that the final effluent quality has a high concentration. In some embodiments, a filtration strategy incorporating both ultrafiltration and nanofiltration in a sequential manner efficiently produces a quality effluent, which as described herein is suitable for ultraviolet light (UV) exposure or oxidizer exposure.

[0308] In some embodiments, a retentate from the ultrafiltration process is returned to at least one of the first aerobic decomposition vessel or the second aerobic decomposition vessel. In some aspects, the techniques described herein relate to a system, further including a side stream unit configured to recycle a portion of fluids.

[0309] In some embodiments, the filtration further comprises a prefiltration process using a large pore microfilter to remove particulate matter, followed by a tubular nanofilter, and subsequently a spiral ultrafilter. "Large" in the realm of microfilters generally refers to pore sizes on the higher end of the scale for microfiltration, which typically ranges from 0.1 to 10 micrometers (pm). In some embodiments, ultrafiltration membranes can follow these initial filtration steps, which typically have pore sizes that correspond to molecular weight cut-offs (MWCO) ranging from around 1,000 to 100,000 Daltons. Nanofiltration and reverse osmosis (RO) membranes have smaller effective pore sizes, not measured in micrometers but instead in terms of their ability to reject small molecules and ions.

[0310] In some embodiments, the sequence of filtration removes suspended solids, tannins, and larger organic molecules from the second aerobic effluent. In some embodiments, a retentate from the ultrafiltration process is returned to at least one of the first aerobic decomposition vessel and the second aerobic decomposition vessel. In some embodiments, a portion of a filtered effluent is recycled during the filtration. In some embodiments, a discharge of a filtered effluent is performed, wherein said discharge removes a portion of the accumulated soluble salts. In some embodiments, the filtration comprises a sequential filtration process, first employing an ultrafiltration and then employing a nanofiltration, preferably in a series, whereinthe nanofiltration follows the ultrafiltration. Submerged membrane bioreactor configurations could be used, where the membranes are submerged directly in the reactor.

[0311] An aspect of the disclosure is also directed to the progression of effluent through a series of reaction chambers, which may play a useful role in the optimization of wastewater treatment processes. By directing the effluent from one chamber into subsequent ones, it is possible to achieve a controlled wastewater treatment, as each chamber can be tailored to remove specific unwanted reagents or to promote certain biochemical reactions. This sequential processing not only enhances the overall efficacy of the system but also allows for the strategic reuse of effluents in different stages of the wastewater treatment cycle. The variability in the composition of effluents from one, two, three, or more chambers provide a useful opportunity to design a system that can adapt to diverse wastewater characteristics.

[0312] Moreover, the analysis of effluent from individual reaction chambers offers a method for characterizing the processes occurring within a vessel, serving as a diagnostic tool or as a distinctive characteristic of a system. Understanding these processes enables the fine-tuning of operational parameters to adapt the performance of the system towards different feedstocks.

[0313] In some instances, the effluent itself, after undergoing wastewater treatment according to the disclosure, provides an effective fertilizer or source of a fertilizer, offering an environmentally friendly solution for agricultural applications. In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1. In some embodiments, before or after filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3:1. In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1. In some embodiments, the anaerobic effluent includes carbon decomposition products, such as organic acids. Nonlimiting examples of organic acids commonly found in anaerobic effluent include acetic acid, propionic acid, butyric acid, and / or lactic acid. These acids are the result of the fermentation and breakdown of organic matter by anaerobic microorganisms. Acetic acid, for instance, is an intermediate in the anaerobic digestion process and can serve as a substrate for methanogenesis, leading to the production of biogas — a mixture of methane and carbon dioxide. Similarly, propionic and butyric acids are useful intermediates that can be furtherconverted into acetic acid and hydrogen, feeding into the methanogenic phase of anaerobic digestion.

[0314] In some embodiments, the first aerobic effluent has a reduced chemical oxygen demand as compared to the anaerobic effluent. Aerobic digestion involves the use of oxygen by microorganisms to decompose organic matter, a step that may lower the COD by converting these organics into carbon dioxide, water, and biomass. The reduction in COD may indicate the treatment process's success, demonstrating a decrease in the amount of organic material present in the effluent.

[0315] For instance, after passing through an anaerobic reactor, wherein organic matter is metabolized into component compounds such as volatile fatty acids, the effluent still contains organic materials that contribute to its COD. When this effluent is introduced into an aerobic reactor, the aerobic microorganisms utilize the remaining organic compounds as their food source, further reducing the COD. This is particularly effective for wastewater containing high levels of organic pollutants that are difficult to degrade anaerobically. In some embodiments, systems and methods according to the disclosure may include adding at least one of a peroxide, humic acid and / or fulvic acid to the second aerobic effluent.

[0316] In some embodiments, the first aerobic effluent is substantially free of oxyanions of nitrogen. In some embodiments, the second aerobic effluent includes nitrites. In some embodiments, the second aerobic effluent includes reduced amounts of ammonia and / or nitrates compared to the first aerobic effluent.

[0317] In some embodiments, the ORP is measured using an electrochemical sensor such as a platinum measurement electrode or a silver / silver chloride reference electrode. In some embodiments, the ORP is measured using a colorimetric testing module configured to utilize redox indicators, which can be used to estimate the ORP of a media in a digestion vessel.Systems for processing wastewater or liquified agricultural waste

[0318] An aspect of the disclosure is also related to a system for producing an organic fertilizer from wastewater. In some embodiments, the system includes at least one anaerobic digestion vessel configured to receive an organic feed stock, such as a wastewater comprising an organic waste, wherein the at least one anaerobic digestion vessel provides ananaerobic environment suitable to produce an anaerobic effluent and, wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or agricultural crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0. In some embodiments, the system includes at least one aerobic oxidation vessel in fluid communication with the at least one anaerobic digestion vessel.

[0319] In some embodiments, the at least one aerobic oxidation vessel is configured to receive anaerobic effluent from the at least one anaerobic digestion vessel and oxygen, and digest the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent. In some embodiments, the system includes at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,'

[0320] In some embodiments, the system includes at least one aerobic nitrification vessel in fluid communication with the at least one aerobic oxidation vessel, wherein the at least one aerobic oxidation vessel is configured to receive the first aerobic effluent and oxygen, and to digest the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8. In some embodiments, the system includes at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira.

[0321] In some embodiments, the system includes a port configured to receive an acid, a base or a carbon source, wherein said port is connected to at least one of the anaerobic digestion vessel, the aerobic oxidation vessel, and / or the aerobic nitrification vessel. In some embodiments, at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from worm castings and / or crop waste e.g., a liquid culture of worm castings and / or agricultural waste can be used as an inoculum to start such a microorganism culture.

[0322] Fig. 7 shows a schematic diagram of a reactor system 700 for processing a raw input 311 with different stages indicated by various pipes illustrating a sequential processflow. Pipes 712 are connected to the Stage 1 reactor 710, where the pipes 712 indicate the flow of fluid, which is the output from the first stage of the process. Pipes 722 represent Stage 2 fluid, showing the transfer of the intermediate effluent from Stage 2 reactor 720 to the stage 3 reactor 730. Pipes for Stage 3 fluid 732, which would include effluent from the final stage of the process and the output to be used with an irrigation system.

[0323] In some embodiments, the anaerobic decomposition vessel comprises worm castings or a liquid comprising suspended worm castings or worm tea, and the worm castings, liquid comprising suspended worm castings or worm tea comprise a composition of microorganisms capable of digesting the organic material into organic acids under basic conditions, said composition of microorganisms comprising bacteria suitable for decomposing complex organic materials and mixtures into component molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter.

[0324] In some embodiments, the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens Escherichia coli, Escherichia fergusonii, Escherichia albertii; Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae.

[0325] In some embodiments, the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon composing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria.

[0326] In some embodiments, the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter.

[0327] In some embodiments, the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, and Eubacterium.

[0328] In some embodiments, the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts sufficient to maintain the desired low oxygen and non-nitrifying environment for the consumption of organic acids.

[0329] In some embodiments, the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), such as not exceeding 1.5ppm, l.Oppm, or 0.5ppm, and further comprises a bacterial population adapted to thrive in such low oxygen conditions.

[0330] In some embodiments, the second aerobic decomposition vessel comprises an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria. In some embodiments, the at least one chemoautotrophic microorganism digests an ammonia compound present in the first aerobic effluent to produce nitrate compounds.

[0331] In some embodiments, the second aerobic decomposition vessel comprises an environment facilitating decomposition of ammonia present in the first aerobic effluent and facilitating decomposition of nitrite compounds produced during decomposition of the ammonia. In some embodiments, the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganism and a plurality of nitrite oxidizer organism.

[0332] In some embodiments, the pH of a media in the second aerobic decomposition vessel is lowered by transferring the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel. In some embodiments, the second aerobic decomposition vessel is configured to digest the first aerobic effluent in the second aerobic decomposition vessel and increase the pH of a media in the second aerobic decomposition vessel. In some embodiments, the second aerobic decomposition vessel is configured to adjust the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 or 6.2to 5.5 by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0333] In some embodiments, the second aerobic decomposition vessel is configured to transfer an amount of the first aerobic effluent from the first aerobicdecomposition vessel to the second aerobic decomposition vessel when the pH of the media in the second aerobic decomposition vessel reaches 5.5, until the pH of the media in the second aerobic decomposition vessel reaches 6.2.

[0334] In some embodiments, the second aerobic decomposition vessel is configured to transfer the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel multiple times so as to provide a continuous culture, wherein a majority of any nitrogen species are in nitrate form; and wherein a population of ammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions. In some embodiments, the pH of the first and / or second aerobic decomposition vessel is evaluated or monitored such as with one or more sensors that detect pH, which facilitates the transfer of first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel when the pH threshold of 5.5 in the second aerobic decomposition vessel is reached. In some embodiments, a conduit, which connects the first aerobic decomposition vessel to a second aerobic decomposition vessel comprises a valve, which opens and closes in response to signal generated from one or more pH sensors such as when the pH in the second aerobic decomposition vessel reaches 5.5, the sensor provides a signal affecting the opening of the valve allowing an amount of the first aerobic effluent from the first aerobic decomposition vessel to enter the second aerobic decomposition vessel and when the pH of the media in the second aerobic decomposition vessel reaches 6.2, a sensor, which is the same as the sensor triggering the opening of the valve, sends a signal affecting the closing of the valve thereby stopping the flow of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0335] The system may include a prefiltration unit, a large pore microfilter to remove particulate matter, a tubular nanofilter, and a spiral ultrafilter. In some embodiments, the filter is configured to remove suspended solids, tannins, and larger organic molecules from the second aerobic effluent. In some embodiments, a retentate from the ultrafiltration process is returned to at least one of the first aerobic decomposition vessel or the second aerobic decomposition vessel. In some embodiments, the system may include a side stream unit configured to recycle a portion of the filtered water during the filtration step.

[0336] In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3:1. In some embodiments, before filtering,the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1. In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3 : 1. In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3:1. In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1. In some embodiments, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1. In some embodiments, the anaerobic effluent comprise carbon decomposition products, such as organic acids. In some embodiments, the first aerobic effluent has a reduced chemical oxygen demand as compared to the anaerobic effluent. In some embodiments, the first aerobic effluent is substantially free of oxyanions of nitrogen. In some embodiments, the second aerobic effluent comprises nitrites. In some embodiments, the second aerobic effluent comprises reduced amounts of ammonia and nitrates compared to the first aerobic effluent. In some embodiments, the ORP is measured using an electrochemical sensor comprising a platinum measurement electrode and a silver / silver chloride reference electrode. In some embodiments, the ORP is measured using a colorimetric testing module configured to use redox indicators for estimating the ORP of a media in a digestion vessel.

[0337] In some embodiments, effluents from bioreactors are characterized by their low levels of one or more contaminants, including biological oxygen demand (BOD), chemical oxygen demand (COD), nitrogenous compounds such as ammonia, nitrites, and nitrates, phosphorus, heavy metals, and / or pathogens. In some embodiments, the effluent retains high levels of nutrients, particularly nitrogen and phosphorus, which are beneficial for agricultural reuse. The nutrients should be in forms that plants can readily absorb, yet their concentration should be managed to prevent environmental issues like eutrophication. In some embodiments, the effluent contains a low concentration of suspended solids, reducing the risk of clogging and sedimentation in water bodies or irrigation systems. This typically results from effective solid-liquid separation techniques employed within the bioreactor system.

[0338] In some embodiments, effluents are free from harmful pathogens, making them safe for reuse in agriculture or aquaculture and avoiding health risks to humans and animals. Achieving this often requires UV irradiation, chlorination, or advanced filtration. In some embodiments, filtration may proceed without being overwhelmed by detritus, which allows for easier disinfection. Also, by removing certain organic acids and otherintermediaries’ other disinfection agents is compatible, such as peroxides. In some embodiments, the effluent is non-toxic to the environment, ensuring it does not harm plants, animals, or microorganisms in its discharge environment. This would entail the removal or neutralization of potential toxins during the treatment process.

[0339] In some embodiments, the effluent is suitable for reuse in various applications, including agriculture, industry, and aquaculture. This suitability often depends on the presence of beneficial constituents and the absence of harmful ones, aligning with sustainable water management and circular economy principles. In some embodiments, effluents are clear and odorless, making them more acceptable for use and easier to manage or further treat. Meeting these desirable features involves a treatment approach within the bioreactor, integrating aerobic and anaerobic digestion, nitrification-denitrification processes, chemical treatments, and advanced filtration techniques, tailored to the specific reuse requirements and local environmental regulations.Ammonium Enriched Modified Production Methods

[0340] An aspect of the disclosure is directed to compositions, and methods and systems for making such compositions, wherein the composition is an enriched ammonium solution. In some embodiments, the solution is an ammonium-only solution. Such solutions are useful for applications in wine production as a component of YAN (Yeast Assimilable Nitrogen). In general, an ammonium rich effluent is produced if aerobic digestion, such as, for example, a Stage 2 reactor (polishing of the direct anaerobic effluent) is used to reduce BOD but not further process the ammonium into nitrite / nitrate. In some embodiments, this process is controlled by adding acid to the digestor to force the sufficiently pH low to prevent or reduce nitrification and, optionally, to keep the ammonium non-volatile (which ammonia is at certain pH ranges). In some embodiments, little to no nitrification will occur below pH 5.3, but BOD reduction will continue. Desirably, citric or acetic acid is used. They are both organic acids and are expected to be digested by the same bacteria that are consuming the organic acids from the anaerobic effluent. The filtered product is preferably free of these acids / acid bases and the BOD removed. The acetic and citric acid are sources of BOD as their acid bases are simple carbon sources (acetate / citrate).

[0341] In some embodiments, the anaerobic process is operated as acidogenic to prevent excessive use of acid in, for example, aerobic polishing stages, and thereby the anaerobic processes is prevented from becoming methanogenic. As disclosed above, while methanogenic processes might be preferred for some other methods of the disclosure, notably in cases designed for producing ammonia dominated effluents, acidogenic processes is preferred. When acidogenic, the pH of the anaerobic effluent is low, such as already in the low pH of 4-5. Accordingly, in some embodiments, the effluent will be high in ammonium concentration with a low pH low, which are useful properties for the effluent for passing to an aerobic polishing step.

[0342] In some embodiments, a method of producing an ammonium-based biofertilizer from wastewater or liquified agricultural waste may include one or more of the following. Some embodiments comprise introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent. Some embodiments comprise transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel.

[0343] Some embodiments comprise digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel includes at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea. Some embodiments comprise filtering and dewatering the first aerobic effluent and producing a concentrated filtrate.

[0344] In some embodiments, the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic state at a pH from 6.0 to 9.0. In some embodiments, the anaerobic decomposition process is at least one of substantially acetogenic and substantially methanogenic. In some embodiment, the anaerobic decomposition process is at least one of substantially acetogenic, substantially hydrolytic, and substantially methanogenic. In certainembodiments, the anaerobic digestion system may include a hydrolytic digestion stage configured to facilitate the breakdown of complex organic substrates into simpler, soluble components. This hydrolytic digestion may precede subsequent stages such as acidogenesis, and can be carried out in a distinct reactor vessel or compartment adapted to support hydrolytic microbial activity. The design and operation of such a vessel may be tailored to provide environmental conditions suitable for enzymatic hydrolysis, including but not limited to parameters such as temperature, pH, retention time, and mixing regime.

[0345] The hydrolytic stage may function to depolymerize macromolecules such as polysaccharides, proteins, and lipids into their respective monomers or oligomers (e.g., sugars, amino acids, fatty acids), which may be more readily metabolized in downstream fermentation steps. Segregating the hydrolysis stage from acidogenesis may, in some implementations, enhance overall process stability and allow for improved control over intermediate compound formation. In systems configured to process substrates containing high levels of structurally complex organic material, such as lignocellulosic biomass or proteinaceous waste, inclusion of a discrete hydrolytic phase may support increased solubilization efficiency and subsequent bioconversion yield.

[0346] In some embodiments, at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel includes a microorganism culture derived from worm castings and / or crop waste, such as from liquified agricultural waste, liquified worm casting, or worm tea.

[0347] In some embodiments, the anaerobic decomposition vessel comprises worm castings, and the worm castings comprise a population of microorganisms suitable for digesting organic material into organic acids under basic conditions, said population of microorganisms comprising bacteria known for their ability to decompose complex organic materials into component molecules, such as organic acids. Preferably, bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter are utilized. In some embodiments, the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Escherichia coli, Escherichia fergusonii, and Escherichia albertii, Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae.

[0348] In some embodiments, the anaerobic decomposition vessel includes a plurality of microorganisms, wherein the majority of the microorganisms are carbon composing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria. In some embodiments, the first aerobic decomposition vessel includes facultative anaerobic bacteria suitable for consuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter.

[0349] In some embodiments, the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, or Eubacterium. In some embodiments, the first aerobic decomposition vessel does not include nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts sufficient to maintain the desired low oxygen and non-nitrifying environment for the consumption of organic acids.

[0350] In some embodiments, the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), such as not exceeding 1.5ppm, 1 ppm, or 0.5ppm or less, and further includes a bacterial composition adapted to thrive in such low oxygen conditions.

[0351] In some embodiments, the method is operated continuously. In some embodiments, the method is operated in batches. In some embodiments, the biofertilizer is compatible as a component of YAN (Yeast Assimilable Nitrogen).Compositions

[0352] An aspect of the disclosure is directed to the production of an ammonium- only solution. This solution may be useful for applications in wine production as a component of YAN (Yeast Assimilable Nitrogen). Traditional activated sludge systems fall short in producing highly concentrated digestate due to their slow processing and high energy demands. In contrast, a multistage-reactor approach according to the disclosure may enhance efficiency and maximize digestate concentration beyond what is achievable with aerobic digestion alone. Anaerobic digestion as the first stage, can be markedly efficient, consuming only 5% of the carbon and nitrogen in the feed, compared to the 35% typically utilized in aerobic digestion.This process not only facilitates high biogas production but, when operated under acidic conditions, can halt methane production, allowing subsequent stages to refine organic acids and produce bicarbonate for nitrification. Achievable ammonium concentrations can reach up to 3000 ppm in anaerobic reactors, surpassing the 500 ppm limit of aerobic-activated sludge processes. This method also benefits from a reduced footprint compared to traditional aerobic systems, leading to fewer requirements for filtration and dewatering equipment.

[0353] Furthermore, to address regulations on the sale of organic fertilizers with high nitrogen content, which depend on the carbon-to-nitrogen ratio, this disclosure provides for solutions compatible with the addition of mined humic or fulvic acid compositions. These compounds are stable, highly soluble, and resistant to decomposition even in the presence of hydrogen peroxide, unlike ozone, and do not promote microbial growth. By incorporating humic acids, the carbon-to-nitrogen ratio of the effluent is increased above the 3: 1 threshold, allowing for the unrestricted use of the fertilizer in irrigation systems treated with peroxide, without risking microbial proliferation. Additionally, the use of organic humic or fulvic acids brings further benefits to crops. In some embodiments, a liquid fertilizer produced can be characterized by a COD of less than 125 mg / L and a BOD of less than 30 mg / L. In some embodiments, the solution may remain stable when peroxides are introduced and so the solution may include organic acids such as humic acid, fulvic acid, citric acid, or acetic acid. In some embodiments, this composition is produced by processing wastewater through an anaerobic vessel followed by sequential aerobic vessels. In some embodiments, the process is controlled by maintaining specific pH and ORP conditions conducive to the growth of specific organic acid- consuming and chemoautotrophic microorganisms.

[0354] An aspect of the disclosure relates to a liquid fertilizer produced from organic waste. In some embodiments, the liquid fertilizer has a chemical oxygen demand (COD) of less than 125 mg / L, and a biological oxygen demand (BOD) of less than 30 mg / L. In some embodiments, the liquid fertilizer is stable upon addition of peroxides. In some embodiments, the liquid fertilizer includes an organic acid. In some embodiments, the liquid fertilizer includes an organic acid selected from the group consisting of humic acid, fulvic acid, citric acid, and acetic acid. In some embodiments, the composition is solid or liquid and is produced by introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent.

[0355] In some embodiments, a system for producing the liquid fertilizer includes an anaerobic decomposition vessel which may contain an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, In some embodiments, the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0;

[0356] In some embodiments, the system includes a capability of transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel.

[0357] In some embodiments, the system includes a capability of digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between - 1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,'

[0358] In some embodiments, the system includes a capability of transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel. In some embodiments, a conduit, which connects the first aerobic decomposition vessel to a second aerobic decomposition vessel comprises a valve, which opens and closes in response to signal generated from one or more pH sensors such as when the pH in the second aerobic decomposition vessel reaches 5.5, the sensor provides a signal affecting the opening of the valve allowing an amount of the first aerobic effluent from the first aerobic decomposition vessel to enter the second aerobic decomposition vessel and when the pH of the media in the second aerobic decomposition vessel reaches 6.2, a sensor, which can be the same as the sensor triggering the opening of the valve, sends a signal affecting the closing of the valve thereby stopping the flow of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

[0359] In some embodiments, the system includes a capability of digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or anoxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira, e.g., a bacterial population obtained from earthworm castings, liquified agricultural waste, or worm tea.

[0360] Base media contactor

[0361] In some embodiments, a base media contactor may apply specifically to the second aerobic decomposition vessel or any aerobic treatment unit positioned downstream of stage 1 as described herein. For example, a base media contactor may be positioned downstream from, or integrated with, a stabilization or maturation tank. These embodiments supports and enhances the function of the aerobic treatment stages by modulating both the chemical and physical characteristics of the process stream prior to or during biological oxidation. Implementing a base media contactor in this configuration serves functions as chemical conditioning. In some embodiments, the base media contactor may assist with pH stabilization and / or ion supplementation with the intention of supporting microbial health, and the quality of the final biofertilizer output.

[0362] In some embodiments, the base media contactor is fluidically integrated with an aerobic oxidation vessel — such as the second aerobic decomposition unit (Stage 2) — to enable controlled interaction between the process fluid and alkaline-reactive solids. This interaction can elevate pH through the dissolution of carbonate or oxide components, and may optionally simultaneously introduce beneficial mineral ions, such as calcium, magnesium, or phosphate, into the treatment stream. The system may operate in either an upflow or downflow configuration depending on, for example, the desired balance of residence time, ion release rate, and physical filtration. The contactor may also serve as a preliminary source of filtration in a downflow operation, reducing particulate loading prior to finer filtration units. This embodiment aligns with the disclosure as set forth in alternatives 34, 39, and 57-61 by contributing to pH control, filtration, and / or mineral enhancement.

[0363] In some embodiments, at least one base media contactor is fluidically connected to at least one aerobic oxidation vessel — such as the second aerobic decomposition vessel — to modulate pH and supplement mineral ions beneficial to microbial activity andbiofertilizer quality. The contactor comprises a bed of base media, which may include, but is not limited to, calcite, dolomitic limestone, phosphate rock, Corosex™, or other alkaline- reactive solids. Process fluid may flow through the media bed in either an upflow or downflow configuration. In the upflow arrangement, fluid residence time is controlled to allow dissolution of alkaline constituents, particularly carbonates and oxides, resulting in elevated pH. In the downflow configuration, the packed media bed may additionally function as a physical filtration medium, effectively reducing suspended solids and particulate loading prior to downstream filtration stages, such as microfilters (with pore sizes typically between 1 and 10 micrometers), spiral ultrafilters, or tubular nanofilters.

[0364] In some embodiments, when the base media contactor is in fluidic communication with the aerobic oxidation vessel, dissolution of the carbonate or oxide-based components raises solution pH by releasing alkaline anions (e.g., carbonate, bicarbonate, or hydroxide ions). Associated mineral counterions (e.g., calcium, magnesium, and / or phosphate ions) are introduced into the solution and may provide agronomic benefits by enhancing the nutrient profile of the resulting biofertilizer product using sources compatible with organic production standards. This controlled ion addition complements broader pH management strategies, such as co-dosing aqueous ammonia in concert with Stage 2 effluent dosing as a medium during aerobic nitrification, as further described herein.

[0365] Control Regime and Hydraulic Retention Management

[0366] In various embodiments, the system and methods disclosed herein incorporate an integrated control regime for managing hydraulic retention, solids loading, and operational stability across multiple biological treatment stages. This control approach is particularly relevant in systems comprising at least one anaerobic decomposition vessel and at least two aerobic decomposition vessels arranged in sequence. The disclosed regime supports stable, continuous operation by dynamically coordinating process conditions across these stages in response to real-time monitoring data.

[0367] Unlike conventional batch or semi-continuous systems, which rely on fixed time intervals or isolated feedback loops, the present system is designed to respond to biochemical indicators and operational parameters in a coordinated and adaptive manner. This continuous and interdependent operation offers improved microbial health, greater system resilience, and more consistent production of biofertilizer products.

[0368] The control strategies described herein may be implemented in conjunction with any of the other system components or operational features disclosed throughout this application, including but not limited to base media contactors for pH modulation and mineral ion supplementation, multi-stage filtration systems, pH cycling protocols, and / or nutrient dosing schedules. The coordination of retention times, aeration rates, and chemical inputs between the anaerobic and aerobic stages enables the system to maintain a stable biochemical environment that supports efficient nitrification, organic acid digestion, and ammonia oxidation, while mitigating common failure modes such as nitrite accumulation, pH collapse, or ammonium toxicity. This integrated operational logic, supported by continuous data acquisition and responsive control actions, distinguishes the system as a scalable and robust platform for high-quality biofertilizer production, including in settings where organic certification or stringent nutrient control is required.

[0369] The hydraulic and microbial control features described herein may be used in any suitable combination with the previously disclosed features and embodiments, including the use of alkaline mineral media, ORP-based aeration regulation, pH-buffered dosing from Stage 2 to Stage 3, and effluent conditioning via base media contactors. The design flexibility afforded by this framework allows for the adjustment of process dynamics in response to operational challenges, such as changes in feedstock composition, seasonal temperature variation, or upstream imbalances. Moreover, the use of real-time measurements — including FOS / TAC ratios, electrical conductivity, bicarbonate availability, nitrite concentration, and pH trends — enables a closed-loop response to evolving process conditions, enhancing the resilience and performance of the treatment system under both steady-state and transitional modes of operation.

[0370] Accordingly, the features of the control regime described herein are intended to be combinable with other elements of the invention, and written support is provided for claims that integrate these elements into a unified treatment system. The use of proportional solids loading in Stage 1, strategic aeration modulation in Stage 2, and conditional nitrification recovery protocols in Stage 3 may be deployed together or independently, in various configurations, without departing from the scope of the invention. These features are not limited to specific control software or hardware implementations and may be realized throughprogrammable logic controllers, sensor-integrated dosing systems, or manual operational protocols, depending on the needs and scale of a given installation.

[0371] In certain embodiments, the processes disclosed herein utilize a controlled hydraulic retention and solids loading regime coordinated across multiple treatment stages, including at least one anaerobic decomposition vessel (Stage 1) and at least two aerobic decomposition vessels (Stages 2 and 3). Such control schemes promote overall microbial stability, maintain optimal process conditions, and improve the consistency of biofertilizer quality.

[0372] In some embodiments, solids loading and hydraulic retention parameters in the anaerobic decomposition vessel (Stage 1) are managed proportionally relative to effluent transfer rates into subsequent aerobic decomposition vessels. By maintaining proportional loading control, the total ionic concentration, which may be monitored through electrical conductivity (EC), remains substantially constant, thereby supporting steady-state microbial performance throughout the multi-stage system.

[0373] In certain embodiments, the anaerobic decomposition vessel (Stage 1) is monitored by measuring organic acid accumulation relative to total alkalinity - FOS / TAC ratio. When the FOS / TAC ratio indicates an increase in organic acids beyond a predetermined threshold — signaling a potential acid-phase imbalance or methanogenic inhibition — the solids loading rate or hydraulic throughput into Stage 1 must be reduced. Such reduction minimizes the risk of ammonium toxicity, elevated total dissolved solids (TDS), and bicarbonate shortages, each of which may adversely affect downstream nitrification processes.

[0374] In some embodiments, effluent from Stage 1 is transferred into a first aerobic decomposition vessel (Stage 2). Stage 2 typically has a relatively short hydraulic retention time and is infrequently the rate-limiting step. Nevertheless, under certain conditions where extended retention within other coupled digesters becomes necessary, aeration within Stage 2 may be strategically reduced to minimal levels sufficient only to prevent excessive microbial death. Such reduced aeration in Stage 2 limits excessive ammonium volatilization (air stripping of ammonia). Additionally, minimizing aeration prevents or reduces unintended partial nitrification and the accumulation of nitrite, which can significantly depress the pH of Stage 2 effluent. If Stage 2 effluent pH becomes depressed due to prolonged retention and partial nitrification, subsequent dosing strategies into Stage 3 may inadvertently overshoottarget pH setpoints by introducing excessive quantities of nitrite-containing effluent, or fail altogether. Preventing or reducing nitrite formation by managing aeration and retention times in Stage 2 thus mitigates substrate toxicity, reduces risk of pH overshoot during dosing in Stage 3, and supports overall system stability.

[0375] In further embodiments, effluent from Stage 2 flows into at least a second aerobic decomposition vessel (Stage 3), wherein chemoautotrophic nitrifying microorganisms — such as those belonging to the genera Nitrosomonas, Nitrobacter, Nitrospira, and related organisms — conduct nitrification. Stage 3 operations may, at times, become sensitive to elevated nitrite concentrations resulting from upstream digester imbalances or natural culture behaviors. In such cases, one or more corrective measures may optionally be employed, including controlled large-amplitude pH cycling, intentional dilution with fresh water to mitigate nitrite substrate inhibition, and / or controlled exposure to alkaline mineral media, such as calcite, dolomitic limestone, or magnesium oxide. These measures manage inhibitory nitrite levels, thereby restoring or maintaining optimal nitrification activity.

[0376] Specifically, Stage 3 nitrification rates may be intentionally reduced through a strategic reduction in dosing frequency or by allowing larger fluctuations in pH, including periodic extensions below typical operating ranges, followed by rapid return to standard operating pH conditions. Such controlled pH cycling beneficially preserves nitrifying culture viability, avoiding prolonged lag phases and facilitating quicker system recovery upon resolution of upstream conditions. pH ranges 7-3, 6.5-4, 6-4,5.5-4, or 5.5-3.

[0377] In some embodiments, Stage 3 is preferably maintained under conditions that avoid prolonged starvation or stable pH intervals that may compromise microbial activity. For instance, Stage 3 may not remain unfed or without substrate addition for intervals exceeding approximately 18 hours or sustain stable (non-decreasing) pH levels for longer than about 4 hours. Adherence to these parameters beneficially prevents or reduces substantial microbial deterioration and prolonged lag and growth phase recovery times.

[0378] In further embodiments, an integrated closed-loop control framework dynamically coordinates operational parameters among Stage 1, Stage 2, and / or Stage 3, based on real-time monitoring data, including but not limited to FOS / TAC ratios, electrical conductivity, ammonium concentration, nitrite levels, bicarbonate availability, and pH dynamics. Such an integrated approach beneficially enables responsive and strategicmanagement of hydraulic retention times, solids loading rates, aeration levels, base media exposure, and / or nutrient availability. Consequently, this flexible control scheme improves digestor stability, prevents or reduces unintended substrate toxicities or alkalinity imbalances, and advantageously supports the consistent production of high-quality biofertilizers.

[0379] In some embodiments, the base mineral media employed in these control strategies comprises naturally occurring, mined minerals permitted within organic agricultural practices, thereby permitting the resultant biofertilizer products to meet organic certification standards and market preferences.Examples

[0380] Example 1: A benefit of the systems and methods of the disclosure is illustrated by the following comparative example. In this example, an individual decided to transition their conventional greenhouse to utilize an organic fertilizer supply. Upon discovering a drum of liquid organic fertilizer with a 2-1-2 NPK ratio, which was less potent than the conventional solution previously used, the assumption was made that simply diluting the fertilizer would suffice for the plants' nutritional consumption.

[0381] However, within days, issues emerged, including alarms signaling low flow on certain valves and workers reporting a sewage-like odor. Upon inspection, the greenhouse was filled with a foul smell, plants appeared dehydrated, and the irrigation tanks were overrun with slime, emanating a putrid odor from the fertilizer. It became evident that the fertilizer used had fouled.

[0382] The plants exhibited signs of nitrogen and iron deficiencies, prompting a food safety team to conduct water and plant sample analyses. The oversight was the direct addition of organic fertilizer to the irrigation water, not recognizing that organic fertilizers precondition microbial decomposition in order to release nutrients in a form accessible to crops. Organic fertilizers encapsulate nutritional salts within large organic molecules, which would entail microbial action to convert them into usable ions for the plants.

[0383] The existing system lacked the capability to facilitate microbial decomposition. The introduction of the fertilizer initiated a decomposition process by naturally occurring bacteria. Despite having sanitation measures such as UV lamps or oxidizers, the system inadvertently turned into a breeding ground for decomposition. Without an adequateaeration system, the microbial activity swiftly depleted oxygen levels, leading to an anaerobic environment and the proliferation of anaerobic bacteria, further exacerbating the breakdown process. This sequence of events not only filled the facility with foul odors but also posed significant food safety risks and deprived plant roots of oxygen, hindering their ability to absorb the incompletely decomposed organic fertilizer, thereby leading to nutritional deficiencies.

[0384] This example underscores the importance of properly decomposing and composting organic fertilizers before their application to ensure they are in a form that plants can effectively utilize, highlighting the potential drawbacks of direct incorporation into irrigation systems.Example 2

[0385] In the operation of anaerobic digesters, two distinct conditions can be observed: acidic or methanogenic. When operated under acidic conditions, the environment is predominantly conducive to acid-forming bacteria. These bacteria decompose complex feedstocks into smaller, volatile organic acids such as butyric acid, reminiscent of the smell of rotten eggs, and acetic acid, which has a vinegar-like odor. The downside of acidic operation in an anaerobic digester is not just the unpleasant smell; it also indicates that the digestion process is not fully efficient, leaving valuable biogas components unproduced.

[0386] On the other hand, when digesters operate under methanogenic conditions, another group of bacteria, known as methanogens, thrive. These microorganisms consume the organic acids produced in the earlier stage of digestion, converting them into methane and carbon dioxide, which are the desired end products in biogas applications. Methanogens enhance the yield of methane, but challenges arise in continuous flow cultures of anaerobes. Despite the activity of methanogens, the effluent extracted from the reactor contains amounts of undecomposed organic acids. The concentration of these acids in the effluent can vary, reflecting the effectiveness of methanogen maintenance within the reactor. When methanogens are not adequately supported — due to factors such as improper pH balance, temperature fluctuations, or nutrient imbalances — the digestion process skews towards acidic operation. This imbalance can lead to an increase in the concentration of unpleasant-smelling organicacids in the effluent, reduced methane yield, and overall inefficiency of the biogas production process.Example 3

[0387] In contrast to the problematic scenarios previously discussed, consider a situation where a greenhouse operator adopts a multistage-reactor approach for organic fertilizer production and application, adhering to the methodologies outlined in a specific disclosure. This approach employs a series of reactors in sequence, significantly improving efficiency and allowing for the maximization of digestate concentration, a feat unattainable with conventional aerobic digestion methods.

[0388] Beginning with anaerobic digestion as the first stage, this method proves to be highly efficient, consuming merely 5% of the carbon and nitrogen present in the feedstock, in stark contrast to the 35% consumption rate typical of aerobic processes. This efficiency leap not only optimizes resource use but also enhances the overall sustainability of the fertilizer production process.

[0389] Following the anaerobic stage, subsequent aerobic digestion steps further refine and stabilize the digestate, ensuring that it is in a form readily assimilable by plants. This multistage processing effectively addresses the limitations seen in direct organic fertilizer application scenarios. The resulting product is a highly concentrated, nutrient-rich digestate, free of the drawbacks associated with incomplete decomposition and microbial imbalance.

[0390] Employing this approach, the greenhouse avoids the issues of oxygen depletion, unpleasant odors, and nutritional deficiencies. Instead, it benefits from an optimized fertilizer that supports plant health and productivity without compromising the system's integrity or safety. This example illustrates how following a process according to the disclosure can avoid potential pitfalls associated with liquid organic fertilization for agriculture.Example 4: Demonstration of Nitrogen Species Stabilization in Stage 3 Bioreactors

[0391] Two Stage 3 reactor samples were analyzed to demonstrate nitrogen speciation outcomes under different operating conditions. These samples were obtained from a bioreactor system processing decomposed agricultural waste through a multi-stage microbial treatment method. The results illustrate the formation of nitrate as the dominant nitrogenspecies under both active and passive reactor management conditions, with variable impacts on pH and residual nitrite clearance.

[0392] Sample Stage3R4 was allowed to stabilize with no active feed input, representing a paused batch mode operation. Stage3R4 was a demonstration batch that underwent a pause in feed input, allowing the material to stabilize naturally without active dosing. This approach led to a significant drop in pH to 2.22. The sample exhibited dominant nitrate nitrogen levels at 597 mg / L, with nitrite nitrogen at only 1.56 mg / L and residual ammoniacal nitrogen at 69 mg / L. This result indicates that passive acidification from microbial processes effectively cleared residual nitrite, a less stable intermediate, and allowed nitrate to accumulate as the primary nitrogen species.

[0393] Sample Stage3R9 was operated at a controlled pH setpoint under continuous feed conditions. This represents standard operating parameters for steady-state production. The nitrogen speciation profile again showed nitrate dominance with low nitrite and reduced ammonium concentrations.

[0394] Table 3: Nitrogen Speciation and pH Results from Stage 3 Reactor Effluent Samples (Paused vs. Normal Operation)Ammoniacal Nitrogen Nitrate-Nitrogen Nitrite-Nitrogen Sample ID pH(mg / L) (mg / L) (mg / L)Stage3R4 2.22 69 597 1.56Stage3R9 6.07 58 461 0.47

[0395] These results show the effectiveness of the disclosed methods in producing a nitrate-rich effluent under both controlled and free-acidifying conditions. They also give evidence to support the operational flexibility of the system, allowing for active or passive nitrite clearance as needed. This flexibility can be leveraged to optimize production cycles, manage feed interruptions, or enhance stability in storage.

Claims

WHAT IS CLAIMED IS:

1. A method of producing an organic fertilizer from a wastewater, comprising: introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent; wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained in an anaerobic condition at a pH from 6.0 to 9.0; transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel; digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent, wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,' transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel; digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira,' and filtering at least the first or second aerobic effluent or both;wherein the second aerobic effluent comprises inorganic nitrogen in oxyanion form and has a carbon to nitrogen ratio greater than or equal to 3: 1.

2. The method of claim 1 , wherein at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from an inoculum.

3. The method of claim 2, wherein the anaerobic decomposition vessel comprises an inoculum selected from a group consisting of compost, liquid compost worm castings, crop waste, microbial inoculum, and commercial inoculum, wherein the inoculum comprises a composition of microorganisms capable of digesting the organic material into at least organic acids and ammonium (but also include methane in here as additional scope) under basic conditions, said composition of microorganisms comprising bacteria known for their ability to decompose complex organic materials into simpler molecules, including organic acids, ammonium, and methane, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter. (include methanogens)4. The method according to any one of the preceding claims, wherein the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Escherichia coli, Escherichia fergusonii, Escherichia albertii, and Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae, and Methanogens including species such as Methanobacterium, Methanosarcina, Methanococcus, Methanomicrobium, Methanospirillum, Methanothermobacter, and Methanoculleus .

5. The method according to any one of the preceding claims, wherein the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon decomposing bacteria, and wherein the plurality of microorganisms does not include ammonium decomposing bacteria.

6. The method according to any one of the preceding claims, wherein the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of Lactobacillus, Streptococcus, and Enterobacter.

7. The method according to any one of the preceding claims, wherein the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, e.g., species from the genera Bacteroides, Clostridium, or Eubacterium.

8. The method according to any one of the preceding claims, wherein the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts compatible with a desired low oxygen and non-nitrifying environment for a least one of reduction of chemical and biological oxygen demand and consumption of organic acids.

9. The method according to any one of the preceding claims, wherein the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), and further comprises a bacterial composition adapted to thrive in such low oxygen conditions.

10. The method according to any one of the preceding claims, wherein the second aerobic decomposition vessel comprises an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria.

11. The method according to any one of the preceding claims, wherein the at least one chemoautotrophic microorganism digests an ammonia compound present in the first aerobic effluent to produce nitrate compounds.

12. The method according to any one of the preceding claims, wherein the second aerobic decomposition vessel comprises an environment facilitating decomposition of ammonia present in the first aerobic effluent and facilitating decomposition of nitrite compounds produced during decomposition of the ammonia.

13. The method according to any one of the preceding claims, comprising maintaining the pH of a media in the second aerobic decomposition vessel at approximately 5.8 or 7.4, preferably by monitoring the pH in the second aerobic decomposition vessel and introducing the first aerobic effluent, which is preferably basic, from the first aerobic decomposition vessel into the second aerobic decomposition vessel when the pH falls below a pH of 5.8.

14. The method according to any one of the preceding claims, wherein the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganisms and a plurality of nitrite oxidizer organisms.

15. The method according to any one of the preceding claims, further comprising raising the pH of a media in the second aerobic decomposition vessel by transferring the first aerobic effluent from the first aerobic decomposition vessel into the second aerobic decomposition vessel.

16. The method of claim 15, wherein an amount of the first aerobic effluent is added to the second aerobic decomposition vessel sufficient to raise the pH of the media in the second aerobic decomposition vessel to 6.2 or 7.6.

17. The method according to any one of the preceding claims, wherein a process of digesting the first aerobic effluent in the second aerobic decomposition vessel decreases (acidifies) the pH of a media in the second aerobic decomposition vessel.

18. The method according to any one of the preceding claims, further comprising adjusting the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 or 7.1 to 7.8 by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

19. The method of claim 18, wherein the amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel is transferred when the pH of the media in the second aerobic decomposition vessel reaches 5.5 until the pH of the media in the second aerobic decomposition vessel reaches 6.2 or when the pH reaches 7.8 until the pH of the media in the second aerobic decomposition vessel reaches 7.1.

20. The method according to any one of the preceding claims, wherein transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel is performed multiple times so as to provide a continuous culture where a majority of any nitrogen species are in nitrate form; and wherein a population of ammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions.

21. The method according to any one of the preceding claims, wherein the filtration further comprises a prefiltration process using a large pore microfilter to remove particulate matter, preferably followed a spiral ultrafilter preferably subsequently followed by a tubular nanofilter.

22. The method according to claim 21, wherein the sequence of filtration removes suspended solids, tannins, and larger organic molecules from the second aerobic effluent.

23. The method according claims 21 or 22, wherein a retentate from the filtration process is returned to at least one of the first aerobic decomposition vessel and the second aerobic decomposition vessel.

24. The method according to any one of the preceding claims, further comprising a side stream process, wherein a portion of a filtered effluent is recycled during the filtration.

25. The method according to any one of the preceding claims, wherein a discharge of a filtered effluent is performed, wherein said discharge removes a portion of the accumulated suspended solids.

26. The method according to any one of the preceding claims, wherein the filtration comprises a sequential filtration process, first employing an ultrafiltration and then employing a nanofiltration, preferably in a series, wherein the nanofiltration follows the ultrafiltration.

27. The method according to any one of the preceding claims, wherein, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3 : 1.

28. The method according to any one of the preceding claims, wherein the anaerobic effluent comprises carbon decomposition products, such as organic acids.

29. The method according to any one of the preceding claims, wherein the first aerobic effluent has a reduced biological oxygen demand as compared to the anaerobic effluent.

30. The method according to any one of the preceding claims, wherein the first aerobic effluent is substantially free of oxyanions of nitrogen.

31. The method according to any one of the preceding claims, wherein the second aerobic effluent comprises nitrates.

32. The method according to any one of the preceding claims, wherein the second aerobic effluent comprises reduced amounts of ammonia compared to the first aerobic effluent.

33. The method according to any one of the preceding claims, further comprising adjusting the pH of the anaerobic effluent, the first aerobic effluent, and / or the second aerobic effluent.

34. The method according to any one of the preceding claims, further comprising adding at least one of base and a carbon source, such as molasses, saccharides, lime, and bicarbonates, to the second aerobic decomposition vessel and / or the first aerobic decomposition vessel.

35. The method according to any one of the preceding claims, wherein the ORP is measured using an electrochemical sensor comprising a platinum measurement electrode and a silver / silver chloride reference electrode.

36. The method according to any one of the preceding claims, wherein the ORP is measured using a colorimetric testing module configured to use redox indicators for estimating the ORP of a media in a digestion vessel.

37. The method according to any one of the preceding claims, further comprising adding at least one of a peroxide, ozone, humic acid and / or fulvic acid to the second aerobic effluent or a filtrate thereof.

38. A system for producing an organic fertilizer from wastewater, comprising: at least one anaerobic digestion vessel configured to receive an organic feed stock, such as a wastewater comprising an organic waste, wherein the at least one anaerobic digestion vessel provides an anaerobic environment suitable to produce an anaerobic effluent; wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0;at least one aerobic oxidation vessel in fluid communication with the at least one anaerobic digestion vessel, wherein the at least one aerobic oxidation vessel is configured to receive anaerobic effluent from the at least one anaerobic digestion vessel and oxygen, and digest the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,' and at least one aerobic nitrification vessel in fluid communication with the at least one aerobic oxidation vessel, wherein the at least one aerobic oxidation vessel is configured to receive the first aerobic effluent and oxygen, and to digest the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira.

39. The system according claim 38, further comprising a port configured to receive an acid, a base or a carbon source, wherein said port is connected to at least one of the anaerobic digestion vessel, the aerobic oxidation vessel, and / or the aerobic nitrification vessel.

40. The system according to any one of claims 38 to 39, wherein at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from an inoculum, preferably an inoculum of compost, liquid compost, worm castings and / or crop waste.

41. The system according to any one of claims 38 to 40, wherein the anaerobic decomposition vessel comprises an inoculum comprising a composition of microorganisms capable of digesting the organic material into organic acids under basic conditions, said composition of microorganisms comprising bacteria known for theirability to decompose complex organic materials into simpler molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter.

42. The system according to any one of claims 38 to 41, wherein the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens Escherichia coli, Escherichia fergusonii, Escherichia albertir, Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae, and Methanogens includiing species such as Methanobacterium, Methanosarcina, Methanococcus, Methanomicrobium, Methanospirillum, Methanothermobacter, and Methanoculleus .

43. The system according to any one of claims 38 to 42, wherein the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon decomposing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria.

44. The system according to any one of claims 38 to 43, wherein the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable of consuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter.

45. The system according to any one of claims 38 to 44, wherein the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, and Eubacterium.

46. The system according to any one of claims 38 to 45, wherein the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts compatible with a desired low oxygen and non-nitrifying environment for at least one of reduction of chemical and biological oxygen demand and of consumption of organic acids.

47. The system according to any one of claims 38 to 46, wherein the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration notexceeding 2 mg / L (2 ppm), and further comprises a bacterial composition adapted to thrive in such low oxygen conditions.

48. The system according to any one of claims 38 to 47, wherein the second aerobic decomposition vessel comprises an environment hosting a mixture of bacteria, said mixture including both ammonia-digesting bacteria and nitrite-digesting bacteria.

49. The system according to any one of claims 38 to 48, wherein the at least one chemoautotrophic microorganism digests an ammonia compound present in the first aerobic effluent to produce nitrate compounds.

50. The system according to any one of claims 38 to 49, wherein the second aerobic decomposition vessel comprises an environment facilitating decomposition of ammonia present in the first aerobic effluent and facilitating decomposition of nitrite compounds produced during decomposition of the ammonia.

51. The system according to any one of claims 38 to 50, wherein the pH of a media in the second aerobic decomposition vessel establishes an equilibrium between a population and / or an activity of a plurality of ammonia oxidizer microorganism and a plurality of nitrite oxidizer organism.

52. The system according to any one of claims 38 to 51, wherein the pH of a media in the second aerobic decomposition vessel is raised by transferring the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

53. The system according to any one of claims 38 to 52, wherein the second aerobic decomposition vessel is configured to digest the first aerobic effluent in the second aerobic decomposition vessel and increase the pH of a media in the second aerobic decomposition vessel.

54. The system according to any one of claims 38 to 53, wherein the second aerobic decomposition vessel is configured to adjust the pH of a media in the second aerobic decomposition vessel from 5.5 to 6.2 or 7.1 to 7.8 by transferring an amount of the first aerobic effluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel.

55. The system according to any one of claims 38 to 54, wherein the second aerobic decomposition vessel is configured to transfer an amount of the first aerobiceffluent from the first aerobic decomposition vessel to the second aerobic decomposition vessel when the pH of the media in the second aerobic decomposition vessel reaches 5.5, until the pH of the media in the second aerobic decomposition vessel reaches 6.2.

56. The system according to any one of claims 38 to 55, wherein the second aerobic decomposition vessel is configured to transfer the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel multiple times so as to provide a continuous culture where a majority of any nitrogen species are in nitrate form; and wherein a population of ammonia oxidizing bacteria and a population of nitrite oxidizing bacteria coexist at steady proportions.

57. The system according to any one of claims 38 to 56, further comprising a prefiltration unit, a microfilter with a pore size between 1 and 10 micrometers to remove particulate matter, a tubular nanofilter, and a spiral ultrafilter.

58. The system according to any one of claims 38 to 57, wherein the filter is configured to remove suspended solids, tannins, and larger organic molecules from the second aerobic effluent.

59. The system according to any one of claims 38 to 58, wherein a retentate from the ultrafiltration process is returned to at least one of the first aerobic decomposition vessel or the second aerobic decomposition vessel.

60. The system according to any one of claims 38 to 59, further comprising a side stream unit configured to recycle a portion of the retentate during the filtration step.

61. The system according to any one of claims 38 to 60, wherein, before filtering, the second aerobic effluent has a carbon to nitrogen ratio greater than or equal to 3: 1, preferably through the addition of organic acids to increase the carbon to nitrogen ratio without substantially increasing the biological oxygen demand.

62. The system according to any one of claims 38 to 61, wherein the anaerobic effluent comprises carbon decomposition products, such as organic acids.

63. The system according to any one of claims 38 to 62, wherein the first aerobic effluent has a reduced chemical oxygen demand as compared to the anaerobic effluent.

64. The system according to any one of claims 38 to 63, wherein the first aerobic effluent is substantially free of oxyanions of nitrogen.

65. The system according to any one of claims 38 to 64, wherein the second aerobic effluent comprises nitrites.

66. The system according to any one of claims 38 to 65, wherein the second aerobic effluent comprises reduced amounts of ammonia compared to the first aerobic effluent.

67. The system according to any one of claims 38 to 66, wherein the ORP is measured using an electrochemical sensor comprising a platinum measurement electrode and a silver / silver chloride reference electrode.

68. The system according to any one of claims 38 to 67, wherein the ORP is measured using a colorimetric testing module configured to use redox indicators for estimating the ORP of a media in a digestion vessel.

69. A method of producing an ammonium-based biofertilizer from a wastewater, comprising: introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent; wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0; transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel; and digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea.

70. The method of claim 69, further comprising filtering and dewatering the first aerobic effluent and producing a concentrated filtrate.

71. The method according to any one of claims 69 to 70, wherein the anaerobic decomposition process is at least one of substantially acetogenic and substantially methanogenic.

72. The method according to any of one of claims 69 to 71 , wherein at least one of the anaerobic decomposition vessel, the first aerobic vessel, and the second aerobic vessel comprises a microorganism culture derived from an inoculum comprising at least one of compost, liquid compost, worm castings, crop waste, microbial inoculum, and commercial inoculum.

73. The method according to any of claims 69 to 72, wherein the anaerobic decomposition vessel comprises a composition of microorganisms capable of digesting the organic material into organic acids under basic conditions, said composition of microorganisms comprising bacteria known for their ability to decompose complex organic materials into simpler molecules, including organic acids, such as bacteria selected from the group consisting of Pseudomonas, Bacillus, and Azotobacter, and Methanogens such as Methanobacterium, Methanosarcina, Methanococcus, Methanomicrobium, Methanospirillum, Methanothermobacter, and Methanoculleus .

74. The method according to any one of claims 69 to 73, wherein the anaerobic decomposition vessel comprises at least one species of bacteria selected from the group consisting of: Pseudomonas aeruginosa, Pseudomonas putida, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Escherichia coli, Escherichia fergusonii, and Escherichia albertii, Zoogloea ramigera, Zoogloea caeni, and Zoogloea oryzae.

75. The method according to any one of claims 69 to 74, wherein the anaerobic decomposition vessel comprises a plurality of microorganisms, wherein the majority of the microorganisms are carbon decomposing bacteria, and wherein the plurality of microorganisms does not include ammonia decomposing bacteria.

76. The method according to any one of claims 69 to 75, wherein the first aerobic decomposition vessel comprises facultative anaerobic bacteria capable ofconsuming organic acids in low oxygen environments, selected from the group consisting of species from the genera Lactobacillus, Streptococcus, and Enterobacter.

77. The method according to any one of claims 69 to 76, wherein the first aerobic decomposition vessel comprises obligate anaerobic bacteria that can metabolize organic acids without the presence of oxygen, including species of obligate anaerobic bacteria from the genera Bacteroides, Clostridium, and Eubacterium.

78. The method according to any one of claims 69 to 77, wherein the first aerobic decomposition vessel does not comprise nitrification bacteria, such as Nitrosomonas or Nitrobacter, in amounts sufficient to maintain the desired low oxygen and non-nitrifying environment for the consumption of organic acids.

79. The method according to any one of claims 69 to 78, wherein the first aerobic decomposition vessel comprises a liquid medium maintained at a low dissolved oxygen (DO) level, specifically characterized by a dissolved oxygen concentration not exceeding 2 mg / L (2 ppm), and further comprises a bacterial composition adapted to thrive in such low oxygen conditions.

80. The method according to any one of claims 69 to 79, wherein the method is operated continuously, and further wherein the hydraulic retention time of the solution acidified with an acid in the oxidation reactor is sufficient to substantially decompose the acid, such as substantially decomposing the base component of acetic acid — acetate.

81. The method according to any one of claims 69 to 79, wherein the method is operated in batches, and wherein the hydraulic retention time of the solution acidified with the acid in the oxidation reactor is sufficient to substantially decompose the base component of the acid, such as substantially decomposing the base component of acetic acid — acetate.

82. The method according to any one of claims 69 to 81, wherein the biofertilizer is compatible as a component of YAN (Yeast Assimilable Nitrogen).

83. A liquid fertilizer produced by an organic waste digester, wherein the liquid fertilizer has a, and a biological oxygen demand (BOD) of less than 125 mg / L.

84. The liquid fertilizer of claim 83, wherein the liquid fertilizer is stable upon addition of peroxides.

85. The liquid fertilizer of claim 83 or 84, further comprising an organic acid.

86. The liquid fertilizer of any one of claims 83 to 85, further comprising a carbonaceous species to raise the C:N ratio, such as an organic acid selected from the group consisting of humic acid, fulvic acid, citric acid, and acetic acid.

87. The liquid fertilizer of any one of claims 83 to 86, wherein the composition is produced by introducing the wastewater into an anaerobic decomposition vessel configured to produce an anaerobic effluent; wherein the anaerobic decomposition vessel comprises an organic material, such as an organic material comprising a sugar, protein, lipid or cellulose, preferably a food or crop material and, wherein the anaerobic decomposition vessel is maintained at an anaerobic condition at a pH from 6.0 to 9.0; transferring the anaerobic effluent from the anaerobic decomposition vessel to a first aerobic decomposition vessel, preferably by a conduit, which connects the anaerobic decomposition vessel to the first aerobic decomposition vessel; digesting the anaerobic effluent in the first aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, while maintaining a pH from 7.0 to 9.0, and an oxidation reduction potential (ORP) between -1000 mV and 1000 mV to produce a first aerobic effluent; wherein the first aerobic vessel comprises at least one organic acid consuming organism, such as a bacteria from the genus Pseudomonas, Bacillus, Escherichia, or Zoogloea,' transferring the first aerobic effluent from the first aerobic decomposition vessel to a second aerobic decomposition vessel, preferably by a conduit, which connects the first aerobic decomposition vessel to the second aerobic decomposition vessel; and digesting the first aerobic effluent in the second aerobic decomposition vessel in the presence of oxygen or an oxidizer, such as hydrogen peroxide, or ozone, to produce a second aerobic effluent while maintaining a pH from 5.5 to 6.2 or 7.1 to 7.8; wherein the second aerobic decomposition vessel comprises at least one chemoautotrophic microorganism, such as a bacteria from the genus Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus, Nitrosococcus, or Nitrosospira, e.g., a bacterial population obtained from compost, liquid compost, or earthworm castings.

88. A method for conditioning a process fluid in a multi-stage biological treatment system comprising at least one aerobic decomposition vessel, the method comprising: fluidically connecting at least one base media contactor to the aerobic decomposition vessel; introducing the process fluid into the base media contactor comprising a packed bed of alkaline-reactive solids selected from calcite, dolomitic limestone, phosphate rock, magnesium oxide, or combinations thereof; allowing the process fluid to contact the packed bed in an upflow or downflow configuration; dissolving alkaline constituents from the solids into the fluid to raise pH and release mineral ions comprising at least calcium, magnesium, or phosphate; wherein the base media contactor is operable to modulate pH and supplement mineral ions that support microbial activity in the aerobic decomposition vessel and enhance the agronomic value of a resulting biofertilizer product.

89. The method according to any one of claims 1 to 81, and 88 wherein the base media contactor is configured in a downflow orientation and further operates as a physical filtration unit to remove suspended solids prior to subsequent filtration stages comprising microfiltration, ultrafiltration, or nanofiltration.

90. A method for maintaining microbial stability and continuous operation in a biological treatment system comprising at least one anaerobic decomposition vessel and at least two aerobic decomposition vessels, the method comprising: monitoring process parameters comprising FOS / TAC ratio, electrical conductivity, pH, bicarbonate concentration, ammonium concentration, and nitrite concentration; modulating solids loading and hydraulic retention time in the anaerobic decomposition vessel based on effluent transfer rates and monitored conditions; controlling aeration levels in a first aerobic decomposition vessel to limit ammonia volatilization and prevent unintended partial nitrification; adjusting dosing frequency and implementing pH cycling in a second aerobic decomposition vessel to manage nitrite accumulation and maintain nitrifying microbial viability;wherein the operational parameters of all treatment stages are dynamically coordinated through a closed-loop control framework.

91. The method according to any one of claims 1 to 81, and 88-90, further comprising adjusting exposure of process fluid to a base media contactor containing alkaline-reactive solids in response to elevated nitrite levels or declining bicarbonate concentrations in the second aerobic decomposition vessel.

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