Ammonia-oxidizing microorganisms for use and delivery to the digestive system

Abstract

Providing ammonia-oxidizing microorganisms for use and delivery to the digestive system. [Solution] Provided are ammonia-oxidizing microbial preparations for delivery to the digestive system, a kit containing the ammonia-oxidizing preparations for delivery to the digestive system, and a device for administering the ammonia-oxidizing preparations to the digestive system. A method for introducing ammonia-oxidizing microorganisms into the digestive system is provided. A method for treating disorders, including digestive disorders and inflammatory disorders, with the ammonia-oxidizing microbial preparations is provided. In some embodiments, the preparations may be food products. Food products may include foods, beverages, dietary supplements, functional foods, additives, or medical nutrition products.

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Patent Application No. 62 / 364,079, filed July 19, 2016, and U.S. Provisional Patent Application No. 62 / 397,710, filed September 21, 2016, under 35 U.S.C. § 119(e), the entire disclosures of each of these applications are incorporated herein by reference in their entirety for all purposes.

[0002] This embodiment relates, as a whole, to the restoration of microbiomes, and more specifically, to ammonia-oxidizing microorganisms associated with microbiomes. [Background technology]

[0003] Beneficial bacteria can be used to inhibit the growth of pathogenic bacteria. Bacteria and other microorganisms are ubiquitous in the environment. The discovery of pathogenic bacteria and the pathogenicity of disease have had a significant impact on health and disease states, and can be beneficial. Microorganisms are, for example, a normal part of the environment of all living things. In the gut, these bacteria are not pathogenic under normal conditions and actually improve health by making the normal contents of the intestines less suitable for organisms that cause disease. [Overview of the Initiative] [Means for solving the problem]

[0004] One or more embodiments disclose a method for introducing ammonia-oxidizing microorganisms (AOMs). This method may include enteral administration of a preparation containing AOMs.

[0005] In some embodiments, enteral administration may include oral administration, intraoral administration, sublabial administration, or sublingual administration.

[0006] One or more embodiments disclose a method for introducing ammonia-oxidizing microorganisms (AOMs). This method may include rectal administration of a preparation containing AOMs.

[0007] In some embodiments, rectal administration may include administration via suppositories or enemas. In other embodiments, rectal administration may involve a fecal microbiota transplantation procedure.

[0008] One or more embodiments disclose a method for colonizing a target digestive system. This method may include administering an effective amount of a preparation containing AOM that colonizes target tissue of the digestive system to the target digestive system.

[0009] One or more embodiments disclose a method for improving digestion in a subject. This method may include administering an effective amount of a preparation containing AOM to a subject to improve digestion in the subject.

[0010] One or more embodiments disclose a method for treating gastrointestinal disorders in a subject. This method may include administering an effective amount of a preparation containing AOM to the subject to treat the gastrointestinal disorder.

[0011] In some embodiments, a target percentage of the AOM is administered to the subject's digestive system. The amount and / or frequency of administration is sufficient to increase the viscosity of mucus in at least a portion of the subject's digestive system. Administration of the preparation may result in increased tolerance, decreased sensitivity, and / or improved nutrient uptake related to the food or beverage consumed by the subject.

[0012] In at least some embodiments, the preparation containing AOM is administered by ingestion into the subject's digestive system. The subject may have a substantially empty stomach when the preparation is administered. The preparation may be administered after administration of an antibiotic or a bowel cleansing preparation. The method may further include administering water to the subject after administration of the preparation.

[0013] In some embodiments, gastrointestinal disorders may be inflammatory conditions. For example, inflammatory conditions may include colitis, necrotizing enterocolitis, inflammatory bowel disease, ulcers, Crohn's disease, ulcerative colitis, celiac disease, gluten sensitivity, heartburn, pancreatitis, appendicitis, gastritis, gastroenteritis, irritable bowel syndrome, or dental or periodontal conditions. Inflammatory conditions may be associated with catheter-based delivery of substances, such as enteral nutrition. Inflammatory conditions may be infections with one or more of the following microorganisms: H. pylori, C. diff, cholera, amoebic dysentery, Y. enterocolitica, S. enteritidis, S. typhimurium, Shigella sonnei, and E. coli. In other embodiments, gastrointestinal disorders may be associated with lactose, food, or beverage intolerance, SIBO, malabsorption, biliary disorders, reflux, or dispoxia. Gastrointestinal disorders may be characterized by constipation or diarrhea. Gastrointestinal disorders may be characterized by hyperammonemia. Gastrointestinal disorders may include hepatic failure, such as acute or chronic hepatic failure.

[0014] In some embodiments, administration can reduce bloating, diarrhea, gas, stomach pain, stomach cramps, or borborygmus in the subject.

[0015] In some embodiments, administration may be before or after medical procedures, such as catheterization, endoscopy, or colonoscopy, or dental procedures. Administration may be via a device. The preparation may be administered before, during, or after the onset of a gastrointestinal condition. The preparation may be administered in response to gastrointestinal symptoms, triggers, or prodromal symptoms, such as discomfort or changes in bowel habits. The method may involve determining whether the subject requires treatment for a gastrointestinal disorder.

[0016] In some embodiments, administration may be performed directly to the deposit tissue or the target tissue. The deposit tissue, target tissue, or both may be the mucosa of the subject. The deposit tissue, target tissue, or both may be related to the stomach of the subject. The deposit tissue, target tissue, or both may be the salivary glands, oral cavity, pharynx, tongue, esophagus, liver, gallbladder, common bile duct, colon (transverse colon, ascending colon, and / or descending colon), cecum, appendix, rectum, anus, pancreas, pancreatic duct, large intestine, or small intestine (duodenum, jejunum, and / or ileum).

[0017] In some embodiments, the target tissue may be associated with a desired local effect. The target tissue may be associated with a desired systemic effect. For example, the desired systemic effect may involve the treatment of one or more of the following conditions: headache, cardiovascular disease, inflammation, immune response, autoimmune disorder, liver disease, infection, neurological disorder, psychiatric disorder, nitric oxide disorder, urea cycle disorder, congestion, vasodilation disorder, skin disease, wound healing, reaction to insect bites, eye disorder, connective tissue disorder, and infection with certain viruses, bacteria, or fungi.

[0018] In some embodiments, administration of an effective dose of the preparation may promote endothelial function. Administration of an effective dose of the preparation may alter or change the levels of nitrite or NO in the target tissue or circulation. Administration of an effective dose of the preparation may modulate the microbiome associated with the target digestive system.

[0019] In some embodiments, the preparation may be administered as a solution, suspension, emulsion, ointment, gel, hydrogel, or liquid, for example, as a drop, spray, aerosol, or mist. The preparation may be formulated as a tablet or capsule. The preparation may contain microspheres or microcapsules. The preparation may be formulated to be adapted to the target gastrointestinal system. The preparation may be formulated for immediate release or extended release. The preparation may be formulated to deliver nitrite or NO to target tissue, topically or systemically. The preparation may be formulated for transmucosal delivery and / or to enter circulation, for example, topically or systemically.

[0020] In some embodiments, the treatment method may further comprise administering a second amount of the preparation to the subject. In at least some embodiments, the preparation can be administered as part of a combination therapy. The method may further comprise administering a second treatment in combination with the preparation. The preparation can be administered over a period of time before the initiation of the second treatment, simultaneously with the second treatment, or over a period of time after the discontinuation of the second treatment. The second treatment can be administered via an alternative mode of administration, such as inhalation or intranasal techniques. The subject may receive a therapeutic level of the second treatment. The preparation can be administered together with an anti-inflammatory agent. The preparation can be administered together with a medical approach that treats a related disease or disorder, or the symptoms of a related disease or disorder, for example, that is approved to treat or is commonly used to treat it. The preparation can be administered before or after a surgical or diagnostic procedure. The second treatment may comprise a surgical procedure. The preparation can be administered together with exercise, fiber, laxatives, antidiuretics, probiotics, therapeutic agents, or stress management. In at least some embodiments, the preparation can be administered in combination with a therapeutic treatment for colitis, necrotizing enterocolitis, inflammatory bowel disease, ulcers, Crohn's disease, ulcerative colitis, celiac disease, gluten hypersensitivity, heartburn, pancreatitis, appendicitis, gastritis, gastroenteritis, or irritable bowel syndrome. The preparation can be administered together with nitrites, nitrates, and / or NO.

[0021] In some embodiments, the effective amount is a therapeutically effective dosage of AOM. The therapeutically effective dosage of AOM is about 1×10 3 、10 4 、10 5 、10 6 、10 7 、10 8 、10 9 、10 10 、10 11 、10 12 、10 13 、or 10 14The preparation is CFU or greater. The preparation may be administered as an analgesic and / or prophylactic agent. The preparation may be self-administered. The preparation may be administered approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 times per day. The preparation may be administered over approximately 1–3, 3–5, 5–7, 7–9, 5–10, 10–14, 12–18, 12–21, 21–28, 28–35, 35–42, 42–49, 49–56, 46–63, 63–70, 70–77, 77–84, or 84–91 days. The preparation can be administered within 30, 60, 90, 120, 150, or 180 minutes after the subject wakes up. The preparation can be administered within 30, 60, 90, 120, 150, or 180 minutes before the subject goes to sleep. The preparation can be administered within 30, 60, 90, 120, 150, or 180 minutes before the subject eats a meal. The preparation can be administered within 30, 60, 90, 120, 150, or 180 minutes before the subject washes or showers.

[0022] In some aspects, the subjects may be female. In other aspects, the subjects may be male. The subjects may be characterized as one of the following ethnic / racial groups: Asian, Black or African American, Hispanic or Latino, White, or multi-ethnic. The subjects may have a disturbed microbiome. The subjects may be under 1 year old, or between 1 and 5 years old, between 5 and 10 years old, between 10 and 20 years old, between 20 and 30 years old, between 30 and 40 years old, between 40 and 50 years old, between 50 and 60 years old, or over 60 years old.

[0023] In some embodiments, the preparation may contain AOM in a buffer solution, for example, an aqueous buffer solution. The buffer solution, for example, an aqueous buffer solution, contains disodium phosphate and magnesium chloride, for example, 50 mM Na2HPO4 and 2 mM MgCl2 in water. The buffer solution, for example, an aqueous buffer solution, essentially consists of disodium phosphate and magnesium chloride, for example, 50 mM Na2HPO4 and 2 mM MgCl2 in water. The buffer solution, for example, an aqueous buffer solution, consists of disodium phosphate and magnesium chloride, for example, 50 mM Na2HPO4 and 2 mM MgCl2 in water. The preparation may be characterized by a physiological pH level. The preparation may further contain, or be administered simultaneously with, compounds that promote the proliferation or metabolism of AOM, NO production, and / or urease activity. In at least some embodiments, the preparation may contain at least one of ammonia, ammonium salts, and urea. The preparation may contain a controlled-release material, for example, a delayed-release material. The preparation may further contain additives, such as pharmaceutically acceptable additives. These additives may include absorption enhancers or osmotic enhancers, preservatives, antioxidants, buffers, chelating agents, ion exchangers, solubilizers, suspending agents, thickeners, surfactants, wetting agents, isotonic agents, enzyme inhibitors, or vehicles for appropriate drug delivery. The additives may also include anti-adhesives, binders, coatings, disintegrants, fillers, fragrances, colorants, lubricants, flow enhancers, adsorbents, preservatives, or sweeteners. In at least some embodiments, the preparation may include mucosal adhesives, disintegrants, chelating agents, coatings, release regulators, or fillers. The preparation does not need to substantially contain other organisms. The preparation may further contain other organisms, such as biological communities.

[0024] In some embodiments, the preparation is approximately 1 × 10 3 From CFU / mL to approximately 1 × 10 14 The preparation may contain AOM between CFU / mL. 9 From CFU / mL to approximately 10 × 10 9AOM may contain ammonia-oxidizing bacteria (AOB) between CFU / mL. AOM may contain ammonia-oxidizing bacteria (AOB). In some embodiments, AOM may essentially consist of AOB. In at least some embodiments, AOM may consist of AOB. AOM may contain Nitrosomonas, Nitrosococcus, Nitrosospira, Nitrosocystis, Nitrosolobus, Nitrosovibrio, and combinations thereof. AOM may be Nitrosomonas eutropha (N. eutropha). AOM may be N. eutropha D23 with ATCC accession number PTA-121157. In at least some embodiments, AOM may contain ammonia-oxidizing archaea (AOA). AOM can convert ammonia or ammonium to nitrite at a rate of at least about 1 pmol / min / mg protein. AOM can convert ammonia or ammonium to nitrite at a rate of at least about 0.1 nmol / min / mg protein.

[0025] In some embodiments, the preparation can be administered, for example, by ingestion into a first tissue, such as a depositional tissue. The first tissue may be a target tissue. In at least some embodiments, the first tissue is other than the target tissue, for example, the preparation may be applied to the first tissue, and the preparation or a product of the preparation, such as NO, may be transported to a second tissue, such as a target tissue, for example, by diffusion.

[0026] In some embodiments, biome-inert products may be used in conjunction with administered preparations containing AOM.

[0027] According to one or more embodiments, the preparation containing AOM described in any one of the preceding claims may be for enteral administration to a subject.

[0028] In some embodiments, the preparation may be a food product. The food product may include food, beverage, dietary supplement, functional food, additive, or medical nutrition product.

[0029] According to one or more embodiments, the preparation may contain AOM. The preparation may be intended to treat gastrointestinal disorders in a subject.

[0030] In some embodiments, the preparation may be packaged for single use. In some embodiments, the preparation may be packaged for multiple uses.

[0031] According to one or more embodiments, the device may be configured to deliver a preparation containing AOM to target tissue of the target digestive system.

[0032] According to one or more embodiments, the kit may include a preparation containing AOM for, for example, delivery to the digestive system of a subject or for treating a digestive disorder in a subject.

[0033] This disclosure intends to include any one or more combinations of the embodiments described above, as well as any one or more of the embodiments described in the Detailed Description and any Examples. [Modes for carrying out the invention]

[0034] According to one or more embodiments, the Disclosure provides various methods or forms for introducing ammonia-oxidizing microorganisms into a target. These methods or forms involve administering ammonia-oxidizing microorganisms, for example, a preparation, composition, formulation, or product containing ammonia-oxidizing microorganisms, to a target. In at least some embodiments, the ammonia-oxidizing microorganisms can therefore typically be restored to the target microbiome. In at least some embodiments, the ammonia-oxidizing microorganisms may include or be essentially live ammonia-oxidizing microorganisms.

[0035] Preparations, compositions, and / or formulations containing, essentially, or consisting of ammonia-oxidizing microorganisms are disclosed, including, for example, cosmetic products, therapeutic products, consumer products, non-natural products, natural products, and enhanced natural products. These preparations, compositions, and / or formulations are disclosed herein for use in various applications, e.g., cosmetic and / or therapeutic applications. The preparations, compositions, and / or formulations can be administered in an effective amount for their intended use, e.g., for cosmetic or therapeutic applications. Preparations, compositions, and / or formulations containing ammonia-oxidizing microorganisms are provided for various modes of administration to a subject. Preparations, compositions, and / or formulations containing ammonia-oxidizing microorganisms are provided for use in treating various conditions and / or disorders in a subject. Methods for treating a subject for various conditions and / or disorders via the administration of ammonia-oxidizing microorganisms are disclosed. Devices for use in the administration of ammonia-oxidizing microorganisms to a subject are also provided.

[0036] microbiology According to one or more embodiments, essentially any ammonia-oxidizing microorganism (AOM) can be used or implemented. The ammonia-oxidizing microorganism may typically be inorganic nutrient. The ammonia-oxidizing microorganism can produce nitrite and / or nitric oxide from ammonia.

[0037] The properties of inorganic vegetative ammonia-oxidizing bacteria (AOBs) are well described, for example, by Whitlock in U.S. Patent No. 7,820,420. Since that filing, the class of inorganic vegetative microorganisms that oxidize ammonia for ATP production has been expanded to include ammonia-oxidizing archaea (AOAs), with archaea being excluded from the class of bacteria and placed in its own class. For the purposes of this disclosure, any and all inorganic vegetative ammonia-oxidizing microorganisms that share the property of oxidizing ammonia to produce ATP can be executed. AOMs, including both AOBs and AOAs, share the essential property of oxidizing ammonia to NO and nitrites, and all known AOMs lack the ability to use organic substrates for ATP production. Bacteria can utilize ammonia at high concentrations, while archaea can utilize ammonia at low concentrations. The physiological levels of ammonia are within the range that both bacteria (AOBs) and archaea (AOAs) can utilize. Throughout this disclosure, all references to ammonia-oxidizing bacteria, in particular, should be considered equally applicable to all ammonia-oxidizing microorganisms, such as all ammonia-oxidizing archaea, and all these terms may be used interchangeably herein.

[0038] Ammonia-oxidizing bacteria (AOBs) are ubiquitous Gram-negative obligate bacteria with the unique ability to produce energy solely from the conversion of ammonia to nitrite. In some embodiments, ammonia-oxidizing bacteria (AOBs) of the genus Nitrosomonas are Gram-negative obligate inorganic (chemosynthetic autotrophic) bacteria with the unique ability to produce nitrite and nitric oxide from ammonia alone as an energy source. They are widely present in both soil and aquatic environments and are essential components of environmental nitrification processes. These bacteria possess advantageous properties related to various cosmetic and therapeutic uses, for example, according to one or more embodiments described herein. While we do not wish to be bound by any particular logic, due to the roles of nitrite and nitric oxide on human skin as important components of several physiological functions such as vasodilation, skin inflammation, and wound healing, these bacteria may possess various beneficial properties for both healthy and immunopathological states. These bacteria are safe for human use because they grow slowly, cannot grow on organic carbon sources, may be susceptible to soap and antibiotics, and are not associated with any disease or infection in animals or humans.

[0039] Ammonia-oxidizing microorganisms produce coenzyme Q8 (CoQ8) as a byproduct of the process by which they produce nitrite and nitric oxide. CoQ8 is a coenzyme Q with eight carbon atoms in its isoprenoid side chain. While we do not wish to be bound by any particular theory, the favorable properties of these microorganisms may be further enhanced by their specific ability to produce CoQ8, due to the role of coenzyme Q as a key component of several cellular functions, such as mediating cell signaling and preventing cell death (anti-aging).

[0040] In some embodiments, ammonia-oxidizing bacteria can catalyze the following reactions.

[0041] At neutral pH levels, ammonia, which is produced from ammonium near neutral pH conditions, is the substrate for the initial reaction. The conversion of ammonia to nitrite occurs in two steps, catalyzed by ammonia monooxygenase (AMO) and hydroxylamine oxidoreductase (HAO), respectively, as follows: NH3+2H + +2e- +O2→NH2OH+H2O (A) NH2OH + H2O → NO2 - +4e-+5H + (B)

[0042] In some cases, reaction B is reported to produce nitrite (HNO2) at low pH, as follows: NH2OH + H2O → HNO2 + 4e- + 4H +

[0043] In a particular embodiment, NH4 + And NH3 may be used interchangeably throughout this disclosure.

[0044] Examples of ammonia-oxidizing bacteria include strains of Nitrosomonas eutropha, such as D23 and C91, which are discussed herein, as well as other bacteria of the genera Nitrosomonas, Nitrosococcus, Nitrosospira, Nitrosocystis, Nitrosolobus, and Nitrosovibrio. Strain D23 of Nitrosomonas eutropha refers to the strain designated AOB D23-100, deposited on April 8, 2014, with accession number PTA-121157 at the American Tissue Culture Collection (ATCC) (10801 University Blvd., Manassas, VA, USA). The nucleic acid sequence of accession number PTA-121157, such as the genome sequence, is incorporated herein by reference in its entirety for all purposes. "AOB D23-100" may also be referred to as D23 or B244 throughout this disclosure.

[0045] Examples of ammonia-oxidizing archaeons include archaeons of the genera Methanobrevibacter, Methanosphaera, Methanosarcina, Nitroscaldus, Nitrosopumilus, and Nitrososphaera (e.g., Nitrososphaera viennensis, Nitrososphaera gargensis). Archaeons of different phylotypes, such as methanogenic and halphilic archaeons, may be included in the preparations disclosed herein. Examples of archaeons further include archaeons of the Euryarchaeota phyla (e.g., Methanosarcina), Crenarchaeota phyla, Aigarchaeota phyla, and Thaumarchaeota phyla (e.g., Giganthauma karukerense, Giganthauma insulaporcus, Caldiarchaeum subterraneum, Cenarchaeum symbiosum).

[0046] Each and all nucleic acid sequences and amino acid sequences disclosed in International (PCT) Patent Application Publication WO2015 / 160911 (International (PCT) Patent Application PCT / US2015 / 025909, filed on 15 April 2015) are incorporated herein by reference in their entirety for all purposes. Similarly, any ammonia-oxidizing bacteria disclosed in International (PCT) Patent Application Publication WO2015 / 160911 (International (PCT) Patent Application PCT / US2015 / 025909, filed on 15 April 2015) are also incorporated herein by reference in their entirety for all purposes. In certain embodiments, the ammonia-oxidizing microorganism is the strain described in these documents.

[0047] According to one or more embodiments, ammonia-oxidizing microorganisms may exist in several metabolic states, such as growth, storage, and / or polyphosphate-loaded states.

[0048] According to one or more embodiments, the ammonia-oxidizing microorganism may have optimized properties, such as the ability to inhibit the growth of pathogenic bacteria, and an enhanced ability to produce nitric oxide and nitric oxide precursors.

[0049] Optimized Nitrosomonas eutropha (N. eutropha) is, when used herein, an optimized growth rate, optimized NH4 + Oxidation rate, and / or optimization of NH4 + This refers to N. eutropha with resistance. In one embodiment, this refers to at least one nucleotide, for example, ammonia monooxygenase, hydroxylamine oxidoreductase, cytochrome c554, and cytochrome c M The nucleotides in the selected gene differ from those in naturally occurring N. eutropha. This difference may arise, for example, from spontaneous mutations, induced mutations, or targeted genetic engineering selections in N. eutropha. In one embodiment, this differs from naturally occurring N. eutropha in that it has a set of alleles that do not exist together in nature. These differences may provide one or more of the treatment or prevention of diseases or conditions, such as diseases or conditions associated with low nitrite levels, but are not limited to these.

[0050] Any ammonia-oxidizing bacteria, such as N. eutropha, for example, N. eutropha known as "D23," also known as "B244" or "AOB D23-100," may possess some of the characteristics described above. Any ammonia-oxidizing archaea (AOA) may also possess some of the characteristics described above.

[0051] The AOBs considered in this disclosure may include mutations relative to wild-type AOBs. These mutations may occur, for example, spontaneously, be introduced by random mutagenesis, or be introduced by targeted mutagenesis. For example, an AOB may lack one or more genes or regulatory DNA sequences typically present in a wild-type AOB. An AOB may also include point mutations, substitutions, insertions, deletions, and / or rearrangements relative to a sequenced strain or a wild-type strain. An AOB may be a purified preparation of an optimized AOB.

[0052] In certain embodiments, AOB is transgenic. For example, it may include one or more genes or regulatory DNA sequences that are lacking in wild-type ammonia-oxidizing bacteria. More specifically, the ammonia-oxidizing bacteria may include, for example, a reporter gene, a selection marker, a gene encoding an enzyme, or a promoter (including inductive or repressive promoters). In some embodiments, the added gene or regulatory DNA sequence is incorporated into the bacterial chromosome; in some embodiments, the added gene or regulatory DNA sequence is located on a plasmid.

[0053] In some embodiments, AOB differs from naturally occurring bacteria in at least one nucleotide. For example, AOB may differ from naturally occurring bacteria in genes or proteins that are part of the relevant pathway, such as the ammonia metabolic pathway, the urea metabolic pathway, or the pathway for producing nitric oxide or a nitric oxide precursor. More specifically, AOB may include mutations that increase the activity of the pathway, for example, by increasing the level or activity of elements of the pathway.

[0054] The aforementioned mutations can be introduced using any suitable technique. Numerous methods for introducing mutations at a given location are known. For example, site-directed mutagenesis, oligonucleotide-directed mutagenesis, or site-specific mutagenesis can be used. Non-exclusive examples of specific mutagenesis protocols are described, for example, in Mutagenesis, pp. 13.1–13.105 (Sambrook and Russell, eds., Molecular Cloning: A Laboratory Manual, Vol. 3, 3rd edition supplement, 2001). In addition, non-limiting examples of well-characterized mutagenesis protocols available from commercial suppliers include: Altered Sites® II in vitro Mutagenesis Systems (Promega Corp., Madison, Wis.); Erase-a-Base® System (Promega, Madison, Wis.); GeneTailor® Site-Directed Mutagenesis System (Invitrogen, Inc., Carlsbad, Calif.); and QuikChange® II Site-Directed Mutagenesis. This includes, but is not limited to, Kits (Stratagene, La Jolla, Calif.) and Transformer® Site-Directed Mutagenesis Kits (BD-Clontech, Mountain View, Calif.).

[0055] In certain embodiments of this disclosure, the ammonia-oxidizing bacteria may be in pure culture. Preparations (formulations or compositions) of ammonia-oxidizing microorganisms may contain, be essentially derived from, or consist of pure cultures of ammonia-oxidizing microorganisms.

[0056] The ammonia-oxidizing bacteria of this disclosure may be derived from a genus selected from the group consisting of Nitrosomonas, Nitrosococcus, Nitrosospira, Nitrosocystis, Nitrosolobus, Nitrosovibrio, and combinations thereof.

[0057] This disclosure provides, in particular, a specific, for example, optimized strain of ammonia-oxidizing bacteria, N. eutropha strain D23, which can increase the production of nitric oxide and nitric oxide precursors on the surface of a subject, for example, a human subject. This disclosure also provides the bacteria, as well as preparations, compositions, formulations, and products containing the bacteria, and methods for administering and using them.

[0058] In several embodiments, the ammonia-oxidizing bacteria, such as N. eutropha, do not exist in nature. For example, it may accumulate the desired mutation during the selection period. In other embodiments, the desired mutation may be introduced by the experimenter. In some embodiments, N. eutropha may be a purified preparation or an optimized N. eutropha.

[0059] In preferred embodiments, the N. eutropha strain is autotrophic and therefore cannot cause infection. The preferred strain utilizes urea and ammonia, and as a result, hydrolysis of urea in sweat is not required before absorption and utilization by the bacteria. Also, to grow at low pH, the bacteria utilize NH4 + It can absorb ions or urea. The selected strain should also be viable on a subject, such as human skin, and should be able to withstand the conditions there.

[0060] While this disclosure refers in detail to N. eutropha strain D23, preparations, methods, compositions, treatments, formulations, and products using one or more other strains of N. eutropha, one or more other species of Nitrosomonas, and one or more other ammonia-oxidizing microorganisms, such as ammonia-oxidizing bacteria or other ammonia-oxidizing archaea, may be used.

[0061] In certain embodiments, bacteria having the above-described sequence characteristics have (1) an optimized growth rate measured by doubling time, (2) an optimized growth rate measured by OD600, and (3) an optimized NH4 + Oxidation rate, (4) Optimized NH4 + Resistance, and (4) Optimization NO2 - It possesses one or more of the following resistances. Specific non-exclusive partial combinations of these properties are identified in the following paragraph.

[0062] In some embodiments, the ammonia-oxidizing bacteria described herein, such as N. eutropha, are: (1) optimized growth rate measured by doubling time, (2) optimized growth rate measured by OD600, and (3) optimized NH4 + Oxidation rate, (4) Optimized NH4 + Resistance, and (4) Optimization NO2 -A bacterium may have one or more resistances. For example, a bacterium may have properties (1) and (2); (2) and (3); (3) and (4); or (4) and (5) from the list at the beginning of this paragraph. Another example is a bacterium which may have properties (1), (2), and (3); (1), (2), and (4); (1), (2), and (5); (1), (3), and (4); (1), (3), and (5); (1), (4), and (5); (2), (3), and (4); (2), (3), and (5), or (3), (4), and (5). As a further example, a bacterium may have characteristics (1), (2), (3), and (4) from the list at the beginning of this paragraph; (1), (2), (3), and (5); (1), (2), (4), and (5); (1), (3), (4), and (5); or (2), (3), (4), and (5). In some embodiments, a bacterium has characteristics (1), (2), (3), (4), and (5) from the list at the beginning of this paragraph.

[0063] In certain embodiments, the N. eutropha strain includes a nucleic acid sequence, e.g., a genome, that hybridizes under low, medium, high, or very high stringency conditions, or other hybridization conditions, to the genome of the D23 strain designated AOB D23-100, deposited with the ATCC Patent Depository on April 8, 2014, in the form of 25 vials, under Sequence ID 1 of International (PCT) Patent Application Publication WO2015160911 (International (PCT) Patent Application No. PCT / US2015 / 025909, filed on April 15, 2015), or accession number PTA-121157, or to their complements.

[0064] The D23 strain is not thought to be a naturally occurring product, but rather acquired specific mutations and characteristics during long-term culture and selection in the laboratory. For example, D23 has NH4 levels greater than approximately 200 or 250 mM. + Under these conditions, it has the ability to multiply for more than 24 hours.

[0065] In some embodiments, the N. eutropha disclosed herein differ from naturally occurring bacteria in terms of siderophore abundance. For example, N. eutropha may have elevated or reduced siderophore levels compared to N. eutropha C91. Generally, siderophores are secreted iron chelate compounds that help bacteria capture iron from their environment. Siderophores may be peptides or small organic molecules.

[0066] The implementation of this invention may, unless otherwise specified, employ conventional methods of immunology, molecular biology, and recombinant DNA techniques within the scope of the skills of those skilled in the art. Such techniques are well described in the literature. For example, Sambrook et al., Molecular Cloning: Please refer to A Laboratory Manual (latest edition) and Current Protocols in Molecular Biology (FMAusubel et al., latest edition).

[0067] Selection Definition Ammonia-oxidizing microorganisms, such as ammonia-oxidizing bacteria, refer to microorganisms that can oxidize ammonia or ammonium to nitrite at a certain rate, such as a substantial rate, such as a predetermined rate. The rate, such as a predetermined rate, means, for example, in The ammonium ion (NH4) is determined or measured by a vitro assay, or when administered to a subject, such as a human. + (For example, from about 200 mM) nitrite (NO2 - This may refer to the conversion to NO2. For example, for a continuous culture with an OD of about 0.5, the rate is at least about 1 picomole per minute per 1 mg of protein, and 0.01, 0.1, 1, 10, 25, 50, 75, 125, or 150 nanomoles per minute per 1 mg of protein. -For example, approximately 0.01-1, 0.1-50, 50-100, 100-150, 75-175, 75-125, 100-125, 125-150, or 125-175 nanomoles / min / mg of protein, for example, approximately 125 nanomoles of NO2 per minute per 1 mg of protein. - The conversion rate may be between approximately 1 picomole per minute per mg of protein and approximately 1 millimoles per minute per mg of protein. The conversion rate is up to approximately 1 mole of NO2 per minute per mg of protein. - For example, per 1 mg of protein, at least approximately, approximately, or up to approximately 1 decimol, 1 centimole, 1 millimol, or 1 micromol of NO2 per minute. - It is possible.

[0068] As used herein, “pure culture” means a composition containing organisms that are substantially free of other organisms. For example, a pure culture of ammonia-oxidizing bacteria is a culture that is substantially free of organisms other than ammonia-oxidizing bacteria. For example, a pure culture of N. eutropha is a culture that is substantially free of organisms other than N. eutropha. In some embodiments, “substantially free” indicates that it is undetectable by methods used to detect other organisms, such as plating the culture and examining the colony morphology, or by PCR for conserved genes such as 16S RNA. A pure culture composition may contain non-living elements, such as nutrients or additives. Any embodiment, preparation, composition, or formulation of ammonia-oxidizing bacteria discussed herein may optionally contain, be essentially, or be derived from a pure culture of ammonia-oxidizing bacteria.

[0069] Throughout this disclosure, "formulation" may refer to a composition, preparation, or product.

[0070] As used herein, “autotrophic organism,” for example, “autotrophic bacteria,” is any organism capable of self-nutrition by using inorganic materials as a nutrient source and photosynthesis or chemosynthesis as an energy source. Autotrophic bacteria utilize carbon dioxide and other sources, including the oxidation of ammonia to nitrite, the oxidation of hydrogen sulfide, and Fe 2+ Fe 3+ Organic compounds can be synthesized from ATP derived from oxidation. The autotrophic bacteria of this disclosure are incapable of causing infection.

[0071] As used herein, “combined” administration means that two (or more) different treatments are delivered to a subject during the course of the subject’s illness, for example, that two or more treatments are delivered after the subject has been diagnosed with the illness and before the illness is cured or eliminated. In some embodiments, the delivery of one treatment is still taking place when the delivery of a second treatment begins, resulting in overlap. This may be referred to herein as “simultaneous,” “combined,” or “parallel delivery.” In other embodiments, the delivery of one treatment is completed before the delivery of the other treatment begins. This may be referred to herein as “sequential” or “sequential delivery.” In some embodiments of any of these cases, the treatments are more effective for combined administration. For example, the second treatment is more effective to a greater extent than would be seen if the second treatment were administered in the absence of the first treatment, for example, a similar effect would be seen with fewer second treatments, or the second treatment would reduce symptoms, or a similar situation would be seen with the first treatment. In some embodiments, the delivery is such that the reduction of symptoms or other parameters related to the disorder exceeds that observed with a single treatment delivered in the absence of other treatments. The effects of the two treatments may be partially additive, fully additive, or more than additive (i.e., synergistic). The delivery may be such that the effect of the first treatment delivered is still detectable when the second treatment is delivered. In some embodiments, one or more treatments may be delivered before the patient is diagnosed with the disorder.

[0072] As used herein, the term “isolated” refers to material that has been removed from its original or native environment (e.g., its natural environment if it exists in nature). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide that has been separated from some or all of the coexisting material in a natural system by human intervention is isolated. Such polynucleotide may be part of a vector, and / or such polynucleotide or polypeptide may be part of a composition, and such vector or composition is still isolated in that it is not part of the environment in which it is found in nature.

[0073] As used herein, the term “optimized growth rate” means one or more of the following: doubling times of less than approximately 4, 5, 6, 7, 8, 9, or 10 hours when cultured under the batch conditions described in Example 2 herein; doubling times of less than approximately 16, 18, 20, 22, 24, or 26 hours when grown under the chemostat conditions described in Example 2 herein; or growth from an OD600 of about 0.15 to at least about 0.3, 0.4, 0.5, 0.6, 0.7, or 0.8 over about 1 or 2 days. In one embodiment, the optimized growth rate is a rate having a doubling time at least 10, 20, 30, 40, or 50% shorter than the doubling time of naturally occurring N. eutropha.

[0074] When used herein, "Optimized NH4" + "Oxidation rate" refers to the oxidation rate of at least approximately 50, 75, 125, or 150 micromoles of NH3 or NH4 per minute. + From NO2 - This refers to the rate of conversion to NH4. For example, the rate is at least approximately 50, 75, 125, or 150 micromoles per minute. + (For example, from NO2 at approximately 200 mM) -It may also be a conversion to NH4. In one embodiment, optimized NH4 + The oxidation rate is NH3 or NH4 + However, NO2 fertilization occurs at least 10, 20, 30, 40, or 50% more rapidly than in naturally occurring N. eutropha. - This is the speed at which it is converted.

[0075] When used herein, "Optimized NH4" + "Tolerance" refers to NH3 or NH4 levels above 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, or 300 mM. + This refers to the ability to grow for at least about 24 hours or 48 hours under these conditions. In one embodiment, optimized NH4 + Tolerance refers to the selected concentration of NH3 or NH4 + This refers to the ability to grow at least 10, 20, 30, 40, or 50% faster, or at least 10, 20, 30, 40, or 50% longer, than naturally occurring N. eutropha can grow in the presence of [unspecified substance].

[0076] As used herein, “transgenic” means containing one or more exogenous DNA portions. The exogenous DNA is derived from another organism, such as another bacterium, bacteriophage, animal, or plant.

[0077] As used herein, treatment of a disease or condition means reducing the severity or frequency of at least one symptom of that disease or condition compared to a similar but untreated patient. Treatment may also mean stopping, slowing, or reversing the progression of the disease or condition compared to a similar but untreated patient. Treatment may also include addressing the underlying causes of the disease and / or one or more symptoms.

[0078] As used herein, a therapeutically effective dose means a dose sufficient to prevent the progression of a disease or condition, to induce its regression, to alleviate the symptoms of a disease or condition, or to achieve a desired outcome. A therapeutically effective dose may be expressed, for example, by the number of bacteria or the number of viable bacteria (e.g., in units of CFU), or by the mass of bacteria (e.g., in units of milligrams, grams, or kilograms), or by the volume of bacteria (e.g., mm³). 3 It can be measured in units of measurement.

[0079] As used herein, the term “viability” refers to the ability of autotrophic bacteria, such as ammonia-oxidizing bacteria, to oxidize ammonia, ammonium, or urea to nitrite at a predetermined rate. In some embodiments, the rate is defined as at least about 1 picomole, 0.01, 0.1, 1, 10, 25, 50, 75, 125, or 150 nanomoles of NO2 per minute. - For example, approximately 0.01-1, 0.1-50, 50-100, 100-150, 75-175, 75-125, 100-125, 125-150, or 125-175 nanomoles / min, for example, about 125 nanomoles of NO2 per minute. - At the rate of, ammonium ions (NH4 + (For example, from about 200 mM) nitrite (NO2 - This can refer to the conversion to NO2. The conversion rate is up to approximately 1 mole per minute. - For example, at least about, approximately, or up to about 1 decimol, 1 centimole, 1 millimol, or 1 micromol of NO2 per minute. - This is possible. Live ammonia-oxidizing microorganisms may typically contain cultureable AOM, or AOM that can otherwise produce NO, nitrates, or nitrites.

[0080] As used herein, “Subject” may include animals, mammals, humans, non-human animals, domesticated animals, or companion animals. The term “Subject” is intended to include humans and non-human animals, e.g., vertebrates, megazoans, and primates. In certain embodiments, the subject is a mammalian subject, and in certain embodiments, the subject is a human subject. While applications in humans are explicitly anticipated, veterinary applications in non-human animals, for example, are also assumed herein. The term “non-human animals” in this disclosure includes all vertebrates, e.g., non-mammals (birds, e.g., chickens; amphibians; reptiles, etc.) as well as mammals, e.g., non-human primates, domesticated, and agriculturally useful animals, e.g., sheep, dogs, cats, cattle, pigs, and rats.

[0081] The term "microbiome" refers to a population of microorganisms, such as one or more species, that live on the surface of an object, for example, in the intestines, mouth, skin, and / or other locations within the object. The population may have one or more beneficial functions and / or benefits in relation to supporting life in the object.

[0082] "Biome-safe" means any object, such as a product, such as a cosmetic product, such as a finished cosmetic product, that can allow for minimal disturbance to the microbiome in question. For example, "biome-safe" means a product that can be applied to a target that allows the microbiome at the time of application to be maintained, minimally disturbed, and / or to return to that microbiome after a period of time following the application of the product. In embodiments, "biome-safe" may mean being harmless to ammonia-oxidizing microorganisms, for example, being harmless to ammonia-oxidizing bacteria in the sense that the product can allow for minimal disturbance to the ammonia-oxidizing bacteria in question. In embodiments, "biome-safe" may mean "biome-compatible".

[0083] "Natural products" are or may include products that are at least partially of natural origin. This may include any substance produced by a living organism, or any substance produced by a living organism, or the organism itself. Natural products may include whole organisms and parts of organisms (e.g., plant leaves), extracts of organisms, organic compounds of organisms, and purified organic compounds of organisms. Natural products may include or include found organic substances and cells, including primary metabolites (amino acids, carbohydrates, and nucleic acids) and secondary metabolites (organic compounds found in a limited range of species, e.g., polyketides, fatty acids, terpenoids, steroids, phenylpropanoids, alkaloids, specific amino acids and peptides, and specific carbohydrates). Natural products may include or be polymeric organic materials, e.g., cellulose, lignin, and proteins.

[0084] As used herein, “presence” or “level” may refer to one or more qualitative or quantitative amounts of any component, such as ammonia-oxidizing microorganisms, ammonia, ammonium ions, urea, nitrites, or nitric oxide. Presence or level may include zero or the absence of any component.

[0085] As used herein, the term “surfactant” includes compounds that can reduce surface tension, or interfacial tension, between two liquids or between a liquid and a solid. Surfactants can act as detergents, wetting agents, emulsifiers, foaming agents, and dispersants. Surfactants may include one or more of the following, alone or in combination with other surfactants or surfactant-like compounds: cocamidopropyl betaine (ColaTeric COAB), polyethylene sorbitol ester (e.g., Tween 80), ethoxylated lauryl alcohol (RhodaSurf 6 NAT), sodium laureth sulfate / lauryl glucoside / cocamidopropyl betaine (Plantapon 611 L UP), sodium laureth sulfate (e.g., RhodaPex ESB 70 NAT), alkyl polyglucoside (e.g., Plantaren 2000) Dr. Bronner's Castile Soap, Dr. Bronner's Baby Soap, Lauramine Oxide (ColaLux Lo), Sodium Dodecyl Sulfate (SDS), Polysulfonate Alkyl Polyglucoside (PolySufanate 160 P), Sodium Lauryl Sulfate (Stepanol-WA Extra K), and combinations thereof. Dr. Bronner's Castile Soap and Baby Soap contain water, organic coconut oil, potassium hydroxide, organic olive oil, organic hemp oil, organic jojoba oil, citric acid, and tocopherol. The surfactant may include sodium lauryl glucoside hydroxypropyl sulfonate (Suga® nate 160NC), lauramidopropyl betaine (Cola® Teric LMB); cocamidopropyl hydroxysultaine (Cola® Teric CBS); disodium cocoamphodiacetate (Cola® Teric CDCX-LV); and sodium lauryl glucoside hydroxypropyl phosphate (Suga® Fax D12). The surfactant may also include sodium lauroyl methyl isethionate (Iselux® LQ-CLR-SB); sodium cocoyl methyl taurate (Pureact WS Conc.); water (and) sodium lauroyl methyl isethionate (and) cocamidopropyl betaine (and) sodium cocoyl isethionate (and) sodium oleoyl methyl taurate (Iselux® SFS-SB). Other surfactants are intended by this disclosure.

[0086] Preparations, compositions, formulations, and products containing ammonia-oxidizing microorganisms This disclosure provides, in particular, compositions and preparations comprising ammonia-oxidizing microorganisms, such as purified and / or optimized preparations comprising AOM, formulations comprising AOM, and various products comprising AOM, such as natural products, non-natural products, enhanced natural products, consumer products, therapeutic products, or cosmetic products. The terms preparations, compositions, formulations, and products may be used interchangeably herein.

[0087] Any embodiment, preparation, composition, formulation, or product of ammonia-oxidizing microorganisms discussed herein may contain, be essentially, or be essentially (optionally in pure culture) ammonia-oxidizing microorganisms, such as live ammonia-oxidizing microorganisms.

[0088] The preparation may contain or be supplemented with products or by-products of ammonia-oxidizing microorganisms, such as nitrites, nitrates, nitric oxide, and CoQ8. In at least some embodiments, the preparation may contain or be supplemented with compositions that promote the growth or metabolism of ammonia-oxidizing microorganisms, promote the production of products or by-products of ammonia-oxidizing microorganisms, promote urease activity, or have a synergistic effect with ammonia-oxidizing microorganisms, such as ammonia, ammonium salts, urea, and urease. For example, the preparation may be supplemented with one or more of NO, nitrites, nitrates, CoQ8, ammonia, ammonium salts, urea, and urease. The dietary supplement may be included in the same formulation as the ammonia-oxidizing microorganisms, or in separate formulations for co-administration or combination administration. The dietary supplement formulation may be prepared to be delivered via any mode of delivery, such as inhalation of NO, nitrites, or nitrates. The preparation may contain, among other things, at least one of ammonia, ammonium salts, and urea. The preparation may contain or be supplemented with anti-inflammatory agents or compositions that produce anti-inflammatory effects.

[0089] This disclosure provides a preparation containing ammonia-oxidizing microorganisms for cosmetic use.

[0090] This disclosure provides preparations containing ammonia-oxidizing microorganisms for therapeutic use.

[0091] In some embodiments, the ammonia-oxidizing microorganism preparation may contain, for example, an effective amount of ammonia-oxidizing microorganisms in a concentration or quantity sufficient to have the desired cosmetic effect. The preparation may be formulated and / or delivered to deliver the desired cosmetic effect topically and / or systemically.

[0092] In some embodiments, the ammonia-oxidizing microorganism preparation may contain, for example, an effective amount of ammonia-oxidizing microorganisms in a concentration or quantity sufficient to have a desired therapeutic effect, for example, sufficient to treat at least partially a condition or disease. The preparation may be formulated and / or delivered to deliver the desired therapeutic effect topically and / or systemically.

[0093] In some embodiments, the ammonia-oxidizing microorganism preparation may contain a concentration or amount sufficient to alter, for example, a sufficient amount of ammonia-oxidizing microorganisms to reduce or increase the amount, concentration, or ratio of bacteria, or the genus of bacteria, in the subject. The bacteria may be non-pathogenic, pathogenic, or potentially pathogenic.

[0094] In some embodiments, the ammonia-oxidizing microorganism preparation may contain a concentration or amount sufficient to modulate the microbiome associated with the subject, for example, an effective amount of ammonia-oxidizing microorganisms.

[0095] In some embodiments, the ammonia-oxidizing microorganism preparation may contain a concentration or amount sufficient to deliver NO to a target, e.g., an effective amount of ammonia-oxidizing microorganisms. The ammonia-oxidizing microorganism preparation may contain a concentration or amount, e.g., an effective amount of ammonia-oxidizing microorganisms, such that when administered, the preparation modulates, alters, or changes the level of nitrite or NO in the target tissue or in circulation. For example, the ammonia-oxidizing microorganism preparation may contain a concentration or amount, e.g., an effective amount of ammonia-oxidizing microorganisms, such that when administered, the preparation increases the level of nitrite or NO in the target tissue or in circulation.

[0096] This disclosure provides, in particular, non-limiting compositions comprising ammonia-oxidizing microorganisms, such as N. eutropha, e.g., purified preparations of optimized N. eutropha. In some embodiments, the N. eutropha in the composition is optimized for growth rate, optimized for NH4 + Oxidation rate and optimization of NH4 + It possesses at least one characteristic selected from resistances.

[0097] In some embodiments, the Disclosure provides compositions having a specified number of species. The composition may contain only one type of species, for example, one type of ammonia-oxidizing microorganism. The Disclosure also provides compositions having, for example, N. eutropha and one other type of organism, but not the other type of organism. In other embodiments, the composition may have, for example, N. eutropha and 2, 3, 4, 5, 6, 7, 8, 9, or 10 other types of organism, but not the other type of organism. The other type of organism in this composition may be bacteria, such as ammonia-oxidizing bacteria, for example. Suitable ammonia-oxidizing microorganisms for this purpose include microorganisms of the genera Nitrosomonas, Nitrosococcus, Nitrosospira, Nitrosocystis, Nitrosolobus, or Nitrosovibrio. Similarly, the composition may also contain AOA.

[0098] In some embodiments, a composition containing, for example, N. eutropha, provides conditions that support the viability of N. eutropha. For example, the composition may promote the growth and metabolism of N. eutropha, or promote a dormant state (e.g., a frozen state) from which live N. eutropha can be recovered. If the composition promotes growth or metabolism, it may contain water and / or nutrients consumed by N. eutropha, such as ammonium, ammonia, urea, oxygen, carbon dioxide, or trace minerals. In some embodiments, a composition containing ammonia-oxidizing microorganisms provides conditions that support the viability of ammonia-oxidizing microorganisms. For example, the composition may promote the growth and metabolism of ammonia-oxidizing microorganisms, or promote a dormant state (e.g., a frozen state) or storage state as described herein from which live ammonia-oxidizing microorganisms can be recovered. If the composition promotes growth or metabolism, it may contain water and / or nutrients consumed by ammonia-oxidizing microorganisms, such as ammonium ions, ammonia, urea, oxygen, carbon dioxide, or trace minerals.

[0099] In some embodiments, one or more other organisms, such as organisms other than ammonia-oxidizing microorganisms, may be included in the ammonia-oxidizing microorganism preparation. For example, a group of organisms or genera selected from the group consisting of Lactobacillus, Streptococcus, Bifidobacter, and combinations thereof may be provided in the ammonia-oxidizing microorganism preparation. In some embodiments, the preparation may be substantially free of other organisms.

[0100] The preparation of ammonia-oxidizing microorganisms is approximately 10 3 From about 10 14 It may contain between CFU / ml. In some embodiments, the ammonia-oxidizing microorganism preparation contains at least about 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8, 10 9 , 10 10 , 10 11 , 2×10 11 , 5×10 11 , 10 12 , 2×10 12 , 5×10 12 , 10 13 , 2×10 13 , 5×10 13 , or 10 14 CFU / ml, or more, or about 10 3 ~10 4 , 10 4 ~10 5 , 10 6 ~10 7 , 10 7 ~10 8 , 10 8 ~10 9 , 10 9 ~10 10 , 10 10 ~10 11 , 10 11 ~10 12 , 10 12 ~10 13 , or 10 13 ~10 14 CFU / ml may be included.

[0101] In some embodiments, the preparation of ammonia-oxidizing microorganisms may contain between about 1×10 9 and about 10×10 9 CFU / ml. In some embodiments, the dosage of the preparation may be about 3×10 10 CFU per day, for example 3×10 10 CFU may be included. In some embodiments, the dosage of the preparation may be about 1×10 9 to about 10×10 9 CFU, for example, about 1×10 9 to about 10×10 9 CFU per day may be included. In some embodiments, the dosage of the preparation may be about 10 3 , 10 4 , 10 5 , 10 6 , 10 7, 10 8 , 10 9 , 10 10 , 10 11 , 2×10 11 , 5×10 11 , 10 12 , 2×10 12 , 5×10 12 , 10 13 , 2×10 13 , 5×10 13 , or 10 14 , or about 10 3 ~10 4 , 10 4 ~10 5 , 10 6 ~10 7 , 10 7 ~10 8 , 10 8 ~10 9 , 10 9 ~10 10 , 10 10 ~10 11 , 10 11 ~10 12 , 10 12 ~10 13 , or 10 13 ~10 14 It's fine to include CFU.

[0102] In some embodiments, the dosage of the preparation is at least about 7 × 10 per week. 10 CFU, for example, 21 x 10 10 It may contain CFU. In some embodiments, the dosage of the preparation is about 1 × 10⁶ per week. 9 From approximately 10 x 10 9 CFU, for example, approximately 1 x 10⁻⁶ per week 9 From approximately 10 x 10 9 It may contain CFU. In some embodiments, the dosage of the preparation is about 10 per week. 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 2×10 11, 5×10 11 , 10 12 , 2×10 12 , 5×10 12 , 10 13 , 2×10 13 , 5×10 13 , or 10 14 , or more, or about 10 3 ~10 4 , 10 4 ~10 5 , 10 6 ~10 7 , 10 7 ~10 8 , 10 8 ~10 9 , 10 9 ~10 10 , 10 10 ~10 11 , 10 11 ~10 12 , 10 12 ~10 13 , or 10 13 ~10 14 It's fine to include CFU.

[0103] In some embodiments, the dosage of the preparation is at least about 30 × 10 per month. 10 CFU, for example 90 x 10 10 It may contain CFU. In some embodiments, the dosage of the preparation is about 1 × 10 per month. 9 From approximately 10 x 10 9 CFU, for example, approximately 1 x 10⁻⁶ per month 9 From approximately 10 x 10 9 It may contain CFU. In some embodiments, the dosage of the preparation is about 10 per month. 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 2×10 11 , 5×10 11 , 10 12 , 2×10 12 , 5×10 12, 10 13 , 2×10 13 , 5×10 13 , or 10 14 , or more, or about 10 3 ~10 4 , 10 4 ~10 5 , 10 6 ~10 7 , 10 7 ~10 8 , 10 8 ~10 9 , 10 9 ~10 10 , 10 10 ~10 11 , 10 11 ~10 12 , 10 12 ~10 13 , or 10 13 ~10 14 It's fine to include CFU.

[0104] In some embodiments, the ammonia-oxidizing microorganism preparation may contain ammonia-oxidizing microorganisms in amounts ranging from about 0.1 milligrams (mg) to about 1000 mg. In certain embodiments, the preparation may contain ammonia-oxidizing microorganisms in amounts ranging from about 50 mg to about 1000 mg. The preparation may contain approximately 0.1-0.5 mg, 0.2-0.7 mg, 0.5-1.0 mg, 0.5-2 mg, 0.5-5 mg, 2.5-5 mg, 2.5-7.0 mg, 5.0-10 mg, 7.5-15 mg, 10-15 mg, 15-20 mg, 15-25 mg, 20-30 mg, 25-50 mg, 25-75 mg, 50-75 mg, 50-100 mg, 75-100 mg, 1 It may include amounts between 00-200mg, 200-300mg, 300-400mg, 400-500mg, 500-600mg, 600-700mg, 700-800mg, 800-900mg, 900-1000mg, 100-250mg, 250-500mg, 100-500mg, 500-750mg, 750-1000mg, or between 500-1000mg.

[0105] Advantageously, the formulation may have a pH level that promotes the viability, e.g., metabolic activity, of AOM, for example, N. eutropha. Urea is hydrolyzed to ammonia, raising the pH to 7 to 8. AOB is highly active in this pH range, and NH3 is converted to ammonium, lowering the pH to about 6, where it becomes unavailable. Lower pH levels, e.g., about pH 4, are also acceptable.

[0106] Ammonia-oxidizing microorganisms, such as N. eutropha, can be combined with one or more pharmaceutically or cosmetically acceptable additives. In some embodiments, “pharmaceutically acceptable additive” refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. In some embodiments, each additive is “pharmaceutically acceptable” in the sense that it is compatible with other components of a pharmaceutical formulation and is suitable for use in contact with human and animal tissues or organs in proportion to a reasonable benefit / risk ratio, without excessive toxicity, irritation, allergic reactions, immunogenicity, or other problems or complications. Remington: The Science and Practice of Pharmacy, 21st edition; Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients, 6th edition; Rowe et al. (eds.); The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd edition; Ash and Ash (eds.); Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd edition; Gibson (ed.); CRC Press LLC: Boca See Raton, Fla., 2009.

[0107] In some embodiments, cosmetically permissible additives represent cosmetically permissible materials, compositions, or vehicles, such as liquid or solid fillers, diluents, solvents, or encapsulating materials. In some embodiments, each additive is cosmetically permissible in the sense that it is compatible with other components of the cosmetic formulation and is suitable for use in contact with human and animal tissues or organs in proportion to a reasonable benefit / risk ratio, without excessive toxicity, irritation, allergic reactions, immunogenicity, or other problems or complications.

[0108] Although the active ingredient, for example, an ammonia-oxidizing microorganism, such as N. eutropha, can be administered alone, in many embodiments it is present in a pharmaceutical formulation or composition. Therefore, this disclosure provides a pharmaceutical formulation comprising an ammonia-oxidizing microorganism, such as N. eutropha, and a pharmaceutically acceptable additive. The pharmaceutical composition may take the following forms of pharmaceutical formulations.

[0109] According to one or more embodiments, preparations of ammonia-oxidizing microorganisms can be formulated to facilitate their desired delivery mechanism or mode of administration. The formulations described herein, for example, pharmaceutical or cosmetic formulations, include those suitable for oral administration, enteral administration (including oral, sublingual, sublabial, and rectal administration), parenteral administration (including subcutaneous, intradermal, intramuscular, intravenous, and intra-articular administration), inhalation (including various types of quantified doses, including particulate dust or mist that can be generated by pressurized aerosols, nebulizers, or inhalers, including intranasal or pulmonary administration), intranasal administration, ocular administration, ocular administration, rectal administration, injection, genitourinary administration, and topical administration (including cutaneous, transdermal, transmucosal, oral, sublingual, and intraocular administration), but the most appropriate route may depend, for example, on the recipient's condition or impairment.

[0110] According to one or more non-limiting embodiments, preparations containing ammonia-oxidizing microorganisms can be administered to a subject, for example, for cosmetic or therapeutic purposes, as a solution, suspension, powder, liquid, drop, spray, aerosol, mist, emulsion, foam, cream, ointment, gel, hydrogel, resin, tablet, capsule, film, suppository, enema, douche, pessary, insert, patch, such as a transdermal patch, or implantable device, such as a stent, catheter, vaginal ring, or intrauterine device.

[0111] Also disclosed are devices configured to deliver preparations containing live ammonia-oxidizing microorganisms via a desired mode of administration or via targeted delivery.

[0112] According to one or more embodiments, a preparation can be formulated for targeted delivery to a subject, for example, to a target tissue, target region, target system, or target organ of the subject. For example, a preparation can be formulated for delivery to the eyes, ears, nose, genitourinary system, respiratory system, or digestive system of the subject. In some embodiments, targeted delivery may be based on the condition or disorder of the subject. For example, a preparation for targeted delivery may be based on a desired local or systemic effect to be achieved, for example, a local or systemic therapeutic effect or cosmetic effect. In some embodiments, the target tissue, target region, target system, or target organ of the subject may be selected for its relevance to the desired local or systemic effect.

[0113] The formulations can be conveniently presented as unit dosage forms and can be prepared by any method known in the pharmaceutical art. Typically, the method involves associating an active ingredient (e.g., an ammonia-oxidizing microorganism, e.g., N. eutropha) with a pharmaceutical carrier containing one or more auxiliary components. Generally, the formulations are prepared by uniformly and closely associating the active ingredient with a liquid carrier, a micronized solid carrier, or both, and then, if necessary, shaping the product into the desired formulation.

[0114] Formulations may be presented as separate units, such as capsules, cachets, or tablets, each containing a predetermined amount of, for example, N. eutropha; as powders or granules; as solutions or suspensions in aqueous or achromatic liquids; or as oil-in-water or water-in-oil liquid emulsions. Formulations, such as solutions, aerosols, sprays, and mists, may be in multi-dose forms, for example, packaged units containing a predetermined number of doses or single doses, for example, packaged units containing a single dose. The active ingredient may also be presented as a bolus, lick, or paste. Various pharmaceutically acceptable carriers and their formulations are described in standard pharmaceutical textbooks, e.g., Remington's Pharmaceutical Sciences by E.W. Martin. See also Wang, Y.J. and Hanson, MA, Journal of Parenteral Science and Technology, Technical Report 10, Supplement 42:2S, 1988.

[0115] Ammonia-oxidizing microorganisms, such as N. eutropha compositions, can be administered in forms suitable for immediate or extended release. Suitable examples of sustained-release systems include suitable polymer materials, such as semipermeable polymer matrices in the form of molded articles, e.g., films or microcapsules; suitable hydrophobic materials, e.g., as emulsions in acceptable oils; or ion exchange resins. Sustained-release systems can be administered orally; rectally; parenterally; intracapsule; intravaginally; intraperitoneally; topically, e.g., as powders, ointments, gels, drops or transdermal patches; buccally; or as sprays.

[0116] The preparations for administration can be appropriately formulated to provide controlled release of ammonia-oxidizing microorganisms, such as N. eutropha. For example, the pharmaceutical composition may be in the form of particles containing one or more of the following: biodegradable polymers, polysaccharide gels and / or bioadhesive polymers, or amphiphilic polymers. These compositions exhibit certain biocompatibility features that enable controlled release of the active substance. See U.S. Patent No. 5,700,486.

[0117] Exemplary compositions may include, for example, a suspension containing crystalline cellulose for bulking, alginic acid or sodium alginate as a suspending agent, methylcellulose, dicalcium phosphate, starch, magnesium stearate and / or lactose and / or other additives, binders, fillers, disintegrants, diluents and lubricants, mannitol, lactose, sucrose and / or cyclodextrin. Such formulations may also include high molecular weight additives such as cellulose (Avicel) or polyethylene glycol (PEG). Such formulations may also include additives to aid mucosal adhesion, such as hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), sodium carboxymethylcellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and agents to control release, such as polyacrylic acid copolymer (e.g., Carbopol 934). Lubricants, flow enhancers, fragrances, colorants and stabilizers may be added to facilitate preparation and use. The surfactant may be a zwitterionic surfactant, a nonionic surfactant, or an anionic surfactant.

[0118] Additives such as surfactants that may be used in embodiments of this disclosure include cocamidopropyl betaine (ColaTeric COAB), polyethylene sorbitol ester (e.g., Tween 80), ethoxylated lauryl alcohol (RhodaSurf 6 NAT), sodium laureth sulfate / lauryl glucoside / cocamidopropyl betaine (Plantapon 611 L UP), sodium laureth sulfate (e.g., RhodaPex ESB 70 NAT), and alkyl polyglucosides (e.g., Plantaren). Dr. Bronner's Castile Soap (2000 N UP), sodium laureth sulfate (Plantaren 200), Dr. Bronner's Castile Soap, Dr. Bronner's Castile Baby Soap, lauramine oxide (ColaLux Lo), sodium dodecyl sulfate (SDS), alkyl polyglucoside polysulfonate (PolySufanate 160 P), sodium lauryl sulfate (Stepanol-WA Extra K), and combinations thereof may contain one or more of these. Dr. Bronner's Castile Soap and Dr. Bronner's Baby Soap contain water, organic coconut oil, potassium hydroxide, organic olive oil, fair trade organic hemp oil, organic jojoba oil, citric acid, and tocopherol.

[0119] In some embodiments, the surfactant may be used in combination with ammonia-oxidizing microorganisms in an amount that causes nitrite formation. In some embodiments, the preparation may contain surfactant ranging from less than about 0.0001% to about 10%. In some embodiments, the preparation may contain surfactant between about 0.1% and about 10%. In some embodiments, the concentration of surfactant used may be between about 0.0001% and about 10%. In some embodiments, the preparation may be substantially surfactant-free.

[0120] In some embodiments, the formulation, for example, the preparation, may contain other components that enhance the effectiveness of ammonia-oxidizing microorganisms or their delivery, or enhance the treatment or indication.

[0121] In some embodiments, a chelating agent may be included in the preparation. The chelating agent may be another compound, for example, a compound that can bind to a metal. The chelating agent may help remove an undesirable compound from the environment, or it may act in a protective manner to reduce or eliminate contact between certain compounds and the environment, for example, ammonia-oxidizing microorganisms, for example, preparations of ammonia-oxidizing microorganisms, for example, additives. In some embodiments, the preparation may be substantially free of a chelating agent.

[0122] The formulations may also contain aqueous and non-aqueous sterile suspensions which may include antioxidants, buffers, bacteriostatic agents that prevent the growth of undesirable microorganisms, solutes, and suspending and thickening agents. The formulations may be presented in unit dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried state requiring only the addition of a sterile liquid carrier, such as saline or water for injection, immediately before use. Immediate solutions and suspensions may be prepared from the aforementioned types of powders, granules, and tablets. Exemplary compositions include, for example, a suitable non-toxic and pharmaceutically acceptable diluent or solvent such as mannitol, 1,3-butanediol, water, Ringer's solution, and isotonic sodium chloride solution, or other suitable dispersants or wetting and suspending agents including synthetic monoglycerides or diglycerides, and a solution or suspension which may contain fatty acids including oleic acid or cremaphor. The aqueous carrier may be an isotonic buffer solution with a pH of approximately 3.0 to 8.0, approximately 3.5 to 7.4, for example, 3.5 to 6.0, or for example, 3.5 to 5.0. Useful buffers include sodium citrate-citric acid and sodium phosphate-phosphate, and sodium acetate / acetic acid buffer. In some embodiments, the composition does not contain an oxidizing agent.

[0123] Possible additives include, for example, proteins such as human serum albumin or plasma preparations. If desired, the pharmaceutical composition may also contain small amounts of non-toxic auxiliary substances such as humectants or emulsifiers, preservatives, and pH buffers, e.g., sodium acetate or sorbitan monolaurate. In some embodiments, additives, e.g., pharmaceutically acceptable or cosmetically acceptable additives, may include anti-adhesives, binders, coatings, disintegrants, fillers, fragrances, colorants, lubricants, flow enhancers, adsorbents, preservatives, or sweeteners. In some embodiments, the preparation may be substantially free of additives.

[0124] In some embodiments, the preparation does not need to substantially contain one or more of the compounds or substances listed in this disclosure.

[0125] Exemplary compositions for spray, aerosol, or mist administration include, for example, a solution in saline solution that may contain benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, and / or other solubilizers or dispersants. Conveniently, in compositions for aerosol administration, the ammonia-oxidizing microorganism, e.g., N. eutropha, is delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, nitrogen, or other suitable gas. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve for delivering a fixed amount. For example, gelatin capsules and cartridges can be formulated to contain a powder mixture of N. eutropha and a suitable powder base, e.g., lactose or starch. In certain embodiments, N. eutropha is administered as an aerosol from a metering valve via an aerosol adapter, also known as an actuator. Optionally, the formulation may also include a stabilizer and / or porous particles for deep lung delivery (see, for example, U.S. Patent No. 6,447,743).

[0126] The formulation may be presented using a carrier such as cocoa butter, synthetic glyceride esters, or polyethylene glycol. Such carriers are typically solid at room temperature but liquefy and / or dissolve at body temperature to release ammonia-oxidizing bacteria, such as N. eutropha.

[0127] Exemplary topical administration compositions include a topical carrier such as Plastibase (mineral oil gelled with polyethylene). In some embodiments, the composition and / or additives may be in one or more forms of liquid, solid, or gel. For example, the suspension may, non-limitingly, include water, saline, phosphate-buffered saline, or ammonia-oxidized storage buffer. The gel formulation may, without limitation, contain agar, silica, polyacrylic acid (e.g., Carbopol®), carboxymethylcellulose, starch, guar gum, alginate, or chitosan. In some embodiments, the formulation may be supplemented with an ammonia source, not limited to ammonium chloride or ammonium sulfate.

[0128] In some embodiments, ammonia-oxidizing microorganisms, such as N. eutropha compositions, are formulated to improve the penetration of NO into, for example, the skin or other target tissues. Gel-forming materials, such as KY jelly or various hair gels, improve NO absorption in the skin by presenting a diffusion barrier against NO loss into the surrounding air. NO levels in the skin generally do not exceed 20 nM / L because that level activates GC, causing local vasodilation and oxidative breakdown of excess NO.

[0129] In particular, it should be understood that, in addition to the components mentioned above, the formulations described herein may also contain other conventional agents in the art with respect to the type of formulation in question.

[0130] Formulations, e.g., preparations, e.g., compositions, may be delivered in a container, delivery system, or delivery device having a weight that may be less than about 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500, or 2000 grams, with or without the contents of the container.

[0131] A suitable unit-dose formulation is a unit-dose formulation containing the effective dose or a suitable fraction of an ammonia-oxidizing microorganism, such as N. eutropha, as specified herein.

[0132] A therapeutically effective dose of an ammonia-oxidizing microorganism, such as N. eutropha, may be administered as a single pulse dose, a bolus dose, or as pulse doses administered over time. Thus, in pulse doses, a bolus dose of the ammonia-oxidizing microorganism, e.g., N. eutropha, is provided, followed by a period of administration of the microorganism to the subject, and then a second bolus dose. In specific and non-limiting examples, pulse doses may be administered during a 1-day cycle, a 1-week cycle, or a 1-month cycle.

[0133] In some embodiments, a preparation of ammonia-oxidizing microorganisms, e.g., a formulation, e.g., a composition, can be applied for a predetermined number of days. This may be based, for example, at least in part, on the severity of the condition or disease, the response to the treatment, the amount of medication applied, and the dose frequency. For example, the preparation may be applied for about 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, 84-91 days, about 1 month, about 2 months, or about 3 months. In some embodiments, the ammonia-oxidizing bacteria may be administered for an indefinite period, e.g., longer than 1 year, longer than 5 years, longer than 10 years, longer than 15 years, longer than 30 years, longer than 50 years, or longer than 75 years. In a particular embodiment, the preparation may be applied for approximately 16 days.

[0134] In some embodiments, a preparation of ammonia-oxidizing microorganisms, e.g., a formulation, e.g., a composition, may be applied a predetermined number of times per day. This may be based, for example, at least in part, on the severity of the condition or disease, the response to treatment, the amount of medication applied, and the dose frequency. For example, the preparation may be applied 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 times per day.

[0135] In some embodiments, the preparation may be applied once per day. In other embodiments, the preparation may be applied twice per day. In some embodiments, the preparation may be applied in a first predetermined amount for a certain number of days, and in a second predetermined amount for a certain number of subsequent days. In some embodiments, the preparation may be applied for about 16 days.

[0136] According to one or more embodiments, the preparation can typically be adapted to the physiological environment of the subject. In at least some embodiments, the composition is formulated to have a substantially neutral pH or a physiological pH, for example, the pH that is usually dominant at the target site, for intended delivery, administration, or desired effect. The composition can be formulated to have a pH between about 5.5 and about 8.5. The composition can be formulated to include conditions adapted to the target site of the physiological environment of the subject, such as pH and tonicity.

[0137] The preparation can be formulated for transmucosal delivery and / or circulation, for example, topically or systemically. In some embodiments, the preparation can be formulated so that at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of ammonia-oxidizing microorganisms, their products, or their by-products (e.g., nitrates, nitrites, NO, or CoQ8) penetrates into the deposited or target tissue. The preparation can be formulated so that 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of ammonia-oxidizing microorganisms, their products, or their by-products penetrates into the deposited or target tissue or enters circulation.

[0138] According to one or more embodiments, the preparation may be in the form of a solution, suspension, emulsion, cream, ointment, gel, hydrogel, or liquid for administration to a subject, such as a drop, spray, aerosol, or mist, tablet, capsule, or device.

[0139] According to one or more embodiments, preparations, compositions, formulations, or products containing ammonia-oxidizing microorganisms may undergo quality control and / or quality testing while they are being prepared and / or after their preparation is completed. International (PCT) Patent Application Publication WO2015 / 179669 (International (PCT) Patent Application PCT / US2015 / 032017, filed on 21 May 2015), which is incorporated herein by whole reference for all purposes, describes various methods for preparing materials having ammonia-oxidizing microorganisms and various methods for testing such materials. For example, preparations containing ammonia-oxidizing microorganisms can be assessed or evaluated by comparing one or more parameters, such as OD level, pH level, waste level, nutrient level, contaminant level, oxidation rate, nitrite level, and protein concentration, to predetermined values.

[0140] This disclosure provides, in particular, a kit comprising preparations of ammonia-oxidizing microorganisms disclosed herein. The formulations may comprise individual units of ammonia-oxidizing microorganisms, e.g., solid formulations, liquid formulations, or gaseous formulations. The formulations, e.g., solutions, aerosols, sprays, and mists, may be multi-dose formulations (multiple uses), e.g., packaged units comprising a predetermined number of doses, or single-dose formulations (single use), e.g., packaged units comprising a single dose. Preparations of ammonia-oxidizing microorganisms may be packaged in devices or containers configured to hold volumes of at least less than about 1 ml, 1 ml, 5 ml, 10 ml, 20 ml, 25 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml, 100 ml, or more than about 100 ml.

[0141] The kit may further include one or more devices for administering the preparation, such as a syringe, needle, catheter, enema, valve, pipette (eye dropper or ear dropper), and other drug administration devices known in the art. The kit may include instructions for use, such as instructions for administering the ammonia-oxidizing microorganisms disclosed herein, or instructions for combination therapies including the administration of ammonia-oxidizing microorganisms. The kit may include a second or subsequent composition for administration together with the ammonia-oxidizing preparations disclosed herein. For example, the kit may include a nutritional supplement or composition containing ammonia-oxidizing microorganism products or by-products, a composition that promotes the growth or metabolism of ammonia-oxidizing microorganisms, a composition that promotes the production of ammonia-oxidizing microorganism products or by-products, a composition that promotes urease activity, or a composition that has a synergistic effect with ammonia-oxidizing microorganisms, or a composition or pharmaceutical agent that treats related diseases, disorders, or symptoms of related diseases or disorders, such as an anti-inflammatory composition, which is approved to treat or commonly used to treat. The kit may include products that are “biome-friendly” or “biome-compatible” as disclosed herein, for example, one or more cosmetic products compatible with the microbiome. Any of the products included in the kit can be specifically formulated to treat a targeted indication and / or for a desired mode of delivery as described herein.

[0142] natural products, consumer products In some specific embodiments, the preparations containing ammonia-oxidizing microorganisms discussed herein may be natural products or consumer products. In other embodiments, the preparations containing ammonia-oxidizing microorganisms may be used together with natural products or consumer products instead.

[0143] Ammonia-oxidizing microorganisms, such as N. eutropha, may be associated with various natural products, examples of which are described below. These natural products may consist of formulations, compositions, or preparations disclosed throughout this disclosure.

[0144] Natural products may be or include commercial products, and may refer to cosmetics, dietary supplements, and foods produced from natural sources, such as foods, dietary supplements, medicinal foods, food additives, functional foods, or beverages. Natural products may have pharmacological or biological activity that may be therapeutically beneficial, for example, in the treatment of a disease or condition. Natural products may be included in traditional medicines, cosmetic treatments, and spa treatments. Natural products as referred to herein may include one or more of the components described as natural products incorporated into preparations or formulations containing additives, for example, one or more other components. Preparations or formulations referred to as natural products may include natural products as defined herein and one or more additional components or ingredients. Any of the compositions, preparations, or formulations discussed throughout this disclosure may be or include one or more natural products.

[0145] In some embodiments, the natural product or enhanced natural product may comprise at least one of mud, water, food-derived products, plant-derived products, extracts, and oils. The natural product or enhanced natural product may be used in spa treatments. In some embodiments, the natural product or enhanced natural product may be incorporated into at least one of powders, creams, lotions, wraps, scrubs, eye masks, facial masks, body masks, aerosols, such as mists, sprays, ointments, wipes, sticks, bandages, or immersions.

[0146] In some embodiments, natural or enhanced natural products include: baby products, e.g., baby shampoo, baby lotion, baby oil, baby powder, baby cream; bath preparations, e.g., bath oil, tablets, salt, bubble bath, bath capsules; eye makeup preparations, e.g., eyebrow pencil, eyeliner, eyeshadow, eye lotion, eye makeup remover, mascara; fragrance preparations, e.g., cologne, eau de toilette, perfume, powder (dusting and talcum), sachets; hair preparations, e.g., hair conditioner, hair spray, hair straightener, permanent waver, rinse, shampoo, tonic, hair styling products, styling aids, wave setters; hair coloring preparations, e.g., hair dyes and hair colors, hair dyes, coloring hair rinses, coloring hair shampoos, coloring hair lighteners, hair bleach; makeup preparations, e.g., face powder, foundation, leg and body paint, lipstick, etc. Makeup bases, blushes, and makeup fixatives; manicure preparations, e.g., base coats and undercoats, cuticle softeners, nail creams and lotions, nail extenders, nail polishes and enamels, nail polishes and enamel removers; oral hygiene products, e.g., toothpastes, mouthwashes and breath fresheners; bath soaps and detergents, deodorants, bidets, women's hygiene deodorants; shaving preparations, e.g., aftershave lotions, beard softeners, talcum, pre-shave lotions, shaving creams, and shaving soaps; skincare preparations, e.g., cleansers, depilators, powders and sprays for face and neck, body and hands, and feet, moisturizers, night preparations, paste masks, and skin fresheners; and tanning preparations, e.g., gels, creams, and liquids, and indoor tanning preparations, which may be provided as or contained within at least one of these.

[0147] Ammonia-oxidizing bacteria, such as N. eutropha, may be associated with a variety of consumer products, examples of which are described later, but may also consist of formulations, compositions, or preparations disclosed throughout this disclosure. In some embodiments, the ammonia-oxidizing bacteria, such as N. eutropha, associated with a product are mixed with the product, for example, spread uniformly throughout the product, and in some embodiments, the ammonia-oxidizing bacteria, such as N. eutropha, associated with a product are layered on top of the product.

[0148] In some embodiments, the preparation may be in or provided as a powder, cosmetic, cream, stick, aerosol, such as mist, ointment, wipe, or bandage.

[0149] In some embodiments, ammonia-oxidizing bacteria, such as N. eutropha, are associated with the powder. The powder is typically a small, particulate solid that does not adhere to one another and can flow freely when tilted. Exemplary powders for consumer use include talcum powder and certain cosmetics (e.g., powder foundations).

[0150] In some embodiments, ammonia-oxidizing bacteria are associated with cosmetics. Cosmetics may be topical application substances intended to alter human appearance, such as liquid foundation, powder foundation, rouge, or lipstick, and may be referred to as preparations. Cosmetics may be any substance listed under the regulations of the Food and Drug Administration, for example, in 21 § 720.4 of the Code of Federal Regulations.

[0151] In some embodiments, ammonia-oxidizing microorganisms, such as N. eutropha, are associated with the cosmetic. The cosmetic may be a topical application substance intended to alter a person's appearance, such as a liquid foundation, powder foundation, rouge, or lipstick. Other components may be added to pharmaceutical formulations, such as preparations, or cosmetic preparations, as selected by those skilled in the art, including water, mineral oil, colorants, perfumes, aloe, glycerin, sodium chloride, sodium bicarbonate, pH buffers, UV blockers, silicone oils, natural oils, vitamin E, herbal concentrates, lactic acid, citric acid, talc, clay, calcium carbonate, magnesium carbonate, zinc oxide, starch, urea, and erythorbic acid, or any other additives known to those skilled in the art, including the additives disclosed herein.

[0152] Preparations, for example, cosmetics include baby products such as baby shampoo, baby lotion, baby oil, baby powder, and baby cream; bath preparations such as bath oil, tablets, salt, bubble bath, and bath capsules; eye makeup preparations such as eyebrow pencils, eyeliners, eyeshadows, eye lotions, eye makeup removers, and mascaras; fragrance preparations such as colognes, eau de toilettes, perfumes, powders (dusting and talcum), and sachets; hair preparations such as hair conditioners, hair sprays, hair straighteners, permanent wavers, rinses, shampoos, tonics, hair styling products, styling aids, and wave setters; hair coloring preparations such as hair dyes and hair colors, hair dyes, coloring hair rinses, coloring hair shampoos, coloring hair lighteners, and hair bleaches; and makeup preparations such as face powders, foundations, leg and body paints, and lipsticks. Makeup bases, blushes, makeup fixatives; manicure preparations, e.g., base coats and undercoats, cuticle softeners, nail creams and lotions, nail extenders, nail polishes and enamels, nail polishes and enamel removers; oral hygiene products, e.g., toothpastes, mouthwashes and breath fresheners; bath soaps and detergents, deodorants, bidets, women's hygiene deodorants; shaving preparations, e.g., aftershave lotions, beard softeners, talcum, pre-shave lotions, shaving creams, shaving soaps; skincare preparations, e.g., cleansers, depilators, powders and sprays for face and neck, body and hands, and feet, moisturizers, night preparations, paste masks, skin fresheners; and tanning preparations, e.g., gels, creams, and liquids, and indoor tanning preparations, which may be at least one of these.

[0153] In some embodiments, the formulations, compositions, or preparations described herein may be provided as or contained therein, comprising at least one of the following: baby products, e.g., baby shampoo, baby lotion, baby oil, baby powder, baby cream; bath preparations, e.g., bath oil, tablets, salt, bubble bath, bath capsule; powders (dusting and talcum), sachets; hair preparations, e.g., hair conditioner, rinse, shampoo, tonic, face powder, cuticle softener, nail cream and lotion; oral hygiene products, mouthwash, bath soap, bidet, women's hygiene deodorant; shaving preparations, e.g., aftershave lotion; skincare preparations, e.g., cleansing, powder and spray for face and neck, body and hands, and feet; moisturizers; night preparations, paste masks, skin fresheners; and sunscreen preparations, e.g., gels, creams, and liquids.

[0154] In some embodiments, ammonia-oxidizing microorganisms, such as N. eutropha, are associated with aerosols, sprays, or mists, and these terms can be used interchangeably. An aerosol is typically a colloid of fine solid particles or fine droplets in a gas, such as air. Aerosols can be created by placing N. eutropha (and optionally a carrier) in a pressurized vessel and then opening a valve to release the contents. The vessel can be designed to apply only a pressure level suitable for the viability and suitability of N. eutropha. For example, high pressure may be applied only for a short time, and / or the pressure may be low enough not to impair viability. Examples of consumer use of aerosols include sunscreens, deodorants, perfumes, hairsprays, and insect repellents. Aerosols may also be referred to as sprays or mists.

[0155] A composition containing an ammonia-oxidizing microorganism, such as N. eutropha, may also contain one or more of the following: a humectant, a deodorant, a fragrance, a colorant, an insect repellent, a cleansing agent, or a UV blocker.

[0156] In some embodiments, ammonia-oxidizing microorganisms, such as N. eutropha, are associated with fabrics, yarns, or threads. Clothing items such as shoes, shoe insoles, pajamas, sneakers, belts, hats, shirts, underwear, sportswear, helmets, towels, gloves, socks, bandages, and similar items may also be treated with ammonia-oxidizing bacteria, such as N. eutropha. Bedding, including sheets, pillows, pillowcases, and blankets, may also be treated with ammonia-oxidizing bacteria, such as N. eutropha. In some embodiments, skin areas that cannot be washed for a certain period of time may also come into contact with ammonia-oxidizing bacteria, such as N. eutropha. For example, skin surrounded by an orthopedic cast that immobilizes an injured limb during the healing process, and areas near injuries that must remain dry for proper healing, such as sutured wounds, may benefit from contact with ammonia-oxidizing bacteria, such as N. eutropha.

[0157] In some embodiments, this disclosure provides wearables containing the ammonia-oxidizing microorganisms described herein. Wearables may be lightweight and can be closely attached to the wearer's body in a manner that does not interfere with walking. Examples of wearables include watches, wristbands, headbands, hair ties, hair nets, shower caps, hats, hairpieces, and jewelry. Wearables containing the ammonia-oxidizing bacteria described herein, for example, strain N. eutropha, may be provided in concentrations that provide, for example, one or more of the following: treatment or prevention of skin disorders, treatment or prevention of diseases or conditions associated with low nitrite levels, treatment or prevention of body odor, treatment to supply nitric oxide to a subject, or treatment to inhibit the growth of microorganisms.

[0158] In some embodiments, ammonia-oxidizing microorganisms, such as N. eutropha, are associated with products intended to come into contact with hair, such as brushes, combs, shampoos, conditioners, headbands, hair ties, hair nets, shower caps, hats, and hairpieces. Nitric oxide formed on the hair away from the skin surface may be trapped inside a hat, scarf, or face mask and directed to inhaled air.

[0159] Items that come into contact with human surfaces, such as diapers, may be accompanied by ammonia-oxidizing microorganisms, such as N. eutropha. Since diapers are designed to hold and contain urine and feces produced by incontinent individuals, urea in urine and feces can be hydrolyzed by skin and fecal bacteria to form free ammonia, which is irritating and can cause diaper rash. Incorporating ammonia-oxidizing bacteria, such as N. eutropha, that metabolize urea into nitrite or nitrate can avoid the release of free ammonia and may instead release nitrite and ultimately NO, which may help maintain healthy skin for both children and incontinent adults. The release of nitric oxide in diapers may also have an antimicrobiological effect against disease-causing organisms present in human feces. This effect can persist even after disposable diapers are discarded as waste, potentially reducing the incidence of disease transmission through contact with soiled disposable diapers.

[0160] In some embodiments, products containing ammonia-oxidizing microorganisms, such as N. eutropha, are packaged. The packaging may serve to compress the product or protect it from damage, contamination, or deterioration. The packaging may include, for example, plastic, paper, cardboard, or wood. In some embodiments, the packaging is bacterial-impermeable. In some embodiments, the packaging is oxygen and / or carbon dioxide-permeable.

[0161] Treatment methods using ammonia-oxidizing microorganisms According to one or more embodiments, a subject may be treated by administering ammonia-oxidizing microorganisms, for example, a preparation containing ammonia-oxidizing microorganisms. When used herein, treatment of a subject may involve administering an ammonia-oxidizing microorganism composition for cosmetic or therapeutic results. For example, treatment may involve treating or alleviating a condition, symptom, or condition-related side effect, or achieving a desired cosmetic effect.

[0162] Subjects may include animals, mammals, humans, non-human animals, domesticated animals, or companion animals. Subjects may be female or male. Subjects may have a variety of skin types. Subjects may have a variety of health-related profiles, including medical history and / or genetic predispositions. Subjects may typically have a normal microbiome, e.g., a physiological microbiome, or a disturbed microbiome. Subjects may be characterized as one of the following ethnic / racial groups: Asian, Black or African American, Hispanic or Latino, White, or multi-ethnic. Subjects may be under 1 year old, or between 1 and 5 years old, between 5 and 10 years old, between 10 and 20 years old, between 20 and 30 years old, between 30 and 40 years old, between 40 and 50 years old, between 50 and 60 years old, or over 60 years old.

[0163] Ammonia-oxidizing microorganisms that can be used to treat the target include all ammonia-oxidizing microorganisms, for example, the N. eutropha compositions described herein, for example, optimized ammonia-oxidizing microorganisms, for example, purified preparations of strain D23.

[0164] Methods for administering or delivering therapeutic or cosmetic products may be provided. These methods may involve administering or introducing a preparation containing live ammonia-oxidizing microorganisms to a target. The preparation can be formulated to treat a target indication and / or for a desired mode of delivery.

[0165] According to one or more embodiments, a preparation containing live ammonia-oxidizing microorganisms can be administered to a first tissue of interest. The first tissue may be a depositional tissue. The first tissue may be a target tissue or a tissue other than a target tissue. The live ammonia-oxidizing microorganisms, or their products, such as nitrite and / or nitric oxide, can then be moved or transported to a second tissue, for example, by diffusion. The second tissue may be a target tissue. The target tissue may be related to a desired local or systemic effect. The target tissue may be related to an indication, disorder, or condition to be treated.

[0166] Ammonia-oxidizing microbial preparations can be administered, for example, to the skin for cosmetic or therapeutic effects. For example, the administration may provide a cosmetic treatment, benefit, or effect. In some embodiments, the administration may provide one or more treatments or improvements to oily appearance, pore appearance, shine, blemishes, skin tone uniformity, visual smoothness, and tactile smoothness. In some embodiments, the cosmetic appearance of the subject may be altered, for example, resulting in improved skin health. Signs of aging may be reduced, delayed, or reversed. The administration may qualitatively improve the condition and / or quality of the skin and / or scalp. Skin smoothness, hydration, firmness, and / or softness in the subject may be improved. This disclosure also provides a method for reducing body odor.

[0167] The administration may provide a therapeutic treatment, benefit, or effect. This disclosure provides methods for supplying nitrite and nitric oxide to a target. This disclosure provides various methods for using ammonia-oxidizing microorganisms to suppress, treat, or prevent diseases, disorders, infections, and conditions. Ammonia-oxidizing microorganisms may be used, for example, to treat various diseases associated with low nitrite levels, skin diseases, and diseases caused by pathogenic bacteria. In some embodiments, the administration may provide a reduction in inflammation. In fact, topical or systemic anti-inflammatory effects may be demonstrated. In some non-limiting embodiments, inflammation may be downregulated. In at least some embodiments, the growth of microorganisms may be inhibited. Skin and overall health may be improved. Inadequate circulation may be enhanced. Endothelial function may be promoted. Changes in the levels of nitrite or NO in or in circulation in target tissues may be demonstrated. In some embodiments, the administration, e.g., an effective dose, may modulate, alter, or change the levels of nitrite or NO in or in circulation in target tissues. In some embodiments, administration, for example, an effective dose, may increase the levels of nitrite or NO in or in the target tissue.

[0168] Administration of the compositions disclosed herein may provide transmucosal delivery and / or circulation, for example, topically or systemically. In some embodiments, administration may provide penetration of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of ammonia-oxidizing microorganisms, their products, or their by-products (e.g., nitrates, nitrites, NO, or CoQ8) into deposited or target tissue. In at least some embodiments, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of ammonia-oxidizing microorganisms, their products, or their by-products penetrate into deposited or target tissue or enter circulation upon administration of the compositions disclosed herein.

[0169] The preparations and methods of this disclosure may provide a way to reduce the amount of undesirable microorganisms from a given environment. The ammonia-oxidizing microorganisms described herein can eliminate other organisms, for example, by consuming poor nutrient levels or generating byproducts harmful to other organisms, or by altering the pH level to a level that does not aid in the growth of undesirable organisms.

[0170] This disclosure also provides a method for promoting wound healing, including chronic wounds, in patients with impaired healing capacity, such as diabetic patients. A bandage containing ammonia-oxidizing microorganisms may optionally be applied to the wound.

[0171] It is understood that many modern degenerative diseases may be caused by a deficiency of NO species, and that AOM can be administered to supply these species to target tissues directly or via diffusion. The application of AOM can resolve long-term medical conditions. In certain embodiments, AOM is applied to subjects, in particular, to compensate for modern bathing habits using anionic detergents that remove AOM from the surface skin.

[0172] According to one or more embodiments, AOM converts ammonia into nitrites, antimicrobiome compounds, and nitric oxide, which are signaling molecules in inflammatory processes well described in the literature.

[0173] This disclosure provides, in particular, a method for modulating the composition of a microbiome, for example, a method for modulating or altering the ratio of microbiomes in an environment, for example, a surface, for example, a surface of an object. This may, by extension, demonstrate health-related benefits. The method may include administering a preparation containing ammonia-oxidizing microorganisms to an object. In some embodiments, the amount and frequency of administration, for example, application, may be sufficient to reduce the ratio of pathogenic microorganisms.

[0174] The application of ammonia-oxidizing microorganisms to a target population, such as humans, can lead to unexpected changes in the microbiome. This could result in an increase in the proportion of normal, commensal, non-pathogenic species and a decrease in the proportion of potentially pathogenic, pathogenic, or disease-causing organisms.

[0175] An increase in the proportion of non-pathogenic bacteria may occur over predetermined periods, such as 1 day, 2 days, 3 days, 4 days, 5 days, 1 week, 2 weeks, 3 weeks, or less than 4 weeks, or 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, or 84-91 days.

[0176] A decrease in the proportion of pathogenic bacteria may occur over predetermined periods, such as less than 1 day, 2 days, 3 days, 4 days, 5 days, 1 week, 2 weeks, 3 weeks, or 4 weeks, or less than 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, or 84-91 days.

[0177] According to one or more embodiments, subjects may be evaluated for the need for treatment. In some embodiments, subjects may be selected based on the subjects requiring treatment. The disclosure may further provide obtaining samples from subjects and analyzing the samples. In some embodiments, subjects may be evaluated, for example, at predetermined time intervals before, during, and / or after treatment.

[0178] According to one or more embodiments, administration can be carried out before, during, or after the onset of a health-related condition, or in response to a prodromal, trigger, or symptom thereof. According to one or more embodiments, a second amount, for example, a second dose preparation, can be administered to the subject.

[0179] In certain embodiments, the Disclosure provides a combination therapy comprising an ammonia-oxidizing microorganism, e.g., N. eutropha, and a second treatment, e.g., a second therapeutic agent. For example, the Disclosure provides a physical mixture in which two (or more) therapies are physically mixed. In other embodiments, the two (or more) therapies are administered in combination as separate formulations. The second therapy may be, for example, a pharmaceutical agent, surgical procedure, or any other medical approach that is approved to treat or commonly used to treat, an associated disease, disorder, or symptoms of an associated disease or disorder. The second treatment may be administered before or after administration. An effective dose may be administered concurrently with the second treatment. The second treatment may be administered via the same or different modes of delivery. The subject may have a therapeutic level of the second treatment at the time of administration of the preparation. In certain embodiments, the second treatment may be administered to produce an anti-inflammatory effect or to reduce inflammation at a target site. In at least some embodiments, the preparation may be administered simultaneously with or together with products or by-products of ammonia-oxidizing microorganisms, such as nitrites, nitrates, nitric oxide, and CoQ8. In at least some embodiments, the preparation may be administered simultaneously with or together with compositions that promote the growth or metabolism of ammonia-oxidizing microorganisms, promote the production of products or by-products of ammonia-oxidizing microorganisms, promote urease activity, or have a synergistic effect with ammonia-oxidizing microorganisms, such as ammonia, ammonium salts, urea, and urease.

[0180] The preparation may be administered with a microbiome cleansing preparation, such as a topical or systemic antibiotic. The preparation may be administered after the administration of a cleansing preparation or a bowel cleansing preparation. The preparation may be administered before or after a surgical procedure, a diagnostic procedure, or a natural event, such as childbirth. The preparation may be administered before, during, or after the placement of an implantable or invasive device.

[0181] According to one or more embodiments, the preparation can be administered as an analgesic or prophylactic agent. The preparation may be self-administered. Administration of the preparation may be done using a device.

[0182] In some embodiments, ammonia-oxidizing microorganisms, for example, a preparation of ammonia-oxidizing microorganisms, are used in amounts of approximately 10 per application, per day, per week, or per month. 3 ~10 4 CFU, 10 4 ~10 5 CFU, 10 5 ~10 6 CFU, 10 6 ~10 7 CFU, 10 7 ~10 8 CFU, 10 8 ~10 9 CFU, 10 9 ~10 10 CFU, 10 10 ~10 11 CFU, 10 11 ~10 12 CFU, 10 12 ~10 13 CFU, or 10 13 ~10 14 They are administered in doses of CFU or higher. In some embodiments, ammonia-oxidizing microorganisms are administered at a dose of approximately 10 per application or per day. 9 ~10 10 CFU, for example, approximately 1 × 10⁻⁶ 9 ~5×10 9 , 1 x 10 9 ~3×10 9 , or 1 × 10 9 ~10×10 9 It is administered in CFU doses.

[0183] In some embodiments, the ammonia-oxidizing microorganisms are administered in volumes of approximately 1–2, 2–5, 5–10, 10–15, 12–18, 15–20, 20–25, or 25–50 ml per dose. In some embodiments, the solution is approximately 10 8 ~10 9 , 10 9 ~1010 , or 10 10 ~10 11 The concentration is CFU / ml. In some embodiments, ammonia-oxidizing microorganisms are administered at a dose of 10 per day. 9 A dose of 15 ml, corresponding to the CFU / ml concentration, is administered twice.

[0184] In some embodiments, the ammonia-oxidizing microorganisms are administered once, twice, three, or four times per day. In some embodiments, the ammonia-oxidizing microorganisms are administered once, two, three, four, five, or six times per week. In some embodiments, the ammonia-oxidizing microorganisms are administered immediately after bathing. In some embodiments, the ammonia-oxidizing microorganisms are administered immediately before going to bed.

[0185] In some embodiments, ammonia-oxidizing microorganisms are administered over periods of approximately 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, and 84-91 days, for example, over approximately one month, over approximately two months, and over approximately three months. In some embodiments, ammonia-oxidizing microorganisms are administered over indefinite periods, for example, over one year, over five years, over ten years, over fifteen years, over thirty years, over fifty years, and over seventy years.

[0186] Administration of ammonia-oxidizing microorganisms to the digestive system The formulations (e.g., preparations or compositions) described herein may include those suitable for enteral delivery, such as oral administration, sublingual administration, and rectal administration. Ammonia-oxidizing microbial preparations may be administered to the gastrointestinal system for cosmetic or therapeutic purposes. For example, compositions may include those formulated for cosmetic or therapeutic use.

[0187] Enteral preparations (e.g., preparations or compositions) can be conveniently presented as unit dosage forms and can be prepared by any method known in the art of pharmaceuticals or cosmetics. Typically, the method involves attaching an active ingredient (e.g., ammonia-oxidizing microorganisms) to a pharmaceutical carrier constituting one or more auxiliary components. Usually, pharmaceutical or cosmetic preparations are prepared by uniformly and closely attaching the active ingredient to a liquid carrier, a micronized solid carrier, or both, and then, if necessary, shaping the product into the desired formulation.

[0188] Enteral preparations can be presented as individual units, each containing a predetermined amount of the active ingredient as a solution or suspension in an aqueous or non-aqueous liquid, as a powder or granules, or as an oil-in-water or water-in-oil liquid emulsion. Various pharmaceutically acceptable carriers and their formulations are described in standard pharmaceutical textbooks, for example, Remington's Pharmaceutical Sciences by E.W. Martin. Also, see Wang, Y.J. and Hanson, MA, Journal of Parenteral Science and Technology, Technical Report Vol. 10, Vol. 42: Supplement 2S, 1988; and Aulton, M. and Taylor, K., Aulton's Pharmaceutics: The Design and Manufacture of Medicines, 5th edition, 2017; Antoine, A., Gupta MR, and Stagner, WC, Integrated Pharmaceutics: Applied Preformulation, Product Design, and Regulatory Science, 2013; Dodou K. Exploring the Unconventional Routes - See also Rectal and Vaginal Dosage Formulations, The Pharmaceutical Journal, August 29, 2012.

[0189] The compositions disclosed herein can be prepared as enteral formulations. Ammonia-oxidizing microorganisms can be administered enterally for cosmetic or therapeutic purposes. Enteral formulations are generally intended to be absorbed through various epithelial and mucous membranes of the digestive system. For example, compositions can be prepared as tablets, capsules, solutions, suspensions, emulsions, or suppositories. Each dosage form can be formulated to contain one or more carriers or additives, as further detailed below. Solid dispersion forms may include tablets and capsules. Tablets can be formulated to disintegrate into granules and powders upon entering the gastrointestinal tract. Capsules can be formulated as soft-shell or hard-shell capsules. Liquid dispersion forms, such as oral dispersion forms, may include solutions, suspensions, or emulsions of the activator in a vehicle.

[0190] Compositions prepared for rectal administration can be formulated in the same way as, or differently from, tablets, capsules, solutions, suspensions, and emulsions for oral administration. However, rectal compositions may also be formulated for administration as rectal solutions, suppositories, ointments, gels, emulsions, or films. Typically, rectal solutions may be aqueous solutions, such as aqueous dispersions of activators. Rectal ointments may include an anhydrous dispersion of activators, for example, in a mineral oil-white petrolatum substrate. Rectal gels may contain polymers, such as poloxamer, xanthan gum, gellan gum, locust bean gum, and carrageenan. Rectal ointments and gels may provide longer residence times than, for example, aqueous solutions. Longer residence times may further allow for reduced dosing intervals. Rectal emulsions may contain microspheres, microcapsules, nanoparticles, nanocapsules, micelles, liposomes, niosomes, dendrimers, or cyclodextrin complexes. Rectal films may include, for example, water-soluble polymer films or polyvinyl alcohol polymer films that dissolve and release an activator upon contact with bodily fluids.

[0191] The ammonia oxidation compositions disclosed herein may contain an effective amount of AOM for purposes such as increasing the viscosity of mucus in at least a portion of the digestive system, adhering to the tissues of the digestive system, treating a digestive disorder or symptoms of a digestive disorder, improving digestion in a subject, or promoting endothelial function within the digestive system. The compositions may be administered to a substantially empty stomach or after bowel cleansing or antibiotic treatment. In some embodiments, water is administered to the subject after administration of the ammonia oxidation microbial composition. In some embodiments, several hours are allowed between administration and food consumption. Such compositions may be formulated for topical, oral, sublingual, sublabial, intraoral, rectal, or device-assisted application. Topical formulations may include, for example, enemas, bidets, cleansing solutions, sprays, aerosols, and mists. Oral formulations are typically prepared specifically for oral ingestion. Sublingual and sublabial formulations, such as tablets, strips, drops, sprays, aerosols, mists, lozenges, and effervescent tablets, can be administered orally to diffuse through the connective tissue beneath the tongue or lip. Specifically, formulations for sublingual administration may be placed under the tongue, and formulations for sublabial administration may be placed between the lip and the gums. Sublabial administration may be advantageous when the dosage form contains materials that may be corrosive to sensitive tissues beneath the tongue. Intraoral formulations can usually be held locally or applied to an oral region to diffuse through the oral mucosal tissue covering one side of the cheek. Sublingual, sublabial, and intraoral administrations can offer a faster onset of action compared to oral administration because the active ingredient enters the bloodstream directly, avoiding first-pass metabolism. Rectal administration can be performed by inserting the formulation into the rectal lumen, with or without the use of a device. Device-assisted applications may include, for example, delivery via an applicator or insertable applicator, catheter, or replenishment tube, or delivery with an endoscope or ultrasound. Appropriate applicators include liquid formulation valves and dispensers, as well as applicators into which solid formulations can be inserted.

[0192] The time of action of the formulations disclosed herein may depend on the formulation and may range from seconds to minutes to hours. For example, tablets and rectal solid dosage forms may take effect within minutes. Oral tablets may take effect within minutes. Suppositories, solutions, and suspensions may take effect within minutes or hours. Powders, granules, tablets, and capsules may take effect within minutes to hours. Regulated-release tablets may take effect within minutes to hours. Enteric-coated formulations may take effect within hours. The release time of the formulations disclosed herein may depend on the formulation and may range from minutes to hours to days. For example, a dosage form may be formulated to provide immediate release within minutes or extended release within hours. Certain dosage forms may provide extended release within days or months.

[0193] The ammonia-oxidizing microbial compositions disclosed herein may be in the form of tablets or capsules. Tablets may typically contain disintegrants to facilitate the disintegration of the tablet into smaller particles, such as granules and powder particles, and to facilitate dissolution and absorption. Tablets may be coated to provide a protective barrier against environmental factors for drug stability, to mask an unpleasant drug taste, or to protect the drug from acidic conditions in the stomach. Tablets may further contain fillers, binders, bulking agents, and diluents to increase the size of the tablet, binders to promote aggregation, anti-adhesives and anti-sticking agents to reduce film formation, adhesion, and sticking, flow promoters and flow aids to reduce interfacial aggregation and friction, wetting agents, solubilizers, stabilizers, colorants, sweeteners, and flavorings. Capsules may typically contain fillers, such as liquid, gel-like, semi-liquid, semi-solid, or microemulsion formulations, and coatings, such as hard-shell or soft-shell coatings, which may include gelling agents, hypermellose, or polymeric materials. Hard-shell capsules can be filled with solids (e.g., powders, granules, pellets, tablets, and capsules), semi-solids (e.g., gels, pastes, and thermosetting polymers), and liquids (e.g., solutions, suspensions, emulsions, and microemulsions). Soft-shell capsules can be filled with liquids and semi-liquids. Capsules may also contain plasticizers to reduce brittleness (e.g., glycerin, sorbitol, propylene glycol, poly(ethylene glycol), hydrogenated sugars, polysaccharides, and sorbitan), processing aids to improve gelation and precipitation (e.g., carrageenan and cations), buffers, surfactants, wetting agents, lubricants, colorants, opacifiers, and fragrances. The coating may readily rupture or dissolve after administration. The tablets and capsules may further contain, for example, a polymer-based component or coating film, which is rapidly soluble for immediate delivery, controlled release, delayed release, or sustained release.

[0194] The compositions disclosed herein can be specifically formulated to be enteric, for example, to be resistant to the harsh and acidic environment of the stomach. In some embodiments, the ammonia-oxidizing microbial composition is placed in a gel or agar matrix formulated to withstand an acidic environment (e.g., the acidic environment of the stomach) and disintegrate at the physiological pH of the intestine. The formulation can be coated with a composition that is formulated to withstand a particular gastrointestinal environment and deliver an active agent, for example, by disintegrating in the physiological environment associated with specific gastrointestinal tissues.

[0195] The ammonia-oxidizing microbial composition can be in a liquid or semi-liquid dispersed form, for example, a syrup or a linctus. The liquid and semi-liquid dispersions can include a vehicle for delivery, a density modifier to increase the density of the dispersion medium, for example, to match the density of the dispersed particles, and a viscosity modifier to increase the viscosity and reduce sedimentation. The dispersion can include a wetting agent, a surfactant to reduce the interfacial energy between the vehicle and the dispersed particles, a flocculating agent to control the level of loose aggregation or floc formulation, a humectant to reduce the rate of moisture loss of the product, as well as buffering agents, sweeteners, flavors, and colorants disclosed herein.

[0196] Typical compositions may include, for example, microcrystalline cellulose for bulking, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, dicalcium phosphate, starch, magnesium stearate and / or lactose and / or other additives, binders, extenders, disintegrants, diluents and lubricants, mannitol, lactose, sucrose, and / or cyclodextrin, including suspensions. Such formulations may also include high molecular weight additives such as cellulose (Avicel) or polyethylene glycol (PEG). Such formulations may also include additives to assist in mucoadhesion, such as agents for controlling release, such as hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), sodium carboxymethylcellulose (SCMC), maleic anhydride copolymers (e.g., Gantrez), and polyacrylic copolymers (e.g., Carbopol 934). Lubricants, flow promoters, flavors, colorants, and stabilizers can also be added to facilitate manufacture and use. The surfactant can be an amphoteric surfactant, a nonionic surfactant, or an anionic surfactant.

[0197] Suppositories include solid dosage forms intended for introduction into the rectal cavity. Suppositories can melt when introduced into the rectal cavity and release the active agent. The delivery rate of the active agent can be affected by the choice of pharmaceutically acceptable carrier or suppository base. Suitable suppository bases include fatty bases and aqueous bases. Suitable fatty base formulations can include cocoa butter, whale oil, triglycerides derived from synthetic triglycerides or hydrogenated vegetable oils, palm oil, palm kernel oil, or coconut oil. Suitable aqueous base formulations can include glycerinated gelatin or polyethylene glycol polymers. Suppository formulations may further include absorption enhancers.

[0198] Solid rectal dosage forms containing the compositions disclosed herein can be formulated as wet tablets, compressed tablets, or capsules. Wet tablets can be made in various shapes, for example, conical, and can be prepared in a manner similar to that of suppositories. Compressed tablets can be made in various shapes and can be prepared by compression. Compressed rectal tablets may typically contain formulations and additives similar to those of oral tablets. Accordingly, compressed rectal tablets disclosed herein can be formulated to contain one or more of the following: fillers, binders, expanders, diluents, disintegrants, lubricants, anti-adhesive and anti-sticking agents, flow enhancers and flow agents, wetting agents, solubilizers, drug release modifiers, stabilizers, and colorants. Rectal capsules can be prepared in a manner similar to that of gelatin oral capsules. For example, the rectal capsules disclosed herein may be formulated to contain one or more solid, semi-solid, and liquid fillers, plasticizers, processing aids, surfactants, colorants, opacifiers, and preservatives. The body of the rectal capsule may contain gelatin, hypermellose, hydroxypropyl methylcellulose, hydroxypropyl starch, starch products, and pullulan.

[0199] Ointments, foams, and gels can typically be formulated to be more viscous than aqueous solutions and to have a longer residence time in body cavities, such as the rectal cavity. Such viscous liquid formulations may contain a gel or gelling agent. For example, the gelling agent may be a thermoplastic gel. A thermoplastic gel may be liquid at low or room temperature and may transform into a gel when inserted into a body cavity, such as the rectum or colon. The gel or gelling agent may facilitate the administration and positioning of the dosage form. For example, the gel or gelling agent may prevent the dosage form from leaking out of the body cavity. Thermoplastic polymers include poloxamer. Mucosal adhesive polymers include sodium alginate. The gel or gelling agent may further contain a solubility enhancer, such as hydroxypropyl beta-cyclodextrin. Gel formulations may include, but are not limited to, agar, silica, polyacrylic acid (e.g., Carbopol®), carboxymethylcellulose, starch, guar gum, alginate, or chitosan.

[0200] Solutions containing the compositions disclosed herein can be formulated, for example, as enemas or bidets for rectal delivery. Typically, enemas or bidets may be aqueous solutions containing the active agent. Enemas can be administered to reach deep parts of the rectal lumen, such as the colon, or superficial parts of the rectal lumen, such as the rectum. In some embodiments, enemas are administered in volumes of 2 L or less. (US Department of Health) Human Services is hesitant to use douches, citing several risks, including irritation, bacterial infection, and inflammatory diseases of the genitourinary tract. However, in some specific circumstances, physicians may still prescribe douches for medical reasons. Since disinfectants used during douches can disrupt the natural balance of microorganisms in body cavities and cause infection, such physician-prescribed douches may be administered in combination with the ammonia-oxidizing microbial compositions disclosed herein. Furthermore, the ammonia-oxidizing microbial compositions disclosed herein may be administered in a douche as a first or second therapy, for example, in combination with one or more additional treatments. In addition, the solutions may be formulated as sprays, aerosols, or mists for local delivery to the rectal area.

[0201] A typical composition may include one or more additives, such as absorption enhancers and penetration enhancers, analgesics, topical analgesics, antifungal agents, anti-inflammatory agents, steroids and corticosteroids, thermoplastic gels, preservatives, antioxidants, buffers, chelating agents, ion exchangers, solubilizers, suspending agents, thickeners, surfactants, wetting agents, isotonic agents, and vehicles for appropriate drug delivery. Absorption enhancers and penetration enhancers can improve the ability of activators to be absorbed by many different mechanisms. Analgesics and topical analgesics can be used to relieve pain and / or reduce discomfort in the target. Steroids and corticosteroids can help reduce inflammation. Thermoplastic gels can improve the positioning and retention time of activators. Antioxidants can reduce the oxidative degradation of activators. Buffers can maintain the desired pH of the composition and / or enhance the solubility or stability of the composition. Chelating agents may include complex group metals that catalyze oxidation reactions in the composition. Ion exchangers can control the release of activators by ion exchange mechanisms. Solubilizers can increase the solubility of activators or other additives. Suspensioners and thickeners can increase the viscosity or density of the composition, thereby increasing the retention and residence time of activators in the digestive system, e.g., the oral cavity or rectal lumen. Surfactants, including cationic, anionic, and nonionic surfactants, as well as wetting agents, can act to wet insoluble hydrophobic activators or other additives. Isotonic agents can provide an isotonic solution with urogenital fluids. Vehicles, e.g., water-based or fatty-based materials, can provide size for appropriate activator delivery.

[0202] Possible additives include proteins such as human serum albumin or plasma preparations. If desired, the pharmaceutical composition may also contain small amounts of non-toxic auxiliary substances such as humectants or emulsifiers, preservatives, and pH buffers, e.g., sodium acetate or sorbitan monolaurate. In some embodiments, additives, such as pharmaceutically acceptable or cosmetically acceptable additives, may include anti-adhesives, binders, coatings, disintegrants, fillers, fragrances, colorants, lubricants, flow enhancers, adsorbents, preservatives, or sweeteners. In some embodiments, the preparation may be substantially free of additives. In some embodiments, the preparation may be substantially free of one or more of the compounds or substances listed in this disclosure.

[0203] Ammonia-oxidizing microbial compositions can be administered in forms suitable for immediate or delayed release, for example. Suitable examples of immediate-release formulations include topical formulations, oral formulations, intraoral formulations, sublingual formulations, sublabial formulations, and rectal delivery formulations. Topical formulations for immediate release may include, for example, solutions, suspensions, emulsions, foams, gels, and ointments. Topical formulations can be formulated for immediate release to avoid complications from clearance by body fluids, for example, in the rectal lumen. Rectal delivery formulations for immediate release include, for example, suppositories, solid forms for the rectum, and films. Each of the rectal delivery formulations can be formulated to change phase and release the activator upon contact with body fluids in the body lumen. For example, suppositories and tablets can be formulated for immediate release to avoid challenges arising from the expulsion of the dosage form and weak adhesion to the rectal lumen membrane. Certain target membranes, such as the rectal mucosa, allow for rapid absorption of the activator, while a high concentration of localized vascular structures can facilitate systemic uptake (update). Orally delivered formulations for immediate release include liquid dispersions, tablets, and capsules. Specifically, oral formulations, sublabial formulations, and sublingual formulations can result in the immediate release of the active ingredient into the systemic circulation, for example, by facilitating uptake through the oral mucosa, sublingual tissue, and sublabial tissue, which can bypass first-pass metabolism.

[0204] Controlled-release oral formulations can be prepared as liquid or solid dispersions. Suitable examples of sustained-release systems include suitable gelling or polymer materials (polymer-based nuclei or coating films), e.g., semipermeable polymer matrices in the form of molded articles, e.g., films or microcapsules; suitable hydrophobic materials, e.g., as emulsions in acceptable oils; or ion exchange resins. Pharmaceutical compositions may be in the form of particles containing one or more of biodegradable polymers, polysaccharide gelling polymers and / or bioadhesive polymers, or amphiphilic polymers. These compositions exhibit certain biocompatibility features that enable controlled release of the active ingredient.

[0205] In some embodiments, controlled-release rectal formulations can be formulated as ointments, gels, foams, or emulsions. Pharmaceutically extended-release compositions, such as rectal delivery compositions, can be formulated with one or more mucosal adhesives, such as a mucosal adhesive gel or a dry mucosal adhesive tablet. The mucosal adhesives can aid adhesion to the rectal lumen, e.g., the rectal mucosa. Similarly, solid dosage forms, such as suppositories, tablets, capsules, or films, can be formulated with one or more mucosal adhesives that can enhance the positioning of the dosage form within the rectal lumen or remain present when a portion of the solid dosage form dissolves or disintegrates. For example, a solid dosage form can be formulated to dissolve rapidly upon contact with body fluids and transform into a mucosal adhesive viscous solution that adheres to the oral mucosa or rectal mucosa and is gradually washed away without requiring removal. Sublabial dosage forms can result in controlled or extended release of the active ingredient. For example, sublabial administration between the upper lip and gums can prevent the preparation from being swallowed with saliva, allowing for delayed, long-term release of the drug directly into the connective tissue and associated vascular structures.

[0206] In some non-limiting embodiments, the preparation may be one or more of the following: substantially odorless, colorless, substantially free of side effects, non-toxic, well-tolerated, pose no adverse effects when released into the environment, pose no risk of promoting antibiotic resistance, and have a physiology that allows for positive interactions with various human gastrointestinal microbiomes under normal and diseased conditions.

[0207] Ammonia-oxidizing microbial compositions can be administered in a form suitable for, for example, a topical or systemic treatment. Topical effects may be achieved, for example, by rapid absorption through the tissues of the gastrointestinal system, and systemic effects may be achieved, for example, by drug absorption through various epithelial and mucous membranes of the gastrointestinal tissues. The compositions disclosed herein can be administered to treat topical inflammatory diseases, symptoms of topical or systemic inflammatory diseases, or side effects resulting from topical or systemic inflammatory diseases. Suitable examples of local gastrointestinal conditions or disorders that can be treated with the compositions disclosed herein include irritable / inflammatory bowel syndrome, Crohn's disease, colitis, nectrotizing panentotitis, ulcers, ulcerative colitis, celiac disease, gluten sensitivity, lactose intolerance, food and / or beverage intolerance, heartburn, acid reflux, symmetric motility disorder, small intestinal bacterial overgrowth (SIBO), pancreatitis, appendicitis, gastritis, malabsorption disorders, and gastroenteritis. Leaky gut syndrome may also be addressed. The administration can be used to promote GI health. The gastrointestinal condition of the subject may be affected, for example, by altering, modifying, or modulating the composition of the GI microbiome, for example, by changing the ratio of microorganisms therein, for example, in the gastrointestinal tract or GI tract. The administration can be used before travel to reduce the possibility of GI impairment when exposed to unfamiliar foods, beverages, and / or microorganisms. In some embodiments, ammonia-oxidizing microbial compositions can be administered in a form suitable for treating certain infectious and inflammatory disorders, such as bacterial infections, fungal infections, viral infections, itching, local inflammation, and wound healing. For example, ammonia-oxidizing microbial compositions can be administered to treat inflammation associated with surgical or diagnostic procedures, dental procedures, catheter-based transfers (e.g., transfer of material inside, outside, or between two locations in the body), enteral nutrition (e.g., feeding tubes), stent-related inflammation, or inflammation associated with any foreign body introduced into the gastrointestinal or biliary system.Ammonia-oxidizing microorganisms can be administered to treat localized symptoms of gastrointestinal conditions, disorders, or systemic disorders, such as stomach pain, stomach cramps, gas, constipation, discomfort, changes in bowel habits, and diarrhea, or side effects associated with such conditions, disorders, or systemic disorders, such as hyperammonemia, as well as biliary disorders, such as hepatic failure and acute hepatic failure. In at least some embodiments, administration of ammonia-oxidizing microorganism compositions may reduce symptoms or side effects associated with gastrointestinal conditions or disorders, such as bloating, diarrhea, gas, stomach pain, stomach cramps, or borborygmus.

[0208] Examples of systemic conditions that can be treated with the compositions disclosed herein include headaches, cardiovascular diseases, connective tissue disorders, inflammation, immune responses and autoimmune disorders, liver diseases, infections, neurological disorders, psychiatric disorders, nitric oxide disorders, urea cycle disorders, congestion, vasodilation disorders, skin diseases, wound healing, intestinal disorders, reactions to insect bites, eye disorders, and certain viral, bacterial, and fungal infections. For example, systemic conditions that can be treated with the compositions disclosed herein include cardiovascular diseases such as cardioprotection, heart failure, hypertension, and pulmonary arterial hypertension; immune responses and autoimmune disorders such as alopecia and vitiligo; liver diseases such as non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH); neurological and psychiatric disorders such as depression, insomnia, and diabetic neuropathy; nitric oxide disorders such as erectile dysfunction; and, for example, pressure ulcers and care in nursing facilities. This includes wound healing from burns, diabetic ulcers, such as foot ulcers, venous leg ulcers, fungal infections, and mouth pain; skin diseases and disorders such as hyperhydrosis, pruritus, corns, and subtypes of corns; eye disorders such as blepharitis, dry eye, macular degeneration, and glaucoma; intestinal disorders such as gluten sensitivity, irritable / inflammatory bowel disease, Crohn's disease, colitis, and necrotizing enterocolitis; and vasodilatory disorders such as Raynaud's disease, thermoregulation, and migraines. Certain viral, bacterial, and fungal infections, including infections caused by human papillomavirus (HPV), yeast infections, tinea versicolor, onychomycosis, tinea pedis / fungal infections, tinea cruris, tinea cruris, onychomycosis, dandruff, athlete's foot, sinusitis, otitis media, methicillin-resistant Staphylococcus aureus (MRSA), staphylococcal infections, and bacterial vaginosis, can be treated with the formulations disclosed herein.Further systemic conditions that can be treated with the compositions disclosed herein include systemic inflammations such as eczema, e.g., eczema in adults and children, urticaria, idiopathic urticaria, lichen planus, insect bites, e.g., allergic reactions to mosquitoes and Demodex folliculorum mites, poison ivy reactions, itching, keratosis pilaris, pharyngitis, pemphigus, psoriasis, rosacea, folliculitis and subtypes of folliculitis, hidradenitis suppurativa (supportiva), perioral dermatitis, lupus rash, seborrheic dermatitis, e.g., seborrheic dermatitis in adults and infants, acne, e.g., acne in adolescents, acne in adults, and cystic acne, diaper rash, occupational hand dermatitis, sunburn, and dermatomyositis. Furthermore, the compositions disclosed herein may be delivered or applied to treat certain cosmetic indications, including, but are not limited to, contact dermatitis, diaper odor, adult and child body odor, feminine odor, dryness, hardened nails, body odor, oily skin, razor burn, skin appearance, numerous blemishes, skin hydration, and sunspots. The compositions disclosed herein may be applied as insecticides or antimicrobial agents.

[0209] The compositions disclosed herein can be further formulated as combination therapies. For example, tablets or capsules may contain different parts formulated for combination therapy. The first and subsequent therapeutic treatments may be provided in single doses, prepared in individual dosage forms, administered simultaneously, or administered individually. Individual dosage forms may be administered via the same mode of administration, for example, through the gastrointestinal system, or via alternative modes of administration, for example, orally, intranasally, topically, ocularly, through the auditory system, through the genitourinary system, through the respiratory system, or by injection. For example, a combination therapy may contain ammonia-oxidizing microorganisms to treat an inflammatory disease or condition. Individual dosage forms may be administered by surgical or diagnostic procedures. Individual dosage forms may be administered in combination with surgical or diagnostic procedures. In some embodiments, for example, an ammonia-oxidizing microorganism composition prepared for enteral administration may be formulated for combination therapy with anti-inflammatory properties. Typically, the compositions disclosed herein can be formulated for combination therapy with drugs or compounds approved for or commonly used to treat diseases, disorders, conditions, their symptoms, or side effects, such as gastrointestinal diseases, disorders, conditions, their symptoms, or side effects. In at least some embodiments, the preparations are formulated for administration in combination with fecal microbiota transplantation (FMT). The preparations can be formulated for administration in combination with colon cleansing or antibiotics to "cultivate" and enable the colonization of ammonia-oxidizing microorganisms.

[0210] Preparations for administration to the gastrointestinal system can be formulated for targeted delivery to specific depositional or target tissues. In some embodiments, a preparation can be administered to a first tissue such that the preparation or a product of the preparation, e.g., ammonia-oxidizing microorganisms or nitric oxide, is transported to a second tissue. The first tissue may be depositional tissue. The second tissue may be target tissue. The depositional and target tissues may be the same or different tissues. In some embodiments, the depositional tissue, target tissue, or both may be tissues of the gastrointestinal system. The preparation or a product of the preparation can be delivered locally or systemically, for example, to the depositional or target tissue, or intracirculating.

[0211] Preparations for intra-gastrointestinal administration can be formulated for targeted delivery to specific gastrointestinal tissues. Gastrointestinal deposition or target tissues include the mouth, stomach, salivary glands, oral cavity, pharynx, tongue, esophagus, liver, gallbladder, common bile duct, colon (transverse colon, ascending colon, and / or descending colon), cecum, appendix, rectum, anus, pancreas, pancreatic duct, large intestine, small intestine (duodenum, jejunum, and / or ileum), parotid gland, submandibular tissue, and sublingual tissue. In some embodiments, systemic uptake in the stomach may be difficult due to the acidic environment of the stomach. Systemic uptake in the liver may be difficult due to the metabolism of the compound upon entering the liver. Compositions can be specifically formulated to bypass the stomach and / or liver and deliver the activator to another target tissue. For example, a composition can be formulated to bypass hepatic metabolism and deliver the target active ingredient into the oral cavity or lower rectum, or it can be specifically formulated to withstand the acidic environment of the target and deliver the activator to the small or large intestine. Typically, the gastrointestinal route can produce systemic effects through the large surface area and blood supply available to the gastrointestinal tissues.

[0212] The rectal route may be useful in children and the elderly, as well as in patients who cannot take medication orally due to nausea, vomiting, or unconsciousness. Rectal administration can be used for the delivery of drugs for the prevention of infection and treatment of inflammation before and after surgery. Preparations for rectal administration can be formulated for targeted delivery to specific rectal tissues. Rectal depositional or target tissues may include surface tissues, such as the buttocks, anus, and perianal area, as well as internal tissues, such as the rectum, colon, large intestine, small intestine, and anal sphincter. Preparations for rectal administration can further be formulated for targeted delivery to surrounding tissues, such as the pelvic floor muscles and prostate. Targeted delivery to desired rectal tissues may be important in providing appropriate treatment. Rectal dosage forms for targeted delivery to the upper rectum and colon can be absorbed, for example, by the surface rectal vein and delivered to the liver. Therefore, rectal dosage forms for targeted delivery to the upper rectum and colon can be formulated for hepatic metabolism. Rectal formulations for targeted delivery to the lower rectum are absorbed via local lower veins and can bypass the liver. Formulations unsuitable for hepatic metabolism can be prepared for targeted delivery to the lower rectum.

[0213] It is surprising that, according to one or more embodiments, AOM can grow and reproduce in the environment of the digestive system in question. The stomach, for example, may generally be characterized by being inherently anaerobic. In some embodiments, the mucus of the stomach wall may shift to higher pH levels. In some embodiments, surprisingly, AOM may act as a key species in regulating the viscosity of mucus in the digestive system. While we do not wish to be bound by any particular theory, nitrite levels may play a role in regulating the viscosity of mucus in the GI system. Mucus may play an important barrier role, for example, in preventing infection within the GI system. In some non-limiting embodiments, the mucus layer may include two regions: a highly adhesive, sterile inner layer that may indirectly contact the digestive tract wall, and a less rigid layer made of low-density gel-forming peptides. In some non-limiting embodiments, mucus release may be increased, which may facilitate the digestion of disulfide-containing substrates, for example, gluten. Disulfide isomerase, a protein, may also be released. In some embodiments, a more viscous mucus may better seal the GI system wall, such as the gastrointestinal wall, and reduce inflammation caused by facultative anaerobic bacteria. In some non-limiting embodiments, Hilicobacter pylori may be reduced by AOM colonization.

[0214] In at least some embodiments, the effects of superoxide can be weakened. In some embodiments, AOM can suppress undesirable aerobic and facultative anaerobic bacteria. In some embodiments, the viscosity of mucus can be increased while suppressing facultative anaerobic bacteria by depriving them of oxygen and nitrates. In at least some embodiments, AOM can survive in mucus adhering to the walls of the GI system. According to one or more embodiments, AOM can surprisingly proliferate in the transient environment of the GI system despite long doubling times. In some embodiments, AOM surprisingly does not promote diarrhea and rapid, uncontrollable GI system excretion, despite producing NO and nitrites that can relax the smooth muscles of the GI system and have this effect. In some embodiments, AOM can reduce constipation, for example, by improving the compatibility of peristalsis and nutrient absorption. In some embodiments, combination therapy may involve heterotrophic probiotics such as Lactobacilli. According to one or more embodiments, AOM can generate physiologically beneficial and / or significant levels of NO, nitrite, and / or NO precursors from ammonia and oxygen within the GI system. While we do not wish to be bound by any particular theory, the GI system in question may feature a variety of oxygen and / or NO gradients. In at least some embodiments, integrating AOM into the GI system can delay or halt the progression of various local and systemic GI conditions, after which AOM further leads to an optimized state of health. In some embodiments, recovery of AOM after alcohol consumption can reduce and / or prevent certain adverse effects in the visceral system and the liver, among others.

[0215] According to one or more embodiments, blood flow from the stomach passes through the liver. While we do not wish to be bound by a specific theory, nitric oxide may be a normal regulator of blood flow, mitochondrial biosynthesis, ATP status, and the allocation of resources to immediate consumption or repair. Inflammatory states in the GI system may be associated with adverse effects on the liver, e.g., high portal pressure, primary sclerosing cholangitis, non-alcoholic steatohepatitis, non-infectious hepatitis, as well as multiple systemic inflammatory states such as asthma, multiple sclerosis, chronic kidney disease, psoriasis, pericarditis, and arthritis. Similarly, diabetes, stroke, and heart disease may be associated with chronic inflammatory disorders of the gastrointestinal tract. Increased blood volume and / or hypercirculation in visceral systems may result. Decreased renal blood flow, which can lead to renal failure, may occur. Prevention of inflammation in the GI system by the establishment of AOM according to various embodiments may also reduce the incidence of liver damage associated with GI inflammation and other systemic inflammatory states. In at least some embodiments, AOM preparations can be administered to premature infants and / or neonates in the GI system. For example, necrotizing enterocolitis can be treated or prevented in these subjects.

[0216] According to one or more embodiments, AOM can be incorporated into a variety of food and / or beverage products that are generally suitable for it in terms of, for example, temperature, salt, alcohol, or preservative levels. In some embodiments, a culture of AOM can be applied to food or beverages before consumption, for example, by spraying. Some non-limiting examples of such food products include yogurt, milk, cheese, salads, fruits, and bread. With respect to liquids containing AOM, at consumption, water may be absorbed through the stomach wall, AOM may be filtered out, and may be trapped in the mucus layer. Ammonia release hotspots may be converted by AOM into nitrite hotspots. Acidified nitrites may act as antimicrobiome agents, for example, to prevent stomach infections. As stomach contents descend through the GI system, the mucus layer also descends, and therefore the mucus layer containing AOM may advance downward through the GI system for a variety of favorable results consistent with this disclosure.

[0217] Use of a product compatible with the microbiota, with administration of ammonia-oxidizing microorganisms Products compatible with the microbiota can be used with the preparations and methods disclosed herein. Various products can be considered "microbiota-safe" or "microbiota-compatible." Examples of microbiota-safe products are disclosed in International (PCT) Patent Application Publication No. WO2017 / 004534 (International (PCT) Patent Application No. PCT / US / 2016 / 040723, filed July 1, 2016), which is hereby incorporated by reference in its entirety for all purposes. Some products that are microbiota-safe can be, in essence, cosmetic or therapeutic. According to one or more embodiments, a microbiota-safe product can be used in combination with microorganisms, such as non-pathogenic microorganisms, such as ammonia-oxidizing microorganisms, and can then be used in the form of a preparation or composition for application to a subject. The ammonia-oxidizing compositions disclosed herein can be administered for cosmetic or therapeutic indications, together with a microbiota-safe or microbiota-compatible product.

[0218] According to one or more embodiments, a preparation, composition, formulation, or product containing ammonia-oxidizing microorganisms, for example for cosmetic use or therapeutic use, can be considered to be, in itself, microbiota-safe. In other embodiments, a preparation containing ammonia-oxidizing microorganisms can be used with a microbiota-safe product. In some embodiments, a preparation containing ammonia-oxidizing microorganisms can be mixed with a microbiota-safe product or, alternatively, administered simultaneously. In other embodiments, a preparation containing ammonia-oxidizing microorganisms can be in a different or separate form from a microbiota-safe product, but can also be combined with a microbiota-safe product. In some embodiments, a microbiota-safe product is used alone. Preparations of ammonia-oxidizing microbial compositions for use with a microbiota-safe product can be formulated for cosmetic use or therapeutic use.

[0219] Products that are harmless to or biomedically compatible with the biome can be used in conjunction with ammonia-oxidizing microbial preparations formulated for any mode of delivery, for example, for targeted delivery to a subject, for example, a target tissue, target region, target system, or target organ of the subject. For example, an ammonia-oxidizing microbial preparation for use with a biome-harmless product can be formulated for delivery to the eyes, ears, nose, genitourinary system, respiratory system, or digestive system of the subject. In some embodiments, an ammonia-oxidizing microbial composition for use with a biome-harmless product can be formulated for targeted delivery based on the condition or disorder of the subject. For example, formulations for targeted delivery can be based on the desired local or systemic effect to be achieved, for example, a local or systemic therapeutic effect or a cosmetic effect.

[0220] Cosmetic products that may be used in conjunction with this disclosure and are biome-safe include: baby products, e.g., baby shampoo, baby lotion, baby oil, baby powder, baby cream; bath preparations, e.g., bath oil, tablets, salt, bubble bath, bath capsules; eye makeup preparations, e.g., eyebrow pencil, eyeliner, eyeshadow, eye lotion, eye makeup remover, mascara; fragrance preparations, e.g., cologne, eau de toilette, perfume, powder (dusting and talcum), sachets; hair preparations, e.g., hair conditioner, hair spray, hair straightener, permanent waver, rinse, shampoo, tonic, hair styling products, styling aids, wave setters; hair coloring preparations, e.g., hair dyes and hair colors, hair dyes, coloring hair rinses, coloring hair shampoos, coloring hair lighteners, hair bleach; makeup preparations, e.g., face powder, foundation, leg and body paint, lipstick, makeup base The products include any one or more of the following: rouge, blush, makeup fixatives; manicure preparations, e.g., base coats and undercoats, cuticle softeners, nail creams and lotions, nail extenders, nail polish and enamel, nail polish and enamel removers; oral hygiene products, e.g., toothpaste, mouthwash and breath fresheners; bath soaps, e.g., foaming body washes and detergents, deodorants, bidets, women's hygiene deodorants; shaving preparations, e.g., aftershave lotions, beard softeners, talcum, pre-shave lotions, shaving creams, shaving soaps; skincare preparations, e.g., cleansers, depilators, powders and sprays for face and neck, body and hands, and feet, moisturizers, night preparations, paste masks, and skin fresheners; and tanning preparations, e.g., gels, creams, liquids, and indoor tanning preparations, which may be included in or contained within.

[0221] Products described herein, such as microbiome-compatible cosmetic products, such as shampoos, conditioners, and cleansers, can be used in conjunction with the treatment of a condition, disease, or disorder. These cosmetic products can be used in conjunction with the administration of ammonia-oxidizing microorganisms for therapeutic or cosmetic purposes. For example, microbiome-compatible cosmetic products can be used throughout a therapeutic or cosmetic period in which ammonia-oxidizing bacteria are administered to a subject. Microbiome-compatible cosmetic products can be used for a period of time before initiating treatment of a therapeutic or cosmetic condition via the administration of ammonia-oxidizing bacteria to a subject. Microbiome-compatible cosmetic products can be used for a period of time after initiating treatment of a therapeutic or cosmetic condition via the administration of ammonia-oxidizing bacteria to a subject. Microbiome-compatible cosmetic products can be used for a period of time after discontinuation of therapeutic or cosmetic treatment of a condition via the administration of ammonia-oxidizing bacteria to a subject.

[0222] In some embodiments, the subject may wait for a certain period of time before applying one or more cosmetic products and administering ammonia-oxidizing microorganisms. In other embodiments, the subject may wait for a certain period of time before administering ammonia-oxidizing microorganisms and applying one or more cosmetic products.

[0223] The waiting period for the subject may be approximately 1 minute, 5 minutes, 10, 15, 20, 25, 30, 45, 60, 90, 120 minutes, or 3 hours, 4, 5, 6, 7, 8, 12, 18, or 24 hours after the application of one or more cosmetic products and before the administration of ammonia-oxidizing microorganisms.

[0224] The waiting period for the subject after administration of the ammonia-oxidizing microorganism and before application of one or more cosmetic products may be approximately 1 minute, 5 minutes, 10, 15, 20, 25, 30, 45, 60, 90, 120 minutes, or 3 hours, 4, 5, 6, 7, 8, 12, 18, 24 hours.

[0225] While specific embodiments of the present invention have been considered, the above specification is illustrative and not limiting. Many variations of the present invention will become apparent to those skilled in the art when considering this specification and the claims below. The entire scope of the present invention should be determined by referring to the claims together with the entire scope of equivalents, and to this specification together with such variations.

[0226] A particular embodiment falls within the scope of the following claims. The present invention provides, for example, the following items: (Item 1) A method for introducing ammonia-oxidizing microorganisms (AOM), The preparation containing AOM is administered enterally to the subject. A method that includes this. (Item 2) The method described in any of the preceding items, wherein enteral administration includes oral, intraoral, sublabial, or sublingual administration. (Item 3) A method for implementing AOM The preparation containing AOM is administered rectally to the subject. A method that includes this. (Item 4) The method described in any of the preceding items, wherein rectal administration includes administration via suppositories or enemas. (Item 5) A method according to any of the preceding items, wherein rectal administration is accompanied by a fecal microbiota transplantation procedure. (Item 6) A method for immobilizing AOM in the target digestive system, Administer an effective amount of the preparation containing AOM to the target digestive system. A method comprising the AOM being immobilized in the target tissue of the digestive system. (Item 7) A method for improving digestion in the subject, To administer an effective amount of a preparation containing AOM to the subject, thereby improving digestion in the subject. A method that includes this. (Item 8) A method for treating digestive disorders in a subject, Administer an effective amount of a preparation containing AOM to the subject to treat the gastrointestinal disorder. A method that includes this. (Item 9) The method according to any of the preceding items, wherein the amount and / or frequency of administration is sufficient to increase the viscosity of mucus in at least a portion of the digestive system of the subject. (Item 10) The method according to any of the preceding items, wherein the preparation containing the AOM is administered by ingestion into the target digestive system. (Item 11) The method according to any of the preceding items, wherein the subject has a substantially empty stomach when the preparation is administered. (Item 12) The method according to any of the preceding items, wherein the preparation is administered after the administration of an antibiotic or a bowel cleansing preparation. (Item 13) A method according to any of the preceding items, wherein the target percentage of AOM to be administered is transferred to the digestive system of the subject. (Item 14) The method according to any of the preceding items, further comprising administering water to the subject after administering the preparation. (Item 15) The method according to any of the preceding items, wherein administration of the preparation results in increased tolerance, decreased sensitivity, and / or improved nutrient uptake related to the food or beverage consumed by the subject. (Item 16) The method described in any of the preceding items, wherein the digestive disorder is an inflammatory state. (Item 17) The method according to any of the preceding items, wherein the inflammatory condition is colitis, necrotizing enterocolitis, inflammatory bowel disease, ulcer, Crohn's disease, ulcerative colitis, celiac disease, gluten sensitivity, heartburn, pancreatitis, appendicitis, gastritis, gastroenteritis, irritable bowel syndrome, or a dental or periodontal condition. (Item 18) The method according to any of the preceding items, wherein the gastrointestinal disorder is related to lactose, food or beverage intolerance, SIBO, malabsorption, biliary duct disorder, reflux, or integrative motility disorder. (Item 19) The method described above, in which the inflammatory condition is accompanied by the delivery of a substance via a catheter, such as enteral nutrition, as described in one of the preceding items. (Item 20) The method according to any of the preceding items, wherein the administration is before or after a medical procedure, such as a catheterization, endoscopy, or colonoscopy procedure, or a dental procedure. (Item 21) The method according to any of the preceding items, wherein the inflammatory condition is caused by infection with one or more of the following microorganisms: H. pylori, C. diff, cholera, amoebic dysentery, Y. enterocolitica, S. enteritidis, S. typhimurium, Shigella sonnei, and E. coli. (Item 22) The method according to any of the preceding items, wherein the digestive disorder is characterized by constipation or diarrhea. (Item 23) The method according to any of the preceding items, wherein the aforementioned gastrointestinal disorder is characterized by hyperammonemia. (Item 24) The method according to any of the preceding items, wherein the gastrointestinal disorder includes liver failure, for example, acute or chronic liver failure. (Item 25) The method according to any of the preceding items, wherein the deposited tissue, target tissue, or both are the mucous membrane of the subject. (Item 26) The method according to any of the preceding items, wherein the deposited tissue, target tissue, or both are related to the stomach of the subject. (Item 27) The method according to any of the preceding items, wherein the deposited tissue, target tissue, or both are the salivary glands, oral cavity, pharynx, tongue, esophagus, liver, gallbladder, common bile duct, colon (transverse colon, ascending colon, and / or descending colon), cecum, appendix, rectum, anus, pancreas, pancreatic duct, large intestine, or small intestine (duodenum, jejunum, and / or ileum). (Item 28) The method according to any of the preceding items relating to the target tissue for the desired local effect. (Item 29) The method according to any of the preceding items, wherein the target tissue is related to the desired systemic effect. (Item 30) The method according to any of the preceding items, wherein the desired systemic effect involves the treatment of one or more of the following conditions: headache, cardiovascular disease, inflammation, immune response, autoimmune disorder, liver disease, infection, neurological disorder, psychiatric disorder, nitric oxide disorder, urea cycle disorder, congestion, vasodilation disorder, skin disease, wound healing, reaction to insect bites, eye disorder, connective tissue disorder, and infection with certain viruses, bacteria, or fungi. (Item 31) The method according to any of the preceding items, wherein administration of an effective amount of the preparation promotes endothelial function. (Item 32) The method according to any of the preceding items, wherein administration of an effective amount of the preparation alters or changes the level of nitrite or NO in the target tissue or in the circulation. (Item 33) The method according to any of the preceding items, wherein administration of an effective amount of the preparation modulates the microbiome associated with the digestive system of the subject. (Item 34) A method of any of the preceding items, wherein the administration is performed using a device. (Item 35) The method according to any of the preceding items, wherein the preparation is administered before the onset of the gastrointestinal condition. (Item 36) The method according to any of the preceding items, wherein the preparation is administered during the development of a gastrointestinal condition. (Item 37) The method according to any of the preceding items, wherein the preparation is administered after sedation of the gastrointestinal condition. (Item 38) The method according to any of the preceding items, wherein the preparation is administered in response to gastrointestinal symptoms, triggers, or prodromal symptoms, such as discomfort or changes in bowel habits. (Item 39) A method for any of the preceding items, further comprising determining whether the subject requires treatment for a digestive disorder. (Item 40) The method according to any of the preceding items, wherein the preparation is administered as a solution, suspension, emulsion, ointment, gel, hydrogel, or liquid, for example, as a drop, spray, aerosol, or mist. (Item 41) The method according to any of the preceding items, wherein the preparation is formulated as a tablet or capsule. (Item 42) The method according to any of the preceding items, wherein the preparation comprises microspheres or microcapsules. (Item 43) The method according to any of the preceding items, wherein the preparation is formulated to be suitable for the digestive system of the subject. (Item 44) The method according to any of the preceding items, wherein administration reduces bloating, diarrhea, gas, stomach pain, stomach cramps, or borborygmus in the subject. (Item 45) The method of any of the preceding items, wherein the preparation is formulated for immediate release or extended release. (Item 46) The method according to any of the preceding items, wherein the preparation is formulated to deliver nitrite or NO to a target tissue, either locally or systemically. (Item 47) The method according to any of the preceding items, wherein the preparation is formulated, for example, locally or systemically, for transmucosal delivery and / or into circulation. (Item 48) The method of any of the preceding items, further comprising administering a second amount of the preparation to the subject. (Item 49) The method according to any of the preceding items, wherein the preparation is administered as part of a combination therapy. (Item 50) The method of any of the preceding items, further comprising administering a second treatment in combination with the preparation. (Item 51) The method of any of the preceding items, wherein the preparation is administered over a period of time prior to initiating the second treatment. (Item 52) The method of any of the preceding items, wherein the preparation is administered simultaneously with the second treatment. (Item 53) The method according to any of the preceding items, wherein the preparation is administered over a certain period of time after discontinuation of the second treatment. (Item 54) The method of any of the preceding items, wherein the second procedure is administered via an alternative mode of administration, such as inhalation or an intranasal technique. (Item 55) The method described above for any of the preceding items, wherein the subject receives a second treatment at a therapeutic level. (Item 56) The method according to any of the preceding items, wherein the preparation is administered together with an anti-inflammatory agent. (Item 57) The method of any of the preceding items, wherein the preparation is administered in conjunction with a medical approach that is approved to treat or commonly used to treat, for example, a related disease or disorder, or symptoms of a related disease or disorder. (Item 58) The method according to any of the preceding items, wherein the preparation is administered before or after a surgical or diagnostic procedure. (Item 59) The method according to any of the preceding items, wherein the preparation is administered in conjunction with exercise, fiber, laxatives, antidiuretics, probiotics, therapeutic agents, or stress management. (Item 60) The method according to any of the preceding items, wherein the preparation is administered in combination with therapeutic treatment for colitis, necrotizing enterocolitis, inflammatory bowel disease, ulcer, Crohn's disease, ulcerative colitis, celiac disease, gluten sensitivity, heartburn, pancreatitis, appendicitis, gastritis, gastroenteritis, or irritable bowel syndrome. (Item 61) The method according to any of the preceding items, wherein the preparation is administered together with nitrite, nitrate, and / or NO. (Item 62) The method according to any of the preceding items, wherein the effective dose is the therapeutic effective dose for AOM. (Item 63) The aforementioned effective therapeutic dose of AOM is approximately 1 × 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 , or 10 14 A method that is either CFU or a method that exceeds CFU, or any of the preceding items. (Item 64) The method according to any of the preceding items, wherein the preparation is administered as an analgesic. (Item 65) The method according to any of the preceding items, wherein the preparation is administered as a prophylactic agent. (Item 66) The method according to any of the preceding items, wherein the preparation is self-administered. (Item 67) The method according to any of the preceding items, wherein the preparation is administered approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 times per day. (Item 68) The method according to any of the preceding items, wherein the preparation is administered over a period of approximately 1-3, 3-5, 5-7, 7-9, 5-10, 10-14, 12-18, 12-21, 21-28, 28-35, 35-42, 42-49, 49-56, 46-63, 63-70, 70-77, 77-84, or 84-91 days. (Item 69) The method according to any of the preceding items, wherein the preparation is administered within 30, 60, 90, 120, 150, or 180 minutes after the subject wakes up. (Item 70) The method according to any of the preceding items, wherein the preparation is administered within 30, 60, 90, 120, 150, or 180 minutes before the subject goes to sleep. (Item 71) The method according to any of the preceding items, wherein the preparation is administered within 30, 60, 90, 120, 150, or 180 minutes of the subject having a meal. (Item 72) The method according to any of the preceding items, wherein the preparation is administered 30, 60, 90, 120, 150, or 180 minutes before the subject washes or showers. (Item 73) The method described in any of the preceding items, wherein the subject is female. (Item 74) The method described in any of the preceding items, wherein the subject is male. (Item 75) The method described in any of the preceding items, wherein the subject is characterized as one of the following ethnic / racial groups: Asian, Black or African American, Hispanic or Latino, White, or multi-ethnic. (Item 76) The method according to any of the preceding items, wherein the subject has a disturbed microbiome. (Item 77) The method described in any of the preceding items, wherein the subject is under 1 year old, or between 1 and 5 years old, between 5 and 10 years old, between 10 and 20 years old, between 20 and 30 years old, between 30 and 40 years old, between 40 and 50 years old, between 50 and 60 years old, or over 60 years old. (Item 78) The method according to any of the preceding items, wherein the preparation comprises AOM in a buffer solution, for example, in an aqueous buffer solution. (Item 79) The method according to any of the preceding items, wherein the buffer solution, for example, an aqueous buffer solution, contains disodium phosphate and magnesium chloride, for example, 50 mM Na2HPO4 and 2 mM MgCl2 in water. (Item 80) The method according to any of the preceding items, wherein the buffer solution, for example, an aqueous buffer solution, essentially consists of disodium phosphate and magnesium chloride, for example, 50 mM Na2HPO4 and 2 mM MgCl2 in water. (Item 81) The method according to any of the preceding items, wherein the buffer solution, for example, an aqueous buffer solution, consists of disodium phosphate and magnesium chloride, for example, 50 mM Na2HPO4 and 2 mM MgCl2 in water. (Item 82) The method according to any of the preceding items, wherein the preparation is characterized by a physiological pH level. (Item 83) The method according to any of the preceding items, wherein the preparation further comprises, or is administered concurrently with, a compound that promotes the proliferation or metabolism of AOM, NO production, and / or urease activity. (Item 84) The method according to any of the preceding items, wherein the preparation comprises at least one of ammonia, an ammonium salt, and urea. (Item 85) The method according to any of the preceding items, wherein the preparation includes a controlled-release material, such as a delayed-release material. (Item 86) The method according to any of the preceding items, wherein the preparation further comprises an additive, for example, a pharmaceutically acceptable additive. (Item 87) The method according to any of the preceding items, wherein the additive comprises an absorption enhancer or penetration enhancer, a preservative, an antioxidant, a buffer, a chelating agent, an ion exchanger, a solubilizer, a suspending agent, a thickener, a surfactant, a wetting agent, an isotonic agent, an enzyme inhibitor, or a vehicle for appropriate drug delivery. (Item 88) The method according to any of the preceding items, wherein the preparation comprises a mucosal adhesive. (Item 89) The method according to any of the preceding items, wherein the preparation comprises a disintegrant, a chelating agent, a coating agent, a release-controlled formulation, or a filler. (Item 90) The method according to any of the preceding items, wherein the preparation substantially does not contain other organisms. (Item 91) The aforementioned preparation is approximately 1 × 10 3 From CFU / mL to approximately 1 × 10 14 The method described in any of the preceding items, including AOM between CFU / mL. (Item 92) The aforementioned preparation is approximately 1 × 10 9 From CFU / mL to approximately 10 × 10 9 The method described in any of the preceding items, including AOM between CFU / mL. (Item 93) The method according to any of the preceding items, wherein the AOM includes ammonia-oxidizing bacteria (AOB). (Item 94) A method by which the aforementioned AOM is essentially derived from the AOB, in any of the preceding items. (Item 95) The method according to any of the preceding items, wherein the AOM consists of AOB. (Item 96) The method according to any of the preceding items, wherein the AOM includes Nitrosomonas, Nitrosococcus, Nitrosospira, Nitrosocystis, Nitrosolobus, Nitrosovibrio, and combinations thereof. (Item 97) The method described in any of the preceding items, wherein the AOM is Nitrosomonas eutropha (N. eutropha). (Item 98) The method according to any of the preceding items, wherein the AOM is N. eutropha D23 having ATCC accession number PTA-121157. (Item 99) The method according to any of the preceding items, wherein the AOM includes ammonia-oxidizing archaea (AOA). (Item 100) The method according to any of the preceding items, wherein the AOM can convert ammonia or ammonium to nitrite at a rate of at least about 1 pmol / min / mg of protein, for example, at least about 0.1 nmol / min / mg of protein. (Item 101) The method according to any of the preceding items, wherein the preparation is administered, for example, by ingestion into a first tissue, such as a deposited tissue. (Item 102) The method according to any of the preceding items, wherein the first organization is the target organization. (Item 103) The method according to any of the preceding items, wherein the first tissue is other than the target tissue, and for example, the preparation is applied to the first tissue, and the preparation or a product of the preparation, such as NO, is transported to a second tissue, such as the target tissue, for example, by diffusion. (Item 104) A method for any of the preceding items, wherein the second procedure includes a surgical procedure. (Item 105) The method according to any of the preceding items, wherein the additive includes an anti-adhesive agent, a binder, a coating agent, a disintegrant, a filler, a fragrance, a colorant, a lubricant, a flow promoter, an adsorbent, a preservative, or a sweetener. (Item 106) A method according to any of the preceding items, wherein a biome-safe product is used in conjunction with an administered preparation containing an AOM. (Item 107) A preparation containing the AOM described in any of the preceding items, for enteral administration to the target subject. (Item 108) The preparation described in any of the preceding items, wherein the preparation is a food product. (Item 109) The preparation described in any of the preceding items, wherein the food product includes food, beverage, nutritional supplement, functional food, additive, or medical nutrition product. (Item 110) A preparation containing AOM as described in any of the preceding items, for treating gastrointestinal disorders in the subject. (Item 111) The preparation described in any of the preceding items, wherein the preparation is packaged for single use. (Item 112) The preparation described in any of the preceding items, wherein the preparation is packaged for multiple uses. (Item 113) Preparations described in any of the preceding items, including AOM and other organisms, such as biological communities. (Item 114) A device configured to administer a preparation containing AOM as described in any of the preceding items to a target tissue of the gastrointestinal system. (Item 115) A kit containing a preparation containing the AOM described in any of the preceding items.

Claims

[Claim 1] The method described in the specification.