Methods and uses of microbiome compositions, components or metabolites for treating insulin-related diseases

Abstract

Methods and uses of compositions (e.g., comprising one or more microbial strains, one or more components, one or more metabolites, or a combination thereof) for the treatment of insulin-related diseases, disorders, and conditions are disclosed.

Description

[0001] Cross - reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 407,592, filed on September 16, 2022, the entire content of which is hereby incorporated by reference in its entirety. Background of the Invention

[0003] Many insulin - related diseases, disorders or afflictions, including but not limited to diabetes, obesity, cardiovascular diseases, non - alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadromes, diabetic neuropathy, diabetic foot ulcers, maturity - onset diabetes of the young, pancreatogenic diabetes, polycystic ovary syndrome (PCOS) or Alzheimer's disease (AD), can lead to the degeneration of various cells (such as liver cells, pancreatic cells, etc.) and affect physical and / or mental functions. Currently, there is no effective treatment for such diseases, and finding new drugs or treatment methods is an urgent priority. Summary of the Invention

[0004] The present disclosure provides the insight that compositions (such as microbiome compositions) as described herein can be used to treat diseases, disorders or afflictions (such as insulin-related diseases, disorders or afflictions (such as diabetes, obesity, cardiovascular diseases, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, AD, etc.)) in a subject (such as a mammal (such as a human, mouse, etc.)). The present disclosure particularly describes techniques that can be used to treat, prevent diseases, disorders or afflictions (such as insulin-related) and / or reduce their risk. In some embodiments, the present disclosure describes compositions and methods to evaluate the effects of administering such compositions (such as, for example, the microbiome compositions as described herein) to a subject and / or to identify or characterize the effects and / or modulation of levels of metabolites or metabolome in a subject after administering such compositions. In some embodiments, the metabolites that can be modulated may be associated with certain diseases, disorders or afflictions. In some embodiments, such techniques can be used to discern differences in metabolite levels in a particular subject (such as a patient) or population (for example, before and after administering the disclosed compositions). Thus, the present disclosure also provides techniques that can be used to identify and / or evaluate the properties and effects of the disclosed compositions in a particular subject (such as a patient) and / or population and thus provide subject-specific information on how to treat the diseases, disorders or afflictions (such as insulin-related diseases, disorders or afflictions) of an individual subject or individual population. For example, in some embodiments, the techniques provided herein can be used to identify a subject-specific composition based on the metabolome in a subject-specific sample and to treat and / or prevent diseases, disorders or afflictions (such as insulin-related diseases, disorders or afflictions) by administering the disclosed compositions (such as subject-specific compositions) (such as to modulate the metabolome of the subject). Thus, the techniques described herein can be used as therapeutic agents and tools for reducing the risk of certain diseases, disorders or afflictions (such as insulin-related diseases, disorders or afflictions) and for treating and / or preventing such diseases, disorders or afflictions.

[0005] The present disclosure particularly provides a method for treating or preventing insulin-related diseases, disorders or afflictions. In some embodiments, the method comprises administering to a subject a composition comprising one or more microbial strains or microbial components. In some embodiments, the method comprises administering to a subject a composition comprising one or more microbial metabolites. In some embodiments, the method comprises administering to a subject a composition comprising: (i) one or more microbial strains or microbial components, or (ii) one or more microbial metabolites. In some embodiments, the insulin-related diseases, disorders or afflictions are or include diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermopathy, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes or polycystic ovary syndrome (PCOS). In some embodiments, the insulin-related diseases, disorders or afflictions are or include diabetes.

[0006] In some embodiments, the subject has been diagnosed with an insulin-related disease, disorder or affliction or is at high risk of developing an insulin-related disease, disorder or affliction. In some embodiments, the subject is an animal. In some embodiments, the subject is a mammal, such as a mammal that experiences or is susceptible to the diseases, disorders or afflictions described herein. In some embodiments, the animal is a vertebrate, such as a mammal, such as a non-human primate (especially a higher primate), sheep, dog, rodent (e.g., mouse or rat), guinea pig, goat, pig, cat, rabbit or cow. In some embodiments, the animal is a non-mammal, such as a chicken, amphibian, reptile or invertebrate. In some embodiments, the subject is a human.

[0007] In some embodiments, the subject has or is susceptible to one or more insulin-related diseases, disorders or afflictions described herein. In some embodiments, the subject exhibits one or more symptoms of one or more insulin-related diseases, disorders or afflictions described herein. In some embodiments, the subject has been diagnosed with one or more insulin-related diseases, disorders or afflictions described herein. In some embodiments, the subject is receiving or has received a certain therapy for diagnosing and / or treating one or more insulin-related diseases, disorders or afflictions.

[0008] In some embodiments, the one or more microbial strains are from the mammalian microbiome. In some embodiments, the one or more microbial strains are from the human microbiome. In some embodiments, the human microbiome is the microbiome of the subject. In some embodiments, the human microbiome is administered to maintain or regulate the microbiome of the subject.

[0009] In some embodiments, one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3, or Appendix 4. In some embodiments, the metabolite can be from one or more microbial strains. In some embodiments, the metabolite can be from a source other than a microbial strain, such as synthetically produced. In some embodiments, one or more microbial metabolites are or comprise bile acids. In some embodiments, one or more microbial metabolites are or comprise tauroursodeoxycholic acid. In some embodiments, one or more microbial components or microbial metabolites are butyrylcarnitine, theobromine, 4-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof. In some embodiments, one or more microbial components or microbial metabolites are 4-hydroxyphenylpyruvic acid, ectoine, gramine, N-acetyl-L-phenylalanine, Nε-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine, b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenylacetylglycine, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indolylsulfate, nicotinamide, N-formylglycine, ureidoethanolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenylacetylglycine, diethanolamine, phosphocholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiazolidine-4-carboxylic acid, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0XA0027, or any combination thereof.

[0010] In some embodiments, one or more microbial strains are or include Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof. In some embodiments, one or more microbial strains are or include Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof. In some embodiments, one or more microbial strains are or include Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof. In some embodiments, one or more microbial strains are or include Bacillus subtilis.

[0011] In some embodiments, the composition comprises two or more microbial strains. In some embodiments, the composition comprises five or more microbial strains. In some embodiments, the composition comprises ten or more microbial strains.

[0012] In some embodiments, the composition is administered locally, orally, subcutaneously, intravenously, intramuscularly, intracranially, intrathecally, rectally, ophthalmically, intravitreally, or suprachoroidally. In some embodiments, the composition is administered orally. In some embodiments, the composition is administered intravenously.

[0013] In some embodiments, the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection, or eye drop.

[0014] In some embodiments, each microbial strain among one or more microbial strains is present in the composition at a concentration of 10 1 to 10 15 CFU. In some embodiments, each microbial strain among one or more microbial strains is present in the composition at a concentration of at least 10 6The concentration of CFU is present in the composition. In some embodiments, each microbial strain among one or more microbial strains in the composition comprises 10 1 colony forming units (CFU) to 10 20 CFU. In some embodiments, each microbial strain among one or more microbial strains in the composition comprises 10 1 colony forming units (CFU) to 10 15 CFU. In some embodiments, each microbial strain among one or more microbial strains in the composition comprises 10 6 CFU to 10 15 CFU. In some embodiments, each microbial strain among one or more microbial strains in the composition comprises about 10 1 CFU to 10 15 CFU, or about 10 2 CFU to 10 14 CFU, or about 10 3 CFU to 10 13 CFU, or about 10 4 CFU to 10 13 CFU, or about 10 5 CFU to 10 12 CFU, or about 10 6 CFU to 10 11 CFU, or about 10 7 CFU to 10 10 CFU, or about 10 8 CFU to 10 9 CFU, or about 10 5 CFU to 10 10 CFU, or about 10 8 CFU to 10 12 CFU. In some embodiments, each microbial strain among one or more microbial strains in the composition comprises at least about 10 1 , 5x 10 1 , 10 2 , 5x 10 2 , 10 3 , 5x 10 3 , 10 4 , 5x 10 4 , 10 5 , 5x 10 5 , 10 6 , 5x 10 6 , 10 7 , 5x 10 7 , 10 8 , 5x 10 8, 10 9 , 5 x 10 9 , 10 10 , 5 x 10 10 , 10 11 , 5 x 10 11 , 10 12 or more CFU. In some embodiments, each of one or more microbial strains in the composition comprises at most about 10 15 , 5 x 10 14 , 10 14 , 5 x 10 13 , 10 13 , 5 x 10 12 , 10 12 , 5 x 10 11 , 10 11 , 5 x 10 10 , 10 10 , 5 x 10 9 , 10 9 , 5 x 10 8 , 10 8 or fewer CFU. In some embodiments, each microbial strain among one or more microbial strains in the composition comprises the same number of CFU. In some embodiments, some microbial strains among one or more microbial strains in the composition comprise different numbers of CFU.

[0015] The present disclosure particularly provides a composition for treating insulin-related diseases, disorders or afflictions or for use in treating insulin-related diseases, disorders or afflictions, the composition comprising one or more microbial strains, their microbial components or their microbial metabolites. In some embodiments, the composition as described herein comprises one or more microbial metabolites (e.g., derived from sources other than microbial strains (e.g., synthetic sources), derived from one or more microbial strains), wherein the composition is for treating insulin-related diseases, disorders or afflictions.

[0016] The present disclosure provides a composition, the composition comprising one or more microbial strains, the one or more microbial strains being selected from Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Clostridium butyricum, a species of Paenibacillus, a species of Veillonella, a species of Bifidobacterium, Bacillus subtilis, a species of Acidaminococcus, or a combination thereof. In some embodiments, the composition comprises one or more microbial strains, the one or more microbial strains being selected from Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Veillonella atypica, a species of Bifidobacterium, or a combination thereof. In some embodiments, the composition comprises a microbial strain. In some embodiments, the microbial strain is Bacillus subtilis. In some embodiments, the composition comprises at least two microbial strains selected from the group consisting of Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Clostridium butyricum, a species of Paenibacillus, a species of Veillonella, a species of Bifidobacterium, Bacillus subtilis, a species of Acidaminococcus, or a combination thereof. In some embodiments, the composition comprises at least two microbial strains selected from the group consisting of Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Veillonella atypica, a species of Bifidobacterium, or a combination thereof. In some embodiments, the composition comprises at least five microbial strains selected from the group consisting of Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Clostridium butyricum, a species of Paenibacillus, a species of Veillonella, a species of Bifidobacterium, Bacillus subtilis, a species of Acidaminococcus, or a combination thereof. In some embodiments, the composition comprises at least five microbial strains selected from the group consisting of Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Veillonella atypica, a species of Bifidobacterium, or a combination thereof. In some embodiments, the composition comprises or consists of Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Clostridium butyricum, a species of Paenibacillus, a species of Veillonella, a species of Bifidobacterium, Bacillus subtilis, a species of Acidaminococcus. In some embodiments, the composition comprises or consists of Acetobacter hansenii, Terribacillus glycolicus, a species of Coprococcus, Lactobacillus plantarum, Veillonella atypica, a species of Bifidobacterium.

[0017] In some embodiments, the composition as described herein comprises one or more microbial metabolites (e.g., derived from sources other than the microbial strains (e.g., synthetic sources), derived from one or more microbial strains), wherein the composition is for treating insulin-related diseases, disorders, or afflictions.

[0018] In some embodiments, the composition is for topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal, ocular, intravitreal or suprachoroidal administration. In some embodiments, the composition is for oral administration. In some embodiments, the composition is for intravenous administration.

[0019] The present disclosure particularly provides a composition for treating insulin-related diseases, disorders or conditions, the composition comprising one or more microbial strains or microbial components. In some embodiments, the composition for treating insulin-related diseases, disorders or conditions comprises one or more microbial metabolites.

[0020] The present disclosure provides the composition as described herein for modulating one or more microbial metabolites in a subject. In some embodiments, the composition is for modulating one or more characteristics of a subject. In some embodiments, the one or more characteristics are or include (i) the level of cell viability; (ii) the level or activity of nucleic acids or proteins, or forms thereof; (iii) weight gain; (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride level; (vii) blood cholesterol level; (viii) oxidative stress; or (ix) inflammation.

[0021] In some embodiments, the composition is for characterizing the ability of one or more microbial strains to modulate one or more microbial metabolites in a subject.

[0022] The present disclosure provides the use of the composition as described herein for treating or improving a disease, disorder or condition of a subject, wherein the disease, disorder or condition is an insulin-related disease, disorder or condition associated with one or more microbial metabolites. In some embodiments, the use of the composition as described herein is for treating or improving diabetes.

[0023] The present disclosure provides a method for screening a microbial strain, the method comprising contacting the microbial strain with a culture comprising pancreatic cells or a pancreatic cell line that mimics an insulin-related disease, disorder or condition, and determining whether the microbial strain has altered a characteristic of the culture, wherein the characteristic is associated with the insulin-related disease, disorder or condition.

[0024] In some embodiments, the determining step comprises comparing the characteristics before and after performing the contacting step. In some embodiments, the determining step comprises comparing the characteristics after the contacting step with a comparable reference.

[0025] In some embodiments, the comparable reference is a historical reference. In some embodiments, the comparable reference is a negative control reference. In some embodiments, the comparable reference is a positive control reference.

[0026] In some embodiments, the feature is the level of cell viability. In some embodiments, the feature is the level or activity of a nucleic acid or protein, or a form thereof. In some embodiments, the feature is or includes weight gain. In some embodiments, the feature is or includes fat accumulation in hepatocytes. In some embodiments, the feature is or includes lipid accumulation in hepatocytes. In some embodiments, the feature is or includes triglyceride level. In some embodiments, the feature is or includes cholesterol level. In some embodiments, the feature is or includes inflammation.

[0027] In some embodiments, a microbial strain can alter one or more features of a culture. In some embodiments, one or more features are related to an insulin-related disease, disorder, or condition. In some embodiments, one or more features are or include (i) the level of cell viability; (ii) the level or activity of a nucleic acid or protein, or a form thereof; (iii) weight gain; (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride level; (vii) blood cholesterol level; (viii) oxidative stress; or (ix) inflammation.

[0028] The present disclosure provides a method that includes administering to a subject a composition comprising one or more microbial strains or microbial components. In some embodiments, the present disclosure provides a method that includes administering to a subject a composition comprising one or more microbial metabolites. In some embodiments, the microbial metabolites can be from one or more microbial strains. In some embodiments, the microbial metabolites can be from a non-microbial strain source, such as synthetically produced.

[0029] In some embodiments, a microbial strain, microbial component, or microbial metabolite can alter a feature of a subject. In some embodiments, a microbial strain, microbial component, or microbial metabolite can alter one or more features of a subject. In some embodiments, the feature is the level of cell viability. In some embodiments, the feature is the level or activity of a nucleic acid or protein, or a form thereof. In some embodiments, the feature is or includes weight gain. In some embodiments, the feature is or includes fat accumulation in the liver. In some embodiments, the feature is or includes lipid accumulation in the liver. In some embodiments, the feature is or includes triglyceride level. In some embodiments, the feature is or includes cholesterol level. In some embodiments, the feature is or includes inflammation.

[0030] In some embodiments, the feature is related to an insulin-related disease, disorder, or condition.

[0031] The present disclosure provides a method for characterizing a microbial strain, the method comprising adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that mimic an insulin-related disease, disorder, or condition, and determining whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are related to the insulin-related disease, disorder, or condition.

[0032] The present disclosure provides a method for manufacturing a pharmaceutical treatment, the method comprising characterizing one or more microbial strains, microbial components, or microbial metabolites, comprising the steps of: adding the one or more microbial strains to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that mimic an insulin-related disease, disorder, or condition, and determining whether the one or more microbial strains affect the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are related to the insulin-related disease, disorder, or condition.

[0033] The present disclosure provides a method for manufacturing a pharmaceutical treatment, the method comprising formulating one or more microbial strains or microbial compositions in a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection, or eye drop. The present disclosure provides a method for manufacturing a pharmaceutical treatment, the method comprising formulating one or more microbial metabolites in a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection, or eye drop.

[0034] The present disclosure provides a method for assessing the ability of a microbial strain to affect one or more characteristics of a culture, the method comprising adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that mimic an insulin-related disease, disorder, or condition, and determining whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are related to the insulin-related disease, disorder, or condition.

[0035] In some embodiments, the method further comprises, prior to adding the microbial strain to the culture, determining the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture, after adding the microbial strain to the culture, determining the level of the same one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture, and comparing the level of the one or more characteristics determined prior to adding the microbial strain with the level of the one or more characteristics determined after adding the microbial strain.

[0036] In some embodiments, one or more features include (i) the level of cell viability; (ii) the level or activity of a nucleic acid or protein, or a form thereof; (iii) weight gain; (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride level; (vii) blood cholesterol level; (viii) oxidative stress; (ix) inflammation; or (x) a combination thereof.

[0037] The present disclosure provides a composition as described herein for treating or preventing an insulin-related disease, disorder, or condition, the composition comprising one or more microbial strains or microbial components. The present disclosure further provides a composition as described herein for treating or preventing an insulin-related disease, disorder, or condition, the composition comprising one or more microbial metabolites. In some embodiments, one or more microbial components or microbial metabolites of one or more microbial strains are selected from Appendix 1, Appendix 3, or Appendix 4.

[0038] In some embodiments, a composition as described herein is used for treating or preventing an insulin-related disease, disorder, or condition, and the insulin-related disease, disorder, or condition is diabetes.

[0039] The present disclosure provides an injection comprising a composition as described herein.

[0040] The present disclosure provides a food supplement comprising a composition as described herein.

[0041] The present disclosure provides a kit comprising a composition as described herein, the kit being for treating or preventing an insulin-related disease, disorder, or condition. In some embodiments, the kit comprises a monitoring device. In some embodiments, the monitoring device is a blood glucose monitor.

[0042] These and other aspects covered by the present disclosure are described in more detail below and in the claims.

[0043] Definitions

[0044] The scope of the present invention is defined by the appended claims and is not limited by certain embodiments described herein. Those skilled in the art reading this specification will recognize various modifications that are equivalent to such described embodiments or otherwise within the scope of the claims. Generally, unless otherwise clearly indicated, the terms used herein have the meanings as understood in the art. Defined meanings of certain terms are provided below; throughout this specification, the meanings of these and other terms will be apparent to those skilled in the art from the context in particular instances.

[0045] The use of sequential terms such as “first,” “second,” “third,” etc. in the claims to modify the claim elements themselves does not mean any priority, precedence, or order of one claim element with respect to another claim element or the chronological order of performing method acts, but is used only as a label to distinguish one claim element having a particular name from another element having the same name (but using the sequential terms) to distinguish the claim elements.

[0046] Unless expressly indicated to the contrary, as used herein, the articles “a / an” shall be understood to include plural referents. Unless indicated to the contrary or otherwise apparent from the context, a claim or description that includes “or” among one or more members of a group is satisfied if one, more than one, or all of the members of the group are present in, employed in, or otherwise involved in a given product or process. In some embodiments, only one member of the group is present in, employed in, or otherwise involved in a given product or process. In some embodiments, more than one or all of the members of the group are present in, employed in, or otherwise involved in a given product or process. It should be understood that, unless otherwise indicated or unless it would be apparent to one of ordinary skill in the art that a contradiction or inconsistency would arise, the present invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, descriptive terms, etc. from one or more of the listed claims are introduced into another claim that depends from the same independent claim (or any other relevant claim). In cases where elements are presented in a list (e.g., in a Markush group or similar format), it should be understood that subgroups of the elements are also disclosed and any element may be removed from the group. It should be understood that, generally speaking, where an embodiment or aspect is referred to as “comprising” a particular element, feature, etc., certain embodiments or aspects “consist of” or “consist essentially of” such element, feature, etc. For simplicity, those embodiments are not specifically set forth in so many words in each instance herein. It should also be understood that any embodiment or aspect may be expressly excluded from the claims, whether or not a particular exclusion is recited in the specification.

[0047] Administration: As used herein, the term "administration" generally refers to the administration of a composition to a subject or system to effect delivery of an agent to the subject or system. In some embodiments, the agent is the composition or is included in the composition; in some embodiments, the agent is produced by metabolism of the composition or one or more of its components. Those of ordinary skill in the art will appreciate the various routes that can be used to administer to a subject (e.g., a human) as appropriate. For example, in some embodiments, administration can be ocular, oral, parenteral, topical, etc. In some specific embodiments, administration can be bronchial (e.g., by bronchial instillation), buccal, cutaneous (which can be or include, for example, topical application to one or more of the dermis, intradermal, intercutaneous, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a particular organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, etc. In many of the embodiments provided by the present disclosure, administration is oral administration. In some embodiments, administration can involve only a single dose. In some embodiments, administration can involve the application of a fixed number of doses. In some embodiments, administration can involve intermittent (e.g., multiple doses separated in time) and / or periodic (e.g., individual doses separated by the same time period) dosing. In some embodiments, administration can involve continuous dosing (e.g., perfusion) for at least a selected period of time. Administration of cells can be carried out by any suitable route that effects delivery to the desired location in the subject, where at least a portion of the delivered cells or components of the cells remain viable. The survival period of the cells after administration to the subject can be as short as a few hours, e.g., twenty-four hours, to several days, to up to several years, i.e., long-term implantation. In some embodiments, administration includes the delivery of a bacterial extract or preparation that contains one or more bacterial metabolites and / or by-products but lacks fully viable bacterial cells.

[0048] Analogue: As used herein, the term "analogue" refers to a substance that shares one or more specific structural features, elements, components, or moieties with a reference substance. Generally, an "analogue" exhibits significant structural similarity to the reference substance (e.g., shares a core or common structure) but also differs in certain discrete ways. In some embodiments, an analogue is a substance that can be produced, for example, from a reference substance by chemical manipulation of the reference substance. In some embodiments, an analogue is a substance that can be produced by performing a synthetic process that is substantially similar to (e.g., shares multiple steps with) the synthetic process used to produce the reference substance. In some embodiments, an analogue can be produced by performing a synthetic process that is different from the synthetic process used to produce the reference substance.

[0049] About: When applied to one or more values of interest, includes values similar to the recited reference values. In certain embodiments, unless otherwise stated or otherwise apparent from the context (except in cases where such numerical values would exceed 100% of the possible values), the term “about” or “approximately” refers to a range of values that fall within (greater than or less than) ±10% of the recited reference value.

[0050] Comparable: As used herein, the term “comparable” refers to two or more agents, entities, situations, groups of conditions, subjects, etc. that may not be identical to each other but are sufficiently similar to permit comparison between them such that one of ordinary skill in the art would understand that conclusions can be reasonably drawn based on the observed differences or similarities. In some embodiments, groups of comparable conditions, environments, individuals, or populations are characterized by a plurality of substantially identical characteristics and one or a small number of different characteristics. One of ordinary skill in the art will understand, in context, what degree of congruence is required for two or more such agents, entities, situations, groups of conditions, etc. to be considered comparable in any given case. For example, one of ordinary skill in the art will understand that environments, individuals, or groups of populations are comparable to each other when characterized by a sufficient number and type of substantially identical characteristics to warrant a reasonable conclusion that differences in results or observed phenomena obtained in the case of different environments, individuals, or groups of populations are caused or indicated by variations in those varying characteristics.

[0051] Conservative: As used herein, refers to the situation when describing conservative amino acid substitutions, including the replacement of an amino acid residue with another amino acid residue having a side chain R group with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially change the functional properties of interest of a protein, e.g., the ability of a receptor to bind to a ligand. Examples of groups of amino acids having side chains with similar chemical properties include: aliphatic side chains such as glycine (Gly, G), alanine (Ala, A), valine (Val, V), leucine (Leu, L), and isoleucine (Ile, I); aliphatic hydroxyl side chains such as serine (Ser, S) and threonine (Thr, T); amide-containing side chains such as asparagine (Asn, N) and glutamine (Gln, Q); aromatic side chains such as phenylalanine (Phe, F), tyrosine (Tyr, Y), and tryptophan (Trp, W); basic side chains such as lysine (Lys, K), arginine (Arg, R), and histidine (His, H); acidic side chains such as aspartic acid (Asp, D) and glutamic acid (Glu, E); and sulfur-containing side chains such as cysteine (Cys, C) and methionine (Met, M). Groups of conservative amino acid substitutions include, for example, valine / leucine / isoleucine (Val / Leu / Ile, V / L / I), phenylalanine / tyrosine (Phe / Tyr, F / Y), lysine / arginine (Lys / Arg, K / R), alanine / valine (Ala / Val, A / V), glutamic acid / aspartic acid (Glu / Asp, E / D), and asparagine / glutamine (Asn / Gln, N / Q). In some embodiments, a conservative amino acid substitution can be the replacement of any native residue in a protein with alanine, e.g., for alanine-scanning mutagenesis. In some embodiments, conservative substitutions are made that have a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet, G.H. et al., 1992, Science 256:1443-1445, which is incorporated herein by reference in its entirety. In some embodiments, the substitution is a moderately conservative substitution, where the substitution has a non-negative value in the PAM250 log-likelihood matrix.

[0052]

[0053]

[0054] Control: As used herein, "control" refers to the meaning understood in the art as the standard for comparison of results. Generally, controls are used to enhance the integrity of experiments by isolating variables to draw conclusions about such variables. In some embodiments, a control is a reaction or assay that is run concurrently with a test reaction or assay to provide a comparator. "Control" also includes "control animal." A "control animal" may have the modifications described herein, different modifications than those described herein, or no modifications (i.e., a wild-type animal). In one experiment, a "test" (i.e., the variable being tested) is applied. In a second experiment, no "control," i.e., the variable being tested, is applied. In some embodiments, a control is a historical control (i.e., a previously run test or assay, or a previously known quantity or result). In some embodiments, a control is or includes a printed or otherwise preserved record. A control can be a positive control or a negative control.

[0055] Determine, measure, evaluate, assess, assay, and analyze: Determine, measure, evaluate, assess, assay, and analyze are used interchangeably herein to refer to any form of measurement and include determining whether an element is present. These terms include quantitative and / or qualitative determinations. An assay can be relative or absolute. "Assaying for presence" can be determining the amount of something present and / or determining whether it is present.

[0056] Dosage form: Those skilled in the art will understand that the term "dosage form" can be used to refer to a physically discrete unit of an agent (e.g., a therapeutic agent) for administration to a subject. Generally, each such unit contains a predetermined amount of the agent. In some embodiments, such amount is an amount that is a unit dose (or a whole part thereof) suitable for administration according to a dosing regimen that has been determined to be relevant to the desired or beneficial outcome (i.e., related to a therapeutic dosing regimen) when administered to a relevant population. Those of ordinary skill in the art understand that the total amount of a therapeutic composition or agent administered to a particular subject is determined by one or more attending physicians and may involve the administration of multiple dosage forms.

[0057] Dosing regimen: Those skilled in the art will understand that the term "dosing regimen" can be used to refer to a set of unit doses (usually more than one) that are typically administered to a subject separately at intervals of time. In some embodiments, a given agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, the dosing regimen includes multiple doses, each dose being separated from the other doses in time. In some embodiments, the individual doses are separated from each other by the same length of time period; in some embodiments, the dosing regimen includes multiple doses and at least two different time periods separating the individual doses. In some embodiments, all the doses within the dosing regimen have the same unit dose amount. In some embodiments, the different doses within the dosing regimen have different amounts. In some embodiments, the dosing regimen includes a first dose of a first dose amount, followed by one or more additional doses of a second dose amount different from the first dose amount. In some embodiments, the dosing regimen includes a first dose of a first dosing amount, followed by one or more other doses of a second dosing amount that is the same as the first dosing amount. In some embodiments, when administered across a relevant population, the dosing regimen is associated with a desired or beneficial outcome.

[0058] Engineering: Generally speaking, the term "engineering" refers to aspects that have been manually manipulated by humans. For example, if a cell or organism has been manipulated to change its genetic information (e.g., new genetic material that did not previously exist has been introduced by, for example, transformation, mating, somatic hybridization, transfection, transduction, or other mechanisms, or existing genetic material has been altered or removed by, for example, substitution or deletion mutations or by mating schemes), then the cell or organism is considered "engineered". By convention and as understood by those skilled in the art, the progeny of engineered polynucleotides or cells are generally still referred to as "engineered", even if the actual manipulation was performed on a previous entity.

[0059] Excipient: As used herein, refers to an inactive (e.g., non-therapeutic) agent that can be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect. In some embodiments, suitable pharmaceutical excipients can include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, ethylene glycol, water, ethanol, etc.

[0060] Functional: As used herein, a "functional" biomolecule is a biomolecule in a form that exhibits properties and / or activities by which it can be characterized. A biomolecule can have two functions (i.e., bifunctional) or multiple functions (i.e., multifunctional).

[0061] Gene: As used herein, refers to a DNA sequence in a chromosome that encodes a product (e.g., an RNA product and / or a polypeptide product). In some embodiments, a gene includes a coding sequence (i.e., a sequence that encodes a specific product). In some embodiments, a gene includes non-coding sequences. In certain specific embodiments, a gene may include both coding (e.g., exons) and non-coding (e.g., introns) sequences. In some embodiments, a gene may include one or more regulatory sequences (e.g., promoters, enhancers, etc.) and / or intron sequences that can control or affect one or more aspects of gene expression (e.g., cell type-specific expression, inducible expression, etc.). For clarity, note that as used in this application, the term "gene" generally refers to a portion of a nucleic acid that encodes a polypeptide or a fragment thereof; the term may optionally encompass regulatory sequences, as will be apparent to one of ordinary skill in the art from the context. This definition is not intended to exclude the application of the term "gene" to non-protein-coding expression units, but rather to clarify that in most cases, the term as used in this document refers to a nucleic acid encoding a polypeptide.

[0062] Improve, increase, enhance, inhibit or decrease: As used herein, the terms "improve", "increase", "enhance", "inhibit", "decrease" or their grammatical equivalents indicate a value relative to a baseline or other reference measurement. In some embodiments, the value has a statistically significant difference from the baseline or other reference measurement. In some embodiments, the appropriate reference measurement may be or include a measurement in a particular system (e.g., in a single individual) under otherwise comparable conditions in the absence / presence of a particular agent or treatment (e.g., before and / or after), or in the presence of an appropriately comparable reference agent. In some embodiments, the appropriate reference measurement may be or include a measurement in a comparable system known or expected to respond in a particular manner in the presence of the relevant agent or treatment. In some embodiments, the appropriate reference is a negative reference; in some embodiments, the appropriate reference is a positive reference.

[0063] Isolated: As used herein, means that a substance and / or entity has (1) been separated from at least some of the components with which it was associated when it was initially produced (whether in nature and / or in an experimental setting), and / or (2) been manually designed, produced, prepared, and / or manufactured by a person. In some embodiments, the isolated substance or entity can be enriched; in some embodiments, the isolated substance or entity can be pure. In some embodiments, the isolated substance and / or entity can be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more than about 99% of the other components with which it was initially associated. In some embodiments, the isolated agent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or greater than about 99% pure. As used herein, a substance is "pure" if it is substantially free of other components. In some embodiments, as will be understood by one of ordinary skill in the art, a substance can still be considered "enriched", "isolated" or even "pure" after being combined with certain other components such as one or more carriers or excipients (e.g., buffers, solvents, water, etc.); in such embodiments, the percentage of separation or purity of the substance is calculated without including such carriers or excipients. One of ordinary skill in the art is aware of various techniques for separating (e.g., enriching or purifying) substances or agents (e.g., using one or more of fractionation, extraction, precipitation, or other separations).

[0064] Level: As used herein, the term "level" refers to a scale of the quantity or amount of a substance (e.g., metabolite). In some embodiments, the level can simply be the presence or absence of the substance. The level of a substance can be expressed in a variety of ways or forms. For example, in some embodiments, the level can be expressed as a percentage (%), a weight measurement (e.g., mg, μg, ng, etc.), a concentration measurement (e.g., mg / mL, μg / mL, ng / mL, etc.), a volume measurement (e.g., mL, μL, nL, etc.), a percent change, etc.

[0065] Metabolite: As used herein, the term "metabolite" refers to a substance (e.g., small molecule, macromolecule, organic compound, or inorganic compound) that is produced or used in a metabolic process. Metabolism is generally understood to be the process by which substances (e.g., food, drugs, chemicals, cells, or tissues) are chemically broken down. In some embodiments, the metabolite is an end product. In some embodiments, the metabolite is an intermediate. Exemplary metabolites are provided herein, for example, in Appendices 1-1, 1-3, and 3. Exemplary metabolic pathways are provided herein, for example, in Appendix 1-2. In some embodiments, the metabolite can be produced or made by an organism. In some embodiments, the metabolite can be produced or made by a microorganism (e.g., a microbial strain). In some embodiments, the microbial metabolite is produced or made by a microbial strain. In some embodiments, the metabolite can be produced or made naturally (e.g., by an organism (e.g., a microorganism (e.g., a microbial strain))). In some embodiments, the metabolite can be produced or made synthetically (e.g., from a source that is not a microbial strain (e.g., synthetically produced)).

[0066] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to a composition in which an active agent is formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose suitable for administration in a treatment regimen that, when administered to a relevant population, shows a statistically significant probability of achieving a predetermined therapeutic effect. In some embodiments, the pharmaceutical composition can be specially formulated for administration in solid or liquid form, including those suitable for: ocular administration, intravitreal administration, suprachoroidal administration, oral administration, subcutaneous administration, intravenous administration, intramuscular administration, intracerebral administration, intrathecal administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets (e.g., those targeted for buccal, sublingual, and systemic absorption), boluses, powders, granules, pastes applied to the tongue, capsules, powders, etc. In some embodiments, the active agent can be or comprise a cell or cell population (e.g., a culture, e.g., a culture of an ellagic acid synthase (EES) microorganism); in some embodiments, the active agent can be or comprise an extract or component of a cell or cell population (e.g., a culture). In some embodiments, the active agent can be or comprise an isolated, purified, or pure compound. In some embodiments, the active agent may have been synthesized in vitro (e.g., by chemical and / or enzymatic synthesis). In some embodiments, the active agent can be or comprise a natural product (whether isolated from its natural source or synthesized in vitro).

[0067] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable" can be used, for example, to refer to a carrier, diluent, or excipient used in formulating a pharmaceutical composition as disclosed herein, meaning that the carrier, diluent, or excipient is compatible with the other components of the composition and harmless to its recipient.

[0068] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle involved in carrying or transporting a subject compound from one organ or part of the body to another organ or part of the body, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject (e.g., the patient). Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository wax; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; diols, such as propylene glycol; polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; pH buffering solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic compatible substances used in pharmaceutical formulations.

[0069] Prebiotic: As used herein, a "prebiotic" is a component that permits or promotes specific changes in the composition and / or activity of the gastrointestinal microbiota that may (or may not) confer a benefit to the host. In some embodiments, the prebiotic can include one or more of the following: Prebiotics include extracts of pome fruits, extracts of berries, and extracts of walnuts.

[0070] Prevention: As used herein, the term "prevention" refers to delaying the onset of a particular disease, disorder, or condition, and / or reducing the frequency and / or severity of one or more symptoms of said disease, disorder, or condition. In some embodiments, prevention is evaluated on a population basis, and thus an agent is considered to "prevent" a disease, disorder, or condition if a statistically significant decrease in the development, frequency, and / or intensity of one or more symptoms of said disease, disorder, or condition is observed in a population susceptible to said disease, disorder, or condition. In some embodiments, prevention can be considered complete, for example, when the onset of a disease, disorder, or condition has been delayed for a predetermined period of time.

[0071] Reference: A standard or control against which comparisons are made as described herein. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially contemporaneously with the target test or determination. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Generally, as will be understood by one of ordinary skill in the art, the reference or control is determined or characterized under conditions or circumstances comparable to the evaluation conditions or circumstances. One of ordinary skill in the art will understand when there is sufficient similarity to justify reliance on and / or comparison to a particular potential reference or control. In some embodiments, the reference is a negative control reference; in some embodiments, the reference is a positive control reference.

[0072] Risk: As will be understood from the context, the "risk" of a disease, disorder, and / or affliction refers to the likelihood that a particular individual will develop the disease, disorder, and / or affliction. In some embodiments, the risk is expressed as a percentage. In some embodiments, the risk is 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or up to 100%. In some embodiments, the risk is expressed as a risk relative to the risk associated with a reference sample or group of reference samples. In some embodiments, the reference sample or group of reference samples has a known risk of a disease, disorder, condition, and / or event. In some embodiments, the reference sample or group of reference samples is from an individual comparable to the particular individual. In some embodiments, the relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or greater.

[0073] Sample: As used herein, the term "sample" generally refers to an aliquot of material obtained or derived from a source of interest. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest can be or include cells or organisms, such as microorganisms, plants or animals (e.g., humans). In some embodiments, the source of interest is or includes biological tissue or fluid. In some embodiments, the biological tissue or fluid can be or include amniotic fluid, aqueous humor, ascites, bile, bone marrow, blood, breast milk, cerebrospinal fluid, cerumen, chyle, chyme, ejaculate, endolymph, exudate, feces, gastric acid, gastric juice, lymph, mucus, pericardial fluid, perilymph, peritoneal fluid, pleural fluid, pus, tenacious mucus, saliva, sebum, semen, serum, smegma, sputum, synovial fluid, sweat, tears, urine, vaginal discharge, vitreous humor, vomitus, plasma, mucus, digestive fluid, feces, and / or combinations or one or more components thereof. In some embodiments, the biological fluid can be or include intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph fluid, and / or transcellular fluid. In some embodiments, the biological fluid can be or include plant secretions. In some embodiments, the biological tissue or sample can be obtained by, for example, aspiration, biopsy (e.g., fine needle or tissue biopsy), swab (e.g., oral, nasal, skin or vaginal swab), scraping, surgery, washing or lavage (e.g., bronchoalveolar, catheter, nasal, eye, oral, uterine, vaginal or other washing or lavage). In some embodiments, the biological sample is or includes cells obtained from an individual. In some embodiments, the sample is an "original sample" obtained directly from the source of interest by any suitable means. In some embodiments, it will be clear from the context that the term "sample" refers to a preparation obtained by processing the original sample (e.g., by removing one or more components and / or by adding one or more agents thereto). By way of example, filtration using a semipermeable membrane. Such a "processed sample" can contain, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the primary sample to one or more techniques, such as amplification or reverse transcription of nucleic acids, separation and / or purification of certain components, etc.

[0074] Small molecule: As used herein, the term "small molecule" refers to a small organic or inorganic molecule having a molecular weight of less than about 3,000 Daltons. Generally, a small molecule may have a molecular weight of less than 3,000 Daltons (Da). A small molecule can be, for example, at least about 100 Da to about 3,000 Da (e.g., about 100 to about 3,000 Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1500 Da, about 500 to about 1000 Da, about 300 to about 1000 Da, or between about 100 and about 250 Da).

[0075] Subject: As used herein, the term "subject" refers to an individual to whom the provided treatment is administered. In some embodiments, the subject is an animal. In some embodiments, the subject is a mammal, such as a mammal that has experienced or is susceptible to a disease, disorder, or condition as described herein. In some embodiments, the animal is a vertebrate, such as a mammal, such as a non-human primate (especially a higher primate), sheep, dog, rodent (e.g., mouse or rat), guinea pig, goat, pig, cat, rabbit, or cow. In some embodiments, the animal is a non-mammal, such as a chicken, amphibian, reptile, or the invertebrate model Caenorhabditis elegans. In some embodiments, the subject is a human. In some embodiments, the subject has or is susceptible to one or more diseases, disorders, or conditions as described herein. In some embodiments, the subject exhibits one or more symptoms of one or more diseases, disorders, or conditions as described herein. In some embodiments, the subject has been diagnosed with one or more diseases, disorders, or conditions as described herein. In some embodiments, the subject is receiving or has received a certain therapy for diagnosing and / or treating a disease, disorder, or condition. In another embodiment, the subject is an experimental animal or animal substitute as a disease model.

[0076] Substantially: As used herein, it refers to a qualitative condition that exhibits the overall or near overall range or degree of the property or characteristic of interest. One of ordinary skill in the biological arts will appreciate that biological and chemical phenomena rarely (if ever) achieve complete and / or proceed to complete or achieve or avoid absolute results. Thus, the term "substantially" is used herein to account for the inherent lack of potential completeness in many biological and chemical phenomena.

[0077] Treatment regimen: The term "treatment regimen" as used herein refers to a dosing schedule whose administration in a relevant population may be associated with a desired or beneficial treatment outcome.

[0078] Therapeutically effective amount: As used herein, refers to the amount that produces the desired effect upon administration. In some embodiments, the term refers to an amount sufficient to treat a disease, disorder, and / or affliction when administered to a population of subjects having or susceptible to the disease, disorder, and / or affliction according to a therapeutic dosing regimen. In some embodiments, a therapeutically effective amount is an amount that reduces the incidence and / or severity of and / or delays the onset of one or more symptoms of a disease, disorder, and / or affliction. One of ordinary skill in the art will understand that the term "therapeutically effective amount" does not actually require successful treatment in a particular individual. Rather, a therapeutically effective amount can be an amount that provides a particular desired pharmacological response in a substantial number of subjects when administered to subjects (e.g., patients) in need of such treatment. In some embodiments, reference to a therapeutically effective amount can be reference to an amount as measured in one or more particular tissues (e.g., tissues affected by a disease, disorder, or affliction) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). One of ordinary skill in the art will understand that in some embodiments, a particular agent or therapy of a therapeutically effective amount can be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective agent can be formulated and / or administered in multiple doses (e.g., as part of a dosing regimen).

[0079] Treatment: As used herein, the term "treatment" (also "treat" or "treating") refers to any administration of a therapy that partially or completely alleviates, ameliorates, relieves, inhibits one or more symptoms, features, and / or causes of a particular disease, disorder, and / or affliction, delays its onset, reduces its severity, and / or reduces its incidence. In some embodiments, such treatment can be directed to subjects who do not exhibit signs of the relevant disease, disorder, and / or affliction and / or subjects who exhibit only early signs of the disease, disorder, and / or affliction. Alternatively or additionally, such treatment can be directed to subjects who exhibit one or more established signs of the relevant disease, disorder, and / or affliction. In some embodiments, treatment can be directed to subjects who have been diagnosed as having the relevant disease, disorder, and / or affliction. In some embodiments, treatment can be directed to subjects known to have one or more risk factors that are statistically associated with an increased risk of development of the relevant disease, disorder, and / or condition. BRIEF DESCRIPTION OF THE DRAWINGS

[0080] Figure 1Graph showing the relative values of IDE to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups are G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m. The graph shows ANOVA comparisons to group G2 (left) and group G1 (right).

[0081] Figure 2 Graph showing the relative values of NSE to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups are G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m.

[0082] Figure 3 Graph showing the relative values of p-Akt Ser473 to actin (left) and Akt1 to actin (right) in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups are G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m.

[0083] Figure 4 Graph showing the relative values of insulin receptor β to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups are G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m.

[0084] Figure 5 Graph showing the relative values of Glut3 to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups are G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m.

[0085] Figure 6 Graph showing the relative values of RBAP48 to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups are G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m.

[0086] Figure 7 Graph showing the relative values of p-4EBP1 to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups were G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m.

[0087] Figure 8 Graph showing the relative values of NRF2 to actin in each of five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. The groups were G1 = wild-type untreated, G2 = transgenic untreated, G3 = transgenic CT10, G4 = transgenic CT10x, G5 = transgenic CT10m. Detailed Description

[0088] Insulin-related diseases, disorders, and afflictions

[0089] Insulin-related diseases, disorders, and afflictions are a collective term for a range of diseases, disorders, and afflictions that are mainly caused by altered or unregulated insulin levels in the human body. Insulin is a hormone that helps the body use glucose from food as an energy source. There are several disease states associated with disorders of insulin secretion and body utilization. Many insulin-related diseases, disorders, or afflictions, including but not limited to diabetes, obesity, cardiovascular diseases, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermopathy, diabetic neuropathy, diabetic foot ulcers, maturity-onset diabetes of the young, pancreatogenic diabetes, polycystic ovary syndrome (PCOS), or Alzheimer's disease (AD), can lead to the degeneration of various cells (such as liver cells, pancreatic cells, etc.) and affect physical and / or mental functions. Some of these are temporary, but others are incurable (e.g., chronic) and debilitating afflictions that result in the progressive degeneration and / or death of cells (such as liver cells, pancreatic cells, etc.), manifested as impaired physical and / or mental functions.

[0090] Insulin levels are unregulated or insulin resistance (also known as impaired insulin sensitivity) occurs when cells in muscle, fat, and the liver do not respond properly to insulin. Muscle, fat, and liver cells may respond inappropriately to insulin, which results in inefficient uptake or storage of glucose from the blood. As a result, the pancreas produces more insulin to try to overcome the rising blood glucose levels. This is called hyperinsulinemia. As long as the pancreas can produce enough insulin to overcome the weak response to insulin, blood glucose levels will remain within a healthy range. If the cells become too resistant to insulin, it can lead to elevated blood glucose levels (hyperglycemia), which over time can lead to prediabetes and type 2 diabetes. In addition to type 2 diabetes, insulin resistance is also associated with several other disorders, including obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, polycystic ovary syndrome (PCOS), and / or other diseases, conditions, or disorders disclosed herein.

[0091] Elevated insulin levels can also lead to weight gain, which in turn makes insulin resistance worse. It is also associated with higher triglyceride levels, hardening of the arteries (atherosclerosis), high blood pressure (hypertension), etc.

[0092] Insulin resistance is also a major feature of metabolic syndrome, which is a cluster of characteristics that link excess fat around the waist and insulin resistance to an increased risk of cardiovascular disease, stroke, and type 2 diabetes. The characteristics of metabolic syndrome include elevated blood glucose levels, elevated triglyceride levels, low high-density lipoprotein (HDL) cholesterol levels, and / or high blood pressure.

[0093] Multiple factors and conditions can lead to varying degrees of insulin resistance. It is believed that excess body fat (especially around the abdomen) and lack of physical activity are two major contributing factors to insulin resistance. In addition to this, insulin resistance may also be caused by diet, certain medications, hormonal imbalances (such as Cushing's syndrome, acromegaly, hypothyroidism, etc.), genetic disorders (such as type A insulin resistance syndrome, Rabson-Mendenhall syndrome, Donohue syndrome, etc.), and other genetic disorders (myotonic dystrophy, Alström syndrome syndrome), Werner syndrome, genetic lipodystrophy, etc.).

[0094] The present disclosure provides compositions (e.g., microbiome compositions) and methods for inhibiting one or more events or processes occurring in insulin-related diseases, disorders, or afflictions. The present disclosure is in part based on the discovery that one or more microbial strains or compositions comprising one or more microbial strains are particularly suitable as therapeutic agents for insulin-related diseases, disorders, or afflictions.

[0095] Microbial agents and / or components

[0096] The present disclosure provides systems and methods for assessing, characterizing, and identifying one or more microbial strains of a microbiome. For example, the present disclosure provides systems and methods for assessing, characterizing, and identifying one or more microbial strains of a microbiome having one or more capabilities. Such systems and methods can be used to assess, characterize, and identify one or more microbial strains that affect the health of humans, livestock, and / or pets. In some embodiments, one or more microbial strains affect the health of humans, livestock, and / or pets by modulating their respective metabolomes, cell viability, ATP levels, one or more other parameters or characteristics (e.g., one or more parameters or characteristics of an organ of a subject), or combinations thereof, to prevent, treat a disease, disorder, or affliction, or reduce the risk of contracting a disease, disorder, or affliction. For example, the techniques described herein can result in modulating the metabolome of a subject, improving cell viability, increasing ATP levels, modulating one or more other parameters or characteristics (e.g., the level or activity or form of nucleic acids or proteins, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.), or combinations thereof, resulting in a reduction in the production of toxic components and / or components that imply or serve as markers of cell damage (e.g., hepatocyte damage, pancreatic cell damage, neuronal cell damage (e.g., increased blood levels of neurofilament light protein (NF-L)) in a subject (e.g., in the blood of a subject)).

[0097] The present disclosure also provides systems and methods for manufacturing a pharmaceutical composition, the method comprising assessing, characterizing, and identifying one or more microbial strains of a microbiome.

[0098] In some embodiments, one or more microbial strains of a microbiome from a snake, lizard, fish, or bird are assessed, characterized, and identified. In some embodiments, one or more microbial strains of a mammalian microbiome are assessed, characterized, and identified. The mammalian microbiome can be a canine, feline, equine, bovine, ovine, caprine, or porcine microbiome. In some embodiments, the microbiome used in the systems or methods described herein can prevent or treat a disease or affliction.

[0099] The microbiome can be isolated from any system or tissue of an organism that supports the growth of microorganisms. For example, the microbiome can be a skin microbiome, an oral microbiome, a nasal microbiome, a gastrointestinal microbiome, a brain microbiome, a lung microbiome, or a urogenital microbiome. A list of exemplary microbial strains found in the gastrointestinal microbiome is included in Table 1 below. Those skilled in the art will understand that microbiome samples can be obtained by various means known in the art. For example, swabs or tissue scrapings can be used to obtain skin, oral, nasal, lung, or urogenital microbiome samples. In some embodiments, the gastrointestinal microbiome can be sampled from feces. Skin microbiome, oral microbiome, nasal microbiome, gastrointestinal microbiome, brain microbiome, lung microbiome, or urogenital microbiome samples can be obtained by biopsy.

[0100] In some embodiments, the microbiome is that of a healthy individual or an individual who does not have or is not at risk of developing a particular disease or disorder. In some embodiments, the microbiome is that of an individual who has a particular disease, disorder, or affliction or is at risk of developing a particular disease, disorder, or affliction. In some embodiments, the microbiome is that of an individual known to have a particular disease, disorder, or affliction. In some embodiments, the human microbiome is that of a person who has an unknown risk of one or more diseases, disorders, or afflictions.

[0101] In some embodiments, the microbiome is a reference microbiome. The reference microbiome can be that of a healthy individual or an individual who does not have a particular disease, disorder, or affliction or is not at risk of developing a particular disease, disorder, or affliction. In some cases, the reference microbiome may be from the same individual as the microbiome to be evaluated or characterized, but obtained at a different time. In some cases, the reference microbiome may be from the same individual as the microbiome to be evaluated or characterized, but obtained from a different system or tissue.

[0102] In some embodiments, individual microbial strains or combinations of microbial strains can be evaluated, characterized, or identified in relative amounts different from such one or more strains found in the microbiome. For example, in vitro methods as described herein (e.g., mammalian cells) or in vivo methods using mammals (e.g., mice, humans, etc.) can be used to evaluate, characterize, or identify the regulatory effect of a cell or organism in response to a single strain. For example, in some embodiments, in vitro methods as described herein (e.g., mammalian cells) or in vivo methods using mammals (e.g., mice, humans, etc.) can be used to evaluate, characterize, or identify the effect of modulating a cell or organism to treat, prevent a disease, disorder, or affliction or reduce the risk of a disease, disorder, or affliction (e.g., an insulin-related disease, disorder, or affliction as described herein). In some embodiments, in vitro methods as described herein (e.g., mammalian cells) or in vivo methods using mammals (e.g., mice, humans, etc.) can be used to evaluate, characterize, or identify the effect of modulating a cell or organism to treat, prevent a disease, disorder, or affliction (e.g., an insulin-related disease, disorder, or affliction as described herein) or reduce its risk, for example, by modulating one or more metabolites of the cell or organism, one or more characteristics or parameters of the cell or organism (e.g., cell viability, levels or activity of nucleic acids or proteins or their forms, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) or a combination thereof. As another example, the methods described herein can be used together to evaluate, characterize, or identify the effect of modulating a cell or organism (e.g., levels of one or more metabolites) in response to two microbial strains to treat, prevent a disease, disorder, or affliction as described herein or reduce its risk.

[0103] The methods described herein can also be used to evaluate, characterize, or identify extracts, components, or compounds of microbial strains. In some cases, extracts, components, or compounds of microbial strains that have been determined to be useful for treating, preventing a disease, disorder, or affliction in an organism (e.g., a mammalian) as described herein or reducing its risk can be evaluated, characterized, or identified. Evaluating, characterizing, or identifying extracts, components, or compounds of microbial strains that treat, prevent a disease, disorder, or affliction in an organism (e.g., a mammalian) or reduce its risk can provide additional information about potential biomarkers, targets, or protective agents in the microbiome.

[0104] A variety of techniques are known in the art for preparing extracts of microbial strains, and / or for isolating extracts, components, or compounds therefrom, or for processing (e.g., separating and / or purifying one or more components or compounds therefrom). By way of example only, such techniques can include one or more of, for example, organic extraction, vacuum concentration, chromatography, etc.

[0105] Evaluating biological effects

[0106] The present disclosure provides the recognition that the compositions as described herein (e.g., microbiome compositions) can be used to treat, prevent a disease, disorder or condition and / or reduce the risk thereof in an organism (e.g., a mammal (e.g., a human)) by contacting the composition (e.g., feeding, administering the composition to the organism). In some embodiments, the organism may have a disease, disorder or condition (e.g., a mammalian disease, disorder or condition) or be at risk of developing a disease, disorder or condition. To determine whether one or more compositions treat, prevent a disease, disorder or condition (e.g., an insulin-related disease, disorder or condition) or reduce the risk thereof, the levels of one or more metabolites in a sample that has been contacted with one or more compositions can be observed, measured or evaluated. For example, the levels of one or more metabolites in the sample can be observed, measured or evaluated at different times (e.g., before administering the composition, after administering the composition, during administering the composition, etc.). To determine whether one or more compositions treat, prevent a disease, disorder or condition (e.g., an insulin-related disease, disorder or condition) or reduce the risk thereof, one or more characteristics or parameters in a sample that has been contacted with one or more compositions can be observed, measured or evaluated. For example, one or more characteristics or parameters in the sample can be observed, measured or evaluated at different times (e.g., before administering the composition, after administering the composition, during administering the composition, etc.).

[0107] In some embodiments, the methods described herein utilize a first sample and a second sample. In some embodiments, the first sample is a reference sample. In some embodiments, the reference sample can be a sample obtained from a subject that has been contacted (e.g., administered or fed) with a composition (such as a CT10 composition, a CT10m composition, a CT10x composition, a CT6 composition, or a CT6m composition). In some embodiments, the reference sample can be a sample obtained from a subject at a first time point that has been contacted (e.g., administered or fed) with a composition (such as a CT10 composition, a CT10x composition, a CT10m composition, a CT6 composition, or a CT6m composition). In some embodiments, the reference sample can be a sample obtained from a subject prior to being contacted (e.g., administered or fed) with a composition (such as a CT10 composition, a CT10x composition, a CT10m composition, a CT6 composition, or a CT6m composition). In some embodiments, the reference sample can be a sample obtained from a healthy individual. In some embodiments, the reference sample can be a sample obtained from an individual having a disease, disorder, or condition (such as an insulin-related disease, disorder, or condition) or at risk of having a disease, disorder, or condition. In some embodiments, the reference sample is a control sample. In some embodiments, the reference sample is a negative control sample. In some embodiments, the reference sample is a positive control sample. In some embodiments, the reference sample can be a historical reference (e.g., values across control samples). In some embodiments, the reference sample can be from a printed publication (such as a textbook, a journal, etc.).

[0108] In some embodiments, the second sample can be a test sample. In some embodiments, the test sample can be a sample obtained from a subject that has been contacted (e.g., administered or fed) with a composition (such as a CT10 composition, a CT10x composition, a CT10m composition, a CT6 composition, or a CT6m composition). In some cases, the subject (such as a patient or a group) may have a disease, disorder, or condition (such as an insulin-related disease, disorder, or condition) or be at risk of a disease, disorder, or condition. In some cases, the subject (such as a patient or a group) may have an unknown risk of developing one or more diseases, disorders, or conditions as described herein. In some embodiments, the test sample can be a sample obtained from a subject at a second time point that has been contacted (e.g., administered or fed) with a composition (such as a CT10 composition, a CT10x composition, a CT10m composition, a CT6 composition, or a CT6m composition).

[0109] In some embodiments, the methods described herein include comparing one or more metabolite levels (e.g., metabolome) obtained from a test sample, or one or more parameters or characteristics (e.g., cell viability, levels or activities or forms of nucleic acids or proteins, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) with one or more metabolite levels (e.g., metabolome) obtained from a reference sample, or one or more parameters or characteristics (e.g., cell viability, levels or activities or forms of nucleic acids or proteins, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.). In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, the compositions described herein can be evaluated, characterized, or identified as useful for treating, preventing a disease, disorder, or condition as described herein (e.g., an insulin-related disease, disorder, or condition) or reducing the risk of developing the disease, disorder, or condition. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that the compositions disclosed herein increase the severity or incidence of a disease, disorder, or condition phenotype. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that the compositions disclosed herein reduce the severity or incidence of a disease, disorder, or condition phenotype. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that the compositions disclosed herein have no effect on the severity or incidence of a disease, disorder, or condition phenotype. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that the compositions disclosed herein prevent a disease, disorder, or condition phenotype.

[0110] The present disclosure also provides the recognition that the compositions and methods provided herein can be used to monitor the progression of a disease, disorder, or affliction (e.g., an insulin-related disease, disorder, or affliction) in an individual. For example, if a metabolite level, parameter, or characteristic (e.g., cell viability, level or activity of a nucleic acid or protein or a form thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride level, blood cholesterol level, oxidative stress, etc.) that is determined to increase the severity of a disease, disorder, or affliction decreases in relative amount, it can indicate that the disease, disorder, or affliction is being attenuated, e.g., through treatment or an immune response.

[0111] The present disclosure also provides the recognition that the compositions and methods provided herein can be used to customize treatment (e.g., therapies, nutritional products, and / or probiotics) for an individual patient. In some embodiments, the compositions and methods provided herein can provide a "personalized" therapy. In some cases, metabolite levels, characteristics, or parameters (e.g., cell viability, level or activity of a nucleic acid or protein or a form thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride level, blood cholesterol level, oxidative stress, etc.) in an individual can be evaluated, characterized, or identified to determine whether they have a disease, disorder, or affliction. Based on the results, the individual can be treated with one or more compositions to adjust the metabolite levels (i.e., their metabolome), characteristics, or parameters. In some cases, this can affect the disease, disorder, or affliction that the individual has or is at risk of developing. For example, if it is determined that an individual has a relatively low amount of one or more metabolite levels that have been determined to reduce the severity of a disease, disorder, or affliction, administering one or more compositions (or extracts, components, or compounds thereof) that have been determined to reduce the severity of the individual's disease, disorder, or affliction can reduce the severity of the individual's disease or affliction.

[0112] The present disclosure provides the insight that the compositions and methods provided herein can be used recursively to treat, prevent, or ameliorate a disease, disorder, or condition. In some embodiments, for example, after determining the effect of one or more compositions on the metabolite levels of a subject, or after determining the effect of one or more compositions on a characteristic or parameter of a subject (such as cell viability, the level or activity of nucleic acids or proteins or their forms, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.), one or more compositions disclosed herein can be administered (such as by feeding, injection, etc.) to the subject. In some embodiments, the composition can be administered once. In some embodiments, the composition can be administered more than once. In some embodiments, the composition can be administered daily, weekly, bi-weekly, monthly, bi-monthly, etc. In each of these cases, the change in the level of one or more metabolites or the change in a characteristic or parameter can be monitored. In some embodiments, the change in the level of one or more metabolites (such as the metabolome) or the change in a characteristic or parameter can be monitored before administering the composition. In some embodiments, the change in the level of one or more metabolites (such as the metabolome) or the change in a characteristic or parameter can be monitored after administering the composition.

[0113] Pharmaceutical compositions

[0114] Compositions are provided herein that comprise a single microbial strain or a combination of microbial strains, their metabolites, their extracts, or their components. In some embodiments, the composition comprises a single microbial strain or a combination of microbial strains, their metabolites, their extracts, and / or their components from the mammalian microbiome that have been evaluated, identified, characterized, or assayed using the methods described herein. In some embodiments, the compositions provided herein comprise one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more microbial strains, their extracts, their metabolites, and / or their components from the mammalian microbiome that have been evaluated, identified, characterized, or assayed using the methods described herein.

[0115] Compositions are also provided herein that comprise one or more components or metabolites. In some embodiments, the components or metabolites in the compositions herein are from non-microbial strain sources, for example, synthetically produced. In some embodiments, the components or metabolites in the composition may have been identified from a microbial strain but are independent of and not produced by the microbial strain, for example, they can be synthetically produced.

[0116] In some embodiments, the compositions provided herein comprise two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more microbial strains listed in Table 1 below.

[0117] Table 1: Exemplary microbial strains found in the human gut microbiome

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133] In some embodiments, the compositions provided herein comprise Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or combinations thereof. In some embodiments, the composition comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all of the following: Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, and Acidaminococcus sp. In some embodiments, for example, the composition comprises all of Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, and Acidaminococcus sp., and can be referred to by different names, including but not limited to CT10 composition, CT10 mixture, and the like.

[0134] In some embodiments, the compositions provided herein comprise Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or combinations thereof. In some embodiments, the composition comprises at least two, at least three, at least four, at least five, or all of the following: Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., and Bifidobacterium sp. In some embodiments, for example, the composition comprises all of Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., and Bifidobacterium sp., and can be referred to by different names, including but not limited to CT6 composition, CT6 mixture, and the like. In some embodiments, the compositions provided herein comprise Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or combinations thereof. In some embodiments, the composition comprises at least two, at least three, at least four, at least five, or all of the following: Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, and Bifidobacterium breve. In some embodiments, for example, the composition comprises all of Acetobacter hansenii, Geosporobacter ethanolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, and Bifidobacterium breve, and can be referred to by different names, including but not limited to CT6 composition, CT6 mixture, and the like.

[0135] Exemplary microbiome compositions as described herein (e.g., CT10, CT10m, CT10x, CT6, CT6m) and details of their compositions are listed in Tables 2-6 below.

[0136] Table 2: CT10 composition

[0137] # Microbiome therapy (MBT) Concentration (CFU / ml) 1 Bifidobacterium breve <![CDATA[1x 10 9 > 2 Gluconacetobacter hansenii <![CDATA[1x 10 9 > 3 Solibacillus glycolicus <![CDATA[1x 10 9 > 4 Veillonella atypica <![CDATA[1x 10 9 > 5 Lactobacillus plantarum <![CDATA[1x 10 9 > 6 Coprococcus catus <![CDATA[1x 10 9 <!-- 35 -->]]> 7 Clostridium butyricum <![CDATA[1x 10 9 > 8 Paenibacillus barengoltzii <![CDATA[1x 10 9 > 9 Bacillus subtilis <![CDATA[1x 10 9 > 10 Aminococcus sp. <![CDATA[1x 10 9 >

[0138] Table 3: CT10m composition

[0139]

[0140]

[0141] Table 4: CT10x composition

[0142] # Microbiome therapy (MBT) Concentration (CFU / ml) 1 Bifidobacterium breve <![CDATA[1x 10 9 > 2 Gluconacetobacter hansenii <![CDATA[3x 10 9 > 3 Solibacillus glycolicus <![CDATA[1x 10 9 > 4 Veillonella atypica <![CDATA[3x 10 9 > 5 Lactobacillus plantarum <![CDATA[3x 10 9 > 6 Coprococcus catus <![CDATA[1x 10 9 > 7 Clostridium butyricum <![CDATA[1x 10 9 > 8 Paenibacillus barengoltzii <![CDATA[1x 10 9 > 9 Bacillus subtilis <![CDATA[1x 10 9 > 10 Aminococcus sp. <![CDATA[1x 10 9 >

[0143] Table 5: CT6 composition

[0144] # MBT Concentration (CFU / ml) 1 Bifidobacterium breve <![CDATA[1x 10 9 > 2 Gluconacetobacter hansenii <![CDATA[1x 10 9 > 3 Solibacillus glycolicus <![CDATA[1x 10 9 > 4 Veillonella atypica <![CDATA[1x 10 9 > 5 Lactobacillus plantarum <![CDATA[1x 10 9 > 6 Coprococcus catus <![CDATA[1x 10 9 >

[0145] Table 6: CT6m composition

[0146] # MBT Concentration (CFU / ml) 1 Bifidobacterium breve <![CDATA[1x 10 9 > 2 Gluconacetobacter hansenii <![CDATA[3x 10 9 > 3 Solibacillus glycolicus <![CDATA[1x 10 9 > 4 Veillonella atypica <![CDATA[3x 10 9 > 5 Lactobacillus plantarum <![CDATA[1x 10 9 > 6 Coprococcus catus <![CDATA[1x 10 9 >

[0147] In some embodiments, the compositions provided herein comprise one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more metabolites. Metabolites that can be evaluated, identified, characterized, or determined and / or included in the compositions disclosed herein include, for example, those listed in the appendices filed herewith (e.g., Appendices 1-1, 1-2, 1-3, 2, 3, or 4).

[0148] In some embodiments, the metabolite can be butyroyl carnitine, theobromine, 4-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.

[0149] In some embodiments, the metabolite can be 4-hydroxyphenylpyruvic acid, ectoine, gramine, N-acetyl-L-phenylalanine, Nε-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyryl carnitine, b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenylacetylglycine, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013C6H6O4S, 3-indolylsulfuric acid, nicotinamide, N-formylglycine, ureidoethanolate, N-methylproline, glucaric acid, butyryl carnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenylacetylglycine, diethanolamine, phosphocholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thioproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0XA0027 or any combination thereof.

[0150] In some embodiments, a single microbial strain or a combination of microbial strains from the mammalian microbiome has been killed (e.g., heat killed). Alternatively, in some embodiments, a single microbial strain or a combination of microbial strains from the mammalian microbiome can include viable or live cells.

[0151] In some embodiments, one or more microbial strains comprise a viable or live single microbial strain or a combination of microbial strains, such as from the mammalian microbiome.

[0152] In some embodiments, one or more microbial strains comprise a viable or live single microbial strain or a combination of microbial strains, such as from the mammalian microbiome, comprising one or more cell cultures and / or supernatants or precipitates thereof, and / or powders formed therefrom, and / or formulated using one or more cell cultures and / or supernatants or precipitates thereof, as described herein.

[0153] In some embodiments, the compositions used in accordance with the present disclosure are pharmaceutical compositions, for example for administration (e.g., topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal (e.g., rectal intubation), ocular, intravitreal or suprachoroidal administration) to a mammal (e.g., a human). The pharmaceutical compositions generally include an active agent (e.g., a single microbial strain or combination of microbial strains from the mammalian microbiome, extracts thereof, and / or components thereof) and a pharmaceutically acceptable carrier. Certain exemplary pharmaceutically acceptable carriers include, for example, saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption delaying agents compatible with drug administration, etc.

[0154] In some embodiments, the pharmaceutical compositions used in accordance with the present disclosure may comprise one or more additional active compounds and / or may be administered in combination with one or more additional active compounds; in certain embodiments, such additional active agents may include ginger, curcumin, probiotics (e.g., probiotic strains of one or more of the following genera: Lactobacillus, Bifidobacterium, Saccharomyces, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, and / or Escherichia coli (see Fijan, Int J Environ Res Public Health. May 2014; 11(5):4745–4767, which is incorporated herein by reference in its entirety)); prebiotics (indigestible food ingredients that help support the growth of probiotics, such as fructans like fructooligosaccharides (FOS) and inulin, galactans like galactooligosaccharides (GOS), dietary fibers like resistant starch, pectin, β-glucan, and xylooligosaccharides (Hutkins et al., Curr Opin Biotechnol. February 2016; 37:1–7, which is incorporated herein by reference in its entirety)) and combinations thereof.

[0155] In some embodiments, the prebiotics include fructooligosaccharides, inulin, isomaltooligosaccharides, lactitol, lactooligosaccharides, lactulose, soy oligosaccharides, trans-galactooligosaccharides, xylooligosaccharides, seaweed, or combinations thereof. In some embodiments, the prebiotics include seaweed. In some embodiments, the prebiotics include extracts of pears, berries, and walnuts.

[0156] In some embodiments, the probiotic composition can be formulated for oral administration. In some embodiments, the probiotic composition can be a food, beverage, feed composition, or nutritional supplement. In some embodiments, the ellagitannin composition, enzyme composition, or both can be a liquid, syrup, tablet, lozenge, gummy, capsule, powder, gel, or film. In some embodiments, the probiotic composition is an enteric-coated preparation.

[0157] In some embodiments, the probiotic contains prebiotics. In some embodiments, the prebiotics include fructooligosaccharides, inulin, isomaltooligosaccharides, lactitol, lactosucrose, lactulose, soy oligosaccharides, trans-galactooligosaccharides, xylooligosaccharides, seaweed, pear fruit extract, berry extract, and walnut extract, or a combination thereof.

[0158] The pharmaceutical composition is generally formulated to be compatible with its intended route of administration. Examples of routes of administration include topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal (e.g., rectal intubation), ocular, intravitreal, or suprachoroidal administration. Methods for formulating suitable pharmaceutical compositions are known in the art, see, for example, Remington: The Science and Practice of Pharmacy, 21st Edition, 2005; and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY), which are incorporated herein by reference in their entirety. Oral compositions generally contain an inert diluent or an edible carrier (e.g., a pharmaceutically acceptable diluent, a pharmaceutically acceptable carrier). By way of example only, in some embodiments, the oral preparation can be or include syrups, liquids, tablets, lozenges, gummies, capsules (e.g., gelatin capsules), powders, gels, films, etc. Similarly, ophthalmic compositions (e.g., for ocular, intravitreal, or suprachoroidal administration) can contain an inert diluent or a carrier (e.g., a pharmaceutically acceptable diluent, a pharmaceutically acceptable carrier), various additives such as viscosity enhancers, penetration enhancers, cyclodextrins, etc. Examples of viscosity enhancers include hydroxyethyl methylcellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, and polyols. Examples of penetration enhancers include chelating agents, preservatives, surfactants, bile salts, benzalkonium chloride, polyethylene glycol ethers (lauryl, stearyl, and oleyl), sodium salts of ethylenediaminetetraacetic acid, sodium taurocholate, saponins, and cremophor EL, etc. For example, in some embodiments, the ophthalmic preparation can be or include suspensions, emulsions (e.g., oil-in-water or water-in-oil), nanocarriers (e.g., nanoparticles, nanosuspensions, liposomes, nanomicelles, dendrimers, etc.), ointments, gels, eye drops, etc. Cerebral compositions (e.g., for intracerebral or intrathecal administration) can contain an inert diluent or a carrier and / or additives. In some embodiments, the cerebral composition does not contain preservatives. In some embodiments, the cerebral composition is sterile.

[0159] In some embodiments, a pharmaceutically compatible binder and / or adjuvant material may be included as part of the pharmaceutical composition. In some specific embodiments, the pharmaceutical composition may contain any one or more of, for example, the following inactive ingredients or compounds having similar properties: binders such as microcrystalline cellulose, tragacanth or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel or corn starch; lubricants such as magnesium stearate or Sterote; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate or orange flavoring. In some embodiments, the composition may be taken as is or sprayed onto or mixed into food or a liquid (such as water). In some embodiments, the composition that may be administered to a mammal as described herein may be or comprise an ingestible article (such as a food or beverage) that contains (such as is supplemented with) a single microbial strain or a combination of microbial strains, an extract thereof, and / or a component thereof from the mammalian microbiome.

[0160] In some embodiments, the food may be or comprise one or more of a bar, confectionery, baked good, cereal, savory snack, pasta, chocolate and other solid foods, as well as liquid or semi-solid foods (including yogurt, soup and stew), and beverages such as smoothies, milkshakes, fruit juices and other carbonated or non-carbonated drinks. In some embodiments, the food is prepared by the subject by mixing a single microbial strain or a combination of microbial strains, an extract thereof, and / or a component thereof from the mammalian microbiome.

[0161] The composition may be included in a kit, container, package or dispenser, together with instructions for administration or for use in the methods described herein.

[0162] Those skilled in the art reading this disclosure will understand that, in some embodiments, the composition as described herein (such as a pharmaceutical composition) may be or comprise one or more cells, tissues or organisms (such as plant or microbial cells, tissues or organisms) that produce (such as have produced and / or are producing) the relevant compound.

[0163] Those skilled in the art will understand that, in some embodiments, the techniques for preparing a composition and / or a formulation and / or for preparing (and in particular for preparing a pharmaceutical composition) may include one or more steps of evaluating or characterizing the compound, formulation or composition, for example as part of quality control. In some embodiments, if the material being assayed does not meet the predetermined specifications of the relevant evaluation, it is discarded. In some embodiments, if the material so assayed does meet the predetermined specifications, it proceeds to be processed as described herein.

[0164] In some embodiments, the pharmaceutical compositions provided herein can promote the colonization of individual microbial strains or combinations of microbial strains from the mammalian microbiome, particularly microbial strains that have been identified, characterized, or evaluated as reducing the severity or incidence of a mammalian disease, disorder, or condition in a mammal having or at risk of having a mammalian disease, disorder, or condition. In some embodiments, the pharmaceutical compositions provided herein can attenuate the colonization of individual microbial strains or combinations of microbial strains from the mammalian microbiome, particularly microbial strains that have been identified, characterized, or evaluated as increasing the severity or incidence of a mammalian disease, disorder, or condition (e.g., insulin-related disease, disorder, or condition) in a mammal having or at risk of having a mammalian disease, disorder, or condition. In some embodiments, the pharmaceutical compositions provided herein can promote the colonization of individual microbial strains or combinations of microbial strains from the mammalian microbiome, particularly microbial strains that have been identified, characterized, or evaluated as having no effect on the severity or incidence of a mammalian disease, disorder, or condition in a mammal having or at risk of having a mammalian disease, disorder, or condition, but have been identified, characterized, or evaluated as being able to outcompete one or more microbial strains that have been identified, characterized, or evaluated as increasing the severity or incidence of a mammalian disease, disorder, or condition.

[0165] In some embodiments, each of the one or more microbial strains in the composition comprises 10 1 colony forming units (CFU) to 10 20 CFU. In some embodiments, each of the one or more microbial strains in the composition comprises 10 1 colony forming units (CFU) to 10 15 CFU. In some embodiments, each of the one or more microbial strains in the composition comprises 10 6 CFU to 10 15 CFU. In some embodiments, each of the one or more microbial strains in the composition comprises about 10 1 CFU to 10 15 CFU, or about 10 2 CFU to 10 14 CFU, or about 10 3 CFU to 10 13 CFU, or about 10 4 CFU to 10 13 CFU, or about 10 5 CFU to 10 12 CFU, or about 10 6 CFU to 10 11CFU, or about 10 7 CFU to 10 10 CFU, or about 10 8 CFU to 10 9 CFU, or about 10 5 CFU to 10 10 CFU, or about 10 8 CFU to 10 12 CFU. In some embodiments, each of one or more microbial strains in the composition comprises at least about 10 1 、5x 10 1 、10 2 、5x 10 2 、10 3 、5x 10 3 、10 4 、5x 10 4 、10 5 、5x 10 5 、10 6 、5x 10 6 、10 7 、5x 10 7 、10 8 、5x 10 8 、10 9 、5x 10 9 、10 10 、5x 10 10 、10 11 、5x 10 11 、10 12 or more CFU. In some embodiments, each of one or more microbial strains in the composition comprises at most about 10 15 、5x 10 14 、10 14 、5x 10 13 、10 13 、5x 10 12 、10 12 、5x 10 11 、10 11 、5x10 10 、10 10 、5x 10 9 、10 9 、5x 10 8 、10 8 or fewer CFU. In some embodiments, each of one or more microbial strains in the composition comprises the same number of CFU. In some embodiments, some of one or more microbial strains in the composition comprise different numbers of CFU.

[0166] In some embodiments, the composition comprises a total of 10 1 CFU to 10 20 CFU. In some embodiments, the composition comprises a total of 10 6 CFU to 10 15 CFU. In some embodiments, the composition may comprise from about 10 1 CFU to 10 20 CFU, or from about 10 5 CFU to 10 15 CFU, or from about 10 5 CFU to 10 12 CFU, about 10 5 CFU to 10 10 CFU, or from about 10 8 CFU to 10 12 CFU of one or more microbial strains. In some embodiments, the composition may comprise from about 10 1 CFU to 10 15 CFU, or from about 10 2 CFU to 10 14 CFU, or from about 10 3 CFU to 10 13 CFU, or from about 10 4 CFU to 10 13 CFU, or from about 10 5 CFU to 10 12 CFU, or from about 10 6 CFU to 10 11 CFU, or from about 10 7 CFU to 10 10 CFU, or from about 10 8 CFU to 10 9 CFU, or from about 10 5 CFU to 10 10 CFU, or from about 10 8 CFU to 10 12 CFU of one or more microbial strains. In some embodiments, the composition may comprise at least 10 1 , 5x 10 1 , 10 2 , 5x 10 2 , 10 3 , 5x 10 3 , 10 4 , 5x 10 4 , 10 5 , 5x 10 5 , 10 6 , 5x 10 6 , 107 、 5x 10 7 、 10 8 、 5x 10 8 、 10 9 、 5x10 9 、 10 10 、 5x 10 10 、 10 11 、 5x 10 11 、 10 12 or more CFU of one or more microbial strains. In some embodiments, the composition can comprise up to 10 15 、 5x 10 14 、 10 14 、 5x 10 13 、 10 13 、 5x 10 12 、 10 12 、 5x 10 11 、 10 11 、 5x 10 10 、 10 10 、 5x 10 9 、 10 9 、 5x 10 8 、 10 8 or fewer CFU of one or more microbial strains.

[0167] In some embodiments, the pharmaceutical composition is customized for a particular mammal (e.g., a particular human, e.g., a patient) based on the microbiome of the particular mammal. In some embodiments, the pharmaceutical composition is specific to the microbiome of an individual mammal (e.g., a human). In some embodiments, the pharmaceutical composition is specific to the microbiome of a population of mammals. The population of mammals can include, but is not limited to: families, mammals in the same regional location (e.g., community, city, state, or country), mammals suffering from the same disease or disorder, mammals of a particular age or age range, mammals consuming a particular diet (e.g., food, food source, or caloric intake).

[0168] Treatment methods

[0169] The present disclosure recognizes that the compositions described herein can be used to treat a subject. The methods provided by the present disclosure include methods for treating certain diseases, conditions, and disorders. In some embodiments, the relevant diseases, conditions, and disorders can be or include insulin-related diseases, conditions, or disorders. In some embodiments, the insulin-related diseases, conditions, or disorders can be ALS, AD, PD, or HD.

[0170] Generally speaking, the treatment methods provided by the present disclosure include administering a therapeutically effective amount of the compositions as described herein to a subject in need of such treatment or determined to be in need of such treatment, either alone or in combination with other compositions and / or treatments.

[0171] In some embodiments, the treatment methods provided herein are prophylactic or preventive, for example, they can be administered to a subject before presenting symptoms and / or being exposed to specific expected triggers related to the insulin-related diseases, disorders or afflictions described herein. In some embodiments, the treatment methods provided herein are therapeutic, for example, they can be administered to a subject after significant symptoms related to insulin-related diseases, disorders or afflictions appear.

[0172] In some embodiments, the provided treatment methods are administered to a subject that is a mammal, such as a mammal suffering from the diseases, disorders or afflictions as described herein; in some embodiments, the subject is a human or non-human veterinary subject, such as an ape, a cat, a dog, a monkey or a pig.

[0173] In many embodiments, the treatment involves improving at least one symptom of the diseases, disorders or afflictions related to insulin-related diseases, disorders or afflictions. In some embodiments, the treatment method can be prophylactic.

[0174] In some embodiments, the method may include administering a therapeutically effective amount of the compositions disclosed herein before, during (e.g., simultaneously) or after administering a treatment expected to be related to insulin-related diseases, disorders or afflictions.

[0175] In some embodiments, a subject receiving the treatment as described herein may be receiving and / or may have received other treatments (e.g., drug treatment / therapy, surgery, etc.), for example, treatments that may be intended to treat one or more symptoms or features of the diseases, disorders or afflictions as described herein (e.g., insulin-related diseases, disorders or afflictions), such that the provided compositions are administered in combination with such other therapies (i.e., treatments) to treat the related diseases, disorders or afflictions.

[0176] In some embodiments, the compositions described herein can be administered in the form of one or more pharmaceutically acceptable carriers. Suitable carriers have been previously described and vary with the desired form of the composition and the mode of administration. For example, pharmaceutically acceptable carriers can include diluents or excipients, such as fillers, binders, wetting agents, disintegrants, surfactants, glidants and lubricants. Generally, the carrier can be a solid (including powder), a liquid or any combination thereof. Each carrier is preferably "acceptable" in the sense that it is compatible with the other components in the composition and is harmless to the subject. The carrier can be biocompatible and inert (e.g., it allows the composition to maintain the viability of the biological material until delivered to the appropriate site).

[0177] Tablets, pills, capsules, troches, and the like may contain any of the following ingredients or compounds of a similar nature: binders such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch or lactose; disintegrating agents such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or Sterotes; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, orange flavoring, or other suitable flavoring agents. These are for illustrative purposes only and are not intended to be limiting.

[0178] Oral compositions may contain an inert diluent or an edible carrier. For purposes of oral therapeutic administration, the active compounds may be combined with excipients and used in the form of tablets, lozenges, troches, or capsules (e.g., gelatin capsules). Oral compositions may also be prepared by combining the compositions of the present disclosure with food. In some embodiments, microorganisms (e.g., one or more microbial strains) may be formulated in food. Some non-limiting examples of foods for use with the methods and compositions described herein include: ice pops, cheese, cream, chocolate, milk, meat, beverages, pickled vegetables, kefir, miso, sauerkraut, and the like. In other embodiments, the food may be: fruit juices, soft drinks, tea beverages, beverage products, jelly drinks, and functional drinks; alcoholic beverages such as beer; carbohydrate-containing foods such as rice food products, noodles, bread, and pasta; paste products such as fish paste, ham, sausage, and seafood paste products; retort pouch products such as curry, foods made with thick starch sauce, and Chinese soups; soups; dairy products such as milk, dairy beverages, ice cream, and yogurt; fermented products such as fermented soybean paste, fermented beverages, and pickled vegetables; soy products; various confectionery products including cookies, crackers, etc., candies, chewing gum, gummy candies, frozen desserts (including jelly, crème caramel, and frozen desserts); ready-to-eat foods such as instant soups and instant soybean soups; and the like. Food preparations that do not require cooking after mixing with the microbial strain are preferred to avoid killing any microorganisms. In one embodiment, the food for administration is cold, such as chilled flavored water. In certain embodiments, the food is not a potentially allergenic food (e.g., is not soy, wheat, peanuts, nuts, dairy, eggs, shellfish, or fish). Pharmaceutically compatible binders and / or adjuvant substances may be included as part of the composition.

[0179] Ophthalmic preparations (e.g., for ophthalmic, intravitreal, or suprachoroidal administration) may contain an inert diluent or carrier. For the purpose of therapeutic ophthalmic administration, the active compound may be mixed with excipients and used in the form of suspensions, emulsions (e.g., water-in-oil or oil-in-water), nanocarriers (e.g., nanoparticles, nanosuspensions, liposomes, nanomicelles, dendrimers, etc.), ointments, gels, eye drops, etc. In some embodiments, the administration of such preparations is topical (e.g., eye drops). In some embodiments, the administration of such preparations is by injection (e.g., intravitreal, suprachoroidal, etc.).

[0180] Brain preparations (e.g., for intracerebral or intrathecal administration) may contain an inert diluent or carrier. For the purpose of therapeutic brain administration, the active compound may be mixed with excipients and used in the form of suspensions, emulsions (e.g., water-in-oil or oil-in-water), nanocarriers (e.g., nanoparticles, nanosuspensions, liposomes, nanomicelles, dendrimers, etc.), ointments, gels, etc. In some embodiments, the administration of such preparations is topical (e.g., ointment). In some embodiments, the administration of such preparations is by injection (e.g., intracerebral, intrathecal, etc.).

[0181] In some such embodiments, the compositions described herein are administered to a subject according to a dosing regimen that achieves a population of the subject's microbiome and the administered cells. In some embodiments, the composition is administered to the subject as a single dose. In some embodiments, the composition is administered to the subject in multiple doses. In some embodiments, a dose of the composition is administered to the subject twice daily, once daily, once weekly, or once monthly.

[0182] In some embodiments, each of one or more microbial strains in a dose contains 10 1 to 10 15 colony forming units (CFU). In some embodiments, each of one or more microbial strains in a dose contains 10 6 to 10 15 CFU. In some embodiments, each of one or more microbial strains in a dose contains the same number of CFU. In some embodiments, some of one or more microbial strains in a dose contain different numbers of CFU.

[0183] In some embodiments, a dose of one or more microbial strains contains a total of 10 6 to 10 15 CFU. In some embodiments, a dose of one or more microbial strains contains a total of 10 7 to 10 15CFU. In some embodiments, the dose of one or more microbial strains comprises 50 - 200 billion CFU. In some embodiments, the dose of one or more microbial strains comprises 50 - 50 billion CFU. In some embodiments, the dose of one or more microbial strains comprises 50 - 20 billion CFU. In some embodiments, the dose of one or more microbial strains comprises 50 - 100 billion CFU. In some embodiments, the dose of one or more microbial strains comprises 100 - 200 billion CFU.

[0184] In some embodiments, efficacy can be evaluated by measuring the degree of oxidative stress of cells in a biological sample before and after administering the composition as described herein. The degree of oxidative stress of cells can be evaluated by, for example, measuring the expression of oxidative stress biomarkers (such as the level of reactive oxygen species (ROS) or the level of lipid, protein, and nucleic acid damage) or by determining the ratio of the oxidized form to the reduced form of one or more biomarkers. High levels of oxidative stress can be cytotoxic, and thus the degree of oxidative stress can be measured by evaluating the concentration of intracellular proteins present in the systemic circulation from inflamed or lysed cells (such as nerve cells).

[0185] Example

[0186] Example 1 : To evaluate the efficacy of the microbiome composition in affecting insulin signaling and its upstream molecule IDE in an AD mouse model

[0187] This example provides an evaluation of the efficacy of the microbiome composition (specifically, CT10, CT10x, and CT10m compositions) in affecting insulin signaling and its upstream molecule IDE related to Aβ clearance and neuroprotection in an in vivo mouse model of AD.

[0188] Background:The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter and memory functions, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). Several Aβ-degrading enzymes, including neprilysin (NEP), insulin-degrading enzyme (IDE), and endothelin-converting enzyme, reduce Aβ levels and prevent cognitive impairment in mouse models of AD (Miners et al., 2011). Insulin-degrading enzyme (IDE) is a thiol-metalloprotease that degrades multiple peptide hormones, including insulin, glucagon, atrial natriuretic peptide (ANP), and IGF-II (Duckworth et al., 1998). IDE has also been identified as the major protease involved in the degradation of Aβ peptides (Kurochkin and Goto, 1994; Vekrellis et al., 2000; Farris et al., 2003, 2004). Reduced levels of IDE protein and mRNA have been observed in the hippocampus and cortex of AD patients with the apolipoprotein E-e4 (ApoE4) allele (Cook et al., 2003; Zhao et al., 2004).

[0189] Mouse model: Nine-month-old Tg2576 mice were used for these experiments. This mouse model is one of the most popular transgenic mouse models, which overexpresses a mutant form of amyloid precursor protein (APP) (isoform 695) with the Swedish mutation (KM670 / 671NL), resulting in increased amyloid-β (Aβ) levels and ultimately the formation of amyloid plaques. Wild-type mice were used as controls in all experiments.

[0190] Study: The Tg2576 mice were divided into five groups of 15 animals each and were provided with either mock (DPBS) or the microbiome composition (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, followed by histological analysis.

[0191] Methods:To determine whether CT10, CT10x or CT10m treatment affects insulin signaling and its upstream molecules related to Aβ clearance and neuroprotection, the protein level of insulin-degrading enzyme (IDE) in brain samples of TG2576 (an AD mouse model) was examined by Western blotting. The major parts of the cortex, hippocampus and thalamus of brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using sodium dodecyl sulfate (SDS) sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom-made SDS-polyacrylamide Bis-tris gel (4%-12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) protein-free blocking buffer (catalog number 927-80001, LI-COR) or 5% non-fat milk powder solution and incubated overnight at 4 °C with the IDE-specific antibody (catalog number AB 9210, Millipore) diluted 1:1000 in T20 (TBS) protein-free antibody dilution buffer (catalog number 927-85001, LI-COR) under gentle shaking conditions. The next day, the membrane was washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in a 1:2000 dilution of horseradish peroxidase (HRP)-conjugated secondary anti-rabbit antibody (catalog number 7074; Cell signaling). After visualization with a G:Box Mini (Syngene), densitometric quantification of the immunoblot was performed using GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0192] Results: Figure 1The results of the experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The IDE protein levels in the brain lysates of the TG2576 (G2), CT10 (G3), and CT10m (G5) groups were significantly increased compared to the control group (G1), and the average level of IDE in CT10m (G5) was higher than that in TG2576 (G2). *p < 0.05, **p < 0.01. Data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one-way ANOVA followed by Dunnett's test. The results indicate that CT10 and CT10m treatments increase IDE levels and their clearance of Aβ peptides, thereby showing a neuroprotective effect.

[0193] Example 2 : To evaluate the efficacy of the microbiome composition in affecting insulin signaling and its upstream molecule NSE in an AD mouse model

[0194] This example provides an evaluation of the efficacy of the microbiome composition (specifically, the CT10, CT10x, and CT10m compositions) in affecting insulin signaling and its upstream molecule NSE related to Akt activation and neuroprotection in an in vivo mouse model of AD.

[0195] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeleton assembly, synapse formation, neurotransmitter and memory functions, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). Neuronal-specific enolase (NSE) is a glycolytic isoenzyme and a highly specific marker of central and peripheral neurons and neuroendocrine cells. NSE is also expressed in microglia (Hafner et al., 2013; Pislar et al., 2017) and astrocytes, especially in reactive astrocytes (Vinores et al., 1985). NSE levels in serum and CSF have been used as biomarkers for injury, cancer, and neurodegenerative diseases (Schmidt et al., 2014; Isgro et al., 2015). The elevation of NSE promotes glycolysis, cell proliferation, activation, and migration through the PI3K / AKT and MAPK / ERK pathways. NSE-mediated PI3K activation also regulates RhoA kinase, which affects the induction of actin cytoskeleton reorganization and neurite growth (Haque et al., 2018).

[0196] Mouse model: The mouse model described in Example 1 was used in this study.

[0197] Study: The Tg2576 mice were divided into five groups of 15 animals each and were provided with either mock (DPBS) or the microbiome compositions (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by oral gavage daily for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. The animals were sacrificed at the end of the study and subsequently subjected to histological analysis.

[0198] Methods: To determine whether CT10, CT10x, or CT10m treatment affects insulin signaling and its upstream molecule NSE, which is related to Akt activation and neuroprotection, the NSE protein levels in brain samples of TG2576 (AD mouse model) were examined by Western blotting. The major parts of the cortex, hippocampus, and thalamus of the brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom SDS-polyacrylamide Bis-tris gel (4% - 12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) protein-free blocking buffer (catalog number 927-80001, LI-COR) or a 5% non-fat dry milk solution and incubated with The NSE-specific antibody (Catalog No. sc-21738, Santa Cruz Biotechnology) diluted 1:1000 in T20 (TBS) Protein-Free Antibody Diluent (Catalog No. 927-85001, LI-COR) was incubated overnight at 4 °C under gentle shaking conditions. The next day, the membranes were washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 h at room temperature in a 1:2000 dilution of HRP-conjugated secondary anti-rabbit antibody (Catalog No. 7074; Cell signaling). After visualization with a G:Box Mini (Syngene), densitometric quantification of the immunoblots was performed by GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0199] Results: Figure 2 The results of this experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The NSE protein levels were significantly increased in the brain lysates of the CT10 (G3) and CT10m (G5) groups compared to TG2576 (G2). **p < 0.01, ***p < 0.001, data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data sets were analyzed by one-way ANOVA followed by Dunnett's test. The results indicate that CT10 and CT10m treatments increase NSE levels and may trigger insulin-Akt signaling activation and neuroprotection.

[0200] Example 3 : To evaluate the efficacy of the microbiome compositions in affecting Akt signaling in an AD mouse model

[0201] This example provides an evaluation of the efficacy of the microbiome compositions (specifically the CT10, CT10m, and CT10x compositions) in affecting Akt signaling in an in vivo mouse model of AD.

[0202] Background:The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synapse formation, neurotransmitter function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). AD is associated with the relative state of insulin resistance in the brain and impaired insulin / IGF-1 expression and Akt signaling. Insulin receptors degenerate and tyrosine kinase activity is reduced (IR desensitization) (Steen E and de la Monte 2005; Frohlich L and Hoyer S. Ann. 1999). AD is associated with reduced insulin-mediated glucose uptake. Insulin levels are decreased in the AD brain and CSF (Frohlich 1998; Craft 1998).

[0203] Mouse model: The mouse model described in Example 1 was used in this study.

[0204] Study: Tg2576 mice were divided into five groups of 15 animals each and were provided with either mock (DPBS) or the microbiome composition (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, followed by histological analysis.

[0205] Methods:To determine whether CT10, CT10x, or CT10m treatment affects Akt signaling, the levels of the active form of Akt, p-Akt(Ser473), and total Akt1 enzyme protein were measured by Western blotting in brain samples from TG2576 mice. The major parts of the cortex, hippocampus, and thalamus of the brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom-made SDS-polyacrylamide Bis-tris gel (4%-12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) protein-free blocking buffer (catalog number 927-80001, LI-COR) or 5% non-fat milk powder solution and incubated overnight at 4 °C with p-Akt(Ser473), Akt1-specific antibody (catalog number 4060, #4691 Cell Signaling Tech.) diluted 1:1000 in T20(TBS) protein-free antibody dilution buffer (catalog number 927-85001, LI-COR) under gentle shaking conditions. The next day, the membrane was washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in a 1:2000 dilution of HRP-conjugated secondary anti-rabbit antibody (catalog number 7074; Cell signaling). After visualization with a G:Box Mini (Syngene), densitometric quantification of the immunoblot was performed using GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0206] Results: Figure 3The results of the experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The protein level of p-Akt(Ser473) in the brain lysates of TG2576 mice was significantly decreased compared to non-Tg mice. The protein level of p-Akt(Ser473) was significantly restored in mice treated with CT10 (G3) or CT10x (G4), CT10m (G5) compared to vehicle-treated TG2576 mice (G2). The total Akt1 protein kinase level in the brain lysates of TG2576 mice treated with CT10 (G3) or CT10x (G4) or CT10m (G5) was increased to approximately 120% of the level in the control group (non-Tg, G1). *p < 0.05 and **p < 0.01 and ***p < 0.001, and ****p < 0.0001. Data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one-way ANOVA, followed by Dunnett's test. The results suggest that CT10, CT10x, and CT10m treatments increase and restore the insulin / Akt signaling pathway, contributing to neuronal survival and memory and other brain functions.

[0207] Example 4 : To evaluate the efficacy of the microbiome composition in affecting insulin receptor β levels in an AD mouse model

[0208] This example provides an evaluation of the efficacy of the microbiome composition, particularly the CT10, CT10m, and CT10x compositions, in affecting insulin receptor β levels in an in vivo mouse model of AD.

[0209] Mouse model: The mouse model described in Example 1 was used in this study.

[0210] Study: Tg2576 mice were divided into five groups of 15 animals each and were provided with mock (DPBS) or the microbiome composition (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with CT10 composition; (iv) G4: Tg2576 transgenic mice treated with CT10x composition; and (v) G5: Tg2576 transgenic mice treated with CT10m composition. Animals were sacrificed at the end of the study, followed by histological analysis.

[0211] Methods:To confirm whether CT10, CT10x or CT10m treatment affects insulin signaling, the insulin receptor β protein levels in brain samples of TG2576 mice were measured by Western blotting. The major parts of the cortex, hippocampus and thalamus of the brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom-made SDS-polyacrylamide Bis-tris gel (4%-12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) protein-free blocking buffer (catalog number 927-80001, LI-COR) or 5% non-fat milk powder solution and incubated overnight at 4 °C with a 1:1000 dilution of insulin receptor β-specific antibody (catalog number 3025, Cell Signaling Tech.) under shaking conditions. The next day, the membrane was washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in a 1:2000 dilution of HRP-conjugated secondary anti-rabbit antibody (catalog number 7074; Cell signaling). After visualization using a G:Box Mini (Syngene), densitometric quantification of the immunoblot was performed by GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0212] Results: Figure 4 The results of this experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The average level of insulin receptor β in TG2576 was decreased compared to the control group (non-Tg), but not significantly. The insulin receptor β protein level in the brain lysates of the CT10m (G5) group was significantly increased compared to TG2576 (G2). **p < 0.01. Data are presented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one-way ANOVA followed by Dunnett's test. The results indicate that CT10m treatment increases and restores insulin / Akt signaling pathway by increasing the insulin receptor β level.

[0213] Example 5 : To evaluate the efficacy of the microbiome composition in affecting the level of glucose transporter 3 in an AD mouse model

[0214] This example provides an evaluation of the efficacy of the microbiome composition (specifically, CT10, CT10m, and CT10x compositions) in affecting the level of glucose transporter 3 in an in vivo mouse model of AD.

[0215] Mouse model: The mouse model described in Example 1 was used in this study.

[0216] Study: Tg2576 mice were divided into five groups of 15 animals each and were provided with either a mock (DPBS) or the microbiome composition (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, followed by histological analysis.

[0217] Methods: To confirm whether CT10, CT10x, or CT10m treatment affects insulin signaling and its downstream targets, the protein level of glucose transporter 3 (Glut3) in brain samples of TG2576 mice was measured by Western blotting. The major parts of the cortex, hippocampus, and thalamus of the brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom SDS-polyacrylamide Bis-tris gel (4% - 12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. Using (TBS) Block the membrane with Protein-Free Blocking Buffer (Catalog No. 927 - 80001, LI - COR) or 5% non - fat milk powder solution for 1 hour, and incubate it overnight at 4 °C under shaking conditions with a 1:1000 - diluted Glucose Transporter 3 (Glut3) - specific antibody (Catalog No. ab191071, abcam.). The next day, wash the membrane thoroughly in TBST (0.1% Tween 20) and incubate it for 1 hour at room temperature in a 1:2000 - diluted HRP - conjugated secondary anti - rabbit antibody (Catalog No. 7074; Cell signaling). After visualization with G:Box Mini (Syngene), perform densitometric quantification of the immunoblot by GeneTools (Syngene). Normalize the target bands using their respective β - actin loading controls.

[0218] Results: Figure 5 The results of this experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The average level of Glut3 in TG2576 (G2) is decreased compared to the control group (non - Tg), but not significantly. The Glut3 protein level in the brain lysates of the CT10m (G5) group is significantly increased compared to TG2576 (G2). *p < 0.05. Data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one - way ANOVA followed by Dunnett's test. The results indicate that CT10m treatment increases and restores insulin / Akt signaling pathway and energy metabolism by increasing the Glucose Transporter 3 (Glut3) protein level.

[0219] Example 6 : Evaluate the efficacy of the microbiome composition in affecting the level of the protein RBAP48 in an AD mouse model

[0220] This example provides an evaluation of the efficacy of the microbiome composition (specifically the CT10, CT10m, and CT10x compositions) in affecting the level of the protein RBAP48 related to memory function in an in - vivo mouse model of AD.

[0221] Background:The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeleton assembly, synapse formation, neurotransmitter and memory functions, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). RBAP48 interacts with histones and modifies histone acetylation, which is crucial for memory consolidation. Loss of RBAP48 is a key to age-related memory decline (Pavlopoulos et al. 2013). RBAP48 interacts with the complex of CREB-binding protein and phosphorylated CREB and is involved in learning and memory (Zhang et al., 2000).

[0222] Mouse model: The mouse model described in Example 1 was used in this study.

[0223] Study: Tg2576 mice were divided into five groups of 15 animals each and were provided with mock (DPBS) or the microbiome composition (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, followed by histological analysis.

[0224] Methods:To determine whether CT10, CT10x, or CT10m treatment affects insulin signaling and its downstream target molecules related to memory function, the protein level of RBAP48 in brain samples of TG2576 (AD mouse model) was measured by Western blotting. The major parts of the cortex, hippocampus, and thalamus of brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom SDS-polyacrylamide Bis-tris gel (4%-12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) protein-free blocking buffer (catalog number 927-80001, LI-COR) or 5% non-fat milk powder solution and incubated overnight at 4 °C under shaking conditions with a 1:1000 dilution of an RBAP48-specific antibody (catalog number GTX70232, GeneTex). The next day, the membrane was washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in a 1:2000 dilution of an HRP-conjugated secondary anti-mouse antibody (catalog number 7076; Cell signaling). After visualization with a G:Box Mini (Syngene), densitometric quantification of the immunoblot was performed using GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0225] Results: Figure 6Show the results of this experiment. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The RBAP48 protein levels were significantly increased in the brain lysates of the CT10 (G3), CT10x (G4), and CT10m (G5) groups compared to TG2576 (G2). *p < 0.05, ****p < 0.0001. Data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one-way ANOVA followed by Dunnett's test. The results indicate that CT10, CT10x, and CT10m treatments increase and restore insulin / Akt signaling pathway and its downstream memory function targets by increasing the RBAP48 protein level.

[0226] Example 7 : Evaluate the efficacy of the microbiome composition in affecting the levels of p-4EBP1, Akt, and mTOR in an AD mouse model

[0227] This example provides an evaluation of the efficacy of the microbiome composition, particularly the CT10, CT10m, and CT10x compositions, in affecting the levels of p-4EBP1, Akt, and mTOR in an in vivo mouse model of AD.

[0228] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeleton assembly, synapse formation, neurotransmission, and memory function, as well as plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). Hyperphosphorylation of 4EBP1 leads to the activation of cap-dependent translation and protein synthesis (Pause et al., 1994). Both the PI3 kinase / Akt pathway and the mTOR kinase regulate 4EBP1 activity (Brunn et al., 1997. Gingras et al., 1998). The level of phosphorylated phospho-4EBP1 (Thr37 / 46) is decreased in the hippocampus of the Tg2576 mouse brain. Inhibition of mTOR signaling in an AD mouse model is associated with impairment of synaptic plasticity (Ma et al., 2010).

[0229] Mouse model: The mouse model described in Example 1 was used in this study.

[0230] Study:Tg2576 mice were divided into five groups of 15 animals each and were provided with either mock (DPBS) or the microbiome compositions (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) via daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study and subsequently subjected to histological analysis.

[0231] Methods: To determine whether CT10, CT10x, or CT10m treatment affects insulin signaling and its downstream target molecules related to protein synthesis and translation, the protein level of phospho-4EBP1 (Thr37 / 46) in brain samples of TG2576 (AD mouse model) was measured by Western blotting. The major parts of the cortex, hippocampus, and thalamus of brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (catalog number 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom SDS-polyacrylamide Bis-tris gel (4% - 12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (catalog number IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) protein-free blocking buffer (catalog number 927-80001, LI-COR) or a 5% non-fat dry milk solution and incubated with The phospho-4EBP1 (Thr37 / 46) specific antibody (Catalog No. 2855, Cell signaling) diluted 1:1000 in T20 (TBS) Protein-Free Antibody Diluent (Catalog No. 927-85001, LI-COR) was incubated overnight at 4 °C under gentle shaking conditions. The next day, the membranes were washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in a 1:2000 dilution of HRP-conjugated secondary anti-rabbit antibody (Catalog No. 7074; Cell signaling). After visualization with a G:Box Mini (Syngene), densitometric quantification of the immunoblots was performed by GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0232] Results: Figure 7 The results of this experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The phospho-4EBP1 (Thr37 / 46) protein levels were significantly increased in the brain lysates of the CT10x (G4) and CT10m (G5) groups compared to TG2576 (G2). **p < 0.01, ****p < 0.0001. Data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one-way ANOVA, followed by Dunnett's test. The results indicate that CT10x and CT10m treatments increase insulin / Akt signaling pathway and its downstream protein synthesis targets by increasing phospho-4EBP1 (Thr37 / 46) protein levels.

[0233] Example 8 : To evaluate the efficacy of the microbiome composition in affecting the level of the antioxidant response regulator NRF2 in an AD mouse model

[0234] This example provides an evaluation of the efficacy of the microbiome composition, particularly the CT10, CT10m, and CT10x compositions, in affecting the level of the antioxidant response regulator NRF2 in an in vivo mouse model of AD.

[0235] Background:The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeleton assembly, synapse formation, neurotransmitter and memory functions, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). NRF2 (nuclear factor erythroid 2-related factor 2) is the main regulator of the cellular antioxidant response, regulating the expression of more than 200 genes containing antioxidant response elements (AREs). The dietary herbal medicine (Bungeanum) improves cognitive dysfunction and neurological deficits in an aging mouse model by activating the PI3K / Akt / NRF2 signaling pathway (Zhao et al., 2020). NRF2 protects neuronal cells from Aβ-mediated oxidative and metabolic damage through activation of the PI3K / GSK-3 axis (Sotolongo et al., 2020). Activation of the NRF2 / ARE pathway alleviates cognitive dysfunction in the Tg mouse AD model by regulating oxidative stress (Tian et al., 2018).

[0236] Mouse model: The mouse model described in Example 1 was used in this study.

[0237] Study: Tg2576 mice were divided into five groups of 15 animals each and were provided with either mock (DPBS) or the microbiome composition (CT10, CT10m, or CT10x; composition details are listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS; (ii) G2: Tg2576 transgenic mice treated with DPBS; (iii) G3: Tg2576 transgenic mice treated with the CT10 composition; (iv) G4: Tg2576 transgenic mice treated with the CT10x composition; and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, followed by histological analysis.

[0238] Methods:To determine whether CT10, CT10x, or CT10m treatment affects insulin signaling and its downstream target molecules related to antioxidant response and neuroprotection, the NRF2 protein level in brain samples of TG2576 (AD mouse model) was measured by Western blotting. The major parts of the cortex, hippocampus, and thalamus of brain lysates were obtained by immersing the tissues in PhosphoSafe buffer (EMD Millipore) and lysing them in tubes containing lysis matrix D (MP Biomedicals). After centrifugation of the samples, the concentration of the protein supernatant was determined by the Bradford method using BioRad protein assay reagent (Catalog No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer and boiled at 70 °C for 10 minutes. The samples were run on a custom-made SDS-polyacrylamide Bis-tris gel (4%-12%, Invitrogen) using MES running buffer and then transferred to a PVDF membrane (Catalog No. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using (TBS) Protein-free blocking buffer (Catalog No. 927-80001, LI-COR) or 5% non-fat dry milk solution and incubated overnight at 4 °C with gentle shaking with an NRF2-specific antibody (Catalog No. 12721, Cell signaling) diluted 1:1000 in T20 (TBS) Protein-free antibody dilution buffer (Catalog No. 927-85001, LI-COR). The next day, the membrane was washed thoroughly in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in a 1:2000 dilution of HRP-conjugated secondary anti-rabbit antibody (Catalog No. 7074; Cell signaling). After visualization with a G:Box Mini (Syngene), densitometric quantification of the immunoblot was performed by GeneTools (Syngene). The target bands were normalized using their respective β-actin loading controls.

[0239] Results: Figure 8The results of the experiment are shown. Each point in groups 1, 4, and 5 represents pooled brain lysates from two animals. The NRF2 protein levels were significantly increased in the brain lysates of the CT10 (G3), CT10x (G4), and CT10m (G5) groups compared to TG2576 (G2). *p < 0.05, **p < 0.01. Data are represented as mean ± SEM. Statistical analysis was performed using GraphPad Prism. The data set was analyzed by one-way ANOVA followed by Dunnett's test. The results indicate that CT10, CT10x, and CT10m treatments increase insulin / Akt signaling pathway and its downstream antioxidant response and neuroprotection targets by increasing NRF2 protein levels.

[0240] Other embodiments

[0241] Those skilled in the art will understand that various changes, modifications, and improvements to the present disclosure will readily occur to them. Such changes, modifications, and improvements are intended to be part of the present disclosure and are intended to be within the spirit and scope of the invention. Accordingly, the foregoing description and drawings are by way of example only, and any invention described in the present disclosure is further described in detail by the appended claims.

[0242] Those skilled in the art will understand the typical deviations or error criteria attributable to values obtained in assays or other procedures as described herein. Publications, websites, and other reference materials cited herein to describe the background of the invention and to provide additional details regarding its practice are hereby incorporated by reference in their entirety.

[0243] It should be understood that while the embodiments of the present invention have been described in connection with their specific embodiments, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0244] Equivalent cases

[0245] Those skilled in the art will recognize or be able to ascertain many equivalent embodiments of the specific embodiments of the invention described herein using only routine experimentation. The scope of the present invention is not intended to be limited to the above specification, but rather as set forth in the following claims:

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280] Appendix 1-3 Target Metabolites

[0281] C10H10O3S

[0282] C32H30O12

[0283] C6H10O8

[0284] 1-Methyl-4-imidazoleacetic acid

[0285] 1-Methyladenosine

[0286] 1-Methylhistamine

[0287] 1-Methylhistidine

[0288] 3-Methylhistidine

[0289] 1-Methylhistidine; 3-Methylhistidine

[0290] 1-Methylnicotinamide

[0291] 1-Palmitoyl-glycero-3-phosphocholine-1

[0292] 1,2-Dipalmitoyl-glycero-3-phosphoethanolamine-1

[0293] 1,2-Dipalmitoyl-glycero-3-phosphoethanolamine-2

[0294] 11-Aminoundecanoic acid

[0295] 15(S)-HETE

[0296] 17α-Hydroxyprogesterone

[0297] 17α-Hydroxyprogesterone-2

[0298] Deoxycorticosterone-2

[0299] 1H-Imidazole-4-propionic acid

[0300] 1H-Imidazole-4-propionic acid; 1-Methyl-4-imidazoleacetic acid

[0301] 2-(β-D-Glucosyl)-sn-glycerol

[0302] 2-Aminoadipic acid

[0303] 2-Aminoisobutyric acid 2-Aminobutyric acid

[0304] 2-Arachidonoylglycerol

[0305] 2-Diethylaminoethanol

[0306] 2-Hydroxy-4-methylpentanoic acid

[0307] 2-Hydroxybutyric acid

[0308] 2-Hydroxyglutaric acid

[0309] 2-Hydroxyisobutyric acid

[0310] 2-Hydroxypentanoic acid

[0311] 2-Keto-valeramide acid

[0312] 2-Methylserine

[0313] 2-Oxoarginine

[0314] 2-Oxoglutaric acid

[0315] 2-Oxo-isovaleric acid

[0316] 2-Oxo-isovaleric acid; 2-Oxopentanoic acid

[0317] 2-Phosphoglyceric acid

[0318] 2-Phosphoglyceric acid; 3-Phosphoglyceric acid

[0319] 2-Thiopheneacetic acid

[0320] 2,3-Diphosphoglyceric acid

[0321] 2,6-Diaminopimelic acid

[0322] 2'-Deoxycytidine

[0323] 2’-Deoxyuridine

[0324] 20α-Hydroxyprogesterone

[0325] 21-Deoxycortisol-2

[0326] 21-Hydroxypregnenolone

[0327] 3-(4-Hydroxyphenyl)propanoic acid

[0328] 2-(4-Hydroxyphenyl)propanoic acid

[0329] 3-(3-Hydroxyphenyl)propanoic acid

[0330] 3-(4-Hydroxyphenyl)propionic acid; 2-(4-hydroxyphenyl)propionic acid; 3-(3-hydroxyphenyl)propionic acid; tropic acid; 3-(2-hydroxyphenyl)propionic acid; m-ethoxybenzoic acid; 3-phenyllactic acid; p-methoxyphenylacetic acid

[0331] 3-cis-Hydroxy-β,ε-carotene-3'-one

[0332] 3-Guanidinopropionate

[0333] 3-Hydroxy-2-methyl-4-pyrone

[0334] 3-Hydroxy-3-methylglutaric acid

[0335] 3-Hydroxy-3',4'-dehydro-β,γ-carotene

[0336] 3-Hydroxybutyric acid

[0337] 3-Hydroxybutyric acid; 2-hydroxybutyric acid; 2-hydroxyisobutyric acid

[0338] 3-Hydroxybutyric acid; 2-hydroxyisobutyric acid

[0339] 3-Hydroxyglutaric acid

[0340] 3-Hydroxytetradecanoic acid-1

[0341] 3-Indoxyl sulfate

[0342] 3-Mercaptoacetic acid

[0343] 3-Methylhistamine; 1-methylhistamine

[0344] 3-Methylhistidine; 1-methylhistidine

[0345] 3-Oxocholic acid

[0346] 3-Phosphoglyceric acid

[0347] 3-Ureidopropionic acid

[0348] 3,4-Dihydroxyhydrocinnamic acid; homovanillic acid; hydroxyphenyllactic acid

[0349] 4-Acetamidobutyric acid

[0350] 4-Guanidinobutyric acid

[0351] 4-Hydroxyquinoline

[0352] 4-Methyl-2-oxopentanoic acid

[0353] 3-Methyl-2-oxopentanoic acid

[0354] 2-Oxohexanoic acid

[0355] 4-Methyl-2-oxopentanoic acid; 3-Methyl-2-oxopentanoic acid; 2-Oxohexanoic acid

[0356] 4-Oxopyrrolidine-2-carboxylic acid

[0357] 5-Amino-4-hydroxynaphthalene-1,3-disulfonic acid

[0358] 5-Amino-4-oxopentanoic acid

[0359] 5-Hydroxyindoleacetic acid

[0360] 5-Hydroxylysine

[0361] 5-Hydroxypentanoic acid; 2-Hydroxypentanoic acid

[0362] 5-Hydroxytryptophan

[0363] 5-Isopropyl-2'-deoxyuridine triphosphate

[0364] 5-Methoxyindoleacetic acid; Indole-3-lactic acid

[0365] 5-Methyl-2'-deoxycytidine

[0366] 5-Oxoproline

[0367] 5α-Cholestan-3-one-1

[0368] 5α-Cholestan-3-one-2

[0369] 5α-Pregnane-3,20-dione

[0370] 6-Oxohexadecanoic acid

[0371] 6-Hydroxyhexanoic acid; 2-Hydroxy-4-methylpentanoic acid

[0372] 6-Hydroxynicotinic acid

[0373] 7-Dehydrocholesterol

[0374] 7-Dehydrocholesterol

[0375] 7-Dehydrocholesterol-2

[0376] Lathosterol-2

[0377] 7-Dehydrocholesterol-3

[0378] Lathosterol-3

[0379] 7-Methoxy-2-methylisoflavone

[0380] 7-Methylguanine

[0381] 7-Methylguanine; 3-Methylguanine

[0382] 7,8-dihydrobiopterin

[0383] 7,8-dihydrobiopterin

[0384] abietic acid

[0385] abietic acid-1

[0386] abietic acid-3

[0387] AC(10:0)

[0388] AC(12:0)-1

[0389] AC(12:0)-2

[0390] AC(12:1)

[0391] AC(12:1)-1

[0392] AC(12:1)-3

[0393] AC(13:1)

[0394] AC(13:1)-1

[0395] AC(14:0)-1

[0396] AC(14:0)-2

[0397] AC(14:1)-1

[0398] AC(14:1)-2

[0399] AC(14:1)-3

[0400] AC(14:1)-4

[0401] AC(14:2)-1

[0402] AC(14:2)-2

[0403] AC(14:2)-3

[0404] AC(14:3)-1

[0405] AC(14:3)-2

[0406] AC(14:3)-3

[0407] AC(14:3)-4

[0408] AC(15:0)-1

[0409] AC(15:0)-2

[0410] AC(16:1)

[0411] AC(16:2)-1

[0412] AC(16:2)-2

[0413] AC(17:0)-1

[0414] AC(17:0)-2

[0415] AC(17:1)

[0416] AC(18:0)

[0417] AC(18:1)

[0418] AC(18:2)-1

[0419] AC(18:2)-2

[0420] AC(20:0)

[0421] AC(20:1)

[0422] AC(22:0)

[0423] Hydroxamic acid acetate; Gly

[0424] ADMA; SDMA

[0425] ADP

[0426] ADP-ribose

[0427] AEA(22:6)

[0428] Ala

[0429] ala ser / gly thr

[0430] Aminoacetone

[0431] AMP

[0432] AMP; dGMP

[0433] Anandamide

[0434] ANDS(C-SCOPE IS)

[0435] Anserine

[0436] Arachidic acid

[0437] Arachidonic acid

[0438] Arg

[0439] Argininosuccinic acid

[0440] Ascorbic acid

[0441] Madecassic acid

[0442] Madecassic acid-1

[0443] Asn

[0444] Asp

[0445] Asp Asp Pro Ser

[0446] Asp Gly His Asp

[0447] Asp Leu Asn Arg

[0448] Asp-Pro

[0449] ATP

[0450] Baicalein

[0451] Behenic acid

[0452] Betaine

[0453] Betaine aldehyde_+H2O

[0454] Betulinic acid

[0455] Betulinic acid-2

[0456] Biopterin

[0457] Biotin

[0458] Butyrobetaine

[0459] C; C

[0460] C3H8N4O

[0461] C4H7NO4

[0462] C5H12N2O2

[0463] C5H6O7

[0464] C6H10O8

[0465] C6H11NO2

[0466] C6H12N2O3

[0467] C7H9N3O2

[0468] C8H17NO

[0469] C8H18N2O3

[0470] C9H18N2O

[0471] cAMP

[0472] Campesterol

[0473] Carbachol

[0474] Carboxymethyllysine

[0475] Carnitine

[0476] Carnosine

[0477] Carnosine; His-Ala; Ala-His

[0478] Chenodeoxycholic acid

[0479] Cholesterol

[0480] Cholesterol sulfate

[0481] Choline cholate

[0482] Cis-11-eicosenoic acid

[0483] Cis-11,14-eicosadienoic acid-1

[0484] Cis-11,14-eicosadienoic acid-2

[0485] Cis-4,7,10,13,16,19-docosahexaenoic acid

[0486] Cis-5,8,11,14,17-eicosapentaenoic acid

[0487] Cis-8,11,14-eicosatrienoic acid

[0488] Cis-aconitic acid

[0489] Citric acid

[0490] Citrulline

[0491] Corosolic acid

[0492] Corticosterone

[0493] Cortisone

[0494] 21-Deoxycortisol-1

[0495] Cortisol

[0496] 18-Hydroxycorticosterone

[0497] Humulone

[0498] Creatine

[0499] Creatinine

[0500] Tiglic acid

[0501] CSA; CSA

[0502] Cyclodopaglucoside

[0503] Cyclohexylamine

[0504] Cys Cys Csy Asn Asn

[0505] Cystathionine

[0506] Cysteine glutathione disulfide

[0507] Cystine

[0508] Cytidine

[0509] Cytosine

[0510] Daidzein

[0511] Dansulfonic acid (C - SCOPE IS)

[0512] Deoxycholic acid

[0513] Desthiobiotin

[0514] dGDP; ADP

[0515] dGTP; ATP

[0516] Diethanolamine

[0517] Dimethylaminoethanol

[0518] DOPA

[0519] DPA; DPA

[0520] Diprophylline

[0521] Ectoine

[0522] Erucic acid

[0523] Ethanolamine

[0524] Phosphoethanolamine

[0525] Ethyl arachidonate

[0526] Ethyl glucuronide

[0527] FA16:1

[0528] FAC18:1

[0529] FA(12:0)

[0530] FA(13:0)

[0531] FA(14:1)

[0532] FA(14:l)-2

[0533] FA(14:2)-1

[0534] FA(14:2)-2

[0535] FA(14:3)

[0536] FA(15:0)

[0537] FA(15:0)-1

[0538] FA(15:1)

[0539] FA(15:1)-2

[0540] FA(16:2)-1

[0541] FA(16:2)-2

[0542] FA(16:3

[0543] FA(16:3)-2

[0544] FA(17:0)

[0545] FA(17:1)

[0546] FA(17:2)

[0547] FA(17:3)

[0548] FA(19:0)

[0549] FA(19:0)-1

[0550] FA(19:1)

[0551] FA(19:2)

[0552] FA(20:3)

[0553] FA(22:2)

[0554] FA(22:3)-1

[0555] FA(22:3)-2

[0556] FA(22:4)-1

[0557] FA(22:4)-2

[0558] FA(22:5)-1

[0559] FA(22:5)-2

[0560] FA(24:0)

[0561] FA(24:2)

[0562] FA(24:4)

[0563] FA(24:5)-1

[0564] FA(24:5)-2

[0565] Flavanone

[0566] Formiminoglutamic acid

[0567] Formononetin

[0568] Fucosyltryptophan

[0569] Fucosyl-lysine

[0570] Fumaric acid

[0571]

[0572] GABA

[0573] GABA; 3-Aminoisobutyric acid

[0574] Galactosylhydroxylysine

[0575] Galacturonic acid-1

[0576] Glucuronic acid-1

[0577] Galacturonic acid; Glucuronic acid

[0578] γ-Glu-Gln

[0579] GDP

[0580] Genistein

[0581] Gln

[0582] Glu

[0583] Glu; Isoglutamic acid; N-Methylaspartic acid; N-Acetylserine

[0584] Glucaric acid

[0585] Gluconic acid

[0586] Glucono-δ-lactone

[0587] Glucosamine

[0588] Glucosamine 6-sulfate

[0589] Glucose 6-phosphate

[0590] Glucosyl-glycerol

[0591] Glutaric acid

[0592] Methylsuccinic acid

[0593] Glutaric acid; Methylsuccinic acid

[0594] Glutathione (GSSG)_divalent

[0595] Gly

[0596] Gly Lys

[0597] Gly-Ala

[0598] Gly-Asp

[0599] Gly-Asp; Asp-Gly

[0600] Gly-Leu; N-acetyllysine; Val-Ala; Ala-Val; Leu-Gly

[0601] Glyceric acid

[0602] Glycerol

[0603] Glycerol 3-phosphate

[0604] Glycerophosphocholine

[0605] Daidzein

[0606] Glycochenodeoxycholic acid

[0607] Glycocholic acid

[0608] Glycodeoxycholic acid

[0609] Glyoxylic acid

[0610] GMP

[0611] Guanidinosuccinic acid

[0612] Guanidinoacetic acid

[0613] L-Gulono-1,4-lactone; Glucono-1,5-lactone

[0614] Agavogenin

[0615] Heneicosanoic acid

[0616] 19-Methylarachidic acid

[0617] Heptadecanoic acid-1

[0618] FA(17:0)-1

[0619] Heptadecanoic acid-2

[0620] FA(17:0)-2

[0621] Enanthic acid

[0622] Hexanoic acid

[0623] Hippuric acid

[0624] Hippuric acid (benzylglycine)

[0625] His

[0626] His Pro Ser Val Arg Tyr Thr

[0627] His-Asp

[0628] Histamine

[0629] Homoarginine

[0630] Glycine betaine

[0631] Carnosine

[0632] Homocitrulline

[0633] Cysteine sulfinic acid

[0634] Homoserine

[0635] Homovanillic acid

[0636] Hydroxyphenyl lactic acid

[0637] Hydroxyindole

[0638] Hydroxyoctanoic acid

[0639] Hydroxyprogesterone caproate

[0640] Hydroxyproline

[0641] Hydroxytetradecanoic acid

[0642] Hyodeoxycholic acid

[0643] Taurine

[0644] IDP

[0645] Ile

[0646] Ile; Leu; Alloisoleucine

[0647] Ile; Leu; Alloisoleucine; 6-Aminohexanoic acid

[0648] Imidazole-4-acetic acid

[0649] Imidazole lactic acid

[0650] IMP

[0651] Indole-3-acetic acid

[0652] Indole-3-carbaldehyde

[0653] Indole-3-lactic acid-1

[0654] 5-Methoxyindoleacetic acid-1

[0655] Indole-3-lactic acid; 5-Methoxyindoleacetic acid

[0656] Indole-3-propionic acid

[0657] Indole-3-propionic acid (IPA)

[0658] Inosine 2',3'-cyclophosphate clMP

[0659] 2-Hydroxyethanesulfonic acid

[0660] Isobutyryl carnitine; Butyryl carnitine

[0661] Isocitric acid

[0662] Isoglutamic acid

[0663] Isoliquiritigenin-1

[0664] Isoliquiritigenin-2

[0665] Isoliquiritigenin-3

[0666] Isonicotinamide; Nicotinamide

[0667] Isovalerylalanine-2

[0668] N-Acetylleucine-2

[0669] Isovalerylalanine; N-Acetylleucine

[0670] Isovaleryl carnitine

[0671] Kynurenic acid

[0672] Kynurenine

[0673] Lactamide

[0674] Lactic acid

[0675] Lanosterol

[0676] Lauric acid

[0677] Leu

[0678] Leukotriene B4

[0679] Linoleic acid

[0680] Linolenic acid

[0681] Linoleoyl ethanolamide

[0682] Liquiritigenin

[0683] Lithocholic acid

[0684] Luteolin

[0685] Lys

[0686] Lys - Asp

[0687] Lys - Val

[0688] Malic acid

[0689] Mannosamine

[0690] MCA

[0691] Met

[0692] Methionine sulfone

[0693] Methionine sulfoxide

[0694] Methylmalonic acid; Succinic acid

[0695] Morpholine

[0696] Mucic acid

[0697] Myristic acid

[0698] Myristic acid 14:0

[0699] Myristoleic acid

[0700] N-(1 - deoxy - 1 - fructosyl)valine

[0701] N - Acetyl - β - alanine

[0702] N - Acetyl - β - alanine

[0703] N - Acetylalanine

[0704] N - Acetylalanine;

[0705] N - Acetylasparagine

[0706] N - Acetylaspartic acid

[0707] N - Acetylgalactosamine; N - Acetylglucosamine; N - Acetylmannosamine

[0708] N - Acetylglucosamine

[0709] N - Acetylglutamic acid

[0710] N-acetylglutamine

[0711] N-acetylglycine

[0712] N-acetylhistidine

[0713] N-acetylleucine

[0714] N-acetyllysine

[0715] N-acetylmethionine

[0716] N-acetylneuraminic acid

[0717] N-acetylornithine

[0718] N-acetylphenylalanine

[0719] N-acetyltryptophan

[0720] N-acetyltyrosine

[0721] N-carbamylglutamate

[0722] N-carboxymethylserine

[0723] N-ethylmaleimide + H2O

[0724] N-formylaspartic acid

[0725] N-formylglycine

[0726] N-formylmethionine

[0727] N-glycolylneuraminic acid

[0728] N-hydroxy-L-tryptophan

[0729] N-methylethanolamine phosphate

[0730] N-methylproline

[0731] N,N-dimethylglycine

[0732] N'-formylkynurenine

[0733] N1-acetylspermidine

[0734] N1-acetylspermidine; N8-acetylspermidine

[0735] N1-methyl-4-pyridone-5-carboxamide

[0736] N1-methylguanosine

[0737] N5-ethylglutamine

[0738] N5-Ethylglutamine; N-Acetylornithine

[0739] N6-Acetyllysine

[0740] N6-Methyl-2'-deoxyadenosine

[0741] N6-Methyllysine

[0742] N6,N6,N6-Trimethyllysine

[0743] Naringenin

[0744] Nervonic acid

[0745] Neutral glycosphingolipidyl carnitine propyl betaine (triethylamine)

[0746] Nf-Formylkynurenine

[0747] Nicotinamide

[0748] No match

[0749] Norophthalmic acid

[0750] Norvaline; 2-Amino-2-methylbutyric acid; 5-Aminopentanoic acid; Val

[0751] Nω-Methylarginine

[0752] O-Acetylcarnitine

[0753] O-Acetylhomoserine

[0754] o-Coumaric acid

[0755] p-Coumaric acid

[0756] o-Hydroxybenzoic acid

[0757] Oleanolic acid

[0758] Oleic acid

[0759] Oleoylethanolamide

[0760] AEA(18:1)

[0761] Ophthalmic acid

[0762] Ornithine

[0763] Orotidine; Uridine; Pseudouridine

[0764] p-Hydroxyphenylpyruvic acid

[0765] p-Hydroxyphenylpyruvic acid; Caffeic acid

[0766] Palmitic acid

[0767] Palmitoleic acid

[0768] Palmitoyl carnitine

[0769] Palmitoylethanolamide

[0770] Pantothenic acid

[0771] Penicillamine

[0772] Penicillamine; Met

[0773] Pentadecanoic acid

[0774] Phe

[0775] Phe Met His Glu

[0776] Phe Phe Trp Trp

[0777] Phe-Thr

[0778] Phenylacetylglycine

[0779] Phenol

[0780] Phenyl sulfate

[0781] Phenylpyruvic acid

[0782] Phosphocreatine

[0783] Phosphoenolpyruvate

[0784] Phosphocholine

[0785] Pyridinecarboxylic acid

[0786] Pipecolic acid

[0787] Pipecolic acid; N-methylproline; 1-aminocyclopentanecarboxylic acid

[0788] Piperidine

[0789] Pro

[0790] Pro-Gly; Gly-Pro

[0791] Progesterone

[0792] Proline betaine

[0793] Propionyl carnitine

[0794] XC0061

[0795] Propionyl carnitine; XC0061

[0796] Prostaglandin E1-1

[0797] Prostaglandin D1-1

[0798] Prostaglandin E1-2

[0799] Prostaglandin D1-2

[0800] Putrescine

[0801] Pyridoxal

[0802] Pyrrolidine

[0803] Pyruvic acid

[0804] Retinol Vit A

[0805] Retinol-2

[0806] Riboflavin

[0807] Ribose 5-phosphate

[0808] Ribulose 5-phosphate

[0809] Ribulose 5-phosphate; Ribose 1-phosphate; Xylulose 5-phosphate

[0810] Ricinoelaidic acid

[0811] Ricinoelaidic acid 18:1 hydroxy

[0812] Ricinoelaidic acid-2

[0813] Ricinoelaidic acid-3

[0814] S-acetyl-dihydrolipoamide (XC0086)

[0815] S-adenosylhomocysteine

[0816] S-adenosylmethionine

[0817] S-carboxymethylcysteine

[0818] S-methylcysteine

[0819] S-methylglutathione

[0820] S-methylmethionine

[0821] S-sulfocysteine

[0822] Sarcosine

[0823] SDMA

[0824] Sedoheptulose 7-phosphate Ser

[0825] Ser

[0826] Ser Ala / Thr gly

[0827] Ser Glu Pro Thr Asp Pro

[0828] Serotonin

[0829] Sitosterol

[0830] Spermidine

[0831] Spermine

[0832] Dihydrosphingosine

[0833] Sphingomyelin (d18:1 / 16:0)-1

[0834] Sphingomyelin (d18:1 / 16:0)-2

[0835] Sphingomyelin (d18:1 / 18:0)-1

[0836] Sphingomyelin (d18:1 / 18:0)-2

[0837] Sphingosine

[0838] Stearic acid

[0839] Stearic tetraenoic acid

[0840] Stearoylethanolamide

[0841] Stigmasterol-1

[0842] Stigmasterol-2

[0843] Succinic acid

[0844] Sulfaguanidine (C-SCOPE IS)

[0845] Sulfo-lithocholylglycine

[0846] Taurine

[0847] Taurochenodeoxycholic acid

[0848] Taurocholic acid

[0849] Tauroursodeoxycholic acid

[0850] Tauro-lithocholic acid

[0851] Tauroursodeoxycholic acid

[0852] Terephthalic acid

[0853] Theobromine; Aminophylline; Paraxanthine

[0854] Thiamine

[0855] Thiamine phosphate Thr

[0856] Thr

[0857] Thr Ala Ala

[0858] Thr Asp or Ser Glu

[0859] Threo-3-hydroxyaspartic acid

[0860] Threo-3-hydroxyaspartic acid-1

[0861] Threo-3-hydroxyaspartic acid-2

[0862] Threonic acid

[0863] Thymidine

[0864] Thyroxine trans-pentenedioic acid

[0865] Trans-pentenedioic acid; Itaconic acid

[0866] Tricosanoic acid

[0867] Trigonelline

[0868] Tributyrin

[0869] Trimesic acid; Trimesic acid

[0870] Trimethylamine

[0871] Trimethylamine N-oxide

[0872] Trimethylaminoacetone

[0873] Trp

[0874] Tyr

[0875] UDP-galactose

[0876] UDP-glucose

[0877] Uracil

[0878] Urea

[0879] Uric acid

[0880] Uridine

[0881] Uridine; Pseudouridine

[0882] Urocanic acid

[0883] Ursodeoxycholic acid

[0884] Val

[0885] XA0005

[0886] XA0008

[0887] XA0009

[0888] XA0011

[0889] XA0017

[0890] XA0019

[0891] XA0023

[0892] XA0026

[0893] XA0033

[0894] XA0034

[0895] XA0037

[0896] XA0039

[0897] XA0052

[0898] Xanthosine

[0899] XC0016

[0900] XC0039

[0901] XC0040

[0902] XC0047

[0903] XC0049

[0904] XC0054; XC0055;

[0905] XC0056

[0906] XC0060

[0907] XC0063

[0908] XC0064

[0909] XC0065

[0910] XC0067

[0911] XC0070

[0912] XC0075

[0913] XC0088

[0914] XC0094

[0915] XC0101

[0916] XC0103

[0917] XC0107;

[0918] XC0114;

[0919] XC0117

[0920] XC0118

[0921] XC0119

[0922] XC0120

[0923] XC0126

[0924] XC0133

[0925] XC0135

[0926] XC0138

[0927] XC0139

[0928] XC0140

[0929] Zeaxanthin

[0930] α-Tocopherol

[0931] α-Tocopherol acetate

[0932] β-Ala

[0933] β-Estradiol

[0934] 17α-Estradiol

[0935] β-Hydroxyisovaleric acid

[0936] γ-Butyrobetaine

[0937] γ-Glu-Ala

[0938] γ-Glu-Arg

[0939] γ-Glu-Asn

[0940] γ-Glu-Asp

[0941] γ-Glu-Citrulline

[0942] γ-Glu-Gln

[0943] γ-Glu-Glu

[0944] γ-Glu-Gly

[0945] γ-Glu-His

[0946] γ-Glu-Leu

[0947] γ-Glu-Lys

[0948] γ-Glu-Met

[0949] γ-Glu-Ornithine

[0950] γ-Glu-Phe

[0951] γ-Glu-Ser

[0952] γ-Glu-Taurine

[0953] γ-Glu-Thr

[0954] γ-Glu-Trp

[0955] γ-Glu-Tyr

[0956] γ-Glu-Val

[0957] γ-Glu-Val-Gly

[0958] γ-Tocopherol

[0959] Appendix 2. Known-unknown peaks

[0960] "Known-unknown" peaks not annotated based on chemical standards are shown in the "XA~~~~ / XC~~~~" markers in the result table. Among them, several peaks detected from various biological samples are listed in Appendix 2.

[0961]

[0962]

[0963] Positively and negatively charged molecular ions are measured in positive and negative ion modes, respectively.

[0964] Predicted mass values are calculated for monovalent ions.

[0965] Appendix 2. Known-unknown peaks

[0966]

[0967]

[0968]

[0969] Positively and negatively charged molecular ions are measured in positive and negative ion modes, respectively.

[0970] Predicted mass values are calculated for monovalent ions.

[0971] Appendix 2. Known - Unknown Peaks

[0972]

[0973]

[0974]

[0975] Measure positively and negatively charged molecular ions in cation and anion modes respectively

[0976] The predicted mass values are calculated for singly charged ions

[0977] Appendix 2. Known - Unknown Peaks

[0978]

[0979]

[0980] Measure positively and negatively charged molecular ions in cation and anion modes respectively

[0981] The predicted mass values are calculated for singly charged ions

[0982] Appendix 2. Known - Unknown Peaks

[0983]

[0984]

[0985]

[0986] Measure positively and negatively charged molecular ions in cation and anion modes respectively

[0987] The predicted mass values are calculated for singly charged ions

[0988] Appendix 3. Detected Metabolites

[0989]

[0990]

[0991]

[0992]

[0993]

[0994]

[0995]

[0996]

[0997]

[0998]

[0999]

[1000]

[1001]

[1002]

[1003]

[1004]

[1005]

[1006]

[1007]

[1008]

[1009] Appendix 4. Target Metabolites

[1010]

[1011]

[1012]

[1013]

[1014]

[1015]

Claims

1. A method, the method comprising: administering to a subject a composition comprising one or more microbial strains or microbial components, wherein the subject has been diagnosed with an insulin-related disease, disorder or condition, or is at high risk of developing an insulin-related disease, disorder or condition.

2. A method, the method comprising: administering to a subject a composition comprising one or more microbial metabolites, wherein the subject has been diagnosed with an insulin-related disease, disorder or condition, or is at high risk of developing an insulin-related disease, disorder or condition.

3. A method, the method comprising: administering to a subject a composition comprising: (i) one or more microbial strains or microbial components, or (ii) one or more microbial metabolites, wherein the subject has been diagnosed with an insulin-related disease, disorder or condition, or is at high risk of developing an insulin-related disease, disorder or condition.

4. The method according to any one of the preceding claims, wherein the insulin-related disease, disorder or condition is or includes diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperglycemic hyperosmolar state, gestational diabetes, diabetic dermopathy, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes or polycystic ovary syndrome (PCOS).

5. The method according to any one of the preceding claims, wherein the subject is a mammal.

6. The method according to any one of the preceding claims, wherein the subject is a human.

7. The method according to claim 1 or any one of claims 3 - 6, wherein the one or more microbial strains are from a mammalian microbiome.

8. The method according to claim 1 or any one of claims 3 - 7, wherein the one or more microbial strains are from a human microbiome.

9. The method according to claim 8, wherein the human microbiome is the microbiome of the subject.

10. The method according to claim 9, wherein the human microbiome is administered to maintain or modulate the microbiome of the subject.

11. The method according to any one of the preceding claims, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3 or Appendix 4.

12. The method according to any one of the preceding claims, wherein the one or more microbial metabolites are or comprise bile acids.

13. The method according to any one of the preceding claims, wherein the one or more microbial metabolites are or comprise tauroursodeoxycholic acid.

14. The method according to any one of the preceding claims, wherein the one or more microbial components or microbial metabolites are butyrobetaine, theobromine, 4-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro) or any combination thereof.

15. The method according to any one of the preceding claims, wherein the one or more microbial components or microbial metabolites are 4-hydroxyphenylpyruvic acid, ectoine, gramine, N-acetyl-L-phenylalanine, Nε-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrobetaine, β-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenylacetylglycine, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homoarginine, mucic acid, homocysteine sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indolyl sulfate, nicotinamide, N-formylglycine, ureidoethanolate, N-methylproline, glucaric acid, butyrobetaine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenylacetylglycine, diethanolamine, phosphocholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, β-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiazolidine-4-carboxylic acid, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0XA0027 or any combination thereof.

16. The method according to claim 1 or any one of claims 3-15, wherein the one or more microbial strains are or include Acetobacter hansenii, Terrabacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp. or a combination thereof.

17. The method according to any one of claims 1 or 3 - 16, wherein the one or more microbial strains are or include Acetobacter hansenii, Terrabacter glycolicus, a certain species of Coprococcus, Lactobacillus plantarum, a certain species of Veillonella, a certain species of Bifidobacterium, or a combination thereof.

18. The method according to any one of claims 1 or 3 - 17, wherein the one or more microbial strains are or include Acetobacter hansenii, Terrabacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof.

19. The method according to any one of claims 1 or 3 - 18, wherein the one or more microbial strains are or include Bacillus subtilis.

20. The method according to any one of claims 1 or 3 - 19, wherein the composition comprises two or more microbial strains.

21. The method according to any one of claims 1 or 3 - 20, wherein the composition comprises five or more microbial strains.

22. The method according to any one of claims 1 or 3 - 21, wherein the composition comprises ten or more microbial strains.

23. The method according to any one of the preceding claims, wherein the composition is administered locally, orally, subcutaneously, intravenously, intramuscularly, intracerebrally, intrathecally, rectally, ophthalmically, intravitreally, or suprachoroidally.

24. The method according to claim 23, wherein the composition is administered orally.

25. The method according to claim 23, wherein the composition is administered intravenously.

26. The method according to any one of the preceding claims, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection, or eye drop.

27. The method of any one of claims 1 or 3-26, wherein each of the one or more microbial strains is present in an amount of 10 1 Up to 10 15 The concentration of CFU is present in the composition.

28. The method according to any one of claims 1 or 3-27, wherein each of the one or more microbial strains is present in the composition at a concentration of at least 10 6 CFU.

29. A composition for treating insulin - related diseases, disorders, or afflictions, the composition comprising one or more microbial strains or microbial components.

30. A composition for treating insulin - related diseases, disorders, or afflictions, the composition comprising one or more microbial metabolites.

31. The composition according to claim 29, wherein the one or more microbial strains are from the mammalian microbiome.

32. The composition according to claim 29 or 31, wherein the one or more microbial strains are from the human microbiome.

33. The composition according to claim 32, wherein the human microbiome is the microbiome of the subject.

34. The composition according to claim 33, wherein the human microbiome is administered to maintain or regulate the microbiome of the subject.

35. The composition according to any one of claims 29 - 34, wherein one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3, or Appendix 4.

36. The composition according to any one of claims 29 - 35, wherein the one or more microbial metabolites are or comprise bile acids.

37. The composition according to any one of claims 29 - 36, wherein the one or more microbial metabolites are or comprise tauroursodeoxycholic acid.

38. The composition according to any one of claims 29 - 35, wherein one or more microbial components or microbial metabolites are butyrylcarnitine, theobromine, 4-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro) or any combination thereof.

39. The composition according to any one of claims 29 - 35, wherein one or more microbial components or microbial metabolites are 4-hydroxyphenylpyruvic acid, ectoine, gramine, N-acetyl-L-phenylalanine, Nε-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine, b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenylacetylglycine, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homoarginine, mucic acid, homocysteine sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indolylsulfate, nicotinamide, N-formylglycine, ureidoethanolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenylacetylglycine, diethanolamine, phosphocholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiazolidine-4-carboxylic acid, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0XA0027 or any combination thereof.

40. The composition according to any one of claims 29 or 31 - 39, wherein the one or more microbial strains are or include Acetobacter hansenii, Agromyces glycinophilus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp. or a combination thereof.

41. The composition according to any one of claims 29 or 31 - 40, wherein the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrabacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof.

42. The composition according to any one of claims 29 or 31 - 41, wherein the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrabacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof.

43. The composition according to any one of claims 29 or 31 - 42, wherein the one or more microbial strains are or comprise Bacillus subtilis.

44. The composition according to any one of claims 29 or 31 - 43, wherein the composition comprises two or more microbial strains.

45. The composition according to any one of claims 29 or 31 - 44, wherein the composition comprises five or more microbial strains.

46. The composition according to any one of claims 29 or 31 - 45, wherein the composition comprises ten or more microbial strains.

47. The composition according to any one of claims 29 - 46, wherein the composition is for topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal, ocular, intravitreal, or suprachoroidal administration.

48. The composition according to claim 47, wherein the composition is for oral administration.

49. The composition according to claim 47, wherein the composition is for intravenous administration.

50. The composition according to any one of claims 29 - 49, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection, or eye drop.

51. The composition according to any one of claims 29 or 31 - 50, wherein each of the one or more microbial strains is present in the composition at a concentration of 10 1 to 10 15 CFU.

52. The composition according to any one of claims 29 or 31 - 50, wherein each of the one or more microbial strains is present in the composition at a concentration of at least 10 6 CFU.

53. Use of the composition according to any one of claims 29 - 52 for modulating one or more microbial metabolites in a subject.

54. Use of the composition according to any one of claims 29 - 52 for modulating one or more characteristics in a subject.

55. Use of the composition according to claim 54, wherein the one or more characteristics are or comprise: (i) the level of cell viability; (ii) the level or activity of nucleic acids or proteins, or their forms; (iii) weight gain; (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride level; (vii) blood cholesterol level; (viii) oxidative stress; or (ix) inflammation.

56. Use of the composition according to any one of claims 29 - 52 for characterizing the ability of one or more microbial strains to modulate one or more microbial metabolites in a subject.

57. Use of the composition according to any one of claims 29 - 52 for treating or ameliorating a disease, disorder, or condition in a subject, wherein the disease, disorder, or condition is an insulin - related disease, disorder, or condition associated with one or more microbial metabolites.

58. The use according to claim 57, wherein the disease, disorder or condition is or comprises diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperglycemic hyperosmolar state, gestational diabetes, diabetic dermopathy, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes or polycystic ovary syndrome (PCOS).

59. The use of the composition according to claim 58, wherein the disease, disorder or condition is diabetes.

60. A method for screening a microbial strain, the method comprising: contacting the microbial strain with a culture comprising pancreatic cells or a pancreatic cell line that mimics an insulin-related disease, disorder or condition, and determining whether the microbial strain has altered a characteristic of the culture, wherein the characteristic is related to the insulin-related disease, disorder or condition.

61. The method according to claim 60, wherein the determining step comprises comparing the characteristic before and after performing the contacting step.

62. The method according to claim 60, wherein the determining step comprises comparing the characteristic after the contacting step with a comparable reference.

63. The method according to claim 62, wherein the comparable reference is a historical reference.

64. The method according to claim 63, wherein the comparable reference is a negative control reference.

65. The method according to claim 63, wherein the comparable reference is a positive control reference.

66. The method according to any one of claims 60-65, wherein the characteristic is the level of cell viability.

67. The method according to any one of claims 60-65, wherein the characteristic is the level or activity of a nucleic acid or protein, or a form thereof.

68. The method according to any one of claims 60-65, wherein the characteristic is or comprises weight gain.

69. The method according to any one of claims 60-65, wherein the characteristic is or comprises fat accumulation in hepatocytes.

70. The method according to any one of claims 60-65, wherein the characteristic is or comprises lipid accumulation in hepatocytes.

71. The method according to any one of claims 60-65, wherein the characteristic is or comprises triglyceride level.

72. The method according to any one of claims 60-65, wherein the characteristic is or comprises cholesterol level.

73. The method according to any one of claims 60-65, wherein the characteristic is or comprises inflammation.

74. The method according to any one of claims 60-65, wherein the microbial strain has altered one or more characteristics of the culture, wherein the one or more characteristics are related to the insulin-related disease, disorder or condition, and wherein the one or more characteristics are or comprise (i) the level of cell viability; (ii) the level or activity of a nucleic acid or protein, or a form thereof; (iii) weight gain; (iv) fat accumulation in the liver; (v) Lipid accumulation in the liver; (vi) Blood triglyceride levels; (vii) Blood cholesterol levels; (viii) Oxidative stress; or (ix) Inflammation.

75. A method comprising: Administering to a subject a composition comprising one or more microbial strains or microbial components.

76. A method comprising: Administering to a subject a composition comprising one or more microbial metabolites.

77. The method according to claim 75 or 76, wherein the subject is a mammal.

78. The method according to any one of claims 75-77, wherein the subject is a human.

79. The method according to claim 75 or any one of claims 77-78, wherein the one or more microbial strains are from a mammalian microbiome.

80. The method according to claim 75 or any one of claims 77-79, wherein the one or more microbial strains are from a human microbiome.

81. The method according to claim 80, wherein the human microbiome is the microbiome of the subject.

82. The method according to claim 81, wherein the human microbiome is administered to maintain or regulate the microbiome of the subject.

83. The method according to any one of claims 75-82, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3 or Appendix 4.

84. The method according to any one of claims 75-83, wherein the one or more microbial metabolites are or comprise bile acids.

85. The method according to any one of claims 75-84, wherein the one or more microbial metabolites are or comprise tauroursodeoxycholic acid.

86. The method according to any one of claims 75-83, wherein the one or more microbial components or microbial metabolites are butyrylcarnitine, theobromine, 4-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro) or any combination thereof.

87. The method according to any one of claims 75 - 83, wherein the one or more microbial components or microbial metabolites are 4 - hydroxyphenylpyruvic acid, ectoine, gramine, N - acetyl - L - phenylalanine, Nε - acetyl - L - lysine, stachydrine, trigonelline, 3 - ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG - methyl - L - arginine, trans - urocanic acid, N - acetyl - L - leucine, sarcosine, isobutyryl carnitine, b - hydroxyisovaleric acid, L - theanine / N5 - ethylglutamine, 5 - hydroxylysine, phenylacetylglycine, betaine, hydroxyproline, picolinic acid, 2 - aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3 - phosphate, argininosuccinic acid, creatine, terephthalic acid, homoarginine, mucic acid, homocysteine sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3 - indolyl sulfate, nicotinamide, N - formylglycine, ureidoethanolate, N - methylproline, glucaric acid, butyryl carnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenylacetylglycine, diethanolamine, phosphocholine, guanidinosuccinic acid, N - acetylhistidine, glyceric acid, S - methylmethionine, cysteine glutathione disulfide, kynurenine, N - acetylphenylalanine, threonic acid, malic acid, 7,8 - dihydrobiopterin, homovanillic acid, taurocholic acid, 5 - methoxyindoleacetic acid, butyrate, b - hydroxyisovaleric acid, 2 - oxoglutaric acid, N - acetyltryptophan, thiazolidine - 4 - carboxylic acid, hypotaurine, cholalic acid, acetoacetic acid, ethanolamine, guanidinoacetic acid, S - sulfocysteine, myristic acid C14:0XA0027 or any combination thereof.

88. The method according to any one of claims 75 or 77 - 87, wherein the one or more microbial strains are or include Acetobacter gluconicum Hansen, Agrobacterium tumefaciens glycolicum, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp. or a combination thereof.

89. The method according to any one of claims 75 or 77 - 88, wherein the one or more microbial strains are or include Acetobacter gluconicum Hansen, Agrobacterium tumefaciens glycolicum, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp. or a combination thereof.

90. The method according to any one of claims 75 or 77 - 89, wherein the one or more microbial strains are or include Acetobacter gluconicum Hansen, Agrobacterium tumefaciens glycolicum, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve or a combination thereof.

91. The method according to any one of claims 75 or 77 - 90, wherein the one or more microbial strains are or include Bacillus subtilis.

92. The method according to any one of claims 75 or 77 - 91, wherein the composition comprises two or more microbial strains.

93. The method according to any one of claims 75 or 77 - 92, wherein the composition comprises five or more microbial strains.

94. The method according to any one of claims 75 or 77 - 93, wherein the composition comprises ten or more microbial strains.

95. The method according to any one of claims 75 - 94, wherein the composition is administered topically, orally, subcutaneously, intravenously, intramuscularly, intracranially, intrathecally, rectally, ophthalmically, intravitreally or suprachoroidally.

96. The method according to claim 95, wherein the composition is administered orally.

97. The method according to claim 95, wherein the composition is administered intravenously.

98. The method according to any one of claims 75 - 97, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection or eye drop.

99. The method according to any one of claims 75 or 77 - 98, wherein each of the one or more microbial strains is present in the composition at a concentration of 10 1 to 10 15 CFU.

100. The method according to any one of claims 75 or 77 - 99, wherein each of the one or more microbial strains is present in the composition at a concentration of at least 10 6 CFU.

101. The method according to any one of claims 75 - 100, wherein the microbial strain or the microbial metabolite alters a characteristic of the subject.

102. The method according to claim 101, wherein the characteristic is the level of cell viability.

103. The method according to claim 101, wherein the characteristic is the level or activity of a nucleic acid or protein, or its form.

104. The method according to claim 101, wherein the characteristic is or includes weight gain.

105. The method according to claim 101, wherein the characteristic is or includes fat accumulation in the liver.

106. The method according to claim 101, wherein the characteristic is or includes lipid accumulation in the liver.

107. The method according to claim 101, wherein the characteristic is or includes triglyceride level.

108. The method according to claim 101, wherein the characteristic is or includes cholesterol level.

109. The method according to claim 101, wherein the characteristic is or includes inflammation.

110. The method according to claim 101, wherein the microbial strain alters one or more characteristics of the subject, and wherein the one or more characteristics are or include (i) the level of cell viability; (ii) the level or activity of a nucleic acid or protein, or its form; (iii) weight gain; (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride level; (vii) blood cholesterol level; (viii) oxidative stress; or (ix) inflammation.

111. The method according to any one of claims 101 - 110, wherein the characteristic is associated with the insulin - related disease, disorder or condition.

112. A method of characterizing a microbial strain, the method comprising: adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that mimic an insulin - related disease, disorder or condition, and Determine whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder or condition.

113. A method of manufacturing a pharmaceutical treatment, the method comprising characterizing one or more microbial strains, microbial components or microbial metabolites, the method comprising the steps of: adding the one or more microbial strains to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that mimic an insulin-related disease, disorder or condition, and determine whether the one or more microbial strains affect the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder or condition.

114. A method of manufacturing a pharmaceutical treatment, the method comprising: formulating one or more microbial strains or microbial communities in syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection or eye drop.

115. A method of manufacturing a pharmaceutical treatment, the method comprising: formulating one or more microbial metabolites in syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection or eye drop.

116. A method of assessing the ability of a microbial strain to affect one or more characteristics of a culture, the method comprising: adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that mimic an insulin-related disease, disorder or condition, and determine whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder or condition.

117. The method according to any one of claims 112-114 or 116, the method further comprising: measuring the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture before adding the microbial strain, measuring the level of the same one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture after adding the microbial strain, and comparing the level of one or more characteristics measured before adding the microbial strain with the level of one or more characteristics measured after adding the microbial strain.

118. The method according to any one of claims 112-117, wherein the one or more characteristics include: (i) the level of cell viability; (ii) the level or activity of nucleic acid or protein, or its form; (iii) weight gain; (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride level; (vii) blood cholesterol level; (viii) oxidative stress; (ix) inflammation; or (x) a combination thereof.

119. The method according to any one of claims 60-118, wherein the disease, disorder or condition is or comprises diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic endoneuritic rash, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes or polycystic ovary syndrome (PCOS).

120. A composition for treating or preventing an insulin-related disease, disorder or condition, the composition comprising one or more microbial strains or microbial components.

121. A composition for treating or preventing an insulin-related disease, disorder or condition, the composition comprising one or more microbial metabolites.

122. The composition used as claimed in claim 120, wherein the one or more microbial strains are from the mammalian microbiome.

123. The composition used as claimed in claim 120 or 122, wherein the one or more microbial strains are from the human microbiome.

124. The composition used as claimed in claim 123, wherein the human microbiome is the microbiome of the subject.

125. The composition used as claimed in claim 124, wherein the human microbiome is administered to maintain or modulate the microbiome of the subject.

126. The composition used as claimed in any one of claims 120-125, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3 or Appendix 4.

127. The composition used as claimed in any one of claims 120-126, wherein the one or more microbial metabolites are or comprise bile acids.

128. The composition used as claimed in any one of claims 120-127, wherein the one or more microbial metabolites are or comprise tauroursodeoxycholic acid.

129. The composition used as claimed in any one of claims 120-126, wherein the one or more microbial components or microbial metabolites are butyrobetaine, theobromine, 4-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro) or any combination thereof.

130. The composition used as described in any one of claims 120 - 126, wherein one or more microbial components or microbial metabolites are 4 - hydroxyphenylpyruvic acid, ectoine, gramine, N - acetyl - L - phenylalanine, Nε - acetyl - L - lysine, stachydrine, trigonelline, 3 - ureidopropionic acid, theobromine, hippuric acid, imidopropionic acid, NG - methyl - L - arginine, trans - urocanic acid, N - acetyl - L - leucine, sarcosine, isobutyryl carnitine, b - hydroxyisovaleric acid, L - theanine / N5 - ethylglutamine, 5 - hydroxylysine, phenylacetylglycine, betaine, hydroxyproline, picolinic acid, 2 - aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3 - phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013C6H6O4S, 3 - indolyl sulfate, nicotinamide, N - formylglycine, ureidoethanolate, N - methylproline, glucaric acid, butyryl carnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenylacetylglycine, diethanolamine, phosphocholine, guanidinosuccinic acid, N - acetylhistidine, glyceric acid, S - methylmethionine, cysteine glutathione disulfide, kynurenine, N - acetylphenylalanine, threonic acid, malic acid, 7,8 - dihydrobiopterin, homovanillic acid, taurocholic acid, 5 - methoxyindoleacetic acid, butyrate, b - hydroxyisovaleric acid, 2 - oxoglutaric acid, N - acetyltryptophan, thioproline, hypotaurine, cholalic acid, acetoacetic acid, ethanolamine, guanidinoacetic acid, S - sulfocysteine, myristic acid C14:0XA0027 or any combination thereof.

131. The composition used as described in any one of claims 120 or 122 - 130, wherein the one or more microbial strains are or include Acetobacter gluconicum hansenii, Agrobacterium tumefaciens glycolicum, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp. or a combination thereof.

132. The composition used as described in any one of claims 120 or 122 - 131, wherein the one or more microbial strains are or include Acetobacter gluconicum hansenii, Agrobacterium tumefaciens glycolicum, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp. or a combination thereof.

133. The composition used as described in any one of claims 120 or 122 - 132, wherein the one or more microbial strains are or include Acetobacter gluconicum hansenii, Agrobacterium tumefaciens glycolicum, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve or a combination thereof.

134. The composition used as described in any one of claims 120 or 122 - 133, wherein the one or more microbial strains are or include Bacillus subtilis.

135. The composition used as described in any one of claims 120 or 122 - 134, wherein the composition comprises two or more microbial strains.

136. The composition used as described in any one of claims 120 or 122 - 135, wherein the composition comprises five or more microbial strains.

137. The composition used as described in any one of claims 120 or 122 - 136, wherein the composition comprises ten or more microbial strains.

138. The composition used as described in any one of claims 120 - 137, wherein the composition is for topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal, ocular, intravitreal or suprachoroidal administration.

139. The composition used as described in claim 138, wherein the composition is for oral administration.

140. The composition used as described in claim 138, wherein the composition is for intravenous administration.

141. The composition used as described in any one of claims 120 - 140, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injection or eye drop.

142. The composition used as described in any one of claims 120 or 122 - 141, wherein each of the one or more microbial strains is present in the composition at a concentration of 10 1 to 10 15 CFU.

143. The composition used as described in any one of claims 120 or 122 - 141, wherein each of the one or more microbial strains is present in the composition at a concentration of at least 10 6 CFU.

144. The composition used as described in any one of claims 120 - 143, wherein the insulin - related disease, disorder or condition is or includes diabetes, obesity, cardiovascular disease, non - alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic endoneuritic rash, diabetic neuropathy, diabetic foot ulcer, maturity - onset diabetes of the young, pancreatogenic diabetes or polycystic ovary syndrome (PCOS).

145. The composition used as described in claim 144, wherein the insulin - related disease, disorder or condition is diabetes.

146. An injection comprising the composition as described in any one of claims 29 - 52.

147. A food supplement comprising the composition as described in any one of claims 29 - 52.

148. A kit comprising the composition as described in any one of claims 29 - 52 or 120, the kit being for treating or preventing an insulin - related disease, disorder or condition.

149. The kit as described in claim 148, which comprises a monitoring device.

150. The kit as described in claim 149, wherein the monitoring device is a blood glucose monitor.

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