Compositions containing indole-derived metabolites and methods of use thereof

Abstract

Some aspects of the disclosure relate to compositions comprising fermentates of bacterial strains capable of producing indole-containing compounds such as ILA and lAld. Some aspects relate to nutritional supplements, and food products containing such fermentates, and methods of use thereof.

Description

[0001] PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0002] COMPOSITIONS CONTAINING INDOLE-DERIVED METABOLITES AND METHODS OF USE THEREOF RELATED APPLICATIONS

[0003] This application claims the benefit under 35 U.S.C. § 119(e) of the earlier filing dates of U.S. Provisional Application No. 63 / 557,070, filed February 23, 2024, and U.S. Provisional Application No. 63 / 738,662, filed December 24, 2024, the contents of each of which are incorporated by reference herein in their entirety.

[0004] GOVERNMENT LICENSE RIGHTS

[0005] This invention was made with government support under Award # 2054208 from the National Science Foundation (NSF). The government has certain rights in the invention.

[0006] REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0007] The contents of the electronic sequence listing (I055870006WO00-SEQ-NTJ.xml; Size: 24,928,554 bytes; and Date of Creation: February 20, 2025) are herein incorporated by reference in their entirety.

[0008] BACKGROUND

[0009] Ingestion of safe bacteria with the intention of benefiting one’s health is a centuries old practice that has seen a surge of interest in the early 21st century due to a better mechanistic understanding of the influence of the microbiome on organismal health. Application of these beneficial microbes, such as probiotics and postbiotics, is now being used in companion animals, namely canines, with the aim to modulate immune responses to combat undesirable symptoms e.g., itching, diarrhea, vomiting.

[0010] SUMMARY

[0011] Aspects of the disclosure relate to compositions and methods for reducing inflammation in mammals, such as companion dogs. Provided are compositions comprising bacterial fermentates that include indole-containing metabolites e.g., indole- 3 -lactic acid (ILA) and indole-3-carboxaldehyde (IAld)). Without wishing to be bound by any particular theory, such indole-containing molecules are thought to act as agonists of the aryl hydrocarbon receptor (AhR), with signal transduction following stimulation having immunoregulatory effects and modulating inflammation. The presence of ILA and / or IAld, and optionally other indole-containing compounds that act as AhR agonists, is thus expected to improve homeostasis by stimulating the AhR in a subject. Also without wishing to be PCT / US25 / 16818 21 February 2025 (21.02.2025) bound by another particular theory, ILA is thought to be converted into indole- 3 -propionic acid (IPA) by host microorganisms (e.g., microbes in the gut), with such produced IPA providing antioxidant and anti-inflammatory effects independently of AhR stimulation. Accordingly, the benefits of fermentates and / or compositions comprising fermentates are not limited to those arising from the presence of ILA and lAld alone.

[0012] It was discovered, surprisingly, that while a bacterial strain may be capable of producing multiple indole-containing compounds from tryptophan as a substrate, the extent of ILA and lAld production varies with fermentation conditions, such that preferential production of ILA or lAld can be achieved by varying conditions in separate fermentations. Glucose concentration was found to affect the production of ILA, such that ILA abundance in a fermentate could be increased by increasing glucose concentration. Production of lAld, however, was influenced by the abundance of nitrogen sources (e.g., yeast extract and peptones), such that production of higher amounts of lAld could be achieved by increasing the amount of nitrogen in a fermentation medium. And, while lAld could also be increased by reducing glucose concentrations (e.g., to levels below 4 g / L), low glucose concentrations limit the availability of carbon for bacterial metabolism. Providing glucose in intermediate concentrations, together with nitrogen, however, allowed for robust lAld production with reduced abundance of ILA, compared to a separate fermentation in which high glucose concentrations are used to promote ILA production.

[0013] It was thus determined that separate fermentations under distinct conditions allows for separately (i) preferential and high production of ILA, and (ii) preferential and high production of lAld, allowing for the production of postbiotic compositions containing high amounts of both ILA and lAld by separate fermentations to produce high amounts of each, and then combining the fermentates. Additionally, producing separate fermentates under separate conditions allows for the production of fermentate mixtures with desired ratios of ILA and lAld for formulation in a composition (e.g., nutritional supplement or food product) for administration with desired doses of ILA and lAld. In contrast to previous fermentation of a bacterium in a single batch, where conditions favorable to production of one metabolite may hinder the other and vice versa, the preferential production of different compounds in separate fermentations allows for the production of postbiotic compositions comprising high amounts of multiple beneficial compounds, such as ILA and lAld, as well as production of postbiotic compositions comprising desired ratios of ILA and lAld.

[0014] Thus, in some aspects, provided are methods of preparing a composition containing high concentrations of both ILA and lAld, by separately fermenting a single bacterial strain PCT / US25 / 16818 21 February 2025 (21.02.2025) under different conditions, such that separate fermentates containing high concentrations of different metabolites are produced and may be combined to produce a composition with improved AhR agonist activity. Pediococcus acidilactici is one such bacterium that produces varying amounts of ILA and lAld under different fermentation conditions. For example, supplementation of fermentation medium with 4-15 g / L glucose, and inclusion of polysorbate 80, vitamin B12, and manganese improved ILA concentration, whereas lAld concentration was highest when polysorbate 80, vitamin B12, and manganese were omitted, and fermentation media contained less than 4 g / L glucose. As another example, Lactiplantibacillus plantarum, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus reuteri, and Lactococcus lactis are capable of ILA production. Additionally, other bacterial strains belonging to species Bacillus subtilis, Weizmannia coagulans, Bacillus pumilus, Bacillus licheniformis, and Bacillus velezensis produce other indole-containing compounds, such as indole-3-acetic acid (IAA).

[0015] In addition to the benefits of AhR agonist activity from indoles, e.g., ILA and lAld, fermentates may be produced using bacterial strains that produce low levels of undesired indole-containing compounds or do not produce such undesired compounds at all. In comparison to use of tryptophan itself, which may be converted into a number of compounds by gut microbiota, the use of fermentates comprising specified compounds but that lack or have only low levels of undesired compounds allows such fermentate-containing compositions to avoid unwanted side effects of tryptophan metabolism into such undesired compounds (e.g., indoxyl sulfate).

[0016] Accordingly, some aspects of the disclosure relate to a postbiotic composition comprising a first fermentate and a second fermentate of a bacterial strain, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 (Pediococcus acidilactici) or SEQ ID NO: 7 (Lactiplantibacillus plantarum), and wherein the first fermentate comprises indole- 3 -lactic acid (ILA) and the second fermentate comprises indole-3-carboxaldehyde (lAld). In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the bacterial strain belongs to the species Pediococcus acidilactici. In some embodiments, the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8. In some embodiments, the bacterial strain comprises a genome with at least 95.0% average nucleotide identity and an alignment fraction of at least PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0017] 50.0% to the reference genome of SEQ ID NO: 8. In some embodiments, the first fermentate is produced by incubating the bacterial strain under aerobic or microaerobic conditions in a fermentation medium comprising: 1 g / L to 12 g / L tryptophan; 4 g / L to 15 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12. In some embodiments, the second fermentate is produced by incubating the bacterial strain under the following conditions: 1 g / L to 12 g / L tryptophan; less than 4 g / L glucose; and no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12. In some embodiments, a bacterial strain belonging to the species Clostridium sporogenes produces indole-3-propionic acid (IPA) in the presence of the postbiotic composition.

[0018] Some aspects relate to a postbiotic composition comprising two or more fermentates of one or more bacterial strains, wherein a first fermentate comprises indole-3-lactic acid (ILA), and a second fermentate comprises indole-3-carboxaldehyde (lAld).

[0019] In some embodiments, the postbiotic composition further comprises indole- 3 -pyruvic acid (IPyA), indole- 3 -propionic acid (IPA), 3-indoleacrylic acid (IA), and / or indole-3-acetic acid (IAA).

[0020] In some embodiments, the first fermentate and / or the second fermentate further comprises indole-3-pyruvic acid (IPyA) and / or indole-3-propionic acid (IPA). In some embodiments, the postbiotic composition does not comprise indole-3-acetic acid (IAA).

[0021] In some embodiments, the postbiotic composition further comprises one or more additional fermentates of one or more additional bacterial strains selected from the group consisting of Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans, and Lactiplantibacillus plantarum. In some embodiments, the one or more additional fermentates comprise ILA. In some embodiments, the one or more additional fermentates comprise lAld. In some embodiments, the one or more additional fermentates comprise IPyA and / or IPA. In some embodiments, the postbiotic composition comprises at least 0.1% (w / w) ILA. In some embodiments, the postbiotic composition comprises at least 0.015% (w / w) lAld.

[0022] In some embodiments, the postbiotic composition further comprises IPyA and / or IPA. In some embodiments, the postbiotic composition comprises at least 10% w / w IPyA relative to ILA. In some embodiments, the postbiotic composition comprises at least 1% w / w IPA relative to lAld. In some embodiments, the postbiotic composition comprises at least 10% w / w IAA relative to ILA.

[0023] In some embodiments, the postbiotic composition comprises less than 200% w / w tryptophan relative to ILA. In some embodiments, the postbiotic composition does not PCT / US25 / 16818 21 February 2025 (21.02.2025) comprise indoxyl sulfate. In some embodiments, the postbiotic composition comprises less than 10% w / w indole-3-acetic acid (IAA) relative to ILA. In some embodiments, the postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA. In some embodiments, the postbiotic composition comprises less than 0.5% w / w indole-3-acetamide relative to ILA.

[0024] In some embodiments, each of the fermentates is in powdered form. In some embodiments, each of the fermentates is lyophilized or spray-dried.

[0025] In some embodiments, each of the fermentates is present in an amount of 0.1 to 1000 mg. In some embodiments, the postbiotic composition comprises a total amount of fermentates of 0.2 to 2000 mg.

[0026] In some embodiments, the postbiotic composition further comprises an excipient. In some embodiments, the excipient is a tapioca-based maltodextrin.

[0027] In some embodiments, the postbiotic composition is in a form for topical administration. In some embodiments, the postbiotic composition is in a form for oral administration. In some embodiments, the postbiotic composition is in the form of a dry flowable powder.

[0028] In some embodiments, the postbiotic composition reduces inflammation in a subject. In some embodiments, the postbiotic composition treats or prevents a symptom of inflammation in a subject. In some embodiments, the postbiotic composition treats or prevents diarrhea in a subject. In some embodiments, the postbiotic composition treats or prevents vomiting in a subject. In some embodiments, the postbiotic composition treats or prevents itching in a subject. In some embodiments, the postbiotic composition treats or prevents a symptom of allergy in a subject.

[0029] In some embodiments, the subject is a carnivore. In some embodiments, the subject is a mammal. In some embodiments, the subject is a domesticated animal. In some embodiments, the subject is a dog, cat, rabbit, guinea pig, hamster, or ferret. In some embodiments, the subject is a dog.

[0030] Some aspects relate to a nutritional supplement or food product comprising a postbiotic composition described herein. In some embodiments, the nutritional supplement or food product comprises a meat or animal-derived material. In some embodiments, the meat or animal-derived material is beef, chicken, eggs, turkey, lamb, fish, blood marrow, and / or bone marrow. In some embodiments, the nutritional supplement or food product comprises a grain. In some embodiments, the grain is wheat, corn, rice, oats, and / or barley. In some embodiments, the nutritional supplement or food product comprises a fiber. In some PCT / US25 / 16818 21 February 2025 (21.02.2025) embodiments, the fiber is sugar beet pulp, chicory pulp, chicory, coconut endosperm fiber, wheat fiber, fructooligosaccharides, galactooligosaccharides, and / or inulin.

[0031] In some embodiments, the nutritional supplement or food product comprises an algae. In some embodiments, the algae is Ascophyllum nodosum, Spirulina, and / or Fucus vesiculosus.

[0032] In some embodiments, the nutritional supplement or food product comprises a gelatinized starch matrix.

[0033] Some aspects relate to a topical preparation comprising a postbiotic composition as described herein. Form factors of compositions suitable for administration include, without limitation, shampoos (liquid cleansing preparations), creams (semi-solid, water-based emulsions), ointments (viscous, greasy preparations with an oil base), sprays (liquids formulated in aerosol or pump form), spot-on solutions (liquid preparations formulated for application to a single area of a surface), lotions (semi-solid, water-based emulsions less viscous than creams), gels (semi-solid preparations, which are typically clear), foams (aerated preparations that are dispensed as mousse), balms (waxy preparations for coating a surface), and powders (dry preparations for dusting a surface). In some embodiments, a postbiotic composition is in the form of a shampoo. In some embodiments, a postbiotic composition is in the form of a cream. In some embodiments, a postbiotic composition is in the form of an ointment. In some embodiments, a postbiotic composition is in the form of a spray. In some embodiments, a postbiotic composition is in the form of a spot-on solution. In some embodiments, a postbiotic composition is in the form of a lotion. In some embodiments, a postbiotic composition is in the form of a gel. In some embodiments, a postbiotic composition is in the form of a foam. In some embodiments, a postbiotic composition is in the form of a balm. In some embodiments, a postbiotic composition is in the form of a powder.

[0034] Some aspects relate to a method comprising administering a postbiotic composition, nutritional supplement or food product as described herein, to a subject in need thereof. In some embodiments, the administration is oral administration. In some embodiments, the administration is topical administration.

[0035] In some embodiments, the administering: (i) reduces inflammation in the subject; (ii) treats or prevents a symptom of inflammation in the subject; (iii) treats or prevents diarrhea in the subject; (iv) treats or prevents vomiting in the subject; (v) treats or prevents itching in the subject; and / or (vi) treats or prevents a symptom of allergy in the subject. In some embodiments, the administering: (i) treats or prevents skin inflammation, skin irritation, PCT / US25 / 16818 21 February 2025 (21.02.2025) and / or skin damage in the subject; (ii) treats or prevents gut inflammation or gastrointestinal distress in the subject; (iii) treats or prevents neuroinflammation in the subject; (iv) treats or prevents weight-related inflammation in the subject; (v) limits weight gain in the subject; (vi) promotes weight loss in the subject; (vii) treats or prevents overweight status or obesity in the subject; (viii) treats or prevents anxiety in the subject; (ix) treats or prevents seasonal allergy in the subject; (x) treats or prevents food allergy in the subject; and / or (xi) improves coat quality in the subject.

[0036] In some embodiments, the subject is a carnivore. In some embodiments, the subject is a mammal. In some embodiments, the subject is a domesticated animal. In some embodiments, the subject is a dog, cat, rabbit, guinea pig, hamster, or ferret. In some embodiments, the subject is a dog. In some embodiments, the subject is a cat.

[0037] Some aspects relate to a method for producing a postbiotic composition, the method comprising: (i) fermenting a first bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 (Pediococcus acidlaclici) or SEQ ID NO: 7 (Lactiplantibacillus plantarum) in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA; (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld; and (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the bacterial strain belongs to the species Pediococcus acidilactici. In some embodiments, the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8. In some embodiments, the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 50.0% to the reference genome of SEQ ID NO: 8. In some embodiments, the first fermentate is produced by incubating the bacterial strain under aerobic or microaerobic conditions in a fermentation medium comprising: 1 g / L to 12 g / L tryptophan; 4 g / L to 15 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12. In some embodiments, the second fermentate is produced by incubating the bacterial strain under the following conditions: 1 g / L to 12 g / L tryptophan; less than 4 g / L glucose; and no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12. In some embodiments, a bacterial strain belonging to the species Clostridium PCT / US25 / 16818 21 February 2025 (21.02.2025) sporogenes produces indole- 3 -propionic acid (IPA) in the presence of the postbiotic composition.

[0038] In some embodiments, the first and / or second fermentate further comprises indole-3- pyruvic acid (IPyA) and / or indole- 3 -propionic acid (IPA). In some embodiments, the first and / or second fermentate further comprises 3-indoleacrylic acid (IA), and / or indole-3-acetic acid (IAA).

[0039] In some embodiments, the method further comprises fermenting one or more additional bacterial strains to produce one or more additional fermentates, and combining the one or more additional fermentates with the first and second fermentates or the postbiotic composition, wherein the one or more additional bacterial strains are selected from the group consisting of Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans.

[0040] In some embodiments, the one or more additional fermentates comprise ILA and / or lAld. In some embodiments, the one or more additional fermentates comprise IPyA and / or IPA. In some embodiments, the postbiotic composition comprises at least 0.1% (w / w) ILA. In some embodiments, the postbiotic composition comprises at least 0.015% (w / w) lAld.

[0041] In some embodiments, the postbiotic composition further comprises IPyA and / or IPA. In some embodiments, the postbiotic composition comprises at least 10% w / w IPyA relative to ILA. In some embodiments, the postbiotic composition comprises at least 1% w / w IPA relative to lAld.

[0042] In some embodiments, the postbiotic composition comprises less than 200% w / w tryptophan relative to ILA. In some embodiments, the postbiotic composition does not comprise indole. In some embodiments, the postbiotic composition does not comprise indoxyl sulfate. In some embodiments, the postbiotic composition comprises less than 10% w / w indole-3-acetic acid (IAA) relative to ILA. In some embodiments, the postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA. In some embodiments, the postbiotic composition comprises less than 0.5% w / w indole-3-acetamide relative to ILA.

[0043] In some embodiments, the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising: 1 g / L to 50 g / L tryptophan; 100 g / L to 200 g / L glucose; manganese; and 10 g / L to 125 g / L of one or more nitrogen sources. In some embodiments, the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising: 1 g / L to 50 g / L tryptophan; 1 g / L to 100 g / L glucose; manganese; and 125 g / L to 200 g / L of one or more PCT / US25 / 16818 21 February 2025 (21.02.2025) nitrogen sources. In some embodiments, the one or more nitrogen sources comprise yeast extract or peptones.

[0044] In some embodiments: the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin Bl 2; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B 12. In some embodiments: the first fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, less than 4 g / L glucose, no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

[0045] In some embodiments, the second fermentate comprises more lAld than the first fermentate. In some embodiments, the first fermentate comprises more ILA than the second fermentate.

[0046] In some embodiments, the first fermentation medium and / or the second fermentation medium comprises tryptophan at a concentration of 1 to 12 g / L.

[0047] In some embodiments, the first fermentation medium comprises glucose at a concentration of at least 4 g / L. In some embodiments, the first fermentation medium or the comprises glucose at a concentration of at least 4 to 15 g / L. In some embodiments, the second fermentation medium comprises glucose at a concentration of less than 4 g / L.

[0048] In some embodiments, the first fermentation medium comprises polysorbate 80, manganese, and / or vitamin B12. In some embodiments, the second fermentation medium does not comprise polysorbate 80, does not comprise manganese, and does not comprise vitamin B12.

[0049] In some embodiments, the fermenting of (i) and / or the fermenting of (ii) is for 2 to 7 days. In some embodiments, the fermenting of (i) and / or the fermenting of (ii) is under aerobic or microaerobic conditions.

[0050] In some embodiments, the method further comprises separating bacterial cells from the first fermentate and / or second fermentate before combining the first and second fermentates. In some embodiments, the method further comprises heat-killing bacterial strains in the first fermentate and / or second fermentate before combining the first and second PCT / US25 / 16818 21 February 2025 (21.02.2025) fermentates. In some embodiments, the method further comprises concentrating the first fermentate and / or second fermentate before combining the first and second fermentates. In some embodiments, the method further comprises drying the first fermentate and / or second fermentate before combining the first and second fermentates.

[0051] Some aspects relate to a postbiotic composition produced by a method described herein.

[0052] BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The accompanying drawings, which constitute a part of this specification, illustrate several embodiments of the disclosure and together with the description, serve to explain the principles of the disclosure.

[0054] FIG. 1 shows relative indole-3-lactic acid (ILA) and indole-3-carboxaldehyde (lAld) titers obtained during fermentation with Pediococcus acidilactici at different glucose concentrations.

[0055] FIGs. 2A and 2B show ILA (FIG. 2A) and lAld (FIG. 2B) titers obtained during fermentation with Pediococcus acidilactici at exemplary glucose concentrations for each compound, respectively.

[0056] FIGs. 3A and 3B show relative ILA (FIG. 3A) and lAld (FIG. 3B) titers obtained during fermentation with Pediococcus acidilactici with or without supplementation of media with polysorbate 80 (1 g / L final concentration), manganese (0.1 g / L final concentration), and vitamin B12 (0.001 mg / L final concentration).

[0057] FIGs. 4A and 4B show relative ILA (FIG. 4A) and lAld (FIG. 4B) titers obtained during fermentation with Pediococcus acidilactici in different oxygenation conditions.

[0058] FIG. 5 shows AhR agonism of ILA and lAld in human (left column of each pair) and canine (right column of each pair) cell lines. Data are normalized to the positive control (methylcholanthrene) .

[0059] FIG. 6 shows canine AhR activation by Strain 1 Postbiotic Fermentate Mixture (an exemplary postbiotic composition) or a mixture of ILA and lAld.

[0060] FIGs. 7A and 7B show reduction in IFN-y (FIG. 7A) and TNF-a (FIG. 7B) from PBMCs exposed to phytohemagglutinin (PHA, - control), Strain 1 Postbiotic Fermentate Mixture, a mixture of purified ILA and lAld, or inhibitor dexamethasone (+ control).

[0061] FIGs. 8A-9B show results of a double-blind, multi-group controlled study as described in Example 4, which was conducted to evaluate the efficacy of the postbiotic of Example 2, containing the first and second fermentates of Strain 1 in reducing canine itching. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0062] FIG. 8A shows change in seconds scratching per day, by group, as measured by activity monitor. Lines and error bars indicate the mean and standard deviation. Points represent individual participant data. FIG. 8B shows change in PVAS score, by group. Lines and error bars indicate the mean and standard deviation. Points represent individual participant data. FIG. 8C shows a summary of changes to coat quality. Both groups started with a median score of 1 (mean = 1.33), indicating a typical quality coat. FIG. 9A shows diversity of gut microbiomes as measured by Shannon index. FIG. 9B shows Taxa with significant log-fold changes in placebo control (top) and postbiotic (bottom) groups.

[0063] FIG. 10 shows production of IPA by Clostridium sporogenes in the presence of Strain 1 Postbiotic Fermentate Mixture or media only (- control).

[0064] FIG. 11 shows activation of canine PXR by Strain 1 Postbiotic Fermentate Mixture, normalized to positive and negative controls.

[0065] FIG. 12A shows activation of human Nrf2 by Strain 1 Postbiotic Fermentate Mixture, normalized to positive and negative controls. FIG. 12B shows a dose-response curve of Nrf2 activation by Strain 1 Postbiotic Fermentate Mixture at a range of concentrations.

[0066] FIG. 13A shows relative concentrations of ILA produced in conditions with varying concentrations of glucose and nitrogen sources (e.g., yeast extract and peptones). FIG. 13B shows relative concentrations of lAld produced in conditions with varying concentrations of glucose and nitrogen sources (e.g., yeast extract and peptones).

[0067] FIG. 14A shows relative concentrations of ILA produced in the presence or absence of manganese. FIG. 14B shows relative concentrations of lAld produced in the presence or absence of manganese.

[0068] DETAILED DESCRIPTION

[0069] Aspects of the disclosure relate to compositions, nutritional supplements, and / or food products comprising fermentates of one or more bacterial strains that contain indolecontaining molecules including indole- 3 -lactic acid (ILA) and indole-3-carboxaldehyde (lAld). Without wishing to be bound by any particular theory, indole-containing molecules produced by metabolism of L-tryptophan, such as ILA and lAld, are thought to act as agonists for the aryl hydrocarbon receptor (AhR). Ligand-binding to AhR facilitates a conformational change exposing the nuclear localization signal, causing translocation to the nucleus, and dimerization of ligand-bound AhR with AHR-nuclear translocator (ARNT) to produce a transcription factor complex that causes chromatin remodeling and transcription of target genes, including 1L6, 1L22, vascular endothelial growth factor A VEGFA), cytochrome PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0070] P450 1A1 (CYP1A1), cytochrome P450 1B1 (CYP1B1), and prostaglandin G / H synthase 2 (PTGS ). See, e.g., Hubbard et al., Drug Metab Dispos. (2015) 43(10):1522-1535. Indolecontaining molecules (e.g., indole-3-carbinol, or indole) have been shown to inhibit proinflammatory responses in vivo and increase barrier function in intestinal epithelial cells. Without wishing to be bound by a particular theory, the compositions described herein comprising fermentates including ILA and lAld are useful for stimulation of AhRs, particularly canine AhR, thereby promoting homeostasis by alleviating inflammation and promoting tissue barrier integrity. Fermentates of other bacterial strains may also comprise ILA or lAld, and / or other indole-containing compounds when appropriately cultured, such as indole-3-pyruvic acid (IpyA), indole-3-propionic acid (IPA), 3-indoleacrylic acid (IA), and indole-3-acetic acid (IAA).

[0071] Embodiments of compositions and methods described herein relate to combinations of ILA and lAld. It was discovered that these compounds act as AhR agonists, in particular for canine AhRs on cultured canine cells. Bacterial strains, such as Pediococcus acidilactici and Lactiplantibacillus plantarum, were able to produce both ILA and lAld. It was also discovered that common fermentation conditions (e.g., adequate tryptophan concentration, similar duration, similar temperature) could be used to produce both compounds, but variations in glucose concentration and supplementation with manganese, polysorbate 80, and vitamin B12 could be used to direct fermentation towards preferential production of one compound or the other. For example, glucose concentrations between 4-15 g / L and inclusion of manganese, polysorbate 80, and vitamin B12 supplementation provided the highest amount of ILA, while glucose concentrations below 4 g / L and lack of manganese, polysorbate 80, and vitamin B12 supplementation provided the highest amounts of lAld. Separate fermentations under different conditions allow for separate preferential and high production of each compound, with combinations of the resulting fermentates providing compositions rich in both ILA and lAld. Such high concentrations of both ILA and lAld is contrasted with that achieved using a single fermentation, as the Examples demonstrate that conditions favorable to production of ILA may hinder production of lAld and vice versa (see Example 1). These and other fermentates of other bacterial strains, such as Bacillus subtilis, Weizmannia coagulans, Bacillus pumilus, Bacillus licheniformis, and Bacillus velezensis, may be used to produce postbiotic compositions comprising ILA and lAld, and optionally other indole-containing compounds such as IPA, IPyA, IA, and IAA.

[0072] Compositions comprising bacteria and / or fermentates thereof may be in any suitable form, such as a dry powder that may be combined with other compositions (e.g., combination PCT / US25 / 16818 21 February 2025 (21.02.2025) with food prior to a meal). Compositions comprising bacteria may also be present as part of another composition, such as a pill, capsule, tablet, chewable matrix, nutritional supplement, and / or food product.

[0073] This disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The disclosure is capable of other embodiments and of being practiced or of being carried out in various ways. Also, the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including,” “comprising,” or “having,” “containing,” “involving,” and variations thereof herein, is meant to encompass the items listed thereafter and equivalents thereof as well as additional items.

[0074] Bacterial strains and fermentates thereof

[0075] Some aspects relate to compositions comprising a bacterial fermentate mixture comprising two or more bacterial fermentates, the first fermentate comprising ILA, and the second fermentate comprising more lAld than the first fermentate. First and second fermentates may be produced by the same bacterial strain, or different bacterial strains.

[0076] Some aspects relate to compositions (and / or nutritional supplements and / or food products comprising a composition) comprising both a first fermentate of a bacterial strain and a second fermentate of the same bacterial strain, where the first fermentate comprises ILA, and the second fermentate comprises lAld.

[0077] Some aspects relate to compositions (and / or nutritional supplements and / or food products comprising a composition) comprising both a first fermentate of a bacterial strain of Pediococcus acidilactici, and a second fermentate of the same bacterial strain of Pediococcus acidilactici, where the first fermentate comprises ILA, and the second fermentate comprises lAld.

[0078] Some aspects relate to compositions (and / or nutritional supplements and / or food products comprising a composition) comprising both a first fermentate of a bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 1, and a second fermentate of the same bacterial strain comprising the same 16S rDNA sequence, where the first fermentate comprises ILA, and the second fermentate comprises lAld. In some embodiments, a bacterial strain, e.g., that produces a fermentate, has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0079] 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 1.

[0080] Some aspects relate to compositions (e.g., postbiotic compositions) (and / or nutritional supplements and / or food products comprising a composition) comprising both a first fermentate of a bacterial strain of Lactiplantibacillus plantarum, and a second fermentate of the same bacterial strain of Lactiplantibacillus plantarum, where the first fermentate comprises ILA, and the second fermentate comprises lAld.

[0081] Some aspects relate to compositions (e.g., postbiotic compositions) (and / or nutritional supplements and / or food products comprising a composition) comprising both a first fermentate of a bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 7, and a second fermentate of the same bacterial strain comprising the same 16S rDNA sequence, where the first fermentate comprises ILA, and the second fermentate comprises lAld. In some embodiments, a bacterial strain, e.g., that produces a fermentate, has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 7.

[0082] Where reference is made to a first fermentate and a second fermentate, the first and second fermentates are to be understood as not being identical in composition, and may contain additional indole-containing compounds beyond those specified. In some embodiments, the first fermentate comprises ILA at a higher amount than lAld. In some embodiments, the second fermentate comprises lAld at a higher amount than ILA. In some embodiments, the first fermentate comprises ILA at a higher amount than lAld, and the second fermentate comprises lAld at a higher amount than ILA.

[0083] In some embodiments, a composition (e.g., postbiotic composition) further comprises one or more additional fermentates of one or more additional bacterial strains. Additional fermentates of additional bacterial strains may comprise, for instance, ILA and / or lAld, thereby contributing to increased ILA and / or lAld content of the composition.

[0084] In some embodiments, an additional fermentate comprises ILA. In some embodiments, an additional fermentate comprises lAld. In some embodiments, an additional fermentate comprises ILA and lAld.

[0085] Additional fermentates of additional bacterial strains may comprise one or more other indole-containing compounds. Such other indole-containing compounds may also contribute PCT / US25 / 16818 21 February 2025 (21.02.2025) to AhR stimulation and its consequent benefits in mitigating inflammation and other symptoms thereof.

[0086] In some embodiments, an additional fermentate comprises IPyA. In some embodiments, an additional fermentate comprises IPA. In some embodiments, an additional fermentate comprises IPyA and IPA.

[0087] In some embodiments, an additional fermentate comprises IA. In some embodiments, an additional fermentate comprises IAA. In some embodiments, an additional fermentate comprises IA and IAA.

[0088] Exemplary bacterial strains capable of producing indole-containing compounds, including ILA, lAld, IPyA, IPA, IA, and / or IAA, are provided below in Table 1. Any bacterial strain corresponding to those of Table 1 (e.g., belonging to the same species, comprising a 16S rDNA sequence with a specified percentage identity to a reference 16S rDNA sequence, and / or comprising a genome with a specified average nucleotide identity (ANI) and alignment fraction (AF) to a reference genome) may be used in production of one or more fermentates (e.g., first fermentate, second fermentate, and / or additional fermentate).

[0089] Table 1: Exemplary bacterial strains capable of producing indole-containing compounds. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0090] In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 1. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 2. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 3. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 4. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 5. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 6. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a PCT / US25 / 16818 21 February 2025 (21.02.2025) bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 7. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 15. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 16. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 17. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 18. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 19. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 20. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 21. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 22.

[0091] In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 3. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 4. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain PCT / US25 / 16818 21 February 2025 (21.02.2025) comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 5. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 6. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 7. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 15. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 16. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 17. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 18. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 19. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 20. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 21. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain comprising at 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 22.

[0092] In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Pediococcus acidilactici. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Bacillus licheniformis . In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Bacillus pumilus. In some embodiments, a composition comprises one or more fermentates (e.g., first, PCT / US25 / 16818 21 February 2025 (21.02.2025) second, and / or additional fermentates) of a bacterial strain belonging to Bacillus subtilis. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Bacillus velezensis. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Weizmannia coagulans. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Lactiplantibacillus plantarum. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Bifidobacterium adolescentis. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Bifidobacterium bifidum. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Bifidobacterium longum. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Lactobacillus acidophilus. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Lactobacillus delbrueckii. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Lactobacillus helveticus. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Lactobacillus reuteri. In some embodiments, a composition comprises one or more fermentates (e.g., first, second, and / or additional fermentates) of a bacterial strain belonging to Lactococcus lactis.

[0093] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 1. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 1 belongs to the species Pediococcus acidilactici. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1 belongs to the species Pediococcus acidilactici. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 1 comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment PCT / US25 / 16818 21 February 2025 (21.02.2025) fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 8. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 8.

[0094] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 2. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 2 belongs to the species Bacillus licheniformis . In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 9. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 9 comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 9. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 9. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 9.

[0095] In some embodiments, a bacterial strain, e.g., that produces a fermentate e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0096] 912%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 3. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 3. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 3 belongs to the species Bacillus pumilus. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 10 comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 10. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%,

[0097] 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%,

[0098] 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%,

[0099] 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 10. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 10.

[0100] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 4. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 4. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 4 belongs to the species Bacillus subtilis. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 11 comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 11. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%,

[0101] 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%,

[0102] 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0103] 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 11. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 11.

[0104] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 5. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 5. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 5 belongs to the species Bacillus velezensis. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 12 comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 12. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 12. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 12.

[0105] In some embodiments, a bacterial strain, e.g., that produces a fermentate e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 6. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 6. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 6 belongs to the species Weiz.inannia coagulans. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 13 comprises a genome with at least 95.0% PCT / US25 / 16818 21 February 2025 (21.02.2025) average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 13. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 13. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 13.

[0106] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 7. In some embodiments, the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 7. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 7 belongs to the species Lactiplantibacillus plantarum. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 14 comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 14. In some embodiments, the bacterial strain comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 14. In some embodiments, the bacterial strain comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 14.

[0107] In some embodiments, a bacterial strain, e.g., that produces a fermentate e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0108] 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 15. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 15 belongs to the species Bifidobacterium adolescentis. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 15 belongs to the species Bifidobacterium adolescentis.

[0109] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 16. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 16 belongs to the species Bifidobacterium bifidum. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 16 belongs to the species Bifidobacterium bifidum.

[0110] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 17. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 17 belongs to the species Bifidobacterium longum. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 17 belongs to the species Bifidobacterium longum.

[0111] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 18. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 18 belongs to the species Lactobacillus acidophilus. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 18 belongs to the species Lactobacillus acidophilus.

[0112] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0113] 912%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 19. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 19 belongs to the species Lactobacillus delbrueckii. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 19 belongs to the species Lactobacillus delbrueckii.

[0114] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 20. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 20 belongs to the species Lactobacillus helveticus. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 20 belongs to the species Lactobacillus helveticus.

[0115] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 21. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 21 belongs to the species Lactobacillus reuteri. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 21 belongs to the species Lactobacillus reuteri.

[0116] In some embodiments, a bacterial strain, e.g., that produces a fermentate (e.g., first, second, and / or additional fermentate), has a 16S rDNA sequence with at least 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% identity to SEQ ID NO: 22. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to SEQ ID NO: 22 belongs to the species Lactococcus lactis. In some embodiments, the bacterial strain comprising a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 22 belongs to the species Lactococcus lactis. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0117] In some embodiments, a bacterial strain that belongs to a particular species has a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to a Species Representative Genome provided in Table 1.

[0118] In some embodiments, a bacterial strain belonging to the species Pediococcus acidilactici comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. CF_000146325.1 or GCF_024970065.1. In some embodiments, the AF is at least 50.0%.

[0119] In some embodiments, a bacterial strain belonging to the species Bacillus licheniformis comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_000011645.1. In some embodiments, the AF is at least 50.0%.

[0120] In some embodiments, a bacterial strain belonging to the species Bacillus pumilus comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_900186955.1, GCA_001938995.1, GCF_002744245.1, GCF_003431975.1, GCF_009937765.1, or GCF_024498355.1. In some embodiments, the AF is at least 50.0%.

[0121] In some embodiments, a bacterial strain belonging to the species Bacillus subtilis comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. CF_000009045.1 or GCF_002153395.1. In some embodiments, the AF is at least 50.0%.

[0122] In some embodiments, a bacterial strain belonging to the species Bacillus velezensis comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_001461825.1. In some embodiments, the AF is at least 50.0%.

[0123] In some embodiments, a bacterial strain belonging to the species Weizmannia coagulans comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_000290615.1. In some embodiments, the AF is at least 50.0%.

[0124] In some embodiments, a bacterial strain belonging to the species Lactiplantibacillus plantarum comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_014131735.1 or GCA_000463075.2. In some embodiments, the AF is at least 50.0%.

[0125] In some embodiments, a bacterial strain belonging to the species Bifidobacterium adolescentis comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_000010425.1. In some embodiments, the AF is at least 50.0%.

[0126] In some embodiments, a bacterial strain belonging to the species Bifidobacterium bifidum comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_001025135.1. In some embodiments, the AF is at least 50.0%. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0127] In some embodiments, a bacterial strain belonging to the species Bifidobacterium longum comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to

[0128] Accession No. GCF_000196555.1 or GCA_905202265.1. In some embodiments, the AF is at least 50.0%.

[0129] In some embodiments, a bacterial strain belonging to the species Lactobacillus acidophilus comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_001591845.1. In some embodiments, the AF is at least 50.0%.

[0130] In some embodiments, a bacterial strain belonging to the species Lactobacillus delbrueckii comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_001433875.1 or GCA_004556255.1. In some embodiments, the AF is at least 50.0%.

[0131] In some embodiments, a bacterial strain belonging to the species Lactobacillus helveticus comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_00160855.1. In some embodiments, the AF is at least 50.0%.

[0132] In some embodiments, a bacterial strain belonging to the species Lactobacillus reuteri comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No.

[0133] GCF_000016825.1, GCF_003072625.1, GCF_013694365.1, GCF_020784725.1,

[0134] GCF_012971035.1, or GCF_020784195.1. In some embodiments, the AF is at least 50.0%.

[0135] In some embodiments, a bacterial strain belonging to the species Lactococcus lactis comprises a genome with at least 95.0% ANI and an AF of at least 20.0% to Accession No. GCF_029023865.1. In some embodiments, the AF is at least 50.0%.

[0136] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 1 (also referred to herein as “Strain 1”) has the highest homology with a bacterial strain of the species Pediococcus acidilactici:

[0137] Strain 1 16S ribosomal RNA coding sequence (16S rDNA)

[0138] AT GAGAGT T T GAT C T T GGC T C AGGAT GAAC GC T GGC GGC GT GC C T AAT AC AT GC AAGT C GAAC GAAC T T C C GT T A ATTGATTATGACGTGCTTGCACTGAATGAGATTTTAACACGAAGTGAGTGGCGGACGGGTGAGTAACACGTGGGT AAC C T GC C C AGAAGC AGGGGAT AAC AC C T GGAAAC AGAT GC T AAT AC C GT AT AAC AGAGAAAAC CGCCTGGTTTT CTTTTAAAAGATGGCTCTGCTATCACTTCTGGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCA C C AAGGC GAT GAT GC GT AGC C GAC C T GAGAGGGT AAT CGGCCACATT GGGAC T GAGAC AC GGC C C AGAC T C C T AC GGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTT TCGGCTCGTAAAGCTCTGTTGTTAAAGAAGAACGTGGGTGAGAGTAACTGTTCACCCAGTGACGGTATTTAACCA GAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCG TAAAGCGAGCGCAGGCGGTCTTTTAAGTCTAATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATTGGAAACTG GGAGACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACC AGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATAC CCTGGTAGTCCATGCCGTAAACGATGATTACTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCA TTAAGTAATCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAAGAATTGACGGGGGCCCGCACAAGCGGTG PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0139] GAGCAT GT GGT T T AAT T C GAAGC T AC GC GAAGAAC CTTACCAGGTCTT GAC AT C T T C T GC C AAC C T AAGAGAT TA GGCGTTCCCTTCGGGGACAGAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAG TCCCGCAACGAGCGCAACCCTTATTACTAGTTGCCAGCATTCAGTTGGGCACTCTAGTGAGACTGCCGGTGACAA ACCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATG GTACAACGAGTTGCGAAACCGCGAGGTTTAGCTAATCTCTTAAAACCATTCTCAGTTCGGACTGTAGGCTGCAAC TCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTAC ACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGCCGGTGGGGTAACCTTTTAGGAGCTAGCCGTCTAA GGTGGGACAGATGATTAGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCGGCTGGATCACCTCCTT (SEQ ID NO : 1 )

[0140] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 2

[0141] (also referred to herein as “Strain 2”) has the highest homology with a bacterial strain of the species Bacillus licheniformis:

[0142] Strain 2 16S ribosomal RNA coding sequence (16S rDNA)

[0143] CACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGA C C T GAGAGGGT GAT C GGC C AC AC T GGGAC T GAGAC AC GGC C C AGAC T C C T AC GGGAGGC AGC AGT AGGGAAT C T T CCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAACTCTGTTGTT AGGGAAGAACAAGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTG CCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTTC T T AAGT C T GAT GT GAAAGC CCCCGGCT C AAC C GGGGAGGGT C AT T GGAAAC T GGGGAAC T T GAGT GC AGAAGAGG AGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGG TCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAAC GATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGT ACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAG C AAC GC GAAGAAC CTTACCAGGTCTT GAC AT C C T C T GAC AAC C C T AGAGAT AGGGC TTCCCCTTC GGGGGC AGAG TGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTT GATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGAC GT C AAAT C AT C AT GC C C C T T AT GAC C T GGGC T AC AC AC GT GC T AC AAT GGGC AGAAC AAAGGGC AGC GAAGC C GC GAGGCTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATC GCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAG AGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGGTGGGACAGATGATTGGGGTGA AGTCGTAACAAGGTAGCCGTATCGGAAGGTGCGGCTGGATCACCTCCTTT (SEQ ID NO : 2 )

[0144] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 3

[0145] (also referred to herein as “Strain 3”) has the highest homology with a bacterial strain of the species Bacillus pumilus:

[0146] Strain 3 16S ribosomal RNA coding sequence (16S rDNA)

[0147] GAGT T TGATCCTGGCT C AGGAC GAAC GCTGGCGGCGTGCCTAATACAT GC AAGT C GAGC GAAC AGAAGGGAGC T T GCTCCCGGATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAA AC C GGAGC T AAT AC C GGAT AGT T C C T T GAAC C GC AT GGT T C AAGGAT GAAAGAC GGT T T C GGC T GT C AC T T AC AG ATGGACCCGCGGCGCATTAGCTAGTTGGTGGGGTAATGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGG GTGATCGGCCACACT GGGAC T GAGAC AC GGC C C AGAC T C C T AC GGGAGGC AGC AGT AGGGAAT CTTCCGCAATGG ACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAA C AAGT GC GAGAGT AAC T GC T C GC AC C T T GAC GGT AC C T AAC C AGAAAGC C AC GGC T AAC T AC GT GC C AGC AGC C G CGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTTCTTAAGTCTGA T GT GAAAGC CCCCGGCT C AAC C GGGGAGGGT CAT T GGAAAC T GGGAAAC T T GAGT GC AGAAGAGGAGAGT GGAAT TCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTG ACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCT AAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAA GACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAG AAC CTTACCAGGTCTT GAC AT C C T C T GAC AAC C C T AGAGAT AGGGC TTTCCCTTC GGGGAC AGAGT GAC AGGT GG TGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTT GCCAGCATTTAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCAT C AT GC C C C T TAT GAC C T GGGC T AC AC AC GT GC T AC AAT GGAC AGAAC AAAGGGC T GC GAGAC C GCAAGGT T T AGC PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0148] CAATCCCATAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATC GCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGCAAC ACCCGAAGTCGGTGAGGTAACCTTTATGGAGCCAGCCGCCGAAGGTGGGGCAGATGATTGGGGTGAAGTCGTAAC AAGGTAGCCGTATCGGAAGGTGCGGCTGGATCACCTCCTTT (SEQ ID NO : 3 )

[0149] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 4

[0150] (also referred to herein as “Strain 4”) has the highest homology with a bacterial strain of the species Bacillus subtilis:

[0151] Strain 4 16S ribosomal RNA coding sequence (16S rDNA)

[0152] T C GGAGAGT T TGATCCTGGCT C AGGAC GAAC GCTGGCGGCGTGCCTAATACAT GC AAGT C GAGC GGAC AGAT GGG AGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCC GGGAAACCGGGGCTAATACCGGATGGTTGTTTGAACCGCATGGTTCAAACATAAAAGGTGGCTTCGGCTACCACT TACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTG AGAGGGT GAT C GGC C AC AC T GGGAC T GAGAC AC GGC C C AGAC T C C T AC GGGAGGC AGC AGT AGGGAAT C T T C C GC AATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGG AAGAACAAGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTTCTTAA GT C T GAT GT GAAAGC CCCCGGCT C AAC C GGGGAGGGT C AT T GGAAAC T GGGGAAC T T GAGT GC AGAAGAGGAGAG TGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTG T AAC T GAC GC T GAGGAGC GAAAGC GT GGGGAGC GAAC AGGAT T AGAT AC CCTGGTAGTCCACGC C GT AAAC GAT G AGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGG TCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAAC GC GAAGAAC CTTACCAGGTCTT GAC AT C C T C T GAC AAT C C TAGAGAT AGGAC GT C C C C T T C GGGGGC AGAGT GAC AGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATC TTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCA AAT C AT C AT GC C C C T T AT GAC C T GGGC T AC AC AC GT GC T AC AAT GGAC AGAAC AAAGGGC AGC GAAAC C GC GAGG TTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTA GTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTT TGTAACACCCGAAGTCGGTGAGGTAACCTTTTAGGAGCCAGCCGCCGAAGGTGGGACAGATGATTGGGGTGAAGT CGTAACAAGGTAGCCGTATCGGAAGGTGCGGCTGGATCACCTCCTTT (SEQ ID NO : 4 )

[0153] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 5

[0154] (also referred to herein as “Strain 5”) has the highest homology with a bacterial strain of the species Bacillus velezensis:

[0155] Strain 5 16S ribosomal RNA coding sequence (16S rDNA)

[0156] ATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGC TAATACCGGATGGTTGTCTGAACCGCATGGTTCAGACATAAAAGGTGGCTTCGGCTACCACTTACAGATGGACCC GCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGG C C AC AC T GGGAC T GAGAC AC GGC C C AGAC T C C T AC GGGAGGC AGC AGT AGGGAAT CTTCCGCAAT GGAC GAAAGT CTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCC GTTCAAATAGGGCGGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAAT ACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAA GCCCCCGGCT C AAC C GGGGAGGGT CAT T GGAAAC T GGGGAAC T T GAGT GC AGAAGAGGAGAGT GGAAT T C C AC GT GTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGA GGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTT AGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAA ACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTA CCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGG TTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCA TTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCC C T TAT GAC C T GGGC T AC AC AC GT GC T AC AAT GGAC AGAAC AAAGGGC AGC GAAAC C GC GAGGT TAAGC C AAT C C C ACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATC AGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAA PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0157] GTCGGTGAGGTAACCTTTATGGAGCCAGCCGCCGAAGGTGGGACAGATGATTGGGGTGAAGTCGTAACAAGGTAG CCGTATCGGAAGGTGCGGCTGGATCACCTCCTTT (SEQ ID NO : 5 )

[0158] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 6

[0159] (also referred to herein as “Strain 6”) has the highest homology with a bacterial strain of the species Weizmannia coagulans:

[0160] Strain 6 16S ribosomal RNA coding sequence (16S rDNA)

[0161] TCACTTACAGATGGGCCCGCGGCGCATTAGCTAGTTGGTGGGGTAACGGCTCACCAAGGCAACGATGCGTAGCCG ACCT GAGAGGGT GAT CGGCCACATT GGGAC T GAGAC AC GGC C C AAAC T C C T AC GGGAGGC AGC AGT AGGGAAT C T TCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGAAGAAGGCCTTCGGGTCGTAAAACTCTGTTGC C GGGGAAGAAC AAGT GC C GT T C GAAC AGGGC GGC GC C T T GAC GGT AC C C GGC C AGAAAGC C AC GGC T AAC T AC GT GCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTT CTTAAGTCTGATGTGAAATCTTGCGGCTCAACCGCAAGCGGTCATTGGAAACTGGGAGGCTTGAGTGCAGAAGAG GAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGGCTCTCTG GTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAA CGATGAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAG TACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAA GC AAC GC GAAGAAC CTTACCAGGTCTT GAC AT C C T C T GAC C T C C C T GGAGAC AGGGC CTTCCCCTTC GGGGGAC A GAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACC CTTGACCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGAT GACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAAAGGGCTGCGAGAC C GC GAGGT T AAGC C AAT C C C AGAAAAC C AT T C C C AGT T C GGAT T GC AGGC T GC AAC C C GC C T GC AT GAAGC C GGA ATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCAC GAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTT (SEQ ID NO : 6)

[0162] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 7

[0163] (also referred to herein as “Strain 7”) has the highest homology with a bacterial strain of the species Lactiplantibacillus plantarum'.

[0164] Strain 7 16S ribosomal RNA coding sequence (16S rDNA)

[0165] T T T GAGAGT T T GAT C C T GGC T C AGGAC GAAC GC T GGC GGC GT GC C T AAT AC AT GC AAGT C GAAC GAAC T C T GGT A TTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCA GAAGC GGGGGAT AAC AC C T GGAAAC AGAT GC T AAT AC C GC AT AAC AAC T T GGAC CGCATGGTCC GAGT T T GAAAG ATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGAGGTAACGGCTCACCATGGCAA T GAT AC GT AGC C GAC C T GAGAGGGT AAT CGGCCACATT GGGAC T GAGAC AC GGC C C AAAC T C C T AC GGGAGGC AG CAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGT AAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCAC GGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAG CGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTG AGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAA GGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGT CCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATT C C GC C T GGGGAGT AC GGC C GC AAGGC T GAAAC T C AAAGGAAT T GAC GGGGGC C C GC AC AAGC GGT GGAGC AT GT G GTTTAATTC GAAGC T AC GC GAAGAAC CTTACCAGGTCTT GAC AT AC TAT GC AAAT C T AAGAGAT T AGAC GT T C C C TTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAAC GAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGA AGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGA GTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACA TGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCC GTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGGTGGGACA GATGATTAGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCGGCTGGATCACCTCCTT (SEQ ID NO : 7 ) PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0166] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 15

[0167] (also referred to herein as “Strain 8”) has the highest homology with a bacterial strain of the species Bifidobacterium adolescentis'.

[0168] Strain 8 16S ribosomal RNA coding sequence (16S rDNA)

[0169] NNNNTTGTGGAGGGTTCGATTCTGGCTCAGGATNAACGCTNGCGGCGTGCTTAACACATGCAAGTCGAACGGGAT CGGCTNGAGCTTGCTCCGGCTGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCGACCTGCCCCATACACCGGA ATAGCTCCTGGAAACGGGTGGTAATGCCGGATGCTCCAGTTGGATGCATGTCCTTCTGGGAAAGATTCTATCGGT ATGGGATGGGGTCGCGTCCTATCAGCTTGATGGCGGGGTAACGGCCCNCCATGGCTTCGACGGGNAGCCGGCCTG AGAGGGC GAC CGGCCACATT GGGAC T GAGAT AC GGC C CNGAC T C C T AC GGGAGGC AGC AGT GGGNAAT AT T GC AC AATGGGCGCAAGCCTAATGCAGCGACGCCGCGTGCGGGATGACGGCCTTCGGGTTGTAAACCGCTTTTGACTGGG AGCAAGCCTTCGGGGTGAGTGTACCTTTCGAATAAGCACCGGCTAACTACGTGCCAGCAGCCNCGGTAATACGTA GGGTGCNAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCA TCGCTTAACGGTGGNTCCGCGCCGGGTACGGGCGGNCTTGAGTGCGGTAGGGNAGACTGGAATTCCNGGTGTAAC GGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCNGTNACTGACGCTGAGGAGC GAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATGCTGGATGTGGGGAC CATTCCACGGTCTCCGTGTCGGAGCCAACGCGTTAAGCATCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTC AAAGAAAT T GAC GGGNNC CNNC AC AAGC GGCNGAGC AT GCGGATTAATTC GATNNAAC GC GAAGAAC C T T AC C T G GGCTTGACATGTTCCCGACAGGCCCCAGAGATGGGNNNTCCTTCGGGNCGGGNTCACAGGTGGNGCATGGTCGTC GTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTGTGTTGCCAGCACGTCG TGGTGGNAACTCACGGGNGACCGCCGGGGTCAACTCGGAGGAAGGTGGGNATGACGTCAGATCATCATGCCCCTT ACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAACGGGATGCGACCTCGTGAGGGGGAGCGGATCCCTTA AAACCGGNCTCAGTTCGGATTGGAGTCTGCAACCCGACTCCATGAAGGCGGAGTCGCTAGTAATCGCGGATCAGC AACGCCGCGGTNAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGCCATGAAAGTGGGTAGCACCCGAAGC CGGTGGCCCNACCTTTTTGGGGGGAGCCGTCTAAGGTGAGNCTCGTGATNGG (SEQ ID NO : 15 )

[0170] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 16 (also referred to herein as “Strain 9”) has the highest homology with a bacterial strain of the species Bifidobacterium bifidum'.

[0171] Strain 9 16S ribosomal RNA coding sequence (16S rDNA)

[0172] TTTTTGTGGAGGGTTCGATTCTGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGGATC CATCAAGCTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCGACCTGCCCCATGCTCCGGAATA GCTCCTGGAAACGGGTGGTAATGCCGNATGTTCCACATGATCGCATGTGATTGTGGGAAAGATTCTATCGGCGTG GGATGGGGTCGNGTCCTATCAGCTTGTTGGTGAGGTAACGGCTCACCAAGGCTTCGACGGGTAGCCGGCCTGAGA GGGC GAC C GGC C AC AT T GGGAC T GAGAT AC GGC C CAGAC T C C T AC GGGAGGC AGC AGT GGGGAAT AT T GC AC AAT GGGCGCAAGCCTGATGCAGCGACGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTGTTTGGGAGC AAGCCTTCGGGTGAGTGTACCTTTCGAATAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGN NNNAGCGTTATCCGGATTTATTGGGCGTAAAGGGCTCGTAGGCGGCTCGTCGCGTCCGGTGTGAAAGTCCATCGC TTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTG GAATGTGTAGATATCGGGAAGAACACCGATGGCGAAGGCAGGTCTCTGGGCNGTCACTGACGCTGAGGAGCNAAA GCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGACGCTGGATGTGGGGCACGTT CCACGTGTTCCGTGTCGGAGCTAACGCGTTAAGCGTCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAG AAATTGACGGGGGCCNGCACAAGCGGCGGAGCATGCGGATTAATTCGAACNAACGCGAAGAACCTTACCTGGGCT TGACATGTTCCCGACGACGCCAGAGATGGCGTTTCCCTTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCA GCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCACGTTATGGT GGGAACTCACGGGNNACCGCCGGGGTTAACNCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGT C C AGGGC T T C AC GC AT GC T AC AAT GGC C GGT AC AGC GGGAT GC GAC AT GGC GAC AT GGAGC GGAT C C C T GAAAAC CGGTCTCAGTTCGGATCGGAGCCTGCAACCCGGCTCCGTGAAGGCGGAGTCGCTAGTAATCGCGGATCAGCAACG CCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAGCCGGT GGCCTAACCCCTTGTGGGATGGAGCCGTCTAAGGTGAGGCTCGTGNTTGGGACTAAGNNGTAACAAGNNNNNNGT ACCGGAAGNNNNNNNNNGATCACCTCCTTTCT (SEQ ID NO : 16) PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0173] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 17 (also referred to herein as “Strain 10”) has the highest homology with a bacterial strain of the species Bifidobacterium longum'.

[0174] Strain 10 16S ribosomal RNA coding sequence (16S rDNA)

[0175] GATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGGGATCCATCAAGCTTGCTTGGTGGTGAGAGTGGC GAACGGGTGAGTAATGCGTGACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGAT GTTCCAGTTGATCGCATGGTCTTCTGGGAAAGCTTTCGCGGTATGGGATGGGGTCGCGTCCTATCAGCTTGACGG CGGGGTAACGGCCCACCGTGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATAC GGCCCATACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGT GAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGGAGCAAGCGAGAGTGAGTTTACCCGTTGAATAAG CACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTATCCGGAATTATTGGGCGTAAAG GGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGG CTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGG C GAAGGC AGGT C TCTGGGCCGTTACT GAC GC T GAGGAGC GAAAGC GT GGGGAGC GAAC AGGAT T AGAT AC C C T GG TAGTCCACGCCGTAAACGGTGGATGCAGGATGTGGGGCCCGTTCCACGGGTTCCGTGTCGGAGCTAACGCGTTAA GCATCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGC ATGCGGATTAATTCGAT GC AAC GC GAAGAAC CTTACCTGGGCTT GAC AT GT T C C C GAC GGT C GT AGAGAT AC GGC TTCCCTTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCG CAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCGGATTATGCCGGGAACTCACGGGGGACCGCCGGGGTTAACTC GGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTA C AAC GGGAT GC GAC GC GGC GAC GC GGAGC GGAT C C C T GAAAAC C GGT C T C AGT T C GGAT C GC AGT C T GC AAC T C G ACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGAATCAGCAACGTCGCGGTGAATGCGTTCCCGGGCCTTGTACAC ACTGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAGCCGGTGGCCTAACCCCTTGTGGGATGGAGCCGTCTAA GGTGAGGCTCGTGATTGGGACTAAGTCGTAACAAGGTAGCCGTACCGGAAGG (SEQ ID NO : 17 )

[0176] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 18 (also referred to herein as “Strain 11”) has the highest homology with a bacterial strain of the species Lactobacillus acidophilus'.

[0177] Strain 11 16S ribosomal RNA coding sequence (16S rDNA)

[0178] NNAAAAC GAGAGT T TGATCCTGGCT C AGGAC GAAC GCTGGCGGCGTGCCTAATACAT GC AAGT C GAGC GAGC T GA ACCAACAGATTCACTTCGGTGATGACGTTGGGNAACGCTAGCGGCGGATGGGTGAGTAACACGTGGGGAACCTGC C C CAT AGT C T GGGAT AC C AC T T GGAAAC AGGT GC TAAT AC C GGAT AAGAAAGC AGAT C GC AT GAT CAGC T TAT AA AAGGCGGCGTAAGCTGTCGCTATGGNNTGGCCCCGCGGTGCATTAGCTAGTTGGTAGGGTAACGGCCTACCAAGG CAATGATGCATAGCC GAGT T GAGAGAC TGATCGGCCACATT GGGAC T GAGAC AC GGC C C AAAC T C C T AC GGGAGG CAGCNGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGAT CGTAAAGCTCTGTTGTTGGTGAAGAAGGATAGAGGTAGTAACTGGCCTTTATTTGACGGTAATCAACCAGAAAGT CACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCNAGCGTTGTCCGGATTTATTGGGCGTAAAGC GAGC GC AGGC GGAAGAAT AAGT C T GAT GT GAAAGC CCTCGGCT T AAC C GAGGAAC T GC AT C GGAAAC TGTTTTTC TTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGC GAAGGCGGCTCTCTGGTCTGCAACTGACGCTGAGGCTCNNAAGCATGGGTAGCGAACAGGATTAGATACCCTGGT AGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGC AC T C C GC C T GGGGAGT AC GAC C GC AAGGT T GAAAC T C AAAGGAAT T GAC GGGGNC C C GC AC AAGC GGT GGAGC AT GTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTAGTGCAATCCGTAGAGATACGGNGTT CCCTTCGGGGACACTAAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGC AACGAGTGCAACCCTTGTCATTAGTTGCCAGCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGA GGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGTACAA C GAGGAGC AAGC C T GC GAAGGC AAGC GAAT C T C T T AAAGC TGTTCTCAGTTC GGAC T GC AGT C T GC AAC T C GAC T GCACGAAGCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCG CCCGTCACACCATGGGAGTCTGCAATGCCCAAAGCCGGTGGCCTAACCTTCGGGAAGGAGCCGTCTAAGGCAGGG C AGAT GACNNNNNNNNNNNNGT AAC AAGNNNNNNNNNNNNGAAC C T GNNNNNNGAT CACCTCCTTTCTA (SEQ ID NO : 18 ) PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0179] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 19 (also referred to herein as “Strain 12”) has the highest homology with a bacterial strain of the species Lactobacillus delbrueckiv.

[0180] Strain 12 16S ribosomal RNA coding sequence (16S rDNA)

[0181] NT C AAAT T GAGAGT T T GAT C C T GGC T C AGGAC GAAC GC T GGC GGC GT GC C T AAT AC AT GC AAGT C GAGC GAGC T G AATTCAAAGATCCCTTCGGGGTGATTTGTTGGATGCTAGCGGCGGATGGGTGAGTAACACGTGGGCAATCTGCCC T AAAGAC T GGGAT AC C AC T T GGAAAC AGGT GC T AAT AC C GGAT AAC AAC AT GAAT CGCATGATT C AAGT T T GAAA GGCGGCGYAAGCTGTCACTTTAGGATGAGCCCGCGGCGCATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCA ATGATGCGTAGCC GAGT T GAGAGAC TGATCGGCCACATT GGGAC T GAGAC AC GGC CNNAAC T C C T AC GGGAGGC A GCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTCTTCGGATCG TAAAGCTCTGTTGTTGGTGAAGAAGGATAGAGGCAGTAACTGGTCTTTATTTGACGGTAATCAACCAGAAAGTCA CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGA GCGCAGGCGGAATGATAAGTCTGATGTGAAAGCCNACGGCTYAACCGTGGAACTGCATCGGAAACTGTCATTCTT GAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGA AGGC GGC T C T C T GGT C T GC AAC T GAC GC T GAGGC T C GAAAGC AT GGGT AGC GAAC AGGAT T AGAT AC C C T GGT AG TCCATGCCGTAAACGATGAGCGCTAGGTGTTGGGGACTTTCCAGTCCTCAGTGCCGCAGCAAACGCATTAAGCNN TCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCNGCACAAGCGGTGGAGCATGT GGTTTAATTCGAAGNAACGCGAAGAACCTTACCAGGTCTTGACATCCTGCGCTACACCTAGAGATAGGTNGTTCC CTTCGGGGACGCAGAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAA C GAGC GC AAC C C T T GT C T T T AGT T GC C AT C AT T AAGT T GGGC AC T C T AAAGAGAC T GC C GGT GAC AAAC C GGAGG AAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGTACAACG AGAAGC GAAC C C GC GAGGGTAAGC GGAT C T C T TAAAGC T GY T C T C AGT T C GGAC T GC AGGC T GC AAC T C GC C T GC ACGAAGCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCC CGTCACACCATGGAAGTCTGCAATGCCCAAAGTCGGTGAGATAACCTTTATAGGAGTCAGCCGCCTAAGGCAGGG CAGATGACNGGG (SEQ ID NO : 19)

[0182] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 20 (also referred to herein as “Strain 13”) has the highest homology with a bacterial strain of the species Lactobacillus helveticus'.

[0183] Strain 13 16S ribosomal RNA coding sequence (16S rDNA)

[0184] AC GAAC GC T GGC GGC GT GC C T AAT AC AT GC AAGT C GAGC GAGC AGAAC C AGC AGAT T T AC T T C GGT AAT GAC GC T GGGGACGCGAGCGGCGGATGGGTGAGTAACACGTGGGGAACCTGCCCCATAGTCTGGGATACCACTTGGAAACAG GTGCTAATACCGGATAAGAAAGCAGATCGCATGATCAGCTTATAAAAGGCGGCGTAAGCTGTCGCTATGGGATGG CCCCGCGGTGCATTAGCTAGTTGGTAAGGTAACGGCTTACCAAGGCAATGATGCATAGCCGAGTTGAGAGACTGA TCGGCCACATT GGGAC T GAGAC AC GGC C C AAAC T C C T AC GGGAGGC AGC AGT AGGGAAT CTTCCACAAT GGAC GC AAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTGGTGAAGAAGGAT AGAGGTAGTAACTGGCCTTTATTTGACGGTAATCAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGT AATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGAAGAATAAGTCTGATGTG AAAGC CCTCGGCT T AAC C GAGGAAC T GC AT C GGAAAC TGTTTTTCTT GAGT GC AGAAGAGGAGAGT GGAAT T C C A TGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGCAACTGACGC TGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGT GTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTT GAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACC TTACCAGGTCTTGACATCTAGTGCCATCCTAAGAGATTAGGAGTTCCCTTCGGGGACGCTAAGACAGGTGGTGCA TGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTATTAGTTGCCA GCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATG CCCCTTAT GAC CTGGGCTACACACGTGCTACAAT GGAC AGT AC AAC GAGAAGC GAGC C T GC GAAGGC AAGC GAAT CTCTGAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACTCGACTGCACGAAGCTGGAATCGCTAGTAATCGCGG ATCAGAACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGAAGTCTGCAATGCCC AAAGCCGGTGGCCTAACCTTCGGGAAGGAGCCGTCTAAGGCAGGGCAGATGACTGGGGTGAA (SEQ ID NO : 20 ) PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0185] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 21

[0186] (also referred to herein as “Strain 14”) has the highest homology with a bacterial strain of the species Lactobacillus reuteri'.

[0187] Strain 14 16S ribosomal RNA coding sequence (16S rDNA)

[0188] AGAGTTTGATNNTGGCTCAGGATGAACGCCGGCGGTGTGCCTAATACATGCAAGTCGTACGCACTGGCCCAACTG ATTGATGGTGCTTGCACCTGATTGACGATGGATCACCAGTGAGTGGCGGACGGGTGAGTAACACGTAGGTAACCT GCCCCGGAGCGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAACAACAAAAGCCGCATGGCTTTTGTTT GAAAGATGGCTTTGGCTATCACTCTGGGATGGACCTGCGGTGCATTAGCTAGTTGGTAAGGTAACGGCTTACCAA GGCGATGATGCATAGCC GAGT T GAGAGAC TGATCGGCCACAAT GGAAC T GAGAC AC GGTCCATACTCCTACGGGA GGCAGCAGTAGGGAATCTTCCACAATGGGCGCAAGCCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGG CTCGTAAAGCTCTGTTGTTGGAGAAGAACGTGCGTGAGAGTAACTGTTCNCGCAGTGACGGTATCCAACCAGAAA GTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAA GCGAGCGCAGGCGGTTGCTTAGGTCTGATGTGAAAGCCTTCGGCTTAACCGAAGAAGTGCATCGGAAACCGGGCG ACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTG GC GAAGGC GGC T GT C T GGT C T GC AAC T GAC GC T GAGGC T C GAAAGC AT GGGT AGC GAAC AGGAT T AGAT AC C C T G GTAGTCCATGCCGTAAACGATGAGTGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGGAGCTAACGCATTAA GC AC T C C GC C T GGGGAGT AC GAC C GC AAGGT T GAAAC T C AAAGGAAT T GAC GGGGGC C C GC AC AAGC GGT GGAGC ATGTGGTTTAATTC GAAGC T AC GC GAAGAAC CTTACCAGGTCTT GAC AT C T T GC GC T AAC C T T AGAGAT AAGGC G TTNCCTTCGGGGACGCAATGACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCT GCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCG GAGGAAGGTGGGGACGACGTCAGATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTAC AACGAGTCGCAAGCTCGCGAGAGTAAGCTAATCTCTTAAAGCCGTTCTCAGTTCGGACTGTAGGCTGCAACTCGC CTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACAC CGCCCGTCACACCATGGGAGTTTGTAACGCCCAAAGTCGGTGGCCTAACCTTTATGGAGGGAGCCGCCTAAGGCG GGACAGATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAGCCTGCGGCTGGATCACCTCCTTTNT (SEQ ID NO : 21 )

[0189] The bacterial strain comprising a 16S rDNA sequence provided by SEQ ID NO: 22 (also referred to herein as “Strain 15”) has the highest homology with a bacterial strain of the species Lactococcus lactis:

[0190] Strain 15 16S ribosomal RNA coding sequence (16S rDNA)

[0191] NNTTATTTGAGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTTGAGCGCTGAA GGTTGGTACTTGTACCGACTGGATGAGCAGCGAACGGGTGAGTAACGCGTGGGGAATCTGCCTTTGAGCGGGGGA CAACATTTGGAAACGAATGCTAATACCGCATAAAAACTTTAAACACAAGTTTTAAGTTTGAAAGATGCAATTGCA TCACTCAAAGATGATCCCGCGTTGTATTAGCTAGTTGGTGAGGTAAAGGCTCACCAAGGCGATGATACATAGCCG ACCT GAGAGGGT GAT CGGCCACATT GGGAC T GAGAC AC GGC C C AAAC T C C T AC GGGAGGC AGC AGT AGGGAAT C T TCGGCAATGGACGAAAGTCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAACTCTGTTGG TAGAGAAGAACGTTGGTGAGAGTGGAAAGCTCATCAAGTGACGGTAACTACCCAGAAAGGGACGGCTAACTACGT GCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGTGGTTT ATTAAGTCTGGTGTAAAAGGCAGTGGCTCAACCATTGTATGCATTGGAAACTGGTAGACTTGAGTGCAGGAGAGG AGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGG CCTGTAACTGACACTGAGGCTCNNAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAAC GATGAGTGCTAGATGTAGGGAGCTATAAGTTCTCTGTATCGCAGCTAACGCAATAAGCACTCCGCCTGGGGAGTA CGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCNGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGC AAC GC GAAGAAC CTTACCAGGTCTT GAC AT AC TCGTGCTATTCC T AGAGAT AGGAAGT T C C T T C GGGAC AC GGGA TACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTA TTGTTAGTTGCCATCATTAAGTTGGGCACTCTAACGAGACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACG TCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTCGCGAGACAGTG ATGTTTAGCTAATCTCTTAAAACCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCG CTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGGGA GTTGGGAGTACCCGAAGTAGGTTGCCTAACCGCAAGGAGGGCGCTTCCTAAGGTAAGACCGATGACNNGGGNNNN NNGTAACAAGNNNNNNNGTATCGGAAGNNNNNNNNNGATCACCTCCTTTCTA (SEQ ID NO : 22 ) PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0192] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Pediococcus acidilactici, wherein the bacterial strains belonging to the species Pediococcus acidilactici are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Pediococcus acidilactici. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Pediococcus acidilactici. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Pediococcus acidilactici. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Pediococcus acidilactici has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Pediococcus acidilactici.

[0193] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bacillus licheniformis, wherein the bacterial strains belonging to the species Bacillus licheniformis are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bacillus licheniformis. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bacillus licheniformis. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Bacillus licheniformis. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bacillus licheniformis has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bacillus licheniformis.

[0194] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bacillus pumilus, wherein the bacterial strains belonging to the species Bacillus pumilus are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bacillus pumilus. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bacillus pumilus. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Bacillus pumilus. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bacillus pumilus has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0195] 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bacillus pumilus.

[0196] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bacillus subtilis. wherein the bacterial strains belonging to the species Bacillus subtilis are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bacillus subtilis. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bacillus subtilis. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Bacillus subtilis. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bacillus subtilis has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bacillus subtilis.

[0197] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bacillus velezensis, wherein the bacterial strains belonging to the species Bacillus velezensis are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bacillus velezensis. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bacillus velezensis. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Bacillus velezensis. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bacillus velezensis has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bacillus velezensis.

[0198] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Weizmannia coagulans, wherein the bacterial strains belonging to the species Weizmannia coagulans are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Weizmannia coagulans. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Weizmannia coagulans. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Weizmannia coagulans. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Weizmannia coagulans has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0199] 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Weizmannia coagulans.

[0200] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Lactiplantibacillus plantarum, wherein the bacterial strains belonging to the species Lactiplantibacillus plantarum are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Lactiplantibacillus plantarum. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Lactiplantibacillus plantarum. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Lactiplantibacillus plantarum. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Lactiplantibacillus plantarum has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Lactiplantibacillus plantarum.

[0201] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bifidobacterium adolescentis, wherein the bacterial strains belonging to the species Bifidobacterium adolescentis are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bifidobacterium adolescentis. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bifidobacterium adolescentis. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Bifidobacterium adolescentis. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bifidobacterium adolescentis has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bifidobacterium adolescentis.

[0202] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bifidobacterium bifidum, wherein the bacterial strains belonging to the species Bifidobacterium bifidum are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bifidobacterium bifidum. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bifidobacterium bifidum. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0203] Bifidobacterium bifidum. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bifidobacterium bifidum has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bifidobacterium bifidum.

[0204] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Bifidobacterium longum, wherein the bacterial strains belonging to the species Bifidobacterium longum are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Bifidobacterium longum. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Bifidobacterium longum. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Bifidobacterium longum. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Bifidobacterium longum has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Bifidobacterium longum.

[0205] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Lactobacillus acidophilus, wherein the bacterial strains belonging to the species Lactobacillus acidophilus are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Lactobacillus acidophilus. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Lactobacillus acidophilus. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Lactobacillus acidophilus. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Lactobacillus acidophilus has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Lactobacillus acidophilus.

[0206] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Lactobacillus delbrueckii, wherein the bacterial strains belonging to the species Lactobacillus delbrueckii are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Lactobacillus delbrueckii. In some embodiments, the composition comprises fermentates of PCT / US25 / 16818 21 February 2025 (21.02.2025) two bacterial strains belonging to Lactobacillus delbrueckii. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Lactobacillus delbrueckii. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Lactobacillus delbrueckii has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Lactobacillus delbrueckii.

[0207] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Lactobacillus helveticus, wherein the bacterial strains belonging to the species Lactobacillus helveticus are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Lactobacillus helveticus. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Lactobacillus helveticus. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Lactobacillus helveticus. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Lactobacillus helveticus has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Lactobacillus helveticus.

[0208] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Lactobacillus reuteri, wherein the bacterial strains belonging to the species Lactobacillus reuteri are different bacterial strains. In some embodiments, the composition comprises a fermentate of one bacterial strain belonging to Lactobacillus reuteri. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Lactobacillus reuteri. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Lactobacillus reuteri. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Lactobacillus reuteri has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Lactobacillus reuteri.

[0209] In some embodiments, a composition comprises fermentates of two or more bacterial strains belonging to the species Lactococcus laclis, wherein the bacterial strains belonging to the species Lactococcus lactis are different bacterial strains. In some embodiments, the PCT / US25 / 16818 21 February 2025 (21.02.2025) composition comprises a fermentate of one bacterial strain belonging to Lactococcus lactis. In some embodiments, the composition comprises fermentates of two bacterial strains belonging to Lactococcus lactis. In some embodiments, the composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains belonging to Lactococcus lactis. In some embodiments, the 16S rDNA sequence of the first bacterial strain belonging to Lactococcus lactis has no more than 99.9%, 99.8%, 99.7%, 99.6%, 99.5%, 99.4%, 99.3%, 99.2%, 99.1%, 99.0%, 98.8%, 98.6%, 98.5%, 98.4%, 98.2%, 98.0%, 97.8%, 97.6%, 97.5%, 97.4%, or 97.2% identity to 16S rDNA sequence of the second bacterial strain belonging to Lactococcus lactis.

[0210] In some embodiments, a composition comprises a fermentate of one or more additional bacterial strains, in addition to the first fermentate comprising ILA and the second fermentate comprising lAld. Additional bacterial strains that produce fermentates may be taxonomically or phylogenetically related to bacterial strains of Pediococcus acidilactici, Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans, and Lactiplantibacillus plantarum. In some embodiments, one or more additional bacterial strains belong to the species Pediococcus acidilactici. In some embodiments, one or more additional bacterial strains belong to the species Bacillus licheniformis. In some embodiments, one or more additional bacterial strains belong to the species Bacillus pumilus. In some embodiments, one or more additional bacterial strains belong to the species Bacillus subtilis. In some embodiments, one or more additional bacterial strains belong to the species Bacillus velezensis. In some embodiments, one or more additional bacterial strains belong to the species Weizmannia coagulans. In some embodiments, one or more additional bacterial strains belong to the species Lactiplantibacillus plantarum. In some embodiments, one or more additional bacterial strains belong to the species Bifidobacterium adolescentis. In some embodiments, one or more additional bacterial strains belong to the species Bifidobacterium bifidum. In some embodiments, one or more additional bacterial strains belong to the species Bifidobacterium longum. In some embodiments, one or more additional bacterial strains belong to the species Lactobacillus acidophilus. In some embodiments, one or more additional bacterial strains belong to the species Lactobacillus delbrueckii. In some embodiments, one or more additional bacterial strains belong to the species Lactobacillus helveticus. In some embodiments, one or more additional bacterial strains belong to the species Lactobacillus reuteri. In some embodiments, one or more additional bacterial strains belong to the species Lactococcus lactis. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0211] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Pediococcus acidilactici. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Pediococcus acidilactici is not identical to SEQ ID NO: 1. In some embodiments, an additional bacterial strain belonging to Pediococcus acidilactici comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 1. In some embodiments, each additional bacterial strain belonging to the species Pediococcus acidilactici comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%,

[0212] 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%,

[0213] 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%,

[0214] 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 8. In some embodiments, each additional bacterial strain belonging to the species Pediococcus acidilactici comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 8.

[0215] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bacillus licheniformis . In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Bacillus licheniformis is not identical to SEQ ID NO: 2. In some embodiments, an additional bacterial strain belonging to Bacillus licheniformis comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 2. In some embodiments, each additional bacterial strain belonging to the species Bacillus licheniformis comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%,

[0216] 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%,

[0217] 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%,

[0218] 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 9. In some embodiments, each additional bacterial strain belonging to the species Bacillus licheniformis comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 9. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0219] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bacillus pumilus. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Bacillus pumilus is not identical to SEQ ID NO: 3. In some embodiments, an additional bacterial strain belonging to Bacillus pumilus comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 3. In some embodiments, each additional bacterial strain belonging to the species Bacillus pumilus comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%,

[0220] 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%,

[0221] 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%,

[0222] 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%

[0223] ANI to the reference genome of SEQ ID NO: 10. In some embodiments, each additional bacterial strain belonging to the species Bacillus pumilus comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 10.

[0224] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bacillus subtilis. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Bacillus subtilis is not identical to SEQ ID NO: 4. In some embodiments, an additional bacterial strain belonging to Bacillus subtilis comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 4. In some embodiments, each additional bacterial strain belonging to the species Bacillus subtilis comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%,

[0225] 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%,

[0226] 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%,

[0227] 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%

[0228] ANI to the reference genome of SEQ ID NO: 11. In some embodiments, each additional bacterial strain belonging to the species Bacillus subtilis comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 11. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0229] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bacillus velezensis. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Bacillus velezensis is not identical to SEQ ID NO: 5. In some embodiments, an additional bacterial strain belonging to Bacillus velezensis comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 5. In some embodiments, each additional bacterial strain belonging to the species Bacillus velezensis comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%,

[0230] 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%,

[0231] 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%,

[0232] 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%

[0233] ANI to the reference genome of SEQ ID NO: 12. In some embodiments, each additional bacterial strain belonging to the species Bacillus velezensis comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 12.

[0234] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Weizmannia coagulans. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Weizmannia coagulans is not identical to SEQ ID NO: 6. In some embodiments, an additional bacterial strain belonging to Weizmannia coagulans comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 6. In some embodiments, each additional bacterial strain belonging to the species Weizmannia coagulans comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97.0%, 97.1%, 97.2%,

[0235] 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%,

[0236] 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%,

[0237] 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 13. In some embodiments, each additional bacterial strain belonging to the species Weizmannia coagulans comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 13. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0238] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Lactiplantibacillus plantarum. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Lactiplantibacillus plantarum is not identical to SEQ ID NO: 7. In some embodiments, an additional bacterial strain belonging to Lactiplantibacillus plantarum comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 7. In some embodiments, each additional bacterial strain belonging to the species Lactiplantibacillus plantarum comprises a genome with at least 95.0%, 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96.0%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%,

[0239] 96.8%, 96.9%, 97.0%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%,

[0240] 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%,

[0241] 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% ANI to the reference genome of SEQ ID NO: 14. In some embodiments, each additional bacterial strain belonging to the species Lactiplantibacillus plantarum comprises a genome with an AF of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to the reference genome of SEQ ID NO: 14.

[0242] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bifidobacterium adolescentis. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Bifidobacterium adolescentis is not identical to SEQ ID NO: 15. In some embodiments, an additional bacterial strain belonging to Bifidobacterium adolescentis comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 15.

[0243] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bifidobacterium bifidum. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Bifidobacterium bifidum is not identical to SEQ ID NO: 16. In some embodiments, an additional bacterial strain belonging to Bifidobacterium bifidum comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 16.

[0244] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Bifidobacterium longum. In some embodiments, the 16S rDNA sequence of an PCT / US25 / 16818 21 February 2025 (21.02.2025) additional bacterial strain belonging to Bifidobacterium longum is not identical to SEQ ID NO: 17. In some embodiments, an additional bacterial strain belonging to Bifidobacterium longum comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 17.

[0245] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Lactobacillus acidophilus. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Lactobacillus acidophilus is not identical to SEQ ID NO: 18. In some embodiments, an additional bacterial strain belonging to Lactobacillus acidophilus comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 18.

[0246] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Lactobacillus delbrueckii. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Lactobacillus delbrueckii is not identical to SEQ ID NO: 19. In some embodiments, an additional bacterial strain belonging to Lactobacillus delbrueckii comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 19.

[0247] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Lactobacillus helveticus. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Lactobacillus helveticus is not identical to SEQ ID NO: 20. In some embodiments, an additional bacterial strain belonging to Lactobacillus helveticus comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 20.

[0248] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the species Lactobacillus reuteri. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Lactobacillus reuteri is not identical to SEQ ID NO: 21. In some embodiments, an additional bacterial strain belonging to Lactobacillus reuteri comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 21.

[0249] In some embodiments, a composition comprises fermentates of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional bacterial strains belonging to the PCT / US25 / 16818 21 February 2025 (21.02.2025) species Lactococcus lactis. In some embodiments, the 16S rDNA sequence of an additional bacterial strain belonging to Lactococcus lactis is not identical to SEQ ID NO: 22. In some embodiments, an additional bacterial strain belonging to Lactococcus lactis comprises a 16S rDNA sequence that has at least 97%, but less than 100%, sequence identity to the nucleic acid sequence of SEQ ID NO: 22.

[0250] Some embodiments of the compositions described herein comprise fermentates of two or more bacterial strains. In some embodiments, the composition comprises fermentates of at least 2, at least 3 at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or up to 20 total bacterial strains. In some embodiments, the composition comprises fermentates of 2-20, 2-19, 2-18, 2-17, 2-16, 2-15, 2-14, 2-13, 2-12, 2-11, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-20, 3-19, 3-18, 3-17, 3-16, 3-15, 3-14, 3-13, 3-12, 3-11, 3-10, 3-9, 3-8, 3- 7, 3-6, 3-5, 3-4, 4-20, 4-19, 4-18, 4-17, 4-16, 4-15, 4-14, 4-13, 4-12, 4-11, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-20, 5-19, 5-18, 5-17, 5-16, 5-15, 5-14, 5-13, 5-12, 5-11, 5-10, 5-9, 5-8, 5-7, 5-6, 6- 20, 6-19, 6-18, 6-17, 6-16, 6-15, 6-14, 6-13, 6-12, 6-11, 6-10, 6-9, 6-8, 6-7, 7-20, 7-19, 7-18, 7-17, 7-16, 7-15, 7-14, 7-13, 7-12, 7-11, 7-10, 7-9, 7-8, 8-20, 8-19, 8-18, 8-17, 8-16, 8-15, 8- 14, 8-13, 8-12, 8-11, 8-10, 8-9, 9-20, 9-19, 9-18, 9-17, 9-16, 9-15, 9-14, 9-13, 9-12, 9-11, 9- 10, 10-20, 10-19, 10-18, 10-17, 10-16, 10-15, 10-14, 10-13, 10-12, 10-11, 11-20, 11-19, Ills, 11-17, 11-16, 11-15, 11-14, 11-13, 11-12, 12-20, 12-19, 12-18, 12-17, 12-16, 12-15, 12-

[0251] 14, 12-13, 13-20, 13-19, 13-18, 13-17, 13-16, 13-15, 13-14, 14-20, 14-19, 14-18, 14-17, 14-

[0252] 16, 14-15, 15-20, 15-19, 15-18, 15-17, 15-16, 16-20, 16-19, 16-18, 16-17, 17-20, 17-19, 17-

[0253] 18, 18-20, 18-19, or 19-20 total bacterial strains.

[0254] It will be appreciated that the terms “bacterial strains,” “microbial strains” “microbes,” “bacterial cells” and “microorganisms” are used interchangeably herein.

[0255] Bacterial strains identified as described herein were compared with sequences in publicly available nucleic acid databases, such as Basic Local Alignment Search Tool (BLAST) to determine closely related genera and species and were analyzed using taxonomic assignment tools, such as RDP Classifier, which assign bacterial taxonomy to representative sequences.

[0256] 16S ribosomal DNA (16S rDNA) sequences encoding 16S ribosomal RNA (16S rRNA) are provided below for representative bacterial strains. The closest related identified bacterial species were determined based on whole genome sequence analysis and a combination of analysis of full length 16S rDNA sequences and the sequences of single-copy conserved genes compared to publicly available sequence databases. It should be appreciated PCT / US25 / 16818 21 February 2025 (21.02.2025) that multiple bacterial strains disclosed herein may have the highest homology with the same species. In some embodiments, bacterial strains having the same or highly related 16S rDNA variable region sequences are nevertheless functionally and / or phenotypically distinct. For example, two strains may differ functionally and / or phenotypically by growth rate, antibiotic resistance, phage resistance, cell wall thickness, motility, competence, spore-forming ability, rates of metabolite (e.g., tryptophan) processing, ability to produce a given metabolite (e.g., ILA, lAld), and / or extent of production of a given metabolite. It should further be appreciated that the bacterial strains disclosed herein that have a 16S rDNA sequence with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-7, are also homologous to other strains based on their whole genome sequence, or subset of their whole genome sequence.

[0257] It should further be appreciated that the bacterial species described herein may be identified based on the nucleotide sequence of the full length 16S rDNA. Alternatively or in addition, the bacterial species described herein may be identified based on identification of 16S sequences through whole genome sequencing, and by comparing the sequences with 16S databases, or comparing the whole genome sequence, or a subset of their whole genome sequence to sequence databases.

[0258] It should be appreciated that the compositions (and / or nutritional supplements and / or food products comprising the same) may include fermentates of multiple strains of a particular species. In some embodiments, the composition includes multiple strains of a particular species that are obtained from independent sources but the strains have the same or highly related 16S rDNA sequences.

[0259] Some embodiments relate to bacterial strains with 16S rDNA sequences that have sequence identity to a nucleic acid sequence of any one of the sequences of the bacterial strains or bacterial species described herein.

[0260] “Identity” refers to the degree of sequence relatedness between or among sequences as determined by the number of matching positions between strings of two or more nucleotide sequences. “Percent (%) identity” or “percent (%) sequence identity” as it applies to nucleotide sequences is defined as the percentage of nucleotides in the candidate nucleotide sequence that are identical to the nucleotide sequence of a reference sequence after aligning the sequences and introducing gaps, as necessary, to achieve the maximum score allowed by the alignment algorithm and parameters.

[0261] Where an alignment between two sequences is contemplated, the first sequence (e.g., candidate sequence) is aligned to the second sequence (e.g., reference sequence) using the PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0262] Needleman- Wunsch algorithm for global alignment of the two sequences. Needleman & Wunsch, J Mol Biol. 1970. 48:443-453. Where two nucleotide sequences are aligned, the alignment uses an EDNAFULL substitution scoring matrix, a Gap Open penalty of 10, a Gap Extend penalty of 0.5, and no End Gap penalties. The skilled artisan will appreciate that at the time of filing the instant specification, these parameters are the default parameters of the EMBOSS Needle pairwise comparison tool provided by European Bioinformatics Institute (see ebi.ac.uk). Other suitable alignment programs may be used to obtain a global alignment using these parameters, such as BLAST, or the Needleman- Wunsch algorithm may be implemented in a scripting language (e.g., Python).

[0263] The percent sequence identity that a candidate sequence (e.g., as present in a bacterial strain encompassed by a claim) has to a reference sequence (e.g., having a SEQ ID NO. specified herein and recited in a claim) is calculated by (i) aligning the candidate sequence to the reference sequence, (ii) determining the number of matching nucleotides between the aligned candidate and reference sequences, and (iii) dividing the number of matching nucleotides by the length of the reference sequence, including any internal gaps introduced into the reference sequence when the two sequences are aligned.

[0264] The skilled artisan will appreciate that to determine whether a candidate nucleotide sequence comprises a nucleotide sequence with a given percentage sequence identity to a reference sequence, the denominator (length of reference sequence plus internal gaps) in calculating percent identity need not include gaps shown extending past the ends of the reference sequence in an alignment. Such gaps are added where a candidate sequence contains additional nucleotides that extend beyond the portions of the candidate sequence that align to the 5' end or 3' end of the reference sequence.

[0265] Some embodiments relate to bacterial strains comprising a genome (z.e., a chromosome having a nucleotide sequence) with a specified average nucleotide identity (ANI) and / or alignment fraction (AF) to a reference genome. ANI refers to average nucleotide identity of a genome (e.g., as present in a bacterial strain recited in a claim) to a reference genome (e.g., as provided in an Accession No. or SEQ ID NO.), when comparing orthologous regions between the two genomes. The skilled artisan will understand that ANI is not a calculation of percent identity obtained by alignment of two full-length genome sequences. Rather, two genomes are compared to identify regions that are conserved between both genomes, and the average nucleotide identity of all conserved regions is calculated. Konstantinidis & Tiedje, Proc Natl Acad Sci U SA. 2005. 102(7):2567-2572. Where ANI and / or AF are contemplated, these parameters are calculated using FastANI vl.34 and default PCT / US25 / 16818 21 February 2025 (21.02.2025) configuration. See Jain et al., Nat Commim. 2018. 9( 1):5114. A genome will be understood to require the following to be suitable for calculation of ANI to a reference genome: completeness as determined by detection of at least 100 single-copy BUSCO genes. See Simao et al., Bioinformatics . 2015. 31( 19):3210— 3212; Manni et al., Mol Biol Evol. 2021. 38(10):4647-4654.

[0266] The terms “identical,” or percent “identity,” in the context of two or more nucleic acids or amino acid sequences, refer to two or more sequences or subsequences that are the same. Two sequences are “substantially identical” if two sequences have a specified percentage of amino acid residues or nucleotides that are the same (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% sequence identity) over a specified region of a nucleic acid or amino acid sequence or over the entire sequence, when compared and aligned for maximum correspondence over a comparison window, or designated region as measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection. Optionally, the identity exists over a region that is at least about 50 nucleotides in length, or more preferably over a region that is 100 to 500 or 1000 or more nucleotides in length. In some embodiments, the recited degree of identity exists over the length of the 16S rRNA or 16S rDNA sequence.

[0267] In some embodiments, the bacterial strain has at least 60%, at least 70%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least

[0268] 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or up to 100% sequence identity relative to any of the bacterial strains or bacterial species described herein over a specified region (such as a region of the 16S rDNA sequences provided herein) or over the entire sequence (such as the entire 16S rDNA). It would be appreciated by one of skill in the art that the term “sequence identity” or “percent sequence identity,” in the context of two or more nucleic acid sequences or amino acid sequences, refers to a measure of similarity between two or more sequences or portion(s) thereof. In some embodiments, the identity exists over the length of the 16S rRNA or 16S rDNA sequence.

[0269] Additionally, or alternatively, two or more sequences may be assessed for the alignment between the sequences. An alignment of 100% or “total alignment” referring to two or more nucleic acids or amino acid sequences, refers to two or more sequences or subsequences that are the same. Two sequences are “substantially aligned” if two sequences have less than 100% alignment, such as a specified percentage of amino acid residues or PCT / US25 / 16818 21 February 2025 (21.02.2025) nucleotides that are the same e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical) over a specified region or over the entire sequence, when compared and aligned for maximum correspondence over a comparison window, or designated region as measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection. Optionally, the alignment exists over a region that is at least about 50 nucleotides in length, or more preferably over a region that is 100 to 500 or 1000 or more nucleotides in length.

[0270] For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. Methods of alignment of sequences for comparison, including measuring identity between sequences, are well known in the art. See, e.g., by the local homology algorithm of Smith and Waterman (1970) Adv. Appl. Math. 2:482c, by the homology alignment algorithm of Needleman and Wunsch, J. Mol. Biol. (1970) 48:443, by the search for similarity method of Pearson and Lipman. Proc. Natl. Acad. Sci. USA (1988) 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group. Madison. WI), or by manual alignment and visual inspection (see. e.g., Brent et al., Current Protocols in Molecular Biology, John Wiley & Sons, Inc. (Ringbou ed., 2003)). Two examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. (1977) 25:3389-3402; and Altschul et al., J. Mol. Biol. (1990) 215:403-410, respectively.

[0271] As will be appreciated by one of ordinary skill in the art, nomenclature regarding bacterial genus and species names may be reclassified to reflect phylogenetic relationships of microorganisms. See, e.g., Zheng et al., Inter. J. System. And Evol. Microbiol. (2020) 70(4).

[0272] Bacterial strains may be obtained from or derived from any suitable source, such as from a food source or an environmental source. As used herein, the term “derived from” in the context of bacterial strains derived from a particular source refers to obtaining a bacterial strain from the source, which may involve isolating and / or propagating cells of a bacterial strain. In some embodiments, the bacterial strains are further manipulated, such as purified and / or analyzed, prior to use. As will be evident to one of ordinary skill in the art, reference to a bacterial strain or cells of a bacterial strain that is derived from a particular source encompasses progeny cells thereof.

[0273] Bacterial strains may be derived from a fermented food or beverage, such as cultured milk and yogurt, natto, cheese, kombucha, wine, beer, cider, miso, kimchi, sauerkraut, fermented sausage, among others. Additionally, bacterial strains may be derived from PCT / US25 / 16818 21 February 2025 (21.02.2025) cultured plants or plant protein isolates, including plants or proteins isolated from cereal grains (e.g., oats), oil seeds (e.g., sunflower seeds), legumes, pulses, beans, broad beans, fava beans, peas, chickpeas, cow peas, pigeon peas, lentils, Bambara beans, vetches, and lupins.

[0274] In some embodiments, at least one bacterial strain may be purified. In some embodiments, at least one bacterial strain may be isolated. Any of the bacterial strains described herein may be isolated and / or purified, for example, from a source such as a food source (e.g., a fermented food or beverage product) or an environmental source (e.g., soil).

[0275] Compositions comprising fermentates

[0276] Provided are postbiotic compositions comprising fermentates of bacterial strains. As used herein, the term “fermentate” refers to a product obtained by fermenting a bacterial strain in an environment suitable for fermentation, such as a culture medium. A “postbiotic composition” comprises one or more fermentates, and may be in any suitable form (e.g., in a nutritional supplement, food product, and / or combined with one or more carriers). Postbiotic compositions are distinguished from probiotic compositions (containing live bacteria) at least by the inclusion of fermentates, which comprise products of fermentation, and so may exert activity separate from that of any live bacteria that may or may not be present in the postbiotic composition. In some embodiments, the fermentate comprises products produced by the bacterial strain(s) during fermentation. In some embodiments, the fermentate comprises products produced by the bacterial strain(s) during fermentation and components of the culture medium.

[0277] In some embodiments, the bacteria are propagated or manufactured using liquid fermenters, which can support the rapid growth of bacterial species. The fermenters may be, for example, stirred tank reactors or disposable wave bioreactors. Culture media such as BL media and EG media, or similar versions of these media, can be used to support the growth of the bacterial species. The bacterial fermentate can be purified and concentrated from the fermentation broth by suitable techniques, such as centrifugation and filtration. Alternatively or in addition, the bacterial fermentates can be dried and lyophilized by suitable techniques such as those known in the art.

[0278] Compositions (e.g., fermentates and / or postbiotic compositions) may comprise one or more compounds in amounts or concentrations at or above a desired threshold. Any suitable method may be used to determine the concentration of one or more compounds in a composition, such as gas chromatography -mass spectrometry (GC-MS) or liquid chromatography / mass spectrometry triple quadrupole (LC / MS-QQQ). PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0279] In some embodiments, a composition comprises 50 mg / L or more indole-3-lactic acid (ILA). In some embodiments, the concentration of ILA in the composition is at least 10 mg / L, at least 20 mg / L, at least 30 mg / L, or at least 40 mg / L. In some embodiments, the mass of ILA in the composition is 0.01 to 10 mg, 0.1 to 10 mg, 1 to 10 mg, 0.01 to 1 mg, or 0.01 to 0.1 mg. In some embodiments, the mass of ILA in the composition is 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, or 1.0 mg.

[0280] In some embodiments, a composition comprises 10 mg / L or more indole-3- carboxaldehyde (lAld). In some embodiments, the concentration of lAld in the composition is at least 1 mg / L, at least 2 mg / L, at least 4 mg / L, at least 6 mg / L, or at least 8 mg / L. In some embodiments, the mass of lAld in the composition is 0.01 to 10 mg, 0.1 to 10 mg, 1 to 10 mg, 0.01 to 1 mg, or 0.01 to 0.1 mg. In some embodiments, the mass of lAld in the composition is 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, or 1.0 mg.

[0281] In some embodiments, a composition comprises 50 mg / L or more indole-3-pyruvic acid (IPyA). In some embodiments, the concentration of IPyA in the composition is at least 10 mg / L, at least 20 mg / L, at least 30 mg / L, or at least 40 mg / L. In some embodiments, the mass of IPyA in the composition is 0.01 to 10 mg, 0.1 to 10 mg, 1 to 10 mg, 0.01 to 1 mg, or 0.01 to 0.1 mg. In some embodiments, the mass of IPyA in the composition is 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, or 1.0 mg.

[0282] In some embodiments, a composition comprises 1.5 mg / L or more indole- 3 -propionic acid (IPA). In some embodiments, the concentration of IPA in the composition is at least 0.1 mg / L, at least 0.3 mg / L, at least 0.6 mg / L, at least 0.9 mg / L, or at least 1.2 mg / L. In some embodiments, the mass of IPA in the composition is 0.0003 to 0.3 mg, 0.003 to 0.3 mg, 0.03 to 0.3 mg, 0.0003 to 0.03 mg, or 0.0003 to 0.003 mg.

[0283] In some embodiments, a postbiotic composition comprises at least 0.1% (w / w) ILA. In some embodiments, the % (w / w) ILA is 0.1-10, 0.1-5, 0.1-2, 0.1-1, or 0.1-0.5.

[0284] In some embodiments, a postbiotic composition comprises at least 0.015% (w / w) lAld. In some embodiments, the % (w / w) lAld is 0.015-1.5, 0.015-0.75, 0.015-0.30, or 0.015-0.15.

[0285] In some embodiments, a postbiotic composition comprises at least 10% w / w IPyA relative to ILA. In some embodiments, the % (w / w) of IPyA relative to ILA is 10-100, 10- 50, or 10-20.

[0286] In some embodiments, a postbiotic composition comprises at least 1% w / w IPA relative to lAld. In some embodiments, the % (w / w) of IPA relative to lAld is 1-100, 1-50, 1-20, or 1-10. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0287] In some embodiments, a postbiotic composition comprises at least 10% w / w IAA relative to ILA. In some embodiments, the % (w / w) of IAA relative to ILA is 10-100, 10-50, or 10-20.

[0288] Compositions may comprise one or more undesired compounds in amounts or concentrations below a desired threshold. Similarly, compositions may be free of detectable amounts of one or more compounds.

[0289] In some embodiments, a composition comprises less than 1 g / L tryptophan. In some embodiments, the concentration of tryptophan in the composition is 2 g / L or less, 5 g / L or less, or 10 g / L or less. In some embodiments, the composition does not comprise a detectable amount of tryptophan.

[0290] In some embodiments, the composition does not comprise a detectable amount of indole.

[0291] In some embodiments, the composition does not comprise a detectable amount of indoxyl sulfate.

[0292] In some embodiments, a composition comprises less than 1 mg / L tryptamine. In some embodiments, the concentration of tryptamine in the composition is 2 g / L or less, 5 g / L or less, or 10 g / L or less.

[0293] In some embodiments, a composition comprises less than 1 mg / L indole-3-acetamide. In some embodiments, the concentration of indole-3-acetamide in the composition is 2 g / L or less, 5 g / L or less, or 10 g / L or less.

[0294] In some embodiments, a composition comprises less than 50 mg / L indole-3-acetic acid (IAA). In some embodiments, the concentration of IAA in the composition is 100 mg / L or less, 200 mg / L or less, 300 mg / L or less, 400 mg / L or less, or 500 mg / L or less. In some embodiments, the composition does not comprise a detectable amount of IAA.

[0295] In some embodiments, a postbiotic composition comprises less than 200% w / w tryptophan relative to ILA. In some embodiments, the % (w / w) of tryptophan relative to ILA is 175% or less, 150% or less, 125% or less, 100% or less, 75% or less, 50% or less, 25% or less, or 10% or less.

[0296] In some embodiments, a postbiotic composition comprises less than 10% w / w IAA relative to ILA. In some embodiments, the % (w / w) of IAA relative to ILA is 8% or less, 6% or less, 4% or less, 2% or less, or 1% or less.

[0297] In some embodiments, a postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA. In some embodiments, the % (w / w) of tryptamine relative to ILA is 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0298] In some embodiments, a postbiotic composition comprises less than 0.5% w / w indole- 3-acetamide relative to ILA. In some embodiments, the % (w / w) of indole-3-acetamide relative to ILA is 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less.

[0299] In some embodiments, a composition acts as a substrate for production of IPA by a bacterial strain of a mammalian intestinal microbiome. In some embodiments, the mammalian intestinal microbiome is a canine intestinal microbiome. In some embodiments, a composition acts as a substrate for production of IPA by a bacterial strain belonging to the species Clostridium sporogenes. Any bacterial strain belonging to the species Clostridium sporogenes may be used to verify this activity, such as the type strain DSM 795.

[0300] A fermentate may or may not contain bacterial cells, and bacterial cells present in the fermentate may be living or dead. In some embodiments, a composition comprising a fermentate of a bacterial strain does not comprise bacterial cells of the bacterial strain. For example, a fermentate may be separated from the bacterial cells that produced the fermentate (e.g., a culture supernatant). Bacterial cells may be separated from fermentate by any suitable method, such as centrifugation or filtration.

[0301] In some embodiments, a composition comprising a fermentate of a bacterial strain also comprises bacterial cells of the bacterial strain. In some embodiments, a composition does not comprise live bacterial cells. In some embodiments, the bacterial cells are heat- killed or pasteurized.

[0302] In some embodiments, one or more fermentates are in powder form. In some embodiments, each fermentate of a composition is in powder form. In some embodiments, a composition is in powder form. Powder is a dry, bulk solid comprised of many discrete, solid, macroscopic particles, which may flow freely when the powder (or a container in which the powder is present) is shaken or tilted. The skilled artisan will appreciate that powder form fermentates may be incorporated into a composition (e.g., with a carrier and in a defined form), where the composition as a whole has a solid form that is not a powder.

[0303] Any suitable method may be used to convert a bacterial fermentate to powder form. Non-limiting examples of powder production processes include grinding and agglomeration (e.g., of solid fermentates), crystallization and spray drying (e.g., of liquid fermentates). Advantages of converting a fermentate to powder form include, without limitation, ease of incorporation into a product intended for consumption (e.g., nutritional supplement or food product intended for ingestion or dental supplement intended for chewing), distribution throughout a product to limit the fermentate mixture’s effect on taste and texture, and PCT / US25 / 16818 21 February 2025 (21.02.2025) reduction of water activity to improve stability of desired components of a fermentate mixture.

[0304] In some embodiments, a bacterial fermentate mixture in powder form has a water content of 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less, 2% or less, or 1% or less.

[0305] In some embodiments, one or more fermentates are spray-dried or lyophilized. In some embodiments, one or more fermentates are spray-dried. In some embodiments, each fermentate of a composition is spray-dried. In some embodiments, one or more fermentates are lyophilized. In some embodiments, each fermentate of a composition is lyophilized. Where a fermentate or composition is lyophilized and in powder form, powder may be lyophilized, or a lyophilized non-powder solid may be made into a powder form (e.g., by grinding and / or agglomeration).

[0306] Methods of lyophilizing compositions, specifically compositions comprising bacteria and fermentates thereof, are known in the art. See, e.g., U.S. Patent No. 3,261,761; U.S. Patent No. 4,205,132; and PCT Publication Nos. WO 2014 / 029578, WO 2012 / 098358, and WO 2019 / 226707, herein incorporated by reference to the extent they disclose lyophilization and other preservation methods. Fermentates may be lyophilized as a combination and / or the fermentates may be lyophilized separately and combined prior to incorporation into a composition, nutritional supplement, dental supplement, and / or food product. A fermentate may be combined with a nutrient or carrier prior to combining it with another fermentate or multiple lyophilized fermentates may be combined while in lyophilized form and the mixture of fermentates, once combined may subsequently be combined with a nutrient and / or excipient. In some embodiments, the fermentate or combination thereof is a lyophilized cake. In such embodiments, the fermentate(s) may be rehydrated or suspended and / or cultured prior to use. In some embodiments, the fermentate(s) in lyophilized form are used directly, for example, without rehydrating or suspension e.g., directly added to a composition).

[0307] The process of spray-drying refers to production of a dry powder from a liquid comprising a component, such as a fermentate. See, e.g., Ledet el al., Spray-Drying of Pharmaceuticals in “Lyophilized Biologies and Vaccines” pages 273-194, Springer. In general, the process involves rapidly drying the fermentates with a hot gas. A fermentate may be combined with an excipient or nutrient prior to combining it with other fermentates, or multiple spray-dried fermentates may be combined while in spray-dried form and the mixture of fermentates, once combined, may be subsequently combined with an excipient, carrier, or nutrient. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0308] The amount of fermentate in a composition is quantified according to dry weight of the fermentate. In some embodiments, a fermentate is present in a composition at an amount of 10 to 1000 mg. In some embodiments, a fermentate is present in a composition at an amount of 10 to 900 mg, 10 to 800 mg, 10 to 700 mg, 10 to 600 mg, 10 to 500 mg, 10 to 400 mg, 10 to 300 mg, 10 to 200 mg, or 10 to 100 mg. In some embodiments, a fermentate is present in a composition at an amount of 900 to 1000 mg, 800 to 1000 mg, 700 to 1000 mg, 600 to 1000 mg, 500 to 1000 mg, 400 to 1000 mg, 300 to 1000 mg, 200 to 1000 mg, or 100 mg to 1000 mg. In some embodiments, a fermentate is present in a composition in an amount of 10 to 100 mg, 100 to 200 mg, 200 to 300 mg, 300 to 400 mg, 400 to 500 mg, 500 to 600 mg, 600 to 700 mg, 700 to 800 mg, 800 to 900 mg, or 900 to 1000 mg.

[0309] In some embodiments, the total mass of fermentates in a composition is 10 to 1000 mg. In some embodiments, the total mass of fermentates in a composition is of 10 to 900 mg, 10 to 800 mg, 10 to 700 mg, 10 to 600 mg, 10 to 500 mg, 10 to 400 mg, 10 to 300 mg, 10 to 200 mg, or 10 to 100 mg. In some embodiments, the total mass of fermentates in a composition is 900 to 1000 mg, 800 to 1000 mg, 700 to 1000 mg, 600 to 1000 mg, 500 to 1000 mg, 400 to 1000 mg, 300 to 1000 mg, 200 to 1000 mg, or 100 mg to 1000 mg. In some embodiments, the total mass of fermentates in a composition is 10 to 100 mg, 100 to 200 mg, 200 to 300 mg, 300 to 400 mg, 400 to 500 mg, 500 to 600 mg, 600 to 700 mg, 700 to 800 mg, 800 to 900 mg, or 900 to 1000 mg.

[0310] In some embodiments, a fermentate is present in a composition at an amount of 0.1 to 1000 mg. In some embodiments, a fermentate is present in a composition at an amount of 0.1 to 900 mg, 0.1 to 800 mg, 0.1 to 700 mg, 0.1 to 600 mg, 0.1 to 500 mg, 0.1 to 400 mg, 0.1 to 300 mg, 0.1 to 200 mg, or 0.1 to 100 mg. In some embodiments, a fermentate is present in a composition in an amount of 0.1 to 100 mg, 100 to 200 mg, 200 to 300 mg, 300 to 400 mg, 400 to 500 mg, 500 to 600 mg, 600 to 700 mg, 700 to 800 mg, 800 to 900 mg, or 900 to 1000 mg.

[0311] In some embodiments, the total mass of fermentates in a composition is 0.2 to 2000 mg. In some embodiments, the total mass of fermentates in a composition is 0.2 to 1800 mg, 0.2 to 1600 mg, 0.2 to 1400 mg, 0.2 to 1200 mg, 0.2 to 1000 mg, 0.2 to 800 mg, 0.2 to 600 mg, 0.2 to 400 mg, or 0.2 to 200 mg. In some embodiments, the total mass of fermentates in a composition is 0.2 to 200 mg, 200 to 400 mg, 400 to 600 mg, 600 to 800 mg, 800 to 1000 mg, 1000 to 1200 mg, 1200 to 1400 mg, 1400 to 1600 mg, 1600 mg to 1800 mg.

[0312] In some embodiments, the fermentates are present in substantially equal masses in a composition. In some embodiments, the first fermentate is present in a composition in an PCT / US25 / 16818 21 February 2025 (21.02.2025) amount that is 10% to 1000% the mass of the second fermentate. In some embodiments, the second fermentate is present in a composition in an amount that is 10% to 1000% the mass of the first fermentate. In some embodiments, a composition comprises a 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1 ratio by mass of the first fermentate to the second fermentate.

[0313] Nutritional supplements and food products

[0314] Some aspects relate to a nutritional supplement comprising any of the compositions (e.g., postbiotic compositions) or fermentates described herein. Some aspects relate to a food product comprising any of the compositions (e.g., postbiotic compositions) or fermentates described herein. Food products and nutritional supplements are, in general, intended for the consumption of a human or other animal, such as a companion animal (e.g., a dog), or a bird. As will be appreciated by one of ordinary skill in the art, a food product provides a primary caloric and nutritive source intended for consumption by a subject (e.g., a feed or treat). In contrast, a nutritional supplement is not intended to provide a primary caloric or nutritive source for a subject but rather provides a targeted effect. Nutritional supplements may provide additional nutrients or functional ingredients that complement a standard diet, e.g.da standard pet diet.

[0315] Any of the bacterial strains, fermentates thereof, or combinations thereof may be present in a nutritional supplement or food product. The compositions disclosed herein can be used in a food or beverage, such as a health food, a pet food, a functional food, a dietary supplement, a food or beverage for patients, or an animal feed.

[0316] Non-limiting examples of the forms of nutritional supplements include carbohydrate- containing foods such as rice food products; paste products such as fish, hams, sausages, paste products of seafood; pouch products. Further, the examples also include food products and nutritional supplements prepared in the forms of powders, granules, kibbles, tablets, capsules, liquids, pastes, chews, and jellies.

[0317] Any suitable form may be used for a food product or nutritional supplement. Nonlimiting examples of form factors include chews with softer interiors than exteriors, kibbles, and spreads. Additional examples of form factors include a gelatinized starch matrix, extruded or injection-molded dental stick, extruded or injection-molded dental chew, chew having a soft interior, powder meal topper, water additive, dry kibble or food, wet food, frozen food, liquid spray, peanut butter spread, and soft jerky chew.

[0318] In some embodiments, the nutritional supplement is a chew having a soft interior. In some embodiments, the nutritional supplement is a kibble. In some embodiments, the kibble PCT / US25 / 16818 21 February 2025 (21.02.2025) is an extruded kibble. Extruded and / or injection-molded chews, sticks, and kibbles are known in the art, and any suitable method may be used to manufacture such chews, sticks, and kibbles. In some embodiments, the food product, or nutritional supplement is a soft jerky chew. Jerky chews may be made from any suitable starting material, such as dried meat. In some embodiments, the food product or nutritional supplement is a spread. In some embodiments, the spread comprises one or more nuts. In some embodiments, the spread is a peanut butter spread. The skilled artisan will appreciate that a food product or supplement intended for consumption by a particular animal will not contain ingredients that are toxic to that animal (e.g., macadamia nuts for dogs).

[0319] In some embodiments, the nutritional supplement is an additive. Additives may be added to other foods or food products before consumption by an animal. In some embodiments, the nutritional supplement is a water additive. Water additives are added to drinking water. In some embodiments, the nutritional supplement is a meal topper. Meal toppers may be added directly to food before consumption, with or without incorporation before consumption. In some embodiments, the meal topper is in powder form (a powder meal topper). In some embodiments, the nutritional supplement is a liquid spray. Liquid sprays may be administered directly to an animal, or sprayed on food or in water, with or without incorporation before consumption.

[0320] In some embodiments, the food product or nutritional supplement is in the form of a wet food. In some embodiments, the food product or dental supplement is in the form of a frozen food. Wet foods and frozen foods may be in any suitable form, such as those described in Niamnuy, C., & Devahastin, S. (2010). 11 Pet Foods and Their Physicochemical Properties as Affected by Processing. Contemporary Food Engineering, 327.

[0321] In some embodiments, the nutritional supplement or food product comprises a fermentate or combination thereof, and a carrier. The carrier may contain any combination of nutrients, excipients, diluents, and / or binders. In some embodiments, the carrier comprises one or more of the following compounds: water, glycerol, ethylene glycol, 1,2-propylene glycol or 1,3-propylene glycol, sodium chloride, sodium benzoate, potassium sorbate, sodium sulfate, potassium sulfate, magnesium sulfate, sodium thiosulfate, calcium carbonate, sodium citrate, dextrin, maltodextrin, glucose, sucrose, sorbitol, lactose, whey, whey permeate, wheat flour, wheat bran, com gluten meal, starch and cellulose. In some embodiments, the carrier is a gelatinized starch matrix. In some embodiments, the carrier is maltodextrin. Maltodextrin can be derived from any starch, such as rice, potato, com, wheat, or cassava. In some PCT / US25 / 16818 21 February 2025 (21.02.2025) embodiments, the carrier is a tapioca-based maltodextrin. Any suitable method may be used to obtain maltodextrin from tapioca, such as isolation from a tapioca plant.

[0322] In some embodiments, the carrier is present in a 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1 ratio by weight relative to the total amount of fermentates. In some embodiments, the carrier is present in a 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10 ratio by weight relative to the total amount of fermentates. In some embodiments, the carrier is present in a 2: 1 ratio by weight relative to the total amount of fermentates. In some embodiments, the carrier is present in a 1:1 ratio by weight relative to the total amount of fermentates. In some embodiments, the carrier is present in a 2: 1 ratio by weight relative to each fermentate. In some embodiments, the carrier is present in a 1:1 ratio by weight relative to each fermentate. In some embodiments, the carrier is present in a 1: 1 ratio by weight relative to each of three fermentates. In some embodiments, the carrier is present in a 2:1 ratio by weight relative to each of two fermentates.

[0323] In some embodiments, the carrier is present in a 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1, 37:1, 38:1, 39:1, 40:1, 41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1, or 50:1 ratio by weight, relative to the total amount of fermentates.

[0324] In some embodiments, a nutritional supplement and / or food product comprises a composition comprising fermentate(s) at an incorporation rate of 0.01% to 20% w / w. As used herein, an “incorporation rate” refers to the weight of the fermentate-containing composition as a percentage of the total weight of the nutritional supplement, or food product. For example, 10 mg of a fermentate-containing composition in a 50 g supplement is present at an incorporation rate of 0.2% w / w. In some embodiments, the incorporation rate is 0.01% to 20% w / w. In some embodiments, the incorporation rate is 0.01% to 15% w / w. In some embodiments, the incorporation rate is 0.01% to 10% w / w. In some embodiments, the incorporation rate is 0.01% to 5% w / w. In some embodiments, the incorporation rate is 0.02% to 10%, 0.03% to 10%, 0.04% to 10%, 0.05% to 10%, 0.06% to 10%, 0.07% to 10%, 0.08% to 10%, 0.09% to 10%, 0.1% to 10%, 0.2% to 10%, 0.3% to 10%, 0.4% to 10%, 0.5% to 10%, 0.6% to 10%, 0.7% to 10%, 0.8% to 10%, 0.9% to 10%, 1% to 10%, 2% to 10%, 3% to 10%, 4% to 10%, 5% to 10%, 6% to 10%, 7% to 10%, 8% to 10%, or 9% to 10% w / w. In some embodiments, the incorporation rate is 0.1% to 10%, 0.1% to 9%, 0.1% to 8%, 0.1% to 7%, 0.1% to 6%, 0.1% to 5%, 0.1% to 4%, 0.1% to 3%, 0.1% to 2%, or 0.1% to 1% w / w. In some embodiments, the incorporation rate is 0.1% to 2%, 2% to 4%, 4% to 6%, 6% to 8%, 8% to 10%, 10% to 15%, or 15% to 20% w / w. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0325] In some embodiments, a nutritional supplement and / or food product comprises a certain mass of a composition comprising fermentate(s). Defined masses of compositions are useful, for example, for supplements and / or food products that may be diluted in food or water before consumption. In some embodiments, 2 to 2,000 mg of a fermentate-containing composition is present in a nutritional supplement and / or food product. In some embodiments, 20 to 2,000, 20 to 1,800, 20 to 1,600, 20 to 1,400, 20 to 1,200, 20 to 1,000, 20 to 900, 20 to 800, 20 to 700, 20 to 600, 20 to 500, 20 to 400, 20 to 300, 20 to 200, 20 to 100, or 20 to 50 mg of the composition is present. In some embodiments, 50 to 2,000, 100 to 2,000, 200 to 2,000, 300 to 2,000, 400 to 2,000, 500 to 2,000, 600 to 2,000, 700 to 2,000, 800 to 2,000, 900 to 2,000, 1,000 to 2,000, 1,200 to 2,000, 1,400 to 2,000, 1,600 to 2,000, or 1,800 to 2,000 mg of the composition is present. In some embodiments, 100 to 200, 200 to 300, 300 to 400, 400 to 500, 500 to 600, 600 to 700, 700 to 800, 800 to 900, 1,000 to 1,200, 1,200 to 1,400, 1,400 to 1,600, 1,600 to 1,800, or 1,800 to 2,000 mg of the composition is present. In some embodiments, 0.01 to 0.1, 0.1 to 1, 1 to 10, 10 to 100, 100 to 1,000, or 1,000 to 10,000 mg of the composition is present.

[0326] In some embodiments, a nutritional supplement and / or food product is formulated for administration to subject to deliver a specified amount of one or more compounds, including ILA and lAld. A nutritional supplement and / or food product is formulated for administration to a canine subject at an ILA dose of 0.8 pg / kg per day to 8 mg / kg per day, and an lAld dose of 0.08 pg / kg per day to 800 pg / kg per day. In some embodiments, the ILA dose is 0.8 pg / kg per day to 8 pg / kg per day, 8 pg / kg per day to 80 pg / kg per day, 80 pg / kg per day to 800 pg / kg per day, or 800 pg / kg per day to 8 mg / kg per day. In some embodiments, the lAld dose is 0.08 pg / kg per day to 0.8 pg / kg per day, 0.8 pg / kg per day to 8 pg / kg per day, 8 pg / kg per day to 80 pg / kg per day, or 80 pg / kg per day to 800 pg / kg per day.

[0327] A nutritional supplement and / or food product may be formulated in a single dosage form that is administered at once, delivering an entire day’s dose in the provision of one dosage form (e.g., one treat or one meal). A nutritional supplement and / or food product may be formulated in multiple dosage forms to be administered over the course of a day, such that a certain total amount of ILA and lAld administered in a day. For example, a nutritional supplement may be formulated to be administered three times per day, with each administration providing a third of the specified dose. In some embodiments, the nutritional supplement is formulated for administration in 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 doses per day.

[0328] A nutritional supplement and / or food product may comprise one or more nutrients. The nutrient may be a carbohydrate or carbohydrate source. Non-limiting examples of PCT / US25 / 16818 21 February 2025 (21.02.2025) carbohydrate sources include grains, such as com, rice, milo, sorghum, barley, alfalfa, oats, and wheat. In some embodiments, the nutrient is a grain. In some embodiments, the nutrient is wheat, com, rice, oats, and / or barley. A nutritional supplement and / or food product may comprise a gelatinized starch matrix. Matrices of gelatinized starch may comprise any suitable starch, such as a starch listed herein or another starch known in the art.

[0329] A nutritional supplement and / or food product may comprise a fiber. Fibers may add bulk to a composition and may be fermented by intestinal bacteria to produce short-chain fatty acids and other metabolites. Non-limiting examples of fibers include beet pulp (e.g., sugar beet pulp), gum arabic, gum talha, psyllium, rice bran, carob bean gum, citrus pulp, pectin, fructooligosaccharide, mannaoligofructose, soy fiber, arabinogalactan, galactooligosaccharide, arabinoxylan, chicory pulp, chicory, coconut endosperm fiber, wheat fiber, and inulin.

[0330] A nutritional supplement and / or food product may comprise an algal strain. Algae may add one or more nutrients and / or bulk to a composition. Non-limiting examples of algal species include Ascophylum nodosum, Spirulina (e.g., Arthrospira platensis, Arthrospira fusiformis, and Arthrospira maxima), and / or Fucus vsiculosus.

[0331] A nutritional supplement and / or food product may comprise one or more inorganic minerals. Non-limiting examples of minerals include calcium carbonate, calcium, boron, selenium, calcium chloride, chloride, ferrous fumarate, zinc acetate, choline chloride, chromium, ferrous gluconate, zinc sulfate, chromium, tripicolinate, cobalt, magnesium oxide, zinc gluconate, dicalcium phosphate, copper, magnesium sulfate, ferrous sulfate, iodine, magnesium carbonate, monosodium phosphate, iron, chromium picolinate, potassium chloride, magnesium, calcium citrate, potassium citrate, manganese, calcium lactate, potassium sorbate, phosphorus, calcium gluconate, sodium bisulfate, potassium, chromium chloride, sodium hexametaphosphate, sodium, chromium nicotinate, tricalcium phosphate, zinc, and chromium citrate.

[0332] A nutritional supplement and / or food product may comprise one or more protein sources. Non-limiting examples of such protein sources include chicken meals, chicken, chicken byproduct meals, eggs, lamb, lamb meals, turkey, turkey meals, beef, beef byproducts, viscera, fish, fish meal, enterals, kangaroo, white fish, venison, blood marrow, bone marrow, soybean meal, soy protein isolate, and soy protein concentrate.

[0333] A nutritional supplement, and / or food product may comprise one or more starches. Non-limiting examples of such starches include cereals, grains, corn, wheat, rice, oats, corn grits, sorghum, grain sorghum, milo, wheat bran, oat bran, amaranth, durum, and semolina. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0334] A nutritional supplement and / or food product may comprise one or more dairy products. Dairy products include, without limitation, cream, milk, butter, and cheese.

[0335] A nutritional supplement and / or food product may comprise a meat or animal-derived material. In some embodiments, the meat or animal-derived material is beef, chicken, eggs, turkey, lamb, fish, blood marrow, bone marrow, or a combination of any thereof.

[0336] Fermentation methods

[0337] Some aspects relate to methods of preparing compositions by fermenting one or more bacterial strains to produce fermentates and combining the fermentates to prepare a postbiotic composition. It was surprisingly discovered that certain bacterial strains, while independently capable of producing several distinct indole-containing compounds (e.g., ILA and lAld), could produce greater amounts of one compound using certain fermentation conditions. By fermenting bacterial strains separately under different conditions, individual fermentates containing substantial amounts of distinct indole-containing compounds may be produced and combined to produce a postbiotic composition comprising high amounts of each indolecontaining compound.

[0338] For example, a method for producing a postbiotic composition comprises: (i) fermenting a first bacterial strain, the first bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 7, in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA; (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3- carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld, thereby producing a second fermentate comprising lAld; and (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld.

[0339] In some aspects, a method for producing a postbiotic composition comprises: (i) fermenting a first bacterial strain of Pediococcus acidlactici in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA; (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld, thereby producing a second fermentate comprising lAld; and (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0340] In some aspects, a method for producing a postbiotic composition comprises: (i) fermenting a first bacterial strain, the first bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1, in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA; (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld, thereby producing a second fermentate comprising lAld; and (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld.

[0341] In some aspects, a method for producing a postbiotic composition comprises: (i) fermenting a first bacterial strain of Lactiplantibacillus plantarum in a first fermentation medium under conditions for production of indole- 3 -lactic acid (ILA), thereby producing a first fermentate comprising ILA; (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld, thereby producing a second fermentate comprising lAld; and (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld.

[0342] In some aspects, a method for producing a postbiotic composition comprises: (i) fermenting a first bacterial strain, the first bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 7, in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA; (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld, thereby producing a second fermentate comprising lAld; and (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld.

[0343] In the course of fermentation under conditions for producing ILA or lAld, the bacterial strain may also produce one or more additional indole-containing compounds, such that the fermentate further comprises one or more indole-containing compounds in addition to ILA or lAld.

[0344] In some embodiments, the fermentation is carried out under conditions for production of both ILA and IPyA. In some embodiments, the fermentation is carried out under conditions for production of both ILA and IPA. In some embodiments, the fermentation is carried out under conditions for production of ILA, IPA, and IPyA. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0345] In some embodiments, the fermentation is carried out under conditions for production of both ILA and IA. In some embodiments, the fermentation is carried out under conditions for production of both ILA and IAA. In some embodiments, the fermentation is carried out under conditions for production of ILA, IA, and IAA.

[0346] In some embodiments, the fermentation is carried out under conditions for production of both lAld and IPyA. In some embodiments, the fermentation is carried out under conditions for production of both lAld and IPA. In some embodiments, the fermentation is carried out under conditions for production of lAld, IPA, and IPyA.

[0347] In some embodiments, the fermentation is carried out under conditions for production of both lAld and IA. In some embodiments, the fermentation is carried out under conditions for production of both lAld and IAA. In some embodiments, the fermentation is carried out under conditions for production of lAld, IA, and IAA.

[0348] Some embodiments of methods for preparing a postbiotic composition include conducting one or more additional fermentations to produce one or more additional fermentates, with such additional fermentate(s) being combined with the fermentates comprising ILA and lAld. Additional fermentations may be carried out, for instance, to separate bacterial strains that may inhibit production of one or more desired compounds by another strain when present in the same fermentation medium. Separating bacterial strains also reduces competition for resources between co-fermenting bacterial strains, avoiding the need to adjust relative amounts of different bacterial strains to achieve a desired fermentate composition.

[0349] In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 additional fermentations are carried out, with the fermentate of each additional fermentation being combined with the first and second fermentates comprising ILA and lAld. In some embodiments, one or more additional fermentations with a bacterial strain of Pediococcus acidilactici are performed. In some embodiments, one or more additional fermentations with a bacterial strain of Bacillus licheniformis are performed. In some embodiments, one or more additional fermentations with a bacterial strain of Bacillus pumilus are performed. In some embodiments, one or more additional fermentations with a bacterial strain of Bacillus subtilis are performed. In some embodiments, one or more additional fermentations with a bacterial strain of Bacillus velezensis are performed. In some embodiments, one or more additional fermentations with a bacterial strain of Weizmannia coagulans are performed. In some embodiments, one or more additional fermentations with a bacterial strain of Lactiplantibacillus plantarum are performed. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0350] In additional fermentations, a fermentation medium may comprise any combination of bacterial strains selected from Pediococcus acidilactici, Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans, and Lactiplantibacillus plantarum.

[0351] In some embodiments, one or more additional fermentations are conducted under conditions for producing IPyA. In some embodiments, one or more additional fermentations are conducted under conditions for producing IPA.

[0352] In some embodiments, one or more additional fermentations are conducted under conditions for producing IA. In some embodiments, one or more additional fermentations are conducted under conditions for producing IAA.

[0353] Fermentation under conditions for production of ILA, fermentation under conditions for production of lAld, and / or additional fermentations may be conducted under conditions such that a fermentate comprises a compound in amount at or greater than a threshold. In some embodiments, fermentation under conditions for production of ILA is carried out such that the fermentate comprises at least 50 mg / L ILA. In some embodiments, fermentation under conditions for production of lAld is carried out such that the fermentate comprises at least 10 mg / L lAld.

[0354] In some embodiments, a fermentation is carried out such that the fermentate comprises at least 50 mg / L IPyA. In some embodiments, a fermentation is carried out such that the fermentate comprises at least 1.5 mg / L IPA. In some embodiments, a fermentation is carried out such that the fermentate comprises at least 50 mg / L IAA.

[0355] Fermentation under conditions for production of ILA, fermentation under conditions for production of lAld, and / or additional fermentations may be conducted under conditions such that a fermentate comprises an undesired compound at an amount below a threshold.

[0356] In some embodiments, a fermentation is carried out such that the concentration of tryptophan in the fermentate is 1 mg / L or less. In some embodiments, a fermentation is carried out such that the concentration of IAA in the fermentate is 50 mg / L or less. In some embodiments, a fermentation is carried out such that the concentration of tryptamine in the fermentate is 1 mg / L or less. In some embodiments, a fermentation is carried out such that the concentration of indole- 3 -acetamide in the fermentate is 1 mg / L or less.

[0357] In some embodiments, a fermentation is carried out such that the fermentate comprises at least 0.1% (w / w) ILA. In some embodiments, a fermentate mixture comprises at least 0.1% (w / w) ILA. In some embodiments, a postbiotic composition comprises at least PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0358] 0.1% (w / w) ILA. In some embodiments, the % (w / w) ILA is 0.1-10, 0.1-5, 0.1-2, 0.1-1, or 0.1-0.5.

[0359] In some embodiments, a fermentation is carried out such that the fermentate comprises at least 0.015% (w / w) lAld. In some embodiments, a fermentate mixture comprises at least 0.015% (w / w) lAld. In some embodiments, a postbiotic composition comprises at least 0.015% (w / w) lAld. In some embodiments, the % (w / w) lAld is 0.015-1.5, 0.015-0.75, 0.015-0.30, or 0.015-0.15.

[0360] In some embodiments, a fermentation is carried out such that the fermentate comprises at least 10% w / w IPyA relative to ILA. In some embodiments, a fermentate mixture comprises at least 10% w / w IPyA relative to ILA. In some embodiments, a postbiotic composition comprises at least 10% w / w IPyA relative to ILA. In some embodiments, the % (w / w) of IPyA relative to ILA is 10-100, 10-50, or 10-20.

[0361] In some embodiments, a fermentation is carried out such that the fermentate comprises at least 1% w / w IPA relative to lAld. In some embodiments, a fermentate mixture comprises at least 1% w / w IPA relative to lAld. In some embodiments, a postbiotic composition comprises at least 1% w / w IPA relative to lAld. In some embodiments, the % (w / w) of IPA relative to lAld is 1-100, 1-50, 1-20, or 1-10.

[0362] In some embodiments, a fermentation is carried out such that the fermentate comprises at least 10% w / w IAA relative to ILA. In some embodiments, a fermentate mixture comprises at least 10% w / w IAA relative to ILA. In some embodiments, a postbiotic composition comprises at least 10% w / w IAA relative to ILA. In some embodiments, the % (w / w) of IAA relative to ILA is 10-100, 10-50, or 10-20.

[0363] In some embodiments, a fermentation is carried out such that the fermentate comprises less than 200% w / w tryptophan relative to ILA. In some embodiments, a fermentate mixture comprises less than 200% w / w tryptophan relative to ILA. In some embodiments, a postbiotic composition comprises less than 200% w / w tryptophan relative to ILA. In some embodiments, the % (w / w) of tryptophan relative to ILA is 175% or less, 150% or less, 125% or less, 100% or less, 75% or less, 50% or less, 25% or less, or 10% or less.

[0364] In some embodiments, a fermentation is carried out such that the fermentate comprises less than 10% w / w IAA relative to ILA. In some embodiments, a fermentate mixture comprises less than 10% w / w IAA relative to ILA. In some embodiments, a postbiotic composition comprises less than 10% w / w IAA relative to ILA. In some embodiments, the % (w / w) of IAA relative to ILA is 8% or less, 6% or less, 4% or less, 2% or less, or 1% or less. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0365] In some embodiments, a fermentation is carried out such that the fermentate comprises less than 0.5% w / w tryptamine relative to ILA. In some embodiments, a fermentate mixture comprises less than 0.5% w / w tryptamine relative to ILA. In some embodiments, a postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA. In some embodiments, the % (w / w) of tryptamine relative to ILA is 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less.

[0366] In some embodiments, a fermentation is carried out such that the fermentate comprises less than 0.5% w / w indole-3-acetamide relative to ILA. In some embodiments, a fermentate mixture comprises less than 0.5% w / w indole-3-acetamide relative to ILA. In some embodiments, a postbiotic composition comprises less than 0.5% w / w indole-3- acetamide relative to ILA. In some embodiments, the % (w / w) of indole-3 -acetamide relative to ILA is 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less.

[0367] In some embodiments, a fermentation is carried out in a fermentation medium comprising tryptophan at a concentration of 1 g / L to 12 g / L. In some embodiments, the tryptophan concentration is 1.0 g / L to 10 g / L, 1.0 to 8.0 g / L, or 1.0 g / L to 6.0 g / L. In some embodiments, the tryptophan concentration is 2.0 g / L to 12 g / L, 4.0 to 12 g / L, or 6.0 g / L to 12 g / L.

[0368] In some embodiments, a fermentation is carried out in a fermentation medium comprising tryptophan at a concentration of 1.0 g / L to 50 g / L. In some embodiments, the tryptophan concentration is 1.0 g / L to 40 g / L, 1.0 g / L to 30 g / L, 1.0 g / L to 20 g / L, or 1.0 g / L to 15 g / L. In some embodiments, the tryptophan concentration is 10 g / L to 50 g / L, 20 g / L to 50 g / L, 30 g / L to 50 g / L, or 40 g / L to 50 g / L. In some embodiments, the tryptophan concentration is about 10 g / L, about 20 g / L, about 30 g / L, about 40 g / L, or about 50 g / L.

[0369] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the concentration of glucose in the fermentation medium is less than 4 g / L. In some embodiments, the glucose concentration is 3.5 g / L or less, 3.0 g / L or less, 2.5 g / L or less, 2.0 g / L or less, 1.5 g / L or less, 1.0 g / L or less, or 0.5 g / L or less. In some embodiments, the glucose concentration is 0.1 g / L to 4.0 g / L, 0.5g / L to 4.0 g / L, 1.0 g / L to 4.0 g / L, 2.0 g / L to 4.0 g / L, or 3.0 g / L to 4.0 g / L.

[0370] Bacterial strains may be fermented in fermentation media comprising one or more carbon sources. Non-limiting examples of carbon sources include monosaccharides (e.g., glucose), disaccharides (e.g., sucrose, trehalose), oligosaccharides, polysaccharides, starches, and cellulose. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0371] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the concentration of one or more carbon sources in the fermentation medium is 4 g / L to 200 g / L. In some embodiments, the carbon source concentration is 4 g / L to 180 g / L, 4 g / L to 160 g / L, or 4 g / L to 140 g / L. In some embodiments, the carbon source concentration is 100 g / L to 200 g / L. In some embodiments, the carbon source concentration is 1 g / L to 100 g / L. In some embodiments, the carbon source concentration is 1 g / L to 90 g / L, 1 g / L to 80 g / L, 1 g / L to 70 g / L, or 1 g / L to 60 g / L. In some embodiments, the carbon source concentration is 20 g / L to 100 g / L, 30 g / L to 100 g / L, 40 g / L to 100 g / L, or 50 g / L to 100 g / L. In some embodiments, the carbon source concentration is 1 g / L to 25 g / L, 25 g / L to 50 g / L, 50 g / L to 75 g / L, or 75 g / L to 100 g / L. In some embodiments, the carbon source is a monosaccharide. In some embodiments, the carbon source is a disaccharide. In some embodiments, the carbon source is a starch. In some embodiments, the carbon source is cellulose.

[0372] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the concentration of glucose in the fermentation medium is 4 g / L to 200 g / L. In some embodiments, the glucose concentration is 4 g / L to 180 g / L, 4 g / L to 160 g / L, or 4 g / L to 140 g / L. In some embodiments, the glucose concentration is 100 g / L to 200 g / L. In some embodiments, the glucose concentration is 1 g / L to 100 g / L. In some embodiments, the glucose concentration is 1 g / L to 90 g / L, 1 g / L to 80 g / L, 1 g / L to 70 g / L, or 1 g / L to 60 g / L. In some embodiments, the glucose concentration is 20 g / L to 100 g / L, 30 g / L to 100 g / L, 40 g / L to 100 g / L, or 50 g / L to 100 g / L. In some embodiments, the glucose concentration is 1 g / L to 25 g / L, 25 g / L to 50 g / L, 50 g / L to 75 g / L, or 75 g / L to 100 g / L.

[0373] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the concentration of glucose in the fermentation medium is at least 4.0 g / L. In some embodiments, the glucose concentration is at least 5.0 g / L, at least 6.0 g / L, at least 8.0 g / L, at least 10 g / L, at least 12 g / L, or at least 14 g / L. In some embodiments, the glucose concentration is 4.0 to 15 g / L, 4.0 g / L to 12 g / L, 4.0 g / L to 10 g / L, 4.0 g / L to 8 g / L, or 4.0 to 6.0 g / L. In some embodiments, the glucose concentration is 6.0 to 15 g / L, 8.0 g / L to 15g / L, 10 g / L to 15 g / L, 12 g / L to 15 g / L, or 14 to 15 g / L.

[0374] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the concentration of glucose in the fermentation medium is 4 g / L to 200 g / L. In some embodiments, the glucose concentration is 4 g / L to 180 g / L, 4 g / L to 160 g / L, or 4 g / L to 140 g / L. In some embodiments, the glucose concentration is 100 g / L to 200 g / L. In some embodiments, the glucose concentration is 100 g / L to 180 g / L, 100 g / L to 160 g / L, 100 PCT / US25 / 16818 21 February 2025 (21.02.2025) g / L to 140 g / L, or 100 g / L to 120 g / L. In some embodiments, the glucose concentration is 120 g / L to 200 g / L, 140 g / L to 200 g / L, 160 g / L to 200 g / L, or 180 g / L to 200 g / L. In some embodiments, the glucose concentration is 100 g / L to 120 g / L, 120 g / L to 140 g / L, 140 g / L to 160 g / L, 160 g / L to 180 g / L, or 180 g / L to 200 g / L.

[0375] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the concentration of one or more carbon sources in the fermentation medium is 4 g / L to 200 g / L. In some embodiments, the carbon source concentration is 4 g / L to 180 g / L, 4 g / L to 160 g / L, or 4 g / L to 140 g / L. In some embodiments, the carbon source concentration is 100 g / L to 200 g / L. In some embodiments, the carbon source concentration is 100 g / L to 180 g / L, 100 g / L to 160 g / L, 100 g / L to 140 g / L, or 100 g / L to 120 g / L. In some embodiments, the carbon source concentration is 120 g / L to 200 g / L, 140 g / L to 200 g / L, 160 g / L to 200 g / L, or 180 g / L to 200 g / L. In some embodiments, the carbon source concentration is 100 g / L to 120 g / L, 120 g / L to 140 g / L, 140 g / L to 160 g / L, 160 g / L to 180 g / L, or 180 g / L to 200 g / L. In some embodiments, the carbon source is a monosaccharide. In some embodiments, the carbon source is a disaccharide. In some embodiments, the carbon source is a starch. In some embodiments, the carbon source is cellulose.

[0376] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the fermentation medium comprises polysorbate 80. In some embodiments, the polysorbate 80 concentration in the fermentation medium is at least 0.1 g / L, at least 0.2 g / L, at least 0.4 g / L, at least 0.6 g / L, at least 0.8 g / L, or at least 1 g / L.

[0377] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the fermentation medium comprises manganese. In some embodiments, the manganese concentration in the fermentation medium is at least 0.01 mg / L, at least 0.02 mg / L, at least 0.04 mg / L, at least 0.06 mg / L, at least 0.08 mg / L, or at least 0.1 mg / L. In some embodiments, the manganese concentration in the fermentation medium is at least 0.01 g / L, at least 0.1 g / L, at least 1 g / L, or at least 10 g / L. In some embodiments, the manganese concentration in the fermentation medium is 0.001 g / L to 0.01 g / L, 0.01 g / L to 0.1 g / L, 0.1 g / L to 1 g / L, or 1 g / L to 10 g / L.

[0378] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the fermentation medium comprises vitamin B12. In some embodiments, the vitamin B12 concentration in the fermentation medium is at least 0.0001 mg / L, at least 0.0002 mg / L, at least 0.0004 mg / L, at least 0.0006 mg / L, at least 0.0008 mg / L, or at least 0.001 mg / L. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0379] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium does not comprise polysorbate 80. In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium does not comprise manganese. In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium does not comprise vitamin B12.

[0380] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium comprises polysorbate 80. In some embodiments, the polysorbate 80 concentration in the fermentation medium is at least 0.1 g / L, at least 0.2 g / L, at least 0.4 g / L, at least 0.6 g / L, at least 0.8 g / L, or at least 1 g / L.

[0381] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium comprises manganese. In some embodiments, the manganese concentration in the fermentation medium is at least 0.01 mg / L, at least 0.02 mg / L, at least 0.04 mg / L, at least 0.06 mg / L, at least 0.08 mg / L, or at least 0.1 mg / L. In some embodiments, the manganese concentration in the fermentation medium is at least 0.01 g / L, at least 0.1 g / L, at least 1 g / L, or at least 10 g / L. In some embodiments, the manganese concentration in the fermentation medium is 0.001 g / L to 0.01 g / L, 0.01 g / L to 0.1 g / L, 0.1 g / L to 1 g / L, or 1 g / L to 10 g / L.

[0382] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium comprises vitamin B12. In some embodiments, the vitamin B12 concentration in the fermentation medium is at least 0.0001 mg / L, at least 0.0002 mg / L, at least 0.0004 mg / L, at least 0.0006 mg / L, at least 0.0008 mg / L, or at least 0.001 mg / L.

[0383] Bacterial strains may be fermented in fermentation media comprising one or more nitrogen sources. Examples of nitrogen sources include, without limitation, yeast extract, peptones, ammonium salts, nitrates, and amino acids.

[0384] In some embodiments, a bacterial strain is fermented under conditions for ILA production, where the fermentation medium comprises 10 g / L to 125 g / L of one or more nitrogen sources. In some embodiments, the fermentation medium comprises 10 g / L to 120 g / L, 10 g / L to 100 g / L, 10 g / L to 80 g / L, 10 g / L to 60 g / L, or 10 g / L to 40 g / L of one or more nitrogen sources. In some embodiments, the fermentation medium comprises 25 g / L to 125 g / L, 50 g / L to 125 g / L, 75 g / L to 125 g / L, or 100 g / L to 125 g / L of one or more nitrogen sources. In some embodiments, the fermentation medium comprises 10 g / L to 25 g / L, 25 g / L to 50 g / L, 50 g / L to 75 g / L, 75 g / L to 100 g / L, or 100 g / L to 125 g / L of one or more nitrogen PCT / US25 / 16818 21 February 2025 (21.02.2025) sources. In some embodiments, nitrogen sources in the fermentation medium comprise yeast extract and peptones. In some embodiments, the fermentation medium does not comprise nitrogen sources other than yeast extract, peptones, or the combination of yeast extract and peptones.

[0385] In some embodiments, a bacterial strain is fermented under conditions for lAld production, where the fermentation medium comprises 125 g / L to 200 g / L of one or more nitrogen sources. In some embodiments, the fermentation medium comprises 125 g / L to 190 g / L, 125 g / L to 180 g / L, 125 g / L to 170 g / L, 125 g / L to 160 g / L, or 125 g / L to 150 g / L of one or more nitrogen sources. In some embodiments, the fermentation medium comprises 130 g / L to 200 g / L, 140 g / L to 200 g / L, 150 g / L to 200 g / L, 160 g / L to 200 g / L, or 170 g / L to 200 g / L of one or more nitrogen sources. In some embodiments, the fermentation medium comprises 125 g / L to 140 g / L, 140 g / L to 160 g / L, 160 g / L to 180 g / L, or 180 g / L to 200 g / L of one or more nitrogen sources. In some embodiments, nitrogen sources in the fermentation medium comprise yeast extract and peptones. In some embodiments, the fermentation medium does not comprise nitrogen sources other than yeast extract, peptones, or the combination of yeast extract and peptones.

[0386] In some embodiments, a bacterial fermentate mixture comprises a first fermentate and second fermentate, wherein:

[0387] (i) the first fermentate is produced by incubating a bacterial strain under microaerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12, and the second fermentate is produced by incubating a bacterial strain under aerobic conditions in a second fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin Bl 2; or

[0388] (ii) the first fermentate is produced by incubating a bacterial strain under aerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12, and the second fermentate is produced by incubating a bacterial strain in a second fermentation medium comprising a second fermentation medium comprising 1 g / L to 50 g / L tryptophan, less than 4 g / L glucose, no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

[0389] In some embodiments, a bacterial strain is fermented under the following conditions (Condition Al): a microaerobic environment; PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0390] 1 g / L to 50 g / L tryptophan;

[0391] 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12.

[0392] In some embodiments, a bacterial strain is fermented under the following conditions (Condition B): an aerobic environment;

[0393] 1 g / L to 50 g / L tryptophan;

[0394] 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12.

[0395] In some embodiments, a bacterial fermentate mixture comprises a first fermentate produced by fermentation under Condition Al and a second fermentate produced by fermentation under Condition B.

[0396] In some embodiments, a bacterial strain is fermented under the following conditions (Condition A2): an aerobic environment;

[0397] 1 g / L to 50 g / L tryptophan;

[0398] 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12.

[0399] In some embodiments, a bacterial strain is fermented under the following conditions (Condition C): an aerobic environment;

[0400] 1 g / L to 50 g / L tryptophan; less than 4 g / L glucose; and no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

[0401] In some embodiments, a bacterial fermentate mixture comprises a first fermentate produced by fermentation under Condition A2 and a second fermentate produced by fermentation under Condition C.

[0402] In some embodiments, a bacterial fermentate mixture comprises a first fermentate and second fermentate, wherein (i) the first fermentate is produced by incubating a bacterial strain under microaerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan, 100 g / L to 200 g / L glucose, the presence of manganese, and 10 g / L to 125 g / L of one or more nitrogen sources; and (ii) the second fermentate is produced by incubating a bacterial strain under aerobic conditions in a second fermentation medium comprising 1 g / L PCT / US25 / 16818 21 February 2025 (21.02.2025) to 50 g / L tryptophan, 1 g / L to 100 g / L glucose, the presence of manganese, and 125 g / L to 200 g / L of one or more nitrogen sources.

[0403] In some embodiments, a bacterial strain is fermented under the following conditions (Condition D): a microaerobic environment;

[0404] 1 g / L to 50 g / L tryptophan;

[0405] 100 g / L to 200 g / L glucose; the presence of manganese; and

[0406] 10 g / L to 125 g / L of one or more nitrogen sources.

[0407] In some embodiments, a bacterial strain is fermented under the following conditions (Condition E): an aerobic environment;

[0408] 1 g / L to 50 g / L tryptophan;

[0409] 1 g / L to 100 g / L glucose; the presence of manganese; and

[0410] 125 g / L to 200 g / L of one or more nitrogen sources.

[0411] In some embodiments, a bacterial fermentate mixture comprises a first fermentate produced by fermentation under Condition D and a second fermentate produced by fermentation under Condition E.

[0412] In some embodiments, a bacterial strain is fermented for at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least

[0413] 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least

[0414] 20, or at least 21 days. In some embodiments, a bacterial strain is fermented for up to 1, up to 2, up to 3, up to 4, up to 5, up to 6, up to 7, up to 8, up to 9, up to 10, up to 11, up to 12, up to

[0415] 13, up to 14, up to 15, up to 16, up to 17, up to 18, up to 19, up to 20, or up to 21 days. In some embodiments, a bacterial strain is fermented for 1-21, 1-20, 10-17, 1-14, 1-11, 1-10, 1- 7, 1-6, 1-5, 1-4, 1-3, or 1-2 days. In some embodiments, a bacterial strain is fermented for 2-

[0416] 21, 2-20, 10-17, 2-14, 2-11, 2-10, 2-7, 2-6, 2-5, 2-4, or 2-3 days. In some embodiments, a bacterial strain is fermented for 1-7, 2-7, 3-7, 4-7, 5-7, or 6-7 days.

[0417] In some embodiments, a bacterial strain is fermented under aerobic conditions. An environment having “aerobic conditions” contains oxygen at a concentration similar to that of Earth’s atmosphere (z.e., air is 21% O2). In some embodiments, the bacterial strain is fermented in an environment having an oxygen concentration of 17% to 25%, 18% to 24%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0418] 19% to 23%, or 20% to 22%. In some embodiments, the oxygen concentration is at least 20%.

[0419] In some embodiments, a bacterial strain is fermented under microaerobic conditions. An environment having “microaerobic conditions” contains limited amounts of oxygen, at a concentration lower than that of Earth’s atmosphere (e.g., air is 1-2% O2). In some embodiments, the bacterial strain is fermented in an environment having an oxygen concentration of 0.5% to 5%, 1.0% to 4%, or 1.5% to 3%. In some embodiments, the oxygen concentration is about 0.5%, about 1.0%, about 1.5%, or about 2.0%. In some embodiments, the oxygen concentration is 1.0% to 2.0%.

[0420] The amount of dissolved oxygen in a fermentation medium may be modulated by introducing air directly into the fermentation medium, such as by air sparging. In some embodiments, aerobic conditions are achieved by air sparging during fermentation. In some embodiments, microaerobic conditions are achieved by fermentation without air sparging. Air sparging in a fermentation bioreactor involves introducing pressurized air or oxygen through diffusers or spargers directly into the liquid medium to enhance oxygen distribution throughout the culture.

[0421] In some embodiments, the first and / or second fermentates are concentrated before being combined to form a fermentate mixture. Concentrating a fermentate refers to the removal of water from the fermentate, thereby increasing the concentration of fermentation products, such as ILA and lAld, that are present in the fermentate. A fermentate may be concentrated to reduce water content without reducing water content to the same extent as a drying process. As one example, a fermentation medium may be subjected to circulating air or reduced atmospheric pressure to facilitate evaporation.

[0422] In some embodiments, the first and / or second fermentates are dried before being combined to form a fermentate mixture. Any suitable drying method may be used to dry fermentates, including lyophilization, spray-drying, and vacuum drying. In some embodiments, one or more fermentates is dried by a process that does not raise the fermentate temperature above 100 °C, above 90 °C, above 80 °C above 70 °C, above 60 °C, or above 50 °C.

[0423] Methods of use

[0424] Some aspects relate to administration of a composition (and / or nutritional supplement and / or food product comprising the same) to a subject in need thereof. Administration may be accomplished through any suitable method, such as one described herein or known in the art, PCT / US25 / 16818 21 February 2025 (21.02.2025) and may include, for example, consumption of the composition (and / or nutritional supplement and / or food product) by the subject or topical application of the composition onto the subject. Administration may be for the treatment or prevention of a condition or one or more symptoms of the condition. “Treating” a condition refers to reducing the frequency, duration, and / or severity of one or more symptoms of the condition. “Preventing” a condition encompasses prophylactic administration and refers to reducing the likelihood of developing the condition or one or more of its symptoms.

[0425] Some aspects relate to methods of treating or preventing skin inflammation, skin irritation, and / or skin damage in a subject in need thereof. Non-limiting examples of skin inflammation symptoms include redness, swelling, itchiness, warmth, heat, discomfort, and pain. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced. In some embodiments, the symptom is a symptom of skin inflammation. In some embodiments, the symptom is a symptom of skin irritation. In some embodiments, the symptom is a symptom of photodamage. In some embodiments, the symptom is a symptom of UV damage.

[0426] Some aspects relate to treating or preventing gut inflammation or gastrointestinal distress in a subject in need thereof. Non-limiting examples of symptoms of gastrointestinal distress include bloating, constipation, diarrhea, heartbum, nausea, reflux, stomach pain and cramping, and vomiting. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0427] Some aspects relate to treating or preventing neuroinflammation in a subject in need thereof. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced. In some embodiments, mobility of the subject is improved. In some embodiments, cognition of the subject is improved.

[0428] Some aspects relate to methods of treating or preventing weight-related inflammation in a subject in need thereof. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0429] Some aspects relate to methods of limiting weight gain in a subject in need thereof. In some embodiments, the rate of weight gain is reduced. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0430] Some aspects relate to methods of promoting weight loss in a subject in need thereof. In some embodiments, the composition, nutritional supplement, and / or food product the rate of weight loss is increased.

[0431] Some aspects relate to methods of treating or preventing overweight status or obesity in a subject in need thereof. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0432] Some aspects relate to methods of treating or preventing anxiety in a subject. Nonlimiting examples of anxiety symptoms include excessive barking, pacing, trembling, aggressive behavior, destructive behavior, or withdrawal. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0433] Some aspects relate to methods of treating or preventing a seasonal allergy in a subject in need thereof. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0434] Some aspects relate to methods of treating or preventing a food allergy in a subject in need thereof. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0435] Some aspects relate to methods of treating or preventing gum inflammation in a subject in need thereof. Non-limiting examples of symptoms of gum inflammation include gum redness, swelling, pain, and bleeding. In some embodiments, the duration of one or more symptoms is reduced. In some embodiments, the frequency of one or more symptoms is reduced. In some embodiments, the severity of one or more symptoms is reduced.

[0436] In some embodiments, administration is oral. Oral administration may refer to manual administration, such as oral gavage or spraying (e.g., via aerosol). Oral administration may also refer to feeding, such as providing the composition or nutritional supplement to a subject (e.g., a companion animal), which ingests it by eating or drinking. Methods may include a single administration or administration may be repeated. Repeat administrations may be conducted at regular intervals (e.g., daily or weekly). For example, a subject may be administered a composition or supplement one or more times daily, over a specified time PCT / US25 / 16818 21 February 2025 (21.02.2025) period, such as a week, several weeks, a month, or several months. Alternatively, multiple instances of administration may be conducted on an as-needed basis.

[0437] In some embodiments, administration is rectal. In some embodiments, a composition is in the form of a suppository when administered rectally. In some embodiments, a composition, supplement, or food product is delivered to the intestine. In some embodiments, a composition, supplement, or food product is delivered to the colon. Such delivery to the intestine or colon may be accomplished by oral or rectal administration.

[0438] In some embodiments, administration is topical. Topical administration refers to application of a composition to a particular body surface, such as the skin or mucus membranes. Forms of compositions suitable for topical administration include, without limitation, ointments, gels, creams, foams, emulsions, lotions, pastes, powders, tinctures, drops, rinses, sprays, and transdermal patches. These and other delivery vehicles for topical administration of compositions are known in the art. See, e.g., Mayba and Gooderham, J Cutan Med Surg. 2018. 22(2):207-212. In some embodiments, a composition is applied to the skin by topical administration.

[0439] In some embodiments, a composition comprising fermentate(s) is present in a topical administration formulation (z.e., a composition in a form for topical application) at an incorporation rate of 0.001% to 10% w / w. As used in this paragraph, an “incorporation rate” refers to the mass of the fermentate-containing composition as a percentage of the total weight of the topical administration formulation. For example, 10 mg of a fermentate- containing composition in a 50 g topical administration formulation is present at an incorporation rate of 0.2% w / w. In some embodiments, the incorporation rate is 0.001% to 10% w / w. In some embodiments, the incorporation rate is 0.001% to 5% w / w. In some embodiments, the incorporation rate is 0.001% to 1% w / w. In some embodiments, the incorporation rate is 0.001% to 0.1% w / w. In some embodiments, the incorporation rate is 0.001% to 0.01% w / w. In some embodiments, the incorporation rate is 0.002% to 10%, 0.003% to 10%, 0.004% to 10%, 0.005% to 10%, 0.006% to 10%, 0.007% to 10%, 0.008% to 10%, 0.009% to 10%, 0.01% to 10%, 0.1% to 10%, or 1% to 10% w / w. In some embodiments, the incorporation rate is 0.001% to 0.003%, 0.003% to 0.01%, 0.01% to 0.03%, 0.03% to 0.1%, 0.1% to 0.3%, 0.3% to 1%, 1% to 3%, or 3% to 10% w / w.

[0440] In some embodiments, a composition, nutritional supplement, or food product is administered in an effective amount for treating, preventing, or alleviating one or more symptoms in a subject. An “effective amount” or “effective dosage” refers to an amount or dosage, respectively, sufficient to treat, alleviate, or prevent a sign or symptom in a subject. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0441] The actual effective amount or dosage will depend on the individual subject and their health. Such effective amounts or dosages may be determined by routine assays known to those of skill in the art.

[0442] Subjects to which a composition, nutritional supplement, or food product may be administered include any animal known in the art. In some embodiments, the subject is a human. In some embodiments, the animal is a domesticated animal. In some embodiments, the animal is a carnivore. In some embodiments, the animal is a rodent. In some embodiments, the rodent is a mouse, rat, guinea pig, chinchilla, or hamster. In some embodiments, the animal is a dog, cat, rabbit, guinea pig, hamster, or ferret. In some embodiments, the animal is a dog. In some embodiments, the animal is a cat. In some embodiments, the animal is a bovine, swine, llama, alpaca, sheep, or goat. In some embodiments, the animal is a bird.

[0443] Some aspects relate to methods of treating, preventing, or reducing the extent of inflammation in a subject in need thereof. In some embodiments, the composition, nutritional supplement and / or food product is effective in reducing one or more symptoms, such as inflammation, diarrhea, vomiting, itching, and allergy. In some embodiments, the composition, nutritional supplement and / or food product reduces inflammation in the subject. In some embodiments, the composition, nutritional supplement and / or food product treats or prevents diarrhea in the subject. In some embodiments, the composition, nutritional supplement and / or food product treats or prevents vomiting in the subject. In some embodiments, the composition, nutritional supplement, and / or food product treats or prevents itching in the subject. In some embodiments, the composition, nutritional supplement, and / or food product treats or prevents allergy in the subject.

[0444] In some embodiments, the composition, nutritional supplement, and / or food product reduces inflammation in the subject. In some embodiments, the composition, nutritional supplement, and / or food product treats or prevents one or more symptoms associated with inflammation. Methods of measuring inflammation are known in the art, and include, without limitation, measuring the concentration of one or more cytokines (e.g., TNF-a, IL-ip, IL-6, IL- 10, and IL- 12) in the subject, such as in a sample obtained from the subject (e.g., a blood sample, serum sample, plasma sample). In some embodiments, the composition, nutritional supplement, and / or food product reduces infiltration of leukocytes (e.g., neutrophils and / or monocytes) into oral mucosa, gut mucosa, or dermis.

[0445] In some embodiments, administration results in a decrease in the serum concentration of one or more inflammatory cytokines by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0446] 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the serum concentration of the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the serum concentration of one or more inflammatory cytokines by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the serum concentration in a subject (e.g., a reference subject) who did not receive the composition.

[0447] In some embodiments, administration results in a decrease in the duration of inflammation by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration of inflammation in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in duration of inflammation by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to duration of inflammation in a subject (e.g., a reference subject) who did not receive the composition.

[0448] In some embodiments, administration results in a decrease in the frequency of diarrhea by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of diarrhea in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the frequency of diarrhea by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of diarrhea in a subject (e.g., a reference subject) who did not receive the composition.

[0449] In some embodiments, administration results in a decrease in the duration over which diarrhea occurs by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration over which diarrhea occurs in the same subject prior to administering the PCT / US25 / 16818 21 February 2025 (21.02.2025) composition. In some embodiments, administration results in a decrease in the duration over which diarrhea occurs by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration over which diarrhea occurs in a subject (e.g., a reference subject) who did not receive the composition.

[0450] In some embodiments, administration results in a decrease in the frequency of vomiting by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of vomiting in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the frequency of vomiting by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of vomiting in a subject (e.g., a reference subject) who did not receive the composition.

[0451] In some embodiments, administration results in a decrease in the duration over which vomiting occurs by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration over which vomiting occurs in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the duration over which vomiting occurs by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration over which vomiting occurs in a subject (e.g., a reference subject) who did not receive the composition.

[0452] In some embodiments, administration results in a decrease in the frequency of itching by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of itching in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the frequency of itching by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0453] 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of itching in a subject (e.g., a reference subject) who did not receive the composition.

[0454] In some embodiments, administration results in a decrease in the duration over which itching occurs by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration over which itching occurs in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the duration over which itching occurs by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration over which itching occurs in a subject (e.g., a reference subject) who did not receive the composition.

[0455] In some embodiments, administration results in a decrease in the frequency of allergy symptoms by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of allergy symptoms in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the frequency of allergy symptoms by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the frequency of allergy symptoms in a subject (e.g., a reference subject) who did not receive the composition.

[0456] In some embodiments, administration results in a decrease in the duration of allergy symptoms by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration of allergy symptoms in the same subject prior to administering the composition. In some embodiments, administration results in a decrease in the duration of allergy symptoms by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, as compared to the duration of allergy symptoms in a subject (e.g., a reference subject) who did not receive the composition. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0457] Reduction of inflammation and / or symptoms thereof

[0458] Fermentates of bacterial strains for use in the compositions described herein were identified based on the ability to produce indole-containing compounds that interact with aryl hydrocarbon receptors (AhRs), such as the canine AhR. Without wishing to be bound by theory, the presence of such fermentates is believed to be useful for improving homeostasis (e.g., by modulating inflammation and / or improving tissue barrier integrity).

[0459] In some embodiments, a composition, nutritional supplement, and / or food product reduces inflammation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of inflammation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents diarrhea in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents vomiting in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents itching in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of allergy in the subject.

[0460] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of skin inflammation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of skin irritation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of skin photodamage in the subject.

[0461] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of gut inflammation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of gastrointestinal distress in the subject.

[0462] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of neuroinflammation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product improves mobility in the subject. In some embodiments, a composition, nutritional supplement, and / or food product improves cognition in the subject.

[0463] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of weight-related inflammation in the subject. In some embodiments, a composition, nutritional supplement, and / or food product reduces the rate of weight gain in the subject. In some embodiments, a composition, nutritional supplement, and / or food product increases the rate of weight loss in the subject. In some embodiments, a PCT / US25 / 16818 21 February 2025 (21.02.2025) composition, nutritional supplement, and / or food product treats or prevents a symptom of overweight or obese status in the subject.

[0464] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of anxiety in the subject.

[0465] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of seasonal allergy in the subject.

[0466] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of food allergy in the subject.

[0467] In some embodiments, a composition, nutritional supplement, and / or food product treats or prevents a symptom of gum inflammation in the subject.

[0468] In some embodiments, a composition, nutritional supplement, and / or food product reduces the serum concentration of one or more inflammatory cytokines in the subject. In some embodiments, the serum cytokine concentration is reduced by at least 1.1 -fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10- fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the serum cytokine concentration in the same subject prior to prior to exposure to the composition or fermentate(s). In some embodiments, the serum cytokine concentration is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the serum cytokine concentration in the same subject prior to exposure to the composition or fermentate(s).

[0469] In some embodiments, a composition, nutritional supplement, and / or food product reduces the duration of inflammation in the subject. In some embodiments, the duration of inflammation is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3- fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the duration of inflammation in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the duration of inflammation is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the duration of inflammation in the same subject prior to exposure to the composition or fermentate(s). PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0470] In some embodiments, a composition, nutritional supplement, and / or food product reduces the frequency of one or more symptoms of inflammation in the subject. In some embodiments, the inflammation symptom frequency is reduced by at least 1.1 -fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10- fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the inflammation symptom frequency in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the inflammation symptom frequency is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the inflammation symptom frequency in the same subject prior to exposure to the composition or fermentate(s).

[0471] In some embodiments, a composition, nutritional supplement, and / or food product reduces the duration of one or more symptoms of inflammation in the subject. In some embodiments, the inflammation symptom duration is reduced by at least 1.1 -fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10- fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the inflammation symptom duration in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the inflammation symptom duration is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the inflammation symptom duration in the same subject prior to exposure to the composition or fermentate(s).

[0472] In some embodiments, a composition, nutritional supplement, and / or food product reduces the frequency of diarrhea in the subject. In some embodiments, the frequency of diarrhea is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4- fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100- fold, 1000-fold, 104-fold, 105-fold or more as compared to the frequency of diarrhea in the subject prior to exposure to the composition or fermentate(s). In some embodiments, the frequency of diarrhea is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or PCT / US25 / 16818 21 February 2025 (21.02.2025) more as compared to the frequency of diarrhea in the same subject prior to exposure to the composition or fermentate(s).

[0473] In some embodiments, a composition, nutritional supplement, and / or food product reduces the duration over which diarrhea occurs in the subject. In some embodiments, the duration over which diarrhea occurs is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4- fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30- fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the duration over which diarrhea occurs in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the duration over which diarrhea occurs is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the duration over which diarrhea occurs in the same subject prior to exposure to the composition or fermentate(s).

[0474] In some embodiments, a composition, nutritional supplement, and / or food product reduces the frequency of vomiting in the subject. In some embodiments, the frequency of vomiting is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100- fold, 1000-fold, 104-fold, 105-fold or more as compared to the frequency of vomiting in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the frequency of vomiting is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the frequency of vomiting in the same subject prior to exposure to the composition or fermentate(s).

[0475] In some embodiments, a composition, nutritional supplement, and / or food product reduces the duration over which vomiting occurs in the subject. In some embodiments, the duration over which vomiting occurs is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4- fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30- fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the duration over which vomiting occurs in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the duration over which vomiting occurs is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0476] 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the duration over which vomiting occurs in the same subject prior to exposure to the composition or fermentate(s).

[0477] In some embodiments, a composition, nutritional supplement, and / or food product reduces the frequency of itching in the subject. In some embodiments, the frequency of itching is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4- fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100- fold, 1000-fold, 104-fold, 105-fold or more as compared to the frequency of itching in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the frequency of itching is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the frequency of itching in the same subject prior to exposure to the composition or fermentate(s).

[0478] In some embodiments, a composition, nutritional supplement, and / or food product reduces the duration over which itching occurs in the subject. In some embodiments, the duration over which itching occurs is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to the duration over which itching occurs in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, the duration over which itching occurs is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to the duration over which itching occurs prior to prior to exposure to the composition or fermentate(s).

[0479] In some embodiments, a composition, nutritional supplement, and / or food product reduces the frequency of one or more symptoms of allergy in the subject. In some embodiments, the allergy symptom frequency is reduced by at least 1.1-fold, 1.2-fold, 1.3- fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to allergy symptom frequency in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, allergy symptom frequency is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0480] 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to allergy symptom frequency in the same subject prior to exposure to the composition or fermentate(s).

[0481] In some embodiments, a composition, nutritional supplement, and / or food product reduces the duration of one or more symptoms of allergy in the subject. In some embodiments, the allergy symptom duration is reduced by at least 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, 104-fold, 105-fold or more as compared to allergy symptom duration in the same subject prior to exposure to the composition or fermentate(s). In some embodiments, allergy symptom duration is reduced by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more as compared to allergy symptom duration in the same subject prior to exposure to the composition or fermentate(s).

[0482] ENUMERATED EMBODIMENTS

[0483] 1. A postbiotic composition comprising a first fermentate and a second fermentate of a bacterial strain, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 (Pediococcus acidilaclici) or SEQ ID NO: 7 (Lactiplantibacillus plantarum), and wherein the first fermentate comprises indole-3 -lactic acid (ILA) and the second fermentate comprises indole-3-carboxaldehyde (lAld).

[0484] 2. The postbiotic composition of Embodiment 1, wherein the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1.

[0485] 3. The postbiotic composition of Embodiment 1 or 2, wherein the bacterial strain belongs to the species Pediococcus acidilactici.

[0486] 4. The postbiotic composition of any one of Embodiments 1-3, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0487] 5. The postbiotic composition of any one of Embodiments 1-4, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 50.0% to the reference genome of SEQ ID NO: 8.

[0488] 6. The postbiotic composition of any one of Embodiments 1-5, wherein the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising:

[0489] 1 g / L to 50 g / L tryptophan;

[0490] 100 g / L to 200 g / L glucose; manganese; and

[0491] 10 g / L to 125 g / L of one or more nitrogen sources.

[0492] 7. The postbiotic composition of any one of Embodiments 1-6, wherein the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising:

[0493] 1 g / L to 50 g / L tryptophan;

[0494] 1 g / L to 100 g / L glucose; manganese; and

[0495] 125 g / L to 200 g / L of one or more nitrogen sources.

[0496] 8. The postbiotic composition of Embodiment 6 or 7, wherein the one or more nitrogen sources comprise yeast extract or peptones.

[0497] 9. The postbiotic composition of any one of Embodiments 1-5, wherein: the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B 12.

[0498] 10. The postbiotic composition of any one of Embodiments 1-5, wherein: PCT / US25 / 16818 21 February 2025 (21.02.2025) the first fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, less than 4 g / L glucose, no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

[0499] 11. The postbiotic composition of any one of Embodiments 1-10, wherein the second fermentate comprises more lAld than the first fermentate.

[0500] 12. The postbiotic composition of any one of Embodiments 1-11, wherein the first fermentate comprises more ILA than the second fermentate.

[0501] 13. The postbiotic composition of any one of Embodiments 1-5, wherein the first fermentate is produced by incubating the bacterial strain under aerobic or microaerobic conditions in a fermentation medium comprising:

[0502] 1 g / L to 12 g / L tryptophan;

[0503] 4 g / L to 15 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12.

[0504] 14. The postbiotic composition of any one of Embodiments 1-5 or 13, wherein the second fermentate is produced by incubating the bacterial strain under the following conditions:

[0505] 1 g / L to 12 g / L tryptophan; less than 4 g / L glucose; and no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

[0506] 15. The postbiotic composition of any one of Embodiments 1-14, wherein a bacterial strain belonging to the species Clostridium sporogenes produces indole- 3 -propionic acid (IPA) in the presence of the postbiotic composition.

[0507] 16. The postbiotic composition of any one of Embodiments 1-15, wherein the first fermentate comprises ILA at a higher amount than lAld. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0508] 17. The postbio tic composition of any one of Embodiments 1-16, wherein the second fermentate comprises lAld at a higher amount than ILA.

[0509] 18. A postbiotic composition comprising two or more fermentates of one or more bacterial strains, wherein a first fermentate comprises indole-3-lactic acid (ILA), and a second fermentate comprises indole-3-carboxaldehyde (lAld).

[0510] 19. The postbiotic composition of any one of Embodiments 1-18, further comprising indole-3-pyruvic acid (IPyA), indole-3-propionic acid (IPA), 3-indoleacrylic acid (IA), and / or indole-3-acetic acid (IAA).

[0511] 20. The postbiotic composition of any one of Embodiments 1-19, wherein the first fermentate and / or the second fermentate further comprises indole- 3 -pyruvic acid (IPyA) and / or indole-3-propionic acid (IPA).

[0512] 21. The postbiotic composition of any one of Embodiments 1-20, wherein the postbiotic composition does not comprise indole-3-acetic acid (IAA)

[0513] 22. The postbiotic composition of any one of Embodiments 1-21, wherein the postbiotic composition further comprises one or more additional fermentates of one or more additional bacterial strains selected from the group consisting of Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans, and Lactiplantibacillus plantarum.

[0514] 23. The postbiotic composition of Embodiment 22, wherein the one or more additional fermentates comprise ILA.

[0515] 24. The postbiotic composition of Embodiment 22 or 23, wherein the one or more additional fermentates comprise lAld.

[0516] 25. The postbiotic composition of any one of Embodiments 22-24, wherein the one or more additional fermentates comprise IPyA and / or IPA. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0517] 26. The postbiotic composition of any one of Embodiments 1-25, wherein the postbiotic composition comprises at least 0.1% (w / w) ILA.

[0518] 27. The postbiotic composition of any one of Embodiments 1-26, wherein the postbiotic composition comprises at least 0.015% (w / w) lAld.

[0519] 28. The postbiotic composition of any one of Embodiments 1-27, wherein the postbiotic composition further comprises IPyA and / or IPA.

[0520] 29. The postbiotic composition of any one of Embodiments 1-28, wherein the postbiotic composition comprises at least 10% w / w IPyA relative to ILA.

[0521] 30. The postbiotic composition of any one of Embodiments 1-29, wherein the postbiotic composition comprises at least 1% w / w IPA relative to lAld.

[0522] 31. The postbiotic composition of any one of Embodiments 1-30, wherein the postbiotic composition comprises at least 10% w / w IAA relative to ILA.

[0523] 32. The postbiotic composition of any one of Embodiments 1-31, wherein the postbiotic composition comprises less than 200% w / w tryptophan relative to ILA.

[0524] 33. The postbiotic composition of any one of Embodiments 1-32, wherein the postbiotic composition does not comprise indoxyl sulfate.

[0525] 34. The postbiotic composition of any one of Embodiments 1-33, wherein the postbiotic composition comprises less than 10% w / w IAA relative to ILA.

[0526] 35. The postbiotic composition of any one of Embodiments 1-34, wherein the postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA.

[0527] 36. The postbiotic composition of any one of Embodiments 1-35, wherein the postbiotic composition comprises less than 0.5% w / w indole-3-acetamide relative to ILA. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0528] 37. The postbiotic composition of any one of Embodiments 1-36, wherein each of the fermentates are in powdered form.

[0529] 38. The postbiotic composition of any one of Embodiments 1-37, wherein each of the fermentates are lyophilized or spray-dried.

[0530] 39. The postbiotic composition of any one of Embodiments 1-38, wherein each of the fermentates are present in an amount of 0.1 to 1000 mg.

[0531] 40. The postbiotic composition of any one of Embodiments 1-39, wherein the postbiotic composition comprises a total amount of fermentates of 0.2 to 2000 mg.

[0532] 41. The postbiotic composition of any one of Embodiments 1-40, wherein the postbiotic composition further comprises an excipient.

[0533] 42. The postbiotic composition of Embodiment 41, wherein the excipient is a tapiocabased maltodextrin.

[0534] 43. The postbiotic composition of any one of Embodiments 1-42, wherein the postbiotic composition is in a form for topical administration.

[0535] 44. The postbiotic composition of any one of Embodiments 1-42, wherein the postbiotic composition is in a form for oral administration.

[0536] 45. The postbiotic composition of any one of Embodiments 1-42, wherein the postbiotic composition is in the form of a dry flowable powder.

[0537] 46. The postbiotic composition of any one of Embodiments 1-45, wherein the postbiotic composition reduces inflammation in a subject.

[0538] 47. The postbiotic composition of any one of Embodiments 1-46, wherein the postbiotic composition treats or prevents a symptom of inflammation in a subject. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0539] 48. The postbiotic composition of any one of Embodiments 1-47, wherein the postbiotic composition treats or prevents diarrhea in a subject.

[0540] 49. The postbiotic composition of any one of Embodiments 1-48, wherein the postbiotic composition treats or prevents vomiting in a subject.

[0541] 50. The postbiotic composition of any one of Embodiments 1-49, wherein the postbiotic composition treats or prevents itching in a subject.

[0542] 51. The postbiotic composition of any one of Embodiments 1-50, wherein the postbiotic composition treats or prevents a symptom of allergy in a subject.

[0543] 52. The postbiotic composition of any one of Embodiments 46-51, wherein the subject is a carnivore.

[0544] 53. The postbiotic composition of any one of Embodiments 46-52, wherein the subject is a mammal.

[0545] 54. The postbiotic composition of any one of Embodiments 46-53, wherein the subject is a domesticated animal.

[0546] 55. The postbiotic composition of any one of Embodiments 46-54, wherein the subject is a dog, cat, rabbit, guinea pig, hamster, or ferret.

[0547] 56. The postbiotic composition of any one of Embodiments 46-55, wherein the subject is a dog.

[0548] 57. A topical preparation comprising the postbiotic composition of any one of Embodiments 1-56.

[0549] 58. The topical preparation of Embodiment 57, wherein the topical preparations is a shampoo (liquid cleansing preparations), a cream (semi-solid, water-based emulsions), an ointment (thicker, greasier preparations usually with an oil base), a spray (liquid preparations in aerosol or pump form), a spot-on solution (liquid preparations specifically designed for PCT / US25 / 16818 21 February 2025 (21.02.2025) application to a single area), a lotion (thinner liquid preparations compared to creams), a gel (semi-solid, typically clear preparations), a foam (aerated preparations that come out as mousse), a balm (waxy, protective preparations), or a powder (dry preparations for dusting).

[0550] 59. The topical preparation of Embodiment 57 or 58, wherein the bacterial fermentate mixture is present in the topical preparation at an incorporation rate of 0.01% to 10%.

[0551] 60. A nutritional supplement or food product comprising the postbiotic composition of any one of Embodiments 1-56.

[0552] 61. The nutritional supplement or food product of Embodiment 60, wherein the nutritional supplement or food product comprises a meat or animal-derived material.

[0553] 62. The nutritional supplement of Embodiment 60 or 61, wherein the meat or animal- derived material is beef, chicken, eggs, turkey, lamb, fish, blood marrow, and / or bone marrow.

[0554] 63. The nutritional supplement or food product of any one of Embodiments 60-62, wherein the nutritional supplement or food product comprises a grain.

[0555] 64. The nutritional supplement or food product of Embodiment 63, wherein the grain is wheat, com, rice, oats, and / or barley.

[0556] 65. The nutritional supplement or food product of any one of Embodiments 60-64, wherein the nutritional supplement or food product comprises a fiber.

[0557] 66. The nutritional supplement or food product of Embodiment 65, wherein the fiber is sugar beet pulp, chicory pulp, chicory, coconut endosperm fiber, wheat fiber, fructooligosaccharides, galactooligosaccharides, and / or inulin.

[0558] 67. The nutritional supplement or food product of any one of Embodiments 60-66, wherein the nutritional supplement or food product comprises an algae. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0559] 68. The nutritional supplement or food product of Embodiment 67, wherein the algae is Ascophyllum nodosum, Spirulina, and / or Fucus vesiculosus.

[0560] 69. The nutritional supplement or food product of any one of Embodiments 60-68, wherein the nutritional supplement or food product comprises a gelatinized starch matrix.

[0561] 70. A method comprising administering the postbiotic composition of any one of Embodiments 1-56, or the nutritional supplement or food product of any one of Embodiments 60-69, to a subject in need thereof.

[0562] 71. The method of Embodiment 70, wherein the administration is oral administration.

[0563] 72. The method of Embodiment 70, wherein the administration is topical administration.

[0564] 73. The method of any one of Embodiments 70-72, wherein the administering:

[0565] (i) reduces inflammation in the subject;

[0566] (ii) treats or prevents a symptom of inflammation in the subject;

[0567] (iii) treats or prevents diarrhea in the subject;

[0568] (iv) treats or prevents vomiting in the subject;

[0569] (v) treats or prevents itching in the subject; and / or

[0570] (vi) treats or prevents a symptom of allergy in the subject.

[0571] 74. The method of any one of Embodiments 70-73, wherein the administering:

[0572] (i) treats or prevents skin inflammation, skin irritation, and / or skin damage in the subject;

[0573] (ii) treats or prevents gut inflammation or gastrointestinal distress in the subject;

[0574] (iii) treats or prevents neuroinflammation in the subject;

[0575] (iv) treats or prevents weight-related inflammation in the subject;

[0576] (v) limits weight gain in the subject;

[0577] (vi) promotes weight loss in the subject;

[0578] (vii) treats or prevents overweight status or obesity in the subject;

[0579] (viii) treats or prevents anxiety in the subject;

[0580] (ix) treats or prevents seasonal allergy in the subject;

[0581] (x) treats or prevents food allergy in the subject; and / or PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0582] (xi) improves coat quality in the subject.

[0583] 75. The method of any one of Embodiments 70-74, wherein the subject is a carnivore.

[0584] 76. The method of any one of Embodiments 70-75, wherein the subject is a mammal.

[0585] 77. The method of any one of Embodiments 70-76, wherein the subject is a domesticated animal.

[0586] 78. The method of any one of Embodiments 70-77, wherein the subject is a dog, cat, rabbit, guinea pig, hamster, or ferret.

[0587] 79. The method of any one of Embodiments 70-78, wherein the subject is a dog.

[0588] 80. The method of any one of Embodiments 70-78, wherein the subject is a cat.

[0589] 81. A method for producing a postbiotic composition, the method comprising:

[0590] (i) fermenting a first bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 (Pediococcus acidlaclici) or SEQ ID NO: 7 (Lactiplantibacillus plantarum) in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA;

[0591] (ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld; and

[0592] (iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld.

[0593] 82. The method of Embodiment 81, wherein the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1.

[0594] 83. The method of Embodiment 81 or 82, wherein the bacterial strain belongs to the species Pediococcus acidilactici. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0595] 84. The method of any one of Embodiments 81-83, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8.

[0596] 85. The method of any one of Embodiments 81-84, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 50.0% to the reference genome of SEQ ID NO: 8.

[0597] 86. The method of any one of Embodiments 81-85, wherein the first fermentate comprises ILA at a higher amount than lAld.

[0598] 87. The method of any one of Embodiments 81-86, wherein the second fermentate comprises lAld at a higher amount than ILA.

[0599] 88. The method of any one of Embodiments 81-87, wherein the first and / or second fermentate further comprises indole-3-pyruvic acid (IPyA) and / or indole-3 -propionic acid (IPA).

[0600] 89. The method of any one of Embodiments 81-88, wherein the first and / or second fermentate further comprises 3 -indoleacrylic acid (IA), and / or indole-3-acetic acid (IAA).

[0601] 90. The method of any one of Embodiments 81-89, further comprising fermenting one or more additional bacterial strains to produce one or more additional fermentates, and combining the one or more additional fermentates with the first and second fermentates or the postbiotic composition, wherein the one or more additional bacterial strains are selected from the group consisting of Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans.

[0602] 91. The method of Embodiment 90, wherein the one or more additional fermentates comprise ILA and / or lAld.

[0603] 92. The method of any one of Embodiments 90-91, wherein the one or more additional fermentates comprise IPyA and / or IPA. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0604] 93. The method of any one of Embodiments 81-92, wherein the postbiotic composition comprises at least 0.1% (w / w) ILA.

[0605] 94. The method of any one of Embodiments 81-93, wherein the postbiotic composition comprises at least 0.015% (w / w) lAld.

[0606] 95. The method of any one of Embodiments 81-94, wherein the postbiotic composition further comprises IPyA and / or IPA.

[0607] 96. The method of any one of Embodiments 81-95, wherein the postbiotic composition comprises at least 10% w / w IPyA relative to ILA.

[0608] 97. The method of any one of Embodiments 81-96, wherein the postbiotic composition comprises at least 1% w / w IPA relative to lAld.

[0609] 98. The method of any one of Embodiments 81-97, wherein the postbiotic composition comprises less than 200% w / w tryptophan relative to ILA.

[0610] 99. The method of any one of Embodiments 81-98, wherein the postbiotic composition does not comprise indole.

[0611] 100. The method of any one of Embodiments 81-99, wherein the postbiotic composition does not comprise indoxyl sulfate.

[0612] 101. The method of any one of Embodiments 81-100, wherein the postbiotic composition comprises less than 10% w / w IAA relative to ILA.

[0613] 102. The method of any one of Embodiments 81-101, wherein the postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA.

[0614] 103. The method of any one of Embodiments 81-102, wherein the postbiotic composition comprises less than 0.5% w / w indole-3-acetamide relative to ILA. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0615] 104. The method of any one of Embodiments 81-103, wherein the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising:

[0616] 1 g / L to 50 g / L tryptophan;

[0617] 100 g / L to 200 g / L glucose; manganese; and

[0618] 10 g / L to 125 g / L of one or more nitrogen sources.

[0619] 105. The method of any one of Embodiments 81-104, wherein the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising:

[0620] 1 g / L to 50 g / L tryptophan;

[0621] 1 g / L to 100 g / L glucose; manganese; and

[0622] 125 g / L to 200 g / L of one or more nitrogen sources.

[0623] 106. The method of Embodiment 104 or 105, wherein the one or more nitrogen sources comprise yeast extract or peptones.

[0624] 107. The method of any one of Embodiments 81-103, wherein: the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B 12.

[0625] 108. The method of any one of Embodiments 81-103, wherein: the first fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, less than 4 g / L PCT / US25 / 16818 21 February 2025 (21.02.2025) glucose, no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

[0626] 109. The method of any one of Embodiments 81-108, wherein the second fermentate comprises more lAld than the first fermentate.

[0627] 110. The method of any one of Embodiments 81-109, wherein the first fermentate comprises more ILA than the second fermentate.

[0628] 111. The method of any one of Embodiments 81-103, wherein the first fermentation medium and / or the second fermentation medium comprises tryptophan at a concentration of 1 to 12 g / L.

[0629] 112. The method of any one of Embodiments 81-103 or 111, wherein the first fermentation medium comprises glucose at a concentration of at least 4 g / L.

[0630] 113. The method of any one of Embodiments 81-103 or 111-112, wherein the first fermentation medium comprises glucose at a concentration of at least 4 to 15 g / L.

[0631] 114. The method of any one of Embodiments 81-103 or 111-113, wherein the second fermentation medium comprises glucose at a concentration of less than 4 g / L.

[0632] 115. The method of any one of Embodiments 81-103 or 111-114, wherein the first fermentation medium comprises polysorbate 80, manganese, and / or vitamin B12.

[0633] 116. The method of any one of Embodiments 81-103 or 111-115, wherein the second fermentation medium does not comprise polysorbate 80, does not comprise manganese, and does not comprise vitamin B12.

[0634] 117. The method of any one of Embodiments 81-116, wherein the fermenting of (i) and / or the fermenting of (ii) is for 2 to 7 days.

[0635] 118. The method of any one of Embodiments 81-117, wherein the fermenting of (i) and / or the fermenting of (ii) is under aerobic or microaerobic conditions. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0636] 119. The method of any one of Embodiments 81-118, further comprising separating bacterial cells from the first fermentate and / or second fermentate before combining the first and second fermentates.

[0637] 120. The method of any one of Embodiments 81-119, further comprising heat killing bacterial strains in the first fermentate and / or second fermentate before combining the first and second fermentates.

[0638] 121. The method of any one of Embodiments 81-120, further comprising concentrating the first fermentate and / or second fermentate before combining the first and second fermentates.

[0639] 122. The method of any one of Embodiments 81-121, further comprising drying the first fermentate and / or second fermentate before combining the first and second fermentates.

[0640] 123. A postbio tic composition produced by the method of any one of Embodiments 81- 122.

[0641] 201. A composition comprising a bacterial fermentate mixture, the bacterial fermentate mixture comprising a first fermentate and a second fermentate of a bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1, wherein the first fermentate comprises more indole- 3 -lactic acid (ILA) than indole-3- carboxaldehyde (lAld) and the second fermentate comprises more lAld than the first fermentate, wherein each of the fermentates is in powder form, wherein the composition is a nutritional supplement or food product.

[0642] 202. The composition of Embodiment 201, wherein:

[0643] (i) the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan; and

[0644] (ii) the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a second fermentation medium comprising 1 g / L to 50 g / L tryptophan. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0645] 203. The composition of Embodiment 202, wherein:

[0646] (i) the first fermentation medium comprises 100 g / L to 200 g / L glucose and 10 g / L to 125 g / L of one or more nitrogen sources; and

[0647] (ii) the second fermentation medium comprises 1 g / L to 100 g / L glucose and 125 g / L to 200 g / L of one or more nitrogen sources, wherein the first fermentation medium comprises a higher glucose concentration than the second fermentation medium, and the second fermentation medium comprises a higher nitrogen source concentration than the first fermentation medium.

[0648] 204. The composition of Embodiment 203, wherein each of the first fermentation medium and the second fermentation medium comprises manganese.

[0649] 205. The composition of Embodiment 201, wherein the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to the nucleotide sequence of SEQ ID NO: 1.

[0650] 206. The composition of Embodiment 205, wherein the bacterial strain belongs to the species Pediococcus acidilactici.

[0651] 207. The composition of Embodiment 206, wherein the bacterial fermentate mixture consists of the first fermentate and the second fermentate.

[0652] 208. The composition of Embodiment 201, wherein the nutritional supplement or food product comprises maltodextrin.

[0653] 209. The composition of Embodiment 201, wherein each of the fermentates is lyophilized or spray-dried.

[0654] 210. The composition of Embodiment 201, wherein the nutritional supplement or food product is in the form of gelatinized starch matrix, extruded or injection-molded treat, extruded or injection-molded chew, powder meal topper, water additive, dry kibble or food, wet food, frozen food, liquid spray, peanut butter spread, or soft jerky chew. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0655] 211. The composition of Embodiment 201, further comprising indole-3-pyruvic acid (IPyA), indole- 3 -propionic acid (IPA), 3-indoleacrylic acid (IA), and / or indole-3-acetic acid (IAA).

[0656] 212. The composition of Embodiment 201, wherein the nutritional supplement or food product is formulated for administration to a canine subject at an ILA dose of 0.8 pg / kg per day to 8 mg / kg per day and an lAld dose of 0.08 pg / kg per day to 800 pg / kg per day.

[0657] 213. The composition of Embodiment 201, wherein each of the first and second fermentates is present in the nutritional supplement or food product at an amount of 0.1 to 1000 mg, such that the bacterial fermentate mixture is present in the nutritional supplement or food product in an amount of 0.2 to 2000 mg.

[0658] 214. The composition of Embodiment 201, wherein the composition treats or prevents a symptom of inflammation in a subject.

[0659] 215. The composition of Embodiment 214, wherein the composition treats or prevents one or more symptoms of atopic dermatitis in the subject.

[0660] 216. The composition of Embodiment 214, wherein the composition treats or prevents one or more symptoms of allergy in the subject.

[0661] 217. The composition of Embodiment 214, wherein the composition treats or prevents itching in the subject.

[0662] 218. The composition of Embodiment 214, wherein the composition increases diversity of the intestinal microbiome in a subject as measured by Shannon diversity index of zero-radius operational taxonomic units (OTUs) present in a fecal sample of the subject.

[0663] 219. The composition of Embodiment 214, wherein the subject is a dog or cat.

[0664] 220. A composition comprising a bacterial fermentate mixture, the bacterial fermentate mixture comprising a first fermentate and a second fermentate of a bacterial strain belonging PCT / US25 / 16818 21 February 2025 (21.02.2025) to the species Pediococcus acidilactici and comprising a 16S rDNA sequence with at least 99% sequence identity to the nucleotide sequence of SEQ ID NO: 1, wherein the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan, 100 g / L to 200 g / L glucose, 10 g / L to 125 g / L of one or more nitrogen sources, and manganese, such that the first fermentate comprises more indole-3-lactic acid (ILA) than indole-3-carboxaldehyde (lAld), wherein the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a second fermentation medium comprising 1 g / L to 50 g / L tryptophan, 1 g / L to 100 g / L glucose, 125 g / L to 200 g / L of one or more nitrogen sources, and manganese, such that the second fermentate comprises more lAld than the first fermentate, wherein the first fermentation medium comprises a higher glucose concentration than the second fermentation medium, and the second fermentation medium comprises a higher nitrogen source concentration than the first fermentation medium, wherein each of the fermentates is in powder form, wherein the composition is a nutritional supplement or food product.

[0665] 301. A method of treating or preventing one or more symptoms of inflammation in a subject in need thereof, the method comprising orally administering to the subject a nutritional supplement or food product comprising a bacterial fermentate mixture, the bacterial fermentate mixture comprising a first fermentate and a second fermentate of a bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1, wherein the first fermentate comprises more indole- 3 -lactic acid (ILA) than indole-3- carboxaldehyde (lAld) and the second fermentate comprises more lAld than the first fermentate.

[0666] 302. The method of Embodiment 301, wherein:

[0667] (i) the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan; and

[0668] (ii) the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a second fermentation medium comprising 1 g / L to 50 g / L tryptophan. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0669] 303. The method of Embodiment 302, wherein:

[0670] (i) the first fermentation medium comprises 100 g / L to 200 g / L glucose and 10 g / L to 125 g / L of one or more nitrogen sources; and

[0671] (ii) the second fermentation medium comprises 1 g / L to 100 g / L glucose and 125 g / L to 200 g / L of one or more nitrogen sources, wherein the first fermentation medium comprises a higher glucose concentration than the second fermentation medium, and the second fermentation medium comprises a higher nitrogen source concentration than the first fermentation medium.

[0672] 304. The method of Embodiment 303, wherein each of the first fermentation medium and the second fermentation medium comprises manganese.

[0673] 305. The method of Embodiment 301, wherein the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to the nucleotide sequence of SEQ ID NO: 1.

[0674] 306. The method of Embodiment 305, wherein the bacterial strain belongs to the species Pediococcus acidilactici.

[0675] 307. The method of Embodiment 306, wherein the bacterial fermentate mixture consists of the first fermentate and the second fermentate.

[0676] 308. The method of Embodiment 301, wherein the subject is a dog or cat.

[0677] 309. The method of Embodiment 301, wherein the subject is a dog.

[0678] 310. The method of Embodiment 309, wherein the bacterial fermentate mixture stimulates a canine aryl hydrocarbon receptor (AhR).

[0679] 311. The method of Embodiment 309, wherein the bacterial fermentate mixture activates a nuclear factor erythroid 2-related factor 2 (Nrf2).

[0680] 312. The method of Embodiment 309, wherein the bacterial fermentate mixture inhibits production of interferon-gamma (IFN-y) and / or tumor necrosis factor alpha (TNF-a) in the dog. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0681] 313. The method of Embodiment 309, wherein the administering reduces itching severity in the dog.

[0682] 314. The method of Embodiment 309, wherein the administering improves coat quality in the dog.

[0683] 315. The method of Embodiment 309, wherein the administering increases, in an intestinal microenvironment of the dog, abundance of one or more microorganisms that produce indole-3-proprionic acid and / or butyrate.

[0684] 316. The method of Embodiment 301, wherein each of the first and second fermentates is present in the nutritional supplement or food product at an amount of 0.1 to 1000 mg.

[0685] 317. The method of Embodiment 301, wherein the bacterial fermentate mixture is present in the nutritional supplement or food product at an incorporation rate of 0.01% to 10% w / w.

[0686] 318. The method of Embodiment 301, wherein the nutritional supplement or food product is in the form of a gelatinized starch matrix, extruded or injection-molded treat, extruded or injection-molded chew, powder meal topper, water additive, dry kibble or food, wet food, frozen food, liquid spray, peanut butter spread, or soft jerky chew.

[0687] 319. A method of reducing itching severity in a canine subject in need thereof, the method comprising orally administering to the subject a nutritional supplement or food product comprising a bacterial fermentate mixture, the bacterial fermentate mixture comprising a first fermentate and a second fermentate of a bacterial strain belonging to the species Pediococcus acidilactici and comprising a 16S rDNA sequence with at least 99% sequence identity to the nucleotide sequence of SEQ ID NO: 1, wherein the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a first fermentation medium comprising 1 g / L to 50 g / L tryptophan, 100 g / L to 200 g / L glucose, 10 g / L to 125 g / L of one or more nitrogen sources, and manganese, such that the first fermentate comprises more indole-3-lactic acid (ILA) than indole-3-carboxaldehyde (lAld), PCT / US25 / 16818 21 February 2025 (21.02.2025) wherein the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a second fermentation medium comprising 1 g / L to 50 g / L tryptophan, 1 g / L to 100 g / L glucose, 125 g / L to 200 g / L of one or more nitrogen sources, and manganese, such that the second fermentate comprises more lAld than the first fermentate, wherein the first fermentation medium comprises a higher glucose concentration than the second fermentation medium, and the second fermentation medium comprises a higher nitrogen source concentration than the first fermentation medium.

[0688] 320. The method of Embodiment 319, wherein the bacterial fermentate mixture reduces scratching frequency in the canine subject as measured by pruritus visual analog scale (PVAS).

[0689] EQUIVALENTSAND SCOPE

[0690] While several inventive embodiments have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and / or structures for performing the function and / or obtaining the results and / or one or more of the advantages described herein, and each of such variations and / or modifications is deemed to be within the scope of the inventive embodiments described herein. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and / or configurations will depend upon the specific application or applications for which the inventive teachings is / are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific inventive embodiments described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, inventive embodiments may be practiced otherwise than as specifically described and claimed. Inventive embodiments of the present disclosure are directed to each individual feature, system, article, material, kit, and / or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and / or methods, if such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent, is included within the inventive scope of the present disclosure. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0691] All definitions, as defined and used herein, should be understood to control over dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0692] All references, patents and patent applications disclosed herein are incorporated by reference with respect to the subject matter for which each is cited, which in some cases may encompass the entirety of the document.

[0693] The indefinite articles “a” and “an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean “at least one.”

[0694] The phrase “and / or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with “and / or” should be construed in the same fashion, i.e., “one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the “and / or” clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to “A and / or B”, when used in conjunction with open-ended language such as “comprising” can refer, in some embodiments, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc. As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of’ or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e. “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law.

[0695] As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also PCT / US25 / 16818 21 February 2025 (21.02.2025) allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and / or B”) can refer, in some embodiments, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc. Each possibility represents a separate embodiment of the present invention.

[0696] It should be understood that, unless clearly indicated to the contrary, the disclosure of numerical values and ranges of numerical values in the specification includes both i) the exact value(s) or range specified, and ii) values that are “about” the value(s) or ranges specified (e.g., values or ranges falling within a reasonable range (e.g., about 10% similar)) as would be understood by a person of ordinary skill in the art.

[0697] It should also be understood that, unless clearly indicated to the contrary, in any methods disclosed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are disclosed.

[0698] In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of’ and “consisting essentially of’ shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

[0699] EXAMPLES

[0700] Examples 1-7, 21, and 22 describe experiments that have actually been conducted. Examples 8-20 are prophetic examples. Where examples refer to the postbiotic composition of Example 2, containing the first and second fermentates of Strain 1, by the term “Strain 1 Postbiotic Fermentate Mixture”, mixtures of fermentates of other bacteria (e.g., Strains 2-7) may be applied to the described conditions in similar manners. Where examples refer to the postbiotic composition of Example 2, containing the first and second fermentates of Strain 1, PCT / US25 / 16818 21 February 2025 (21.02.2025) by the term “Canine Immune Health Postbiotic” or “(CIHP)”, the skilled artisan will appreciate that the utility of such compositions is not limited to canines, as stimulation of the aryl hydrocarbon receptor and / or limiting inflammation by use of the fermentate mixture is beneficial in the context of other species, such as other domesticated animals (e.g., cats) and humans.

[0701] Example 1: Production of indole-3-lactic acid (ILA) and indole-3-carboxaldehyde (lAld) by bacterial fermentation.

[0702] Bacterial strain Pediococcus acidilactici (having a 16S rDNA sequence of SEQ ID NO: 1) was fermented in tryptophan-containing medium at varying concentrations of glucose, and titers of ILA and lAld were measured in bacterial fermentates. It was discovered that while P. acidilactici produced both ILA and lAld, the amounts of each compound produced varied with glucose concentration, with the greatest ILA production occurring at glucose concentrations above 4 g / L, and the most lAld production occurring at glucose concentrations below 4 g / L (FIG. 1). Eermentation of P. acidilactici at exemplary glucose concentrations yielded fermentates containing 600 mg / L ILA (FIG. 2A) or 15.0 mg / L (FIG. 2B).

[0703] Next, P. acidilactici was fermented in separate reactions having the glucose concentrations determined optimal for ILA or lAld production, with or without supplementation of the media with a combination of (i) polysorbate 80, (ii) manganese, and (iii) vitamin B12. While this supplementation increased production of ILA (FIG. 3A), it decreased production of lAld (FIG. 3B). The effect of oxygen exposure was also investigated, revealing that microaerobic conditions improved yields of both ILA and lAld, while both compounds were produced less under anaerobic conditions (FIGs. 4A and 4B).

[0704] The utility of ILA and lAld in multiple host was validated by an in vitro assay. Human or canine cell lines expressing AhR were exposed to ILA or lAld at 200 pM, and AhR activation was assayed using in vitro luminescence. AhR activation was observed using both compounds (FIG. 5), indicating the benefit of both ILA and lAld for AhR agonism.

[0705] Example 2: Improved activation of canine AhR by a mixture of fermentates of Strain 1.

[0706] The ability of a postbiotic to activate canine AhR was validated by an in vitro assay. The postbiotic composition comprised a first fermentate and a second fermentate of Strain 1, belonging to Pediococcus acidilactici, whose 16S rRNA gene sequence is provided by SEQ ID NO: 1, and genome is provided by SEQ ID NO: 8. The first fermentate contained ILA, and the second fermentate contained lAld. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0707] AhR activation was assessed using a luciferase reporter assay; a luminescent signal is generated upon activation of AhR. The negative control was water and the positive control was 2 mg / mL methylcholanthrene, an AhR agonist. The AhR reporter canine cell lines were treated for 24 hours with controls, the postbiotic, or a blend of pure ILA and lAld at the same ratio and concentrations as the postbiotic. Cell viability was measured, cells were lysed, and luciferase activity was measured using a luminometer upon addition of a luminescence detection reagent (luciferin).

[0708] The luminescence light intensity (corrected by number of viable cells) is directly proportional to the extent of AhR activation. The postbiotic unexpectedly caused a fold activation of canine AhR that was lOx higher than a blend of pure ILA and lAld (FIG. 6).

[0709] Example 3: Improved reduction of pro-inflammatory cytokines by a mixture of fermentates of Strain 1.

[0710] The ability of the postbiotic of Example 2, containing the first and second fermentates of Strain 1, to modulate inflammation, in particular by reducing pro-inflammatory cytokines, was validated by an in vitro lymphocyte stimulation assay. Peripheral blood mononuclear cells (PBMCS) were harvested from one human donor and treated with controls, the postbiotic, or a blend of pure ILA and lAld at the same ratio and concentrations as the postbiotic. The positive control was the reference inhibitor dexamethasone at 100 nM. The postbiotic and pure ILA / IAld blend were tested at a range of concentrations. PBMCs were seeded at a density of 2xl05cells / well in culture medium and incubated for 1 hour prior to the addition of test agents. Test agents and controls were added in triplicate and incubated for 1 hour. Next, the inducing agent phytohemagglutinin-L was added at 10 pg / mL, followed by incubation for 48 hours. Supernatants were then collected for analysis, and cytokine levels were determined using Luminex® methodology.

[0711] Levels of each cytokine were interpolated from standard curves using non-linear regression analysis and normalized to the vehicle control. The postbiotic significantly inhibited the release of the pro-inflammatory cytokines interferon-gamma (IFN-y) and tumor necrosis factor alpha (TNF-a) at all concentrations tested. The postbiotic unexpectedly outperformed the mixture of pure ILA / IAld blend with respect to both cytokines (FIGs. 7A and 7B), with only minimal detection of IFN-y (FIG. 7A), and inhibition of TNF-a production where the mixture of pure IFA+IAld failed to inhibit TNF-a production. PCT / US25 / 16818 21 February 2025 (21.02.2025)

[0712] Example 4: Reduction of scratching behavior in dogs following administration of a mixture of fermentates of Strain 1.

[0713] A double-blind, multi-group controlled study was conducted to evaluate the efficacy of the postbiotic of Example 2, containing the first and second fermentates of Strain 1 in reducing canine itching. Thirty dogs, evenly divided into postbiotic and placebo groups, participated over 50 days, including a 14-day baseline observation and a 28-day treatment phase. The postbiotic or placebo was administered twice daily as a meal topper, with stratified randomization ensuring balanced groups by sex and baseline scratching levels. The primary biomarker was scratching time, measured in seconds per day using a wearable activity monitor, with the rolling weekly average used to assess efficacy. Secondary biomarkers included licking time (also tracked by the wearable activity monitor), skin metagenomic analysis from samples taken on Days 0 and 28 to track microbiome shifts, and fecal metagenomic analysis from the same days to track microbiome shifts. Skin samples were taken from three locations (armpit, groin, and paw), while fecal samples were collected from first-evacuated feces of the day, refrigerated, and shipped on ice packs. Human-reported metrics, collected biweekly, included perceptions of coat quality (using a standard 4-point scale, with lowest score indicating poor quality coat), and i...

Claims

PCT / US25 / 16818 21 February 2025 (21.02.2025)CLAIMSWhat is claimed is:

1. A postbiotic composition comprising a first fermentate and a second fermentate of a bacterial strain, the bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 (Pediococcus acidilaclici) or SEQ ID NO: 7 (Lactiplantibacillus plantarum), and wherein the first fermentate comprises indole-3 -lactic acid (ILA) and the second fermentate comprises indole-3-carboxaldehyde (lAld).

2. The postbiotic composition of claim 1, wherein the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1.

3. The postbiotic composition of claim 1 or 2, wherein the bacterial strain belongs to the species Pediococcus acidilactici.

4. The postbiotic composition of any one of claims 1-3, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8.

5. The postbiotic composition of any one of claims 1-4, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 50.0% to the reference genome of SEQ ID NO: 8.

6. The postbiotic composition of any one of claims 1-5, wherein the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising:1 g / L to 50 g / L tryptophan;100 g / L to 200 g / L glucose; manganese; and10 g / L to 125 g / L of one or more nitrogen sources.PCT / US25 / 16818 21 February 2025 (21.02.2025)7. The postbiotic composition of any one of claims 1-6, wherein the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising:1 g / L to 50 g / L tryptophan;1 g / L to 100 g / L glucose; manganese; and125 g / L to 200 g / L of one or more nitrogen sources.

8. The postbiotic composition of claim 6 or 7, wherein the one or more nitrogen sources comprise yeast extract or peptones.

9. The postbiotic composition of any one of claims 1-5, wherein: the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B 12.

10. The postbiotic composition of any one of claims 1-5, wherein: the first fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, less than 4 g / L glucose, no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

11. The postbiotic composition of any one of claims 1-10, wherein the second fermentate comprises more lAld than the first fermentate.

12. The postbiotic composition of any one of claims 1-11, wherein the first fermentate comprises more ILA than the second fermentate.PCT / US25 / 16818 21 February 2025 (21.02.2025)13. The postbiotic composition of any one of claims 1-5, wherein the first fermentate is produced by incubating the bacterial strain under aerobic or microaerobic conditions in a fermentation medium comprising:1 g / L to 12 g / L tryptophan;4 g / L to 15 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B12.

14. The postbiotic composition of any one of claims 1-5 or 13, wherein the second fermentate is produced by incubating the bacterial strain under the following conditions:1 g / L to 12 g / L tryptophan; less than 4 g / L glucose; and no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

15. The postbiotic composition of any one of claims 1-14, wherein a bacterial strain belonging to the species Clostridium sporogenes produces indole-3 -propionic acid (IPA) in the presence of the postbiotic composition.

16. The postbiotic composition of any one of claims 1-15, wherein the first fermentate comprises ILA at a higher amount than lAld.

17. The postbiotic composition of any one of claims 1-16, wherein the second fermentate comprises lAld at a higher amount than ILA.

18. A postbiotic composition comprising two or more fermentates of one or more bacterial strains, wherein a first fermentate comprises indole-3-lactic acid (ILA), and a second fermentate comprises indole-3-carboxaldehyde (lAld).

19. The postbiotic composition of any one of claims 1-18, further comprising indole-3- pyruvic acid (IPyA), indole-3-propionic acid (IPA), 3-indoleacrylic acid (IA), and / or indole- 3-acetic acid (IAA).PCT / US25 / 16818 21 February 2025 (21.02.2025)20. The postbiotic composition of any one of claims 1-19, wherein the first fermentate and / or the second fermentate further comprises indole- 3 -pyruvic acid (IPyA) and / or indole-3- propionic acid (IPA).

21. The postbiotic composition of any one of claims 1-20, wherein the postbiotic composition does not comprise indole-3-acetic acid (IAA)22. The postbiotic composition of any one of claims 1-21, wherein the postbiotic composition further comprises one or more additional fermentates of one or more additional bacterial strains selected from the group consisting of Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans, and Lactiplantibacillus plantarum.

23. The postbiotic composition of claim 22, wherein the one or more additional fermentates comprise ILA.

24. The postbiotic composition of claim 22 or 23, wherein the one or more additional fermentates comprise lAld.

25. The postbiotic composition of any one of claims 22-24, wherein the one or more additional fermentates comprise IPyA and / or IPA.

26. The postbiotic composition of any one of claims 1-25, wherein the postbiotic composition comprises at least 0.1% (w / w) ILA.

27. The postbiotic composition of any one of claims 1-26, wherein the postbiotic composition comprises at least 0.015% (w / w) lAld.

28. The postbiotic composition of any one of claims 1-27, wherein the postbiotic composition further comprises IPyA and / or IPA.

29. The postbiotic composition of any one of claims 1-28, wherein the postbiotic composition comprises at least 10% w / w IPyA relative to ILA.PCT / US25 / 16818 21 February 2025 (21.02.2025)30. The postbiotic composition of any one of claims 1-29, wherein the postbiotic composition comprises at least 1% w / w IPA relative to lAld.

31. The postbiotic composition of any one of claims 1-30, wherein the postbiotic composition comprises at least 10% w / w IAA relative to ILA.

32. The postbiotic composition of any one of claims 1-31, wherein the postbiotic composition comprises less than 200% w / w tryptophan relative to ILA.

33. The postbiotic composition of any one of claims 1-32, wherein the postbiotic composition does not comprise indoxyl sulfate.

34. The postbiotic composition of any one of claims 1-33, wherein the postbiotic composition comprises less than 10% w / w IAA relative to ILA.

35. The postbiotic composition of any one of claims 1-34, wherein the postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA.

36. The postbiotic composition of any one of claims 1-35, wherein the postbiotic composition comprises less than 0.5% w / w indole-3-acetamide relative to ILA.

37. The postbiotic composition of any one of claims 1-36, wherein each of the fermentates are in powdered form.

38. The postbiotic composition of any one of claims 1-37, wherein each of the fermentates are lyophilized or spray-dried.

39. The postbiotic composition of any one of claims 1-38, wherein each of the fermentates are present in an amount of 0.1 to 1000 mg.

40. The postbiotic composition of any one of claims 1-39, wherein the postbiotic composition comprises a total amount of fermentates of 0.2 to 2000 mg.PCT / US25 / 16818 21 February 2025 (21.02.2025)41. The postbiotic composition of any one of claims 1-40, wherein the postbiotic composition further comprises an excipient.

42. The postbiotic composition of claim 41, wherein the excipient is a tapioca-based maltodextrin.

43. The postbiotic composition of any one of claims 1-42, wherein the postbiotic composition is in a form for topical administration.

44. The postbiotic composition of any one of claims 1-42, wherein the postbiotic composition is in a form for oral administration.

45. The postbiotic composition of any one of claims 1-42, wherein the postbiotic composition is in the form of a dry flowable powder.

46. The postbiotic composition of any one of claims 1-45, wherein the postbiotic composition reduces inflammation in a subject.

47. The postbiotic composition of any one of claims 1-46, wherein the postbiotic composition treats or prevents a symptom of inflammation in a subject.

48. The postbiotic composition of any one of claims 1-47, wherein the postbiotic composition treats or prevents diarrhea in a subject.

49. The postbiotic composition of any one of claims 1-48, wherein the postbiotic composition treats or prevents vomiting in a subject.

50. The postbiotic composition of any one of claims 1-49, wherein the postbiotic composition treats or prevents itching in a subject.

51. The postbiotic composition of any one of claims 1-50, wherein the postbiotic composition treats or prevents a symptom of allergy in a subject.PCT / US25 / 16818 21 February 2025 (21.02.2025)52. The postbiotic composition of any one of claims 46-51, wherein the subject is a carnivore.

53. The postbiotic composition of any one of claims 46-52, wherein the subject is a mammal.

54. The postbiotic composition of any one of claims 46-53, wherein the subject is a domesticated animal.

55. The postbiotic composition of any one of claims 46-54, wherein the subject is a dog, cat, rabbit, guinea pig, hamster, or ferret.

56. The postbiotic composition of any one of claims 46-55, wherein the subject is a dog.

57. A topical preparation comprising the postbiotic composition of any one of claims 1- 56.

58. The topical preparation of claim 57, wherein the topical preparations is a shampoo (liquid cleansing preparations), a cream (semi-solid, water-based emulsions), an ointment (thicker, greasier preparations usually with an oil base), a spray (liquid preparations in aerosol or pump form), a spot-on solution (liquid preparations specifically designed for application to a single area), a lotion (thinner liquid preparations compared to creams), a gel (semi-solid, typically clear preparations), a foam (aerated preparations that come out as mousse), a balm (waxy, protective preparations), or a powder (dry preparations for dusting).

59. The topical preparation of claim 57 or 58, wherein the bacterial fermentate mixture is present in the topical preparation at an incorporation rate of 0.01% to 10%.

60. A nutritional supplement or food product comprising the postbiotic composition of any one of claims 1-56.

61. The nutritional supplement or food product of claim 60, wherein the nutritional supplement or food product comprises a meat or animal-derived material.PCT / US25 / 16818 21 February 2025 (21.02.2025)62. The nutritional supplement of claim 60 or 61, wherein the meat or animal-derived material is beef, chicken, eggs, turkey, lamb, fish, blood marrow, and / or bone marrow.

63. The nutritional supplement or food product of any one of claims 60-62, wherein the nutritional supplement or food product comprises a grain.

64. The nutritional supplement or food product of claim 63, wherein the grain is wheat, com, rice, oats, and / or barley.

65. The nutritional supplement or food product of any one of claims 60-64, wherein the nutritional supplement or food product comprises a fiber.

66. The nutritional supplement or food product of claim 65, wherein the fiber is sugar beet pulp, chicory pulp, chicory, coconut endosperm fiber, wheat fiber, fructooligosaccharides, galactooligosaccharides, and / or inulin.

67. The nutritional supplement or food product of any one of claims 60-66, wherein the nutritional supplement or food product comprises an algae.

68. The nutritional supplement or food product of claim 67, wherein the algae is Ascophyllum nodosum, Spirulina, and / or Fucus vesiculosus.

69. The nutritional supplement or food product of any one of claims 60-68, wherein the nutritional supplement or food product comprises a gelatinized starch matrix.

70. A method comprising administering the postbiotic composition of any one of claims 1-56, or the nutritional supplement or food product of any one of claims 60-69, to a subject in need thereof.

71. The method of claim 70, wherein the administration is oral administration.

72. The method of claim 70, wherein the administration is topical administration.

73. The method of any one of claims 70-72, wherein the administering:PCT / US25 / 16818 21 February 2025 (21.02.2025)(i) reduces inflammation in the subject;(ii) treats or prevents a symptom of inflammation in the subject;(iii) treats or prevents diarrhea in the subject;(iv) treats or prevents vomiting in the subject;(v) treats or prevents itching in the subject; and / or(vi) treats or prevents a symptom of allergy in the subject.

74. The method of any one of claims 70-73, wherein the administering:(i) treats or prevents skin inflammation, skin irritation, and / or skin damage in the subject;(ii) treats or prevents gut inflammation or gastrointestinal distress in the subject;(iii) treats or prevents neuroinflammation in the subject;(iv) treats or prevents weight-related inflammation in the subject;(v) limits weight gain in the subject;(vi) promotes weight loss in the subject;(vii) treats or prevents overweight status or obesity in the subject;(viii) treats or prevents anxiety in the subject;(ix) treats or prevents seasonal allergy in the subject;(x) treats or prevents food allergy in the subject; and / or(xi) improves coat quality in the subject.

75. The method of any one of claims 70-74, wherein the subject is a carnivore.

76. The method of any one of claims 70-75, wherein the subject is a mammal.

77. The method of any one of claims 70-76, wherein the subject is a domesticated animal.

78. The method of any one of claims 70-77, wherein the subject is a dog, cat, rabbit, guinea pig, hamster, or ferret.

79. The method of any one of claims 70-78, wherein the subject is a dog.

80. The method of any one of claims 70-78, wherein the subject is a cat.PCT / US25 / 16818 21 February 2025 (21.02.2025)81. A method for producing a postbiotic composition, the method comprising:(i) fermenting a first bacterial strain comprising a 16S rDNA sequence with at least 97% sequence identity to the nucleotide sequence of SEQ ID NO: 1 (Pediococcus acidlaclici) or SEQ ID NO: 7 (Lactiplantibacillus plantarum) in a first fermentation medium under conditions for production of indole-3-lactic acid (ILA), thereby producing a first fermentate comprising ILA;(ii) fermenting the first bacterial strain in a second fermentation medium under conditions for production of indole-3-carboxaldehyde (lAld), thereby producing a second fermentate comprising lAld; and(iii) combining the first and second fermentates to produce a postbiotic composition comprising ILA and lAld.

82. The method of claim 81, wherein the bacterial strain comprises a 16S rDNA sequence with at least 99% sequence identity to SEQ ID NO: 1.

83. The method of claim 81 or 82, wherein the bacterial strain belongs to the species Pediococcus acidilactici.

84. The method of any one of claims 81-83, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 20.0% to the reference genome of SEQ ID NO: 8.

85. The method of any one of claims 81-84, wherein the bacterial strain comprises a genome with at least 95.0% average nucleotide identity (ANI) and an alignment fraction (AF) of at least 50.0% to the reference genome of SEQ ID NO: 8.

86. The method of any one of claims 81-85, wherein the first fermentate comprises ILA at a higher amount than lAld.

87. The method of any one of claims 81-86, wherein the second fermentate comprises I Aid at a higher amount than ILA.

88. The method of any one of claims 81-87, wherein the first and / or second fermentate further comprises indole- 3 -pyruvic acid (IPyA) and / or indole-3-propionic acid (IPA).PCT / US25 / 16818 21 February 2025 (21.02.2025)89. The method of any one of claims 81-88, wherein the first and / or second fermentate further comprises 3-indoleacrylic acid (IA), and / or indole-3-acetic acid (IAA).

90. The method of any one of claims 81-89, further comprising fermenting one or more additional bacterial strains to produce one or more additional fermentates, and combining the one or more additional fermentates with the first and second fermentates or the postbiotic composition, wherein the one or more additional bacterial strains are selected from the group consisting of Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus velezensis, Weizmannia coagulans.

91. The method of claim 90, wherein the one or more additional fermentates comprise ILA and / or lAld.

92. The method of any one of claims 90-91, wherein the one or more additional fermentates comprise IPyA and / or IPA.

93. The method of any one of claims 81-92, wherein the postbiotic composition comprises at least 0.1% (w / w) ILA.

94. The method of any one of claims 81-93, wherein the postbiotic composition comprises at least 0.015% (w / w) lAld.

95. The method of any one of claims 81-94, wherein the postbiotic composition further comprises IPyA and / or IPA.

96. The method of any one of claims 81-95, wherein the postbiotic composition comprises at least 10% w / w IPyA relative to ILA.

97. The method of any one of claims 81-96, wherein the postbiotic composition comprises at least 1% w / w IPA relative to lAld.

98. The method of any one of claims 81-97, wherein the postbiotic composition comprises less than 200% w / w tryptophan relative to ILA.PCT / US25 / 16818 21 February 2025 (21.02.2025)99. The method of any one of claims 81-98, wherein the postbiotic composition does not comprise indole.

100. The method of any one of claims 81-99, wherein the postbiotic composition does not comprise indoxyl sulfate.

101. The method of any one of claims 81-100, wherein the postbiotic composition comprises less than 10% w / w IAA relative to ILA.

102. The method of any one of claims 81-101, wherein the postbiotic composition comprises less than 0.5% w / w tryptamine relative to ILA.

103. The method of any one of claims 81-102, wherein the postbiotic composition comprises less than 0.5% w / w indole-3-acetamide relative to ILA.

104. The method of any one of claims 81-103, wherein the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising:1 g / L to 50 g / L tryptophan;100 g / L to 200 g / L glucose; manganese; and10 g / L to 125 g / L of one or more nitrogen sources.

105. The method of any one of claims 81-104, wherein the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising:1 g / L to 50 g / L tryptophan;1 g / L to 100 g / L glucose; manganese; and125 g / L to 200 g / L of one or more nitrogen sources.

106. The method of claim 104 or 105, wherein the one or more nitrogen sources comprise yeast extract or peptones.PCT / US25 / 16818 21 February 2025 (21.02.2025)107. The method of any one of claims 81-103, wherein: the first fermentate is produced by incubating the bacterial strain under microaerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose; and one or more of polysorbate 80, manganese, and vitamin B 12.

108. The method of any one of claims 81-103, wherein: the first fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, 4 g / L to 200 g / L glucose, and one or more of polysorbate 80, manganese, and vitamin B12; and the second fermentate is produced by incubating the bacterial strain under aerobic conditions in a fermentation medium comprising 1 g / L to 50 g / L tryptophan, less than 4 g / L glucose, no detectable polysorbate 80, no detectable manganese, and no detectable vitamin B12.

109. The method of any one of claims 81-108, wherein the second fermentate comprises more lAld than the first fermentate.

110. The method of any one of claims 81-109, wherein the first fermentate comprises more ILA than the second fermentate.

111. The method of any one of claims 81-103, wherein the first fermentation medium and / or the second fermentation medium comprises tryptophan at a concentration of 1 to 12 g / L.

112. The method of any one of claims 81-103 or 111, wherein the first fermentation medium comprises glucose at a concentration of at least 4 g / L.

113. The method of any one of claims 81-103 or 111-112, wherein the first fermentation medium comprises glucose at a concentration of at least 4 to 15 g / L.PCT / US25 / 16818 21 February 2025 (21.02.2025)114. The method of any one of claims 81-103 or 111-113, wherein the second fermentation medium comprises glucose at a concentration of less than 4 g / L.

115. The method of any one of claims 81-103 or 111-114, wherein the first fermentation medium comprises polysorbate 80, manganese, and / or vitamin B12.

116. The method of any one of claims 81-103 or 111-115, wherein the second fermentation medium does not comprise polysorbate 80, does not comprise manganese, and does not comprise vitamin B12.

117. The method of any one of claims 81-116, wherein the fermenting of (i) and / or the fermenting of (ii) is for 2 to 7 days.

118. The method of any one of claims 81-117, wherein the fermenting of (i) and / or the fermenting of (ii) is under aerobic or microaerobic conditions.

119. The method of any one of claims 81-118, further comprising separating bacterial cells from the first fermentate and / or second fermentate before combining the first and second fermentates.

120. The method of any one of claims 81-119, further comprising heat killing bacterial strains in the first fermentate and / or second fermentate before combining the first and second fermentates.

121. The method of any one of claims 81-120, further comprising concentrating the first fermentate and / or second fermentate before combining the first and second fermentates.

122. The method of any one of claims 81-121, further comprising drying the first fermentate and / or second fermentate before combining the first and second fermentates.

123. A postbio tic composition produced by the method of any one of claims 81-122.

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