WO2021138599A1 - Cosmetic compositions - Google Patents

Abstract

Described herein are methods and compositions for the prevention or reduction of a skin condition in a subject. In some instances, compositions described herein are provided for cosmetic purposes, such as improvement in irregularities in the texture and appearance of the skin. Provided herein are compositions for moisturization of skin, wherein the composition comprises: at least one species of gram-negative bacteria, wherein the at least one species of gram-negative bacteria is purified and viable; and a humectant or emollient, wherein the composition is in a topical dosage form.

Description

COSMETIC COMPOSITIONS

CROSS-REFERENCE

[0001] This application claims benefit of U.S. Provisional Application No. 62/956,861, filed on January 3, 2020, which is incorporated herein by reference in its entirety.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on December 30, 2020, is named 53654-710_601_SL.txt and is 2,056 bytes in size.

BACKGROUND

[0003] Dysbiosis of the skin microbiome is associated with irregularities in the texture and appearance of the skin. Symptoms of irregularities are often attributed to loss of commensal diversity or microbiota dysfunction. As such, there is a need for compositions for improvement of the texture and appearance of the skin condition of a subject.

BRIEF SUMMARY

[0004] Provided herein are compositions for moisturization of skin, wherein the composition comprises: at least one species of gram-negative bacteria, wherein the at least one species of gram-negative bacteria is purified and viable; and a humectant or emollient, wherein the composition is in a topical dosage form. Further provided herein are compositions wherein the at least one species of gram-negative bacteria is isolated from skin of a donor. Further provided herein are compositions wherein the at least one species of gram-negative bacteria is a commensal organism. Further provided herein are compositions wherein the humectant is glycerin, propylene glycol, dipropylene glycol, polyethylene glycol, butylene glycol, hexane triol. Further provided herein are compositions wherein the emollient is urea, ammonium lactate, vitamin A, vitamin D, or vitamin E. Further provided herein are compositions further comprising the humectant and the emollient.

[0005] Provided herein are compositions for cleansing of skin, comprising: at least one species of gram-negative bacteria, wherein the at least one species of gram-negative bacteria is purified and viable; and a detergent, wherein the composition is in a topical dosage form. Further provided herein are compositions wherein the at least one species of gram-negative bacteria is isolated from skin of a donor. Further provided herein are compositions wherein the at least one species of gram-negative bacteria is a commensal organism. Further provided herein are compositions wherein the detergent comprises sodium laurel sulfate, sodium tallowate, sodium cocoate, or sodium palmate.

[0006] Provided herein are compositions for reduction in sunlight exposure or damage from sunlight exposure comprising: at least one species of gram-negative bacteria, wherein the at least one species of gram-negative bacteria is purified and viable; and an agent to reduce sunlight exposure or damage from sunlight exposure, wherein the composition is in a topical dosage form. Further provided herein are compositions wherein the at least one species of gram negative bacteria is isolated from skin of a donor. Further provided herein are compositions wherein the at least one species of gram-negative bacteria is a commensal organism. Further provided herein are compositions wherein the agent to reduce sunlight exposure is octyl methoxyl cinnamate or butyl methoxy benzoylmethane. Further provided herein are compositions further comprising an emulsifier. Further provided herein are compositions wherein the emulsifier is glycerol stearate, polyglyceryl-3 hydroxylauryl ether, polysorbate 60, or sorbitol polyethylene glycol. Further provided herein are compositions further comprising an antioxidant. Further provided herein are compositions wherein the antioxidant is vitamin C, vitamin E, vitamin A, or a tocopherol. Further provided herein are compositions wherein the at least one species of gram-negative bacteria is present in an amount of up to about 10² CFUs. Further provided herein are compositions wherein the at least one species of gram-negative bacteria comprises Roseomonas mucosa, Acinetobacter radioresistens, Moraxella osloensis, Pantoea septica, Pseudomonas luteola, Pseudomonas aeruginosa , or Pseudomonas oryzihabitans. Further provided herein are compositions wherein the at least one species of gram-negative bacteria comprises a species of Roseomonas. Further provided herein are compositions wherein the species of Roseomonas is Roseomonas mucosa. Further provided herein are compositions wherein the at least one species of gram-negative bacteria are isolated. [0007] Provided herein are compositions as described herein, wherein the topical dosage form is a solid, semisolid, cream, lotion, gel, foam, ointment, powder, or liquid. Further provided herein are composition wherein the topical dosage form comprises a water-based formulation [0008] Provided herein are methods of preventing or reducing a skin condition in a subject, comprising topically administering to a subject the composition as described herein. Further provided herein are methods of preventing or reducing a skin condition in a subject, wherein the skin condition comprises fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof. Further provided herein are methods wherein the subject is an infant, child, or adult. Further provided herein are methods of preventing or reducing a skin condition in a subject, wherein the composition is administered to the subject at least two times per a week. Further provided herein are methods of preventing or reducing a skin condition in a subject, wherein the composition is administered to the subject every other day over a week. Further provided herein are methods of preventing or reducing a skin condition in a subject, wherein the composition is administered to the subject daily. Further provided herein are methods of preventing or reducing a skin condition in a subject, wherein the composition is administered to the subject multiple times a day. Further provided herein are methods of preventing or reducing a skin condition in a subject, wherein the composition is applied to a bandage for topical administration to the subject.

[0009] Provided herein are methods of preventing or reducing a skin condition in a subject, comprising topically administering at least one strain of Roseomonas mucosa to a subject, wherein the Roseomonas mucosa is purified and viable , and wherein the skin condition comprises: wherein the skin condition is fine line occurrence, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof. Further provided herein are methods wherein the subject is an infant, child, or adult. Further provided herein are methods wherein the composition is administered to the subject at least two times per a week. Further provided herein are methods wherein the composition is administered to the subject every other day over a week. Further provided herein are methods wherein the composition is administered to the subject daily. Further provided herein are methods wherein the composition is administered to the subject multiple times a day. Further provided herein are methods wherein the composition is administered to the subject 2, 3, 4, 5, 6 or more times a day. Further provided herein are methods wherein the Roseomonas mucosa is present in an amount of up to about 10² CFUs. Further provided herein are methods wherein the Roseomonas mucosa is in a topical dosage form. Further provided herein are methods wherein the topical dosage form is a solid, semisolid, cream, lotion, gel, foam, ointment, powder, or liquid. Further provided herein are methods wherein the administering provides for a beneficial shift in a cosmetic skin assessment score. Further provided herein are methods wherein the cosmetic skin assessment score is measured using SCORAD criteria, EASI criteria, or comparison to a standard skin model.

BRIEF DESCRIPTION OF THE DRAWINGS [0010] FIG. 1A depicts a route of administration for bacteria via topical administration or oral administration.

[0011] FIG. IB depicts a route of administration for bacteria via rectal administration.

[0012] FIG. 2A depicts a treatment protocol for administration of bacterial compositions.

[0013] FIG. 2B depicts improvement by SCORAD index in patient cohorts receiving treatment with bacterial compositions at a constant dose (10e5) or increasing dosage (Dose Esc).

[0014] FIG. 2C depicts improvement by EASI index in patient cohorts receiving treatment with bacterial compositions at a constant dose (10e5) or increasing dosage (Dose Esc).

DETAILED DESCRIPTION

[0015] Provided herein are compositions and methods for improvement of a skin condition of a subject, and/or reduction or prevention of a skin condition associated with a cosmetic irregularity. Such compositions may include one or more bacteria isolated from a donor subject, or one or more metabolites isolated from the donor bacteria. Compositions described here provide for improvement of the state of the skin condition in a subject. Further described herein are (1) microorganisms for improvement of a skin condition; (2) metabolites for improvement of a skin condition; (3) formulations; (4) administration schedules; (5) kits; and (6) assessment. Such compositions may improve a cosmetic feature of the skin and prevent or reduce a cosmetic irregularity of the skin. In some instances, the bacteria and/or metabolite(s) is provided for prevention or reduction of a cosmetic feature of the skin, for example, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

[0016] Throughout this disclosure, various embodiments are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of any embodiments. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range to the tenth of the unit of the lower limit unless the context clearly dictates otherwise. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual values within that range, for example, 1.1, 2, 2.3, 5, and 5.9. This applies regardless of the breadth of the range. The upper and lower limits of these intervening ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included, unless the context clearly dictates otherwise.

[0017] The terminology used herein is for the purpose of describing particular instances only and is not intended to be limiting of any embodiment. As used herein, the singular forms “a,” “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

[0018] Unless specifically stated or obvious from context, as used herein, the term “about” in reference to a number or range of numbers is understood to mean the stated number and numbers +/- 10% thereof, or 10% below the lower listed limit and 10% above the higher listed limit for the values listed for a range.

Microorganisms

[0019] Provided herein are compositions and methods improvement of a skin condition of a subject, and/or reduction or prevention of a skin condition associated with a cosmetic irregularity. Such skin conditions include reduction or prevention of a cosmetic irregularity or improvement of a measurable feature of the skin subsequent to administration of the composition. Such compositions may include isolated and/or purified bacteria and combinations of bacteria from intact human skin, or propagated from such bacteria. These bacteria can function as a healthy microbiota or promote growth of resident microbiome when administered to a subject for improvement of a skin condition associated with a cosmetic irregularity. The compositions provided may ameliorate, improve, lessen, or diminish the symptoms of cosmetic irregularities of the skin for which improvement is desired. Exemplary cosmetic irregularities include, without limitation, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

[0020] Provided herein are compositions comprising bacteria isolated from a donor subject that does not have a skin condition associated with a cosmetic irregularity. In some instances, the donor subject does not demonstrate a pathological condition of the skin. In some instances, the donor subject does have atopic dermatitis of the skin. The bacteria may be isolated from the skin of the donor subject directly, or propagated in vitro using techniques for culturing bacteria. [0021] Provided herein are genera, species, strains, and combinations of strains or species, originally found within the human skin microbiota of a donor subject. Such species/strains may be selected for their ability to significantly reduce the rate of skin pathogen replication. These species/strains provide a safe and effective means for modulating growth, replication, and disease severity of bacterial pathogens. Moreover, the compositions provided herein exclude pathogenic bacteria. As such, bacteria described herein for use in compositions may be non- pathogenic when administered to the skin of the subject, for example, an immunocompetent subject. Where the bacteria do not cause infection when administered to intact human skin, no pathogenesis is expected to be observed following application. Bacteria obtained from a donor subject may be isolated from the skin of various parts of the donor subject’s body, for example, the face, forearm, antecubital fossa, and neck.

[0022] Compositions described herein, when administered to a subject having a skin condition associated with a cosmetic irregularity, reduce the growth rate of a specific pathogen present in the subject, for example, S. aureus. Bacteria with the capacity to durably reduce S. aureus in the skin can be identified using a methodology for estimating an Ecological Control Factor (ECF) for constituents within the human microbiota. The ECF can be determined by assessing the antagonistic activity of a given commensal strain or combination of strains towards a given pathogen using an in vitro assay, resulting in observed levels of ecological control at various concentrations of the added commensal strains. The ECF for a commensal strain or combination of strains is similar to the minimal inhibitory concentration (MIC) assessment that is employed in the assessment of antibiotics. The ECF can be used to assess and rank the relative potencies of commensal strains and combinations of strains by the ability to antagonize skin pathogens. The ECF of a commensal strain or combination of 20 strains can be calculated by assessing the concentration of that composition that can mediate a given percentage of inhibition (e.g., at least 10%, at least 20%, at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%) of a target pathogen in an in vitro assay.

[0023] Bacteria compositions provided herein may stimulate human keratinocytes. Such stimulation may occur in vivo and/or in vitro. Bacteria can stimulate keratinocytes by increasing the transcription of the mRNA of immune mediators or molecules involved in epithelial barrier function including, for example, increasing production of an mRNA-encoding IL-Ib, an mRNA- encoding defensin beta 4, an mRNA-encoding Cyp27bl, an mRNA-encoding a vitamin D receptor, an mRNA-encoding occludin, an mRNA-encoding claudin 1, and/or an mRNA- encoding fillagrin. Bacterial compositions described herein may induce cytokine expression from human cells. Exemplary impacted human cells include, without limitation, the cells of the skin, such as fibroblasts and keratinocytes. Exemplary induced cytokines include, without limitation, an interleukin (IL), such as IL-6 and IL-Ib.

[0024] In some embodiments, bacteria from only a single genus are included in a composition for improvement of a skin condition of a subject. In alternative embodiments, combinations of genera are included in a composition for improvement of a skin condition associated with a cosmetic irregularity. In further embodiments, the composition comprises bacteria that are viable. Compositions described herein may include, for example, 1, 2, 3, 4, or 5 genera of bacteria.

[0025] In some embodiments, bacteria described herein for improvement of a skin condition of a subject are gram-positive bacteria or gram-negative bacteria. In some embodiments, the bacteria are from a healthy donor. In some embodiments, the bacteria are from a donor. Exemplary gram-positive bacteria include, without limitation, a Staphylococcus species including, without limitation, Staphylococcus epidermidis, Staphylococcus cohnii, and Staphylococcus hominis. Exemplary gram-negative bacteria include, without limitation, Proteobacteria, Acetobacteraceae, Spirochaetaceae, Enter obacteriales, Fusobacterium polymorphum , and Selenomaonadales . Exemplary genera of gram-negative bacteria additionally include Pseudomonas , Pantoea, Moraxella , Roseomonas, Vitreoscilla, and Methylobacteria spp. The gram-negative bacteria may be diplococci, coccobacilli, cocci, or bacilli. Additional bacteria for improvement of a skin condition associated with a cosmetic irregularity include, without limitation, Lactobacillus casei var. rhamnosus, Bifidobacterium animalis subsp lactis. Bifidobacterium longum, Lactobacillus plantarum, and Lactobacillus johnsonii. In some instances, the bacteria is provided for prevention or reduction of a cosmetic feature of the skin, for example, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

[0026] In some embodiments, a composition provided herein comprises a viable species of Roseomonas. In some embodiments, a composition provided herein comprises a viable species of Pseudomonas. In some embodiments, a composition provided herein comprises a viable species of Roseomonas and viable species of Pseudomonas. In some instances, the bacteria is provided for prevention or reduction of a cosmetic feature of the skin, for example, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof. Compositions described herein may include one or more of a species of the Roseomonas genus for improvement of a skin condition. In some instances, the bacteria is provided for prevention or reduction of a cosmetic feature of the skin, for example, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof. Exemplary species of the Roseomonas genus include, without limitation, Roseomonas aerilata , Roseomonas aerophila , Roseomonas aestuarii, Roseomonas alkaliterrae , Roseomonas aquatic , Roseomonas cervicalis , Roseomonas fauriae , Roseomonas frigidaquae , Roseomonas gilardii , Roseomonas lacus , Roseomonas ludipueritiae , Roseomonas mucosa , Roseomonas pecuniae , Roseomonas rhizosphaerae , Roseomonas riguiloci , Roseomonas rosea , Roseomonas soli , Roseomonas stagni, Roseomonas terrae , and Roseomonas vinacea. The bacteria may be viable. The bacteria may be isolated and/or purified. The bacteria may be isolated from a subject not having the skin condition associated with a cosmetic irregularity which is sought to be treated.

[0027] Compositions described herein may include one or more of a species of the Pseudomonas genus for improvement of a skin condition. Exemplary species of the Pseudomonas genus include, without limitation, Pseudomonas aeruginosa , Pseudomonas luteola , and Pseudomonas oryzihabitans. The bacteria may be viable. The bacteria may be isolated and/or purified. The bacteria may be isolated from a subject not having the skin condition associated with a cosmetic irregularity which is sought to be treated. Compositions described herein may include one or more of a species of the Pantoea genus for improvement of a skin condition. Exemplary species of the Pantoea genus include, without limitation, Pantoea septica. The bacteria may be viable. The bacteria may be isolated and/or purified.

Compositions described herein may include one or more of a species of the Moraxella genus for improvement of a skin condition. Exemplary species of the Moraxella genus include, without limitation, Moraxella osloensis. Compositions described herein may include one or more of a species of the Vitreoscilla genus for improvement of a skin condition. Exemplary species of the Vitreoscilla genus include, without limitation, Vitreoscilla filiformis, Vitreoscilla beggiatoides, and Vitreoscilla stercoraria. The bacteria may be viable. The bacteria may be isolated and/or purified. The bacteria may be isolated from a subject not having the skin condition associated with a cosmetic irregularity which is sought to be treated. In some instances, the bacteria is provided for prevention or reduction of a cosmetic feature of the skin, for example, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

[0028] In some embodiments, bacteria of a single species or a single strain are included in compositions disclosed herein. In some embodiments, combinations of species bacteria are included in compositions for use in disclosed methods. Thus, in some embodiments described herein, a composition includes 1, 2, 3, 4, or 5 species of bacteria. In some embodiments, a composition provided herein includes multiple viable Roseomonas mucosa strains from one or more healthy donor subjects. In some embodiments, a composition provided herein includes multiple viable Pseudomonas aeruginosa strains from one or more healthy donor subjects. In some embodiments, a composition provided herein includes multiple viable Roseomonas mucosa strains from one or more donor subjects. In some embodiments, a composition provided herein includes multiple viable Pseudomonas aeruginosa strains from one or more donor subjects.

[0029] Compositions provided herein may include two types of bacteria or more than two types of bacteria. The compositions may include 1 to 15, 2 to 12, 2 to 10, or 2 to 5 different species of bacteria. The compositions may include 1 to 15, 2 to 12, 2 to 10, or 2 to 5 different strains of bacteria. The compositions may include 1 to 15, 2 to 12, 2 to 10, or 2 to 5 different strains of the same species of bacteria. The compositions may include 1, 2, 3, 4 or 5 different strains of the same species of bacteria. The compositions may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more different species of bacteria. In some embodiments, compositions include 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, at least 20, at least 30, at least 40, at least 50, or greater than 50 types of bacteria, as defined by genus, species, or operational taxonomic unit (OTU).

[0030] Compositions provided herein may comprise one or more bacteria having a percent identity based on 16S rRNA bacterial genetic sequence to a reference strain. The comparison of the 16S rRNA bacterial genetic sequence allows a strain to be identified by comparing sequences with known bacterial DNA sequences using BLAST

(https://blast.ncbi.nlm.nih.gov/Blast.cgi). The level of identity in relation to a nucleotide sequence may be determined for at least 20 contiguous nucleotides, for at least 30 contiguous nucleotides, for at least at least 40 contiguous nucleotides, for at least 50 contiguous nucleotides, for at least 60 contiguous nucleotides, or for at least 100 contiguous nucleotides. Preferably, the level of identity in relation to a nucleotide sequence is determined for the entire sequence searched. In some instances, the percent identity is based on whole genome comparison, 16s rRNA, or a semi-conserved hypervariable region, such as the V4 region. Percent identity may be at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% to a reference 16S rRNA bacterial genetic sequence. In some instances, a stain enriched by methods described herein has at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the whole genome, 16s rRNA or V4 hypervariable region of a strain listed in Table 1.

Table 1

[0031] Bacteria described herein may be transformed with a heterologous nucleic acid, such as in the form of a plasmid. For example, the plasmid may comprise an expression vector encoding for protein of interest. Such a mechanism provides a means for introduction of exogenous DNA can be introduced bacterial cells with standard techniques, such as electroporation or calcium phosphate-mediated transfection.

[0032] In some embodiments, the heterologous nucleic acid is included in a plasmid. A plasmid generally contains multiple genetic elements positionally and sequentially oriented with other necessary genetic elements, such that the nucleic acid in a nucleic acid cassette can be transcribed and when necessary translated in the transfected cells. Plasmids can include nucleic acids that are derived from DNA via a plasmid vector, cosmids, or phagemids, where one or more heterologous nucleic acid can be inserted. The heterologous nucleic acid can encode a protein of interest, which can be operably-linked to a promoter for expression of the bacteria. [0033] Plasmids generally contain one or more unique restriction sites. In addition, a plasmid can confer well-defined phenotypes on the host organism, which can be selectable or readily- detected, for example, drug resistance. Thus, the plasmid can include an expression cassette, where a polypeptide is encoded. Expression can include the efficient transcription of an inserted gene, nucleic acid sequence, or nucleic acid cassette with the plasmid. [0034] In some embodiments, when a circular plasmid is transferred into a bacterial cell, the plasmid can be an autonomously replicating, extra-chromosomal DNA molecule, distinct from the normal bacterial genome and non-essential for bacterial cell survival under non-selective conditions. Persistent expression can refer to introduction of genes into the cell together with genetic elements which enable episomal (extra-chromosomal) replication and/or maintenance of the genetic material in the cell. Persistent expression can lead to apparently stable transformation of the cell without the integration of the novel genetic material into the chromosome of the host cell. A plasmid can also introduce genetic material into chromosomes of the targeted cell. Gene expression after stable introduction can permanently alter the characteristics of the cell and cell progeny by replication leading to stable transformation.

[0035] Methods for producing bacterial strains for incorporation in a composition described herein optionally include processing steps of organism banking, organism production, and preservation. For organism banking, strains of bacteria can be isolated directly from a specimen, for example, from human skin or a banked stock. Bacteria can be cultured on a nutrient agar or broth that supports growth to generate viable biomass. The cultured biomass can be preserved in multiple aliquots for long-term storage. Bacteria may be isolated directly from the skin of a human donor subject. Generally, the human donor subject does not have a skin pathology, such as atopic dermatitis. Bacteria can also be isolated from other sources including, for example, commercial sources or environmental samples.

Microbial Metabolites

[0036] Certain strains or species of bacteria have been reported to have a beneficial effect on skin conditions, such as those associated with a cosmetic irregularity. The beneficial effect of such bacteria may be attributed to the metabolic profile of the bacteria, i.e. metabolite produced by the bacteria. In some instances, bacteria can produce lipids that have a beneficial effect on restoring barrier function and regulating immune response.

[0037] Provided herein are compositions and methods improvement of a skin condition of a subject, and/or reduction or prevention of a skin condition associated with a cosmetic irregularity, relating to the use of one or more metabolites produced by bacteria from a healthy donor subject. The one or more metabolites may alleviate the skin condition associated with a cosmetic irregularity and may be those which are increased in production from a species of bacteria present on the skin of a healthy donor. Provided herein are compositions and methods improvement of a skin condition of a subject, and/or reduction or prevention of a skin condition associated with a cosmetic irregularity, relating to the use of one or more metabolites produced by bacteria from a donor subject. The one or more metabolites may alleviate the skin condition associated with a cosmetic irregularity and may be those which are increased in production from a species of bacteria present on the skin of a donor. The one or more metabolites may be part of a combination therapy also including a species of bacteria obtained or derived from a donor subject that does not have the skin condition associated with a cosmetic irregularity. Metabolites described herein for improvement of a skin condition include, without limitation, those listed in Table 2. Provided herein are methods for improvement of a skin condition in a subject comprising administering to a subject in need thereof a composition comprising metabolites listed in Table 2, optionally in addition to administration of one or more species of bacteria described herein for improvement of the skin condition in a subject. Exemplary skin conditions for use of compositions described herein include, without limitation, acne, fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, scaly patches, roughness, acne, scars, irregular pigmentation, sun spots, liver spots, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, tactile smoothness, suppleness, glow, visual smoothness, and radiance.

Table 2. Metabolites

Formulations

[0038] Compositions and described herein may be formulated for topical, oral, or rectal administration. FIG. 1A depicts a route of topical administration for bacteria 101 or a route of oral administration for bacteria 102. FIG. IB depicts a route of rectal administration for bacteria 103. Exemplary topical dosage forms include, without limitation, shampoos, conditioners, skin cleansers, creams, lotions, gels, ointments, pastes, powders, and sterile aqueous solutions or suspensions. Additional dosage forms for compositions described herein include, without limitation, moisturizers, perfumes, lipsticks, fingernail polishes, eye and facial makeup preparations, permanent waves, hair colors, toothpastes, and deodorants.

[0039] In some embodiments, a composition described herein comprises an aqueous carrier, and is applied as a spray to the skin. In some embodiments, a composition described herein comprises a water-based formulation. Shampoos are liquid preparations containing detergent or soap for cleaning the hair. A shampoo, as described herein, is optionally sulfate-free. Conditioners are liquids applied to the hair to improve the condition. Conditioner is applied, without limitation, after application and removal of a shampoo or without prior application of a shampoo. Skin cleansers are compositions used to cleanse the skin. Skin cleansers are, without limitation, formulated for application to the face, for application to the entire body, or for application to a specific part of the body, for example, the hands or the feet. Creams are viscous liquids or semisolid emulsions, either oil-in-water or water-in-oil. Cream bases are water- washable, and contain an oil phase, an emulsifier, and an aqueous phase. The oil or “internal” phase generally contains of petrolatum and a fatty alcohol, such as acetyl or stearyl alcohol. In some embodiments, a composition described herein, the aqueous phase exceeds the oil phase in volume, and optionally contains a humectant. In some embodiments described herein, the emulsifier in a cream formulation is a nonionic, anionic, cationic, or amphoteric surfactant. In some embodiments described herein, the lotion comprises a preparation to be applied to the skin surface without friction. Lotions are typically liquid or semiliquid preparations in which particles are present in a water or alcohol base. In some embodiments described herein, lotions are suspensions of solids or a liquid oily emulsion of the oil-in-water type. In some embodiments described herein, lotions are used for treating large body areas, because of the ease of applying a more fluid composition. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the active agent in contact with the skin, e.g., methylcellulose, sodium carboxymethyl-cellulose, or the like. Gels described herein may be semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the carrier liquid, which in some embodiments described herein are aqueous, contain an alcohol, or are hydrophobic. In some embodiments, organic macromolecules, including gelling agents, are crosslinked acrylic acid polymers, e.g., carboxypolyalkylenes (CARBOPOL®). Non-limiting examples of gels include hydrophilic polymers, such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers, and polyvinylalcohol; cellulosic polymers, such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums, such as tragacanth and xanthan gum; sodium alginate; and gelatin. To prepare a uniform gel, dispersing agents, such as alcohol or glycerin are optionally added. Alternatively, the gelling agent is dispersed by trituration, mechanical mixing or stirring, or combinations thereof. Ointments describe herein may be semisolid preparations that are typically based on petrolatum or other petroleum derivatives. The specific ointment base to be used may be one that will provide for a number of desirable characteristics, e.g., emolliency or the like. An ointment base is generally inert, stable, non-irritating, and non sensitizing. Ointment bases may be oleaginous bases, emulsifiable bases, emulsion bases, or water-soluble bases. Pastes are semisolid dosage forms in which the active agent is suspended in a suitable base, and are also of use. Depending on the nature of the base, pastes are divided between fatty pastes or those made from single-phase aqueous gels. In some embodiments described herein, the base in a fatty paste is petrolatum or hydrophilic petrolatum or the like. In some embodiments described herein, the pastes made from single-phase aqueous gels incorporate carboxymethylcellulose or the like as a base. In some embodiments described herein, powders are packaged for reconstitution in any topical dosage form described herein, or for direct topical application. In some embodiments described herein, a powder comprises lyophilized bacteria prepared for preservation as described herein. In some embodiments described herein, a powder comprises additional carriers to improve, for example, flowability, bulking, or aesthetic features. In some embodiments described herein, additional carriers include, without limitation, silica, starch, talc, dimethicone, zirconium silicate, zinc oxide, titanium dioxide, kaolin, or magnesium carbohydrate.

[0040] Solutions may be homogeneous mixtures prepared by dissolving one or more chemical substances (solutes) in a liquid such that the molecules of the dissolved substance are dispersed among those of the solvent. In some embodiments described herein, a solution contains other pharmaceutically acceptable chemicals to buffer, stabilize, or preserve the solute. Common examples of solvents used in preparing topical solutions are ethanol, water, propylene glycol or any other-acceptable vehicles. In some embodiments described herein, these are applied in any manner, such as spraying them on the skin, painting them on the skin, or wetting a bandage with the solution.

[0041] In some embodiments, a topical composition described herein is any form suitable for application to the body surface including, for example, as a cream, lotion, spray, solution, gel, foam, ointment, powder, paste, plaster, paint, bioadhesive, bandage, spray, suspension, and containing liposomes, micelles, and/or microspheres. In some embodiments, a topical composition described herein is used in combination with an occlusive overlayer so that moisture evaporating from the body surface can be maintained within the formulation upon application to the body surface and thereafter. In some embodiments a cream, lotion, gel, ointment, paste, powder, or the like is spread on the affected surface.

[0042] In some embodiments a composition provided herein is applied directly to a target location in a subject, for example in a topical preparation, or as a part of a dressing or a bandage. In some embodiments a composition provided herein is formulated as a unit dosage, for administration by any device for administration to the skin. The unit dosage can be a reservoir of the active agent in a carrier, for example an adhesive carrier capable of adhering to the skin for a desired period of time, such as at least a day or more.

[0043] Exemplary oral dosage forms include, without limitation, a tablet, lozenge, pastille capsule, tab, granules, powder, liquid, emulsion, suspension, and syrup. Exemplary rectal dosage forms include, without limitation, a suppository, and enema solution, rectal foam, or rectal gel.

[0044] In some embodiments a composition provided herein includes a carrier. In some embodiments, compositions include additional active and/or inactive materials, which is prepared as single dosage unit or in a multi-dose format. Compositions described herein may include a carrier that comprises one or more of a buffering agent, a preservative, a stabilizer, a binder, a compaction agent, a lubricant, a dispersion enhancer, and/or a coloring agent. Non limiting examples of suitable buffering agents include sodium citrate, magnesium carbonate, magnesium bicarbonate, calcium carbonate, and calcium bicarbonate. Non-limiting examples of suitable preservatives include antioxidants, such as alpha tocopherol and ascorbate, parabens, chlorobutanol, and phenol. Non -limiting examples of suitable binders include sucrose, starches, pregelatinized starches, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethyl cellulose, polyacrylamides, polyvinyloxoazolidone, polyvinylalcohols, C 12-08 fatty acid alcohol, polyethylene glycol, polyols, saccharides, oligosaccharides, and combinations thereof. Non-limiting examples of suitable lubricants include magnesium stearate, calcium stearate, zinc stearate, hydrogenated vegetable oils, sterotex, polyoxyethylene monostearate, talc, polyethyleneglycol, sodium benzoate, sodium lauryl sulfate, magnesium lauryl sulfate, and light mineral oil. A pH buffering agent(s) can, if employed and when dissolved in an aqueous component of the composition, provide a pH in the range of 5 to 7 (5 e.g. about pH 5.5).

[0045] Compositions described herein may include a carrier that comprises other ingredients including, for example, ingredients that sustain growth of the bacteria. In some embodiments, a composition provided herein includes a nutrient. In some embodiments, compositions include at least one carbohydrate or saccharide. In some embodiments, a carbohydrate is a monosaccharide, a disaccharide, tri saccharide, oligosaccharide, or polysaccharide. Non-limiting examples of carbohydrates include glucose, sucrose, galactose, mannose, ribose, arabinose, xylose, fructose, maltose, cellobiose, lactose, raffmose, stachyose, starch, glycogen, and cellulose. In some embodiments, carbohydrates contain modified saccharide unit, including, for example, 2'-deoxyribose in which a hydroxyl group is removed, 2'-fluororibose in which a hydroxyl group is replaced with a fluorine, and or N-acetylglucosamine, a nitrogen-containing form of glucose (e.g., 2'-fluororibose, deoxyribose, and hexose). Carbohydrates exist in many different forms, for example, without exclusion, conformers, cyclic forms, acyclic forms, stereoisomers, tautomers, anomers, and isomers.

[0046] Compositions described herein may include a carrier that comprises one or more lipids.

In some embodiments a lipid comprises fats, oils, triglycerides, cholesterol, phospholipids, and fatty acids. Fats, oils, and fatty acids can be saturated, unsaturated (cis or trans), or partially unsaturated (cis or trans). Non-limiting examples of fatty acids include lauric acid (12:0), myristic acid (14:0), palmitic acid (16:0), palmitoleic acid (16:1), margaric acid (17:0), heptadecenoic acid (17:1), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2), linolenic acid, a-linolenic acid, and g-linolenic acid.

[0047] Compositions described herein may include a carrier that comprises at least one supplemental mineral or mineral source. Non-limiting examples of minerals include chloride, sodium, calcium, iron, chromium, copper, iodine, zinc, magnesium, manganese, molybdenum, phosphorus, potassium, and selenium. Suitable forms of any of the foregoing minerals include soluble mineral salts, slightly soluble mineral salts, insoluble mineral salts, chelated minerals, mineral complexes, non-reactive minerals, such as carbonyl minerals, and reduced minerals, and combinations thereof. In some embodiments, compositions include at least one supplemental vitamin. Supplemental vitamins can be fat-soluble or water-soluble. Non-limiting examples of vitamins include vitamin C, vitamin A, vitamin E, vitamin B 12, vitamin K, riboflavin, niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine, pantothenic acid, and biotin. Suitable forms of any of the foregoing are salts of the vitamin, derivatives of the vitamin, compounds having the same or similar activity of the vitamin, and metabolites of the vitamin.

[0048] Various other additives can be included in the compositions. Non-limiting examples of additives include antioxidants, astringents, perfumes, preservatives, emollients, pigments, detergents, dyes, humectants, propellants, and sunscreen agents, as well as other classes of materials whose presence can be pharmaceutically or otherwise desirable. Non-limiting examples of optional additives include preservatives, such as sorbate; detergents, such as sodium laurel sulfate, sodium tallowate, sodium cocoate, or sodium palmate (or alternative salts of the foregoing, such as potassium or magnesium); solvents, such as isopropanol and propylene glycol; astringents, such as menthol and ethanol; emollients, such as polyalkylene methyl glucoside, urea, ammonium lactate, vitamin A, vitamin D, or vitamin E; humectants, such as glycerin, propylene glycol, dipropylene glycol, polyethylene glycol, butylene glycol, hexane triol; emulsifiers, such as glycerol stearate, PEG-100 stearate, polyglyceryl-3 hydroxylauryl ether, and polysorbate 60; sorbitol and other polyhydroxyalcohols, such as polyethylene glycol; sunscreen agents, such as octyl methoxyl cinnamate (Parsol MCX) and butyl methoxy benzoylmethane (Parsol 1789); antioxidants, such as ascorbic acid (vitamin C), a-tocopherol (Vitamin E), b-tocopherol, g-tocopherol, d-tocopherol, e-tocopherol, zi -tocopherol, Cp.- tocopherol, h-tocopherol, and retinol (vitamin A); essential oils, ceramides, essential fatty acids, mineral oils, vegetable oils (e.g., soya bean oil, palm oil, liquid fraction of shea butter, sunflower oil), animal oils (e.g., perhydrosqualene), synthetic oils, silicone oils or waxes (e.g., cyclomethicone and dimethicone), fluorinated oils (generally perfluoropolyethers), fatty alcohols (e.g., cetyl alcohol), and waxes (e.g., beeswax, camauba wax, and paraffin wax); skin- feel modifiers; and thickeners and structurants, such as swelling clays and cross-linked carboxypolyalkylenes.

[0049] Other additives include materials that condition the skin. Such materials can soften the skin by retarding the decrease of water content of the skin and/or protect the skin. Conditioners and moisturizing agents include, for example, pyrrolidine carboxylic acid and amino acids; organic antimicrobial agents, such as triclosan and benzoic acid. Further additives include anti inflammatory agents, such as acetylsalicylic acid and glycyrrhetinic acid; anti-seborrhoeic agents, such as retinoic acid; vasodilators, such as nicotinic acid; inhibitors of melanogenesis, such as kojic acid; and mixtures thereof.

[0050] In some embodiments, compositions described herein include alpha hydroxyacids, alpha ketoacids, polymeric hydroxyacids, moisturizers, collagen, marine extract, and antioxidants, such as ascorbic acid (vitamin C) and a-tocopherol (Vitamin E). Sunscreens can also be included. Additional, components, such as enzymes, herbs, plant extracts, and glandular or animal extracts can be added to the composition. The amounts of these various additives are those conventionally used in the cosmetics field, and range, for example, from about 0.01% to about 20% of the total weight of the topical formulation.

[0051] Compositions described herein optionally include antimicrobial agents, to prevent spoilage upon storage, for example, to inhibit growth of microbes, such as yeasts and molds. Suitable antimicrobial agents are typically selected from the group consisting of the methyl and propyl esters of p-hydroxybenzoic acid (i.e., methyl and propyl paraben), sodium benzoate, sorbic acid, imidurea, and combinations thereof.

[0052] In some embodiments, compositions described herein contain irritation-mitigating additives to reduce or eliminate the possibility of skin irritation or skin damage resulting from the chemical entity to be administered, or other components of the composition. Suitable irritation-mitigating additives include, for example, a-tocopherol; monoamine oxidase inhibitors, particularly phenyl alcohols, such as 2-phenyl- 1 -ethanol; glycerin; salicylates; ascorbates; ionophores, such as monensin; amphiphilic amines; ammonium chloride; N- acetylcysteine; capsaicin; and chloroquine. The irritation-mitigating additive, if present, can be incorporated into the compositions at a concentration effective to mitigate irritation or skin damage, typically representing not more than about 20 wt%, more typically not more than about 5 wt%, of the formulation. Non-limiting examples of suitable pharmacologically-active agents that are incorporated into the present formulations include the following: agents that improve or eradicate pigmented or non-pigmented age spots, keratoses, and wrinkles; local anesthetics and analgesics; corticosteroids; retinoids; and hormones. Some examples of topical pharmacologically-active agents include acyclovir, amphotericins, chlorhexidine, clotrimazole, ketoconazole, econazole, miconazole, metronidazole, minocycline, phenytoin, para-amino benzoic acid esters, octyl methoxycinnamate, octyl salicylate, oxybenzone, dioxybenzone, tocopherol, tocopheryl acetate, zinc pyrithione, diphenhydramine, pramoxine, lidocaine, procaine, crotamiton, hydroquinone and its monomethyl and benzyl ethers, naproxen, ibuprofen, cromolyn, retinol, retinyl palmitate, retinyl acetate, coal tar, griseofulvin, estradiol, hydrocortisone, hydrocortisone 21 -acetate, hydrocortisone 17-valerate, hydrocortisone 17- butyrate, progesterone, betamethasone valerate, betamethasone dipropionate, triamcinolone acetonide, fluocinonide, clobetasol propionate, minoxidil, dipyridamole, diphenylhydantoin, benzoyl peroxide, 5-fluorouracil, tacrolimus, and topical steroids, such as alclometasone, amcinonide, betamethasone, clobetasol, desonide, dexoximetasone, diflorasone, flucinonide, flurandrenolide, halobetasol, halcinonide, hydrocortisone, and/or triamcinolone.

[0053] Although topical formulations, such as creams and salves, are formulated for dermal delivery, in some embodiments described herein, delivery systems include time-release, delayed release, or sustained release delivery systems. Such systems avoid repeated administrations of the compositions, increasing convenience to the subject and the physician. Non-limiting examples of release delivery systems include (a) erosional systems and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer. In some embodiments described herein, the delivery system includes collagen, fibrin, or a membrane extract, such as a basal membrane extract, for example, in which compositions are formulated for administration to the skin. Suitable basement membrane extracts include a biologically active polymerizable extract containing in parts by weight about 60-85% laminin, 5-30% collagen IV, 1-10% nidogen, 1-10% heparan sulfate proteoglycan, and 1-5% entactin. BME can support normal growth and differentiation of various cell types including epithelial cells when cultured. Basal membrane extracts are well known in the art and are commercially available.

[0054] Compositions described herein may comprise a single (unit) dose of bacteria. Compositions described herein may comprise up to about 10², up to about 10³, or up to about 10⁴ colony forming units (cfu) of bacteria or a bacterial strain described herein. Compositions described herein may comprise about 10¹ to about 10⁴ cfu, about 10² to about 10³ cfu, about 10³ to about 10⁴ cfu, 10¹ to about 10¹⁵ cfu, about 10¹ to about 10¹⁴ cfu, about 10¹ to about 10¹³ cfu, about 10¹ to about 10¹² cfu, about 10¹ to about 10¹¹ cfu, about 10¹ to about 10¹⁰ cfu, about 10¹ to about 10⁹ cfu, about 10¹ to about 10⁸ cfu, about 10¹ to about 10⁷ cfu, about 10¹ to about 10⁶ cfu, about 10¹ to about 10⁵ cfu, about 10¹ to about 10⁴ cfu, about 10¹ to about 10³ cfu, about 10¹ to about 10² cfu, about 10² to about 10¹² cfu, about 10² to about 10¹¹ cfu, about 10² to about 10¹⁰ cfu, about 10² to about 10⁹ cfu, about 10² to about 10⁸ cfu, about 10² to about 10⁷ cfu, about 10² to about 10⁶ cfu, about 10³ to about 10¹² cfu, about 10³ to about 10¹¹ cfu, about 10³ to about 10¹⁰ cfu, about 10⁴ to about 10¹² cfu, about 10⁵ to about 10¹² cfu, about 10⁶ to about 10¹² cfu, about

10⁷ to about 10¹² cfu, about 10¹⁰ to about 10¹² cfu, about 10¹¹ to about 10¹² cfu, or about 10⁶ to about 10¹⁰ cfu of bacteria or a bacterial strain described herein. In some embodiments, compositions comprise about 10¹ cfu, about 10² cfu, about 10³ cfu, about 10⁴ cfu, about 10⁵ cfu, about 10⁶ cfu, about 10⁷ cfu, about 10⁸ cfu, about 10⁹ cfu, about 10¹⁰ cfu, about 10¹¹ cfu, about 10¹² cfu, about 10¹³ cfu, about 10¹⁴ cfu, or about 10¹⁵ cfu of bacteria or a bacterial strain described herein.

[0055] Compositions described herein may comprise up to about 10¹ cfu/ml, about 10² cfu/ml, about 10³ cfu/ml, about 10⁴ cfu/ml, about 10⁵ cfu/ml, about 10⁶ cfu/ml, about 10⁷ cfu/ml, about

10⁸ cfu/ml, about 10⁹ cfu/ml, about 10¹⁰ cfu/ml, about 10¹¹ cfu/ml, about 10¹² cfu/ml, about 10¹³ cfu/ml, about 10¹⁴ cfu/ml, or about 10¹⁵ cfu/ml of bacteria or a bacterial strain described herein. Compositions described herein may comprise about 10¹ to about 10⁴ cfu/ml, about 10² to about 10³ cfu/ml, about 10³ to about 10⁴ cfu/ml, 10¹ to about 10¹⁵ cfu/ml, about 10¹ to about 10¹⁴ cfu/ml, about 10¹ to about 10¹³ cfu/ml, about 10¹ to about 10¹² cfu/ml, about 10¹ to about 10¹¹ cfu/ml, about 10¹ to about 10¹⁰ cfu/ml, about 10¹ to about 10⁹ cfu/ml, about 10¹ to about 10⁸ cfu/ml, about 10¹ to about 10⁷ cfu/ml, about 10¹ to about 10⁶ cfu/ml, about 10¹ to about 10⁵ cfu/ml, about 10¹ to about 10⁴ cfu/ml, about 10¹ to about 10³ cfu/ml, about 10¹ to about 10² cfu/ml, about 10² to about 10¹⁵ cfu/ml, about 10² to about 10¹⁴ cfu/ml, about 10² to about 10¹³ cfu/ml, about 10² to about 10¹² cfu/ml, about 10² to about 10¹¹ cfu/ml, about 10² to about 10⁹ cfu/ml, about 10² to about 10⁸ cfu/ml, about 10² to about 10⁷ cfu/ml, about 10² to about 10⁶ cfu/ml, about 10⁵ to about 10¹⁵ cfu/ml, about 10⁵ to about 10¹⁴ cfu/ml, about 10⁵ to about 10¹⁰ cfu/ml, about 10⁶ to about 10¹² cfu/ml, about 10⁷ to about 10¹² cfu/ml, about 10⁸ to about 10¹² cfu/ml, about 10⁹ to about 10¹² cfu/ml, about 10¹⁰ to about 10¹² cfu/ml, about 10¹¹ to about 10¹² cfu/ml, or about 10⁶ to about 10¹⁰ cfu/ml of bacteria or a bacterial strain described herein. In some embodiments, compositions comprise about 10¹ cfu/ml, about 10² cfu/ml, about 10³ cfu/ml, about 10⁴ cfu/ml, about 10⁵ cfu/ml, about 10⁶ cfu/ml, about 10⁷ cfu/ml, about 10⁸ cfu/ml, about 10⁹ cfu/ml, about 10¹⁰ cfu/ml, about 10¹¹ cfu/ml, about 10¹² cfu/ml, about 10¹³ cfu/ml, about 10¹⁴ cfu/ml, or about 10¹⁵ cfu/ml of bacteria or a bacterial strain described herein.

[0056] Compositions described herein may comprise up to about 10¹ cfu/g, about 10² cfu/g, about 10³ cfu/g, about 10⁴ cfu/g, about 10⁵ cfu/g, about 10⁶ cfu/g, about 10⁷ cfu/g, about 10⁸ cfu/g, about 10⁹ cfu/g, or about 10¹⁰ cfu/g of bacteria or a bacterial strain described herein. Compositions described herein may comprise about 10¹ to about 10⁴ cfu/g, about 10² to about 10³ cfu/g, about 10³ to about 10⁴ cfu/g, 10³ to about 10¹¹ cfu/g, about 10³ to about 10¹⁰ cfu/g, about 10³ to about 10⁹ cfu/g, about 10³ to about 10⁸ cfu/g, about 10³ to about 10⁷ cfu/g, about 10³ to about 10⁶ cfu/g, about 10³ to about 10⁵ cfu/g, about 10³ to about 10⁴ cfu/g, about 10⁴ to about 10¹⁵ cfu/g, about 10⁴ to about 10¹⁴ cfu/g, about 10⁴ to about 10¹³ cfu/g, about 10⁴ to about 10¹² cfu/g, about 10⁴ to about 10¹¹ cfu/g, about 10⁴ to about 10¹² cfu/g, about 10⁴ to about 10¹¹ cfu/g, about 10⁵ to about 10¹⁰ cfu/g, about 10⁵ to about 10⁹ cfu/g, about 10⁵ to about 10⁸ cfu/g, about 10⁶ to about 10¹² cfu/g, about 10⁷ to about 10¹² cfu/g, about 10⁸ to about 10¹² cfu/g, about 10⁹ to about 10¹² cfu/g, about 10¹⁰ to about 10¹² cfu/g, about 10¹¹ to about 10¹² cfu/g, or about 10⁶ to about 10¹⁰ cfu/g of bacteria or a bacterial strain described herein. In some embodiments, compositions comprise about 10¹ cfu/g, about 10² cfu/g, about 10³ cfu/g, about 10⁴ cfu/g, about 10⁵ cfu/g, about 10⁶ cfu/g, about 10⁷ cfu/g, about 10⁸ cfu/g, about 10⁹ cfu/g, about 10¹⁰ cfu/g, about 10¹¹ cfu/g, about 10¹² cfu/g, about 10¹³ cfu/g, about 10¹⁴ cfu/g, or about 10¹⁵ cfu/g of bacteria or a bacterial strain described herein.

[0057] In some embodiments, a composition described herein comprises up to about 0.001% by weight, up to about 0.01% by weight, up to about 0.1% by weight, up to about 0.5% by weight, up to about 1% by weight, or up to about 10% by weight of bacteria or bacterial strain described herein. In other embodiments, a composition described herein comprises at least about 0.01 % by weight, at least about 0.05% by weight, at least about 0.1 % by weight, at least about 0.2% by weight, at least about 0.3% by weight, at least about 0.4% by weight, at least about 0.5% by weight, at least about 0.6% by weight, at least about 0.7% by weight, at least about 0.8% by weight, at least about 0.9% by weight, at least about 1.0% by weight, at least about 1.5% by weight, at least about 2.0% by weight, at least about 3.0% by weight, at least about 4.0% by weight, at least about 5.0% by weight, at least about 6.0% by weight, at least about 7.0% by weight, at least about 8.0% by weight, at least about 9.0% by weight, at least about 10.0% by weight, at least about 11.0% by weight, at least about 12.0% by weight, at least about 13.0% by weight, at least about 14.0% by weight, at least about 15.0% by weight, at least about 16.0% by weight, at least about 17.0% by weight, at least about 18.0% by weight, at least about 19.0% by weight, at least about 20.0% by weight, at least about 25.0% by weight, at least about 30.0% by weight, at least about 35.0% by weight, at least about 40.0% by weight, at least about 45.0% by weight, or at least about 50.0% by weight of bacteria or bacterial strain described herein. In some embodiments, compositions include from 0.01 % to 30% by weight, from about 0.01 % to 20% by weight, from 0.01 % to 5% by weight, from 0.1 % to 30% by weight, from 0.1 % to 20% by weight, from 0.1 % to about 15% by weight, from 0.1 % to 10% by weight, from 0.1 % to 5% by weight, from 0.2% to 5% by weight, from 0.3% to 5% by weight, from 0.4% to 5% by weight, from 0.5% to 5% by weight, or from 1% to 5% by weight of bacteria or bacterial strain described herein.

Preservation

[0058] Bacteria described herein may be preserved for banking. In such instances, organisms may be placed into a chemical milieu that protects from freezing (such as by adding cryoprotectants), drying, and/or osmotic shock (such as by adding osmoprotectants), dispensing into multiple (optionally identical) containers to create a uniform bank, and then optionally treating the culture for preservation. Containers are generally impermeable and have closures that assure isolation from the environment. Cryopreservation can be accomplished by freezing a liquid at ultra-low temperatures (e.g., at or below about -70°C.). Dried preservation removes water from the culture by evaporation (in the case of spray drying or cool drying) or by sublimation (e.g., for freeze drying, spray freeze drying). Removal of water (i.e. by lyophilization) is used to improves long-term bacterial composition storage stability at higher temperatures. Strains and/or species can be cultured and preserved individually, or species/strains can be mixed together for banking.

Administration

[0059] Provided herein are methods for preventing or reducing skin conditions by administering compositions described herein to a subject. In some embodiments, the subject is a mammal, e.g., a dog, cat, sheep, pig, mouse, ape, monkey or human. In some embodiments, the subject is a child. In some embodiments, the subject is 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 year(s) of age. In some embodiments, the subject is an adolescent. In some embodiments, the subject is 12, 13, 14, 15, 16, 17, or 18 years of age. In some embodiments, the subject is an infant or less than 1 year of age. In other embodiments, the subject is an adult. In some embodiments, the subject is about 20 years of age, about 25 years of age, about 30 years of age, about 35 years of age, about 40 years of age, about 45 years of age, about 50 years of age, about 55 years of age, about 60 years of age, about 65 years of age, about 70 years of age, about 75 years of age, about 80 years of age, or more than 80 years of age. In some embodiments, the subject is immunocompromised or has an intact immune system (immunocompetent). Exemplary areas for administration include, without exclusion, the face, neck, antecubital fossa, and forearm.

[0060] In some embodiments, compositions described herein are applied to the skin, such as at cosmetic irregularities and around irregularity area, or at areas of unblemished skin (non irregularity areas) to prevent irregularities from forming. In some embodiments, compositions described herein are used to reduce irregularity size or extent. In some embodiments, compositions described herein are applied at one time (daily) or at multiple times throughout the day. In some embodiments, compositions are be applied 2 times, 3 times, 4 times, or 5 times per day. In some embodiments, compositions are be applied every other day, daily over a week, every other day over a week, every week, 2 times per week, 3 times per week, 4 times per week, 5 times per week, 6 times per week, or 7 times per week. In some embodiments, compositions described herein are compositions are formulated as a unit dose for administration.

[0061] In some embodiments, compositions and methods described herein are compositions used to improve a cosmetic irregularity. Improvement of the cosmetic irregularity may result in reduced fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, scaly patches, roughness, acne, scars, irregular pigmentation, sun spots, liver spots, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, blistering, peeling, sloughing, flaking, or pore size; improved skin tone, skin firmness, tactile smoothness, suppleness, glow, visual smoothness, or radiance. In addition, improvement of the cosmetic irregularity with a composition or method described herein may reduce S. aureus in the skin of a subject in need thereof. In some embodiments, compositions and methods described herein provide for enhanced barrier function of the skin as measured by trans-epidermal water loss. Administrations described herein, e.g., topical, oral, or rectal, may reduce reoccurrences, so that additional incidents of the cosmetic irregularities of the skin are reduced in number, intensity, or frequency. The administration may increase the time of remission, such as the length of time between incidents. In some embodiments, an additional incident of cosmetic irregularity does not occur for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks following application. In some embodiments, an additional incident of cosmetic irregularity does not occur for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months following the topical application. Kits

[0062] In some embodiments, compositions described herein are provided as a component of a kit. The purified viable bacteria can be provided in a growth medium, lyophilized form, or as frozen cells. Thus, the kit can include a container including a purified viable bacteria and, optionally, an additional metabolite for improvement of a skin condition described herein.

[0063] In some embodiments, the kit can include the components needed to produce a composition, such as one container including the bacteria and one container including a carrier for suspending the bacteria thereof. The carrier can be, for example, a buffered saline solution or a sucrose solution. In other embodiments, the kit can include a container including the bacteria, and a second container including a carrier, and a device, such as, but not limited to, a syringe, for measuring the carrier. In some embodiments, the kit includes a device, such as, but not limited to, a spray nozzle or a bandage, for topical application of the bacteria once it is suspended in the carrier.

[0064] Optionally, such a kit includes additional components including packaging, instructions and various other reagents, such as additional buffers or other therapeutic ingredients. The kit can include a container and a label or package insert on or associated with the container.

Suitable containers can include, for example, bottles, vials, tubes, etc. The containers can be formed from a variety of materials, such as glass or plastic. The container can hold a composition including bacteria effective for treating a skin described herein. In some embodiments, the container can have a sterile access port. For example, the container can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle. [0065] The label or package insert indicates that the composition can be used for treating the particular condition, such as a cosmetic irregularity. The label or package insert typically will further include instructions for use. The package insert typically includes instructions customarily included in commercial packages of cosmetic products that contain information about the indications, usage, dosage, administration, contraindications and/or warnings concerning the use of such cosmetic products. Non-limiting examples of instructions include information on the amount of the carrier to add to the vial containing the bacteria, instructions for suspending the bacteria in the carrier, and instructions for topical application to the skin. In some embodiments described herein, the application is sprayed on the skin, swabbed on the skin, or introduced in suspension onto a bandage for application to the skin.

Assessment

[0066] Change in cosmetic irregularities following administration of compositions as described herein can be assessed according to any assessment methodology as is known in the art. In some embodiments, change in cosmetic irregularities can be assessed according to SCORAD (SCORe Atopic Dermatitis) criteria. In some embodiments, change in cosmetic irregularities can be assessed according to EASI (Eczema Area and Severity Index) criteria. In some embodiments, change in cosmetic irregularities can be assessed using comparison of a condition before and after administration.

[0067] The SCORAD index is an assessment method for Atopic Dermatitis (AD). To measure the extent of AD, the rule of nines is applied on a front/back drawing of the patient’s inflammatory lesions. The intensity part of the SCORAD index consists of six items: erythema, oedema/papulation, excoriations, lichenification, oozing/crusts and dryness. Each item can be graded on a scale 0-3. The subjective items include daily pruritus and sleeplessness. Both subjective items can be graded on a 10-cm visual analogue scale. The maximum subjective score is 20. The SCORAD index formula is: A/5 + 7B/2 + C. In this formula A is defined as the extent (0-100), B is defined as the intensity (0-18) and C is defined as the subjective symptoms (0-20). The maximum SCORAD score is 103.

[0068] The Eczema Area and Severity Index (EASI) is a validated scoring system that grades the physical signs of atopic dermatitis/eczema. The Eczema Area and Severity Index (EASI) is a tool for the measurement of severity of atopic dermatitis. It ranges from 0 (no eczema) to 72 by grading the regional severity of signs of erythema, edema/papulation, excoriation, or lichenification on a scale of 0-3.

[0069] Condition of a subject skin before and after administration of a composition described herein can be compared to a standard skin model, for example, baby skin. Skin can be graded according to clinical, functional, or histologic criteria. Criteria can be objective or subjective. Objective criteria can comprise smoothness, firmness, even coloration, normal texture, and absence of any clinically evident disease. Subjective criteria can comprise proper hydration and normal tolerance. Grading of each element in the scoring system [minimal (1), average (2), maximal (3)], and subsequently the final score [excellent (12 to 15), average (7 to 11), poor (<7)] are done with reference to the healthy skin model defined

[0070] In some embodiments, provided herein are methods for obtaining a cosmetic skin assessment score for a subject. Provided herein are methods, wherein a skin assessment score is compared to a threshold. In cases wherein the score fails to meet the threshold, methods comprise providing instructions for administering a composition described herein, thereby providing a beneficial change in the cosmetic skin assessment score. In some embodiments, the cosmetic skin assessment score uses SCORAD criteria, EASI criteria, or comparison to a standard skin model. [0071] The following examples are set forth to illustrate more clearly the principle and practice of embodiments disclosed herein to those skilled in the art and are not to be construed as limiting the scope of any claimed embodiments. Unless otherwise stated, all parts and percentages are on a weight basis.

EXEMPLARY EMBODIMENTS

[0072] Embodiment 1. A composition for alteration of skin appearance, comprising: at least one species of gram-negative bacteria isolated from skin of a donor and present in an amount sufficient to alter skin appearance in a subject; and a carrier, wherein the composition is in a topical dosage form.

[0073] Embodiment 2. The composition of embodiment 1, wherein the at least one species of gram-negative bacteria is present in an amount sufficient to prevent or reduce fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, scaly patches, roughness, acne, scars, irregular pigmentation, sun spots, liver spots, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, tactile smoothness, suppleness, glow, visual smoothness, or radiance.

[0074] Embodiment 3. The composition of embodiment 1, wherein the carrier comprises sucrose.

[0075] Embodiment 4. A composition for moisturization of skin, wherein the composition comprises: at least one species of gram-negative bacteria isolated from skin of a donor; and a humectant or emollient, wherein the composition is in a topical dosage form.

[0076] Embodiment 5. The composition of embodiment 4, wherein the humectant is glycerin, propylene glycol, dipropylene glycol, polyethylene glycol, butylene glycol, hexane triol.

[0077] Embodiment 6. The composition of embodiment 4, wherein the emollient is urea, ammonium lactate, vitamin A, vitamin D, or vitamin E.

[0078] Embodiment 7. The composition of embodiment 4, further comprising the humectant and the emollient.

[0079] Embodiment 8. A composition for cleansing of skin, comprising: at least one species of gram-negative bacteria isolated from skin of a donor; and a detergent, wherein the composition is in a topical dosage form.

[0080] Embodiment 9. The composition of embodiment 8, wherein the detergent comprises sodium laurel sulfate, sodium tallowate, sodium cocoate, or sodium palmate.

[0081] Embodiment 10. A composition for skin protection, comprising: at least one species of gram-negative bacteria isolated from skin of a donor; and an agent to reduce sunlight exposure, wherein the composition is in a topical dosage form.

[0082] Embodiment 11. The composition of embodiment 10, wherein the agent to reduce sunlight exposure is octyl methoxyl cinnamate or butyl methoxy benzoylmethane.

[0083] Embodiment 12. The composition of any one of embodiments 1 to 11, further comprising an emulsifier.

[0084] Embodiment 13. The composition of embodiment 12, wherein the emulsifier is glycerol stearate, polyglyceryl-3 hydroxylauryl ether, polysorbate 60, or sorbitol polyethylene glycol. [0085] Embodiment 14. The composition of any one of embodiments 1 to 13, further comprising an antioxidant.

[0086] Embodiment 15. The composition of embodiment 14, wherein the antioxidant is vitamin C, vitamin E, vitamin A, or a tocopherol.

[0087] Embodiment 16. The composition of any one of embodiments 1 to 15, wherein the at least one species of gram-negative bacteria is present in an amount of up to about 10² CFUs. [0088] Embodiment 17. The composition of any one of embodiments 1 to 16, wherein the at least one species of gram-negative bacteria is present in an amount of up to about 1010 CFUs. [0089] Embodiment 18. The composition of any one of embodiments 1 to 17, wherein the at least one species of gram-negative bacteria comprises Roseomonas mucosa, Acinetobacter radioresistens, Moraxella osloensis, Pantoea septica, Pseudomonas luteola, Pseudomonas aeruginosa, or Pseudomonas oryzihabitans .

[0090] Embodiment 19. The composition of any one of embodiments 1 to 17, wherein the at least one species of gram-negative bacteria comprises a species of Roseomonas.

[0091] Embodiment 20. The composition of embodiment 19, wherein the species of Roseomonas is Roseomonas mucosa.

[0092] Embodiment 21. The composition of embodiment 19, wherein the species of Roseomonas is Roseomonas aerilata, Roseomonas aerophila, Roseomonas aestuarii, Roseomonas alkaliterrae, Roseomonas aquatic, Roseomonas cervicalis, Roseomonas fauriae, Roseomonas frigidaquae, Roseomonas gilardii, Roseomonas lacus, Roseomonas ludipueritiae, Roseomonas mucosa, Roseomonas pecuniae, Roseomonas rhizosphaerae, Roseomonas riguiloci, Roseomonas rosea, Roseomonas soli, Roseomonas stagni, Roseomonas terrae, or Roseomonas vinacea.

[0093] Embodiment 22. The composition of any one of embodiments 1 to 17, wherein the at least one species of gram-negative bacteria are viable. [0094] Embodiment 23. The composition of any one of embodiments 1 to 17, wherein the at least one species of gram-negative bacteria are purified.

[0095] Embodiment 24. The composition of any one of embodiments 1 to 17, wherein the at least one species of gram-negative bacteria are isolated.

[0096] Embodiment 25. The composition of any one of embodiments 1 to 24, wherein the topical dosage form is a solid, semisolid, cream, gel, foam, ointment, or liquid.

[0097] Embodiment 26. A method of preventing or reducing a skin condition in a subject, comprising topically administering to a subject the composition of any one of embodiments 1 to 25.

[0098] Embodiment 27. The method of embodiment 26, wherein the skin condition is fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, scaly patches, roughness, acne, scars, irregular pigmentation, sun spots, liver spots, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, tactile smoothness, suppleness, glow, visual smoothness, or radiance.

[0099] Embodiment 28. The method of embodiment 26, wherein the subject is an infant, child, or adult.

[0100] Embodiment 29. The method of embodiment 26, wherein the composition is administered to the subject at least two times per a week.

[0101] Embodiment 30. The method of embodiment 26, wherein the composition is administered to the subject every other day over a week.

[0102] Embodiment 31. The method of embodiment 26, wherein the composition is administered to the subject daily.

[0103] Embodiment 32. The method of embodiment 26, wherein the composition is administered to the subject multiple times a day.

EXAMPLES

Example 1: Gram-negative Bacterial Collection and Identification [0104] Two FloqSwabs (Copan, Brescia, Italy) moistened in sterile phosphate buffered saline (PBS; Corning Cellgro, Coming, NY) are rubbed on the subject’s skin (at the antecubital fossa) vigorously for 15-30 seconds. One swab is placed into a 15mL conical tube (Corning Life, Corning, NY) with 2mL of sterile Hank’s balanced salt solution (HBSS; Sigma- Aldrich) containing vancomycin (300ug/mL) and amphotericin B (5ug/mL; Sigma-Aldrich, St. Louis, MO) to inhibit growth of gram-positive bacteria and fungi. The remaining swab is placed into a 15mL conical tube containing 2mL of R2A (Reasoner's 2A) broth (Teknova, Hollister, CA) with similar concentrations of vancomycin and amphotericin B. The tubes, with swabs left in place, are then incubated at 32°C with constant shaking for 48-72 hours before plating lOOuL from each tube onto an R2A agar plate (Remel, Lenexa, KS). Colonies are then taken for species identification by mass spectrometry using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis. Bacterial protein extraction for MALDI-TOF MS using the BioTyper (v3.1, Bruker Daltonics Inc., Billerica, MA) is performed by the NIH Clinical Center microbiology lab using previously described methods (Lau et ak, Journal of clinical microbiology 52, 2804-2812 (2014)), instrument settings and calibration (Lau et al., Journal of clinical microbiology 51, 828-834 (2013); Youn et al ., Journal of clinical microbiology , (2015)). BioTyper identification is supplemented by additional mass spectra profiles provided by several NIH developed databases (Lau et al ., 2014, supra; Stevenson et al., Journal of clinical microbiology 48, 3482-3486 (2010); Myles et al., Nature immunology 14, 804-811 (2013)). All R. mucosa isolates used for subsequent assays are verified by MALDI-TOF analysis.

Example 2: Generation and characterization of strains of Roseomonas mucosa [0105] Provided herein are an A/G allelic discrimination assay and a T/C allelic discrimination assay. Briefly, the assay provides for selection 3 Roseomonas mucosa isolates. Samples of R mucosa isolates are collected, cultured, and prepared for topical administration. Additional details regarding preparation can be found in published PCT application, WO 2020/023072 . [0106] Genomes from the three isolates of R mucosa have regions of sequence specific to each of the three isolates, as shown in Table 3 (bases specific to each strain are in bold and underlined).

Table 3.

[0107] Primers are designed to amplify the region where strain specific variation is identified. A Custom TaqMan® SNP Genotyping Assays, Non-human, SM kit and protocol are used to perform an analysis for detection of each strain. Briefly, DNA from each isolate is subjected to PCR where the primers are SEQ ID NO: 4 (CACCGGACAGCAGGCT), and SEQ ID NO: 5 (GCGGTGGCTTAGCATCATC). Amplification products are subjected to an allelic discrimination assay. In a first comparison, the following reporters are used: SEQ ID NO: 6 (CACCCCATCCTCG) and SEQ ID NO: 7 (CACCCCGTCCTCG). This is an A/G allelic discrimination assay. In a second comparison, the following reporters were used: SEQ ID NO: 8 (CCCTCCACCCCATCCT) and SEQ ID NO: 9 (CCCTCCACTCCATCCT). This is a T/C allelic discrimination assay.

Example 3: Assessment of Clinical Improvement in a Skin Condition [0108] Twenty subjects with atopic dermatitis (AD) were administered a regimen of Roseomonas mucosa. Fifteen of the subjects received a dose escalation regimen of 10³ colony forming units (CFU) twice weekly per body site for 4 weeks, followed by 10⁴ CFU twice weekly for four weeks. Five subjects received a dose of 10⁵ CFU per body site of R. mucosa twice weekly for 8 weeks. (See, FIG. 2A) R mucosa in sucrose was applied directly to affected areas. Subjects were assessed for improvements in severity using standard measurement criteria. Both treatment groups showed improvement according to SCORAD (FIG. 2B) and EASI (FIG. 2C) criteria. Under both assessment criteria, subjects showed about 30-40% improvement at the lower doses of 10³-10⁴ CFU in the dose escalation group. For additional information, see Myles et al, Sci. Transl. Med. 12 (2020).

Example 4: Skin Condition Assessment Using the Compositions Described Thereof [0109] Volunteers are selected to examine the efficacy and tolerability of the bacterial compositions in subjects with various skin conditions. The skin conditions comprise fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

[0110] Subjects are instructed to apply the composition on their facial skin twice daily for twelve weeks. Clinical evaluations are conducted at baseline (Visit 1), week 4 (Visit 2), week 8 (Visit 3) and week 12 (Visit 4). The following procedures are conducted at each visit.

[0111] Efficacy Assessments: skin conditions are clinically graded using a ten-point scale, on each subject's right and left facial area (where 0=none, 0.5-3.5=mild, 4-6.5=moderate and 7- 9=severe). All grading assessments are performed by the same investigator at each visit to ensure grading consistency.

[0112] Safety Assessments: Tolerability of the compositions are assessed at all visits by the reporting of adverse events and by objective and subjective assessments. Objective irritation (erythema, edema and scaling) is assessed by the investigator whereas subjective irritation (buming/stinging, itching and tingling) is assessed by the subject, all using a 4-point scale (where 0=none, l=mild, 2=moderate and 3=severe).

[0113] Standardized Photography: Standardized, digital images are taken of the subject's left, right and frontal facial views using raking light (standard color).

[0114] Self-Assessment Questionnaires: Subjects complete a Self-Assessment Questionnaire at Week 12, where they rate their facial skin condition and the compositions’ efficacy on a four- point scale (Strongly Agree, Agree, Disagree, Strongly Disagree). Subjects rate their overall satisfaction with the compositions on a four-point scale where l=Excellent, 2=Good, 3=Fair and 4=Poor.

[0115] Statistical Analysis: Mean clinical grading scores at weeks 4, 8 and 12 are compared to mean baseline scores using a student's paired t-test. Changes from baseline are considered significant at the p<0.05 level. For the subject Self-Assessment Questionnaires, the percent incidence of positive responders is reported for the treatment serum's efficacy and aesthetic attributes sections.

Example 5: Administration of Bacterial Compositions for Skin Condition Assessment [0116] Bacterial compositions comprising 10² CFU bacteria in combinations described in Table 4 are separately administered in a study as described in Example 4. Assessments of skin condition before and after treatment are made as described in Example 4.

Table 4.

[0117] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. A composition for moisturization of skin, wherein the composition comprises: at least one species of gram-negative bacteria, wherein the at least one species of gram negative bacteria is purified and viable; and a humectant or emollient, wherein the composition is in a topical dosage form.

2. The composition of claim 1, wherein the at least one species of gram-negative bacteria is isolated from skin of a donor.

3. The composition of claim 1, wherein the at least one species of gram-negative bacteria is a commensal organism.

4. The composition of claim 1, wherein the humectant is glycerin, propylene glycol, dipropylene glycol, polyethylene glycol, butylene glycol, hexane triol.

5. The composition of claim 1, wherein the emollient is urea, ammonium lactate, vitamin A, vitamin D, or vitamin E.

6. The composition of claim 1, further comprising the humectant and the emollient.

7. A composition for cleansing of skin, comprising: at least one species of gram-negative bacteria, wherein the at least one species of gram negative bacteria is purified and viable; and a detergent, wherein the composition is in a topical dosage form.

8. The composition of claim 7, wherein the at least one species of gram-negative bacteria is isolated from skin of a donor.

9. The composition of claim 7, wherein the at least one species of gram-negative bacteria is a commensal organism.

10. The composition of claim 7, wherein the detergent comprises sodium laurel sulfate, sodium tallowate, sodium cocoate, or sodium palmate.

11. A composition for reduction in sunlight exposure or damage from sunlight exposure, comprising: at least one species of gram-negative bacteria, wherein the at least one species of gram negative bacteria is purified and viable; and an agent to reduce sunlight exposure or damage from sunlight exposure, wherein the composition is in a topical dosage form.

12. The composition of claim 11, wherein the at least one species of gram-negative bacteria is isolated from skin of a donor.

13. The composition of claim 11, wherein the at least one species of gram-negative bacteria is a commensal organism.

14. The composition of claim 11, wherein the agent to reduce sunlight exposure is octyl methoxyl cinnamate or butyl methoxy benzoylmethane.

15. The composition of any one of claims 1 to 14, further comprising an emulsifier.

16. The composition of claim 15, wherein the emulsifier is glycerol stearate, polyglyceryl-3 hydroxylauryl ether, polysorbate 60, or sorbitol polyethylene glycol.

17. The composition of any one of claims 1 to 16, further comprising an antioxidant.

18. The composition of claim 17, wherein the antioxidant is vitamin C, vitamin E, vitamin A, or a tocopherol.

19. The composition of any one of claims 1 to 18, wherein the at least one species of gram negative bacteria is present in an amount of up to about 10² CFUs.

20. The composition of any one of claims 1 to 19, wherein the at least one species of gram negative bacteria comprises Roseomonas mucosa , Acinetobacter radioresistens , Moraxella osloensi , Pantoea septica , Pseudomonas luteola , Pseudomonas aeruginosa , or Pseudomonas oryzihabitans .

21. The composition of any one of claims 1 to 19, wherein the at least one species of gram negative bacteria comprises a species of Roseomonas.

22. The composition of claim 21, wherein the species of Roseomonas is Roseomonas mucosa.

23. The composition of any one of claims 1 to 19, wherein the at least one species of gram negative bacteria are isolated.

24. The composition of any one of claims 1 to 23, wherein the topical dosage form is a solid, semisolid, cream, lotion, gel, foam, ointment, powder, or liquid.

25. The composition of claim 24, wherein the topical dosage form comprises a water-based formulation.

26. A method of preventing or reducing a skin condition in a subject, comprising topically administering to a subject the composition of any one of claims 1 to 25.

27. The method of claim 26, wherein the skin condition comprises fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

28. The method of claim 26, wherein the subject is an infant, child, or adult.

29. The method of claim 26, wherein the composition is administered to the subject at least two times per a week.

30. The method of claim 26, wherein the composition is administered to the subject every other day over a week.

31. The method of claim 26, wherein the composition is administered to the subject daily.

32. The method of claim 26, wherein the composition is administered to the subject multiple times a day.

33. The method of claim 26, wherein the composition is applied to a bandage for topical administration to the subject.

34. A method of preventing or reducing a skin condition in a subject, comprising topically administering at least one strain of Roseomonas mucosa to a subject, wherein the Roseomonas mucosa is purified and viable , and wherein the skin condition comprises: fine line occurrence, fine line depth, wrinkle occurrence, wrinkle depth, nasolabial folds, scaly patches, roughness, acne, scars, redness, irregular pigmentation, decreased tightness, decreased skin barrier, age spots, sun spots, liver spots, photo damage, sunlight exposure, damage from sunlight exposure, environmental damage, solar lentigines, melasma, poikiloderma, actinic keratoses, lentigo maligna, periorbital hyperpigmentation, shine, sheen, oily appearance, decrease of skin’s ability to retain moisture, abnormal skin epidermal thickness, reduction of dermal epidermal junction, blistering, peeling, sloughing, flaking, pore size, skin tone, skin firmness, skin texture, skin elasticity, tactile smoothness, suppleness, glow, visual smoothness, radiance or a combination thereof.

35. The method of claim 34, wherein the subject is an infant, child, or adult.

36. The method of claim 34, wherein the composition is administered to the subject at least two times per a week.

37. The method of claim 34, wherein the composition is administered to the subject every other day over a week.

38. The method of claim 34, wherein the composition is administered to the subject daily.

39. The method of claim 34, wherein the composition is administered to the subject multiple times a day.

40. The method of claim 34, wherein the composition is administered to the subject 2, 3, 4, 5, 6 or more times a day.

41. The method of claim 34, wherein the Roseomonas mucosa is present in an amount of up to about 10² CFUS.

42. The method of claim 34, wherein the Roseomonas mucosa is in a topical dosage form.

43. The method of claim 42, wherein the topical dosage form is a solid, semisolid, cream, lotion, gel, foam, ointment, powder, or liquid.

44. The method of claim 34, wherein the administering provides for a beneficial shift in a cosmetic skin assessment score.

45. The method of claim 44, wherein the cosmetic skin assessment score is measured using SCORAD criteria, EASI criteria, or comparison to a standard skin model.