Disposable personal care products comprising bacteriophage-coated nonwoven fabrics for preventing and reducing skin problems caused by bacteria

Abstract

Provided herein are compositions of phage combinations bound to surface-active polymer substrates. The polymer substrates can be incorporated into personal care products. Also provided herein are methods for binding the phage combinations to surface-active polymer substrates. Methods for selecting the phage combinations are also described. Also provided are methods for using the phage combination compositions to treat one or more skin conditions.

Description

[Technical Field]

[0001] 1. Cross-reference to related applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 385,398, filed November 29, 2022, which is incorporated herein by reference in its entirety.

[0002] 2. Sequence Listing This application contains a Sequence Listing that has been submitted electronically in XML format and is incorporated herein by reference in its entirety. Said XML copy, created on November 28, 2023, is named 54391WO_CRF_sequencelisting and is 1,895,212 bytes in size. [Background technology]

[0003] 3.Background Common skin problems such as rashes, sensitive skin, itching, and discoloration are often associated with the use of disposable personal care products made from occlusive nonwoven fabrics. These problems are usually associated with contact dermatitis and manifest as common consumer complaints such as diaper rash, maskne, sanitary napkin rash, and incontinence-related rashes. The personal care products associated with these complaints can adhere to the skin for hours at a time, elevating skin pH, temperature, and moisture. The design of the personal care products themselves and the structure of the nonwoven fabrics worn closest to the skin have been optimized to wick moisture away from the skin, absorb excess fluid distal to the skin, and reduce irritation from friction. Nevertheless, these products induce changes in skin physiology and the skin microbiome, leading to contact dermatitis and related problems. Skin occlusion increases the number of viable bacteria on the skin. It has also been found to distort the composition of the bacterial communities growing on the skin. Only recently has high-throughput DNA sequencing enabled accurate and quantitative measurement of the skin microbiome.

[0004] High-throughput DNA sequencing methods, such as shotgun metagenomics, allow for the accurate identification of microbial strains and their abundance on the skin. In recent years, these sequencing methods have become automated and their costs have fallen sufficiently to allow for the sequencing of skin microbiomes across large consumer populations. Quantitative and accurate population-level data derived from testing consumer skin microbiomes can reveal which microorganisms are associated with specific types of skin problems, such as contact dermatitis. Common harmful microbial species can also be targeted for elimination. Thus, large-scale skin microbiome data allows for the precise engineering of skin microbiomes across large consumer populations.

[0005] Antimicrobial agents have been added to consumer products for decades, but some have recently been banned by the FDA for certain consumer applications. Triclosan and triclocarban were particularly common broad-spectrum antimicrobials added to consumer products such as plastic toys, cutting boards, and hygiene products like antibacterial hand soaps. Some of these products included disposable personal care products made from nonwoven fabrics, such as sanitary napkins. However, antimicrobial agents have been found to be absorbed through the skin, persist in the environment, and exacerbate the spread of antibiotic resistance among bacteria found in society. In 2016, the FDA banned triclosan and triclocarban from over-the-counter consumer antibacterial hand washes and body washes. Broad-spectrum small-molecule antibacterial agents have lost favor in the consumer market, largely due to their negative side effects on the skin and gut microbiome, the human endocrine system, and antibiotic stewardship. Clear consumer demand exists for effective antibacterial agents in consumer products, including disposable consumer products made from nonwoven fabrics, but acceptable options are limited.

[0006] New precision antimicrobial agents with minimal side effects are needed to address contact dermatitis and other conditions caused by disposable personal care products constructed from nonwoven fabrics. Summary of the Invention

[0007] 4. Overview Personal products, such as disposable personal care products composed of nonwoven or nonwoven materials, such as sanitary napkins, diapers, and face masks, occlude the skin, increase total bacterial counts, promote contact dermatitis, and cause consumer complaints such as diaper rash, maskne, sanitary napkin rash, and incontinence-related rash. Broad-spectrum, small-molecule antimicrobial agents previously used to reduce the problem have either been banned by the FDA in some instances of consumer use or have fallen out of favor with consumers.

[0008] In one aspect, a composition is disclosed comprising a phage combination bound to a polymer fabric, wherein the phage combination comprises one or more sets of phages that infect one or more bacterial species, the phages being selected from Myoviridae, Podoviridae, and Siphoviridae, and the polymer fabric being selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

[0009] A composition comprising a phage combination bound to a polymer fabric, wherein the phage combination comprises one or more sets of Caudoviricetes phages, and the polymer fabric is a polyethylene terephthalate (PET) nonwoven fabric.

[0010] In one aspect, a composition is disclosed comprising a phage combination bound to a polymer fabric, wherein the phage combination comprises phages that infect at least Staphylococcus aureus, and the polymer fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

[0011] In another aspect, a composition is disclosed comprising a phage combination bound to a polymer fabric, wherein the phage combination comprises at least phages that infect Escherichia coli, and the polymer fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

[0012] In another aspect, a composition is disclosed comprising a phage combination bound to a polymer fabric, wherein the phage combination comprises one or more sets of Caudoviricetes phages, and the polymer fabric is selected from the group consisting of polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, polycaprolactone (PCL) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

[0013] In one aspect, a composition is disclosed comprising a phage combination bound to a polymer fabric, the phage combination comprising one or more sets of phages that infect one or more bacterial species.

[0014] According to any one of the embodiments, the phage is of the genus Pahexavirus or Rosenblumvirus. The phage may be a tailed virus. In some embodiments, the phage is at least one of Myoviridae or Podoviridae. In some embodiments, the phage is a T4 myovirus. In some embodiments, the phage is a podovirus.

[0015] According to any one of the embodiments, the set or sets of phages infect at least one common target bacterial species.

[0016] According to any one of the embodiments, each set of phages comprises at least two, three, four, five or more phages selected from one or more phage libraries, wherein the one or more phage libraries comprise phages from one or more genera.

[0017] According to any one of the embodiments, the phage combination is non-covalently or covalently bound to a polymer fabric, which may be a synthetic nonwoven fabric.

[0018] According to any one of the embodiments, the phage combination is deposited on at least one material that forms at least a part of a personal care product. In some embodiments, the personal care product is selected from the group consisting of a face mask, a sanitary napkin, a tampon, a baby diaper, an adult incontinence product, or a cosmetic facial pad or wipe, a fingernail wipe, or a patient bath hygiene fabric.

[0019] According to any one of the embodiments, the bacterial species is generally associated with contact dermatitis. In some embodiments, the bacterial species is selected from a skin microbiota biobank. In some embodiments, the bacterial species is derived from the skin microbiota isolated from the subject's biological sample.

[0020] According to any one of the embodiments, the one or more bacterial species are selected from a group of microorganisms including Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, Corynebacterium tuberculostearicum, or a combination thereof.

[0021] According to any one of the embodiments, the phage is a phage that infects Escherichia coli.

[0022] According to any one of the embodiments, the phage is a phage that infects Staphylococcus aureus.

[0023] According to any one of the embodiments, the polymer fabric further comprises a fabric selected from the group consisting of polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, polycaprolactone (PCL) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

[0024] In one aspect, a method of selecting a phage combination according to any one of the embodiments is disclosed.

[0025] In one aspect, a method for binding a phage combination according to any one of the embodiments to a polymer nonwoven fabric is disclosed. In some embodiments, the binding of the phage combination is achieved by impregnating the phage capsid onto the surface of the polymer nonwoven fabric. The phage capsid can be absorbed onto the surface of the polymer fabric by electrostatic charging. In some embodiments, the tail is exposed away from the fabric.

[0026] In some embodiments, binding of phage combinations is achieved by misting, chemical binding, physical adsorption, bulk mixing, plasma-treated surface binding, or a combination thereof.

[0027] In some embodiments, binding of the phage combination is achieved by plasma-treated surface binding.

[0028] In one aspect, a method of preparing a personal care product is disclosed, comprising contacting a polymeric nonwoven fabric with a composition comprising a phage combination, wherein the phage combination comprises one or more sets of phages of Caudoviricetes phages that infect one or more bacterial species, and wherein the polymeric nonwoven fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

[0029] In one aspect, a method of preparing a personal care product is disclosed, the method comprising: (a) contacting a polymeric nonwoven fabric with a plasma composition to produce a plasma-treated nonwoven fabric, wherein the polymeric nonwoven fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric; (b) immobilizing a phage combination comprising at least one or more sets of Caudoviricetes phages on the polymeric nonwoven fabric, wherein the phage combination comprises one or more sets of phages that infect one or more bacterial species; and (c) incorporating the polymeric nonwoven fabric into a personal care product.

[0030] In one aspect, a polymeric nonwoven fabric is disclosed comprising a phage combination bound to the polymeric fabric, wherein the phage combination comprises one or more sets of Caudoviricetes phages that infect one or more bacterial species, and the polymeric fabric is selected from polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

[0031] In one aspect, a method for manufacturing a personal care product for reducing contact dermatitis, reducing skin infections, and / or reducing body odor is disclosed, wherein the personal care product comprises a polymeric nonwoven fabric, the polymeric nonwoven fabric comprises a phage combination bound to the nonwoven fabric selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric, the phage combination comprising one or more sets of Caudoviricetes phages that infect one or more bacterial species. In some embodiments, the personal care product is selected from the group consisting of a face mask, a sanitary napkin, a tampon, a baby diaper, an adult incontinence product, or a cosmetic facial pad or wipe, a fingernail wipe, or a patient bathing hygiene textile.

[0032] According to any one of the embodiments, the phage of the method, or the phage of the polymer nonwoven fabric, or the phage of the personal care product comprises a nucleic acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any one of SEQ ID NOs: 1-132.

[0033] According to any one of the embodiments, the phage of the method, or the phage of the polymer nonwoven fabric, or the phage of the personal care product comprises a nucleic acid sequence having at least 95% identity to any one of SEQ ID NOs: 1-132.

[0034] According to any one of the embodiments, the phage of the method, or the phage of the polymer nonwoven fabric, or the phage of the personal care product comprises a nucleic acid sequence having the sequence of any one of SEQ ID NOs: 1-132.

[0035] Described herein are compositions of bacteriophage mixtures and methods of using the bacteriophages (phages) as antimicrobial agents for consumer products (such as disposable consumer products composed of one or more nonwoven fabrics).

[0036] Thus, in one aspect, described herein are compositions of one or more phages that infect bacteria on the surface of the skin associated with contact dermatitis. In one aspect, described herein are methods for selecting one or more phage combinations to reduce the concentration of bacteria commonly associated with contact dermatitis. In one aspect, described herein are methods for attaching one or more phage combinations to nonwoven fabrics incorporated into disposable personal care products.

[0037] These phages infect bacterial species, including members of Gram-positive and Gram-negative bacteria. In certain embodiments, the phages infect one or more species, including, but not limited to, members of the phyla Bacillotia, Proteobacteria, and Actinobacteria. In certain embodiments, the phages infect one or more species, including, but not limited to, members of the families Staphylococcus, Streptococcaceae, Enterobacteriaceae, Enterococcaceae, Propionibacteriaceae, and Corynebacteriaceae. In some embodiments, the phages infect one or more species, including, but not limited to, members of the genera Staphylococcus, Streptococcus, Escherichia, Enterococcus, Cutibacterium, and Corynebacterium. For example, the phage may infect bacterial species such as Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, and / or Corynebacterium tuberculostearicum. In certain embodiments, the phage infects one or more fungi or viruses (including dsDNA viruses). In some embodiments, the phage infects Malassezia yeast. In certain embodiments, the phage infects mites (e.g., Demodex mites). In some embodiments, the phage infects a population of microorganisms including bacteria, fungi, viruses, and / or mites.

[0038] In the compositions or methods of the above embodiments, the phage is non-covalently bound to a surface-active nonwoven fabric incorporated into a personal care product, although it may also be covalently bound. In some embodiments, the personal care product is disposable, non-disposable, or biodegradable. The phage is bound such that the capsid adsorbs to the surface of the nonwoven fabric due to the electrostatic charge, leaving the tail exposed away from the surface of the fabric. When the nonwoven fabric of the personal care product comes into contact with skin, the exposed phage tail can initiate infection of any susceptible bacteria on the skin.

[0039] In any of the preceding embodiments, the phage combination is attached to a plasma surface-treated synthetic polymer nonwoven, including polylactic acid (PLA), polyester (PE), acrylic, olefin, polyhydroxyalkanoate (PHA), polyethylene terephthalate (PET), polypropylene (PP), and polypropylene carbonate (PPC). The surface-treated nonwoven is incorporated into a disposable skin-contacting personal care product, such as a face mask, sanitary napkin, tampon, baby diaper, or adult incontinence product.

[0040] In the composition or method of any preceding embodiment, the phage combination is bound to a natural polymer substrate, non-limiting examples of which are silk, wool, DNA, cellulose, and protein.

[0041] In any of the preceding embodiments, target bacterial strains from the species Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, and / or Corynebacterium tuberculostearicum are identified from a large-scale shotgun metagenomic dataset of skin microbiome samples collected from patients with contact dermatitis. In some embodiments, phages infecting these strains are isolated from environmental samples. In some embodiments, phage isolates are amplified on the target bacterial strain. DNA extracts from the isolates are sequenced to screen the phage genomes for genes associated with antibiotic resistance, virulence, lysogeny, or transduction. The screened phages are tested for host range and combined to maximize host range. In certain embodiments, sets of phages infecting the same host species are tested in pairs to prevent bacterial regrowth in broth culture. In certain embodiments, pairs are selected that prevent bacterial regrowth, hi some embodiments, more than two phage or two pairs of phage are selected that prevent bacterial regrowth.

[0042] In any preceding embodiment of the composition or method, the phage are concentrated in a low-salt phage buffer prior to binding to the nonwoven fabric. In some embodiments, a phage surface treatment is applied to the nonwoven fabric. In some embodiments, the phage buffer is misted onto the surface-treated nonwoven fabric and allowed to dry at room temperature. In some embodiments, the nonwoven fabric is incorporated into a disposable personal care product.

[0043] 5. A brief description of some figures in the drawing These and other features, aspects, and advantages of the present invention will become better understood with regard to the following description and accompanying drawings. [Brief explanation of the drawings]

[0044] [Figure 1] Figure 1 shows a phage bound to a surface-activated polymer fabric via its negatively charged capsid, with the tail free for binding to a phage receptor displayed on a target host bacterial species living in or on the subject's skin surface. Legend: (1) Skin surface; (2) Bacterial species targeted for removal; (3) Phage receptor on the target host bacterial species, including, but not limited to, Staphylococcus aureus, Escherichia coli, or Cutibacterium acnes; (4) Non-targeted commensal skin microorganisms; (5) Phage; (6) Negative dipole moment of the bacteriophage capsid; (7) Positive charge of the activated surface; (8) Nonwoven-activated surface functionalized with immobilized phage; (9) Podovirus-tailed phage, a phage morphotype distinct from myovirus-tailed phage.

[0045] [Figure 2] 2A-2C show that phages can bind to plasma-treated PET nonwoven fabric and inhibit bacterial growth. Phage inhibition of Escherichia coli strain 11303 growth on an agar plate is shown.

[0046] [Figure 3] 3A-3C show that phages can bind to plasma-treated PET nonwoven fabric and inhibit bacterial growth. Phage inhibition of Escherichia coli strain 11303 growth on an agar plate is shown.

[0047] [Figure 4] 4A-4C show that phages can bind to plasma-treated PET nonwoven fabric and inhibit bacterial growth. Phage inhibition of Staphylococcus aureus strain 100311 growth on an agar plate is shown.

[0048] [Figure 5]5A-5C show that phages can bind to plasma-treated PHB nonwoven fabric and inhibit bacterial growth. Phage inhibition of Staphylococcus aureus strain 100311 growth on an agar plate is shown.

[0049] [Figure 6] FIG. 6 shows the effect of T4 myoviridae bound to plasma-treated PET and PHB nonwovens on Escherichia coli strain 11303.

[0050] [Figure 7] FIG. 7 shows the effect of S. aureus podovirus Rosenblumvirus P00474 phage bound to plasma-treated PET and PHB nonwovens against Staphylococcus aureus strain 100311.

[0051] [Figure 8] FIG. 8 illustrates the feasibility of in-line phage treatment of nonwoven fabrics during production. DETAILED DESCRIPTION OF THE INVENTION

[0052] 6. Detailed Description 6.1. Bacteriophages Phages are viruses that infect bacteria. As used herein, the terms "phage" and "bacteriophage" are used interchangeably. Phages were discovered independently in 1915 and 1917 and have been used early in bacteriophage therapy to treat acute bacterial infections. Phages are ubiquitous in the environment and are important components of the skin microbiome. While they are effective antibacterial agents under certain conditions, they are precise in the species and strains of bacteria they kill. In part, broad-spectrum small-molecule antibiotics have been preferred over phages because there has been no data for the targeted use of phages. Only broad-spectrum antibacterial agents have been practical. Precision, quantitative, population-scale skin microbiome data has recast phages as candidates for precision antibacterial agents.

[0053] In various embodiments, the phage described throughout are of the class Caudoviricetes. In some embodiments, the phage belongs to the genus Pahexavirus. In some embodiments, the phage belongs to the genus Rosenblumvirus. In some embodiments, the phage is selected from Myoviridae, Siphoviridae, Podovirdae, or Rountreeviridae. In some embodiments, the phage is Myoviridae with a long, contractile tail. In some embodiments, the phage is Podoviridae with a short, non-contractile tail. In some embodiments, the phage is Siphoviridae with a long, non-contractile tail. In some embodiments, the phage has a nucleic acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any one of SEQ ID NOs: 1-132. In some embodiments, the phage has a nucleic acid sequence having at least 95% identity to any one of SEQ ID NOs: 1-132. In some embodiments, the phage has the nucleic acid of any one of SEQ ID NOs: 1-132.

[0054] In various embodiments, the phage infects a population of microorganisms, including bacteria, fungi, viruses, and / or mites. In some embodiments, the population of microorganisms is commonly found living on or in a skin sample obtained from a subject. In some embodiments, the population of microorganisms includes one or more microorganisms associated with acne. In some embodiments, the population of microorganisms includes one or more microorganisms associated with contact dermatitis, diaper rash, maskne, sanitary napkin rash, incontinence-related rash, inflammation, redness, eczema, rosacea, enlarged hair follicle pore size, rough skin, increased transepidermal water loss, skin discoloration, or an imbalance in the elasticity of the stratum corneum and the underlying dermis. In some embodiments, the population of microorganisms includes one or more microorganisms associated with aging skin. In some embodiments, the population of microorganisms includes Cutibacterium acnes bacteria. In some embodiments, the population of microorganisms includes Staphylococcus aureus bacteria. In some embodiments, the population of microorganisms includes Corynebacterium bacteria. In some embodiments, the population of microorganisms comprises Malassezia yeast. In some embodiments, the population of microorganisms comprises Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, and / or Corynebacterium tuberculostearicum.

[0055] Phages can be attached to nonwoven fabrics by activating the surface of the fabric using corona or plasma treatment. Surface activation imparts a charge to the nonwoven fabric, allowing phages to be noncovalently attached to it by electrostatic attraction. Phages consist of a capsid, which carries the genetic material necessary for infection, and a tail, which interacts with the bacterial host to facilitate infection. The phage capsid is electrically charged, and can be attached to the nonwoven fabric so that the phage tail is exposed and facing away from the surface of the fabric, facilitating infection. The nonwoven fabric is incorporated into a disposable personal care product where the product comes into contact with the skin. In this orientation, the phage tail can infect bacteria on the skin when the nonwoven fabric on the surface of the disposable consumer product comes into contact with the skin.

[0056] Figure 1 illustrates the use of a composition comprising a phage described herein for use in a personal care product. A subject's skin harbors various bacterial species living commensally in or on the skin surface (1). The phage targets specific types of bacteria (2) by recognizing phage receptors (3) displayed on the target host bacterial species. Note that non-targeted commensal skin microorganisms (4) remain undisrupted. The phage (5) displays a negatively charged capsid (6), which can bind to a positively charged (7) activated polymer fabric surface (8). The polymer fabric is a nonwoven fiber. The phage (5) is a myovirus-tailed phage. The phage (9) is a podovirus-tailed phage, a different phage morphotype from the myovirus-tailed phage. Upon phage binding, a polymer fabric (e.g., a surface-activated nonwoven fabric) (8) is functionalized with immobilized phage (5) or (9). This particular method of phage binding or deposition allows the phage tail to be displayed externally on the fabric, as the charged capsid binds to the activated polymer fabric surface, making the phage more effective in bacterial infection.

[0057] In some embodiments, the composition includes a bacteriophage for a fungal virus formulation comprising a set of nucleic acid sequence read data extracted from the genomes of multiple (e.g., 1, 2, 3, 4, 5, or more) bacteriophages or fungal viruses, wherein at least one virus is capable of lysing microorganisms of a population of microorganisms identified as living in or on the skin of a subject. Methods and compositions for identifying such microorganisms and bacteriophages that target them are described in WO2022198091A1, the entire contents of which are incorporated herein by reference.

[0058] 6.2. Composition Compositions comprising phages bound to fabrics are described herein. In certain embodiments, the phages are bound to fabrics, such as polymer fabrics, and incorporated into personal care products. The polymer fabrics may be surface-active nonwoven fabrics. The surface-active nonwoven fabrics may be plasma-treated. The fabrics may be synthetic, artificial, or natural.

[0059] In some embodiments, the polymer fabric comprises a degradable or non-degradable polymer. In some embodiments, the polymer fabric comprises polylactic acid, polyhydroxyalkanoate, or polycaprolactone. In some embodiments, the polymer fabric is a nonwoven fabric.

[0060] In some embodiments, the composition comprises a phage non-covalently bound to a surface-active nonwoven fabric incorporated into a disposable personal care product. In some embodiments, the composition comprises one or more sets of phage or fungal viral DNA sequences isolated from a subject's skin sample and, if necessary, incorporated onto the surface of the personal care product. In some embodiments, the composition comprises one or more sets of phage DNA isolated from a first bacteriophage and a second bacteriophage. In some embodiments, the first bacteriophage and the second bacteriophage are capable of lysing microorganisms in a population of microorganisms. In some embodiments, the first bacteriophage and the second bacteriophage individually or collectively prevent resistance or regrowth of microorganisms in vitro.

[0061] In some embodiments, polymer surfaces are modified to facilitate phage binding using plasma treatment alone, plasma treatment followed by activation with 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), plasma-initiated acrylic acid grafting, or plasma-initiated acrylic acid grafting with activation with EDC and sulfo-NHS.

[0062] In some embodiments, the first bacteriophage and the second bacteriophage are capable of killing, preventing resistance to, or regrowth of a population of microorganisms including, but not limited to, Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, and / or Corynebacterium tuberculostearicum.

[0063] In some embodiments, the first bacteriophage and the second bacteriophage have different host ranges. The different host ranges may include at least two C. acnes strains, or the different host ranges may include at least one C. acnes strain and at least one C. namnatense strain. In some embodiments, the different host ranges do not include C. granulosum strains. The first bacteriophage and the second bacteriophage may be lytic bacteriophages. In some embodiments, the bacteriophage or fungal virus formulation is part of a cosmetic formulation. The cosmetic formulation may also include one or more liposomes containing the first bacteriophage and the second bacteriophage. The cosmetic formulation may further include an anti-aging ingredient.

[0064] The phage isolates used in the formulation of phage functionally activated polymeric fabric (eg, nonwoven) surfaces are provided in Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0065] In some embodiments, the phage isolates used in the formulation of phage-activated polymer fabric (e.g., nonwoven) surfaces have a nucleic acid sequence that is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% similar to any one of the sequences listed in Table 2 or any one of SEQ ID NOs: 1-132. [Table 2-1] [Table 2-2] Table 2-3 Table 2-4 Table 2-5 Table 2-6 Table 2-7 Table 2-8 Table 2-9 Table 2-10 Table 2-11 Table 2-12 Table 2-13 Table 2-14 Table 2-15 Table 2-16 Table 2-17 Table 2-18 Table 2-19 [Table 2-20] [Table 2-21] [Table 2-22] [Table 2-23] [Table 2-24] [Table 2-25] [Table 2-26] [Table 2-27]

[0066] A polymeric nonwoven fabric for use in a personal care product is provided. The polymeric nonwoven fabric comprises a phage combination bound to the polymeric fabric, the phage combination comprising one or more sets of phages that infect one or more bacterial species. The one or more bacterial species are selected from a group of microorganisms including Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, Corynebacterium tuberculostearicum, or combinations thereof. The personal care product may be, for example, a face mask, a surgical face mask, a sanitary napkin, a tampon, a baby diaper, or an adult incontinence product.

[0067] In some embodiments, the personal care product is a surgical face mask with an inner sheet made of a nonwoven fabric, such as polypropylene, rayon / polyester blend, or polycaprolactone (PCL). Face masks wear on the face, increasing moisture and temperature on the skin. Bacteria, including Staphylococcus aureus, Staphylococcus argenteus, and Cutibacterium acnes, grow on the inner surface of the face mask as well as on the surface of the skin. The growing bacteria and irritated, moist skin promote the development of folliculitis, also known as maskne, a portmanteau of mask and acne. To reduce the growth of S. aureus / argenteus and C. acnes, phages infecting these species are attached to the nonwoven fabric by the following method: The surface of the nonwoven fabric is activated by imparting a positive charge via surface plasma treatment or corona discharge treatment. Phages are attached to the activated surface of nonwoven fabrics by spraying, misting, printing, or impregnating the fabric in a low-salt buffer (salt concentration less than 50 mM) containing at least 10 PFU / mL (PFU = plaque-forming units). The naturally negatively charged capsid of the phage binds to the charged, activated surface of the nonwoven fabric, making the positively charged phage tails prone to infecting and killing bacteria that come into contact with the surface of the nonwoven fabric incorporated into the face mask (see Figure 1). Alternatively, phage can be covalently attached to polypropylene or rayon / polyester nonwoven fabrics by surface activation chemistries such as EDC / NHS chemistry or by acidic surface activation, which covalently bonds amine groups on amino acid side chains on the phage capsid to activated carboxyl groups on the polymer surface.

[0068] In some embodiments, polymer nonwoven fabrics are used in adult incontinence diapers with a topsheet made from a nonwoven fabric, such as curved fibers (cellulose fibers, polyester fibers, polyethylene fibers, nylon fibers, polypropylene fibers, polylactic acid fibers, polyhydroxyalkanoate fibers, polyethylene terephthalate fibers, polypropylene carbonate fibers, or a mixture of these various fibers). When urine is absorbed by the diaper and retained against the skin, it increases moisture and pH, reducing the skin's natural moisture barrier and causing redness, skin bruising, and sometimes bleeding. Bacteria such as Enterococcus species and Escherichia coli proliferate, exacerbating incontinence-associated dermatitis (IAD). To reduce the growth of bacteria associated with IAD, phages targeting Enterococcus species and E. coli are attached to the polymer nonwoven topsheet of the diaper in a manner similar to that used for surgical face masks, and the surface is activated by providing a charge to bind to the negatively charged phage capsid or by creating active sites for covalent binding to the phage capsid.

[0069] Polymer nonwoven fabrics can be used in baby diapers with topsheets made from nonwoven fabrics such as polyester fibers, polyethylene fibers, polypropylene fibers, rayon / viscose fibers, polylactic acid fibers, acrylic, olefin, polyhydroxyalkanoate fibers, polyethylene terephthalate fibers, polypropylene carbonate fibers, or blends of these various fibers. Diaper dermatitis (DD) in infants is strongly associated with Candida albicans, Staphylococcus aureus / argenteus, and Enterococcus species. S. aureus / argenteus, in particular, produces exfoliative toxins that can exacerbate DD and show a strong statistical correlation with DD. Phages that infect S. aureus / argenteus and Enterococcus species can be attached to the polymer nonwoven topsheet of diapers in a manner similar to that used in surgical face masks.

[0070] Polymer nonwoven fabrics can be used in sanitary napkins with topsheets made from nonwoven fabrics such as polypropylene fibers, polyethylene fibers, polyethylene / polypropylene fibers, polylactic acid fibers, polyhydroxyalkanoate fibers, polyethylene terephthalate fibers, polypropylene carbonate fibers, or blends of these fibers. Although uncomfortable, napkin rash is not a life-threatening complication of wearing sanitary napkins. Sensitive skin around the vagina becomes irritated due to friction, increased moisture, elevated pH, and elevated temperature caused by the napkin. Skin occlusion also commonly promotes bacterial growth on the skin surface, which can contribute to napkin rash. Less frequently, but more critically, sanitary napkins can promote the growth of Staphylococcus aureus, Staphylococcus argenteus, or Streptococcus species, which can invade the vagina and reach the bloodstream. If they do reach the bloodstream, they can cause life-threatening device-associated menstrual toxic shock syndrome. Phages that infect S. aureus / argenteus and Streptococcus species can be attached to the polymeric nonwoven topsheet of a sanitary napkin in a manner similar to that of a surgical face mask. The phage-functionalized topsheet prevents the growth of S. aureus / argenteus and Streptococcus species in the napkin and on the skin surface, thus reducing the incidence of napkin rash and device-associated menstrual toxic shock syndrome.

[0071] In some embodiments, polymeric nonwoven fabrics are used in tampons with a topsheet made from a nonwoven fabric, such as polypropylene fibers, polyhydroxyalkanoate fibers, polyethylene fibers, polyethylene terephthalate fibers, polylactic acid fibers, polypropylene carbonate fibers, or a blend of these fibers. Tampons can promote the growth of Staphylococcus aureus, Staphylococcus argenteus, or Streptococcus species if not changed after 4 to 8 hours. The bacteria can reach the bloodstream, where they can cause life-threatening device-associated menstrual toxic shock syndrome. Phages that infect S. aureus, S. argenteus, and Streptococcus species can be attached to the polymeric nonwoven topsheet of sanitary napkins in a manner similar to that used in surgical face masks. The phage-functionalized topsheet prevents the growth of S. aureus / argenteus and Streptococcus species in the napkin and on the skin surface, thus reducing the incidence of device-associated menstrual toxic shock syndrome.

[0072] The phage isolates used in the formulation of phage functionally activated polymeric fabric (eg, nonwoven) surfaces are provided in Table 1.

[0073] 6.2.1. Formulation Bacteriophage preparations are described in WO2022198091A1, the entirety of which is incorporated herein by reference. The bacteriophage preparation or mixture may contain at least one bacteriophage capable of lysing a microorganism (e.g., bacteria) or inhibiting the growth of a microorganism (e.g., a microorganism found in a biological sample (e.g., skin, intestine, mouth) obtained from a subject), and in some cases may be useful in treating a health condition (e.g., a skin condition or disorder). The method may include determining or identifying the microorganism in the sample (e.g., identifying the host bacterium) and producing a bacteriophage mixture or bacteriophage preparation containing at least one bacteriophage capable of infecting the microorganism. The bacteriophage mixture or bacteriophage preparation can be administered to the subject from whom the sample was obtained. Thus, the methods provided herein may be useful in producing personalized preparations or mixtures for treating an individual's health condition. In some cases, a bacteriophage mixture or formulation includes at least two different bacteriophages, which may have the same or different host ranges. Combinations of such bacteriophages with designated host ranges can help prevent bacterial or microbial resistance.

[0074] The present disclosure also provides formulations for incorporating bacteriophages or bacteriophage mixtures into, for example, personal care products. The formulations may include cosmetic formulations of skin care formulations. The formulations may include any useful ingredients or compositions, such as excipients configured to stabilize one or more bacteriophages (e.g., maintain the viability of one or more bacteriophages over a period of time (e.g., at least one week)). The excipients may include substances for bulking solid, liquid, or gel formulations containing one or more bacteriophages. In some examples, the substances can provide therapeutic enhancements to one or more bacteriophages by, for example, enhancing solubility, decreasing or increasing dissolution, enhancing stability, increasing penetration into the skin or a portion thereof (e.g., the stratum corneum), increasing bacteriophage activity, etc. In some cases, the excipients may include one or more liposomes or lipophilic moieties (e.g., micelles, vesicles) capable of encapsulating one or more bacteriophages. Excipients or excipient substances can be used to alter the properties of a composition, such as viscosity. Substances can be used to alter the properties of a therapeutic agent, such as bioavailability, absorption, hydrophilicity, hydrophobicity, pharmacokinetics, etc. Excipients may include binders, anti-adhesives, coatings, disintegrants, glidants (e.g., silica gel, talc, magnesium carbonate), lubricants, preservatives, adsorbents, sweeteners, vehicles, or combinations thereof. For example, excipients may include powders, minerals, metals, sugars (e.g., monosaccharides or polysaccharides), sugar alcohols, naturally occurring polymers (e.g., cellulose, methylcellulose), synthetic polymers (e.g., polyethylene glycol or polyvinylpyrrolidone), alcohols, thickeners, starches, macromolecules (e.g., lipids, proteins, carbohydrates, nucleic acid molecules), etc.

[0075] The formulation may include additional ingredients for treating skin conditions (e.g., aging, acne). For example, the formulation may include one or more anti-aging ingredients, including, but not limited to, retinoids (vitamin A derivatives), niacinamide (vitamin B3), ascorbic acid (vitamin C), skin-active peptides, skin-active proteins or peptides (e.g., collagen, hyaluronic acid, or derivatives thereof), plant growth factors, such as kinetin, ubiquinone (coenzyme Q10), and the like. Alternatively, or in addition, the formulation may include one or more anti-inflammatory agents, such as antihistamines, salicylates, and the like. In some cases, the formulation may include one or more anti-acne agents, such as benzoyl peroxide, salicylic acid, probiotics, antibiotics, antifungals, and the like.

[0076] In some embodiments, the composition contains one or more bacteriophages formulated in a mixture. In some embodiments, the composition is formulated in a liquid, semi-liquid, solid, semi-solid, or powder form. In some embodiments, the composition is formulated to bind the phage to a surface-active nonwoven fabric and incorporated into a disposable personal care product. In some embodiments, the composition is formulated in a liquid for applying the composition to a surface-active nonwoven fabric.

[0077] In some embodiments, the composition contains one or more bacteriophages (eg, a cocktail of bacteriophages) formulated in culture medium.

[0078] Polymeric fabrics containing the phage combinations described herein, alone or incorporated into personal care products, can be maintained, stored, or shipped in cultures having a pH value between 1 and 14. Suitable pH values ​​are about pH 1, about pH 2, about pH 3, about pH 4, about pH 5, about pH 6, about pH 7, about pH 8, about pH 9, about pH 10, about pH 11, about pH 12, about pH 13, or about pH 14. By way of example, the pH value may be acidic or lower than pH 7 (e.g., pH 6.9, pH 6.5, pH 6, pH 5, pH 4, pH 3, pH 2, pH 1). By way of another example, the pH value may be alkaline or higher than pH 7 (e.g., pH 7.1, pH 7.5, pH 8, pH 9, pH 10, pH 11, pH 12, pH 13, or pH 14). By way of yet another example, the pH value may be closer to neutral (e.g., about pH 7, about pH 6 to about pH 8).

[0079] Polymeric fabrics bound with phage combinations or personal care products incorporating phage combinations or phage cocktails described herein can be packaged, stored, or shipped at about 4° C. to about 100° C., about 10° C. to about 80° C., about 15° C. to about 60° C., about 20° C. to about 40° C., or about 25° C. to about 35° C. In some embodiments, personal care products incorporating bacteriophages or bacteriophage cocktails are packaged, stored, or shipped at temperatures below 100° C., below 90° C., below 80° C., below 70° C., below 60° C., below 50° C., below 40° C., below 30° C., below 20° C., below 10° C., or below 4° C. In some embodiments, personal care products incorporating bacteriophages or bacteriophage cocktails are packaged, stored, or shipped at room temperature.

[0080] 6.2.2. Polymer Fabrics In some embodiments, the polymer fabric is a polymer matrix. As used herein, the term "polymer matrix" refers to a polymer that undergoes physicochemical changes in response to compounds produced or released by phage infection of microorganisms. The polymer may be degradable or non-degradable. Suitable polymers include, but are not limited to, polylactic acid, polycaprolactone, PHA, and PHB. The polymer matrix may comprise one type of polymer or a combination of polymers.

[0081] In some embodiments, the polymer fabric is a polymer matrix and comprises a composite of polymers. In some embodiments, the polymer matrix comprises two polymer layers. In some embodiments, the polymer matrix comprises a degradable polymer and / or a non-degradable polymer. The polymer matrix can be patterned at the nanometer scale, micrometer scale, or millimeter scale, or a combination thereof. Such patterning gives the polymer matrix a unique appearance, which may comprise a pure polymer or a polymer blended with a non-degradable polymer or non-polymer material. In some embodiments, the top layer comprises a degradable polymer and the bottom layer comprises a porous polymer.

[0082] In some embodiments, the polymer matrix is ​​a free-standing film of polymer, a blend of polymers, or a composite material. In some embodiments, the polymer matrix is ​​coated onto a carrier layer in the form of a film or coating.

[0083] In some embodiments, the polymer fabric is a nonwoven fabric. As used herein, the term "nonwoven fabric" refers to a fabric-like material made from short (short) and continuous (continuous) fibers bonded together by chemical, mechanical, thermal, or solvent treatment. Nonwoven fabrics are processed fabrics that can be single-use, limited-life, or highly durable. Nonwoven fabrics provide specific functions, such as absorbency, liquid repellency, elasticity, extensibility, softness, strength, flame retardancy, washability, cushioning, thermal insulation, sound insulation, filtration, bacterial barrier use, and sterility. Nonwoven fabrics can be used alone or as components of personal care products such as face masks, sanitary napkins, tampons, baby diapers, or adult incontinence products.

[0084] In some embodiments, the nonwoven fabric is treated by plasma surface treatment to produce a surface-active nonwoven fabric. For example, phage can be covalently or non-covalently bound to the surface-active nonwoven fabric using the methods described herein and those disclosed in U.S. Patent No. 9,277,751; Wang et al., Immobilization of active bacteriophages on polyhydroxyalkanoate surfaces. ACS Appl. Mater. Interfaces, 216, 8, 2, 1128-1138, each of which is incorporated herein by reference in its entirety.

[0085] In some embodiments, the phage is non-covalently bound to a surface-active nonwoven fabric incorporated into a disposable personal care product. The phage is bound such that the capsid is adsorbed to the surface of the nonwoven fabric by electrostatic charge, exposing the tail away from the surface of the fabric. In some embodiments, misting is used to bind the phage to the surface-active nonwoven fabric.

[0086] In some embodiments, phages are immobilized on a polymer fabric (e.g., a nonwoven fabric) by chemical bonding, physical adsorption, bulk mixing, or a combination thereof. One or more phages can be immobilized on a polymer fabric to enable the killing or suppression of the growth of multiple microbial strains. Methods for immobilizing phages on a polymer matrix are described in U.S. Patent No. 9,921,219, the entire contents of which are incorporated herein by reference. Suitable methods for immobilization include, but are not limited to, biotinylation of phage capsids for biotin-streptavidin affinity binding (Gervais et al., 2007; Edgar et al., 2006; Smelyanski et al., 2011); direct covalent attachment using n-hydroxysulfosuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide as linkers (Hermanson, 2008); and plasma-treated surface attachment (Pearson et al., 2013) (each incorporated herein by reference in its entirety).

[0087] 6.2.3. Personal care products In some embodiments, the phage combination is attached to a personal care product that comes into direct or chronic contact with the skin of a subject. Non-limiting examples of personal care products are face masks, sanitary napkins, tampons, baby diapers, adult incontinence products, cosmetics, cosmetic facial pads or wipes, fingernail wipes, or patient bath hygiene fabrics.

[0088] As described herein, the personal care product may be a face mask incorporating a phage combination. The face mask may be a surgical or non-surgical face mask. Such face masks typically have an inner sheet made of a nonwoven fabric, such as polypropylene, rayon / polyester blend, or polycaprolactone (PCL). Face masks cause wear on the face, increasing moisture and temperature on the skin. Bacteria, including Staphylococcus aureus, Staphylococcus argenteus, and Cutibacterium acnes, grow on the inner surface of the face mask as well as on the surface of the skin. The growing bacteria and irritated, moist skin promote the development of folliculitis, also known as maskne, a portmanteau of mask and acne. To reduce the growth of S. aureus, S. argenteus, and C. acnes, phages infecting these species are attached to the nonwoven fabric by the methods described herein (e.g., Example 4).

[0089] The personal care product may be an adult incontinence diaper incorporating the phage combination. The diaper may have a topsheet made of a nonwoven fabric, such as curved fibers, cellulose fibers, polyester fibers, polyethylene fibers, nylon fibers, polypropylene fibers, polylactic acid fibers, polyhydroxyalkanoate fibers, polyethylene terephthalate fibers, polypropylene carbonate fibers, or a mixture of these various fibers. When urine is absorbed by the diaper and retained against the skin, it increases moisture and pH, reducing the skin's natural moisture barrier and causing redness, skin bruising, and sometimes bleeding. Bacteria such as Enterococcus species and Escherichia coli grow, exacerbating incontinence-associated dermatitis (IAD). To reduce the growth of bacteria associated with IAD, phages targeting Enterococcus species and E. coli can be attached to the polymeric nonwoven topsheet of a diaper in a manner similar to that of a surgical face mask, activating the surface by providing a charge for binding to the negatively charged phage capsid or creating active sites for covalent binding to the phage capsid.

[0090] As another example, the personal care product may be a baby diaper incorporating a phage combination. The diaper may have a topsheet or outer surface made from a nonwoven fabric, such as polyester fibers, polyethylene fibers, polypropylene fibers, rayon / viscose fibers, polylactic acid fibers, acrylic, olefin, polyhydroxyalkanoate fibers, polyethylene terephthalate fibers, polypropylene carbonate fibers, or a blend of these various fibers. Diaper dermatitis (DD) in infants is strongly correlated with Candida albicans, Staphylococcus aureus, S. argenteus, and Enterococcus species. In particular, S. aureus and S. argenteus produce exfoliative toxins that can exacerbate DD and show a strong statistical correlation with DD. Phages that infect S. aureus, S. argenteus, and Enterococcus species can be attached to the polymeric nonwoven topsheet of the diaper in a manner similar to that of surgical face masks.

[0091] The personal care product may be a feminine care product, such as a sanitary napkin, incorporating the phage combination. The sanitary napkin may have a topsheet or outer surface made of a nonwoven fabric, such as polypropylene fibers, polyethylene fibers, polyethylene / polypropylene fibers, polylactic acid fibers, polyhydroxyalkanoate fibers, polyethylene terephthalate fibers, polypropylene carbonate fibers, or a mixture of these various fibers. Napkin rash, while unpleasant, is not a life-threatening complication of wearing sanitary napkins. Sensitive skin around the vagina becomes irritated due to friction, increased moisture, elevated pH, and elevated temperature caused by the napkin. Skin occlusion also commonly promotes bacterial growth on the skin surface, which can contribute to napkin rash. Less frequently, but more critically, sanitary napkins can promote the growth of Staphylococcus aureus, S. argenteus, or Streptococcus species, which can invade the vagina and reach the bloodstream. If they reach the bloodstream, they can cause life-threatening device-associated menstrual toxic shock syndrome. Phages that infect S. aureus, S. argenteus, and Streptococcus species can be attached to the polymeric nonwoven topsheet of a sanitary napkin in a manner similar to that of a surgical face mask. The phage-functionalized topsheet prevents the growth of S. aureus, S. argenteus, and Streptococcus species in the napkin and on the skin surface, thus reducing the incidence of napkin rash and device-associated menstrual toxic shock syndrome.

[0092] Another example of a personal care product is a tampon incorporating a phage combination. The tampon may have a topsheet or outer surface made from a nonwoven fabric, such as polypropylene fibers, polyhydroxyalkanoate fibers, polyethylene fibers, polyethylene terephthalate fibers, polylactic acid fibers, polypropylene carbonate fibers, or a mixture of these various fibers. If the tampon is not changed after a certain period of time (e.g., about 4 to about 8 hours), it can promote the growth of Staphylococcus aureus, S. argenteus, or Streptococcus species. The bacteria can reach the bloodstream. If they do, they can cause life-threatening device-associated menstrual toxic shock syndrome. Phages that infect S. aureus / argenteus and Streptococcus species can be attached to the polymeric nonwoven topsheet of a sanitary napkin in a manner similar to that of surgical face masks. The phage-functionalized topsheet prevents the growth of S. aureus, S. argenteus, and Streptococcus species in the napkin and on the skin surface, thus reducing the incidence of device-associated menstrual toxic shock syndrome.

[0093] 6.3. Method 6.3.1. Identification of Bacteriophage Combinations Bacteriophage selection: Bacteriophage can be selected for inclusion in mixture or formulation based on any useful characteristics.The characteristics that can be used for bacteriophage selection include, but are not limited to, non-lysogenic or non-integrated in the host genome, host range (e.g., specific to the host organism with minimal off-target binding), and stability in a given formulation.In some cases, bacteriophage combinations can be selected for a specific microbiota type or host range (e.g., to target a given bacterial strain, such as C. acnes), which can be combined to help prevent bacterial resistance.

[0094] Therefore, in a second aspect, the present disclosure provides a method for selecting phages to target the bacteria most commonly associated with contact dermatitis. In certain embodiments, the contact dermatitis is one or more of the following skin conditions: diaper rash, maskne, sanitary napkin rash, or incontinence-related rash.

[0095] In some embodiments, the method further comprises screening a skin sample from a subject or a population of subjects to identify bacteria that are commonly present and associated with contact dermatitis. Bacteriophage assays are described in WO2022198091A1, the entire contents of which are incorporated herein by reference.

[0096] Bacteriophage Assay: Screening of a sample can be performed to determine one or more agents (e.g., small molecules, bacteriophages, or biomolecules, such as proteins, peptides, lipids, carbohydrates, metabolites, or combinations thereof) that can inhibit the growth of or otherwise kill (e.g., by lysis) one or more microorganisms in the sample. In some examples, the one or more agents include one or more bacteriophages. For example, the one or more bacteriophages can be naturally occurring, recombinant, or synthetic bacteriophages that can lyse certain types of bacteria present on the skin sample. In such examples, screening of the sample can include obtaining one or more microorganisms from the sample, optionally culturing the one or more microorganisms, and exposing the one or more microorganisms to multiple bacteriophages to generate a microorganism-bacteriophage mixture. Growth, proliferation, or death of the microorganisms can be monitored over a period of time, and the microorganisms can be collected, stored, and / or sequenced. Sequencing (e.g., shotgun metagenomic sequencing) can provide information about bacteriophages present in or on microorganisms that may be capable of killing or inhibiting the growth of one or more microorganisms. Thus, such screening, for example, combining culturing microorganisms with bacteriophages and sequencing the mixture, can be useful in determining combinations of bacteriophages to eliminate specific microbial (e.g., bacterial) strains.

[0097] In some cases, screening may include sequencing one or more microorganisms isolated from the sample. For example, one or more microorganisms isolated from the sample can be (i) cultured to generate a microbial library and (ii) sequenced (e.g., by shotgun metagenomic sequencing) to generate a library of sequence reads containing sequences derived from the microorganisms present in the sample. The cultured microorganisms can then be further exposed to one or more bacteriophages, followed by isolation and sequencing, as described above. Sequences corresponding to one or more bacteriophages can be stored in a bacteriophage library (Table 1). Alternatively, or in addition, one or more bacteriophages can be stored and further screened, for example, using a bacteriophage host range assay, to determine bacterial strains that may be susceptible to one or more bacteriophages.

[0098] Bacteriophage Source: The one or more bacteriophages may be synthetic or naturally occurring. The one or more bacteriophages may be isolated or extracted from environmental sources, including, but not limited to, wastewater, soil suspensions, marine sediments, subsurface of the Earth, etc. In such cases, further processing of the environmental source may be carried out to purify or enrich the bacteriophage.

[0099] Bacteriophage Host Range Assay: Bacteriophage host range assays can be used to determine in vitro the host or set of host cells (e.g., bacterial cells) that are susceptible to a given bacteriophage. In some cases, bacteriophage host range assays are performed using one or more microorganisms isolated from a sample or multiple samples and one or more bacteriophages present in an environmental sample. For example, bacteria from one or more skin samples can be collected, and optionally, a portion of the collected bacteria can be sequenced to identify the bacterial type (e.g., bacterial species or variant). The collected bacteria can be cultured (e.g., on an agar plate), distributed onto a phage host range assay dish, and contacted with a viral fraction from an environmental source that may contain one or more bacteriophages (e.g., lytic bacteriophages). If desired, the collected bacteria can be pre-sorted or pre-treated to ensure that only one strain type or class of strain types is present per phage host range assay dish. If desired, the bacteria-bacteriophage mixture on the phage host range assay can be cultured. Bacteriophages that grow on or within the collected and cultured bacteria and / or lyse or prevent the growth of cultured bacteria on the assay can be isolated, optionally propagated, and stored as part of a bacteriophage library. Alternatively, or in addition, the bacteriophages can be sequenced. Such sequencing data can then be used to associate each isolated bacteriophage with a specific host organism. Thus, such assays can be useful in determining the identity and genome sequence of bacteriophages that can remove one or more microorganisms from a sample. If the identity of one or more microorganisms is also known (e.g., from sequencing), the host range of each bacteriophage can be determined. Thus, for a given target bacterial strain, appropriate bacteriophages can be selected for inclusion in a formulation designed to lyse the target bacterial strain.

[0100] 6.3.2. Binding of Bacteriophage Combinations to Polymer Fabrics Also disclosed herein are methods for attaching phages to polymeric fabrics that are incorporated into disposable personal care products. In some embodiments, the polymeric fabric is a nonwoven fabric.

[0101] The surface of a polymer fabric (e.g., a nonwoven fabric) is activated by providing a positive charge through surface plasma treatment or corona discharge treatment. Phages are attached to the activated surface of the polymer fabric (e.g., a nonwoven fabric) by spraying, misting, or printing. In certain embodiments, phage are attached to the activated surface of the polymer fabric by misting. The polymer fabric (e.g., a nonwoven fabric) is coated with at least 10 9 The phage can be impregnated in a low-salt buffer (salt concentration less than 50 mM) containing PFU / mL (PFU = plaque-forming units). Without being bound by any theory, the naturally negatively charged capsid of the phage binds to the charged and activated surface of the nonwoven fabric, making the positively charged phage tails more likely to infect and kill bacteria that come into contact with the surface of the polymer fabric (e.g., nonwoven fabric) incorporated into the face mask. Alternatively, phage can be covalently attached to polypropylene or rayon / polyester nonwoven fabrics by surface activation chemistries such as EDC / NHS chemistry, or by acidic surface activation, which covalently bonds amine groups on amino acid side chains on the phage capsid to activated carboxyl groups on the polymer surface.

[0102] An exemplary method for attaching bacteriophage combinations to polymer fabrics is shown in Figure 8, which describes a process that allows for the processing of nonwoven fabrics of various compositions impregnated with phages of various morphologies and host specificities after plasma treatment of the nonwoven fabric inline during the manufacturing process, as follows: (1) the nonwoven fabric is plasma treated inline in an automated manufacturing process (plasma treatment either in a case or in open air); (2) the surface-active nonwoven fabric can be impregnated on both surfaces using a dip tray (also called an impregnation tank or dip tank); and (3) the nonwoven fabric is gently dried using an air dryer at a temperature below 50°C, which does not denature the phage virions.

[0103] 6.3.3. Target As used herein, the term "subject" generally refers to an animal, such as a mammal (e.g., a human) or an avian (e.g., a bird), or other organism, such as a plant. For example, a subject may be a vertebrate, a mammal, a rodent (e.g., a mouse), a primate, a monkey, or a human.

[0104] Animals may include, but are not limited to, farm animals, sport animals, and pets. A subject may be a healthy or asymptomatic individual, an individual having or suspected of having a disease (e.g., a skin condition, cancer) or a predisposition to a disease, infection, health condition, and / or an individual in need of treatment or suspected of needing treatment. A subject may be a human. A subject may be a patient.

[0105] In some embodiments, the subject has acne. In some embodiments, the subject has one or more skin conditions, such as, but not limited to, acne, diaper rash, maskne, sanitary napkin rash, and incontinence-related rash, redness, eczema, rosacea, aging skin, or skin infection. The subject may have or be at risk of having one or more skin conditions caused by or associated with a particular microbiota subtype (e.g., may be susceptible to or prone to a condition). The subject may have one or more microbial variants detected in or living on the subject's skin.

[0106] According to the provided treatment methods, a bacteriophage mixture or bacteriophage preparation can be used to treat skin conditions that may be caused by or associated with a specific microbiota subtype (e.g., a specific microorganism, such as a bacterial strain, or a combination of microorganisms). For example, the preparations described herein can be used to treat acne, inflammation, redness, eczema, rosacea, enlarged hair follicle pore size, rough skin, increased transepidermal water loss, skin dehydration, skin discoloration (e.g., hyperpigmentation), and elasticity imbalances in the skin or a portion thereof (e.g., the stratum corneum, dermis, etc.). The preparation may include one or more bacteriophages that can lyse or inhibit the growth of one or more microorganisms associated with a skin condition (e.g., Cutibacterium acnes or Staphylococcus aureus, which may be associated with acne).

[0107] As described herein, the skin sample may contain one or more microorganisms associated with a skin condition (e.g., acne, eczema, redness, etc.). Thus, the bacteriophage mixture and bacteriophage formulation can be targeted to any species or variant of skin microorganisms that may be associated with a skin condition. For example, one or more bacteriophages may target bacteria, such as bacteria from the phyla Actinobacteria, Firmicutes, Proteobacteria, and Bacteroidetes. The one or more microorganisms may, in some examples, include Cutibacterium bacteria (e.g., C. acnes, C. namnatense, C. avidum), Staphylococcus bacteria (e.g., S. aureus, S. epidermidis, S. warneri, S. pyogenes, S. mitis), Corynebacterium bacteria, Acinelobacter bacteria (e.g., A. johnsonii), Pseudomonas bacteria (e.g., P. aeruginosa), other bacteria, or combinations thereof. In some cases, one or more bacteriophages can be targeted to fungal microorganisms, including, but not limited to, yeasts such as Candida albicans, Rhodotorula rubra, Torulopsis, and Trichosporon cutaneum, dermatophytes such as Microsporum gypseum and Trichophyton rubrum, and non-dermatophyte fungi such as Rhizopus stolonifer, Trichosporon cutaneum, Fusarium, Scopulariopsis brevicaulis, Curvularia, Alternaria alternata, Paecilomyces, Aspergillus flavus, and Penicillium. One or more bacteriophages can also target a combination of microorganisms; such combinations can be determined, for example, using a host range assay described elsewhere herein or as described in WO2022198091A, the entire contents of which are incorporated herein by reference.

[0108] Kit Also provided herein are kits for incorporating a phage combination bound to a synthetic polymer nonwoven fabric into a disposable personal care product used in close contact with the skin. Such kits may include a phage combination noncovalently bound to a polymer fabric (e.g., a synthetic polymer nonwoven fabric). The kit may include a phage combination in culture for binding the phage combination onto the polymer fabric (e.g., a synthetic polymer nonwoven fabric). Such kits may include instructions for binding the phage combination onto the polymer fabric. The kit may also include reagents such as, for example, stabilizers, buffers, preservatives, fixatives, or pH-balancing reagents. The kit may further include instructions for applying, storing, transporting, or treating the personal care product incorporating the polymer fabric.

[0109] 6.4 Treatment Regimen In the methods provided herein, the terms "treatment," "treating," and the like are used generally herein to refer to obtaining a desired pharmacological and / or physiological effect. In certain embodiments, the effect is therapeutic in terms of a partial or complete response to microbial growth, including inhibiting the growth of or preventing the regrowth of the microorganism, and / or reducing or eliminating one or more symptoms associated with contact dermatitis.

[0110] 6.4.1. Combination Administration In some embodiments of the provided method for treating one or more skin conditions, the phage combination is administered to a subject in combination with one or more skin care products or drugs.For example, the phage combination bound to a polymer fabric (e.g., nonwoven fabric) can be used in combination with skin lotion, skin cream, toner, skin cleanser, and / or skin wipe.In another example, the phage combination bound to a polymer fabric (e.g., nonwoven fabric) can be used in combination with medical procedures, such as surgery.In such an example, the polymer fabric (e.g., mesh wound dressing) containing the phage combination can be applied to a wound, wound, or surgical site to prevent bacterial growth or infection.

[0111] 6.4.1.1 Bacteriophage administration In various embodiments, the phage combination bound to a polymer fabric (e.g., a nonwoven fabric) is incorporated into a personal care product for administration to a subject. In some examples, the phage combination bound to a polymer fabric (e.g., a nonwoven fabric) is administered to a subject by applying the polymer fabric directly onto the subject's skin.

[0112] In some embodiments, the phage combination is administered to the subject by the subject, a physician, a medical professional, and / or a caregiver.

[0113] In certain embodiments, the phage combination is administered once, twice, three times, four times, or more daily. In certain embodiments, the phage combination is administered once daily. In certain embodiments, the personal care product incorporating the phage combination is administered once every two days. In certain embodiments, the personal care product incorporating the phage combination is administered once every three days. In certain embodiments, the personal care product incorporating the phage combination is administered once every four days. In certain embodiments, the personal care product incorporating the phage combination is administered once every five days. In certain embodiments, the personal care product incorporating the phage combination is administered once every six days. In certain embodiments, the personal care product incorporating the phage combination is administered once every week. In certain embodiments, the personal care product incorporating the phage combination is administered once every two weeks. In certain embodiments, the personal care product incorporating the phage combination is administered once every three weeks. In certain embodiments, the phage combination-incorporated personal care products are administered once per month. In certain embodiments, the phage combination-incorporated personal care products are administered as needed.

[0114] In some embodiments, the phage combination-incorporated personal care product is administered to the subject until the subject's skin condition (e.g., contact dermatitis) is partially or completely eliminated. In some embodiments, the phage combination-incorporated personal care product administered to the subject reduces one or more symptoms of contact dermatitis or skin infection and / or reduces body odor. [Example]

[0115] 7. Working Example 7.1. Example 1: Composition of Multiple Phages That Infect Bacteria on the Skin Surface Associated with Contact Dermatitis In this example, phages are identified that can lyse or kill one or more bacteria, including Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, and Corynebacterium tuberculostearicum. Combinations of phages that target or infect the bacteria most commonly associated with contact dermatitis are attached to plasma-surface-treated synthetic polymer nonwoven fabrics, including polylactic acid (PLA), polyester (PE), acrylic, olefin, polyhydroxyalkanoate (PHA), such as polyhydroxybutyrate (PHB), polyethylene terephthalate (PET), polypropylene (PP), and polypropylene carbonate (PPC). The surface-treated nonwoven fabric is incorporated into disposable personal care products that come in contact with the skin, such as face masks, sanitary napkins, tampons, baby diapers, adult incontinence products, or cosmetic facial pads or wipes, fingernail wipes, or patient bathing hygiene fabrics.

[0116] 7.2. Example 2: Selection of Phages to Target Bacteria Most Commonly Associated with Contact Dermatitis In this example, phages are selected that target or infect the bacteria described in Example 1.

[0117] These bacterial strains are cultured. Phages that infect these strains are isolated from environmental samples. The environmental sample is enriched for phages by adding enrichment broth and broth culture to the sample and incubating overnight. The enriched phage population is separated from the broth culture by sterile filtration. Phages are isolated from the enrichment broth filtrate by classical top agar plaque plating. The phage isolates are amplified on the target bacterial strain. DNA extracts from the isolates are sequenced to screen the phage genomes for genes associated with antibiotic resistance, virulence, lysogeny, or transduction. The screened phages are tested for host range and combined to maximize host range. Sets of phages that infect the same host species are tested in pairs to prevent bacterial regrowth in broth culture. A pair that prevents bacterial regrowth is selected. The selected pair may be a pair that does not occur together in nature. They can be selected to complement their effects on bacterial regrowth. Specific subunits of the genomes of the selected phage pairs are selected for compositions in which they are selected to prevent antibiotic resistance or to have various other beneficial properties, such as inhibiting bacterial growth, infecting and / or killing bacteria, and preventing bacterial regrowth.

[0118] 7.3. Example 3: Binding of Phages to Nonwoven Fabrics Incorporated into Disposable Personal Care Products In this example, phages targeting or infecting the bacteria described in Example 1 are attached to a polymer nonwoven fabric. The phages are concentrated from bacterial growth broth and resuspended in low-salt phage buffer by TFF diafiltration. A plasma surface treatment is applied to the nonwoven fabric, and the phage buffer is misted onto the surface-treated nonwoven fabric and allowed to dry at room temperature. The nonwoven fabric is incorporated into a disposable personal care product.

[0119] 7.4. Example 4: Preparation of activated polymer substrates for binding of phage combinations In this example, a nonwoven fabric is used. The surface of the nonwoven fabric is activated by providing a positive charge through surface plasma treatment or corona discharge treatment. 9 Phages are attached to the activated surface of the nonwoven fabric by spraying, misting, printing, or impregnating the fabric in a low-salt buffer (salt concentration less than 50 mM) containing PFU / mL (PFU = plaque-forming units). The naturally negatively charged capsid of the phage binds to the charged, activated surface of the nonwoven fabric, making the positively charged phage tail prone to infecting and killing bacteria that come into contact with the surface of the nonwoven fabric incorporated into the face mask.

[0120] Figure 1 illustrates the use of a composition comprising a phage described herein for use in a personal care product. A subject's skin harbors various bacterial species living commensally in or on the skin surface (1). Phages target specific types of bacteria (2) by recognizing phage receptors (3) displayed on the target host bacterial species. Note that non-targeted commensal skin microorganisms (4) remain undisrupted. Phage (5) displays a negatively charged capsid (6) and is capable of binding to a positively charged (7) activated polymer fabric surface (8). The polymer fabric is a nonwoven fabric. Phage (5) is a myovirus-tailed phage. Phage (9) is a podovirus-tailed phage, a different phage morphotype from myovirus-tailed phage. Upon phage binding, the polymer fabric (e.g., surface-activated nonwoven fabric) (8) is functionalized with immobilized phage (5) or (9).

[0121] Alternatively, phages can be covalently attached to polypropylene or rayon / polyester nonwoven fabrics by surface activation chemistries such as EDC / NHS chemistry or by acidic surface activation, which covalently binds amine groups on amino acid side chains on the phage capsid to activated carboxyl groups on the polymer surface.

[0122] In some cases, more than one type of phage (e.g., myovirus and podovirus, or phage that infects S. aureus and phage that infects E. coli) is attached to a fabric such as PET or PHB to target more than one type of bacteria that may be present in the area where the fabric is placed or comes into contact. Prior to this, it was not known whether it would be possible to attach phage such as myovirus or podovirus or both to nonwoven fabrics such as PET and PHB, and whether there would be challenges, for example, in preventing the attachment of both types of phage to the same piece of nonwoven fabric or the attachment of a mixture of phage to the nonwoven fabric. Furthermore, using phage treatment on personal care products is not simple, as such products may contain one or more chemicals or treatments, or various fabric properties that affect the attachment or use of phage.

[0123] 7.5. Example 5: Phage Binding to PET Nonwoven Fabric and Inhibition of Bacterial Growth In this example, nonwoven fabric was used. Phages were bound to the surface of the PET nonwoven fabric, and the inhibition of bacterial growth was measured.

[0124] Figures 2A-2C show that phage were able to bind to PET nonwoven fabric and inhibit E. coli growth. As shown in Figure 2A, a 1 cm2 plasma-treated PET nonwoven fabric square without phage binding was applied to an agar plate containing a bacterial lawn of Escherichia coli strain 11303. The clear border of the fabric square indicates that plasma-treated PET alone inhibited E. coli growth. However, no halo effect due to phage infection was observed.

[0125] Figure 2B shows that a 1 cm2 plasma-treated PET nonwoven square with E. coli myovirus T4 bound to it was applied to an agar plate containing a lawn of Escherichia coli strain 11303. There were clear halos extending from the PET square. These halos were plaques or lacunae formed by phage infection of E. coli. The data indicate that the phage successfully bound to the plasma-treated PET nonwoven square.

[0126] FIG. 2C is a repeat of FIG. 2B showing repeated phage binding to plasma-treated PET nonwoven squares.

[0127] Figures 4A-4C show that phage was able to bind to PET nonwoven fabric and inhibit the growth of Staphylococcus aureus. Figure 4A shows that a 1 cm2 plasma-treated PET nonwoven fabric square without phage binding was applied to an agar plate containing a bacterial lawn of Staphylococcus aureus strain 100311. The clear border of the fabric square indicates that plasma-treated PET alone inhibited the growth of S. aureus. However, no halo effect due to phage infection was observed.

[0128] Figure 4B shows that a 1 cm2 plasma-treated PET nonwoven square bound with the S. aureus podovirus Rosenblumvirus P00474 was applied to an agar plate containing a lawn of Staphylococcus aureus strain 100311. There were clear halos extending from the PET square. These halos were plaques or lacunae formed by phage infection of S. aureus. In support of Figure 1, the E. coli myovirus T4 successfully bound to the plasma-treated PET nonwoven square and inhibited E. coli growth on a parallel solid surface such as an agar plate.

[0129] FIG. 4C is a repeat of FIG. 4B showing repeated phage binding to plasma-treated PET nonwoven squares.

[0130] Taken together, the data show for the first time that phages can bind to and persist on PET polymer nonwovens and successfully inhibit the growth of multiple different bacteria.

[0131] 7.6. Example 6: Phage Binding to PHB Nonwoven Fabric and Inhibition of Bacterial Growth In this example, a nonwoven fabric was used. Phages were bound to the surface of the PHB nonwoven fabric, inhibiting bacterial growth.

[0132] Figures 3A-3C show that phages were able to bind to PHB nonwoven fabric and inhibit the growth of E. coli. As shown in Figure 3A, a 1 cm square of plasma-treated PHB nonwoven fabric without phage binding was applied to an agar plate containing a lawn of Escherichia coli strain 11303. No growth inhibition of E. coli due to phage infection was measured.

[0133] Figure 3B shows that 1 cm2 plasma-treated PHB nonwoven squares with E. coli myovirus T4 bound were applied to an agar plate containing a lawn of Escherichia coli strain 11303. There were clear halos extending from the PHB squares. These halos were plaques or lacunae formed by phage infection of E. coli. The data indicate that the phage successfully bound to the plasma-treated PHB nonwoven squares.

[0134] FIG. 3C is a repeat of FIG. 3B showing repeated phage binding to plasma-treated PHB nonwoven squares.

[0135] Figures 5A-5C show that phage was able to bind to PHB nonwoven fabric and inhibit the growth of Staphylococcus aureus. As shown in Figure 5A, a 1 cm square of plasma-treated PHB nonwoven fabric without phage binding was applied to an agar plate containing a lawn of Staphylococcus aureus strain 100311. No growth inhibition of S. aureus due to phage infection was measured.

[0136] Figure 5B shows the application of 1 cm2 plasma-treated PHB nonwoven squares with bound S. aureus podovirus Rosenblumvirus P00474 to an agar plate containing a lawn of Staphylococcus aureus strain 100311. There were clear halos extending from the PHB squares. These halos were plaques or lacunae formed by phage infection of S. aureus. Supporting Figure 1, the S. aureus podovirus successfully bound to the plasma-treated PHB nonwoven squares and inhibited the growth of S. aureus on parallel solid surfaces such as agar plates.

[0137] FIG. 5C is a repeat of B showing repeated phage binding to plasma-treated PHB nonwoven squares.

[0138] Taken together, the data show that phages were able to bind and persist on PHB polymer nonwovens and successfully inhibited the growth of several different bacteria.

[0139] 7.7. Example 7: Effect of T4 Myovirus Phage Bound to Nonwoven Fabric on Bacterial Growth In this example, nonwoven fabrics were used. T4 myovirus phage was bound to the surface of PET or PHB nonwoven fabrics, and bacterial growth was monitored.

[0140] Figure 6 shows kinetic growth curves for Escherichia coli strain 11303 demonstrating the effect of T4 myovirus phage binding to plasma-treated PET and PHB nonwovens. The dashed lines represent E. coli growth curves for 1) no phage infection, 2) phage infection at a multiplicity of infection (moi) of 0.001, 3) phage infection at a moi of 0.01, and 4) phage infection at a moi of 0.1. These were compared to the E. coli growth curves plotted as solid lines representing 1 cm2 plasma-treated nonwoven squares of either PET or PHB to which phage was bound. Phage bound to surfaces such as PET or PHB were not as readily available for bacterial infection as phage added in suspension to bacterial cultures. Nevertheless, phages bound to PET or PHB nonwoven squares inhibited E. coli growth better than a phage suspension at an moi of 0.001 and equally, if not more, efficient than a phage suspension at an moi of 0.1. E. coli myovirus phage T4 successfully bound to various types of plasma-treated nonwoven fabrics and inhibited E. coli growth.

[0141] 7.8. Example 8: Effect of podovirus phages bound to nonwoven fabric on bacterial growth In this example, nonwoven fabrics were used. S. aureus podovirus Rosenblumvirus P00474 phage was bound to the surface of PET or PHB nonwoven fabrics, and bacterial growth was monitored.

[0142] Figure 7 shows the kinetic growth curves of Staphylococcus aureus strain 100311, demonstrating the effect of S. aureus podovirus Rosenblumvirus PHP00474 bound to plasma-treated PET and PHB nonwovens. The dashed lines indicate the S. aureus growth curves for 1) no phage infection, 2) phage infection at an moi of 0.01, and 4) phage infection at an moi of 0.1. These were measured by applying 1 cm of either phage-bound PET or PHB to the nonwoven fabric. 2The S. aureus growth curve plotted with a solid line representing the plasma-treated nonwoven squares was compared to that of the S. aureus growth curve plotted with a solid line representing the plasma-treated nonwoven squares. Phages bound to surfaces such as PET or PHB were not as readily available for bacterial infection as phage added in suspension to bacterial cultures. Nevertheless, phage bound to PET nonwoven squares inhibited S. aureus growth as well as a phage suspension at an moi of 0.01. Phage bound to PHB nonwoven squares inhibited S. aureus growth as well as a phage suspension at an moi of 0.1. S. aureus podovirus successfully bound to various types of plasma-treated nonwovens. S. aureus podovirus can inhibit S. aureus growth.

[0143] 7.9. Example 9: Effect of Siphovirus Phage Bound to Nonwoven Fabric on Bacterial Growth In this example, nonwoven fabrics are used. Siphovirus phages are bound to the surface of PET or PHB nonwoven fabrics, and bacterial growth is monitored.

[0144] The growth of Siphovirus-infected bacteria was tested under four conditions: 1) no phage infection, 2) phage infection at an moi of 0.01, and 4) phage infection at an moi of 0.1. Phages bound to surfaces such as PET or PHB nonwoven squares inhibited the growth of Siphovirus-infected bacteria. Siphovirus was successfully bound to various types of plasma-treated nonwoven fabrics, and Siphovirus was able to inhibit the growth of target bacteria.

[0145] 7.10. Example 10: Surface modification of nonwoven fabric In this example, nonwoven fabric was used. 2 cm x 2 cm squares of PET or PHB fabric were treated with oxygen plasma. Plasma treatment was performed by reactive ion etching (μEtch) in a cylindrical chamber with a diameter of 30 cm. A uniform glow discharge was created in a copper wire coil wrapped around the chamber and connected to a radio frequency (RF) generator. Commercially available oxygen gas (99.993%, Praxair, Edmonton, Canada) was introduced into the chamber. Samples of PET or PHB polymer fabric were treated in the plasma at a floating potential for 1 minute at 100 W RF power, 25% oxygen concentration, and a vacuum of 100 mTorr.

[0146] 7.11. Example 11: Phage immobilization on nonwoven fabric In this example, nonwoven fabrics were used. Adsorption to untreated and plasma-treated (PT) fabrics. The binding method was carried out in PBS buffer at pH 7.5. Oxygen plasma-treated PET or PHB fabrics and untreated PET or PHB fabrics measuring 2 cm x 2 cm were immersed in a 125 mL shake flask containing 20 mL of myovirus or podovirus suspension (2 x 10 pfu / mL) and mixed at 4 °C and 60 rpm for 2 hours. After exposure to phage, the films were washed seven times in PBS buffer to remove any unbound or loosely bound phage. All washing steps were carried out at room temperature.

[0147] 7.12. Example 12: Preparation of Surface-Activated Nonwoven Fabric In this example, a nonwoven fabric was used.

[0148] This example illustrates the feasibility of in-line phage treatment of nonwoven fabrics during manufacturing. As shown in Figure 8, the described process allows for the treatment of nonwoven fabrics of various compositions impregnated with phage of various morphologies and host specificities after plasma treatment of the nonwoven fabric in-line during the manufacturing process, as follows: (1) the nonwoven fabric is plasma treated in-line in an automated manufacturing process (plasma treatment either in a case or in open air); (2) the surface-active nonwoven fabric can be impregnated on both surfaces using a dip tray (also called an impregnation tank or dip tank); and (3) the nonwoven fabric is gently dried using an air dryer at a temperature below 50 °C, which does not denature the phage virions.

[0149] Alternatively, the process is as follows: (1) the nonwoven fabric is plasma treated in-line in an automated manufacturing process (plasma treatment either in a case or in open air); (2) alternatively, the nonwoven fabric can be impregnated on only one surface using a spray-on or misting device; and (3) the nonwoven fabric is gently dried using an air dryer at a temperature below 50°C, which does not denature the phage virions.

[0150] 8. Equivalents and Incorporation by Reference While the present invention has been particularly shown and described with reference to preferred and various alternative embodiments, it will be understood by those skilled in the relevant art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.

[0151] All references, issued patents and patent applications cited in the body of this specification are hereby incorporated by reference in their entirety for all purposes.

Claims

1. A composition comprising a phage combination bound to a polymer fabric, the phage combination comprises one or more sets of phages that infect one or more bacterial species; the phage is selected from the family Myoviridae, Podoviridae, and Siphoviridae; The composition, wherein the polymeric fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

2. 2. The composition of claim 1, wherein the phage is of the genus Pahexavirus or Rosenblumvirus.

3. 3. The composition of claim 1 or claim 2, wherein the phage is a tailed virus.

4. The composition of any one of claims 1 to 3, wherein the phage is at least one of Myoviridae or Podoviridae.

5. The composition of claim 4 , wherein the phage is a T4 myovirus.

6. The composition of claim 5 , wherein the phage is a podovirus.

7. 7. The composition of claim 1, wherein the one or more sets of phages infect at least one common target bacterial species.

8. each set of phages comprises at least 2, 3, 4, 5 or more phages selected from one or more phage libraries; The composition of any one of claims 1 to 7, wherein the one or more phage libraries comprise phages from one or more genera.

9. The composition of claim 1 , wherein the phage combination is non-covalently or covalently bound to the polymer fabric.

10. The composition of claim 9 , wherein the polymeric fabric is a synthetic nonwoven fabric.

11. 11. The composition of any one of claims 1 to 10, wherein the phage combination is deposited on at least one material that forms at least part of a personal care product.

12. 12. The composition of claim 11, wherein the personal care product is selected from the group consisting of a face mask, a sanitary napkin, a tampon, a baby diaper, an adult incontinence product, or a cosmetic facial pad or wipe, a fingernail wipe, or a patient bathing hygiene fabric.

13. 13. The composition of any one of claims 1 to 12, wherein the bacterial species is commonly associated with contact dermatitis.

14. The composition of claim 13, wherein the bacterial species is selected from a skin microbiome biobank.

15. 15. The composition of claim 14, wherein the bacterial species is derived from skin microbiota isolated from a biological sample of a subject.

16. 16. The composition of any one of claims 1 to 15, wherein the one or more bacterial species are selected from a group of microorganisms comprising Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, Corynebacterium tuberculostearicum, or a combination thereof.

17. 17. The composition of any one of claims 1 to 16, wherein the phage is a phage that infects Escherichia coli.

18. 17. The composition of claims 1 to 16, wherein the phage is a phage that infects Staphylococcus aureus.

19. 19. The composition of any one of claims 1 to 18, wherein the polymer fabric further comprises a fabric selected from the group consisting of polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, polycaprolactone (PCL) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

20. A composition comprising a phage combination bound to a polymer fabric, the phage combination comprises one or more sets of Caudaviridae phages; The composition, wherein the polymer fabric is a polyethylene terephthalate (PET) nonwoven fabric.

21. A composition comprising a phage combination bound to a polymer fabric, the phage combination comprises at least a phage that infects Staphylococcus aureus; The composition, wherein the polymeric fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

22. A composition comprising a phage combination bound to a polymer fabric, the phage combination comprises at least a phage that infects Escherichia coli; The composition, wherein the polymeric fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

23. A composition comprising a phage combination bound to a polymer fabric, the phage combination comprises one or more sets of Caudaviridae phages; The composition, wherein the polymer fabric is selected from the group consisting of polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, polycaprolactone (PCL) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

24. A composition comprising a phage combination bound to a polymeric fabric, said phage combination comprising one or more sets of phages that infect one or more bacterial species.

25. 25. The composition of any one of claims 20 to 24, wherein the phage is of the genus Pahexavirus or Rosenblumvirus.

26. 26. The composition of any one of claims 20 to 25, wherein the phage is selected from Myoviridae, Podoviridae, and Siphoviridae.

27. 27. The composition of any one of claims 20 to 26, wherein the phage is a tailed virus.

28. 28. The composition of any one of claims 20 to 27, wherein the phage is at least one of Myoviridae or Podoviridae.

29. 29. The composition of claim 28, wherein the phage is a T4 myovirus.

30. 29. The composition of claim 28, wherein the phage is a podovirus.

31. 31. The composition of any one of claims 20 to 30, wherein the one or more sets of phages infect at least one common target bacterial species.

32. each set of phages comprises at least 2, 3, 4, 5 or more phages selected from one or more phage libraries; 32. The composition of any one of claims 20 to 31, wherein the one or more phage libraries comprise phages from one or more genera.

33. 33. The composition of any one of claims 20 to 32, wherein the phage combination is non-covalently or covalently bound to the polymer fabric.

34. 25. The composition of claim 24, wherein the polymeric fabric is a synthetic nonwoven fabric.

35. 35. The composition of any one of claims 20 to 34, wherein the phage combination is deposited on at least one material that forms at least a part of a personal care product that comes into contact with human skin.

36. 36. The composition of claim 35, wherein the personal care product is selected from the group consisting of a face mask, a sanitary napkin, a tampon, a baby diaper, an adult incontinence product, or a cosmetic facial pad or wipe, a fingernail wipe, or a patient bathing sanitary fabric.

37. 37. The composition of any one of claims 20 to 36, wherein the bacterial species is commonly associated with contact dermatitis.

38. 38. The composition of claim 37, wherein the bacterial species is selected from a skin microbiome biobank.

39. 38. The composition of claim 37, wherein the bacterial species is derived from skin microbiota isolated from a biological sample of a subject.

40. 40. The composition of any one of claims 20 and 23-39, wherein the one or more bacterial species are selected from a group of microorganisms comprising Staphylococcus aureus, Staphylococcus argenteus, Streptococcus pyogenes, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecalis, Cutibacterium acnes, Corynebacterium tuberculostearicum, or a combination thereof.

41. 41. The composition of any one of claims 20 and 21 to 40, wherein the phage combination further comprises a phage that infects at least Escherichia coli.

42. 41. The composition of claims 20 and 22-40, wherein the phage combination further comprises a phage that infects at least Staphylococcus aureus.

43. 43. The composition of any one of claims 20 to 42, wherein the polymer fabric is selected from the group consisting of polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, polycaprolactone (PCL) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

44. 44. A method for selecting a phage combination according to any one of claims 1 to 43.

45. 44. A method of binding a phage combination according to any one of claims 1 to 43 to a polymer nonwoven fabric.

46. 46. ​​The method of claim 45, wherein the binding of the phage combination is achieved by impregnating phage capsids onto the surface of the polymer nonwoven fabric.

47. 47. The method of claim 46, wherein the phage capsid is absorbed onto the surface of the polymeric fabric by electrostatic charge.

48. 48. The method of claim 47, wherein the tail is exposed from the fabric.

49. 49. The method of any one of claims 44 to 48, wherein the binding of the phage combination is achieved by mist spraying, chemical binding, physical adsorption, bulk mixing, plasma-treated surface binding, or a combination thereof.

50. 50. The method of any one of claims 44 to 49, wherein binding of the phage combination is achieved by plasma-treated surface binding.

51. 1. A method for preparing a personal care product, comprising: (a) contacting a polymeric nonwoven fabric with a composition comprising a phage combination; Including, the phage combination comprises one or more sets of phages of Caudavicetes phages that infect one or more bacterial species; The method, wherein the polymer nonwoven fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric.

52. 1. A method for preparing a personal care product, comprising: (a) contacting a polymeric nonwoven fabric with a plasma composition to produce a plasma-treated nonwoven fabric, wherein the polymeric nonwoven fabric is selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric; (b) immobilizing a phage combination comprising at least one or more sets of Caudavicetes phages on the polymer nonwoven fabric, wherein the phage combination comprises one or more sets of phages that infect one or more bacterial species; and (c) incorporating the polymeric nonwoven fabric into the personal care product. A method comprising:

53. A polymeric nonwoven fabric comprising a phage combination bound to a polymeric fabric, the phage combination comprises one or more sets of Caudaviridae phages that infect one or more bacterial species; The polymer fabric is selected from polylactic acid (PLA) nonwoven fabric, polyester (PE) nonwoven fabric, acrylic nonwoven fabric, olefin nonwoven fabric, polyhydroxyalkanoate (PHA) nonwoven fabric, polyhydroxybutyrate (PHB) nonwoven fabric, polyethylene terephthalate (PET) nonwoven fabric, polypropylene (PP) nonwoven fabric, and polypropylene carbonate (PPC) nonwoven fabric.

54. Manufacture of a personal care product for reducing contact dermatitis, reducing skin infections, and / or reducing body odor, the personal care product comprising a polymeric nonwoven fabric; the polymer nonwoven fabric comprises a phage combination bound to a nonwoven fabric selected from at least one of a polyhydroxybutyrate (PHB) nonwoven fabric or a polyethylene terephthalate (PET) nonwoven fabric; The phage combination comprises one or more sets of Caudoviricetes phages that infect one or more bacterial species.

55. 55. The personal care product of claim 54, selected from the group consisting of a face mask, a sanitary napkin, a tampon, a baby diaper, an adult incontinence product, or a cosmetic facial pad or wipe, a fingernail wipe, or a patient bathing sanitary fabric.

56. 55. A phage of the composition of any one of claims 1 to 43, the method of any one of claims 44 to 52, or the polymeric nonwoven fabric of claim 52, or the personal care product of claims 53 and 54, wherein the phage comprises a nucleic acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any one of SEQ ID NOs: 1 to 132.

57. 55. A phage of the composition of any one of claims 1 to 43, the method of any one of claims 44 to 52, or the polymeric nonwoven fabric of claim 52, or the personal care product of claims 53 and 54, wherein the phage comprises a nucleic acid sequence having at least 95% identity to any one of SEQ ID NOs: 1 to 132.

58. 55. A phage of a composition of any one of claims 1 to 43, a phage of a method of any one of claims 44 to 52, or a phage of a polymeric nonwoven fabric of claim 52, or a phage of a personal care product of claims 53 and 54, wherein the phage comprises a nucleic acid sequence having the sequence of any one of SEQ ID NOs: 1 to 132.

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