Strain and composition

Abstract

The present invention relates to strains and compositions. The present invention provides a bacterial strain having at least 95% genetic homology with one or more bacterial strains selected from the group consisting of: Lactobacillus plantarum LB244R, preserved at DSM 32996; the invention relates to lactobacillus plantarum LB356R, which is preserved in DSM 33094. The invention discloses lactobacillus plantarum LB316R, which is preserved in DSM 33091. The lactobacillus plantarum LB312R is preserved in the form of DSM 33098; the lactobacillus casei LB113R is preserved in the form of DSM 32907; the lactobacillus casei LB116R is preserved in the form of DSM 32908, and the lactobacillus casei LB116R is preserved in the The enterococcus faecium LB276R is preserved in the form of DSM 32997; the leuconostoc mesenteroides LB349R is preserved in the form of DSM 33093; the invention relates to Weissella virida LB10G, which is preserved in the form of DSM 32906. The invention also provides compositions comprising the one or more bacterial strains.

Description

[0001] This application is a divisional application of the invention patent application with the application date of August 8, 2019, the invention name of which is “Strains, Compositions and Methods of Use” and the application number is 2019800878642 (international application number is PCT / EP2019 / 071348). Technical Field

[0002] The present invention relates to novel bacterial strains. In particular, the present invention relates to novel bacterial strains for treating; alleviating; inhibiting; preventing and / or preventing the growth of pathogenic microorganisms. Background Art

[0003] The Gram-positive bacterium Staphylococcus aureus is one of the most common human pathogens. It is particularly common in the human nasal cavity, where it is found intermittently or permanently in approximately 50% of the population. Although S. aureus colonization is asymptomatic in most cases, the pathogen can cause infection when the skin's protective barrier function is disrupted. Consequently, staphylococcal infections are often associated with skin conditions such as dermatitis, eczema, carbuncles, cellulitis, rosacea, psoriasis, diaper rash, impetigo, and wounds.

[0004] An example of a skin infection is atopic dermatitis, a chronic or chronically relapsing inflammatory skin disease caused by a complex interplay of environmental, immune, genetic, and pharmacological factors.

[0005] All of these genetic and environmental factors contribute to the development of the following traits.

[0006] 1: Abnormal colonization with pathogenic microorganisms (e.g., Staphylococcus aureus) (compared with Staphylococcus epidermidis in normal individuals), which subsequently increases the patient's susceptibility to skin infections; in addition, Staphylococcus aureus produces enterotoxins that induce the production of enterotoxin-specific IgE, leading to greater T cell recruitment;

[0007] 2: Participate in the initial stage of the disease, thereby increasing the production of immunoglobulin E (IgE);

[0008] 3: Skin barrier dysfunction or dry skin due to abnormal lipid metabolism and / or epidermal structural protein formation; and

[0009] 4: The imbalance of the autonomic nervous system affects the patient's physical and mental health, followed by an increase in the production of mediators by various inflammatory cells.

[0010] Treatment of atopic dermatitis is typically with topical moisturizers / emollients or corticosteroids as first-line therapy, followed by topical calcineurin inhibitors as second-line therapy for patients not controlled with first-line therapy.

[0011] Currently, there are no topical treatments for patients with mild to moderate atopic dermatitis that are free of significant side effects such as localized thinning of the skin at the application site, worsening or risk of acne in patients, and burning sensations.

[0012] Unmet needs remain for patients with atopic dermatitis. Topical treatments still cause patients to suffer from well-known side effects such as itching. Scratching only worsens symptoms, leading to lichenification, excoriation, and skin barrier disruption, creating a vicious cycle for patients and increasing the risk of skin infections.

[0013] The most difficult unmet need yet to be addressed is the prevention and treatment of skin infections caused by Staphylococcus aureus, particularly those that are resistant to antibiotics, to avoid additional antibiotic resistance.

[0014] Even many different pathogenic microorganisms have been discovered, which may include bacterial microorganisms, viral microorganisms, fungal microorganisms, parasitic microorganisms and algal microorganisms, and the Gram-positive bacterium Staphylococcus aureus is one of the most common human pathogens.

[0015] With the discovery of penicillin in 1928 and its large-scale production in the early 1940s, most S. aureus infections were treatable without any serious complications. However, clinicians soon observed the emergence of penicillin-resistant S. aureus strains, primarily due to the expression of β-lactamases, enzymes that destroy the β-lactam ring structure in penicillins and cephalosporins (β-lactam antibiotics), thereby destroying their antibacterial activity. Methicillin, a new penicillin analog resistant to β-lactamases, was introduced in 1959 and initially showed efficacy against penicillin-resistant S. aureus strains. However, this success was short-lived, as the first methicillin-resistant S. aureus (MRSA) strain was identified in the laboratory in 1961, and the first clinical case of MRSA was observed in 1968.

[0016] MRSA infections primarily occur in hospital settings, and MRSA is currently one of the most common nosocomial pathogens and, therefore, a major cause of various hospital-acquired infections (HAIs). Since the 1990s, a new type of MRSA, termed community-associated MRSA (CA-MRSA), has emerged. CA-MRSA is not only genetically distinguishable from healthcare-associated MRSA (HA-MRSA) strains, but also exhibits different virulence and antibiotic resistance patterns. Recently, crossover between different CA-MRSA and HA-MRSA strains has been observed, which often makes it difficult to determine the source of the infecting MRSA strain.

[0017] HA-MRSA infection is associated with major clinical complications and frequently occurs in patients on ventilators, as well as those with surgical site wounds or surgical implants or requiring surgical wound drainage. Furthermore, HA-MRSA infection frequently leads to nosocomial pneumonia or bacteremia, which are associated with significant morbidity and mortality.

[0018] Because S. aureus infects not only humans but also other mammals, infected livestock and pets have become another source of transmission. Livestock-associated MRSA (LA-MRSA) has been identified, primarily in pigs, with colonization rates varying widely, from 10% to 80%, but LA-MRSA has also been found in ruminants and poultry.

[0019] LA-MRSA is a growing problem in the livestock industry. Objects such as stables, corrals, animals, farmers, their family members, stable owners and visitors, as well as slaughterhouses and animal transport vehicles can be contaminated with LA-MRSA. These objects can harbor LA-MRSA and be a source of transfer of LA-MRSA between objects, leading to the spread of LA-MRSA between healthy carriers. There is also the risk of LA-MRSA being transmitted by carriers to hospitals or nurseries, to patients with inflammatory skin diseases such as eczema and atopic dermatitis, carbuncles, cellulitis, rosacea, psoriasis, wounds, and burns, or to humans, including children and infants, causing diaper rash and impetigo caused by MRSA.

[0020] Therefore, it would be advantageous to improve the treatment of pathogenic microorganisms such as MRSA, in particular more effective and / or reliable treatments of pathogenic microorganisms such as MRSA, which do not have the disadvantages of currently available treatments, such as inflammatory skin diseases, such as eczema and atopic dermatitis, carbuncles, cellulitis, rosacea, psoriasis, wounds and burns, or diaper rash and impetigo in humans, including children and infants; and which do not induce the development of further resistance (e.g., antibiotic resistance) in the pathogenic microorganisms. Summary of the Invention

[0021] Thus, one object of the present invention relates to microorganisms, compositions and methods for treating pathogenic bacterial infections in subjects, such as mammals.

[0022] In particular, the present invention aims to provide a microorganism, composition, and method for treating pathogenic bacterial infections in subjects, such as mammals, that addresses the aforementioned prior art issues of skin infections, skin diseases, morbidity, mortality from infection, and antibiotic resistance. Thus, one aspect of the present invention relates to bacterial strains having at least 95% genetic homology to one or more bacterial strains selected from the group consisting of:

[0023] Weissella viridescens LB10G, deposited as DSM 32906;

[0024] Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0025] Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0026] Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0027] Enterococcus faecium LB276R, deposited as DSM 32997;

[0028] Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0029] -Leuconostoc mesenteriodes LB341R;

[0030] Leuconostoc mesenteriodes LB349R, deposited as DSM 33093;

[0031] Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0032] Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0033] Therefore, another aspect of the present invention relates to bacterial strains having at least 95% genetic homology to one or more bacterial strains selected from the group consisting of:

[0034] Weissella viridis LB10G, deposited as DSM 32906;

[0035] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0036] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0037] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0038] Enterococcus faecium LB276R, deposited as DSM 32997;

[0039] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0040] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0041] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0042] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0043] Another aspect of the present invention relates to a composition comprising one or more bacterial strains according to the invention.

[0044] Yet another aspect of the present invention relates to a composition of the present invention for use in treating, alleviating, inhibiting, preventing and / or preventing the growth of pathogenic microorganisms. DETAILED DESCRIPTION

[0045] The present invention relates to probiotics and compositions for preventing or treating infections caused by pathogenic microorganisms, such as staphylococcal infections. The present invention also relates to compositions and novel microbial strains capable of inhibiting the growth of pathogenic microorganisms, such as MRSA.

[0046] MRSA may be spread from one subject to another through direct contact with a carrier, such as an infected surface or another infected person, such as shared personal items that have come into contact with infected skin, or contact with contaminated objects, including the skin, nasal cavity, animals, surfaces, or objects.

[0047] Despite improvements in treatment, invasive MRSA infections, especially those that begin in community or livestock settings, remain problematic and impose a significant economic burden on healthcare systems, in addition to significant discomfort for patients. Similarly, in the European Union, MRSA accounts for 44% of all healthcare-associated infections (HAIs), 22% of attributable excess deaths, and 41% of excess HAI-related hospital stays. MRSA is a serious public health problem in Japan, which has one of the highest crude MRSA prevalence rates among all strains of Staphylococcus aureus in the world.

[0048] The present invention provides novel microbial strains and novel compositions that can inhibit the growth of Staphylococci without promoting the further development of antibiotic resistance.

[0049] Therefore, a preferred embodiment of the present invention relates to bacterial strains having at least 95% genetic homology to one or more bacterial strains selected from the group consisting of:

[0050] Weissella viridis LB10G, deposited as DSM 32906;

[0051] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0052] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0053] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0054] Enterococcus faecium LB276R, deposited as DSM 32997;

[0055] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0056] - Leuconostoc mesenteroides LB341R;

[0057] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0058] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0059] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0060] Therefore, another preferred embodiment of the present invention relates to bacterial strains having at least 95% genetic homology to one or more bacterial strains selected from the group consisting of:

[0061] Weissella viridis LB10G, deposited as DSM 32906;

[0062] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0063] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0064] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0065] Enterococcus faecium LB276R, deposited as DSM 32997;

[0066] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0067] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0068] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0069] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0070] In the present context, the term "genetic homology" relates to the deviation of the genetic sequence of a bacterial strain relative to a deposited bacterial strain.

[0071] In one embodiment of the present invention, the genetic homology may be at least 96%; such as at least 97%; such as at least 98%; such as at least 99%; such as at least 99.5%; such as at least 99.8%; such as at least 99.9%; such as 100% (identical) to a bacterial strain selected from the group consisting of:

[0072] Weissella viridis LB10G, deposited as DSM 32906;

[0073] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0074] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0075] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0076] Enterococcus faecium LB276R, deposited as DSM 32997;

[0077] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0078] - Leuconostoc mesenteroides LB341R;

[0079] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0080] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0081] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0082] In another embodiment of the present invention, the genetic homology may be at least 96%; such as at least 97%; such as at least 98%; such as at least 99%; such as at least 99.5%; such as at least 99.8%; such as at least 99.9%; such as 100% (identical) to a bacterial strain selected from the group consisting of:

[0083] Weissella viridis LB10G, deposited as DSM 32906;

[0084] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0085] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0086] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0087] Enterococcus faecium LB276R, deposited as DSM 32997;

[0088] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0089] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0090] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0091] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0092] In another embodiment of the present invention, the bacterial strain may be selected from the group consisting of: - Weissella viride LB10G, deposited as DSM 32906;

[0093] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0094] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0095] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0096] Enterococcus faecium LB276R, deposited as DSM 32997;

[0097] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0098] - Leuconostoc mesenteroides LB341R;

[0099] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0100] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0101] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0102] In yet another embodiment of the present invention, the bacterial strain may be selected from the group consisting of: - Weissella viridis LB10G, deposited as DSM 32906;

[0103] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0104] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0105] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0106] Enterococcus faecium LB276R, deposited as DSM 32997;

[0107] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0108] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0109] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0110] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0111] The effect of the bacterial strains (and / or compositions) according to the invention on pathogenic microorganisms is significant.

[0112] In one embodiment of the invention, the growth of S. aureus, such as methicillin-resistant S. aureus (MRSA), in co-culture may be reduced by at least 20%; such as by at least 30%; such as by at least 40%; such as by at least 50%, such as by at least 60%.

[0113] In a preferred embodiment, the bacterial strain according to the present invention may be an isolated bacterial strain.

[0114] The present invention discloses microorganisms according to the present invention, which are unified by their functional relationships with each other, thereby having common properties and / or effects, namely, they inhibit the growth of pathogenic microorganisms, such as Staphylococcus aureus, and / or reduce the colonization level of pathogenic microorganisms associated with skin diseases, such as Staphylococcus. These lactic acid bacteria particularly include the following microorganisms from the group of newly isolated microorganisms deposited in the German Collection for Microorganisms and Cell Cultures, or their analogs, fragments, lysates, derivatives, mutants, or combinations thereof:

[0115] Weissella viridis LB10G, deposited as DSM 32906;

[0116] - Lactobacillus paracasei LB113R, deposited as DSM 32907;

[0117] - Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0118] - Lactobacillus paracasei LB116R, deposited as DSM 32908;

[0119] Enterococcus faecium LB276R, deposited as DSM 32997;

[0120] - Lactobacillus plantarum LB316R, deposited as DSM 33091;

[0121] - Leuconostoc mesenteroides LB349R, deposited as DSM 33093;

[0122] - Lactobacillus plantarum LB356R, deposited as DSM 33094;

[0123] - Lactobacillus plantarum LB312R, deposited as DSM 33098;

[0124] The present invention includes compositions comprising at least one of these novel lactic acid bacteria, as well as compositions comprising any combination of these strains and analogs, fragments, lysates, derivatives, mutants thereof.

[0125] The inventors of the present invention provide a therapeutic composition for treating or preventing infection, comprising a therapeutically effective concentration of one or more species or strains in a pharmaceutically acceptable carrier suitable for administration to the gastrointestinal tract of a mammal and / or topical administration to the skin or mucous membranes of a mammal, wherein the probiotic strain has the ability to inhibit the growth, colonization rate and initial attachment of the pathogen to the site of infection.

[0126] Therefore, a preferred embodiment of the present invention relates to a composition comprising one or more bacterial strains according to the invention.

[0127] The concentration of bacterial strains, preferably one or more live strains and / or one or more dead strains, can be 10 3 to 10 14 In the range of 10 colony forming units (CFU); for example, in the range of 10 5 -10 13 CFU range; for example, within 10 7 -10 12 CFU range; for example, within 10 9 -10 11CFU range.

[0128] The concentration of the bacterial strain, preferably one or more dead / inactivated strains, one or more strain lysates; one or more strain metabolites may be in the range of 0.001% (w / w) to 20% (w / w).

[0129] In the context of the present invention, the bacterial strains defined herein may be provided in the composition according to the invention in the form of killed bacterial strains. The killed bacterial strains may be provided as whole dead cells or as a lysate, metabolite, derivative, analogue, fraction or extract obtained from the dead cells.

[0130] In another embodiment of the present invention, the one or more bacterial strains may be provided as one or more live strains, one or more dead or inactivated strains, one or more strain lysates, one or more strain metabolites, or a combination thereof.

[0131] In the composition, the bacterial strain according to the present invention can be provided as one or more live strains, one or more dead or inactivated strains, one or more strain lysates; one or more strain metabolites; one or more analogs, one or more fragments, one or more derivatives, one or more mutants or a combination thereof, wherein the lysate; one or more strain metabolites; one or more analogs, one or more fragments, one or more derivatives, one or more mutants or a combination thereof (as obtained from the bacterial strain according to the present invention) can treat, alleviate, inhibit, prevent and / or prevent the growth of at least one pathogenic microorganism (e.g., MRSA).

[0132] In one embodiment of the present invention, wherein the composition can be a topical composition, an oral composition or a rectal composition, preferably the composition is a topical composition.

[0133] The composition according to the present invention may preferably comprise a pharmaceutically or cosmetically acceptable carrier or excipient.In one embodiment of the present invention, the composition may be provided in solid form, liquid form, viscous form, emulsion or as a dried form.

[0134] The oral composition may preferably be formulated as a paste, soft gelatin capsule, hard gelatin capsule, powder, talc, granules, beads, pastilles, effervescent tablets, lozenges, buccal tablets, chewable tablets, sublingual tablets, oils, liquids, solutions, tinctures, emulsions, juices, concentrates, syrups, sprays, mists, drinkable ampoules, gels, tablets, coated pills or as a food or feed product or a beverage.

[0135] In one embodiment of the present invention, the composition may be a topical composition for application to human or animal skin. Compositions for topical application may preferably be formulated as a paste; a talc; a lotion; a custard; a foam; a cream; or an ointment.

[0136] In one embodiment of the present invention, the topical composition may be a powder composition comprising hydrated magnesium silicate (talc) and at least one bacterial strain of the present invention.

[0137] In a further embodiment according to the present invention, a powder composition for topical application comprises hydrated magnesium silicate, at least one carbohydrate and at least one bacterial strain according to the present invention.

[0138] In a preferred embodiment, the topical composition may be formulated as a lotion; a custard; a foam; a cream; or an ointment, oil, or emulsion.

[0139] In a preferred embodiment, the treatment is a combination treatment of a topical composition comprising the bacterial strain of the invention and an oral composition.

[0140] In addition to the bacterial strain according to the present invention, the composition may also contain other probiotics, prebiotics, antimicrobials, antibiotics or other active antimicrobial substances and / or preferably may also contain one or more substances selected from the group consisting of antioxidants, vitamins, coenzymes, fatty acids, amino acids and cofactors.

[0141] In another embodiment of the present invention, the bacterial strain according to the present invention may be combined with:

[0142] - Therapeutically effective doses of antibiotics. As combination therapy or after antibiotic treatment;

[0143] - Therapeutic concentrations of antibiotics, including but not limited to: fusidic acid; vancomycin; gentamicin; oxacillin; tetracycline; nitrofurantoin; chloramphenicol; clindamycin; trimethoprim-sulfamethoxazole, members of the cephalosporin antibiotic family (e.g., cefaclor, cefdroxil, cefixime, cefprozil, ceftriaxone, cefuroxime, cephalexin, loracarbef, etc.); members of the penicillin antibiotic family (e.g., ampicillin, amoxicillin / clavulanate, bacancillin, cloxacillin, penicillin VK, etc.); members of the fluoroquinolone antibiotic class (e.g., ciprofloxacin, grepafloxacin, levofloxacin, lomefloxacin, norfloxacin, ofloxacin, sparfloxacin, trovafloxacin, etc.); or members of the macrolide antibiotic family (e.g., azithromycin, erythromycin, etc.);

[0144] - a therapeutically effective dose of an anti-inflammatory drug as an adjunct therapy or following treatment; and / or

[0145] - Therapeutic concentrations of anti-inflammatory drugs.

[0146] In one embodiment of the present invention, the composition can be a medicament, a veterinary product, a food, a food supplement, or a food supplement composition. The composition (preferably for oral administration) may preferably contain one or more thickeners, and / or one or more sweeteners and / or one or more artificial sweeteners, wherein the thickener is preferably selected from the group consisting of cellulose ethers, polysaccharides selected from the group consisting of xanthan gum, gelatin, highly dispersed silica, starch, carrageenan, alginates, tragacanth gum, agar, gum arabic, pectin, and polyvinyl esters, and the sweetener is selected from the group consisting of glucose, fructose, sucrose, glucose syrup, sorbitol, mannitol, xylitol, maltitol, steviol glycosides, saccharin, sodium cyclamate, acesulfame K, and / or aspartame.

[0147] Preferred foods and nutritional supplements in the sense of the present invention may include effervescent tablets, vitamin tablets, dietary supplements, mineral tablets, trace element tablets, beverage powders, beverages, juices, milk drinks, yogurt, mineral water, non-carbonated water, filled gummies, chewable tablets, juices or syrups, coated pills and lozenges, and aerosols.

[0148] In addition, the composition may comprise builders, enzymes, electrolytes, pH adjusters, thickeners, prebiotics, optical brighteners, graying inhibitors, dye transfer inhibitors, foam regulators and / or colorants.

[0149] There is no disclosure in the prior art of the use of probiotic strains for the prevention or treatment of infections with pathogenic bacteria such as Staphylococcus spp. (eg MRSA) infections in skin diseases.

[0150] It was completely surprising that a group of lactic acid bacteria could be identified that share these same advantageous properties. No bacteria, and in particular no lactic acid bacteria, combine these properties: treating; alleviating; inhibiting; preventing; and / or preventing the growth of pathogenic microorganisms, while also being non-pathogenic and not causing any damage or impact on the skin or microbiome.

[0151] It should be understood that, hereinafter, preferred embodiments related to one broad aspect of the present invention are equally applicable to each of the other broad aspects of the present invention described above. It will be further understood that, unless the context indicates otherwise, the preferred embodiments described below may be combined. When used in this article, the term topical includes preparations that refer to body surfaces (e.g., skin or mucous membranes). Mucous membranes that may be mentioned in this regard include the vagina, penis, urethra, bladder, anus, oral cavity (including the mucous membranes of the cheeks, soft palate, sublingual surface, and floor of the mouth), nose, throat (including the mucous membranes of the pharynx, larynx, trachea, and esophagus), bronchi, lungs, eyes, and ears.

[0152] One embodiment of the present invention relates to a composition according to the present invention for use in treating, alleviating, inhibiting, preventing and / or preventing the growth of pathogenic microorganisms.

[0153] More preferably, the present invention may provide a composition as defined herein for use in treating, alleviating, inhibiting or preventing one or more pathogenic bacterial infections in a mammal.

[0154] More preferably, the present invention may provide a composition as defined herein for use in preventing the growth of pathogenic microorganisms.

[0155] The bacterial infection may preferably be a Staphylococcus infection in a mammal.

[0156] The staphylococcal infection may preferably be a MRSA infection.

[0157] One embodiment of the present invention relates to a composition as defined herein for use in the treatment, alleviation, inhibition and / or prevention of a disease caused by a Staphylococcal infection, such as dermatitis, atopic dermatitis, eczema, carbuncle, cellulitis, rosacea, psoriasis, diaper rash, impetigo or a wound.

[0158] In another embodiment of the present invention, a composition comprises at least one bacterial strain (or a lysate, metabolite, derivative, analog, fraction or extract thereof) for treating skin infections caused by Staphylococcus aureus, including skin infections associated with atopic dermatitis, eczema, impetigo, burns or diaper rash.

[0159] Burn patients often take antibiotics to reduce the incidence of opportunistic infections. Staphylococci are often associated with severe burn infections. Therefore, ointments, lotions, talc, gels, etc. can be combined with beneficial compositions or lysates, metabolites, derivatives, analogs, fractions, or extracts obtained from the bacterial strains of the present invention to effectively inhibit skin pathogens and prevent their growth on the patient's skin.

[0160] Another example is the use of antibiotics in livestock. The compositions disclosed herein can be applied to the skin or mucous membranes of mammals to reduce pathogen colonization.

[0161] One embodiment of a preferred invention relates to a composition for use as a prophylactic or medical treatment of Staphylococcal infections.

[0162] The microorganisms can advantageously be present in the composition in a live or killed / dead form. The bacterial strains can be provided in an encapsulated, microencapsulated, spray-dried and / or lyophilized form. In addition, the bacterial strains can be provided in the form of cell lysates, metabolites, derivatives, analogs, fractions or extracts.

[0163] In one embodiment of the present invention, the bacterial strain may be present in the composition in an amount of 0.001 wt% to 20 wt%, preferably 0.005 wt% to 10 wt%, particularly preferably 0.01 wt% to 5 wt%.

[0164] A preferred embodiment of the present invention involves administering approximately 1 x 10 3 to 1x10 14 CFU of viable bacteria, more preferably about 1x10 4 to 1x10 10 , most preferably about 5x10 per day 4 to 1x10 9 When the condition to be treated involves an antibiotic-resistant pathogen and the patient is an adult, a typical dose is about 1 x 10 CFU per day. 2 to 1x10 14 CFU of live bacteria, preferably about 1x10 8 to 1x10 10 , more preferably about 2.5 x 10 8 to 1x10 10 If the patient is an infant over 6 months old, the dose is usually 1x10 CFU per day. 6 to 1x10 9 CFU of viable bacteria.

[0165] In another aspect of the present invention, the antibiotic-resistant bacteria are resistant to at least one of the following antibiotics: fusidic acid, vancomycin, metronidazole, methicillin and / or fidaxomicin.

[0166] The present invention relates to novel bacterial strains and the general references in the claims are to living cells, dead / killed cells and lysates, metabolites, derivatives, analogs, fractions or extracts thereof and compositions comprising these living cells, dead / killed cells and lysates, metabolites, derivatives, analogs, fractions or extracts thereof.

[0167] The compositions of the present invention may be suitable for treating, alleviating, inhibiting, preventing and / or preventing the growth of pathogenic microorganisms (e.g., MRSA) in infants, toddlers, children, healthy individuals, the elderly, immunosuppressed individuals, individuals with isolated or recurrent Staphylococcus aureus infections, and / or individuals with bacterial infections that are resistant to antibiotics.

[0168] In one embodiment of the present invention, the composition according to the present invention may be suitable for treating, alleviating, inhibiting, preventing and / or preventing the growth of pathogenic microorganisms (such as MRSA) in animals (including pets and livestock).

[0169] Therefore, the composition of the present invention can be used to prepare a medicament useful for treating or preventing the growth of Staphylococci. In one embodiment of the present invention, the composition can be used for treatment or prevention, for example in combination with probiotics and / or prebiotic compositions.

[0170] The combination of a composition according to the present invention and a probiotic bacterial strain provides a combined composition capable of inhibiting the growth of Staphylococcus aureus in co-culture by reducing the growth of Staphylococcus aureus by at least 50% compared to growth in the absence of the bacterial strain, wherein growth is measured as colony forming units of Staphylococcus aureus in the stationary growth phase.

[0171] In one embodiment of the invention the bacterial strain defined herein may be the only bacteria present in the composition.A composition comprising only the bacterial strain as defined herein shows at least a 50% reduction in the growth of S. aureus compared to growth in the absence of the bacterial strain.

[0172] A "reduction" in growth can be "statistically significant" and can include a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% reduction compared to a growth period in the absence of the bacterial strain of the invention.

[0173] In one embodiment of the invention, growth inhibition may be determined as a reduction in growth of at least 25%. Preferably, growth inhibition is determined as a reduction in growth of at least 50%. Even more preferably, growth inhibition is determined as a reduction in growth of at least 90%.

[0174] A "reduction" in the number of microorganisms can be "statistically significant" and can include a 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, 99.9% or 100% reduction compared to the number of CFU / ml in the absence of the bacterial strain of the invention.

[0175] The number of microorganisms was measured as colony forming units (CFU / ml).

[0176] The microorganisms according to the invention may preferably be present in isolated or purified form, wherein the term "isolated" particularly means that the lactic acid bacteria are derived from their culture medium, including for example their natural culture medium. The term "purified" is not limited to absolute purity.

[0177] In one embodiment of the invention, the probiotic strains may be used as live isolated microorganisms in a stable form. Suitable methods for stabilization are known to those skilled in the art and include freeze drying or lyophilization involving different cryoprotectants.

[0178] In another embodiment of the present invention, the strain can be used as a live isolated strain.

[0179] Preferably, the strain can be used as a live isolated stable strain. Even more preferably, the strain can be used as a live isolated strain stabilized by lyophilization. Even more preferably, the strain can be used as a live isolated strain stabilized by lyophilization and containing a cryoprotectant.

[0180] The present invention relates to living and / or dead (killed) bacterial strains, both forms of which are included within the scope of the present invention.

[0181] Suitable killing methods (e.g., biological, chemical or physical killing methods) are well known to those skilled in the art. However, in the present case, the bacterial strains can also be used in a lyophilized form. The killed microorganisms can include the fermentation broth and any metabolites present.

[0182] The term "killed" or "dead" relates to inactivated lactic acid bacteria that are unable to undergo cell division and do not have any metabolic activity. Dead or killed lactic acid bacteria may have intact or disrupted cell membranes.

[0183] "Lysates", "derivatives", "analogs", "fractions" or "extracts" can be obtained from dead or killed lactic acid bacteria. These lysates, fractions, derivatives, analogs and extracts preferably have the property of reducing the transfer of pathogenic microorganisms between the surface of a first subject and the surface of a second subject, wherein "lysate" and the term "extract" specifically refer to a solution or suspension of microbial cells in an aqueous medium according to the present invention and contain, for example, macromolecules (such as DNA, RNA, proteins, peptides, lipids, carbohydrates, etc.) and cell debris. The lysate preferably includes cell walls or cell wall components, including binding receptors. Methods for producing lysates are well known to those skilled in the art and include, for example, the use of a "French press" or enzymatic lysis, a ball mill with glass or iron beads. Cells can be disrupted by enzymatic, physical or chemical methods. Examples of enzymatic cell lysis include individual enzymes and enzyme mixtures, such as proteases, proteinase K, lipases, glycosidases; chemical lysis can be induced by ionophores, detergents (such as SDS), acids or bases; physical methods can also be implemented by high pressure, such as using a French press, osmotic pressure, temperature changes or alternating hot and cold. Furthermore, it is of course possible to combine chemical, physical and enzymatic methods.

[0184] In a preferred embodiment, the composition and / or bacterial strain according to the present invention is suitable for treating, alleviating, inhibiting or preventing diseases associated with infection by pathogenic microorganisms in mammals.

[0185] In one embodiment, the disease may be selected from the group of skin diseases susceptible to staphylococcal infection, including psoriasis, atopic dermatitis, carbuncle, cellulitis, rosacea, psoriasis, dry skin, allergies, eczema, rash, skin irritated by ultraviolet rays, skin irritated by detergents (including irritation caused by enzymes and sodium lauryl sulfate used in detergents), thinning of the skin (e.g., the skin of the elderly and children).

[0186] The present invention also relates to a method for reducing the number of Staphylococci on the skin of a patient suffering from atopic dermatitis.

[0187] In one embodiment of the present invention, a composition comprising at least one bacterial strain can be used to control the number of Staphylococcus aureus on the skin of a patient suffering from an inflammatory skin disorder to a level at which the Staphylococcus aureus does not cause a skin infection.

[0188] In yet another embodiment of the present invention, a composition comprising at least one bacterial strain can be used to control the number of Staphylococcus aureus on the skin of a patient suffering from an inflammatory skin disorder, wherein the level of Staphylococcus aureus is less than about 10% of the skin. 5 CFU / cm 3 .

[0189] It will be clear to those skilled in the art that, herein and in all statements of ranges characterized by terms such as "about" or "approximately", it is not necessary to indicate an exact numerical range using expressions such as "approximately" or "approximately", but rather, even slight deviations upward or downward from the indicated numbers are still within the scope of the present invention. In one embodiment of the present invention, slight deviations may include deviations of 5% or less, such as 4% or less, such as 4% or less, such as 3% or less, such as 2% or less, such as 1% or less.

[0190] In one embodiment of the present invention, a biologically pure culture of one or more bacterial strains of the present invention may be provided.

[0191] In this context, the term "mammal" can include humans, primates, farm animals, sport animals, rodents, and pets. Non-limiting examples of non-human animal subjects include rodents, such as mice, rats, hamsters, and guinea pigs; rabbits; puppies; cats; sheep; pigs; piglets; sows; poultry; turkeys; broilers; minks; goats; cattle; horses; and non-human primates, such as apes and monkeys.

[0192] The term "effective amount" may depend on the context in which it is used. In the case of administering a composition to reduce the risk of a staphylococcal infection and / or administering a composition to reduce the severity of a staphylococcal infection in a subject and / or reduce the number of Staphylococcus aureus, an effective amount of a composition described herein is an amount sufficient to treat and / or ameliorate the staphylococcal infection, as well as to reduce the severity and / or reduce the likelihood of a staphylococcal infection and / or the transfer of Staphylococci between subjects. A reduction in the number of Staphylococcus aureus in a subject can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 99.9% reduction in the severity of the staphylococcal infection or the likelihood of becoming infected.

[0193] An effective amount can be administered as a composition, in one or more administrations.

[0194] An effective amount of the composition can be administered topically, orally, or a combination thereof. Topical administration is preferred.

[0195] The composition may be administered in more than one type of administration, for example in the mammal's feed or food and / or applied to the skin and / or as a nasal application.

[0196] In one embodiment of the invention, the composition comprises at least one bacterial strain according to the invention and a prebiotic.

[0197] "Prebiotics" are non-digestible food ingredients that promote the growth of specific microorganisms. "Synbiotics" are compositions containing at least one probiotic and at least one prebiotic. Such compositions are understood to promote the growth of beneficial bacteria (e.g., probiotics). Thus, powerful synbiotics are based on the combination of specific strains of probiotics with carefully selected prebiotics. They can provide important health benefits to mammals.

[0198] According to another aspect of the present invention, there is provided a probiotic composition comprising probiotic microorganisms and at least one further active ingredient.

[0199] Prebiotics are chemical products that induce the growth and / or activity of commensal microorganisms (e.g., bacteria and fungi) that contribute to the health of the host. Prebiotics are non-digestible carbohydrates that pass through the upper gastrointestinal tract undigested and stimulate the growth and / or activity of beneficial bacteria that colonize the large intestine or the skin microbiome.

[0200] Some oligosaccharides used as prebiotics are fructooligosaccharides (FOS), xylooligosaccharides (XOS), polydextrose, pectin, galacto-oligosaccharides (GOS) or human milk oligosaccharides (HMO). In addition, disaccharides such as lactulose or some monosaccharides such as lactose or tagatose can also be used as prebiotics.

[0201] In one embodiment of the present invention, at least one prebiotic compound may be included in the composition of the present invention. In a very broad concept, prebiotics are all compounds that can be metabolized by probiotics.

[0202] Preferably, prebiotics are indigestible or difficult to digest for mammals. Therefore, after being taken in by mammals, indigestible prebiotics can pass through the small intestine and enter the large intestine to stimulate the growth of probiotics in this compartment. Therefore, prebiotics can be used as a food source for probiotics. It is believed that prebiotics (many of which are indigestible carbohydrates) promote the growth of probiotics. Prebiotics are naturally present in, for example, cabbage, onion, whole grains, banana, garlic, honey, leek, artichoke, fortified food and beverage and dietary supplements. Prebiotics are well known in the art, and when used in the present invention, there is no particular limitation to prebiotics themselves.

[0203] However, in one embodiment, at least one prebiotic product in the composition is selected from the following compounds and compositions: non-digestible carbohydrates, β-glucans, manno-oligosaccharides, inulin, fructooligosaccharides, human milk oligosaccharides (HMOs), galacto-oligosaccharides (GOS), lactulose, lactofructooligosaccharides, galactotriose, fructooligosaccharides (FOS), cellobiose, cellodextrins, cyclodextrins, maltitol, lactitol, glycosilsucrose, betaine, vitamin E or a variant thereof (wherein the variant is selected from α, β, γ, δ tocopherol, tocotrienols and tocopherols). Optionally, manno-oligosaccharides and / or inulin may be preferred. HMOs may include lacto-N-tetraose, lacto-N-fucopentaose, lacto-N-triose, 3'-sialyllactose, lacto-N-neofucopentaose, sialic acid, L-fucose, 2-fucosyllactose, 6'-sialyllactose, lacto-N-neotetraose, and 3-fucosyllactose.

[0204] Prebiotics may also be used in the topical compositions of the present invention.

[0205] In one embodiment, at least one of the following prebiotic compounds is used in the topical composition of the present invention: lactose, β-glucan, manno-oligosaccharide, inulin, fructooligosaccharide, galacto-oligosaccharide (GOS), lactulose, lactofructooligosaccharide, galactotriose, fructooligosaccharide (FOS), cellobiose, cellodextrin, cyclodextrin, maltitol, lactitol, glucosylsucrose, betaine, vitamin E or a variant thereof (wherein the variant is selected from α, β, γ, δ tocopherol, tocotrienol and tocopherol), lacto-N-tetraose, lacto-N-fucopentaose, lacto-N-triose, 3'-sialyllactose, lacto-N-neofucopentaose, sialic acid, 2-fucosyllactose, 6'-sialyllactose, lacto-N-neotetraose and 3-fucosyllactose. Optionally, lactose and / or manno-oligosaccharide and / or inulin may be preferred.

[0206] D- and L-fucose strengthen the skin's natural defenses, stimulate the epidermal immune defenses and / or prevent and / or treat autoimmune diseases of the skin. In one embodiment of the invention, the composition comprises D- or L-fucose.

[0207] In one embodiment of the present invention, the composition further comprises L-fucose at a concentration of 10 mM to 500 mM in the composition.

[0208] According to still further features in the described preferred embodiments, the composition further comprises at least one active ingredient.

[0209] In one embodiment of the invention, the composition comprises at least one bacterial strain according to the invention and at least one additional probiotic microorganism selected from the group consisting of another bacterium, a yeast or a mold.

[0210] The composition according to the present invention may comprise at least one bacterial strain in combination with at least one additional probiotic microorganism, wherein the at least one additional probiotic microorganism may be selected from, but not limited to, Bifidobacterium lactis DSM10140, Bifidobacterium lactis LKM512, Bifidobacterium lactis DSM 20451, Bifidobacterium bifidum BB-225, Bifidobacterium adolescentis BB-102, Bifidobacterium breve BB-308, Bifidobacterium longum BB-536 from Zaidanhojin Nihon Bifizusukin Senta (Japan Bifidobacterium Center), Bifidobacterium NCIMB 41675 described in EP2823822. Bifidobacterium bifidum BB-225, Bifidobacterium adolescentis BB-102, Bifidobacterium breve BB-308, Bifidobacterium lactis HN019 (Howaru) available from DuPont Nutrition Biosciences ApS, Bifidobacterium lactis DN 173 010 available from Groupe Danone, Bifidobacterium breve DN 173 010 available from Chr. Hansen A / S), Bifidobacterium lactis Bb-12 available from DuPont Nutrition Biotechnology Co., Ltd., Bifidobacterium lactis 420 available from DuPont Nutrition Biotechnology Co., Ltd., Bifidobacterium breve Bb-03, Bifidobacterium lactis BI-04, Bifidobacterium lactis Bi-07 available from DuPont Nutrition Biotechnology Co., Ltd., Bifidobacterium bifidum Bb-02, Bifidobacterium bifidum Bb-06, Bifidobacterium longum KC-1 and Bifidobacterium longum 913 (DuPont Nutrition Biotechnology Co., Ltd.), Bifidobacterium breve M-16V (Morinaga) and / or lactobacilli having a probiotic effect, and may be Any of the following strains: Lactobacillus rhamnosus LGG (Chr. Hansen), Lactobacillus acidophilus NCFM (DuPont Nutrition Biotech, Inc.), Lactobacillus bulgaricus 1260 (DuPont Nutrition Biotech, Inc.), Lactobacillus paracasei Lpc-37 (DuPont Nutrition Biotech, Inc.), Lactobacillus rhamnosus HN001 (Howaru) available from DuPont Nutrition Biotech, Inc., Streptococcus thermophilus 715 and Streptococcus thermophilus ST21 available from DuPont Nutrition Biotech, Inc., Lactobacillus paracasei subsp. paracasei CRL431 (ATCC 55544), Lactobacillus paracasei strain F-19 from Medipharm, Inc., Lactobacillus paracasei LAFTI L26 (DSM Food Specialties, The Netherlands), and Lactobacillus paracasei CRL 431 (Chr. Hansen), Lactobacillus thermophilus PTA-4797, Lactobacillus salivarius Ls-33, and Lactobacillus curva 853 (DuPont Nutrition Biotech, Inc.).Lactobacillus paracasei subsp. rhamnosus LC705, Lactobacillus DSM15527 (Bifodan), Lactobacillus DSM15526 (Bifodan) described in Finnish Patent 92498, Valio Oy, Lactobacillus rhamnosus GG (LGG) (ATCC 53103) and Lactobacillus rhamnosus LC705 (DSM 7061) described in U.S. Patent 5,032,399, propionic acid bacteria (e.g., Propionibacterium freudenreichii subsp. schizophylla) PJS (DSM 7067) described in more detail in Finnish Patent 92498, Valio Oy, Nitrosomonas D23 (A Biome), Staphylococcus hominis A9, C2, AMT2, AMT3, AMT4-C2, AMT4-G1 and / or AMT4-D12 strains (all from Matrisys Bioscience), Staphylococcus epidermidis M034, M038, A11, AMT1, AMT5-C5 and / or AMT5-G6 strains (all from Matrisys Bioscience), Lactobacillus plantarum YUN-V2.0 (BCCM LMG P-29456), Lactobacillus pentosus YUN-V1.0 (BCCN LMG P-29455), Lactobacillus rhamnosus YUN-S1.0 (BCCM LMG P-2961), and / or any combination thereof.

[0211] In one embodiment of the invention, the composition comprises at least one bacterial strain as defined herein in combination with at least one strain selected from lactic acid bacteria capable of improving the integrity of tight junctions.

[0212] In another embodiment of the invention, the composition comprises at least one bacterial strain as defined herein in combination with at least one strain selected from the group consisting of Lactobacillus rhamnosus LGG (Chr. Hansen), Lactobacillus acidophilus NCFM (DuPont), Lactobacillus salivarius Ls-33 (DuPont), Propionibacterium janaschii P63 (DuPont), Bifidobacterium lactis 420 (DuPont) and Lactobacillus acidophilus La-14 (DuPont); and / or cell lysates and / or soluble metabolites of probiotic strains.

[0213] The composition according to the present invention suitable for oral administration can be taken in the form of 1×10 6 to 1×10 14 The colony forming units (CFU) were provided.

[0214] As used herein, and as is well known in the art, "treatment" is a method for obtaining a beneficial or desired result, including a clinical result. For the purposes of this subject matter, a beneficial or desired clinical result includes, but is not limited to, alleviation or amelioration of one or more symptoms, reduction in the extent of the disease, a stable (i.e., non-worsening) state of the disease, prevention of the disease, delay or slowing of disease progression, and / or improvement or remission of the disease state.

[0215] The reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% reduction in the severity of a complication or symptom.

[0216] In one embodiment of the present invention, a method for treating mammalian skin may be provided. The method comprises administering a therapeutically effective amount of at least one bacterial strain to a subject (e.g., a mammal) in need thereof, thereby treating the skin to reduce colonization and / or carrier levels and / or infection.

[0217] In another embodiment of the invention, the skin colonization may be caused by microorganisms that are resistant to antibiotics. The skin colonization may be caused by MRSA.

[0218] The at least one bacterial strain is capable of proliferating and colonizing the gastrointestinal tract, nasal passages, or skin of a mammal.

[0219] The present invention may provide several advantages. In particular, it is desirable to provide antimicrobial treatments that do not use conventional antimicrobials, as the use of antibiotics may be adversely affected by the development of antibiotic-resistant microbial species. Thus, the present invention does not contribute to the development of antibiotic-resistant pathogen progeny.

[0220] In one embodiment of the present invention, the effect of the composition according to the invention and the method according to the invention does not involve a rinsing step to remove the biofilm from the surface before adding the at least one lactic acid bacterium.

[0221] Deposit of biological materials

[0222] The following biological materials and microorganisms have been deposited in the German Collection of Microorganisms:

[0223] Weissella viridescens LB10G, deposited as DSM 32906 on August 28, 2018;

[0224] Lactobacillus paracasei LB113R, deposited under DSM 32907 on August 28, 2018;

[0225] Lactobacillus plantarum LB244R, deposited as DSM 32996 on December 13, 2018;

[0226] Lactobacillus paracasei LB116R, deposited as DSM 32908 on August 28, 2018;

[0227] Enterococcus faecium LB276R, deposited as DSM 32997 on December 13, 2018;

[0228] Lactobacillus plantarum LB316R, deposited as DSM 33091 on April 10, 2019;

[0229] Leuconostoc mesenteriodes LB349R, deposited as DSM 33093 on April 10, 2019;

[0230] Lactobacillus plantarum LB356R, deposited as DSM 33094 on April 10, 2019;

[0231] - Lactobacillus plantarum LB312R, deposited as DSM 33098, with a deposit date of April 10, 2019.

[0232] It should be noted that embodiments and features described in the context of one aspect of the invention may also apply to other aspects of the invention.

[0233] The invention will now be described in more detail in the following non-limiting examples.

[0234] Example

[0235] Example 1: Strain screening and identification

[0236] sample

[0237] In order to identify and select bacterial strains according to the present invention, a collection of lactic acid bacteria (LAB) strains was established. Samples from different sources, such as homemade sauerkraut, kimchi, and healthy human donor samples (vagina, oral cavity, anus, skin), were collected to isolate at least 995 lactic acid bacteria. The samples were anaerobically cultured overnight or until colonies formed on Man Rogosa Sharp (MRS, Sigma-Aldrich) broth and agar at 37°C, and the samples were collected. The isolates were plated and subcultured until pure colonies were obtained. The pure colonies were stored in MRS broth containing 25% glycerol at -80°C for future use. The strains were identified using standard 16S rRNS Sanger sequencing methods.

[0238] Example 2 Co-culture assay / competition assay

[0239] Competition between bacterial strains and Staphylococcus aureus was determined according to the methods described in the following publications: Dowarah, R., et al. 2018, Selection and characterization of probiotic lactic acid bacteria and its impact on growth, nutrient digestibility, health and antioxidant status in weaned piglets. PLoS ONE, 13(3), Khare, A., & Tavazoie, S. (2015). Multifactorial Competition and Resistance in a Two-Species Bacterial System. PLoS Genetics, 11(12), 1–21.).

[0240] Staphylococcus aureus subspecies COL (CCOS461), S. aureus CC1 (Bispebjerg University Hospital, Clausen et al. (2017) Br. J. Dermatol. 177:1394-1400. Doi 10.1111 / bjd.15470), and S. aureus US300 (ATCC BAA-1717) were used as test organisms. S. aureus was cultured in brain heart infusion (BHI) broth. S. aureus CC1 is a clonal type particularly associated with atopic dermatitis and is associated with FLG mutations in patients with atopic dermatitis.

[0241] The cell density of overnight cultures of Staphylococcus aureus and bacterial strain isolates was adjusted to an optical density (OD600) of 1 at 600 nm and collected by centrifugation (6000 rpm for two minutes). The cell pellet was washed twice in phosphate buffered saline (1×PBS) and resuspended in 1xPBS. 1 ml of each cell suspension was mixed in 50 ml of BHI broth and incubated at 37°C for 24 hours, while a single culture of each Staphylococcus aureus and each LAB was used as a control. At time 0, 2 hours, 6 hours, 10 hours and 24 hours, serial dilutions of the cell solution were plated on nutrient agar plates to count the colonies formed. MRS agar was used for LAB isolates and mannitol high salt phenol red agar (Sigma-Aldrich) was used for Staphylococcus aureus.

[0242] Lactobacillus rhamnosus LGG (Chr. Hansen) was used as a control probiotic strain in all experiments.

[0243] From a collection of 675 bacterial strains, 22 lactic acid bacteria (LAB) strains were identified as being able to outgrow all three tested S. aureus strains, determined to be able to reduce the growth of the tested strains by at least 25%. Eight bacterial strains were determined to have reduced growth by more than 90%. Lactobacillus rhamnosus LGG was unable to outgrow any of the S. aureus strains.

[0244] Example 3 Co-aggregation

[0245] Coaggregation was determined according to the known method Cisar, JO et al. (1979) "Specificity of Coaggregation Reactions between Human Oral Streptococci and Strains of Actinomyces Viscosus or Actinomyces Naeslundii." Infection and Immunity 24(3):742–52. Inocula of all bacterial strains were grown in MRS broth; Staphylococcus aureus strains were grown overnight in BHI broth at 37°C. Overnight cell samples were collected by centrifugation (6000 rpm for 2 minutes), and the supernatant was removed from the pellet. The pellet was washed twice in 1× PBS buffer.

[0246] The cell pellet was resuspended in 1×PBS and 500 μl of Staphylococcus aureus and bacterial strains were aliquoted into 24-well plates. The plates were incubated on a shaker (200 rpm). Self-aggregation and co-aggregation formation were visually observed after 1 hour, 2 hours, 3 hours, and 24 hours.

[0247] As a control for self-aggregation, samples of S. aureus and bacterial strains were mixed with 750 ul of PBS buffer to a final volume of 1500 ul in each well.

[0248] The plates were incubated on a shaker at approximately 200 rpm for 24 hours. Coaggregate formation was observed after 1 hour, 2 hours, 3 hours, and 24 hours.

[0249] The formation of coaggregates was scored visually using the following scale from 1-5:

[0250] 1: No aggregation

[0251] 2: Visible initial aggregation

[0252] 3: Aggregate formation <0.5mm

[0253] 4: Aggregate formation >0.5mm and <1mm

[0254] 5: Aggregate formation >1mm

[0255] Table 1: Coaggregation was measured using a visual assessment from 1 to 5 (see scale above). Data are shown for 1 hour and 24 hours of incubation.

[0256]

[0257] Example 4: Spot on lawn assay

[0258] Growth inhibition and antimicrobial metabolites were tested using lawn spot assays using the methods described in Zhang P. et al. (2015) Interstrain interactions between bacteria isolated from vacuum-packaged refrigerated beef. Appl Environ Microbiol 81:2753–2761. doi:10.1128 / AEM.03933-14 and Arena, MP et al. (2016) Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Pathogenic Microorganisms. Frontiers in Microbiology 7(APR):1–10. https: / / doi.org / 10.3389 / fmicb.2016.00464.

[0259] The bacterial strains from Example 1 were cultured from the stored samples in 2 mL of MRS broth in a 24-well plate. The S. aureus test strain was cultured in approximately 200 mL of BHI broth in an Erlenmeyer flask. The LAB isolates and S. aureus solutions were grown overnight at 37°C. The cell density of the S. aureus overnight culture was adjusted to an optical density at 600 nm (OD600) of 1 in BHI broth and then diluted to a 10^-2 dilution in PBS buffer. 200 microliters of the cell suspension were plated on BHI agar plates. The plates with the S. aureus lawns were placed in sterile air to dry for approximately 10-20 minutes. Three replicates of 20 μL of LAB isolates were found on the S. aureus lawns. The plates were left to dry and then incubated aerobically at 37°C overnight. The plates were photographed and the inhibition zones were measured in millimeters as the clearance areas around the spots. Observed growth inhibition means that the bacterial strain was able to outgrow the Staphylococcus strain in the spotted area and is indicated as (+) in Table 2. If the Staphylococcus strain was able to outgrow the spotted bacterial strain, then no growth inhibition was detected and is indicated as (-) in Table 2.

[0260] Five LAB strains were identified as exhibiting significant growth inhibition against Staphylococcus aureus, with zones of inhibition exceeding 1 mm around the spots. Lactobacillus rhamnosus LGG (Chr. Hansen) was used as a control commercial probiotic strain. Lactobacillus rhamnosus was unable to inhibit the growth of any of the Staphylococcus test strains, nor did Lactobacillus rhamnosus LGG produce any zones of clearance.

[0261] Table 2: Growth inhibition and antimicrobial metabolites. The inhibition zone was determined as the mean of three measurements.

[0262]

[0263]

[0264] A significant inhibitory effect was identified on CC1, a clonal complex of S. aureus that is particularly associated with S. aureus infection in atopic dermatitis (Clausen et al. (2017) Br. J. Dermatol. 177:1394-1400. Doi 10.1111 / bjd.15470).

[0265] Lactobacillus acidophilus NCFM, Lactobacillus salivarius Ls-33, Bifidobacterium lactis 420, Lactobacillus acidophilus La-14, and Propionibacterium jensenii P63 (all commercially available from DuPont) have been linked to improving conditions associated with tight junction function, such as atopic dermatitis, by improving the skin's barrier function. However, none of these commercial probiotic strains inhibited CC1 growth in a lawn spot test and therefore could not prevent or treat staphylococcal infections.

Claims

1. A bacterial strain having at least 95% genetic homology, for example at least 96%; such as at least 97%; for example at least 98% with one or more bacterial strains selected from the group consisting of: such as at least 99%; for example at least 99.5%; such as at least 99.8%; for example at least 99.9%; Such as 100% (same): - Lactobacillus plantarum LB244R, deposited as DSM 32996; - Lactobacillus plantarum LB356R, deposited as DSM 33094; - Lactobacillus plantarum LB316R, deposited as DSM 33091; - Lactobacillus plantarum LB312R, deposited as DSM 33098; - Lactobacillus paracasei LB113R, deposited as DSM 32907; - Lactobacillus paracasei LB116R, deposited as DSM 32908; Enterococcus faecium LB276R, deposited as DSM 32997; - Leuconostoc mesenteroides LB349R, deposited as DSM 33093; Weissella viridis LB10G, deposited as DSM 32906.

2. A bacterial strain capable of coaggregating with pathogenic microorganisms, wherein: The bacterial strain has at least 95% genetic homology, e.g. at least 96%; such as at least 97%; e.g. at least 98%; such as at least 99%; e.g. at least 99.5%; such as at least 99.8%; e.g. at least 99.9%; such as 100% (identical) to a bacterial strain selected from: - Lactobacillus plantarum LB244R, deposited as DSM 32996.

3. A bacterial strain, wherein The bacterial strain is selected from: - Lactobacillus plantarum LB244R, deposited as DSM 32996.

4. The bacterial strain according to any one of claims 1 to 3, wherein In the co-culture, the growth of S. aureus, such as methicillin-resistant S. aureus (MRSA), is reduced by at least 20%; such as by at least 30%, for example by at least 40%; such as by at least 50%, for example by at least 60%.

5. The bacterial strain according to any one of the preceding claims, wherein The bacterial strain is an isolated bacterial strain.

6. A composition comprising one or more bacterial strains according to any one of claims 1 to 5.

7. The composition according to claim 6, wherein The one or more bacterial strains are provided in the form of one or more live strains, one or more dead or inactivated strains, one or more strain lysates, one or more strain metabolites, or a combination thereof.

8. The composition according to any one of claims 6 or 7, wherein The composition is formulated as a emulsion, oil, gel, paste, powder, talc, lotion, custard, foam, cream, gel, ointment, suspension, mist or liquid.

9. The composition according to any one of claims 6 to 8, wherein The composition is a topical composition, an oral composition or a rectal composition, preferably the composition is a topical composition.

10. The composition according to any one of claims 6 to 9, wherein The concentration of the bacterial strain, preferably one or more dead / inactivated strains, one or more strain lysates, or one or more strain metabolites is 0.001% (w / w) to 20% (w / w).

Images