Strains, composition and method of use

Abstract

To provide microorganisms, compositions, and methods for treating pathogenic bacterial infections in a subject such as a mammal.SOLUTION: The present invention provides a bacterial strain selected from Lactobacillus plantarum, where the Lactobacillus plantarum is Lactobacillus plantarum LB356R deposited as DSM 33094, and a composition comprising the bacterial strain.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to novel bacterial stains. In particular, the present invention relates to novel bacterial strains for use in treating, mitigating, inhibiting, preventing, and / or preventing the growth of pathogenic microorganisms. [Background technology]

[0002] The gram-positive bacterium Staphylococcus aureus is one of the most frequently encountered human pathogens. S. aureus is particularly common in the human nasal cavity, where it is present either intermittently or persistently in approximately 50% of the human population. Although S. aureus colonization is mostly asymptomatic, the pathogen can cause infection if the skin's protective barrier function is compromised. Therefore, staphylococcal infections are often associated with skin disorders such as dermatitis, eczema, carbuncles, cellulitis, rosacea, psoriasis, diaper rash, impetigo, and wounds.

[0003] An example of a skin infection is atopic dermatitis, which is a chronic or chronically relapsing inflammatory skin disease resulting from a complex interplay of environmental, immunological, genetic and pharmacological factors.

[0004] All of these genetic and environmental factors contribute to the following characteristics: 1: Abnormal microbial colonization by a pathogenic organism such as Staphylococcus aureus (compared to Staphylococcus epidermidis in normal individuals) subsequently increases the patient's susceptibility to skin infections; additionally, Staphylococcus aureus produces enterotoxins and induces the production of enterotoxin-specific IgE, which results in greater T cell recruitment; 2. Involved in the early phase of the disease, which results in increased immunoglobulin E (IgE) production; 3: Skin barrier dysfunction or dry skin due to abnormal lipid metabolism and / or epidermal structural protein formation, and 4: Along with the psychosomatic effects of the patient due to an imbalance in the autonomic nervous system, there is then an increase in the production of mediators from various inflammatory cells.

[0005] Atopic dermatitis is typically treated with topical moisturizers / emollients or corticosteroids as first-line therapies, followed by topical calcineurin inhibitors as a second-line treatment for patients not controlled on first-line treatment.

[0006] Currently, there is no topical treatment for patients with mild to moderate atopic dermatitis that is not accompanied by serious side effects such as local thinning of the skin at the application site, exacerbation or risk of acne in the patient, and a burning sensation.

[0007] Unmet needs for atopic dermatitis patients remain. Topical treatments still cause patients distress as itching is a well-known side effect. Scratching simply worsens the disease symptoms, leading to lichenification, excoriation, and breakdown of the skin barrier, creating a vicious cycle for disease sufferers and increasing the risk of skin infections.

[0008] The most challenging unmet need yet to be addressed is the prevention and treatment of skin infections caused by Staphylococcus aureus, especially staphylococci that are resistant to antibiotics, which evade additional antibiotic resistance.

[0009] Furthermore, many different pathogenic microorganisms have been discovered and can include bacterial, viral, fungal, parasitic, and algal microorganisms, with the gram-positive bacterium Staphylococcus aureus being one of the most frequently encountered human pathogens.

[0010] Following the discovery of penicillin in 1928 and its mass production in the early 1940s, infections caused by Staphylococcus aureus could usually be treated without any major complications. However, clinicians soon observed the emergence of penicillin-resistant strains of S. aureus, primarily due to the bacterial expression of β-lactamases, enzymes that disrupt the β-lactam ring structure of the penicillin and cephalosporin classes of antibiotics (β-lactam antibiotics), thereby destroying their antibacterial activity. Methicillin, a new penicillin analogue resistant to β-lactamases, was introduced in 1959 and was initially effective against penicillin-resistant S. aureus strains. However, this success was short-lived, as the first methicillin-resistant S. aureus (MRSA) strains were identified in the laboratory by 1961, and cases of MRSA were first observed in clinics in 1968.

[0011] MRSA infections primarily occur in hospital settings, and MRSA is currently one of the most common nosocomial pathogens and is therefore a major cause of various hospital-acquired infections (HAIs). Since 1990, a new type of MRSA known as community-associated MRSA (CA-MRSA) has emerged. CA-MRSA is not only genetically distinct from healthcare-associated MRSA (HA-MRSA) strains, but can also exhibit different virulence and antibiotic resistance patterns. In recent years, hybridization between different CA-MRSA and HA-MRSA strains has been observed, which often makes it difficult to determine the origin of the infecting MRSA strain.

[0012] HA-MRSA infections are associated with major clinical complications and occur frequently in patients on mechanical ventilation and in patients who have surgical site wounds or surgical implants or who require surgical wound drainage. Furthermore, HA-MRSA infections often lead to nosocomial pneumonia or bacteremia, which are associated with high morbidity and mortality.

[0013] Staphylococcus aureus not only infects humans but also other mammals, so infected livestock and pets are an additional source of transmission. Livestock-associated MRSA (LA-MRSA) has mostly been identified in swine herds, with colonization rates varying dramatically from 10 to 80%, although LA-MRSA has also been found in ruminants and poultry.

[0014] LA-MRSA is becoming an increasing problem for the livestock industry. Objects such as stables, livestock sheds, animals, farmers, farmers' families, stable owners and visitors, as well as slaughterhouses and animal transport vehicles are contaminated with LA-MRSA. These objects can carry LA-MRSA, which may be the reason for LA-MRSA to be transferred between subjects, thus spreading LA-MRSA among healthy carriers, and may also pose an additional risk of MRSA being transmitted through carriers to hospitals or nursing homes, and to patients suffering from inflammatory skin diseases such as eczema and atopic dermatitis, carbuncles, cellulitis, rosacea, psoriasis, wounds and burns, or to humans, including children and infants, who may suffer from diaper rash and impetigo caused by MRSA.

[0015] Therefore, a treatment for pathogenic microorganisms such as MRSA would be advantageous, and in particular a more efficient and / or reliable treatment that does 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 for humans, including children and young children, resulting in diaper rash and impetigo, and that does not induce further resistance, e.g., antibiotic resistance, in pathogenic microorganisms. Summary of the Invention [Problem to be solved by the invention]

[0016] Accordingly, an object of the present invention relates to microorganisms, compositions and methods for treating pathogenic bacterial infections in subjects, such as mammals. [Means for solving the problem]

[0017] In particular, it is an object of the present invention to provide microorganisms, compositions and methods for treating pathogenic bacterial infections in subjects, such as mammals, that overcome the above-mentioned problems of the prior art associated with skin infections, skin diseases, morbidity and mortality resulting from infection and antibiotic resistance.

[0018] Thus, one aspect of the present invention is -Weissella viridescens LB10G, deposited as DSM32906; Lactobacillus paracasei LB113R, deposited as DSM32907; Lactobacillus plantarum LB244R, deposited under the designation DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB341R; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; The present invention relates to a bacterial strain having at least 95% genetic homology to one or more of the bacterial strains selected from the group consisting of:

[0019] Therefore, another aspect of the present invention is -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; The present invention relates to a bacterial strain having at least 95% genetic homology to one or more of the bacterial strains selected from the group consisting of:

[0020] Another aspect of the present invention relates to a composition comprising one or more of the bacterial strains according to the present invention.

[0021] Yet another aspect 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.

[0022] Detailed Description of the Invention The present invention relates to probiotic bacteria and compositions for use in preventing or treating infectious diseases caused by pathogenic microorganisms, such as staphylococcal infections. The present invention also relates to compositions and new microbial strains that can inhibit the growth of pathogenic microorganisms, such as MRSA.

[0023] MRSA can be spread from one subject to another through direct contact with a carrier, such as an infected surface or another infected person, such as by sharing a personal item that has come into contact with infected skin, or by touching a contaminated object, including skin, nasal passages, animals, surfaces, or objects.

[0024] Despite improvements in treatment, invasive MRSA infections, especially those acquired in community or livestock environments, remain a problem, resulting in substantial discomfort for patients as well as a significant financial burden for health care systems. Similarly, in the European Union, MRSA accounts for 44% of all healthcare-associated infections (HAIs), 22% of attributable excess deaths, and 41% of excess hospital days associated with HAIs. MRSA is a serious public health concern in Japan, which has one of the highest crude prevalence rates of MRSA among various Staphylococcus aureus strains in the world.

[0025] The present invention provides new microbial strains and new compositions that can inhibit the growth of Staphylococcus aureus without contributing to the further development of antibiotic resistance.

[0026] Therefore, a preferred embodiment of the present invention comprises: -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB341R; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; The present invention relates to a bacterial strain having at least 95% genetic homology to one or more of the bacterial strains selected from the group consisting of:

[0027] Therefore, another preferred embodiment of the present invention comprises: -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; The present invention relates to a bacterial strain having at least 95% genetic homology to one or more of the bacterial strains selected from the group consisting of:

[0028] In the present context, the term "genetic homology" relates to the deviation of the genetic sequence of the bacterial strain relative to the deposited bacterial strain.

[0029] In an embodiment of the invention, the genetic homology is -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB341R; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; at least 96%, such as at least 97%, at least 98%, such as at least 99%, at least 99.5%, such as at least 99.8%, at least 99.9%, such as 100% (identity), to one of the bacterial strains selected from the group consisting of:

[0030] In an embodiment of the invention, the genetic homology is -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; Leuconostoc mesenteriodes LB349R, deposited as DSM33093 - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; at least 96%, such as at least 97%, at least 98%, such as at least 99%, at least 99.5%, such as at least 99.8%, at least 99.9%, such as 100% (identity), to one of the bacterial strains selected from the group consisting of:

[0031] Therefore, a preferred embodiment of the present invention comprises: -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB341R; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; may be selected from the group consisting of:

[0032] Therefore, in a yet further preferred embodiment of the present invention, the bacterial strain is -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; Lactobacillus plantarum LB312R, deposited as DSM33098 ; may be selected from the group consisting of:

[0033] The effect of the bacterial strains (and / or compositions) according to the invention against pathogenic microorganisms is significant.

[0034] In one embodiment of the invention, the growth of Staphylococcus aureus, such as methicillin-resistant Staphylococcus aureus (MRSA), in co-culture can be reduced by at least 40%, such as by at least 20%, such as by at least 30%, for example by at least 50%, for example by at least 60% reduction.

[0035] In a preferred embodiment, the bacterial strain according to the present invention may be an isolated bacterial strain.

[0036] The present invention discloses that microorganisms related to each other through a functional relationship so as to form a uniform concept according to the invention share properties and / or effects, i.e., inhibit the growth of pathogenic microorganisms such as Staphylococcus aureus and / or reduce the colonization level of pathogenic microorganisms such as Staphylococcus associated with skin diseases. These lactic acid bacteria include, in particular, those from the German Collection for The following newly isolated microorganisms deposited in Microorganisms and Cell Cultures, namely: -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; and the like, or an analog, fragment, lysate, derivative, mutant or combination thereof.

[0037] The present invention encompasses compositions comprising at least one of these new lactic acid bacteria, as well as compositions comprising any combination of these strains and analogs, fragments, lysates, derivatives, and mutants thereof.

[0038] The inventors of the present invention provide a therapeutic composition for treating or preventing infectious diseases 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 mucosa of a mammal, wherein the probiotic strains have the ability to inhibit pathogen growth, colonization rate and initial attachment of the pathogen to the site of infection.

[0039] Hence, a preferred embodiment of the present invention relates to a composition comprising one or more of the bacterial strains according to the present invention.

[0040] The concentration of bacterial strains, preferably one or more viable strains and / or one or more dead strains, is 10 5 ~10 13 Colony forming units (CFU) range, e.g., 10 9 ~10 11 CFU range, etc., 10 7 ~10 12 CFU range, e.g. 10 3 ~10 14 CFU range.

[0041] The concentration of the bacterial strain, preferably one or more killed / inactivated strains, one or more strains, one or more lysates, one or more strain metabolites may be at a concentration of 0.001% (w / w) to 20% (w / w).

[0042] 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, which may be provided as whole dead cells or as a lysate, metabolite, derivative, analogue, fraction or extract obtained from the dead cells.

[0043] In further embodiments of the invention, one or more of the bacterial strains may be provided as one or more viable strains, one or more killed or inactivated strains, one or more strain lysates, one or more strain metabolites, or a combination thereof.

[0044] In the composition, the bacterial strain according to the invention may be provided as one or more viable strains, one or more killed or inactivated strains, one or more strain lysates, one or more strain metabolites, one or more analogues, one or more fragments, one or more derivatives, one or more mutants or combinations thereof, wherein the lysates, one or more strain metabolites, one or more analogues, one or more fragments, one or more derivatives, one or more mutants or combinations thereof (as obtained from the bacterial strain according to the invention) are capable of treating, mitigating, inhibiting, preventing and / or preventing the growth of at least one pathogenic microorganism, for example MRSA.

[0045] In embodiments of the present invention in which the composition may be a topical, oral or rectal composition, preferably the composition is a topical composition.

[0046] The composition according to the invention may preferably comprise a pharmaceutically or cosmetically acceptable vehicle or excipient. In one embodiment of the invention, the composition may be provided in solid, liquid, viscous, emulsion or dry form.

[0047] Compositions for oral consumption may preferably be formulated into pastes, soft gelatin capsules, hard gelatin capsules, powders, 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, gums, tablets, coated pills, or as a food or feed product or beverage.

[0048] In one embodiment of the present invention, the composition may be a topical composition for either human or animal skin. Compositions for topical application may preferably be formulated into a paste, talc, lotion, custard, foam, cream, or ointment.

[0049] In one embodiment of the present invention, the topical composition may be a powder composition comprising hydrated magnesium silicate (talc) and at least one of the bacterial strains of the present invention.

[0050] In a further embodiment according to the present invention, a powder composition for topical application may be a powder composition comprising hydrated magnesium silicate, at least one carbohydrate, and at least one of the bacterial strains of the present invention.

[0051] In preferred embodiments, the topical composition may be formulated into a lotion, custard, foam, cream, or ointment, oil, or emulsion.

[0052] In a preferred embodiment, the treatment is a combination treatment with both a topical and an oral composition comprising a bacterial strain according to the present invention.

[0053] The composition may further comprise, in addition to the bacterial strain according to the invention, other probiotics, prebiotics, antimicrobials, antibiotics or other active antimicrobial substances and / or may also preferably contain one or more of the following substances selected from antioxidants, vitamins, coenzymes, fatty acids, amino acids and cofactors.

[0054] In another embodiment of the invention, the bacterial strain according to the invention may be combined with: Therapeutically effective doses of antibiotics, either as co-treatment or following antibiotic therapy. Therapeutic concentrations of antibiotics, including but not limited to: fusidic acid, vancomycin, gentamicin, oxacillin, tetracycline, nitroflurantin, chloramphenicol, clindamycin, trimethoprim-sulfamethoxazole, members of the cephalosporin antibiotic family (e.g., cefaclor, cefadroxil, cefixime, cefprozil, ceftriaxone, cefuroxime, cephalexin, loracarbef, etc.), penicillin-based antibiotics members of the fluoroquinolone family of antibiotics (e.g., ciprofloxacin, grepafloxacin, levofloxacin, lomefloxacin, norfloxacin, ofloxacin, sparfloxacin, trovafloxacin, etc.); or members of the macrolide antibiotic family (e.g., azithromycin, erythromycin, etc.), A therapeutically effective dose of an anti-inflammatory drug, either as a co-treatment or post-therapy, and / or -Therapeutic concentrations of anti-inflammatory drugs.

[0055] In one embodiment of the present invention, the composition may be a pharmaceutical, veterinary drug, or food or nutritional supplement or nutraceutical 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, the thickeners preferably being polysaccharides selected from the group including cellulose ether, xanthan gum, gelatin, highly dispersed silicon dioxide, starch, carrageenan, alginate, tragacanth, agar, gum arabic, pectin and polyvinyl ester, and the sweeteners selected from the group including glucose, fructose, sucrose, glucose syrup, sorbitol, mannitol, xylitol, maltitol, stewia, saccharin, sodium cyclamate, acesulfame K and / or aspartame.

[0056] Preferred foods and dietary supplements in the sense of the present invention may include effervescent tablets, vitamin tablets, dietary supplements, mineral tablets, trace element tablets, drink powders, beverages, juices, milk drinks, yogurt, mineral water, non-carbonated water, bonbons, chewable tablets, juices or syrups, coated pills and lozenges, and aerosols.

[0057] Additionally, the compositions may also contain builders, enzymes, electrolytes, pH adjusters, thickeners, prebiotics, fluorescent agents, graying inhibitors, dye transfer inhibitors, foam regulators and / or colorants.

[0058] In the prior art, pathogenic Staphylococcus supp, e.g., MRSA, in skin diseases There is no disclosure of the use of probiotic strains to prevent or treat bacterial infections.

[0059] It was completely surprising to be able to identify a group of lactic acid bacteria with the same advantageous properties: bacteria, and in particular lactic acid bacteria, have not combined these properties to treat, mitigate, inhibit, prevent and / or prevent pathogenic microbial growth, while also being non-pathogenic and without causing or affecting any damage to the skin or microflora.

[0060] It will be understood that, hereinafter, preferred embodiments mentioned with respect 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 the preferred embodiments described below can be combined unless the context dictates otherwise. As used herein, the term "topical" includes reference to a formulation adapted for application to a body surface (e.g., skin or mucous membrane). Mucous membranes that may be mentioned in this regard include those of the vagina, penis, urethra, bladder, anus, mouth (including the mucous membranes of the cheeks, soft palate, underside of the tongue, and floor of the mouth), nose, throat (including the mucous membranes of the pharynx, larynx, trachea, and esophagus), bronchi, lungs, eyes, and ears.

[0061] 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.

[0062] More preferably, the present invention provides compositions as defined herein for use in treating, alleviating, suppressing or preventing one or more pathogenic bacterial infections in a mammal.

[0063] More preferably, the present invention provides compositions as defined herein for use in preventing the growth of pathogenic microorganisms.

[0064] The bacterial infection may preferably be a staphylococcal infection in a mammal.

[0065] The staphylococcal infection may preferably be an MRSA infection.

[0066] One embodiment of the present invention relates to a composition as defined herein for use in the treatment, alleviation, suppression and / or prevention of diseases resulting from a staphylococcal infection, such as dermatitis, atopic dermatitis, eczema, carbuncles, cellulitis, rosacea, psoriasis, diaper rash, impetigo or wounds.

[0067] In a further embodiment of the invention, the composition comprises at least one of the bacterial strains (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.

[0068] Burn patients are typically administered antibiotics to reduce the incidence of opportunistic infections. Staphylococcus species are often associated with severe burn infections. For this reason, salves, 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 achieve inhibition of skin pathogens and be effective in preventing the growth of pathogens on the skin of patients.

[0069] Another example is the use of antibiotics in livestock, where the compositions disclosed in the present invention can be administered to the skin or mucous membranes of mammals to reduce pathogen colonization.

[0070] A preferred invention embodiment relates to a composition for use as a prophylactic or medical treatment for staphylococcal infections.

[0071] The microorganisms can advantageously be present in the composition in a viable form or in a killed / dead form. The bacterial strains can be provided in an encapsulated, microencapsulated, spray-dried and / or lyophilized form. Furthermore, the bacterial strains can be provided in the form of cell lysates, metabolites, derivatives, analogs, fractions or extracts.

[0072] In one embodiment of the present invention, the bacterial strain may be present in the composition in an amount of 0.001% to 20% by weight, preferably 0.005% to 10% by weight, particularly preferably 0.01% to 5% by weight.

[0073] A preferred embodiment of the present invention is approximately 1 x 10 per day 3 ~1×10 14 CFU of viable bacteria, more preferably approximately 1 x 10 per day 4 ~1×10 10 , and most preferably approximately 5×10 per day 4 ~1×10 9 When the condition being treated involves an antibiotic-resistant pathogen and the patient is an adult, a typical dosage is approximately 1 x 10 per day. 2~1×10 14 CFU of viable bacteria, preferably about 1 x 10 per day 8 ~1×10 10 , and more preferably approximately 2.5×10 per day 8 ~1×10 10 If the subject being treated is an infant over 6 months of age, the dosage is typically 1 x 10 per day 6 ~1×10 9 CFU of viable bacteria.

[0074] In a further aspect of the present invention, the antibiotic-resistant bacteria is resistant to at least one of the following antibiotics: ficidic acid, vancomycin, metronidazole, methicillin and / or fidaxomicin.

[0075] The present invention relates to novel bacterial strains and general references in the claims to live cells, dead / killed cells and their lysates, metabolites, derivatives, analogues, fractions or extracts, as well as compositions comprising such live cells, dead / killed cells and their lysates, metabolites, derivatives, analogues, fractions or extracts.

[0076] The compositions 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 infants, toddlers, children, healthy individuals, the elderly, immunosuppressed individuals, individuals with single or recurrent Staphylococcus aureus infections and / or individuals with antibiotic-resistant bacterial infections.

[0077] In one embodiment of the present invention, the compositions according to the present invention may be suitable for treating, mitigating, inhibiting, preventing and / or preventing the growth of pathogenic microorganisms, such as MRSA, in animals, including pets and livestock.

[0078] The compositions of the present invention may therefore be used to prepare medicaments that are useful in treating or preventing the growth of staphylococci. In one embodiment of the present invention, the compositions may be used curatively or prophylactically, for example in combination with probiotic and / or prebiotic compositions.

[0079] The combination of the composition and probiotic strain according to the present invention provides a combined composition capable of inhibiting the growth of S. aureus in co-culture by reducing the growth of S. aureus by at least 50% compared to growth without the bacterial strain, where growth is measured as colony forming units in the stationary growth phase of S. aureus. .

[0080] In one embodiment of the invention, the bacterial strains defined herein may be the only bacteria present in the composition, and compositions comprising only the bacterial strains defined herein exhibit at least a 50% reduction in the growth of Staphylococcus aureus compared to growth without the bacterial strain.

[0081] The "reduction" in growth may be "statistically significant" compared to the growth period in the absence of the bacterial strain of the present invention and may 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.

[0082] In one embodiment of the present invention, growth inhibition may be determined as at least a 25% reduction in growth. Preferably, growth inhibition is determined as at least a 50% reduction in growth. Even more preferably, growth inhibition is determined as at least a 90% reduction in growth.

[0083] The "reduction" in the number of microorganisms may be "statistically significant" compared to the number of CFU / ml in the absence of the bacterial strain of the present invention and may 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.

[0084] The number of microorganisms is measured as colony forming units, CFU / ml.

[0085] The microorganism according to the invention may preferably be in isolated or purified form, where the term "isolated" particularly means that the lactic acid bacteria are derived from a culture medium, including, for example, the natural culture medium of the lactic acid bacteria. The term "purified" is not restricted to absolute purity.

[0086] In one embodiment of the present invention, the probiotic strains may be used as live isolated microorganisms in a stabilized form. Suitable methods for stabilization are known to those skilled in the art and include freeze drying or lyophilization involving different cryoprotectants.

[0087] In a further embodiment of the invention, the strain may be used as a live isolate.

[0088] Preferably, the strain can be used as a live isolate stabilized strain. Even more preferably, the strain can be used as a live isolate stabilized by lyophilization. Even more preferably, the strain can be used as a live isolate stabilized by lyophilization and containing a cryoprotectant.

[0089] The present invention relates to bacterial strains that are viable and / or dead (killed), and both forms may be included within the scope of the present invention.

[0090] Suitable methods for killing (e.g., biological, chemical or physical killing methods) are well known to those skilled in the art. However, in this case, the bacterial strain may also be used in a freeze-dried form. The killed form of the microorganism may include the fermentation broth and any present metabolic products.

[0091] The term "killed" or "dead" relates to inactivated lactic acid bacteria that are incapable of cell division and do not have any metabolic activity. or may have a ruptured cell membrane.

[0092] "Lysates," "derivatives," "analogs," "fractions," or "extracts" can be obtained from killed 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 object and the surface of a second object, where the terms "lysate" and "extract" refer specifically to a solution or suspension of cells of the microorganism according to the present invention in an aqueous medium, including macromolecules such as DNA, RNA, proteins, peptides, lipids, carbohydrates, and cell debris. The lysate preferably contains 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, or a ball mill using glass or iron beads. Cells can be disrupted by enzymatic, physical, or chemical methods. Examples of enzymatic cell lysis can include individual enzymes as well as enzyme cocktails, such as proteases, proteinase K, lipases, glycosidases, chemical lysis can be induced by ionophores, detergents such as SDS, acids or bases, and physical methods can also be performed using high pressure such as a French press, osmolality, temperature, or alternating heat and cold. Furthermore, chemical, physical, and enzymatic methods can of course be combined.

[0093] In a preferred embodiment, the compositions and / or bacterial strains according to the present invention are suitable for treating, alleviating, inhibiting or preventing diseases associated with pathogenic microbial infections in mammals.

[0094] In one embodiment, the disease may be selected from the group of skin diseases susceptible to staphylococcal infections, including psoriasis, atopic dermatitis, carbuncles, cellulitis, rosacea, dry skin, allergies, eczema, rashes, UV-irritated skin, detergent-irritated skin (including irritation caused by enzymes used in cleaning detergents and by sodium lauryl sulfate), thinning skin (e.g., elderly and pediatric skin).

[0095] The present invention also relates to a method for reducing the number of Staphylococcus aureus on the skin of a patient with atopic dermatitis.

[0096] In one embodiment of the present invention, a composition comprising at least one bacterial strain may be used to control the number of Staphylococcus aureus on the skin of a patient with an inflammatory skin disease to a level where the Staphylococcus aureus does not result in infection of the skin.

[0097] In a further embodiment of the present invention, a composition comprising at least one bacterial strain is used to treat a bacterial infection in which the level of Staphylococcus aureus is approximately 10 5 CFU / 1cm of skin 3 The present invention can control the number of Staphylococcus aureus on the skin of patients with inflammatory skin diseases to less than 100 mg / kg.

[0098] Herein, and in all descriptions of ranges given in the present invention characterized by terms such as "about" or "approximately," it will be apparent to one skilled in the art that the exact numerical range need not be indicated by expressions such as "about" or "approximately," but instead slight deviations above and below the stated numbers will still fall within the scope of the present invention. In one embodiment of the present invention, slight deviations may include deviations of 4% or less, e.g., deviations of 3% or less, such as deviations of 2% or less, deviations of 5% or less, e.g., deviations of 1% or less.

[0099] In one embodiment of the present invention, biologically pure cultures of one or more of the bacterial strains of the present invention may be provided.

[0100] In the present context, the term "mammal" may 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, dogs, cats, sheep, pigs, piglets, sows, poultry, turkeys, broilers, mink, goats, cattle, horses, and non-human primates, such as apes and monkeys.

[0101] The term "effective amount" may depend on the context in which it is applied. In the context of administering a composition for reducing the risk of staphylococcal infection, and / or administering a composition for reducing the severity of staphylococcal infection, and / or reducing the amount of Staphylococcus aureus in a subject, an effective amount of the composition described herein is an amount sufficient to treat and / or alleviate staphylococcal infection, and reduce the severity of staphylococcal infection and / or reduce the transmission of staphylococcus between subjects and / or the likelihood of the infection. The reduction in the amount of Staphylococcus aureus in a subject may be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 99.9% reduction in the severity of staphylococcal infection or the likelihood of infection.

[0102] An effective amount can be administered as a composition in one or more administrations.

[0103] An effective amount of the composition can be administered topically, orally, or a combination thereof, preferably topically.

[0104] The composition may be applied in two or more administrations, such as in the feed or food for the mammal, and / or may be applied to the skin and / or as a nasal application.

[0105] In one embodiment of the invention, the composition comprises at least one bacterial strain according to the invention and a prebiotic.

[0106] "Prebiotics" are indigestible food ingredients that promote the growth of specific microorganisms. "Synbiotics" are compositions that contain at least one probiotic and at least one prebiotic. Such compositions are understood to promote the growth of beneficial bacteria (e.g., probiotics). Therefore, powerful synbiotics are based on the combination of specific strains of probiotic bacteria with carefully selected prebiotics. Such synbiotics may lead to important health benefits for mammals.

[0107] According to another aspect of the present invention, there is provided a probiotic composition comprising a probiotic microorganism and at least one further active ingredient.

[0108] Prebiotics refer to chemical products that induce the growth and / or activity of commensal microorganisms (e.g., bacteria and fungi) that contribute to the health of their host. Prebiotics are non-digestible carbohydrates that pass undigested through the upper gastrointestinal tract and stimulate the growth and / or activity of beneficial bacteria that colonize the large intestine or skin microbiome.

[0109] Some oligosaccharides used as prebiotics are fructooligosaccharides (FOS), xylooligosaccharides (XOS), polydextrose, pectin, galactooligosaccharides (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.

[0110] 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 sense, prebiotics are all compounds that can be metabolized by probiotics.

[0111] Preferably, prebiotics are indigestible or difficult to digest by mammals.Therefore, after being taken in by mammals, indigestible prebiotics can pass through the small intestine and enter the large intestine, and stimulate the growth of probiotics in this compartment.Therefore, prebiotics can serve as a food source for probiotics.Prebiotics, most of which are indigestible carbohydrates, are thought to promote the growth of probiotics.Prebiotics are naturally found in, for example, cabbage, onion, whole grains, banana, garlic, honey, chives, artichoke, enriched foods and beverages, and nutritional supplements.Prebiotics are well known in the art, and there is no particular limitation to the prebiotics themselves when used in the present invention.

[0112] However, in one embodiment, at least one prebiotic product in the composition is selected from the following compounds and compositions: indigestible carbohydrates, β-glucan, mannan oligosaccharides, inulin, oligofructose, human milk oligosaccharides (HMO), galactooligosaccharides (GOS), lactulose, lactosucrose, galactotriose, fructooligosaccharides (FOS), cellobiose, cellodextrin, cyclodextrin, maltitol, lactitol, glycosyl sucrose, betaine, vitamin E or its variants (the variants are selected from alpha tocopherol, beta tocopherol, gamma tocopherol, delta tocopherol, tocotrienol and tocomonoenol).In some cases, mannan oligosaccharides and / or inulin may be preferred. HMOs can 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.

[0113] Prebiotics may also be used in the topical compositions of the present invention.

[0114] In one embodiment, at least one of the following prebiotic compounds is used in the topical compositions of the present invention: Namely, lactose, β-glucan, mannan oligosaccharides, inulin, oligofructose, galactooligosaccharides (GOS), lactulose, lactosucrose, galactotriose, fructooligosaccharides (FOS), cellobiose, cellodextrin, cyclodextrin, maltitol, lactitol, glycosylsucrose, betaine, vitamin E or a variant thereof (the variant being selected from alpha tocopherol, beta tocopherol, gamma tocopherol, delta tocopherol, tocotrienol and tocomonoenol), 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. In some cases, lactose and / or mannan oligosaccharides and / or inulin may be preferred.

[0115] D-fucose 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 present invention, the composition comprises D-fucose or L-fucose.

[0116] 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.

[0117] According to still further features in the described preferred embodiments the composition further comprises at least one active ingredient.

[0118] In one embodiment of the invention, the composition comprises at least the bacterial strain of the invention in combination with at least one further probiotic microorganism selected from the group consisting of another bacterium, a yeast or a mold.

[0119] The composition according to the invention may comprise at least one bacterial strain in combination with at least one further probiotic microorganism, wherein said at least one further probiotic microorganism may be selected from, but is not limited to: These include Bifidobacterium lactis DSM10140, Bifidobacterium lactis LKM512, Bifidobacterium lactis DSM20451, Bifidobacterium bifidum BB-225, Bifidobacterium adolescentis BB-102, Bifidobacterium breve BB-308, Bifidobacterium longum BB-536 manufactured by the Japan Bifidobacteria Center, and Bifidobacterium NCIMB41675 described in European Patent No. 2823822. Bifidobacterium bifidum BB-225, Bifidobacterium adolescentis BB-102, Bifidobacterium breve BB-308, Bifidobacterium lactis HN019 (Howaru) available from DuPont Nutrition Biosciences ApS, Bifidobacterium lactis DN173010 available from Groupe Danone, Chr.Bifidobacterium lactis Bb-12 available from Hansen A / S, Bifidobacterium lactis 420 available from DuPont Nutrition Biosciences ApS, Bifidobacterium breve Bb-03, Bifidobacterium lactis BI-04, Bifidobacterium lactis BI-07 available from DuPont Nutrition Biosciences ApS, Bifidobacterium bifidum Bb-02, Bifidobacterium bifidum Bb-06, Bifidobacterium longum KC-1 and Bifidobacterium longum 913 (DuPont Nutrition Biosciences ApS), Bifidobacterium breve M-16V (Morinaga) and / or probiotic lactobacillus, which may be any of the following strains: Lactobacillus rhamnosus LGG (Chr. Hansen), Lactobacillus acidophilus NCFM (DuPont Nutrition Biosciences ApS), Lactobacillus bulgaricus 1260 (DuPont Nutrition Biosciences ApS), Lactobacillus paracasei Lpc-37 (DuPont Nutrition Biosciences ApS), Lactobacillus rhamnosus HN001 available from DuPont Nutrition Biosciences ApS (Howaru), DuPont Nutrition Biosciences The strains tested were Streptococcus thermophilus 715 and Streptococcus thermophilus ST21 available from ApS, Lactobacillus paracasei subsp. paracasei CRL431 (ATCC 55544), and Lactobacillus paracasei F-19 strain from Medipharm, Inc. Lactobacillus paracasei LAFTI L26 (DSM Food Specialties, Oran). The strains used were Lactobacillus paracasei CRL431 (Chr. Hansen), Lactobacillus acidophilus PTA-4797, Lactobacillus salivarius Ls-33, and Lactobacillus curvatus 853 (DuPont Nutrition Biosciences ApS). Lactobacillus casei subsp. rhamnosus LC705 is described in Finnish Patent Invention No. 92498 (Valio Oy), Lactobacillus DSM15527 (Bifodan), Lactobacillus DSM15526 (Bifodan), Lactobacillus rhamnosus GG (LGG) (ATCC 53103) are described in U.S. Pat. No. 5,032,399, Lactobacillus rhamnosus LC705 (DSM 7061), propionic acid bacteria such as Propionibacterium freudenreichii subsp. shermanii PJS (DSM 7067), which are described in more detail in Finnish Patent Invention No. 92498 (Valio Oy), Nitrosomonas eutropha eutropha D23 (ABIome), Staphylococcus hominis A9 strain, C2 strain, AMT2 strain, AMT3 strain, AMT4-C2 strain, AMT4-Gl strain, and / or AMT4-D12 strain. (all manufactured by Matrisys Bioscience), Staphylococcus epidermidis strains M034, M038, All, AMT1, AMT5-C5, and / or AMT5-G6 (all manufactured by 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.

[0120] In one embodiment of the present invention, the composition comprises at least one bacterial strain as defined herein in combination with at least one strain selected from the group of lactic acid bacteria capable of improving the integrity of tight junctions.

[0121] In a further 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 jensenii P63 (DuPont), Bifidobacterium lactis 420 (DuPont) and Lactobacillus acidophilus La-14 (DuPont), and / or cell lysates and / or soluble metabolites of said probiotic strains.

[0122] Compositions according to the present invention suitable for oral consumption may contain 1 x 10 per serving or dose. 6 ~1×10 14 colony forming units (CFU) can be provided.

[0123] As used herein, and as well understood in the art, "treatment" is an approach for obtaining beneficial or desired results, including clinical results. For purposes of this context, beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms, reduction in the extent of disease, stabilization (i.e., non-worsening) of disease, prevention of disease, delay or slowing of disease progression, and / or remission or alleviation of the disease state.

[0124] The reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% reduction in the severity of the complication or symptom.

[0125] In one embodiment of the present invention, there may be provided a method of treating mammalian skin, comprising administering to a subject (e.g., a mammal) in need thereof a therapeutically effective amount of at least one bacterial strain, thereby treating the skin to reduce colonization and / or carrier levels and / or infection.

[0126] In a further embodiment of the invention, the skin colonization may be caused by an antibiotic-resistant microorganism. The skin colonization may be caused by MRSA.

[0127] The at least one bacterial strain may be capable of growing and colonizing the gastrointestinal tract, nasal cavity or skin of a mammal.

[0128] The present invention may offer several advantages. In particular, to the extent that antibiotic use has deleterious effects due to the potential for the production of antibiotic-resistant microbial species, it is desirable to provide antimicrobial treatment that does not utilize traditional antimicrobial agents. Therefore, the present invention does not contribute to the production of future generations of antibiotic-resistant pathogens.

[0129] 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 include a rinsing step to remove biofilm from the surface before adding the at least one lactic acid bacterium.

[0130] Deposit of biomaterials The following biological materials, microorganisms, have been deposited at the German Collection for Microorganisms and Cell Cultures: -Weissella viridescens LB10G, deposited as DSM32906; - Lactobacillus paracasei LB113R, deposited as DSM32907; - Lactobacillus plantarum LB244R, deposited as DSM32996; - Lactobacillus paracasei LB116R, deposited as DSM32908; Enterococcus faecium LB276R, deposited as DSM32997; - Lactobacillus plantarum LB316R, deposited as DSM33091; -Leuconostoc mesenteriodes LB349R, deposited as DSM33093; - Lactobacillus plantarum LB356R, deposited as DSM33094; - Lactobacillus plantarum LB312R, deposited as DSM33098; is.

[0131] It should be noted that embodiments and features described in the context of one of the aspects of the invention also apply to the other aspects of the invention.

[0132] The present invention will now be described in further detail in the following non-limiting examples. [Example]

[0133] Example 1 Strain Screening Identification

[0134] sample A lactic acid bacteria (LAB) strain collection was established for the identification and selection of bacterial strains according to the present invention. Samples from different sources, including homemade sauerkraut, kimchi, and healthy human donor samples (vaginal, oral, anal, and skin), were collected for the isolation of at least 995 lactic acid bacteria. The samples were collected on Man Rogosa Sharp (MRS, Sigma-Aldrich) broth and agar and anaerobically cultured at 37°C overnight or until colonies formed. Isolates were plated and subcultured to obtain pure colonies. Pure colonies were stored in MRS broth containing 25% glycerol at -80°C for future use. Strains were identified using standard 16S rRNS Sanger sequencing.

[0135] Example 2 Co-culture assay / competition assay

[0136] The competition between the bacterial strains and Staphylococcus aureus was evaluated based on 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).

[0137] The test organisms used were S. aureus subsp. aureus 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). S. aureus was cultured in Brain Heart Infusion (BHI) broth. S. aureus CC1 is a clonal type specifically associated with atopic dermatitis and associated with FLG mutations in patients with atopic dermatitis.

[0138] The cell density of overnight cultures of S. aureus and bacterial isolates was adjusted to an optical density at 600 nm (OD600) of 1 and harvested by centrifugation (6,000 rpm for 2 minutes). The cell pellets were washed twice in phosphate-buffered saline (1x PBS) and resuspended in 1x PBS. One milliliter of each cell suspension was mixed in 50 mL of BHI broth and co-incubated at 37°C for 24 hours. Monocultures of each S. aureus and each LAB were used as controls. Serial dilutions of the cell solution were plated onto nutrient agar plates at 0, 2, 6, 10, and 24 hours to count the colonies formed. MRS agar was used for LAB isolates, and mannitol salt phenol red agar (Sigma-Aldrich) was used for S. aureus.

[0139] Lactobacillus rhamnosus LGG (Chr. Hansen) was used as the control probiotic strain in all experiments.

[0140] Twenty-two strains of lactic acid bacteria (LAB) from a collection of 675 bacterial strains were identified as able to outgrow all three tested Staphylococcus aureus strains, as determined by their ability to reduce the growth of the test strains by at least 25%. Eight bacterial strains were determined to reduce growth by more than 90%. Lactobacillus rhamnosus LGG was unable to outgrow any S. aureus strains.

[0141] Example 3 Co-aggregation

[0142] Coaggregation was measured by a known method, Cisar, J.O. 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.

[0143] Inocula for all bacterial strains were grown in MRS broth, and S. aureus strains were grown in BHI broth overnight at 37°C. The overnight cell samples were harvested by centrifugation (6000 rpm for 2 minutes) and the supernatant was removed from the pellet. The pellet was washed twice in 1x PBS buffer.

[0144] The cell pellet was resuspended in 1x PBS, and 500 μl of Staphylococcus aureus and bacterial strains were dispensed into a 24-well plate. The plate was incubated on a shaker (200 rpm). The formation of autoagglutination and coagglutination was visually observed after 1 hour, 2 hours, 3 hours, and 24 hours.

[0145] As a control for autoagglutination, samples of S. aureus and bacterial strains were mixed with 750 ul of PBS buffer, resulting in a final volume of 1500 ul in each well.

[0146] The plates were incubated on a shaker at approximately 200 rpm for 24 hours. Coaggregate formation is observed after 1, 2, 3 and 24 hours.

[0147] Coaggregate formation was visually scored from 1 to 5 using the following scale: 1: No aggregation 2: Visual inspection of initial aggregation 3: Formation of aggregates smaller than 0.5 mm 4: Formation of aggregates greater than 0.5 mm and less than 1 mm 5: Formation of aggregates larger than 1 mm

[0148] [Table 1]

[0149] Example 4 Spot on lawn assay

[0150] 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, MPet al. (2016) Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Spot-on lawn tests for growth inhibition and antimicrobial metabolites were tested using methods described in Pathogenic Microorganisms, Frontiers in Microbiology 7(APR):1-10. https: / / doi.org / 10.3389 / fmicb.2016.00464.

[0151] Bacterial strain isolates from Example 1 were cultured from stock samples into 2 mL of MRS broth in 24-well plates. S. aureus test strains were cultured in approximately 200 mL of BHI broth in Erlenmeyer flasks. LAB isolates and S. aureus solutions were grown overnight at 37°C. The cell density of the overnight culture of S. aureus was increased to 600 nM in BHI broth. The cell suspension was adjusted to an optical density (OD600) of 1 at 100 μL and then diluted to a 10-2 dilution in PBS buffer. 200 microliters of the cell suspension was spread on a BHI agar plate. The S. aureus lawn-containing plate was allowed to dry in sterile air for approximately 10-20 minutes. Triplicate 20 μL of isolated LAB was spotted onto the S. aureus lawn. The plate was left to dry and then aerobically incubated at 37°C overnight. The plate was photographed, and the zone of inhibition was measured in mm as the clearing area around the spot. Growth inhibition was observed as the ability of the bacterial strain to overgrow the Staphylococcus strain within the spot area, indicated in Table 2 as (+). If the Staphylococcus strain was able to outgrow the spotted bacterial strain, no growth inhibition was detected, indicated in Table 2 as (-).

[0152] Five LAB strains were identified as having significant growth inhibitory effects against Staphylococcus aureus and also zones of inhibition greater than 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 staphylococcal test strains, nor was Lactobacillus rhamnosus LGG able to confer any zones of clearance.

[0153] [Table 2]

[0154] Significant inhibition of CC1 was determined, and this clonal complex of S. aureus 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).

[0155] Lactobacillus acidophilus NCFM, Lactobacillus salivarius Ls-33, Bifidobacterium lactis 420, Lactobacillus acidophilus La-14, and Propionibacterium yensenii P63 (all commercially available from DuPont) have all been implicated as potentially ameliorating disorders associated with tight junction function, such as atopic dermatitis, by improving skin barrier function. However, in spot-on lawn tests with CC1, none of these commercially available probiotic strains was able to inhibit CC1 growth and therefore prevent or treat staphylococcal infections.

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Claims

1. The group consisting of: - Weissella viridescens LB10G, deposited under DSM 32906; - Lactobacillus paracasei LB113R, deposited under DSM 32907; - Lactobacillus plantarum LB244R, deposited under DSM 32996; - Lactobacillus paracasei LB116R, deposited under DSM 32908; - Enterococcus faecium LB276R, deposited under DSM 32997; - Lactobacillus plantarum LB316R, deposited under DSM 33091; - Leuconostoc mesenteriodes LB341R; - Leuconostoc mesenteriodes LB349R, deposited under DSM 33093; - Lactobacillus plantarum LB356R, deposited under DSM 33094; - Lactobacillus plantarum LB312R, deposited under DSM 33098; A bacterial strain having at least 95% genetic homology to one or more of the bacterial strains selected from:

2. The genetic homology is - Weissella viridescens LB10G, deposited under DSM 32906; - Lactobacillus paracasei LB113R, deposited under DSM 32907; - Lactobacillus plantarum LB244R, deposited under DSM 32996; - Lactobacillus paracasei LB116R, deposited under DSM 32908; - Enterococcus faecium LB276R, deposited under DSM 32997; - Lactobacillus plantarum LB316R, deposited under DSM 33091; - Leuconostoc mesenteriodes LB341R; - Leuconostoc mesenteriodes LB349R, deposited under DSM 33093; - Lactobacillus plantarum LB356R, deposited under DSM 33094; - Lactobacillus plantarum LB312R, deposited under DSM 33098; 2. The bacterial strain of claim 1, which is at least 96%, such as at least 97%, for example 98%, such as at least 99%, at least 99.5%, such as at least 99.8%, for example at least 99.9%, such as 100% (identity), to one of the bacterial strains selected from the group consisting of:

3. The bacterial strain - Weissella viridescens LB10G, deposited under DSM 32906; - Lactobacillus paracasei LB113R, deposited under DSM 32907; - Lactobacillus plantarum LB244R, deposited under DSM 32996; - Lactobacillus paracasei LB116R, deposited under DSM 32908; - Enterococcus faecium LB276R, deposited under DSM 32997; - Lactobacillus plantarum LB316R, deposited under DSM 33091; - Leuconostoc mesenteriodes LB341R; - Leuconostoc mesenteriodes LB349R, deposited under DSM 33093; - Lactobacillus plantarum LB356R, deposited under DSM 33094; - Lactobacillus plantarum LB312R, deposited under DSM 33098; 3. The bacterial strain according to claim 1, selected from the group consisting of:

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

5. The bacterial strain according to any one of claims 1 to 4, wherein the bacterial strain is an isolated bacterial strain.

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

7. 6. The composition of claim 5, wherein one or more of the bacterial strains is provided as one or more viable strains, one or more killed or inactivated strains, one or more strain lysates, one or more strain metabolites, or a combination thereof.

8. 8. The composition of claim 6, wherein the composition is an emulsion, oil, gum, 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 concentration of said bacterial strains, preferably one or more viable strains and / or one or more dead strains, is 10 ５ ~10 １３ Colony forming units (CFU) range, e.g., 10 ９ ~10 １１ CFU range, etc., 10 ７ ~10 １２ 10, such as the range of CFU ３ ~10 １４ The composition of any one of claims 6 to 9, wherein the range is CFU.

11. 9. The composition according to any one of claims 5 to 8, wherein the concentration of said bacterial strains, preferably one or more killed / inactivated strains, one or more strain lysates, one or more strain metabolites is at a concentration of 0.001% (w / w) to 20% (w / w).

12. The composition of any one of claims 6 to 11, wherein the composition further comprises a prebiotic composition.

13. The composition of any one of claims 6 to 12, wherein the bacterial strain of claims 1 to 5 is the only bacterium present in the composition.

14. 13. The composition of any one of claims 5 to 12 for use in treating, alleviating, inhibiting, preventing and / or preventing the growth of pathogenic microorganisms.

15. 15. The composition of any one of claims 6 to 14 for use in treating, alleviating, suppressing and / or preventing one or more pathogenic bacterial infections in a mammal.

16. 16. The composition of any one of claims 6 to 15 for use in treating, alleviating, suppressing and / or preventing one or more antibiotic-resistant bacterial infections in a mammal.

17. The composition of any one of claims 14 to 16, wherein the bacterial infection is a staphylococcal infection in a mammal.

18. 18. The composition of claim 17, wherein the staphylococcal infection is an MRSA infection.

19. 19. The composition of any one of claims 14 to 18 for use in the treatment, alleviation, suppression and / or prevention of diseases resulting from staphylococcal infections, such as dermatitis, atopic dermatitis, eczema, carbuncles, cellulitis, rosacea, psoriasis, diaper rash, impetigo or wounds.

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