Strains, compositions and methods of use

Abstract

The present invention relates to a composition comprising one or more probiotic bacterial strains, which composition can partially or completely inhibit the growth of one or more fungal species.

Description

Technical Field

[0001] The present invention relates to novel lactic acid bacterial strains that can be used alone or in combination as probiotics. In particular, the present invention relates to new strains, new fungicidal compositions and the use of the strains for the prevention or treatment of fungal diseases.

Background Art

[0002] The incidence of fungal infections has increased significantly in the past few decades, mainly due to the increasing antimicrobial resistance and the limited number of effective antifungal drugs, which still have many side effects. The use of broad-spectrum antibiotics, denture stomatitis, catheters and parenteral nutrition, the presence of immunosuppression, the disruption of the mucosal barrier, as well as the use of chemotherapy and radiotherapy and the dysfunctional microbiome are among the most important predisposing factors for the development of invasive fungal infections.

[0003] The pathogenicity of fungal species is due to certain pathogenic factors such as the ability to evade host defenses, hyphal formation, adhesion and biofilm formation (on host tissues or medical devices), and the production of tissue-damaging hydrolases such as proteases, phospholipases and hemolysins.

[0004] A biofilm is a highly organized biological community in which microorganisms form a structured, coordinated functional community embedded in an extracellular matrix they produce themselves. Biofilm production is also associated with a high level of antimicrobial resistance in the related organisms. The ability of fungal species to form drug-resistant biofilms is an important factor in the contribution of fungal species to human diseases, but it is also recognized that the more general availability of new medical practices (diseases caused by immunosuppressive therapy, invasive surgical procedures, the use of broad-spectrum antibiotics and the dysfunctional microbiome) is also very important. Biofilms are resistant to various antifungal agents currently in clinical use, including amphotericin B and fluconazole, and there appear to be multiple resistance mechanisms.

[0005] Antifungal drugs exhibit activity by either killing fungal cells, for example by affecting substances in the cell wall, causing leakage and death of the cell contents, or preventing the fungal cells from growing and regenerating. There are many antifungal classes: polyenes including amphotericin, nystatin and pimaricin; azoles including fluconazole, itraconazole, ketoconazole, miconazole, voriconazole, posaconazole and ravuconazole; echinocandins such as caspofungin and micafungin; and allylamines including naftifine, terbinafine, morpholine drugs, amorolfine and griseofulvin; as well as antimetabolite antifungal drugs incorporating 5-fluorocytosine. What these drugs have in common is that problems associated with the development of resistance, lack of efficiency and toxicity of the compounds are increasing.

[0006] Amphotericin B and fluconazole are among the most widely used antifungal agents for treating systemic fungal infections. The former is restricted in its use because of its high degree of toxicity in humans. The latter is frequently prescribed for treating infections, but a large number of fungal species exhibit fluconazole resistance.

[0007] Fungal diseases are infections caused by any fungus that invades tissues in humans and animals, causing superficial, subcutaneous, or systemic diseases. Many different types of fungi can cause fungal diseases, and some types, such as Cryptococcus and Histoplasma, can cause severe, life-threatening infections. Fungal diseases are infectious diseases caused by microscopic fungi. Fungi reproduce in skin folds, fingernails, hair, and mucous membranes such as the mouth, depending on the type. There are two main types of fungi that cause lesions and can be confused with, for example, eczema. Dermatophytes: These fungi attack the keratin found in the skin, fingernails, and hair. These actual parasites are transmitted by contact with animals, humans, or surfaces. Yeasts (Candida): They occur in mucous membranes and can initiate overgrowth. As a result, lesions occur on the skin, usually around the openings. The most common example occurs in the diaper area of infants. Fungal skin infections can occur anywhere on the body. Some of the most common ones are athlete's foot, jock itch, ringworm, and yeast infections.

[0008] Superficial fungal infections, also known as dermatophytosis, are limited to the skin and are caused by Microsporum, Trichophyton, or Epidermophyton. For example, athlete's foot is caused by Trichophyton or Epidermophyton. Subcutaneous infections spread to the tissues under the skin, including adjacent structures such as bone and organs, are rare, and are often chronic. Candidiasis (caused by Candida) can be a superficial infection (such as thrush or vaginitis) or a disseminated infection that affects certain target organs such as the eye or kidney. In sporotrichosis (caused by Sporothrix), painful ulcers and nodules appear in the subcutaneous tissue. In systemic fungal infections, the fungus can invade normal or immunosuppressed hosts (causing opportunistic infections). Cryptococcosis (caused by Cryptococcus) and histoplasmosis (caused by Histoplasma) are characterized by respiratory distress.

[0009] Athlete's foot, also known as tinea pedis, is a fungal infection of the feet. Fungi grow best in warm, moist places such as shoes, socks, swimming pools, locker rooms, and public showers. Fungi are often seen in summer and in hot, humid climates. Athlete's foot occurs more frequently in people who wear tight shoes, do not change wet or sweaty socks, and use public baths and pools.

[0010] A type of fungal infection called tinea cruris causes jock itch. This infection is also known as groin ringworm. Tinea cruris prefers warm, moist areas such as the genitals, inner thighs, and buttocks, and fungal infections occur more frequently in summer or in warm, humid climates.

[0011] Ringworm is often a red, itchy rash that is ring-shaped. Ringworm is very mildly contagious. Ringworm can spread from person to person through direct contact or indirectly via objects that have the fungus on them.

[0012] Tinea corporis, also known as body ringworm, is a fungal skin infection, not an infection by worms. The name "ringworm" comes from the ring-shaped rash with a wavy, worm-like border. Tinea corporis can spread through direct contact with infected people or animals. It can also be transmitted from clothing or furniture. Heat and humidity can contribute to spreading the infection.

[0013] Mycoses are classified as superficial, cutaneous, subcutaneous, or systemic (deep) infections, depending on the type and extent of tissue damage and the host response to the pathogen.

[0014] Cutaneous mycoses (superficial mycoses) are fungal diseases that are limited to the outer layer (stratum corneum) of the skin, nails, and hair, and rarely invade deeper tissues or internal organs. The infection is limited to the stratum corneum, and there is little or no tissue reaction. Superficial mycoses include the following fungal infections and their causative agents: black piedra (Piedraia hortae), white piedra (Trichosporon beigelii), pityriasis versicolor (Malassezia furfur), and tinea nigra (Phaeoannellomyces werneckii). Pityriasis versicolor is a common superficial mycosis characterized by hypopigmentation or hyperpigmentation of the skin of the neck, shoulders, chest, and back. Pityriasis versicolor is caused by Malassezia furfur, which involves only the superficial keratin layer. Black piedra is a superficial mycosis caused by Piedraia hortae, presenting small hard black nodules involving the hair shaft. In comparison, white piedra caused by T. beigelii is characterized by soft, fragile beige-colored nodules at the distal ends of the hair shafts. Tinea nigra most typically appears as a brown to black silver nitrate-like stain on the palms or soles of the feet.

[0015] Cutaneous Mycoses can be classified as dermatophytoses or cutaneous mycoses (dermatomycoses) and general infections. Dermatophytoses are caused by genera such as Arthroderma, Lophophyton, Nannizzia, Epidermophyton, Microsporum, Deb arIt is caused by pathogens of the genera Epidermophyton, Kluyveromyces, and Trichophyton. Dermatomycoses are skin infections caused by other fungi, the most common of which are species of Candida and Pichia. Dermatophytoses are characterized by genus-specific anatomical site specificities. For example, Epidermophyton floccosum infects only the skin and nails and does not infect the hair shaft and hair follicles. In contrast, species of Microsporum infect the hair and skin but do not involve the nails. Species of Trichophyton can infect the hair, skin, and nails.

[0016] There are three general types of subcutaneous mycoses: chromoblastomycosis, mycetoma, and sporotrichosis. All appear to be caused by traumatic inoculation of pathogenic fungi into the subcutaneous tissue. Chromoblastomycosis is a subcutaneous mycosis characterized by verrucoid lesions of the skin (usually on the lower extremities).

[0017] Chromoblastomycosis and mycetoma are caused by only certain fungi. The most common causes of chromoblastomycosis are Fonsecaea pedrosoi, Fonsecaea compacta, Cladosporium carionii, and Phialophora verrucosa.

[0018] The causes of mycetomas are more diverse and can be classified into fungal and actinomycotic mycetomas. A common fungus in fungal mycetomas is Pseudallescheria boydii, and the most common cause of actinomycotic mycetomas is Nocardia brasiliensis. These fungi can cause a variety of infections, from superficial to subcutaneous and deep (visceral) infections, characterized by the presence of black hyphae and / or yeast-like cells in the tissue. Such deep infections caused by black fungi are called phaeohyphomycosis.

[0019] Sporotrichosis is the third common class of subcutaneous mycosis. This infection is caused by Sporothrix schenckii and involves the subcutaneous tissue at the site of traumatic inoculation. This infection usually spreads along the cutaneous lymphatic channels of the involved extremity.

[0020] Deep mycosis is caused by primary pathogenic and opportunistic fungal pathogens. Primary pathogenic fungi can establish infection in normal hosts, while opportunistic pathogens require a susceptible host to establish infection (e.g., cancer, organ transplantation, surgery, and AIDS). Primary deep pathogens usually gain access to the host via the respiratory tract. Opportunistic fungi that cause deep mycosis invade via the respiratory tract, gastrointestinal tract, or intravascular devices.

[0021] Primary systemic fungal pathogens include Coccidioides immitis, Histoplasma capsulatum, Blastomyces dermatitidis, and Paracoccidioides brasiliensis. Opportunistic fungal pathogens include Cryptococcus neoformans, Candida spp., Aspergillus spp., Penicillium marneffei, Zygomycetes, Trichosporon beigelii, and Fusarium spp.

[0022] Topical antifungal agents, commonly available as creams, liquids, or sprays, are often ineffective against superficial infections. Antifungal griseofulvin has had some success in the treatment of superficial mycosis, and amphotericin B and flucytosine are used in the treatment of subcutaneous and systemic mycosis. However, an increasing number of mycosis are difficult to treat.

[0023] Athlete's foot is an infection caused by a type of fungus known as dermatophytes. It can only infect the outermost layer of dead keratin, and dermatophytes affect the skin, hair shafts, and nails. Dermatophytes are classified into three genera: Trichophyton, Microsporum, and Epidermophyton. T. rubrum is the most commonly associated dermatophyte with athlete's foot. Other dermatophytes can also cause this condition, but are less frequently isolated from humans. Fungal spores from T. rubrum can survive for 12 months in human scales and are thus easily transmitted from person to person in locker rooms and public showers.

[0024] Antifungal resistance is complex and multifaceted. Antifungal resistance can be inducible in response to a compound or can be an irreversible genetic change resulting from long-term exposure. Specifically, these include changes or overexpression of target molecules, active efflux through efflux pumps, limited diffusion, tolerance, and cell density, all of which are characterized mechanisms utilized by fungi to counteract the effects of antifungal treatment. Planktonic cells generally rely on irreversible genetic changes to maintain the resistant phenotype, whereas biofilm cells can survive due to their physical presence and population density, conferring a nearly inducible resistant phenotype independent of defined genetic changes.

[0025] Fungal species are part of the normal microbiota of the oral cavity, skin, gastrointestinal tract, and vaginal tract and are responsible for several clinical manifestations, from mucocutaneous overgrowth to bloodstream infections. Over the years, the pathological conditions caused by the fungal microbiota are more likely to recur and are difficult to treat, especially when the patient shows some level of immunosuppression, is receiving antibiotics, or has a dysfunctional microbiome with reduced microbial diversity, for example.

[0026] Lactic acid bacteria are part of the human gastrointestinal tract, oral, and vaginal microbial flora. Lactic acid bacteria play an important role in protecting the human body from infection through the production of acids and other antibacterial products such as hydrogen peroxide H 2 O 2 , antibacterial peptides, or biosurfactants.

[0027] Many topical, vaginal, oral, and systemic drugs can kill lactic acid bacteria. Thus, treatment of infections with antibiotics can increase the risk of recurrent acquisition of the infection in the body or of new infections caused by resistant microorganisms, such as fungi.

[0028] For decades, antifungal agents have been successfully used to prevent mucosal and invasive fungal infections. However, due to drug side effects (nausea, vomiting, and diarrhea) and the emergence of resistant strains, antifungal prophylaxis has not been completely successful. The limited efficacy and the gradual emergence of resistance to antifungal drugs are concerns, and thus alternative treatments are urgently needed.

[0029] The use of probiotic microorganisms against fungal diseases could be an attractive alternative treatment for treating or preventing fungal diseases considering the limitations of currently available antibacterial compounds.

[0030] Lactic acid bacteria for vaginal or oral use have been available for over 50 years in the form of probiotic preparations available in health food stores or as dairy products. Typically, traditional probiotics are used to maintain a healthy gut flora and are not known to have any targeted or specific antibacterial mechanisms.

[0031] The present invention provides novel probiotic strains and compositions that target at least one pathogenic fungus, preferably a fungus causing mycosis, and in particular the compositions inhibit the growth of dermatophytes and exhibit general antifungal activity that causes growth inhibition, inhibition of spore germination, inhibition of hyphal formation, inhibition of biofilm attachment or formation, or the ability to prevent the potential of the fungus to secrete pathogenic factors as phospholipases, proteinases, and hemolytic factors, and / or can prevent and / or treat mycosis in humans or animals. SUMMARY OF THE INVENTION

[0032] Accordingly, the object of the present invention relates to novel lactic acid bacteria, compositions, topical compositions, vaginal compositions, ocular compositions, otic compositions, rectal compositions or oral compositions containing the novel lactic acid bacteria, and their ability to inhibit the growth of dermatophytes, inhibit growth, inhibit spore germination, inhibit hyphal formation, inhibit the growth of fungi, and / or prevent or treat mycosis.

[0033] In particular, it is an object of the present invention to provide novel lactic acid bacterial strains and compositions containing such novel lactic acid bacterial strains that solve the above-mentioned problems of the prior art, particularly having antifungal activity and thereby having the ability to prevent mycosis, preferably without the above-mentioned side effects.

[0034] Accordingly, one aspect of the present invention relates to a composition comprising Lactiplantibacillus plantarum, LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum, LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum, LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S; or a combination thereof.

[0035] Yet another aspect of the present invention relates to a composition comprising one or more probiotic bacterial strains, which composition is capable of partially or completely inhibiting the growth of one or more fungal species, in particular one or more dermatophytes.

[0036] A further aspect of the present invention is - Lactiplantibacillus plantarum, LB990R, deposited under accession number DSM 34494; and / or - Lactiplantibacillus plantarum, LB681R, deposited under accession number DSM 34250; and / or - Lacticaseibacillus paracasei LB857R, Lactobio deposited under accession number DSM 34493; and / or - Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; and / or - Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249; relating to an isolated probiotic bacterial strain selected from and / or A further aspect of the present invention is capable of partially or completely inhibiting the growth of one or more dermatophytes, - Lactiplantibacillus plantarum LB990R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34494; - Lactiplantibacillus plantarum LB681R, deposited on May 5, 2022 with accession number DSM 34250 by Lactobio A / S; - Lacticaseibacillus paracasei LB857R, deposited on January 10, 2023 with accession number DSM 34493 by Lactobio A / S; - Lactiplantibacillus plantarum LB760R, deposited on January 10, 2023 with accession number DSM 34492 by Lactobio A / S; - Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited on May 5, 2022 with accession number DSM 34249 by Lactobio A / S. It relates to an isolated probiotic bacterial strain selected from

[0037] Another aspect of the present invention is - The treatment or prevention of mycosis, preferably the treatment and / or prevention of mycosis in humans or animals; - The treatment of dermatophytosis, preferably the treatment and / or prevention of dermatophytosis in humans or animals; and / or The prevention, inhibition and / or treatment of fungal growth in crops, seeds, food or feed for use in Compositions comprising Lactiplantibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus paracasei LB857R, deposited under accession number DSM 34493; or combinations thereof are provided.

[0038] Yet another aspect of the invention relates to compositions comprising one or more bacterial strains selected from one or more lactic acid bacteria for use in the prevention and / or treatment of mycobiota dysbiosis caused by fungi.

[0039] Yet another aspect of the invention relates to compositions comprising one or more bacterial strains selected from one or more lactic acid bacteria for use in the prevention and / or treatment of mycosis in humans or in animals.

[0040] Another aspect of the present invention relates to a composition comprising one or more bacterial strains selected from Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249 or combinations thereof for use in the prevention and / or treatment of fungal diseases in humans or animals.

[0041] Another aspect of the present invention relates to a composition comprising one or more bacterial strains selected from one or more lactic acid bacteria for use in the prevention and / or treatment of fungal infections or contaminations in crops, food or animal feed.

[0042] A further aspect of the present invention relates to an antifungal composition comprising one or more probiotic bacterial strains for inhibiting fungal spore germination and / or inhibiting fungal hyphal formation.

[0043] Another aspect of the present invention relates to the use of a composition according to the present invention comprising one or more bacterial strains selected from Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249 or combinations thereof for the prevention, inhibition or treatment of fungal growth in crops, seeds, food or feed.

[0044] A further aspect of the present invention relates to a composition comprising one or more bacterial strains selected from one or more lactic acid bacteria for use in the prevention and / or treatment of mycosis in humans or animals.

[0045] A further aspect of the present invention is a method for reducing and / or inhibiting fungal spore germination and / or fungal hyphal formation, comprising adding a composition according to the present invention to a surface infected with a fungus or preventing the surface from being infected with a fungus.

[0046] Yet another aspect of the present invention is a method for preventing mycosis in the environment, comprising the step of administering to the environment an effective amount of a lactic acid bacterium having antifungal activity, wherein the environment is a dwelling, workplace, laboratory, industrial environment, hospital environment, aquatic environment, medical device, dental device, plant, corps, epithelial cell, mucosa, animal or human body.

[0047] Yet another aspect of the present invention is a method for preventing the growth of fungi in an environment, the method comprising the step of administering to the environment an effective amount of a lactic acid bacterium having antifungal activity, wherein the environment is selected from a dwelling, a workplace, a laboratory, an industrial environment, an aquatic environment, a medical device or a dental device.

[0048] A further aspect of the present invention relates to a food or feed ingredient or preservative, and a personal hygiene product, comprising the composition according to the present invention. Detailed Description of the Invention

[0049] Accordingly, the inventors of the present invention have found certain lactic acid bacteria having antifungal activity that target pathogenic fungi.

[0050] The lactic acid bacteria of the present invention are thought to be able to inhibit or prevent mycosis.

[0051] A preferred embodiment of the present invention is a composition comprising one or more probiotic bacterial strains, the composition being capable of inhibiting, partially or completely, the growth of one or more fungal species.

[0052] A further preferred embodiment of the present invention is a composition comprising one or more probiotic bacterial strains, the composition being capable of inhibiting, partially or completely, the growth of one or more dermatophytes.

[0053] The one or more dermatophytes are preferably selected from Arthroderma, Lophophyton, Nannizzia, Epidermophyton, Microsporum, Deb ar yomyces, Kluyveromyces, Pichia, Trichophyton, or combinations thereof.

[0054] Preferably, the probiotic bacterial strain can be selected from at least one lactic acid bacterial strain.

[0055] One or more probiotic bacterial strains can preferably be one or more isolated probiotic bacterial strains.

[0056] At least one lactic acid bacterial strain can preferably be selected from one or more Lacticaseibacillus bacterial strains; and / or one or more strains selected from one or more Lactiplantibacillus bacterial strains; or a combination thereof.

[0057] Preferably, one or more probiotic bacterial strains can be selected from Lacticaseibacillus paracasei and / or Lactiplantibacillus plantarum.

[0058] One or more lactic acid bacteria strains are Lactiplantibacillus plantarum LB990R, deposited by Lactobio A / S on January 10, 2023, with accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited by Lactobio A / S on May 5, 2022, with accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited by Lactobio A / S on January 10, 2023, with accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited by Lactobio A / S on January 10, 2023, with accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited by Lactobio A / S on May 5, 2022, with accession number DSM 34249; or can be selected from combinations thereof.

[0059] In one embodiment of the present invention, the composition can be a composition comprising one or more lactic acid bacteria according to the present invention, together with an acceptable carrier and / or diluent.

[0060] Preferably, the composition contains lactic acid bacteria according to the present invention at 10 3 ~10 13 colony forming units per gram. More specifically, the composition contains lactic acid bacteria at 10 4 ~10 12 colony forming units per gram. More specifically, the composition contains lactic acid bacteria at 10 5 ~10 11 colony forming units per gram.

[0061] The composition can be in the form of a suspension, spray, gel, cream, lotion, powder, capsule, oil, solution for lavages, ovules, vaginal insert, suppository, troche, tablet, microencapsulated product, or in the form of a dietary supplement or food.

[0062] In one embodiment of the present invention, the composition can be a pharmaceutical composition comprising one or more lactic acid bacteria according to the present invention, together with a pharmaceutically acceptable carrier and / or diluent.

[0063] Preferably, it is a pharmaceutical composition containing lactic acid bacteria according to the present invention at 10 3 ~10 13 colony forming units per gram. More specifically, it is a pharmaceutical composition containing lactic acid bacteria at 10 6 ~10 12 colony forming units per gram. More specifically, it is a pharmaceutical composition containing lactic acid bacteria at 10 7 ~10 11 colony forming units per gram.

[0064] The pharmaceutical composition can be in the form of a suspension, spray, gel, cream, lotion, powder, capsule, oil, solution for lavages, ovules, vaginal insert, suppository, troche, tablet, bandage, plaster, microencapsulated product, or in the form of a dietary supplement or food.

[0065] In one embodiment of the present invention, the composition can be a dermatological composition comprising one or more lactic acid bacteria according to the present invention, together with a dermatologically acceptable carrier and / or diluent.

[0066] Preferably, it is a dermatological composition containing lactic acid bacteria according to the present invention at 10 3 ~10 13 colony forming units per gram. More specifically, it is a dermatological composition containing lactic acid bacteria at 10 6 ~10 12 colony forming units per gram. More specifically, it is a dermatological composition containing lactic acid bacteria at 10 7 ~10 11A dermatological composition containing lactic acid bacteria in colony forming units.

[0067] The dermatological composition can be in the form of a suspension, spray, gel, cream, powder or lotion.

[0068] The composition contains a therapeutically effective amount of lactic acid bacteria, in particular a therapeutically effective amount of - Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S; or - Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S; or - Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S; or - Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S; or - Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S; or - Combinations thereof One or more bacteria selected from the above can be provided.

[0069] The composition may be provided with a therapeutically effective amount of Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S, in further combination with at least one additional lactic acid bacterium, or a combination thereof.

[0070] In the context of the present invention, the terms "therapeutically effective amount" or "effective amount" may refer to the amount of a compound in a composition or preparation that, when administered as part of a desired dosing regimen (to a human or an animal, preferably a human), alleviates symptoms, improves a condition, or delays the onset of a disease state, according to clinically acceptable criteria for the disorder or condition to be treated or for cosmetic purposes, e.g., at a reasonable benefit-risk ratio applicable to any medical treatment.

[0071] Another embodiment of the present invention is - the treatment or prevention of fungal infections, preferably the treatment and / or prevention of fungal infections in humans or animals; -Treatment of dermatophytosis, preferably treatment and / or prevention of dermatophytosis in humans or animals; and / or Prevention, inhibition and / or treatment of fungal growth in crops, seeds, food or feed For use in Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; or a composition comprising a combination thereof.

[0072] For actual use, the microorganisms of the present invention can be formulated in suitable dosage forms such as suspensions, sprays, gels, creams, lotions, powders, capsules, oils, solutions for lavages, ovules, vaginal inserts, suppositories, troches, tablets, microencapsulated products, or nutritional supplements or foods. The unit dose can contain each single strain of 10-10 13 cells, and the preferred dosage is more than 10 4 cells per unit dose. A more preferred dosage is more than 10 5 cells per unit dose.

[0073] The bacterial cultures can be stabilized in freeze-dried, lyophilized or microencapsulated form and can be prepared according to conventional methods.

[0074] For the preparation of aqueous formulations for washing and irrigation, a two-phase system can be used. For example, a small bottle equipped with a reservoir containing freeze-dried microorganisms dissolved in a suitable liquid carrier contained in the bottle before use.

[0075] For the preparation of creams and gels, lactic acid bacteria can be stabilized in the formulation. A formulation having a low water activity or in microencapsulated form is preferred.

[0076] For the preparation of capsule or tablet formulations for oral ingestion, lactic acid bacteria can be stabilized using capsule and tablet technologies known in the art for lactic acid bacteria stabilization, for example, the technologies used for probiotics as dietary supplements.

[0077] In one embodiment of the present invention, one or more lactic acid bacterial strains according to the present invention can be provided in a composition suitable for preventing and / or treating vaginal mycosis.

[0078] In a further embodiment of the present invention, one or more lactic acid bacterial strains according to the present invention can be provided in a composition for preventing and / or treating urinary tract infections.

[0079] In a further embodiment of the present invention, one or more lactic acid bacterial strains according to the present invention can be provided in a composition for preventing and / or treating local, skin, nail, eye, ear, oral, intestinal, gastrointestinal or mucosal infections.

[0080] In still a further embodiment of the present invention, one or more lactic acid bacterial strains according to the present invention can be provided in a composition for preventing and / or treating oral mycosis.

[0081] A further embodiment of the present invention is that the composition according to the present invention and / or one or more lactic acid bacteria can be used in combination with a medical device.

[0082] A further embodiment of the present invention relates to a method for treating or preventing microbial imbalance in the mucosa of humans or animals, comprising administering an effective amount of lactic acid bacteria according to the present invention.

[0083] In the context of the present invention, the term "prevent" may be recognized in the art and, when used in relation to conditions such as local recurrence, may be well understood in the art and includes administration of a composition that reduces the frequency of symptoms of a medical condition in a subject or delays its onset, compared to a subject not given the composition. Thus, prevention of an infection includes, for example, reducing the number of detectable pathogenic microorganisms in a population of patients / animals receiving prophylactic treatment compared to an untreated control population and / or delaying the appearance of detectable lesions in the treated population, for example, in a statistically and / or clinically significant amount, compared to an untreated control population.

[0084] As used herein, the term "treat" or "treatment" includes reversing, reducing, or halting the symptoms, clinical signs, and underlying pathology of a condition so as to improve or stabilize the condition of a subject.

[0085] As used herein and as well understood in the art, "treatment" can be an approach to obtain a beneficial or desired result, including clinical results. In this subject, beneficial or desired clinical results can include, but are not limited to, alleviation or improvement of one or more symptoms, reduction in the extent of a disease, a stabilized (i.e., non-worsening) state of a disease, prevention of a disease, delay in the progression or reduction in the rate of a disease, and / or improvement or remission of a disease state.

[0086] A reduction can be, for example, at least a 10% reduction, at least a 20% reduction, such as at least a 30% reduction, at least a 40% reduction, such as at least a 50% reduction, at least a 60% reduction, such as at least a 70% reduction, at least an 80% reduction, such as at least a 90% reduction, at least a 95% reduction, such as at least a 98% reduction, at least a 99% reduction, such as a 100% reduction in the severity of the complication or symptom, compared to the state before treatment.

[0087] Mycosis can be a fungal infection of humans and animals and can be caused by the invasion of human or animal tissue by one or more fungal species. Mycosis can be superficial mycosis, subcutaneous mycosis or systemic mycosis.

[0088] (The fungal species that cause mycosis) can be any fungal species that cause mycosis. In particular, the fungal species are Cryptococcus and Histoplasma, (which can cause severe, life-threatening infections), Candida, Microsporum, Trichophyton, or Epidermophyton, Sporothrix (mainly superficial mycosis), Piedraia hortae, Trichosporon beigelii, pityriasis versicolor, Phaeoannellomyces werneckii, Arthroderma, Lophophyton, Nannizzia, Epidermophyton, Microsporum, Deb arIt may be selected from the group consisting of Saccharomyces, Kluyveromyces, Fonsecaea pedrosoi, Fonsecaea compacta, Cladosporium carionii, Phialophora verrucose, Pseudallescheria boydii, Nocardia brasiliensis, Sporothrix schenckii, Coccidioides immitis, Histoplasma capsulatum, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Cryptococcus neoformans, Candida species, Aspergillus species, Penicillium marneffei, Zygomycetes, Trichosporon beigelii, and Fusarium species, or combinations thereof.

[0089] The composition according to the present invention can inhibit, prevent, or treat mycosis at any location in the human or animal body, such as on the skin, in skin folds, on nails, on fingernails, in the eyes, in the ears, in hair, on feet, on hands, and in mucous membranes such as the intestine, digestive tract, or mouth.

[0090] The composition according to the present invention may be suitable for inhibiting, preventing, or treating tinea infections such as tinea capitis, tinea corporis, tinea cruris, tinea faciei, tinea incognito, tinea manuum, tinea pedis, and onychomycosis.

[0091] The composition according to the present invention is - Tinea pedis, also known as athlete's foot, a fungal infection of the feet - Tinea cruris, also known as jock itch, a fungal infection of the groin - Tinea corporis, also known as ringworm, a fungal infection of the skin, which can appear as red, itchy, scaly, circular rashes and may cause hair loss in the affected area and may be suitable for inhibiting, preventing or treating tinea infections such as those described above.

[0092] In one embodiment of the present invention, the fungal species may not include Malassezia spp.

[0093] In a further embodiment of the present invention, the fungal species may not include Candida spp.

[0094] "Dermatophytosis" (superficial mycosis) is a fungal disease that can be localized to the outer layer (keratinized layer) of the skin, nails and hair.

[0095] In one embodiment of the present invention, the one or more probiotic bacterial strains can be one or more lactic acid bacterial strains.

[0096] The term "lactic acid bacteria" includes species of the families Lactobacillaceae, Aerococcaceae, Bifidobacteriaceae, Carnobacteriaceae, Enterococcaceae, Leuconostocaceae and Streptococcaceae. These are considered non-pathogenic and are generally used as probiotic bacteria to improve the gastrointestinal microbiota and in the treatment of gastrointestinal symptoms. The lactic acid bacteria according to the present invention can preferably be selected from lactic acid bacterial strains that inhibit, prevent and reduce the growth of mycotic fungi. In particular, mycosis in mammals.

[0097] The inventors of the present invention have surprisingly found certain strains of lactic acid bacteria that have been shown to be able to inhibit, prevent, or reduce the growth of fungi.

[0098] Preferably, the present invention relates to an antifungal composition comprising one or more probiotic bacterial strains for inhibiting the spore germination of fungi and / or inhibiting the hyphal formation of fungi.

[0099] Preferably, the fungus can be a dermatophyte.

[0100] In one embodiment of the present invention, the one or more probiotic bacterial strains are selected from one or more Lactiplantibacillus plantarum, one or more Lacticaseibacillus paracasei, or combinations thereof.

[0101] Preferably, the one or more probiotic bacterial strains can be one or more isolated probiotic bacterial strains.

[0102] In one embodiment of the present invention, the one or more probiotic bacterial strains are - Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; and / or - Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; and / or - Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; and / or - Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; and / or - Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249 - Combinations thereof. It may be selected from one or more of the above.

[0103] Preferred embodiments of the present invention are Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S; Or it may relate to a composition comprising a combination thereof.

[0104] The composition according to the present invention may contain live or dead cells of the lactic acid bacteria according to the present invention.

[0105] The composition according to the present invention may contain metabolites produced by the lactic acid bacteria of the present invention and / or actives obtained from the lactic acid bacteria of the present invention.

[0106] In one embodiment of the present invention, the composition contains a concentration of short-chain fatty acids (acetic acid) such as less than 45 mM, less than 44 mM, for example less than 43 mM, less than 42 mM, for example less than 41 mM, less than 40 mM, for example less than 39 mM, in the range of 25 - 45 mM, for example in the range of 28 - 44 mM, in the range of 30 - 43 mM, for example in the range of 32 - 42 mM, in the range of 34 - 41 mM, for example in the range of 36 - 40 mM, in the range of 38 - 39 mM. Preferably, the composition may be a postbiotic.

[0107] The composition according to the present invention may contain one or more live probiotic bacterial strains, or the composition may contain non-live probiotic bacterial strains, or the composition may contain metabolites, lysates, such as fermentation-lysates, obtained from the one or more probiotic bacterial strains, or the composition may contain fractions of the one or more probiotic bacterial strains, or combinations thereof.

[0108] In one embodiment of the present invention, one or more of Lactiplantibacillus plantarum LB990R, deposited with accession number DSM 34494 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited with accession number DSM 34250 on May 5, 2022 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited with accession number DSM 34493 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited with accession number DSM 34492 on January 10, 2023 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited with accession number DSM 34249 on May 5, 2022 by Lactobio A / S; or combinations thereof can be provided as one live cell, dead cell, lysate, fraction, fermentate, fermentate-lysate, metabolite, extract, derivative, analog, or variant of the strain according to the present invention. Preferably, the composition comprises a combination of live cells or dead cells and metabolites from live cells.

[0109] One or more probiotic bacterial strains can be one or more isolated probiotic bacterial strains.

[0110] The term "isolated" can relate to a probiotic bacterial strain that is isolated from a natural source and fermented as a single microorganism, rather than a wild-type fermentate. Preferably, the isolated probiotic bacterial strain according to the present invention can be produced by fermentation as an individual bacterial strain.

[0111] Preferably, one or more lactic acid bacteria (Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S, etc.); and / or one or more lactic acid bacteria (Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S, etc.); and / or one or more lactic acid bacteria (Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S, etc.); and / or one or more lactic acid bacteria (Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S, etc.); one or more lactic acid bacteria (Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S, etc.); one or more lactic acid bacteria or a combination thereof can be provided as live cells, as non-living cells, or as lysates or fermentate - lysates or postbiotics.

[0112] The composition according to the present invention can be formulated for oral use, topical use, anal use or genital use.

[0113] One embodiment of the present invention relates to a composition comprising a probiotic strain according to the present invention, live / living, for use in treating; alleviating, inhibiting; preventing; and / or precluding the growth of pathogenic fungi.

[0114] More preferably, the present invention may provide a composition as defined herein for use in the treatment, alleviation, suppression; prevention of one or more pathogenic bacterial infections in humans or animals.

[0115] A further embodiment of the invention relates to a composition comprising Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 or combinations thereof for use against mycosis.

[0116] A further embodiment of the present invention relates to a composition comprising Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249 or a combination thereof for use against dermatomycosis (tinea corporis) caused by dermatophytes.

[0117] In a further embodiment of the present invention, Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249; or a composition comprising one or more bacterial strains selected from these combinations, for use in the prevention and / or treatment of fungal diseases in humans or animals.

[0118] In particular, the composition according to the invention comprising one or more bacterial strains selected from Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249 or combinations thereof may be suitable for use in the treatment of cutaneous mycosis on epithelial cells, mucosae, or on (or in) the body of a human or animal.

[0119] In particular, the compositions according to the invention comprising one or more bacterial strains selected from Lactiplantibacillus plantarum, LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum, LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum, LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249; or combinations thereof may be suitable for use in the treatment of fungal infections on epithelial cells, mucosae, or on (or in) the body of a human or animal.

[0120] In one embodiment of the invention, the animal can include, but is not limited to, primates, livestock, sports animals, rodents and pets. More specifically, the animal can include mice, rats, hamsters and guinea pigs; rabbits; dogs; cats; sheep; pigs; piglets; sows; poultry; turkeys; broilers; minks; goats; cows; horses; and non-human primates such as apes and monkeys.

[0121] More preferably, the invention may provide a composition as defined herein for use in preventing the growth of pathogenic microorganisms.

[0122] The "decrease" in growth can be "statistically significant" compared to the growth period in the absence of the bacterial strain of the present invention, and can include a decrease of 10 percent, 20 percent, 30 percent, 40 percent, 50 percent, 60 percent, 70 percent, 80 percent, 85 percent, 90 percent, 95 percent, 99 percent or 100 percent.

[0123] In one embodiment of the present invention, growth inhibition can be determined as a decrease in growth of at least 25%. Preferably, growth inhibition is determined as a decrease in growth of at least 50 percent, more preferably, growth inhibition is determined as a decrease in growth of at least 80 percent, and more preferably, growth inhibition is determined as a decrease in growth of at least 90%.

[0124] "Probiotics" can be defined as living microorganisms that confer a health benefit on the host when administered or ingested in an appropriate amount.

[0125] In a preferred embodiment, the lactic acid bacteria of the present invention can be a living, viable strain.

[0126] The non-living cells according to the present invention can be provided as a lysate of one of the strains according to the present invention, as a fraction, as a fermentate, as a fermentate-lysate, as a metabolite, as an active substance, as an extract, as a derivative, as an analog, or as a variant.

[0127] "Lysozyme solution", "ferment - lysozyme solution", "derivative", "analogue", "fraction", "postbiotic" or "extract" can be obtained from dead or inactivated lactic acid bacteria. These lysozyme solutions, fractions, derivatives, analogues, and extracts preferably have the property of being able to bind or co - aggregate with pathogenic fungi, thereby preventing fungal growth and / or adhesion and / or hyphal formation and / or biofilm formation. The terms "lysozyme solution" as well as "extract" and "postbiotic" refer in particular to a solution or suspension in an aqueous medium of the cells and / or metabolic products of the microorganisms according to the invention, for example, macromolecules such as DNA, RNA, proteins, peptides, lipids, carbohydrates, acids, metabolic products produced by lactic acid bacteria, etc., as well as cell debris. The lysozyme solution preferably contains a cell wall or cell wall component containing a binding receptor. In a preferred embodiment, the cells are paraprobiotics which are non - lysed intact dead cells. Methods for producing lysozyme solutions are well - known to those skilled in the art and include, for example, the use of a French press or enzymatic lysis, the use of a ball mill with glass beads or iron beads. The cells can be disrupted and opened by enzymatic, physical or chemical methods. Examples of enzymatic cell lysis can include individual enzymes and enzyme cocktails, such as proteases, proteinase K, lipases, glycosidases; chemical lysis can be induced by ionophores, surfactants such as SDS, acids or bases; physical methods can also be carried out by using high pressure such as a French press, osmotic pressure, temperature, or by alternately repeating heat and cooling. Furthermore, chemical, physical and enzymatic methods can of course be combined to inactivate lactic acid bacteria. "Extract" can be any of these cell components, metabolic products, postbiotics, ferments or cell fractions.

[0128] A preferred embodiment of the present invention is - Lactiplantibacillus plantarum LB990R, deposited on January 10, 2023 with accession number DSM 34494 by Lactobio A / S; and / or - Lactiplantibacillus plantarum LB681R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34250; and / or - Lacticaseibacillus paracasei LB857R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34493; and / or - Lactiplantibacillus plantarum LB760R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34492; and / or - Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34249 relates to an isolated probiotic bacterial strain selected from

[0129] A further preferred embodiment of the present invention is able to partially or completely inhibit the growth of one or more dermatophytes - Lactiplantibacillus plantarum LB990R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34494; - Lactiplantibacillus plantarum LB681R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34250; - Lacticaseibacillus paracasei LB857R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34493; - Lactiplantibacillus plantarum LB760R, deposited on January 10, 2023 with accession number DSM 34492 by Lactobio A / S; - Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited on May 5, 2022 with accession number DSM 34249 by Lactobio A / S It relates to an isolated probiotic bacterial strain selected from the group consisting of:

[0130] Lactiplantibacillus plantarum LB990R, deposited on January 10, 2023 with accession number DSM 34494 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited on May 5, 2022 with accession number DSM 34250 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited on January 10, 2023 with accession number DSM 34493 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited on January 10, 2023 with accession number DSM 34492 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited on May 5, 2022 with accession number DSM 34249 by Lactobio A / S, a variant thereof, using the deposited strain as a starting material and applying mutagenesis, wherein the resulting variant retains or enhances probiotic and / or antifungal and / or antibacterial properties and / or the ability to inhibit mycosis.

[0131] In one embodiment of the present invention, the composition may not contain, or may substantially not contain, living microorganisms. The composition can contain cell material containing dead cells in the form of a lysate, a fermentate, or a fermentate-lysate. In a further embodiment of the present invention, the composition may contain a supernatant from fermentation and cell material having further functional effects.

[0132] In one embodiment of the present invention, the lactic acid bacteria according to the present invention may be capable of inhibiting the growth of fungi.

[0133] The microorganisms according to the present invention may preferably be in an isolated form, and the term "isolated" particularly means that the lactic acid bacteria are derived from their culture media including their natural media, for example, isolated from nature and / or isolated from other species.

[0134] As used herein, the term "inhibit" or "inhibiting" means the killing of microorganisms such as undesirable microorganisms or the control of the growth of said microorganisms. It includes the inhibition of biofilm formation, which may be the inhibition of the initial attachment of microorganisms or the inhibition of the further formation of biofilms by inhibiting the growth of microorganisms.

[0135] In one embodiment of the present invention, the composition and / or lactic acid bacteria according to the present invention may be able to inhibit the growth of fungi.

[0136] In one embodiment of the present invention, the composition and / or lactic acid bacteria according to the present invention may be able to inhibit mycosis.

[0137] In one embodiment of the present invention, the composition and / or lactic acid bacteria according to the present invention may be able to inhibit dermatophytosis.

[0138] One embodiment of the present invention relates to an isolated bacterial strain selected from Lactiplantibacillus plantarum LB990R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34249 or a combination thereof, wherein the strain exhibits antibacterial activity.

[0139] Antifungal activity can be to normalize a dysfunctional microbiota and / or inhibit abnormal fungal growth and / or inhibit mycosis. In particular, Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S; or an isolated bacterial strain selected from these combinations can inhibit the growth of one or more fungi. More specifically, the isolated strain can inhibit the growth of pathogenic fungi. Even more specifically, the isolated strain can inhibit mycosis. Even more specifically, the isolated strain can inhibit dermatophytosis.

[0140] In a preferred embodiment of the invention, the antifungal activity can be determined as inhibition of fungal spore germination. More specifically, the antifungal activity can be determined as inhibition of hyphal formation (hyphal formation of fungi).

[0141] In this context, inhibition of hyphal formation can be, for example, at least 10% hyphal inhibition, at least 25% hyphal inhibition, at least 50% hyphal inhibition, such as at least 75% hyphal inhibition, at least 85% hyphal inhibition, such as at least 90% hyphal inhibition, at least 95% hyphal inhibition, such as at least 98% hyphal inhibition.

[0142] Antifungal activity can be determined as growth inhibition, and inhibition of fungal growth can be determined as a zone of inhibition when an antifungal probiotic strain is placed on a fungal colony grown on an agar plate.

[0143] Antifungal activity can be determined as growth inhibition, and inhibition of fungal growth can be determined as a zone of inhibition when an antifungal probiotic strain is added into a well punched out of agar having a fungal colony grown on an agar plate.

[0144] The inventors of the present invention have identified and isolated specific strains; surprisingly, they have identified one or more specific activities provided by the specific strains, such as inhibition of fungal growth, and / or treatment of a dysfunctional microbiota, and / or prevention of mycosis, and / or treatment of mycosis. of identified and isolated specific strains; and surprisingly, identified one or more specific activities provided by the specific strains, such as inhibition of fungal growth, and / or treatment of a dysfunctional microbiota, and / or prevention of mycosis, and / or treatment of mycosis.

[0145] The inventors of the present invention identified and isolated specific strains of Lactiplantibacillus plantarum LB990R; Lactiplantibacillus plantarum LB681R; Lacticaseibacillus paracasei LB857R; Lactiplantibacillus plantarum LB760R; Lacticaseibacillus paracasei subsp. paracasei LB555R; and surprisingly identified one or more specific activities provided by the specific strains, such as inhibition of dermatophytosis, and / or treatment of dermatophytosis, and / or prevention of dermatophytosis.

[0146] In one embodiment of the present invention, the composition according to the present invention can be used to inhibit the growth of fungi on a surface. Preferably, the surface can be the surface of a biomaterial and / or the surface of a human or animal.

[0147] In one embodiment of the present invention, the composition according to the present invention can be used to inhibit the growth of fungi on a surface. Preferably, the surface can be the surface of a biomaterial and / or the surface of a plant, food, feed, seed or crop.

[0148] A preferred embodiment of the present invention relates to a method for reducing and / or inhibiting spore germination and / or hyphal formation of fungi on a surface, comprising applying the composition according to the present invention to the surface.

[0149] The surface can be a surface infected with fungi, or the composition of the present invention can be applied to a surface that prevents the surface from being infected with fungi.

[0150] Preferably, the surface can be the skin of a human or animal, and / or the surface can be the surface of a crop, seed, food or feed.

[0151] The prevention of a surface infected with a fungus or the prevention of a surface from being infected with a fungus can preferably be a surface infected with one or more dermatophytes, or can prevent the surface from being infected with one or more dermatophytes.

[0152] In one embodiment of the present invention, the composition according to the present invention can be used to inhibit the growth of fungi on a surface. Preferably, the surface can be a surface in a hospital and / or a surface of a medical device.

[0153] In a further embodiment of the present invention, the composition according to the present invention can be used to inhibit the formation of fungi in or on the body of a human or animal.

[0154] In one embodiment of the present invention, the composition (or lactic acid bacteria) according to the present invention can be formulated or used in drugs, pharmaceuticals, foods, feed products, disinfectants, cosmetics, or personal care products.

[0155] The disinfectant according to the present invention can be used to treat a surface to avoid the growth of fungi on the surface. The surface can be a medical or non-medical surface. The medical surface can be a surface in a hospital or a surface of a medical device. The medical device can include a medical device implanted in a human or animal or used inside a human or animal, a rectal insert, a vaginal insert, a dental insert, or a medical device mainly used outside the body of a human or animal.

[0156] One or more of the composition according to the present invention and / or the isolated bacterial strain of the present invention can be used in the treatment of one or more of surgical wounds, pressure ulcers, catheters, stents, heart circulation devices, orthoses, orthosis insertion, otolaryngology, orthopedic and dental orthoses, infections from screws and nails, oral infections, and infections of the oral mucosa and vaginal mucosa, local infections, otitis media, sinusitis, pharyngitis, laryngitis, and pneumonia.

[0157] In embodiments of the present invention, one or more of the compositions according to the present invention and / or the isolated bacterial strains of the present invention may be effective against antibiotic-resistant fungi. In particular, it is effective against antibiotic-resistant infections.

[0158] The use of the composition according to the present invention (or lactic acid bacteria) as a disinfectant can reduce the risk of fungal infection, in particular the risk of infection by pathogenic fungi, and more specifically the risk of infection by fungi.

[0159] The use of the composition according to the present invention (or lactic acid bacteria) as a disinfectant can reduce the risk of skin fungal infection, in particular the risk of infection by dermatophytes, and more specifically, the risk of infection by genera such as Arthoderma, Lophophyton, Nannizzia, Epidermophyton, Microsporum, Deb ar yomyces, Kluyveromyces and Trichophyton.

[0160] One embodiment of the present invention relates to one or more bacterial strains selected from Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250 on May 5, 2022 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493 on January 10, 2023 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492 on January 10, 2023 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 on May 5, 2022 by Lactobio A / S; or combinations thereof, for use as a medicament.

[0161] In one embodiment of the present invention, a composition of the present invention consisting essentially of one or more bacterial strains selected from Lactiplantibacillus plantarum LB990R, deposited on January 10, 2023 with accession number DSM 34494 by Lactobio A / S; Lactiplantibacillus plantarum LB681R, deposited on May 5, 2022 with accession number DSM 34250 by Lactobio A / S; Lacticaseibacillus paracasei LB857R, deposited on January 10, 2023 with accession number DSM 34493 by Lactobio A / S; Lactiplantibacillus plantarum LB760R, deposited on January 10, 2023 with accession number DSM 34492 by Lactobio A / S; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited on May 5, 2022 with accession number DSM 34249 by Lactobio A / S; or combinations thereof.

[0162] In the context of the present invention, the term "consisting essentially of" relates to a limitation of the scope of the claims to the specified features or steps, and features or steps not recited that do not substantially affect the basic and novel characteristics of the claimed invention.

[0163] A preferred embodiment of the present invention relates to a composition comprising one or more bacterial strains selected from one or more lactic acid bacteria for use in the prevention and / or treatment of mycosis in humans or animals.

[0164] Preferably, the prevention and / or treatment of mycosis in humans or animals may be the prevention and / or treatment of mycosis on or in the skin; the prevention and / or treatment of mycosis in the mouth; and / or the prevention and / or treatment of mycosis in the digestive tract, and / or the prevention and / or treatment of mycosis in the intestine.

[0165] The present invention relates to a composition essentially consisting of one or more bacterial strains selected from one or more of the following strains: Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R deposited under accession number DSM 34249, or a combination of one or more thereof, for use in the prevention and / or treatment of mycosis in humans or animals.

[0166] The present invention may relate to a composition consisting essentially of one or more bacterial strains selected from one or more of the following strains: Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250; Lacticaseibacillus paracasei LB857R, deposited under accession number DSM 34493; Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492; Lacticaseibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249, for use in preventing the growth of fungi for the preservation of food, food products, animal feed, crops or seeds.

[0167] In one embodiment of the invention, the antifungal activity is against at least one of the fungal species causing dermatomycosis.

[0168] In one embodiment of the invention, the antifungal activity may be against dermatophytes, which are fungi classified as the family Arthrodermataceae, a family of fungi containing seven genera of dermatophytes: Epidermophyton, Microsporum, Nannizzia, Trichophyton, Paraphyton, Lophophyton and Arthroderma.

[0169] In one embodiment of the present invention, the antifungal activity can be against at least one of the following fungal genera: Arthroderma, Lophophyton, Nannizzia, Epidermophyton, Microsporum, Deb ar yomyces, Kluyveromyces, Candida, Pichia, and Trichophyton.

[0170] In one embodiment of the present invention, the antifungal activity can be against species of the genus Trichophyton.

[0171] In the context of the present invention, the term "pathogenicity" relates to the nature or characteristics of pathogenic fungi that are poisonous, venomous, or harmful to human or animal life or health.

[0172] The composition according to the present invention can preferably consist essentially of one or more bacterial strains selected from Lactocaseibacillus strains.

[0173] One or more bacterial strains selected from Lactocaseibacillus can preferably include Lactiplantibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249 and / or Lactiplantibacillus paracasei LB857R, deposited under accession number DSM 34493.

[0174] The composition according to the present invention can consist essentially of one or more bacterial strains selected from Lactiplantibacillus strains.

[0175] One or more bacterial strains selected from Lactiplantibacillus are preferably Lactiplantibacillus plantarum LB681R, deposited under the accession number DSM34250; and / or Lactiplantibacillus plantarum LB760R, deposited under the accession number DSM 34492; and / or Lactiplantibacillus plantarum LB990R, which may include deposition under the accession number DSM 34494.

[0176] The composition according to the present invention may consist essentially of one or more bacterial strains selected from Lacticaseibacillus strains and Lactiplantibacillus strains.

[0177] The composition may be formulated for oral use, topical use, intestinal use or genital use.

[0178] Preferably, the composition according to the present invention is formulated for oral or genital use. Even more preferably, the composition according to the present invention is formulated for genital use.

[0179] In one embodiment of the present invention, genital use can be female genital use such as vaginal use, and / or male genital use. Preferably, genital use can be vaginal use.

[0180] During topical use of the composition of the present invention, the composition can be used on the skin, in skin folds, on nails, on fingernails, in eyes, in ears, in hair, on feet, on hands and in mucous membranes such as the intestine, digestive tract or the mouth of humans or animals.

[0181] In one embodiment of the present invention, the composition for topical use can be as described in WO 2020 / 127637 or WO 2020 / 249734.

[0182] In one embodiment of the present invention, the composition can be formulated as a suspension, spray, gel, cream, lotion, powder, capsule, oil, cleaning solution, ovule, vaginal insert, suppository, troche, tablet, microencapsulated product, or in the form of a dietary supplement or food.

[0183] The suspension, spray, gel, cream, lotion, powder, capsule, oil, cleaning solution, ovule, vaginal insert, suppository, troche, tablet, microencapsulated product, dietary supplement or food may contain the composition according to the present invention in the range of 0.1-15% (w / w), such as in the range of 0.25-12% (w / w), for example in the range of 0.50-10% (w / w), in the range of 0.75-8% (w / w), for example in the range of 1-6% (w / w), in the range of 2-5% (w / w), for example in the range of 3-4% (w / w).

[0184] The suspension, spray, gel, cream, lotion, powder, capsule, oil, cleaning solution, ovule, vaginal insert, suppository, troche, tablet, microencapsulated product, dietary supplement or food contains 1×10 4 CFU / g to 5×10 11 CFU / g, in the range of 1×10 5 CFU / g to 1×10 11 CFU / g, for example in the range of 5×10 4 CFU / g to 5×10 10 CFU / g, in the range of 1×10 5 CFU / g to 1×10 10 CFU / g, for example in the range of 5×10 5 CFU / g to 5×10 9 CFU / g, and may contain the composition according to the present invention in this range.

[0185] In one embodiment of the present invention, the composition can be formulated as a cream, particularly a foot cream, and the foot cream can contain the composition according to the present invention in the range of 0.1 to 15% (w / w), for example, in the range of 1 to 8% (w / w), in the range of 3 to 6% (w / w), for example, about 5% (w / w). Preferably, the composition can be provided in the form of a lysate (or fermented lysate).

[0186] In one embodiment of the present invention, the composition can be formulated as a cream, particularly a foot cream, and the foot cream can contain the composition according to the present invention in the range of 1×10 4 CFU / g to 5×10 11 CFU / g, for example, in the range of 1×10 5 CFU / g to 1×10 11 CFU / g, for example, in the range of 5×10 4 CFU / g to 5×10 10 CFU / g, for example, in the range of 1×10 5 CFU / g to 1×10 10 CFU / g, for example, in the range of 5×10 5 CFU / g to 5×10 9 CFU / g. Preferably, the composition can be provided in the form of a lysate (or fermented lysate).

[0187] In a further embodiment of the present invention, the composition can be formulated as an oil, particularly a foot oil and / or a nail oil, and the oil can contain the composition according to the present invention in the range of 0.1 to 15% (w / w), for example, in the range of 0.5 to 8% (w / w), in the range of 0.75 to 6% (w / w), for example, in the range of 1 to 4% (w / w), in the range of 1.5 to 2.5% (w / w), for example, about 2% (w / w). Preferably, the composition can be provided in the form of live freeze-dried cells.

[0188] In a further embodiment of the present invention, the composition can be formulated as an oil, particularly a foot oil and / or a nail oil, and the oil can contain the composition according to the present invention in the range of 1×10 4 CFU / g to 5×10 11 CFU / g, for example, in the range of 1×10 5CFU / g to 1×10 11 For example, in the range of 5×10 4 CFU / g to 5×10 10 In the range of 1×10 5 CFU / g to 1×10 10 For example, in the range of 5×10 5 CFU / g to 5×10 9 The composition according to the invention in the range of CFU / g may be included. Preferably, the composition may be provided in the form of live freeze-dried cells.

[0189] In a further embodiment of the invention, the composition may be formulated as a powder, in particular as a foot powder and / or a skin or scalp powder, and the powder may contain the composition according to the invention in the range of 0.1 to 15% (w / w), in the range of 0.2 to 10% (w / w), for example in the range of 0.3 to 8% (w / w), in the range of 0.5 to 6% (w / w), for example in the range of 0.6 to 4% (w / w), in the range of 0.75 to 2.5% (w / w), for example about 1% (w / w) or about 2% (w / w). Preferably, the composition may be provided in the form of live freeze-dried cells.

[0190] In a further embodiment of the invention, the composition may be formulated as a powder, in particular as a foot powder and / or a skin or scalp powder, and the powder may contain 4 1×10 11 CFU / g to 5×10 5 1×10 11 CFU / g to 1×10 4 For example, in the range of 5×10 10 CFU / g to 5×10 5 1×10 10 CFU / g to 1×10 5 For example, in the range of 5×10 9 CFU / g to 5×10

[0191] Preferred embodiments of the present invention relate to food or feed ingredients, or food or feed products, comprising the composition according to the present invention.

[0192] Preferred embodiments of the present invention relate to personal hygiene products comprising the composition according to the present invention.

[0193] Preferred embodiments of the present invention relate to cosmetics comprising the composition according to the present invention.

[0194] The composition according to the present invention can be supplemented with additional ingredients in order to improve the effect of the composition, or to stabilize the composition, or both.

[0195] In one embodiment of the present invention, the composition further comprises an antifungal agent. This additional antifungal agent can further improve the activity of the composition in the prevention or inhibition of biofilm formation.

[0196] The composition according to the present invention can be formulated as a composition for topical use.

[0197] The composition according to the present invention can be formulated in a medical device. Thus, the composition according to the present invention comprising one or more lactic acid bacteria according to the present invention can be used as a medical device.

[0198] The medical device according to the present invention can be formulated in a suspension, spray, gel, cream, lotion, powder, capsule, ointment, oil, cleaning solution, ovule, vaginal insert, suppository, capsule, troche, tablet, microencapsulated product.

[0199] In a further embodiment of the present invention, the composition further comprises one or more prebiotics. The prebiotic can improve the viability of the added lactic acid bacteria and / or the naturally occurring microbiota.

[0200] In one embodiment of the present invention, the composition comprising lactic acid bacteria can further comprise a prebiotic.

[0201] Prebiotics are non-digestible food components that increase 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, potent synbiotics are based on combinations of specific strains of probiotic bacteria and carefully selected prebiotics. Synbiotics can provide important health benefits to humans or animals.

[0202] In embodiments according to the present invention, the composition according to the present invention may comprise a synbiotic composition.

[0203] According to another embodiment of the present invention, a probiotic composition comprising a probiotic microorganism and at least one additional active ingredient may be provided.

[0204] Prebiotics refer to chemical products that induce the growth and / or activity of symbiotic microorganisms (e.g., bacteria and fungi) that contribute to the health of the host. Prebiotics are non-digestible carbohydrates that are not digested or are partially digested by the host and stimulate the growth and / or activity of beneficial bacteria that colonize the host.

[0205] Some oligosaccharides that can be used as prebiotics are fructooligosaccharides (FOS), xylooligosaccharides (XOS), polydextrose, pectin, galactooligosaccharides (GOS), or human milk oligosaccharides (HMO). Additionally, some disaccharides such as lactulose or lactose, and some monosaccharides such as tagatose can also be used as prebiotics.

[0206] Other active ingredients (or other components) can be unlimited in any form.

[0207] Prebiotics can be compounds that can be metabolized by probiotics. Preferably, prebiotics are not digested or are hardly digested by mammals. Prebiotics are well known in the art, and when used in the present invention, there are no particular restrictions on the prebiotic itself.

[0208] In one embodiment of the present invention, at least one prebiotic can be added to the composition. Preferably, the at least one prebiotic can be selected from the following compounds and compositions: indigestible carbohydrates, β-glucan, mannan-oligosaccharide, inulin, oligofructose, human milk oligosaccharide (HMO), galactooligosaccharide (GOS), lactulose, lactosucrose, galactotriose, fructooligosaccharide (FOS), cellobiose, cellodextrin, cyclodextrin, maltitol, lactitol, glycosyl sucrose. Optionally, mannan oligosaccharide and / or inulin may be preferred in some cases.

[0209] HMO 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.

[0210] D- and L-fucose are thought to enhance the natural defense of the skin, stimulate epidermal immune defense, and / or prevent and / or treat skin autoimmune diseases. Thus, in one embodiment of the present invention, the composition may contain D- and / or L-fucose.

[0211] In a further embodiment of the present invention, the composition contains L-fucose at a concentration in the range of 1 mM to 1000 mM, in the range of 10 mM to 500 mM, for example, in the range of 25 mM to 250 mM.

[0212] The composition according to the present invention may further contain at least one active ingredient.

[0213] In one embodiment of the present invention, the composition may contain at least one additional probiotic microorganism selected from the group consisting of bacteria, yeast or mold.

[0214] The term "proliferation" as used in the present invention may relate to the ability of a cell to grow and divide into two or more identical cells.

[0215] The term "living" or "alive" as used herein relates to microorganisms that are not dead and can have an active metabolism.

[0216] The term "microbiota" as used herein relates to the communities of commensal, mutualistic and pathogenic microorganisms found in and on all multicellular organisms. The microbiota includes bacteria, archaea, protists, fungi, yeast, viruses and phages.

[0217] The term "microbiota dysfunction" as used herein relates to a state in which the microbiota functions incorrectly or is completely prevented from functioning. Unless otherwise specified, microbiota dysfunction in the present invention includes abnormal proliferation or increased growth of pathogenic microorganisms that result in a dysfunctional microbiota. An example of a dysfunctional microbiota is an increase in Malassezia furfur that causes dandruff.

[0218] As used herein, the term "probiotic microorganism" relates to living microorganisms that are intended to have health benefits when ingested or applied to a host. Examples of suitable probiotic microorganisms include yeasts such as Saccharomyces, Debaromyces, Candida, Pichia, and Torulopsis; molds such as Aspergillus, Rhizopus, Mucor, Penicillium, and Torulopsis; and bacteria such as the genera Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus, Cutibacterium, Lactiplantibacillus, and Lactobacillus.

[0219] The most commonly used probiotics are strains of lactic acid bacteria (LAB). The term "lactic acid bacteria" includes species of the families Lactobacillaceae, Aerococcaceae, Bifidobacteriaceae, Carnobacteriaceae, Enterococcaceae, Leuconostocaceae, and Streptococcaceae. These are considered to be non-pathogenic and are generally used as probiotic bacteria to improve the gastrointestinal microbiota and in the treatment of gastrointestinal symptoms.

[0220] Lactic acid bacteria are preferably of the genus Lactobacillus, Lactiplantibacillus iGenus Bacillus, genus Holzapfelia, genus Amylolactobacillus, genus Bombilactobacillus, genus Companilactobacillus, genus Lapidilactobacillus, genus Agrilactobacillus, genus Schleiferilactobacillus, genus Loigolactobacillus, genus Lacticaseibacillus, genus Latilactobacillus, genus Dellaglioa, genus Liquorilactobacillus, genus Ligilactobacillus, genus Furfurilactobacillus, genus Paucilactobacillus, genus Limosilactobacillus, genus Fructilactobacillus, genus Acetilactobacillus, genus Apilactobacillus, genus Levilactobacillus, genus Secundilactobacillus, genus Lentilactobacillus, genus Leuconostoc, genus Bifidobacterium, genus Pediococcus, genus Lactococcus, genus Streptococcus, genus Aerococcus, genus Carnobacterium, genus Enterococcus, genus Oenococcus, genus Sporolactobacillus, genus Tetragenococcus,It is selected from the genera Vagococcus and Weissella.

[0221] Preferred lactic acid bacteria are, in particular, bacteria. The bacteria are preferably Lactococcus lactis, Lacticaseibacillus rhamnosus, Lactiplantibacillus plantarum, Lactobacillus helveticus, Lactobacillus jensenii, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus amylovorus, Lactobacillus amylolyticus, Lactobacillus alimentarius, Lactobacillus aviaries, Lactobacillus delbrueckii, Lactobacillus diolivorans, Lactobacillus farciminis, Lactobacillus gallinarum, Lacticaseibacillus casei, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus hilgardii, Lactobacillus kefiranofaciens, Lactobacillus kefiri, Lactobacillus mucosae (Lactobacillusmucosae), Lactobacillus panis, Lactiplantibacillus paraplantarum, Lactobacillus pontis, Latilactobacillus sakei, Lactobacillus saliverius, Lactobacillus sanfra nciscensis), Lacticaseibacillus paracasei, Lactobacillus pentosus, Lactobacillus cellobiosus, Lactobacillus collinoides, Lactobacillus coryniformis, Lactobacillus curvatus, Levilactobacillus brevis, Lactobacillus buchneri, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus ingluviei, Weissella viridescens, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis, Carnobacterium divergens, Corynebacterium glutamicum, Leuconostoc citreum, Leuconostoc lactis, Leuconostoc mesenteroidesmesenteroides), Leuconostoc pseudomesenteroides, Oenococcus oeni, Pasteuria nishizawae, Pediococcus acidilactici, Pediococcus dextrinicus, Pediococcus parvulus, Pediococcus pentosaceus, Pro pi ionibacterium freudenreichii), Pro p ionibacterium freudenreichii), Pro pi ionibacterium acidipro pio nic(Pro p ionibacterium acidiprop ioselected from the group consisting of Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus, Bacillus amyloliquefaciens, Bacillus atrophaeus, Bacillus clausii, Bacillus coagulans, Bacillus flexus, Bacillus fusiformis, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus mojavensis, Bacillus pumilus, Bacillus smithii, Bacillus subtilis, Bacillus vallismortis, Geobacillus stearothermophilus or variants thereof.

[0222] In one preferred embodiment of the present invention, the composition comprises Lactiplantibacillus plantarum LB356R (DSM 33094), Lactiplantibacillus plantarum LB244R (DSM 32996), Weissella viridescens LB10G (DSM 32906), Lacticaseibacillus paracasei LB113R (DSM 32907), Lacticaseibacillus paracasei LB116R (DSM 32908), Levilactobacillus brevis LB152G (DSM 32995), Lacticaseibacillus paracasei LB28R (DSM 32994), Enterococcus faecium LB276R (DSM 32997), Leuconostoc mesenteriodes LB349R (DSM 33093), Lactiplantibacillus plantarum LB316R (DSM 33091), Lactiplantibacillus plantarum LB312R (DSM 33098), Pediococcus pentosaceus LB606R (DSM 33730), Lactiplantibacillus plantarum LB679R (DSM 33731), Lactobacillus crispatus LB714R (DSM 33732), Lactobacillus gasseri LB905R (DSM 34094), Lactobacillus crispatus LB912R (DSMat least one additional strain selected from the group consisting of Lactobacillus jensenii LB918R (DSM 34095), Lactobacillus jensenii LB918R (DSM 34096), Lactobacillus crispatus LB919R (DSM 34097); and / or any combination thereof or mutant strains thereof and / or cell lysates and / or soluble metabolites of any of these probiotic strains.

[0223] The number of microorganisms is measured as colony forming units (CFU) per ml or per gram.

[0224] The microorganisms according to the invention can preferably be in isolated or purified form, and the term "isolated" particularly means that the lactic acid bacteria are derived from their culture media, including for example their natural media. The terms "isolated" and "purified" are not necessarily limited to absolute purity.

[0225] In one embodiment of the invention, the probiotic strain can be used as a living isolated microorganism in a stabilized form. Suitable methods for stabilization are known to those skilled in the art and include freeze drying or lyophilization with different cryoprotectants.

[0226] In a further embodiment of the invention, the strain can be used as a living isolated strain.

[0227] The term "postbiotic" refers to compounds, metabolites or cellular substances secreted or released from probiotic microorganisms that provide health benefits when applied to a host. Postbiotic compositions are characterized by having health benefits without the presence of live microorganisms.

[0228] As used herein, the term "postbiotic fraction of a probiotic microorganism" discloses a fermented composition of a probiotic microorganism that is substantially free of live microorganisms. The composition can include cellular material including dead cells.

[0229] In one embodiment of the invention, the probiotic strain can be used as a live isolated microorganism in a stabilized form. Suitable methods for stabilization are known to those skilled in the art and include freeze-drying, spray-drying or lyophilization with different cryoprotectants.

[0230] Accordingly, the invention further provides a method for the therapeutic or prophylactic control of phytopathogenic fungi of crops, characterized in that an effective (agronomically effective) and non-phytotoxic amount of the antifungal composition according to the invention is applied to seeds, the soil in which the plants grow or can grow, the leaves and / or fruits of the plants or the seeds of the plants.

[0231] The compositions according to the invention are advantageous for controlling fungal diseases of a number of crops, such as cereals, vegetables, solanaceous plants, market garden crops, grapes, fruits in general, and in particular powdery mildew, Septoria disease, Pythium species of these crops.

[0232] The compositions of the invention generally inhibit the growth of a number of fungi that are particularly harmful to crops, especially to grapes, and more specifically powdery mildew of grapevines.

[0233] These compositions include not only compositions that are ready to be applied to treat crops or seeds by means of a suitable device such as a spraying device, but also commercially available concentrated compositions that need to be diluted with a carrier or mixed with a carrier before application to the crops.

[0234] The invention provides a biological method for controlling a wide variety of phytopathogenic diseases of crops.

[0235] Accordingly, the present invention provides a method for the therapeutic or prophylactic control of phytopathogenic diseases of crops, which comprises treating the crops with an effective and non-phytotoxic amount of the composition of the present invention (e.g., by application or administration). The expression "treatment of crops" means, for example, the application or administration of the above-mentioned antifungal composition to the aerial parts of the crops or to the soil in which the crops are growing, which is invaded or potentially invaded by phytopathogens such as powdery mildew.

[0236] It means the application or administration of the above-mentioned antifungal composition to the soil in which phytopathogenic diseases such as Septoria leaf blotch are prevalent or likely to prevail. The expression "treatment of crops" also means, for example, the treatment of reproduction products of crops such as seeds or tubers.

[0237] The compositions described below are generally used for application to growing plants or to the areas where the crops are grown, or for seed coating or film coating of seeds.

[0238] The expression "applied to the plants to be treated" is understood herein to mean that the antifungal composition of the present invention can be applied by various treatment processes such as spraying the liquid containing the composition onto the aerial parts of the plants, sprinkling, incorporating granules or powders into the soil, watering around the plants, and film coating or forming on the seeds of the plants using a broth or carrier containing the composition.

[0239] In addition to the control of fungal diseases and crop protection, further embodiments of the present invention are the inhibition or growth control of fungi that cause food spoilage or other concerns in the food processing industry. In a preferred embodiment of the present invention, lactic acid bacteria and / or fermented lysate can control the growth of "spoilage" fungi. As used herein, the term "spoilage fungi" refers to any food or feed fungi that cause disease or illness in animals or humans and / or cause food spoilage. The term spoilage fungi is understood to include fungi that have the ability to grow and / or spoil edible substances, thereby causing spoilage, disease or illness after ingestion by mammals, as well as fungi that produce toxins that cause disease or illness. The growth of spoilage fungi on food or feed products can cause serious illness and can be lethal, as evidenced by the number of deaths caused by food poisoning. The term "undesirable fungi" as used herein refers to both spoilage and pathogenic fungi.

[0240] The term "edible substance" or "edible product" refers to a substance or product that is safe for oral ingestion by humans or animals and includes food and feed products as well as ingredients for food or feed products.

[0241] The term "food" as used herein refers to any food that is prone to spoilage as a result of the growth and proliferation of microorganisms on the surface of the food. Such foods include, but are not limited to, meat, dairy products, vegetables, fruits and grains.

[0242] As used herein, the term "meat" refers to any raw meat product, processed meat or meat by-products derived from animals of the kingdom Animalia, including but not limited to meat from cows, sheep, pigs, poultry, frogs, fish and crustacean seafood, which are consumed by humans or animals. Thus, one of the main uses of the present invention relates to meat processed in the butchering of mammals in meat processing facilities, but it should be clearly understood that the present invention has applications in the processing of other edible meat products including fish, poultry and seafood as well as cultured meat. Dairy products are also included, including hard cheeses, yogurts, and dairy products where the growth of fungi is not desired. Furthermore, it is contemplated that the method also has applications related to the preservation of non-animal foods such as fruits, vegetables, bread, bread products, cakes, grains and grain-based products, which are susceptible to spoilage by microorganisms.

[0243] In a preferred embodiment, the meat is cultured meat. In addition to "cultured meat", the terms healthy meat, slaughter-free meat, in vitro meat, vat-grown meat, lab-grown meat, cell-based meat, clean meat, cultivated meat and synthetic meat are all used by various channels to describe such products. Typically, cultured meat is grown from stem cells. Some of the first descriptions of cultured meat are described in U.S. Patent No. 6,835,390, which relates to the production of meat made by tissue engineering for human consumption, growing muscle and fat cells together to produce foods such as beef, poultry and fish.

[0244] As in all descriptions of ranges given in the present invention, characterized by terms such as "about" or "substantially", it is not necessary for an exact numerical range to be indicated by expressions such as "about" or "approximately" or "substantially", and instead it will be apparent to those skilled in the art that even a slight deviation up or down from the indicated number is still within the scope of the present invention.

[0245] The composition according to the present invention may comprise at least one bacterial strain in combination with at least one further probiotic microorganism, and the at least one further probiotic microorganism may be selected from strains selected from the genus Lactobacillus, but is not limited thereto. Deposit of Biological Materials

[0246] The lactic acid bacteria according to the present invention specifically include microorganisms or analogs, fragments, derivatives, fermentates, lysates, fermentate-lysates, mutants or combinations obtained from microorganisms. The microorganisms were deposited with the German Collection for Microorganisms and Cell on May 5, 2022. Lactiplantibacillus plantarum subsp. plantarum LB555R, deposited under accession number DSM 34249, and Lactiplantibacillus plantarum LB681R, deposited under accession number DSM 34250, and Lactiplantibacillus plantarum LB760R, deposited under accession number DSM 34492, Lactiplantibacillus plantarum LB990R, deposited under accession number DSM 34494, and Lactiplantibacillus paracasei LB857R, deposited under accession number DSM 34493.

[0247] It should be noted that the embodiments and features described in one context of the aspects of the present invention are also applicable to other aspects of the present invention.

[0248] All patents and non-patent references cited in this application are hereby incorporated by reference in their entirety.

[0249] Here, in the following non-limiting examples, the present invention will be described in more detail.

Example

[0250] Example 1: Strain screening and identification

[0251] For the identification and selection of microorganisms according to the present invention, a new collection of isolated lactic acid bacteria (LAB) strains was established. To isolate at least 1500 new lactic acid strains, samples of different origins such as homemade sauerkraut, kefir, kimchi, fermented foods, and healthy human donor samples (vagina, oral cavity, anus, skin, human milk, baby diapers) were collected. Samples were collected on Man Rogosa Sharp (MRS, Sigma-Aldrich) broth and anaerobically cultured at 37 °C overnight or until colony formation on agar. New isolates were seeded and subcultured until isolated colonies were obtained. For future use, the isolated colonies were stored at -80 °C in MRS broth with 25% glycerol. Strains were identified using the 16S rRNA Sanger sequencing standard method.

[0252] Example 2: Inhibition of M. furfur DSMZ 6170, M. restricta CBS 7877, and M. globosa CBS 7874

[0253] Inhibition was evaluated using a modified spot-on lawn method from Zhang P. et al. (2015) Inter-strain interactions between bacteria isolated from vacuum-packaged refrigerated beef. Appl Environ Microbiol 81:2753-2761. doi:10.1128 / AEM.03933-14 and Arena, M. P. 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. doi:10.3389 / fmicb.2016.00464.

[0254] The target strain Malassezia furfur DSMZ 6170 was obtained from the Leibniz Institute DSMZ. The target strains M. globosa CBS 7874 and M. restricta CBS 7877 were obtained from the Westerdijk Fungal Biodiversity Institute CBS. The target strains were grown on modified Leeming Notmann (mLN) (ATCC medium number 2737 Leeming & Notman agar modified) agar, and a single colony was inoculated into 10 mL of mLN Broth. The target strain cultures were used to prepare fungal colonies and dried. The bacterial strains isolated from Example 1 were cultured from storage in 10 mL of MRS broth. Subsequently, the overnight cultures of LAB were placed as spots on the plates and incubated. M. furfur was grown at 30 °C for approximately 10 days, M. restricta was grown at 30 °C for approximately 14 days, and M. globosa was grown at 33 °C for approximately 14 days. The inhibition zone was measured in millimeters from the edge of the LAB colony to the start of visible Malassezia growth (the clearing zone). All plates were performed in technical duplicates and repeated at different times.

[0255] LAB was screened against M. furfur DSMZ 6170, of which 84 showed moderate inhibition and 18 showed high inhibition. Two strains were selected from the 18 strains and subsequently tested using the spot-on-colony method against M. restricta CBS 7877 and M. globosa CBS 7874. Both strains were able to inhibit M. restricta CBS 7877 and M. globosa CBS 7874.

[0256] Table 1 As a target, a spot assay using Malassezia species, the inhibition zone / clearing zone existing between the outer edge of the bacterial colony and the visible growth of Malassezia species, measured in millimeters as the average of two sets.

Table 1

[0257] Example 3: A Trichophyton species spot-on agar assay was performed, and the antifungal activity against Trichophyton species was determined as described in Example 2.

[0258] The Trichophyton rubrum CBS 189.69 nail isolate was obtained from the Westerdijk Fungal Biodiversity Institute of the Royal Netherlands Academy of Arts and Sciences.

[0259] The Trichophyton rubrum CBS 392.58 (neotype specimen of Epidermophyto rubrum Castell) skin (foot) isolate was obtained from the Westerdijk Fungal Biodiversity Institute of the Royal Netherlands Academy of Arts and Sciences.

[0260] Trichophyton species were grown on Sabouraud maltose agar at 24°C for 2 - 3 weeks.

[0261] Table 2 Spot assay using Trichophyton species. As a target, the inhibition zone / clearing zone present between the outer edge of the bacterial colony and the visible growth of Trichophyton species, measured in millimeters as the average of a pair.

Table 2

[0262] Example 4: Inhibition by cell-free supernatant solution

[0263] The growth of Malassezia species (two strains described in Example 2) with and without cell-free supernatant (CFS) of LAB from Example 2 was monitored using oCelloScope (BioSense Solutions ApS, Denmark).

[0264] 5 - 10 mL of LAB cell culture was centrifuged (2,700×g, 10 minutes) and sterile filtered using a 0.2 μm syringe filter. Samples were taken from the LAB culture after 24 - 72 hours.

[0265] 1:2 dilutions were prepared from 10 mL of Malassezia species culture. From each of these dilutions, 100 μL was mixed with 100 μL of CSF or its dilutions using 100, 75, 50, 25, 10, and 0% CSF in a 96-well microtiter plate. The volume was adjusted using MRS broth. As a positive control for inhibition, Malassezia species was treated with fluconazole (64 μg / mL). All loading assays were performed in technical triplicates and repeated at separate times. The plates were incubated at 37 °C in oCelloScope. Live images were taken every two and a half hours until 48 hours, and the growth curves were plotted in Excel and calculated as the average of triplicates.

[0266] After the experiment, the plates were visually inspected and the raw images were examined visually. The images were analyzed using UniExplorer PC software.

Brief Description of the Drawings

[0267] The results are shown in Figures 1 - 3, and it was demonstrated that the cell - free supernatants obtained from two isolated strains had a strong and significant inhibition compared to the composition containing the antifungal compound fluconazole (at a concentration of 64 μg / ml) and compared to the non - treated fungal solution when using compositions containing (i) Lactiplantibacillus paracasei subsp. paracasei LB555R, deposited under accession number DSM 34249; (ii) Lactiplantibacillus plantarum, LB681R, deposited under accession number DSM34250.

Figure 1

Figure 2

Figure 3

[0268] The two selected strains were identified in the screening as lactic acid bacteria capable of inhibiting the fungal growth of at least two pathogenic fungi, and the growth inhibition was significantly better than that observed against the known antifungal antibiotic fluconazole.

[0269] Example 5: Analysis of short - chain fatty acid production (SCFA)

[0270] The sample analysis was performed as follows. Using hydrochloric acid, the cell-free supernatant from the strain grown in MRS medium at 37 °C for 24 hours was acidified, and a deuterium-labeled internal standard substance was added. The analysis was carried out using a highly polar column (Zebron™ ZB-FFAP, GC Cap.Column 30m×0.25mm×0.25μm) installed in a GC (7890B, Agilent) connected to a quadrupole detector (5977B, Agilent). The system was controlled by ChemStation (Agilent). After converting the raw data into the netCDF format using Chemstation (Agilent), the data was imported and processed in Matlab R2014b (Mathworks, Inc.) using the PARADISe software described by Johnsen et al. (DOI: 10.1016 / j.chroma.2017.04.052). The SCFA method is a GC-MS method that targets particularly short-chain fatty acids using a highly polar column and standard substances. The concentration of short-chain fatty acid (acetic acid) in the supernatant from each strain was determined. TIFF2025518237000026.tif17130

[0271] Example 6: Well diffusion assay

[0272] An overnight culture of LAB was prepared. LAB was grown in MRS broth, and the fungi grown as described in the above examples for the spots on the bacterial lawn were tested. Wells were made in the plate, and 50 μL of the LAB culture was transferred to the wells. After incubation until visible fungal growth, the inhibition zone around the wells was examined.

[0273] Table 3: Inhibition zones in mm are measured for spots on the bacterial lawn and diffusion assays.

Table 3

[0274] Example 7: Inhibition of Trichophyton rubrum CBS 189.69 and T. rubrum CBS 392.58

[0275] Inhibition was evaluated using a spot-on-the-inoculum method modified from Zhang P. et al (2015) Appl Environ Microbiol 81:2753-2761. Doi:10.1128 / AEM.03933-14 and Arena, M.P. et al. (2016) Frontiers in Microbiology 7(APR):1-10. Doi:https: / / doi.org / 10.3389 / fmicb.2016.00464.

[0276] Replacing the Sabouraud maltose agar used in Examples 3 and 6 with Mueller Hinton agar allowed for a more standardized and uniform Trichophyton inoculum and smaller inhibition zones. This ensures that when more than one spot is formed on the inoculum, the inhibition zones do not interfere or overlap, providing a more accurate comparison of the inhibition zones.

[0277] The target strains T. rubrum CBS 392.58 and T. rubrum CBS 189.69 were obtained from the Westerdijk Fungal Biodiversity Institute CBS. The target strains were grown in malt extract agar (MEA) or malt extract broth (MEB). Spores were harvested from the target strains grown on MEA using a 50% glycerol solution. This spore solution was used to prepare the target strain inoculum on Mueller-Hinton (MH) agar.

[0278] The bacterial strains from Example 1 were cultured from storage in 1 mL of MRS broth. Subsequently, an overnight culture of the LAB was placed as a spot on the plate and incubated at 24 °C for approximately 14 days. The inhibition zone was measured in millimeters from the edge of the LAB colony to the onset of visible T. rubrum growth (the clearing zone). All plates were performed in technical duplicates and repeated at different times (biological replicates).

[0279] 226 pre-selected LAB were screened against T. rubrum CBS 392.58 and T. rubrum CBS 189.69, of which approximately 70 showed varying degrees of inhibition. Six strains were identified as having significant growth inhibitory activity against T. rubrum.

[0280] Table 4 Spot assay on MH agar using T. rubrum species, inhibition zone / clearing zone present between the outer edge of the bacterial colony and the visible growth of two strains of the T. rubrum species, measured in millimeters as the average of four biological and two technical replicates.

Table 4

[0281] Example 8: Inhibition of fungal spore germination by cell-free supernatant solution

[0282] The fungal spore germination of T. rubrum CBS 392.58 was monitored with or without the CFS of the LAB from Example 2 using an oCelloScope (BioSense Solutions ApS (Hirsemarken 1, DK-3520 Farum)).

[0283] 5 - 10 mL of LAB cell culture was centrifuged (2,700×g, 10 minutes) and sterile filtered using a 0.2 μm syringe filter. Samples were taken from the LAB culture after 72 hours.

[0284] A 50% glycerol spore solution collected from an MEA plate containing T. rubrum CBS 392.58 was diluted with MEB (20 μL spore solution in 105 μL MEB). Subsequently, this was treated with different concentrations of cell - free supernatant (CFS) diluted with MRS broth (75 μL, 50 μL, 25 μL, 10 μL, 5 μL).

[0285] 226 pre - selected LAB were screened against T. rubrum CBS 392.58 and T. rubrum CBS 189.69, and approximately 70 of them showed varying degrees of inhibition. Six strains were identified as having significant inhibition of fungal spore germination and hyphal formation of the T. rubrum strains.

[0286] All loading assays were performed in technical triplicates and repeated at separate times. Plates were incubated at 24 °C in an oCelloScope. Live images were taken every 2 hours up to a maximum of approximately 100 hours. After the experiment, plates were visually inspected and live images were examined visually. Inhibition was classified according to the following scale (calculated as the average of triplicates): No germination: 0 (0 germinated spores) Virtually no germination: 1 (1 - 5 germinated spores out of 30 - 50 spores) Germination: 2 (5 - 10 germinated spores out of 30 - 50 spores) High germination: 3 (more than 10 germinated spores out of 30 - 50 spores) Complete germination: 4 (all spores have germinated by more than 95%) TIFF2025518237000029.tif31128

[0287] Example 9: Different formulations

[0288] A) Ingredients provided in foot cream - %(w / w): Aqua 45% Cetearyl alcohol 14% Sodium cetearyl sulfate 10% Prunus amygdalus dulcis oil 6% Prunus armeniaca kernel oil 6% Glycerin 5% Butyrospermum parkii 5% LB990R fermented lysate 5% Vitis vinifera 2% Hydroxypropyltrimonium chloride starch 1% Phenoxyethanol 1%

[0289] B) Ingredients provided in foot oil - %(w / w): Prunus amygdalus dulcis oil 50% Prunus armeniaca kernel oil 47% Lactiplantibacillus plantarum LB990R freeze - dried (at a concentration of 3×10 11 CF / g of viable freeze - dried concentration) 2% Tocopherol 1%

[0290] C) Ingredients provided in nail oil - %(w / w): Castor oil 35% Prunus amygdalus dulcis oil 32% Prunus armeniaca kernel oil 30% Lactiplantibacillus plantarum LB681R freeze - dried (4×1011 Viable freeze-dried concentration of CFU / g) 1% Lactiplantibacillus plantarum LB990R freeze-dried (1×10 11 Viable freeze-dried concentration of CFU / g) 1% Tocopherol 1%

[0291] D) Components provided in foot powder - %(w / w): Components: Talc 50% Rice starch 45% Lacticaseibacillus paracasei LB857R freeze-dried (viable freeze-dried) 2%

[0292] E) Components provided in skin or scalp powder - %(w / w): Corn starch 95% Magnesium stearate 4% Lactiplantibacillus plantarum LB760R freeze-dried (2×10 10 Viable freeze-dried concentration of CFU / g) 1%

[0293] Using the colony spot test described in Example 7, all formulations were tested for antifungal activity against Trichophyton rubrum CBS 189.69.

[0294] For each formulation, 4 spots were made and the formulation was spotted directly onto the colony. Antifungal activity was determined if fungal growth did not occur in the spotted area and at least a 1 mm clearing zone was visible around the spotted formulation. The powder formulation was suspended in sterile saline (0.9% W / W) and 10 μL was spotted.

[0295] The results obtained from the tests showed that the antifungal activity of these strains was maintained in the formulations, that all formulations did not result in fungal growth in the spotted areas, and that the clearance zones around the spots were determined in mm. TIFF2025518237000030.tif37128

Prior Art Documents

Non-Patent Documents

[0296]

Non-Patent Document 1

Non-Patent Document 2

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Claims

**Claim 1** A composition comprising one or more probiotic bacterial strains, wherein the composition can partially or completely inhibit the growth of one or more dermatophytes. **Claim 2** The composition according to claim 1, wherein the one or more probiotic bacterial strains are one or more isolated probiotic bacterial strains or variants thereof. **Claim 3** The composition according to any one of claims 1 to 2, wherein the composition does not contain plant material and / or fiber material. **Claim 4** The composition according to any one of claims 1 to 3, wherein the one or more dermatophytes are selected from Arthroderma, Lophophyton, Nannizzia, Epidermophyton, Microsporum, Debaryomyces, Kluyveromyces, Pichia, Trichophyton, or combinations thereof. **Claim 5** The composition according to any one of claims 1 to 4, wherein the one or more probiotic bacterial strains can be selected from Lactiplantibacillus plantarum and / or Lactiplantibacillus paracasei. **Claim 6** The one or more probiotic bacterial strains are - Lactiplantibacillus plantarum LB990R, deposited with accession number DSM 34494 on January 10, 2023 by Lactobio A / S; - Lactiplantibacillus plantarum LB681R, deposited with accession number DSM 34250 on May 5, 2022 by Lactobio A / S; - Lactiplantibacillus paracasei LB857R, deposited with accession number DSM 34493 on January 10, 2023 by Lactobio A / S; - Lactiplantibacillus plantarum LB760R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34492; - Lactiplantibacillus paracasei subsp. paracasei LB555R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34249; or - a combination thereof The composition according to any one of claims 1 to 5, selected from one or more of the above.

7. The composition according to any one of claims 1 to 6, wherein the composition comprises one or more live probiotic bacterial strains, or the composition comprises non-live probiotic bacterial strains, or the composition comprises metabolites, lysates, such as fermentation-lysates, obtained from the one or more probiotic bacterial strains, or the composition comprises fractions of the one or more probiotic bacterial strains, or combinations thereof.

8. An antifungal composition comprising one or more probiotic bacterial strains for inhibiting fungal spore germination and / or inhibiting fungal hyphal formation.

9. The antifungal composition according to claim 8, wherein the one or more probiotic bacterial strains are selected from one or more Lactiplantibacillus plantarum, one or more Lactiplantibacillus paracasei, or a combination thereof.

10. The above-mentioned one or more Lactiplantibacillus plantarum are deposited by Lactiplantibacillus plantarum LB990R, Lactobio A / S on January 10, 2023 under accession number DSM 34494; Lactiplantibacillus plantarum LB681R, Lactobio A / S on May 5, 2022 under accession number DSM 34250; Lactiplantibacillus plantarum LB760R, Lactobio A / S on January 10, 2023 under accession number DSM 34492, and are selected from; and / or the above-mentioned one or more Lactiplantibacillus paracasei are deposited by Lactiplantibacillus paracasei LB857R, Lactobio A / S on January 10, 2023 under accession number DSM 34493; Lactiplantibacillus paracasei subsp. paracasei LB555R, Lactobio A / S on May 5, 2022 under accession number DSM 34249; or are selected from combinations thereof, the antifungal composition according to claim 9.

11. The antifungal composition according to any one of claims 8 to 10, wherein the fungus is a dermatophyte.

12. A method for reducing and / or inhibiting spore germination of fungi on a surface and / or hyphal formation of fungi, the method comprising applying the composition according to the present invention to the surface.

13. - Lactiplantibacillus plantarum LB990R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34494; - Lactiplantibacillus plantarum LB681R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34250; - Lactiplantibacillus plantarum LB857R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34493; - Lactiplantibacillus plantarum LB760R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34492; - Lactiplantibacillus plantarum LB555R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34249 An isolated probiotic bacterial strain selected from the group consisting of: **Claim 14** One or more skin filamentous fungi whose growth can be partially or completely inhibited, - Lactiplantibacillus plantarum LB990R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34494; - Lactiplantibacillus plantarum LB681R, deposited by Lactobio A / S on May 5, 2022 under accession number DSM 34250; - Lactiplantibacillus plantarum LB857R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34493; - Lactiplantibacillus plantarum LB760R, deposited by Lactobio A / S on January 10, 2023 under accession number DSM 34492; - An isolated probiotic bacterial strain selected from Lactiaseibacillus paracasei subsp. paracasei LB555R, deposited with accession number DSM 34249 by Lactobio A / S on May 5, 2022 from the isolated probiotic bacterial strains selected.

15. - The treatment or prevention of fungal diseases, preferably the treatment and / or prevention of fungal diseases in humans or animals; - The treatment of dermatophytosis, preferably the treatment and / or prevention of dermatophytosis in humans or animals; and / or The prevention, inhibition and / or treatment of the growth of fungi in crops, seeds, food or feed For use in, Lactiplantibacillus plantarum LB990R, deposited with accession number DSM 34494; Lactiplantibacillus plantarum LB681R, deposited with accession number DSM 34250; Lactiplantibacillus plantarum LB760R, deposited with accession number DSM 34492; Lactiaseibacillus paracasei LB857R, deposited with accession number DSM 34493; Lactiaseibacillus paracasei subsp. paracasei LB555R deposited with accession number DSM 34249; or a composition comprising a combination thereof.

Images