Lacticaseibacillus casei VB179 for inhibiting malassezia, and culture device and use thereof

Abstract

The present invention relates to the technical field of microorganisms. Disclosed in the present invention are Lacticaseibacillus casei VB179 for inhibiting Malassezia, and a culture device and the use thereof. Specifically, disclosed in the present invention is Lacticaseibacillus casei VB179, wherein the Lacticaseibacillus casei VB179 is deposited in the China General Microbiological Culture Collection Center with a deposit number of CGMCC No. 28623 and a deposit date of 12 October 2023. The Lacticaseibacillus casei VB179 of the present invention can significantly inhibit the growth of Malassezia, and can further be used for treating skin diseases caused by the excessive reproduction of Malassezia, such as tinea versicolor, Malassezia folliculitis, seborrheic dermatitis and atopic dermatitis.

Description

A type of Lactobacillus casei VB179 that inhibits Malassezia, its culture device and application Technical Field

[0001] This invention belongs to the field of microbial technology, and in particular relates to a type of Lactobacillus casei VB179 that can significantly inhibit Malassezia, its culture device, and its application. Background Technology

[0002] Malassezia is an opportunistic pathogen with a lipophilic affinity for lipids. It primarily grows in the stratum corneum of human skin and is associated with sebaceous glands containing triglycerides and free fatty acids on the skin surface. Its density decreases with age due to a reduction in lipid content. It can be detected in human skin and hair samples. Numerous studies have shown it to be an opportunistic pathogen, mostly harmless. However, under certain internal and external conditions, Malassezia can cause tinea versicolor, pityriasis folliculitis, seborrheic dermatitis and dandruff, atopic dermatitis, and psoriasis. With the expansion of research, studies have also confirmed its association with onychomycosis and systemic infections.

[0003] Therefore, finding a drug that can inhibit the excessive proliferation of Malassezia is of utmost urgency. Ketoconazole, itraconazole, and other azole antifungal drugs were once favored by patients due to their significant efficacy, but they often cause side effects such as skin redness and swelling, and swelling of some hair follicles. Furthermore, over time, it has been found that these drugs have significant side effects and a high relapse rate, necessitating the development of novel Malassezia inhibitors.

[0004] Therefore, this invention screened a biopharmaceutical that inhibits the growth of Malassezia, providing a new approach for developing therapeutic topical agents and shampoos for treating dandruff. Summary of the Invention

[0005] This invention aims to at least partially address one of the technical problems existing in the prior art. To this end, this invention provides for the first time a *Lactaseibacillus casei* VB179 derived from yogurt, which significantly inhibits the growth of *Malassezia*, and can thus be used to treat skin diseases such as tinea versicolor, Malassezia folliculitis, seborrheic dermatitis, and characteristic dermatitis caused by the overgrowth of *Malassezia*. Therefore, using the *Lactaseibacillus casei* VB179 of this invention, a *Malassezia* inhibitor can be prepared and used as a drug for the prevention, relief, and treatment of the aforementioned skin diseases, as well as in daily chemical products. Furthermore, the *Lactaseibacillus casei* VB179 of this invention also exhibits broad antibacterial activity, inhibiting various other pathogenic bacteria, including *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, providing potential treatment options and possibilities for further treatment of diseases associated with these pathogenic bacteria.

[0006] Therefore, in a first aspect, the present invention provides a *Lactaseibacillus casei* VB179. According to an embodiment of the present invention, the *Lactaseibacillus casei* VB179 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 28623, deposited on October 12, 2023, at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The *Lactaseibacillus casei* VB179 of the present invention exhibits broad antibacterial activity, significantly inhibiting not only the growth of *Malassezia*, but also various other pathogenic bacteria, including *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, providing potential therapeutic options and possibilities for further treatment of diseases associated with these pathogens.

[0007] In a second aspect, the present invention provides a fermentation broth, bacterial suspension, or bacterial residue. According to embodiments of the present invention, the fermentation broth, bacterial suspension, or bacterial residue includes *Lactobacillus casei* VB179 as described in the first aspect. Therefore, the fermentation broth, bacterial suspension, or bacterial residue of the present invention can also significantly inhibit the growth of *Malassezia*, and thus can be used to treat skin diseases such as tinea versicolor, Malassezia folliculitis, seborrheic dermatitis, and characteristic dermatitis caused by the overgrowth of *Malassezia*.

[0008] In a third aspect, the present invention provides a microbial inoculant. According to embodiments of the present invention, the microbial inoculant comprises *Lactobacillus casei* VB179 as described in the first aspect or fermentation broth, bacterial suspension, or bacterial residue as described in the second aspect. Therefore, the microbial inoculant of the present invention not only significantly inhibits the growth of *Malassezia*, but also exhibits inhibitory effects on a variety of other pathogenic bacteria, including *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, providing potential treatment options and possibilities for further treatment of diseases associated with these pathogenic bacteria.

[0009] In a fourth aspect, the present invention provides a single-dose formulation. According to embodiments of the invention, the single-dose formulation comprises not less than 10 6 The active ingredient is *Lactobacillus casei* VB179, as described in the first aspect of CFU. Therefore, the single-dose formulation of the present invention not only significantly inhibits the growth of *Malassezia*, but also exhibits inhibitory effects on a variety of other pathogenic bacteria, including *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, providing potential therapeutic options and possibilities for further treatment of diseases associated with these pathogens.

[0010] In a fifth aspect of the invention, the invention proposes the use of the *Lactobacillus casei* VB179 described in the first aspect, the fermentation broth, bacterial suspension or bacterial residue described in the second aspect, the microbial agent described in the third aspect, or the single-dose formulation described in the fourth aspect in the preparation of a medicament for the prevention and / or relief and / or treatment of related diseases or symptoms caused by pathogenic bacterial infection.

[0011] In a sixth aspect, the present invention provides a product. According to embodiments of the invention, the product comprises at least one of the following: *Lactobacillus casei* VB179 as described in the first aspect, fermentation broth, bacterial suspension, or bacterial residue as described in the second aspect, microbial agent as described in the third aspect, and single-dose formulation as described in the fourth aspect. Thus, the product of the present invention can be used to inhibit the growth of *Malassezia*, *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, thereby preventing and / or alleviating and / or treating diseases or symptoms associated with these pathogens.

[0012] In a seventh aspect, the present invention provides a method for inhibiting the growth of pathogenic bacteria in vitro. According to an embodiment of the present invention, the method comprises: co-culturing at least one of the following: *Lactobacillus casei* VB179 as described in the first aspect, the fermentation broth, bacterial suspension, or bacterial residue as described in the second aspect, the microbial agent as described in the third aspect, and the single-dose formulation as described in the fourth aspect, with a sample containing pathogenic bacteria. Using the method of the present invention, the growth of pathogenic bacteria can be effectively inhibited, thereby effectively controlling the infections and diseases they cause, providing an efficient and reliable approach for the treatment and prevention of diseases.

[0013] In an eighth aspect of the invention, the invention provides a culture or production apparatus for Lactobacillus casei VB179 as described in the first aspect, fermentation broth, bacterial suspension or bacterial residue as described in the second aspect, microbial agent as described in the third aspect, or single-dose formulation as described in the fourth aspect.

[0014] In a ninth aspect of the invention, the invention provides the use of the Lactobacillus casei VB179 described in the first aspect, the fermentation broth, bacterial suspension or bacterial residue described in the second aspect, the microbial agent described in the third aspect, or the single-dose formulation described in the fourth aspect in inhibiting the growth of pathogenic bacteria.

[0015] In a tenth aspect of the invention, the invention proposes the use of Lactobacillus casei VB179 as described in the first aspect, fermentation broth, bacterial suspension or bacterial residue as described in the second aspect, microbial agent as described in the third aspect, or single-dose formulation as described in the fourth aspect for the prevention and / or relief and / or treatment of related diseases or symptoms caused by pathogenic bacterial infections.

[0016] In an eleventh aspect of the invention, a method for preventing and / or alleviating and / or treating diseases or symptoms caused by pathogenic bacterial infections is provided. According to an embodiment of the invention, the method comprises administering to a patient at a pharmaceutically acceptable dose of at least one of the following: *Lactobacillus casei* VB179 as described in the first aspect, fermentation broth, bacterial suspension, or bacterial residue as described in the second aspect, microbial agent as described in the third aspect, or single-dose formulation as described in the fourth aspect. Beneficial effects:

[0017] This invention is the first to discover *Lactobacillus casei* VB179, which has an inhibitory effect on the growth of *Malassezia*. Therefore, *Lactobacillus casei* VB179 of this invention can be used to prepare *Malassezia* inhibitors and can be used as drugs and daily chemical products for the prevention, relief, and treatment of skin diseases such as tinea versicolor, Malassezia folliculitis, seborrheic dermatitis, and characteristic dermatitis. Furthermore, *Lactobacillus casei* VB179 of this invention also has inhibitory activity against various pathogenic bacteria, inhibiting the growth of pathogenic bacteria such as *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*. Therefore, it can be used alone or in combination with other probiotics to prevent, relieve, and treat related diseases caused by the aforementioned pathogenic bacteria. Finally, *Lactobacillus casei* VB179 of this invention, as a common probiotic, has high safety, is easy to grow and reproduce, has low production costs, and a simple process. These advantages demonstrate its optimistic prospects in multiple application fields and provide strong support for the development and innovation of related industries.

[0018] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0019] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0020] Figure 1 shows the inhibitory effect of Lactobacillus casei VB179 on Malassezia according to an embodiment of the present invention (negative control: sterile water; positive control: ketoconazole 0.1 mg / L);

[0021] Figure 2 shows the differences in inhibition zones of different Lactobacillus casei VB178, VB179, VB180, and VB217 against Malassezia according to embodiments of the present invention.

[0022] Figure 3 shows the differences in inhibition zones of different *Lactobacillus casei* VB178, VB179, VB180, and VB217 against *Enterococcus faecalis* according to embodiments of the present invention. Detailed Implementation

[0023] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0024] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0025] In this document, the terms “containing,” “comprising,” or “including” are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0026] In this document, “prevention” and “avoidance” are used interchangeably. These terms refer to methods for obtaining beneficial or desired results, including but not limited to preventive benefits. To obtain a “preventive benefit,” *Lactobacillus casei* or a product containing it may be given to a subject at risk of developing a specific disease, or to a subject who reports one or more physiological symptoms of a disease, even if a diagnosis of the disease may not have been made.

[0027] In this document, the terms “treatment” and “relief” refer to the attainment of a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of the disease or its symptoms, and / or therapeutic in terms of partial or complete cure of the disease and / or adverse effects caused by the disease. As used herein, “treatment” covers diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of a condition in individuals susceptible to the disease but not yet diagnosed with it; (b) suppression of the disease, such as inhibiting disease progression; or (c) relief of the disease, such as reducing symptoms associated with the disease. As used herein, “treatment” encompasses any administration of a drug or compound to an individual to treat, cure, relieve, improve, reduce, or suppress the individual’s disease, including but not limited to administration of a drug containing a compound described herein to an individual in need.

[0028] This invention proposes Lactobacillus casei VB179, fermentation broth, bacterial suspension or bacterial residue, microbial agent, single-dose preparation, product, and their uses and cultivation or production apparatus, which will be described in detail below.

[0029] strains

[0030] In one aspect of the invention, a *Lactaseibacillus casei* VB179 is provided. According to an embodiment of the invention, the *Lactaseibacillus casei* VB179 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 28623, deposited on October 12, 2023, at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The *Lactaseibacillus casei* VB179 of the present invention exhibits broad antibacterial activity, significantly inhibiting not only the growth of *Malassezia*, but also various other pathogenic bacteria, including *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*. This provides potential therapeutic options and possibilities for further treatment of diseases associated with these pathogens.

[0031] In this article, "Lactobacillus casei VB179" and "Lactobacillus casei VB179" are synonymous.

[0032] Fermentation broth, bacterial suspension or bacterial residue

[0033] In another aspect, the present invention provides a fermentation broth, bacterial suspension, or bacterial residue. According to an embodiment of the present invention, the fermentation broth, bacterial suspension, or bacterial residue includes the previously mentioned *Lactobacillus casei* VB179. Therefore, the fermentation broth, bacterial suspension, or bacterial residue of the present invention can also significantly inhibit the growth of *Malassezia*, and thus can be used to treat skin diseases such as tinea versicolor, Malassezia folliculitis, seborrheic dermatitis, and characteristic dermatitis caused by the overgrowth of *Malassezia*.

[0034] It should be noted that the "fermentation broth" mentioned in this invention refers to the solution obtained after culturing *Lactobacillus casei* VB179 for a period of time. The "cultural residue" mentioned in this invention refers to the bacterial precipitate obtained by centrifuging the fermentation broth and removing the supernatant. The "cultural suspension" mentioned in this invention refers to the suspension obtained by adding a certain volume of physiological saline to the bacterial residue and mixing it thoroughly by pipetting.

[0035] Microbial agents

[0036] In another aspect, the present invention provides a microbial inoculant. According to embodiments of the present invention, the microbial inoculant comprises *Lactobacillus casei* VB179 as described above, or the aforementioned fermentation broth, bacterial suspension, or bacterial residue. Therefore, the microbial inoculant of the present invention not only significantly inhibits the growth of *Malassezia*, but also inhibits the growth of various other pathogenic bacteria, including *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, providing potential treatment options and possibilities for further treatment of diseases associated with these pathogenic bacteria.

[0037] It should be noted that the microbial inoculant of the present invention may be a liquid microbial inoculant, including but not limited to fermentation broth; or it may be a solid microbial inoculant, including but not limited to freeze-dried powder.

[0038] According to embodiments of the present invention, the microbial inoculant may further include at least one of the following additional technical features:

[0039] According to an embodiment of the present invention, the Lactobacillus casei VB179 exists in the form of live cells and / or non-live cells.

[0040] In this article, "living cell" refers to Lactobacillus casei VB179 with the ability to metabolize, reproduce, or replicate.

[0041] For example, the living cells may be immobilized cells. In this document, "immobilized cells" refers to Lactobacillus casei VB179 immobilized on a carrier, capable of carrying out life activities such as growth, development, reproduction, heredity, and metabolism within a certain spatial range.

[0042] In this document, "non-living cells" refers to cells that do not have the ability to metabolize, reproduce, and replicate, including but not limited to dried bacterial cells. For example, the microbial agent is a lyophilized powder.

[0043] For example, the Lactobacillus casei VB179 exists as live cells, dry cells, immobilized cells, or in any other form.

[0044] For example, the dried bacterial cells are obtained by freeze-drying the Lactobacillus casei VB179.

[0045] According to embodiments of the present invention, the microbial agent further includes pharmaceutically acceptable excipients or carriers.

[0046] In this document, "pharmaceutical acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with other components of the formulation and / or the mammals to which it is treated. Preferably, "pharmaceutical acceptable" as used herein means approved by a federal regulatory agency or national government, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals, particularly in humans.

[0047] In this document, the term "pharmaceutically acceptable carrier" includes any solvent, drug stabilizer, or combination thereof known to those skilled in the art. It covers the use of any conventional carrier in therapeutic or pharmaceutical compositions, except in cases where any conventional carrier is incompatible with the active ingredient.

[0048] In this document, the term "pharmaceuticalally acceptable excipient" may include any solvent suitable for the specific target dosage form. The use of any conventional excipients that are incompatible with the *Lactobacillus casei* VB179 disclosed herein, for example, for any adverse biological effects produced or for interactions that occur in a harmful manner with any other component of the pharmaceutically acceptable composition, is also within the scope of this invention.

[0049] Single-dose formulation

[0050] In another aspect, the present invention provides a single-dose formulation. According to embodiments of the invention, the single-dose formulation comprises not less than 10 6 CFU uses Lactobacillus casei VB179, as the active ingredient, as mentioned above. For example, it can be 10... 6 ~10 12 CFU, 106 ~10 10 CFU, 10 6 ~10 9 CFU, more specifically, can be, but is not limited to, 10. 6 CFU, 10 7 CFU, 10 8 CFU, 10 9 CFU, 10 10 CFU, 10 11 CFU, 10 12 CFU. Therefore, the single-dose formulation of the present invention not only significantly inhibits the growth of Malassezia, but also inhibits the growth of a variety of other pathogenic bacteria, including Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens, providing potential treatment options and possibilities for further treatment of diseases associated with these pathogenic bacteria.

[0051] It should be noted that the "single-dose formulation" described in this invention refers to a formulation containing a complete and accurate dose of drug in a single administration, suitable for a single use. Simply put, it is a formulation containing a single dose of drug that can be given directly to the patient without further division or measurement.

[0052] product

[0053] In another aspect, the present invention provides a product. According to embodiments of the invention, the product comprises at least one of the following: *Lactobacillus casei* VB179, the aforementioned fermentation broth, bacterial suspension or residue, the aforementioned microbial agent, and the aforementioned single-dose formulation. Thus, the product of the present invention can be used to inhibit the growth of *Malassezia*, *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, and *Clostridium perfringens*, thereby preventing and / or alleviating and / or treating diseases associated with these pathogens.

[0054] According to embodiments of the present invention, the product may further include at least one of the following additional technical features:

[0055] According to an embodiment of the present invention, the product is selected from daily chemical products or pharmaceuticals.

[0056] According to embodiments of the present invention, when the product is selected from pharmaceuticals, it may further include excipients and / or carriers.

[0057] According to embodiments of the present invention, the excipient includes at least one selected from adhesives, disintegrants, lubricants, flow aids, stabilizers, fillers, diluents, and sustained-release agents.

[0058] According to an embodiment of the present invention, the carrier comprises at least one selected from sugars, cellulose and its derivatives, calcium phosphates, alkaline earth metal stearates, vegetable oils, nonionic surfactants, cationic surfactants, anionic surfactants, fatty alcohols, and hydrolyzed solids of grains.

[0059] According to embodiments of the present invention, the dosage forms of the drug include, but are not limited to, oral preparations, injections, external solutions, lotions, granules, ointments, patches, and smears.

[0060] According to embodiments of the present invention, the daily chemical products include, but are not limited to, shampoos, shower gels, conditioners, soaps, face creams, skin creams, lotions, body lotions, and toners.

[0061] use

[0062] In another aspect of the invention, the invention proposes the use of the aforementioned *Lactobacillus casei* VB179, the aforementioned fermentation broth, bacterial suspension or residue, the aforementioned microbial agent, or the aforementioned single-dose formulation in the preparation of a medicament for the prevention and / or relief and / or treatment of related diseases or symptoms caused by pathogenic bacterial infections.

[0063] According to embodiments of the present invention, the use may further include at least one of the following additional technical features:

[0064] According to an embodiment of the present invention, the pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

[0065] According to an embodiment of the present invention, the relevant disease is selected from skin diseases.

[0066] According to embodiments of the present invention, the skin diseases include, but are not limited to, tinea versicolor, pityriasis folliculitis, seborrheic dermatitis, atopic dermatitis, and psoriasis.

[0067] According to embodiments of the present invention, the symptoms include, but are not limited to, dandruff.

[0068] In another aspect of the invention, the invention provides the use of the aforementioned Lactobacillus casei VB179, the aforementioned fermentation broth, bacterial suspension or bacterial residue, the aforementioned microbial agent or the aforementioned single-dose formulation in inhibiting the growth of pathogenic bacteria.

[0069] According to an embodiment of the present invention, the pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

[0070] In another aspect of the invention, the invention proposes the use of the aforementioned Lactobacillus casei VB179, the aforementioned fermentation broth, bacterial suspension or bacterial residue, the aforementioned microbial agent or the aforementioned single-dose formulation for the prevention and / or relief and / or treatment of related diseases or symptoms caused by pathogenic bacterial infections.

[0071] According to an embodiment of the present invention, the pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

[0072] According to an embodiment of the present invention, the relevant disease is selected from skin diseases.

[0073] According to embodiments of the present invention, the skin diseases include, but are not limited to, tinea versicolor, pityriasis folliculitis, seborrheic dermatitis, atopic dermatitis, and psoriasis.

[0074] According to embodiments of the present invention, the symptoms include, but are not limited to, dandruff.

[0075] method

[0076] In another aspect, the present invention provides a method for inhibiting the growth of pathogenic bacteria in vitro. According to an embodiment of the present invention, the method includes: co-culturing at least one of the aforementioned *Lactobacillus casei* VB179, the aforementioned fermentation broth, bacterial suspension or bacterial residue, the aforementioned microbial agent, and the aforementioned single-dose formulation with a sample containing pathogenic bacteria. As previously stated, the *Lactobacillus casei* VB179 of the present invention has broad-spectrum antibacterial activity and can effectively inhibit a variety of pathogenic bacteria. Therefore, the method described in the present invention can efficiently prevent the growth of pathogenic bacteria, thereby effectively controlling the infections and diseases they cause, providing an efficient and reliable approach for the treatment and prevention of diseases.

[0077] According to an embodiment of the present invention, the pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

[0078] In another aspect, the present invention provides a method for preventing and / or alleviating and / or treating diseases or symptoms caused by pathogenic bacterial infections. According to embodiments of the present invention, the method comprises administering to a patient at a pharmaceutically acceptable dose of at least one of the following: *Lactobacillus casei* VB179 as described in the first aspect, fermentation broth, bacterial suspension or bacterial residue as described in the second aspect, microbial agent as described in the third aspect, or single-dose formulation as described in the fourth aspect. As previously stated, *Lactobacillus casei* VB179 of the present invention has broad-spectrum antibacterial activity and can effectively inhibit a variety of pathogenic bacteria. Therefore, the method described in the present invention can efficiently prevent the growth of pathogenic bacteria, thereby effectively controlling the infections and diseases they cause, providing an efficient and reliable approach for the treatment and prevention of diseases.

[0079] According to an embodiment of the present invention, the pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

[0080] According to an embodiment of the present invention, the relevant disease is selected from skin diseases.

[0081] According to embodiments of the present invention, the skin diseases include, but are not limited to, tinea versicolor, pityriasis folliculitis, seborrheic dermatitis, atopic dermatitis, and psoriasis.

[0082] According to embodiments of the present invention, the symptoms include, but are not limited to, dandruff.

[0083] Device

[0084] In another aspect, the present invention provides a culture or production apparatus for the aforementioned *Lactobacillus casei* VB179, the aforementioned fermentation broth, bacterial suspension or residue, the aforementioned microbial agent, or the aforementioned single-dose formulation. The culture apparatus of the present invention can be used for the mass production of *Lactobacillus casei* VB179 or its fermentation products, facilitating the development of production apparatus for microbial agents or single-dose formulations, enabling the mass production of commercial products, and possessing high economic value.

[0085] The embodiments of the present invention are described in detail below. These embodiments are exemplary and are only used to explain the present invention, and should not be construed as limiting the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. Reagents or instruments used, unless otherwise specified, are all commercially available conventional products.

[0086] Unless otherwise specified, all percentages (%) used in the culture media in the following examples are mass percentages.

[0087] Example 1: Identification of Lactobacillus casei strains

[0088] (1) Source of strains

[0089] Lactobacillus casei was isolated from yogurt. Sufficient yogurt was placed in a sterile container with sterile PBS buffer, shaken thoroughly, diluted, and spread onto MRS solid agar plates. After 48 hours of incubation, single colonies of varying sizes and shapes were picked and streaked onto fresh plates. Then, single colonies were picked onto slant agar plates. After successful slant incubation, colony morphology was examined under a microscope. Identification was performed after confirming the absence of contaminating bacteria.

[0090] (2) Strain identification

[0091] The experiments were conducted according to the relevant content in the book *Molecular Cloning: A Laboratory Manual*. After culturing the purified single bacterial strain in MRS liquid medium for one day, 1 mL of the bacterial culture was centrifuged at 10,000 rpm for 1 min, the supernatant was discarded, and the bacterial cells were collected. DNA was extracted according to the procedures of the MAGEN Bacterial Genomic DNA Extraction Kit (purchased from Shanghai Sangon Biotech Co., Ltd.). Using the extracted DNA as a template, 16S rRNA amplification was performed by PCR (polymerase chain reaction) using universal primers 337F (5'-GACTCCTACGGGAGGCWGCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'). The PCR reaction volume was 50 μL, and the reaction system and program were prepared according to the instructions of Phusion high-fidelity DNA polymerase (purchased from Thermo Scientific), with an annealing temperature of 55℃. The PCR products were sent to Shanghai Sangon Biotech Co., Ltd. for sequencing.

[0092] After the 16S rRNA sequence of the strain was proofread, it was compared with the homologous sequence BLAST of the sequences of related species and genera in the GenBank database to determine the taxonomic position of the strain.

[0093] Comparison revealed that it shared 99.93% homology with *Lactobacillus casei* strain NWAFU1563 and 99.93% homology with *Lacticaseibacillus casei* strain D2. Therefore, this strain was identified as *Lacticaseibacillus casei*. The internal code for this strain is VB179. It was deposited on October 12, 2023, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 28263.

[0094] The 16S rRNA sequencing results of Lactobacillus casei VB179 are shown in SEQ ID NO:1:

[0095] Example 2: Physiological and biochemical experiments of Lactobacillus casei strain

[0096] 1. Preparation of Lactobacillus casei

[0097] (1) Culture medium: 10g tryptone, 10g beef extract, 10g yeast extract, 2.0g disodium hydrogen citrate, 20g glucose, 1mL Tween 80, 5.0g sodium acetate, 2.0g dipotassium hydrogen phosphate, 0.58g magnesium sulfate, and 0.25g manganese sulfate were added to 1L of distilled water and sterilized in an autoclave at 115℃ for 20min to obtain fermentation liquid culture medium; 10g tryptone, 10g beef extract, 10g yeast extract, 2.0g disodium hydrogen citrate, 20g glucose, 1mL Tween 80, 5.0g sodium acetate, 2.0g dipotassium hydrogen phosphate, 0.58g magnesium sulfate, 0.25g manganese sulfate, and 20g agar were added to 1L of distilled water and sterilized in an autoclave at 115℃ for 20min to obtain MRS solid culture medium.

[0098] (2) Activation: Take out the Lactobacillus casei frozen in a -80℃ freezer, lightly dip it onto MRS solid medium and streak it. After 24 hours in a 37℃ incubator, obtain a single colony.

[0099] 2. Physiological and biochemical experiments

[0100] (1) Physiological and biochemical reagents: HBI lactic acid bacteria biochemical identification strips were purchased from Qingdao Haibo Biotechnology Co., Ltd. These strips are used for the biochemical identification of lactic acid bacteria. Each strip contains: aescin, cellobiose, maltose, mannitol, salicin, sorbitol, sucrose, raffinose, inulin, lactose, and hippuric acid, totaling 11 biochemical reactions.

[0101] (2) Experimental preparation: Take one biochemical identification strip from the packaging box, open the cap, and make a hole with a punch or simply tear off the membrane. If it is contaminated or discolored, it cannot be used; take a new strip. Note: Wipe the punch with 75% alcohol before use.

[0102] (3) Inoculation:

[0103] Select the Lactobacillus casei from step 1, prepare a bacterial suspension, and inoculate it into the following culture media: aescin, cellobiose, maltose, mannitol, maltose, salicin, sorbitol, sucrose, raffinose, inulin, lactose, and 1% sodium hippurate.

[0104] Bacterial suspension method: Take a test tube containing 2 mL of sterile physiological saline, pick a single colony from the purified culture plate with an inoculation needle, grind it carefully in sterile physiological saline to prepare a homogeneous bacterial suspension, and add 100 μl of bacterial suspension to each well.

[0105] (4) Culture: After inoculation, mark the inoculation, cover it, place it in the base tray, and incubate at 36±1℃.

[0106] (5) Observation: After 24 hours, the culture is completed and placed on the record card for observation.

[0107] The experimental results are shown in Table 1 below. It can be seen that the strain of Lactobacillus casei VB179 can utilize cellobiose, maltose, mannitol, salicin, sorbitol, sucrose, inulin, lactose, and 1% sodium hippurate; but cannot utilize aescin and raffinose.

[0108] Table 1: Results of the VB179 Physiological and Biochemical Reagent Kit

[0109] Example 3: Inhibitory effect of Lactobacillus casei on Malassezia

[0110] 1. Lactobacillus casei

[0111] (1) Culture medium: 10g tryptone, 10g beef extract, 10g yeast extract, 2.0g disodium hydrogen citrate, 20g glucose, 1mL Tween 80, 5.0g sodium acetate, 2.0g dipotassium hydrogen phosphate, 0.58g magnesium sulfate, and 0.25g manganese sulfate were added to 1L of distilled water and sterilized in an autoclave at 115℃ for 20min to obtain fermentation liquid culture medium; 10g tryptone, 10g beef extract, 10g yeast extract, 2.0g disodium hydrogen citrate, 20g glucose, 1mL Tween 80, 5.0g sodium acetate, 2.0g dipotassium hydrogen phosphate, 0.58g magnesium sulfate, 0.25g manganese sulfate, and 20g agar were added to 1L of distilled water and sterilized in an autoclave at 115℃ for 20min to obtain MRS solid culture medium.

[0112] (2) Activation: Take out the Lactobacillus casei frozen at -80℃, lightly dip it onto MRS solid medium and streak it, then place it in an incubator at 37℃ for 24 hours. Scratch a single colony from the MRS solid medium and place it in fermentation liquid medium. After culturing at 37℃ and 180 rpm for 24 hours, the activation is complete.

[0113] 2. Malassezia

[0114] (1) Malassezia was purchased from Shanghai Preservation Biotechnology Center.

[0115] (2) Activation: Commercially available YM liquid medium was used. For YM solid medium, an additional 2% agar was added to the liquid medium. A small amount of Malassezia (frozen at -80℃) was streaked onto the YM solid medium and incubated at 30℃ for 48 hours to complete activation. The YM medium consisted of: 0.3% yeast extract, 0.3% malt extract, and 0.5% peptone.

[0116] 3. In vitro inhibition experiment:

[0117] (1) Treatment of Lactobacillus casei: Take activated Lactobacillus casei and inoculate it into the fermentation liquid culture medium at an inoculation ratio of 1%. After culturing at 30℃ and 180rpm for 48h, take it out. Centrifuge the obtained fermentation broth at 4℃ and 4000rpm for 10min in a refrigerated centrifuge. The supernatant after centrifugation is the fermentation supernatant, and the bacterial cells are the bacterial residue. Add the bacterial residue to the same volume of physiological saline, blow to break the cells and mix well to obtain the cell suspension.

[0118] The original fermentation broth was diluted to an appropriate factor to obtain a diluted solution. 100 μL of this diluted solution was then spread onto MRS solid medium for counting.

[0119] (2) Malassezia treatment: Take a small amount of activated Malassezia single colony, place it in an appropriate amount of sterile physiological saline, and shake to mix. Measure the concentration of Malassezia using a hemocytometer, and then adjust the concentration to approximately 1.0 × 10⁻⁶. 6 CFU / mL.

[0120] In vitro antibacterial experiment: The double-layer plate culture method was used. The optimized and evaluated method is as follows: The double-layer plate culture method was used, and the method is as follows:

[0121] ① Preparation of the lower layer: Pour in MRS solid medium. Inoculate the activated *Lactobacillus casei* fermentation broth, mycelial residue, and cytostomated bacterial suspension onto the lower layer medium. Inoculation amount: 2 μl / inoculation (10⁻⁶). 8 (CFU count); after anaerobic incubation at 37 degrees Celsius for 1 day, pour in the upper layer of pathogenic bacteria.

[0122] ② Preparation of the upper pathogenic bacteria layer: After the upper culture medium (YM solid medium) has been sterilized at 121℃ for 30 minutes, it is cooled to 40-50℃. Malassezia is added as needed (the final concentration of pathogenic bacteria in the culture medium is approximately 10). 6 Mix well (CFU / ml), then add 7ml / plate of the culture medium containing Malassezia to the above YM solid medium (this medium is a blank plate). After solidification, add 2μl / point of positive control (0.1mg / L ketoconazole) and 2μl / point of water. Incubate at 37℃ under aerobic conditions for 1 day and observe whether there is an inhibition zone around the probiotics to determine whether it has antibacterial ability.

[0123] The results of the external plate inhibition experiment are shown in Figure 1. It is evident that the inhibition zone of *Lactobacillus casei* fermentation broth against *Malaiseibacterium* is significantly larger than that of ketoconazole at a concentration of 0.1 mg / L. However, the inhibition zones of *Lactobacillus casei* residue and suspension against *Malaiseibacterium* are not significantly different from those of 0.1 mg / L ketoconazole. This indicates that *Lactobacillus casei* residue, fermentation broth, and suspension all exhibit good inhibitory effects on *Malaiseibacterium*, with the fermentation broth showing a greater inhibitory effect than ketoconazole at that concentration (0.1 mg / L).

[0124] Example 4: Inhibitory effect of different Lactobacillus casei suspensions on Malassezia

[0125] The inventors also studied the inhibitory effects of VB178, VB180, and VB217 (16S rRNA sequencing results of VB178, VB180, and VB217 are shown in SEQ ID NO:1) on Malassezia, strains belonging to the same genus but different from VB179. Following the method described in Example 3, the inventors simultaneously conducted in vitro antibacterial experiments on VB178, VB179, VB180, and VB217. The inoculum size for different strains was 2 μl / point (10... 6 The bacterial count (CFU) was measured, and the bacteria were cultured at 37°C under aerobic conditions for 1 day. The size of the inhibition zone around different types of Lactobacillus casei was observed to determine whether they had antibacterial ability.

[0126] The experimental results are shown in Figure 2. It can be clearly seen that although VB178, VB180 and VB217 also have a certain inhibitory effect on Malassezia, their inhibitory effect is significantly weaker than that of VB179. Therefore, the antibacterial ability of Lactobacillus casei VB179 is significantly higher than that of other strains of Lactobacillus casei.

[0127] Example 5: Inhibitory effects of different types of Lactobacillus casei on other pathogenic bacteria

[0128] To investigate whether different *Lactobacillus casei* strains, besides inhibiting *Malassezia*, also inhibit other pathogenic bacteria, the inventors used a double-layer plate culture method to study the inhibitory effects of VB178, VB179, VB180, and VB217 on nine pathogenic bacteria, including *Candida albicans*, *Clostridium perfringens*, *Gardnerella vaginalis*, *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Shigella*, and *Listeria*. The specific methods are as follows:

[0129] ① Preparation of *Lactobacillus casei* from the lower layer: Different *Lactobacillus casei* strains VB178, VB179, VB180, and VB217 were activated as in Example 3-1 and inoculated into fermentation broth at a 1% inoculation ratio. The cultures were incubated at 30°C and 180 rpm for 48 hours. The resulting fermentation broth was centrifuged at 4°C and 4000 rpm for 10 minutes using a refrigerated centrifuge. The supernatant obtained after centrifugation was the fermentation supernatant, and the bacterial cells were the bacterial residue. The bacterial residue was mixed with an equal volume of physiological saline by pipetting to obtain the lysed bacterial suspension.

[0130] The bacterial suspensions of the above different strains were inoculated at an inoculation rate of 2 μl / point (10 6 (CFU count) is inoculated onto a prepared MRS solid petri dish, anaerobic at 37°C, and cultured for 1 day before the upper layer of pathogenic bacteria is poured in.

[0131] ② Preparation of the upper pathogenic bacteria layer: After sterilizing the upper culture medium at 121℃ for 30 minutes (the upper culture medium is the culture medium required for different pathogens, as shown in Table 2), cool it to 40-50℃, and add the different pathogens as needed (the final concentration of pathogens in the culture medium is approximately 10). 6 Mix well (CFU / ml), add 7ml / plate of the culture medium containing pathogenic bacteria to the single-cell plates of Lactobacillus casei VB178, VB179, VB180, and VB217 that have been cultured for 1 day. After solidification, culture according to the growth conditions of different pathogenic bacteria (see Table 2 for culture methods). After the upper plate has grown well, observe whether there is an inhibition zone around the probiotics to determine whether it has antibacterial ability.

[0132] Table 2: Culture conditions for different pathogenic bacteria

[0133] The inhibition zone sizes of different *Lactobacillus casei* strains are shown in Table 3. It can be seen that *Lactobacillus casei* VB179 exhibits greater inhibitory activity against common bacteria such as *Salmonella*, *Enterococcus faecalis* (inhibition zones of different *Lactobacillus casei* strains against *Enterococcus faecalis* are shown in Figure 3), *Listeria*, *Shigella*, and *Gardnerella* than other *Lactobacillus casei* strains. Furthermore, *Lactobacillus casei* VB179's inhibitory activity against *Escherichia coli*, *Clostridium perfringens*, *Staphylococcus aureus*, and *Candida albicans* is also comparable to other *Lactobacillus casei* strains. Therefore, the *Lactobacillus casei* VB179 of this invention not only has a strong inhibitory effect on *Malassezia*, but can also simultaneously inhibit the growth of *Escherichia coli*, *Staphylococcus aureus*, *Salmonella*, *Enterococcus faecalis*, *Listeria*, *Shigella*, *Clostridium perfringens*, *Candida albicans*, and *Gardnerella*.

[0134] Table 3: Size of inhibition zones of different Lactobacillus casei against different pathogenic bacteria

[0135] Example 6: Animal Experiment

[0136] 1. Laboratory animals: 20 white, clean-grade male guinea pigs, weighing 300-320g, were raised in our animal facility. All guinea pigs were individually housed in stainless steel cages at a room temperature of 25±2℃.

[0137] 2. Experimental strain: Malassezia and culture medium: see Example 3.

[0138] 3. Method:

[0139] (1) Preparation of bacterial suspension

[0140] To ensure viability, *Malassezia spheroidosa* was passaged twice on YM solid medium. After the second incubation at 31°C for 3-4 days, colonies were picked and placed in 0.9% physiological saline. The colonies were repeatedly pipetted using a sterile pipette to prepare a *Malassezia spheroidosa* suspension. The suspension was then counted on a hemocytometer, and the final density was adjusted to 1 × 10⁻⁶. 6 CFU / mL.

[0141] (2) Preparation of bacteria and drugs

[0142] The bacterial solution was prepared into a 2% petroleum jelly cream, and the negative control was simple petroleum jelly cream.

[0143] (3) Establishment of a guinea pig dermal Malassezia infection animal model: After shaving the back of guinea pigs, hair removal was performed (using beeswax: rosin = 1:1) to create a 4.5cm × 7.5cm hairless area. Direct microscopic examination and culture of fungi were performed on the hairless area of ​​the guinea pig's back for 7 days, and the results were all negative. The guinea pig's back was gently and evenly rubbed with sandpaper, and 200μL of the above-mentioned Malassezia bacterial suspension was evenly applied to the hairless area of ​​the back once a day. For 7 consecutive days, the Malassezia bacterial suspension was freshly prepared daily, and colonies were collected from the same culture dish. One day after the last inoculation, skin samples from both the inoculated and uninoculated areas were taken, fixed in 10% formaldehyde solution, embedded in paraffin, sectioned, and observed after PAS staining.

[0144] (4) In the Malassezia infection experiment in guinea pigs, a self-control design was adopted. The backs of the guinea pigs were evenly divided into two rows, each row containing three 1.5 cm diameter circles. Each circle was spaced approximately 1.5 cm apart to prevent interference from different treatment drugs. Each circle represented an independent experimental group, for a total of two experimental groups:

[0145] Treatment group: Topical application of Lactobacillus casei VB179 to the skin lesions;

[0146] Negative control group: Simple petroleum jelly cream was applied topically to the skin lesions.

[0147] 4. Evaluation of drug efficacy

[0148] One day after the end of drug administration, guinea pigs in each group were euthanized for microscopic examination of fungi. Before the examination, the back area of ​​the experimental guinea pigs was thoroughly disinfected with 75% alcohol swabs. Then, circular samples for each experiment were cut and collected. Next, the samples were finely chopped and placed in a glass mixer containing 2 ml of 0.9% sterile physiological saline, with 0.5% sorbitol added to form a homogeneous suspension. 300 μl of this suspension was transferred to a round petri dish containing solid culture medium, immediately shaken to ensure even distribution on the surface of the medium, and incubated at 32°C for 3 days. The number of colonies in the petri dish was observed; if the number of colonies in the petri dish was >1, it was considered a positive culture. The fungal conversion rate for each group was calculated.

[0149] Mycological cure: Direct microscopic examination and culture of fungi were both negative.

[0150] Fungal clearance rate: the number of mycologically cured guinea pigs in each group divided by the total number of guinea pigs in each group.

[0151] Table 4 shows the fungal clearance rates in each group after drug treatment. It is evident that the fungal clearance rate in the treatment group (with topical application of Lactobacillus casei VB179 to the skin lesions) was significantly higher than that in the negative control group. This indicates that Lactobacillus casei VB179 has a significant inhibitory effect on Malassezia in animal experiments.

[0152] Table 4: Fungal clearance rate in each group of guinea pigs after drug treatment (n=10)

[0153] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0154] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A type of Lactaseibacillus casei VB179, characterized in that, The Lactobacillus casei VB179 was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 28623 on October 12, 2023.

2. A fermentation broth, bacterial suspension, or bacterial residue, characterized in that, Includes Lactobacillus casei VB179 as described in claim 1.

3. A microbial inoculant, characterized in that, Includes Lactobacillus casei VB179 as described in claim 1 or fermentation broth, bacterial suspension or bacterial residue as described in claim 2.

4. The microbial agent according to claim 3, characterized in that, This further includes pharmaceutically acceptable excipients or carriers.

5. A single-dose formulation, characterized in that, Including no less than 10 6 CFU uses Lactobacillus casei VB179 as the active ingredient as described in claim 1.

6. The use of the *Lactobacillus casei* VB179 of claim 1, the fermentation broth, bacterial suspension or bacterial residue of claim 2, the microbial agent of any one of claims 3 to 4, or the single-dose formulation of claim 5 in the preparation of a medicament, wherein the medicament is used to prevent and / or alleviate and / or treat related diseases or symptoms caused by pathogenic bacterial infection.

7. The use according to claim 6, characterized in that, The pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

8. The use according to claim 6, characterized in that, The relevant disease or symptom is selected from at least one of tinea versicolor, pityrosporum folliculitis, seborrheic dermatitis, dandruff, atopic dermatitis, and psoriasis.

9. A product characterized in that, It includes at least one of the following: Lactobacillus casei VB179 as described in claim 1, fermentation broth, bacterial suspension or bacterial residue as described in claim 2, microbial agent as described in any one of claims 3 to 4, and single-dose formulation as described in claim 5.

10. The product according to claim 9, characterized in that, The products are selected from daily chemical products or pharmaceuticals.

11. A method for inhibiting the growth of pathogenic bacteria in vitro, characterized in that, include: At least one of the following is co-cultured with a sample containing pathogenic bacteria: Lactobacillus casei VB179 of claim 1, fermentation broth, bacterial suspension or bacterial residue of claim 2, microbial agent of any one of claims 3 to 4, and single-dose formulation of claim 5.

12. The method according to claim 11, characterized in that, The pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

13. A culture or production apparatus for Lactobacillus casei VB179 as described in claim 1, the fermentation broth, bacterial suspension or bacterial residue as described in claim 2, the microbial agent as described in any one of claims 3 to 4, or the single-dose formulation as described in claim 5.

14. Use of the Lactobacillus casei VB179 of claim 1, the fermentation broth, bacterial suspension or bacterial residue of claim 2, the microbial agent of any one of claims 3 to 4, or the single-dose formulation of claim 5 in inhibiting the growth of pathogenic bacteria.

15. The *Lactobacillus casei* VB179 of claim 1, the fermentation broth, bacterial suspension or bacterial residue of claim 2, the microbial agent of any one of claims 3-4, or the single-dose formulation of claim 5, for the prevention and / or relief and / or treatment of related diseases or symptoms caused by pathogenic bacterial infections.

16. The use according to claim 14 or 15, characterized in that, The pathogenic bacteria are selected from at least one of Malassezia, Escherichia coli, Staphylococcus aureus, Salmonella, Enterococcus faecalis, Listeria monocytogenes, Shigella, and Clostridium perfringens.

17. The use according to claim 15, characterized in that, The relevant disease or symptom is selected from at least one of tinea versicolor, pityrosporum folliculitis, seborrheic dermatitis, dandruff, atopic dermatitis, and psoriasis.

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