Composition for inhibiting harmful bacteria in the skin and oral cavity

Abstract

To provide lactic acid bacteria that are highly effective in inhibiting harmful bacteria in the skin and oral cavity.SOLUTION: A composition for inhibiting harmful bacteria in the skin and oral cavity is used for application to the skin and oral cavity, the composition containing heat-treated bacterial cells of Lactiplantibacillus plantarum HJ1 (lactic acid bacteria HJ1 strain) as an active ingredient. Preferably, the composition has the ability to reduce odors in the skin or oral cavity to which it is applied.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to compositions for use on the skin and in the oral cavity for inhibiting harmful bacteria. [Background technology]

[0002] In recent years, it has been thought that maintaining a balance in the intestinal flora is important for maintaining good health, and the same is true for the skin, and it is believed that by maintaining a balance in the skin flora, good skin health can be maintained.

[0003] Regarding such a technique, for example, Patent Document 1 discloses that 1-ketose is used to promote the growth of Staphylococcus epidermidis, which is known as a beneficial bacterium, while not promoting the growth of Staphylococcus aureus, which is known as a harmful bacterium.

[0004] On the other hand, Patent Document 2 discloses a substance that exhibits a growth inhibitory effect against Streptococcus mutans, which is known to cause dental caries. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2022-184273 A [Patent Document 2] Patent Publication No. 2022-27681 Summary of the Invention [Problem to be solved by the invention]

[0006] To date, little research has been done on the use of lactic acid bacteria to suppress harmful bacteria on the skin or in the oral cavity.

[0007] An object of the present invention is to provide a lactic acid bacterium that is highly effective in suppressing harmful bacteria on the skin and in the oral cavity. [Means for solving the problem]

[0008] As a result of various studies conducted to achieve the above-mentioned object, the inventors discovered that lactic acid bacteria HJ1 strain (zygosaccharomyces cerevisiae) has the effect of suppressing harmful bacteria on the skin and oral cavity, and thus completed the present invention.

[0009] That is, the present invention provides a composition for inhibiting harmful bacteria on the skin and in the oral cavity, which contains heat-treated cells of Lactiplantibacillus plantarum HJ1 (lactic acid bacteria HJ1 strain (zygosaccharomyces cerevisiae)) as an active ingredient and is applied to the skin or oral cavity.

[0010] In the composition of the present invention, the harmful bacteria is preferably one or more species selected from the group consisting of Staphylococcus aureus, Staphylococcus pseudintermedius, Streptococcus mutans, Porphyromonas macacae, Candida albicans, Malassezia furfur, and Helicobacter pylori.

[0011] Furthermore, the composition of the present invention preferably has the effect of reducing odor on the skin or oral cavity to which it is applied. Effect of the Invention

[0012] According to the present invention, lactic acid bacteria HJ1 strain (zygote) can be used to provide lactic acid bacteria that are highly effective in suppressing harmful bacteria on the skin and in the oral cavity. [Brief description of the drawings]

[0013] [Figure 1]1 is a chart showing the results of investigating the effect of lactic acid bacteria HJ1 strain on the proliferation of Staphylococcus aureus in Test Example 1. [Diagram 2] 1 is a chart showing the results of investigating the effect of lactic acid bacteria HJ1 strain on the proliferation of Staphylococcus pseudintermedius, a causative bacterium of pyoderma, in Test Example 2. [Diagram 3] 1 is a chart showing the results of investigating the effect of lactic acid bacteria HJ1 strain on the proliferation of Streptococcus mutans, a causative bacterium of dental caries, in Test Example 3. [Figure 4] 1 is a chart showing the results of investigating the effect of lactic acid bacteria HJ1 strain on the proliferation of Staphylococcus epidermidis, a normal skin flora, in Test Example 4. [Diagram 5] 1 is a graph showing the results of investigating the odor reducing effect when lactic acid bacteria HJ1 strain was applied to the human oral cavity in Test Example 5. [Figure 6] 1 is a graph showing the results of investigating the odor reducing effect when lactic acid bacteria HJ1 strain is applied to human skin in Test Example 6. [Figure 7] 1 is a graph showing the results of investigating the odor reducing effect when lactic acid bacteria HJ1 strain was applied to the oral cavity of a dog in Test Example 7. [Figure 8] 1 is a graph showing the results of investigating the odor reducing effect when lactic acid bacteria HJ1 strain was applied to the oral cavity of a cat in Test Example 8. [Figure 9] 1 is a graph showing the results of an investigation into the immune activation ability of the lactic acid bacteria HJ1 strain by a test for measuring the amount of IL-12 produced using mouse spleen cells in Test Example 9. [Figure 10]1 is a chart showing the results of investigating the effect of lactic acid bacteria HJ1 strain on the proliferation of Porphyromonas macacae, a causative bacterium of periodontal disease, in Test Example 10. [Figure 11] 1 is a chart showing the results of investigating the effect of lactic acid bacteria HJ1 strain on the proliferation of Candida albicans, a causative bacterium of candidiasis, in Test Example 11. [Figure 12] 1 is a chart showing the results of investigating the effect of Lactobacillus casei HJ1 strain on the proliferation of Malassezia furfur, a causative bacterium of tinea versicolor and seborrheic dermatitis, in Test Example 12. [Figure 13] 1 is a chart showing the results of investigating the effect of Lactobacillus strain HJ1 on the proliferation of Helicobacter pylori, a causative bacterium of gastritis and peptic ulcer, in Test Example 13. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The lactic acid bacteria used in the present invention are Lactiplantibacillus plantarum, known as a plant lactic acid bacteria derived from pickles. The scientific name is Lactiplantibacillus plantarum, and more specifically, it is Lactiplantibacillus plantarum HJ1 (Accession Number: NITE BP-03883), deposited at the National Institute of Technology and Evaluation, Patent Microorganism Depositary Center (NPMD). Hereinafter, this lactic acid bacteria strain may be referred to as "Lactic Acid Bacteria HJ1 Strain (Jinsankin, Jinsankin)".

[0015] The culture of the Lactobacillus strain HJ1 (nucleotidic acid bacteria), the maintenance of the cells, etc. can be performed by known means. For example, the medium includes a liquid medium containing a skim milk medium, yeast extract, peptone, meat extract, salts, and minerals. Commercially available media such as "MRS Bouillon MERCK" (product name, Chemicals Co.) and "Difco Lactobacilli MRS Broth" (product name, Becton Dickinson Japan Co., Ltd.) may be used. The culture can be performed, for example, by inoculating the cells into the above-mentioned medium and culturing them stationarily or culturing them with aeration and agitation under conditions of, for example, 25 to 35°C. For preservation of live cells, they can be stored in a refrigerator while suspended in the medium for a short period of time, or frozen and stored after being suspended in an antifreeze such as a glycerol solution for a long period of time.

[0016] In general, when culturing lactic acid bacteria, the pH level drops due to metabolic products of the lactic acid bacteria (such as lactic acid). To avoid this, an alkaline agent may be added to the medium to adjust the pH level (neutral culture). Examples of the alkaline agent to be added include aqueous solutions of sodium hydroxide, potassium hydroxide, calcium hydroxide, and the like, and ammonia. The pH level during culture may be in the range of 5.0 to 7.5, or may be in the range of 6.0 to 7.0, and it is preferable to adjust and maintain the pH level. The pH level may be adjusted manually, but it is simple and accurate if an automatic pH control device (pH stat) is used. However, the lactic acid bacteria used in the present invention may be any lactic acid bacteria HJ1 strain (a zymobasidium) as long as the cells are obtained, and there is no particular restriction on how to culture and prepare the lactic acid bacteria.

[0017] When preparing the lactic acid bacteria HJ1 strain, a bacterial cell concentrate can be prepared by concentrating the culture solution after culturing, or by collecting the bacteria by means of centrifugation, filtration, or the like, washing the bacteria with purified water, etc., and suspending the bacteria in purified water, etc. to a predetermined bacterial cell concentration. The content of the bacteria of the lactic acid bacteria HJ1 strain (jojoba) in 100% by mass of the bacterial cell concentrate may be in the range of 0.1 to 30% by mass, 0.5 to 20% by mass, or 1 to 10% by mass, calculated as dry bacteria. This bacterial cell concentrate may contain an excipient. This makes it easier to maintain the properties of the bacteria after freezing or lyophilization, and after reconstitution with water.

[0018] The excipient is not particularly limited, and examples thereof include dextrin; maltodextrin; xanthan gum; sugar alcohols such as lactitol, maltitol, mannitol, sorbitol, and xylitol; sugars such as dextrose, fructose, glucose, lactose, and sucrose; and organic acids such as adipic acid, citric acid, fumaric acid, glutaric acid, malic acid, succinic acid, and tartaric acid.

[0019] The form of the lactic acid bacteria HJ1 strain (joint acid bacteria) used in the present invention is preferably a heat-treated bacterial cell. In the case of heat-treated bacterial cells, the activity of live lactic acid bacteria is usually lost, so that the quality change associated with live bacteria can be suppressed. For example, the heat treatment can be carried out by culturing the lactic acid bacteria HJ1 strain (joint acid bacteria) for preparation, and then subjecting the culture solution to autoclaving as it is, or, if necessary, preparing the culture solution into the above-mentioned bacterial cell concentrate.

[0020] In another embodiment, the lactic acid bacteria HJ1 strain (jojoba) may be subjected to a pulverization / dispersion treatment. The pulverization / dispersion treatment can be carried out, for example, by pulverizing / dispersing the above-mentioned bacterial cell concentrate using a means such as stirring, a mixer, a homogenizer, a ball mill, a bead mill, a jet mill, or a generator. In this case, depending on the case or as required, the above-mentioned excipient may be added and then subjected to a pulverization / dispersion treatment to prevent reagglomeration of the obtained bacterial powder. When an excipient is contained, the content thereof may be in the range of 10 to 99% by mass, 20 to 95% by mass, or 50 to 90% by mass calculated on a dry matter basis.

[0021] In yet another embodiment, the lactic acid bacteria HJ1 strain (lactic acid bacteria) may be subjected to a process of drying and powdering, and examples of the drying and powdering process include freeze-drying, reduced pressure spray drying, spray drying using hot air, etc. By performing spray drying using hot air (spray drying), the activity of live lactic acid bacteria is usually destroyed by the heat treatment, and killed bacteria can be obtained.

[0022] As can be seen from the results of the Examples described below, the heat-treated cells of Lactobacillus sp. HJ1 strain (lactic acid bacteria) that can be prepared as described above do not show much inhibitory effect on the growth of Staphylococcus epidermidis, which is known as a beneficial bacterium of the skin, but have the effect of inhibiting the growth of harmful bacteria such as Staphylococcus aureus, Bacillus pseudointermedius which is the causative bacterium of pyoderma, Streptococcus mutans which is the causative bacterium of dental caries, Porphyromonas maccae which is the causative bacterium of periodontal disease, Candida fungi which are the causative bacteria of candidiasis, M. versicolor which is the causative bacterium of tinea versicolor and seborrheic dermatitis, and Helicobacter pylori which is the causative bacterium of gastritis and peptic ulcers.

[0023] Therefore, the present invention provides a composition for inhibiting harmful bacteria on the skin and in the oral cavity, which contains heat-treated cells of Lactobacillus casei HJ1 strain (zygosaccharomyces cerevisiae) as an active ingredient and is applied to the skin or oral cavity, and can be used, for example, as a composition that has an effect of reducing odor on the skin or oral cavity to which it is applied.

[0024] The amount of the composition of the present invention to be applied to the skin or oral cavity may be appropriately determined depending on the health condition and age of the subject, or the degree of inhibition of harmful bacteria that is required. Typically, the amount of Lactobacillus casei HJ1 strain (lactic acid bacteria) applied to the local area is 0.001 mg to 100 mg / day / cm2 in terms of dry matter. 2 and may range from 0.01 mg to 20 mg / day / cm 2 and may range from 0.1 mg to 10 mg / day / cm 2 The range may be:

[0025] The composition of the present invention may be used in the form of a composition to be applied to the skin. That is, the heat-treated cells of the Lactobacillus strain HJ1 (jojoba) may be used as is or in combination with other raw materials or base materials for formulations to form a composition for the skin. Examples of such compositions for the skin include lotions (skin lotions), cosmetic creams, milky lotions, skin milk, under-makeup, foundations, gels, jellies, powders, aerosols, bath additives, body shampoos, body rinses, soaps, cleansing foams, packs, lipsticks, lip balms, sun care products, ointments, patches, bandages, dressings, wound dressings, and the like. The use of such compositions for the skin includes not only those for humans but also those for animals.

[0026] On the other hand, the composition according to the present invention may be used in the form of an additive material to be added to such a skin composition.

[0027] The composition of the present invention may be used in the form of a composition to be applied to the oral cavity. That is, the heat-treated cells of the Lactobacillus casei HJ1 strain (joint acid bacteria) may be used as is or may be combined with other raw materials or base materials for formulations to form a composition for the oral cavity. Such oral compositions may be in the form of tablets, granules, troches, capsules, powders, drinks, jellies, gummies, candies, etc., and may be held in the mouth for a predetermined time and then swallowed. In addition, the composition may be in the form of mouthwash, chewing gum, toothpaste, etc., and may be not swallowed but simply held in the mouth. The composition may be in the form of an ointment, cream, spray, gel, lotion, etc., and used for application in the oral cavity. The form of use of such oral compositions includes not only humans but also animals.

[0028] On the other hand, the composition of the present invention may be used in the form of an additive material to be added to such an oral composition.

[0029] The composition of the present invention may be used for treating the inside of the mouth in the form of an animal feed product composition. That is, the heat-treated cells of the Lactobacillus casei HJ1 strain (jojoba) may be made into the form of an animal feed product composition as it is or in combination with other raw materials for animal feed products. Examples of animal feed products include, but are not limited to, feed for livestock, racehorses, ornamental animals, and pet foods.

[0030] On the other hand, the composition of the present invention may be used in the form of an additive material for animal feed products, as an additive to such animal feed products for oral use.

[0031] Furthermore, the composition according to the present invention may contain additives generally accepted for cosmetics, medicines, and foods, if necessary. Examples of additives include surfactants, colorants, preservatives, thickeners, pH adjusters, emulsifiers, etc. Also included are various carbohydrates, sweeteners, acidulants, emulsifiers, fruit juice, flavors (fragrances: peppermint oil, peppermint oil, etc.). More specifically, examples of additives include sugars such as sucrose, glucose, fructose, and honey, preservatives such as glyceryl caprylate, glyceryl undecylenate, and glyceryl ricinoleate, thickeners such as hydrated silica, cellulose gum, xanthan gum, and gellan gum, pH adjusters such as sodium citrate, calcium lactate, arginine, and citric acid, sugar alcohols such as sorbitol, xylitol, lactitol, erythritol, and palatinit, and emulsifiers such as lecithin, sucrose fatty acid esters, and glycerin sugar fatty acid esters. Other ingredients include various vitamins such as vitamin A, B vitamins, vitamin C, and vitamin E, herbal extracts, vegetable ingredients, milk ingredients, grain ingredients, cellulose, calcium stearate, and modified starch.

[0032] In the composition of the present invention, the content of heat-treated cells of the lactic acid bacteria HJ1 strain (joint acid bacteria) in various forms may be appropriately determined taking into consideration the relationship between the amount used and the effective dose. Typically, the content of heat-treated cells of the lactic acid bacteria HJ1 strain (joint acid bacteria) in terms of dry matter may be in the range of 0.001 to 100% by mass, 0.005 to 50% by mass, or 0.001 to 10% by mass. In addition, the content converted into the number of cells may be 2.5×10 7 ~2.5×10 12 cells / g, and may range from 1.25×10 8 ~1.25×10 12 cells / g, and may range from 2.5×10 8 ~2.5×10 11 The cell density may range from 100 to 1000 cells / g. EXAMPLES

[0033] The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to the scope of these examples.

[0034] (Preparation Example 1) Lactic acid bacteria HJ1 strain (lactic acid bacteria) was inoculated into MRS liquid medium and cultured at 37°C under aeration and agitation conditions. The culture was stopped at the late logarithmic growth phase, and the culture vessel was autoclaved at 121°C for 15 minutes for heat sterilization, and the resulting culture solution was used in the following tests. In the figure, it is called "HJ1 culture solution." The bacterial mass in the culture solution was 5 x 10 in terms of dry solids. 12 cells / g.

[0035] (Preparation Example 2) The cells were collected by centrifugation from the heat-sterilized culture solution obtained in Preparation Example 1, suspended in purified water, and used in the following tests. In the figure, this is called "HJ1 cells only." In the following tests, the amount of cells was used in combination with that of the other Preparation Examples.

[0036] (Preparation Example 3) The bacterial cells were collected by centrifugation from the heat-sterilized culture solution obtained in Preparation Example 1, and an equal amount of dextrin to the bacterial cells was added, followed by spray drying to prepare a powder product consisting of 50% lactic acid bacteria and 50% dextrin. In the figure, this is called "HJ1 product." In the following tests, the bacterial cell amounts used were the same as those of the other preparation examples.

[0037] <Test Example 1> The effect of Lactobacillus casei HJ1 strain (a zymobacterium) on the growth of Staphylococcus aureus was investigated.

[0038] Specifically, the samples prepared in Preparation Examples 1 to 3 were added to a Staphylococcus aureus growth medium (MRS medium) at 1.25 × 10 10 The cells were added to a concentration of 100000000000 cells / mL and cultured for 24 hours at 30°C under aeration and agitation conditions. After culture, a portion of the culture was spread on an agar medium containing a growth medium, and the number of viable bacteria contained in the culture after culture was calculated from the number of colonies that appeared.

[0039] As a result, as shown in Figure 1, compared to the group in which Staphylococcus aureus (S. aureus) was cultured alone, the number of live Staphylococcus aureus (S. aureus) bacteria was significantly reduced in the group in which the culture medium was added with the culture solution (heat-sterilized) of Lactobacillus HJ1 strain (joint acid bacteria) prepared in Preparation Example 1, the group in which the culture medium was added with the bacterial cells prepared in Preparation Example 2 (containing heat-sterilized Lactobacillus HJ1 strain (joint acid bacteria)), or the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized Lactobacillus HJ1 strain (joint acid bacteria)) was added to the medium.

[0040] <Test Example 2> We investigated the effect of Lactobacillus casei HJ1 strain (a zymobacterium) on the proliferation of Staphylococcus pseudintermedius, a causative bacterium of pyoderma.

[0041] Specifically, the samples prepared in Preparation Examples 1 to 3 were added to a growth medium for Pseudointermedius (casein peptone + NaCl medium) at 1.25 × 10 10 The cells were added to a concentration of 10 ...

[0042] As a result, as shown in Figure 2, compared to the group in which S. pseudointermedius was cultured alone, the number of live Staphylococcus aureus bacteria (S. aureus) was significantly reduced in the groups in which the culture medium was added with the culture solution (heat-sterilized) of the lactic acid bacteria HJ1 strain prepared in Preparation Example 1, the bacterial cells (containing heat-sterilized lactic acid bacteria HJ1 strain) prepared in Preparation Example 2, or the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacteria HJ1 strain)

[0043] <Test Example 3> We investigated the effect of Lactobacillus casei HJ1 strain (a zymobacterium) on the proliferation of Streptococcus mutans, a bacterium that causes dental caries.

[0044] Specifically, the sample prepared in Preparation Example 3 was added to a growth medium for Streptococcus mutans (trypticase soy + yeast extract medium) at a concentration of 1.25 × 10 10 The cells were added to a concentration of 10 ...

[0045] As a result, as shown in Figure 3, the number of live mutans bacteria (S. mutans) was significantly reduced in the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacteria HJ1 strain (lactic acid bacteria)) was added to the medium, compared to the group in which mutans bacteria (S. mutans) was cultured alone.

[0046] <Test Example 4> We investigated the effect of Lactobacillus casei HJ1 strain (a zymobacterium) on the proliferation of Staphylococcus epidermidis, a common skin flora.

[0047] Specifically, the samples prepared in Preparation Examples 2 and 3 were added to a growth medium for Staphylococcus epidermidis (MRS medium) at a concentration of 1.25 × 10 10 The cells were added to a concentration of 10 ...

[0048] As a result, as shown in Figure 4, compared to the group in which S. epidermis was cultured alone, no significant change was observed in the viable cell count of S. epidermis in the group in which the bacterial cells prepared in Preparation Example 2 (containing heat-sterilized Lactobacillus HJ1 strain (lactic acid bacteria)) or the powder product prepared in Preparation Example 3 (containing heat-sterilized Lactobacillus HJ1 strain) was added to the medium.

[0049] <Test Example 5> The odor reducing effect was investigated when the sample prepared in Preparation Example 3 was applied to the human oral cavity.

[0050] Specifically, 8.58 x 10 10 A solution of Lactobacillus casei strain HJ1 (lactic acid bacteria) prepared with purified water to a concentration of 10 cells / 10 mL was placed in the mouth, and after 5 minutes, measurements were started using a portable odor sensor (XP-329m, New Cosmos Electric Co., Ltd.). The odor measurement was performed by first sucking clean air into the sensor to return the displayed value to the base value, then inserting the sensor's intake nozzle about 1 cm into the mouth while holding breath, and sampling odor measurements over time.

[0051] As a result, as shown in FIG. 5, it was revealed that application of the lactic acid bacteria HJ1 strain (zygote) to the human oral cavity has the effect of reducing the odor.

[0052] <Test Example 6> The odor reducing effect was examined when the sample prepared in Preparation Example 3 was applied to human skin.

[0053] Specifically, 8.58 x 10 10 Lactobacillus casei strain HJ1 (lactic acid bacteria) was applied to the area behind the human ear at a concentration of 1.5 cells / mL. After application, odor measurements were performed using a portable odor sensor ("XP-329m", New Cosmos Electric Co., Ltd.). The odor measurements were performed in the same manner as in Test Example 5, except that the suction port of the sensor was placed a few millimeters above the application area.

[0054] As a result, as shown in FIG. 6, it was revealed that application of lactic acid bacteria HJ1 strain (zygote) to human skin has the effect of reducing the odor.

[0055] <Test Example 7> The odor reducing effect was examined when the sample prepared in Preparation Example 3 was applied to the oral cavity of a dog.

[0056] Specifically, 5×10 10 Lactic acid bacteria HJ1 strain (lactic acid bacteria) from Cells was applied directly to the gums, and after 5 minutes, the suction nozzle of a portable odor sensor ("XP-329m", New Cosmos Electric Co., Ltd.) was fixed 1 cm from the tip of the animal's mouth. Odor measurements were performed in the same manner as in Test Example 5.

[0057] As a result, as shown in FIG. 7, it was revealed that application of the lactic acid bacteria HJ1 strain (zygote) to the oral cavity of a dog had the effect of reducing the odor.

[0058] <Test Example 8> The odor reducing effect was examined when the sample prepared in Preparation Example 3 was applied to the oral cavity of a cat.

[0059] Specifically, 5×10 10 Lactic acid bacteria HJ1 strain (lactic acid bacteria) from Cells was applied directly to the gums, and after 5 minutes, the suction nozzle of a portable odor sensor ("XP-329m", New Cosmos Electric Co., Ltd.) was fixed 1 cm from the tip of the animal's mouth. Odor measurements were performed in the same manner as in Test Example 5.

[0060] As a result, as shown in FIG. 8, it was revealed that application of the lactic acid bacteria HJ1 strain (zygote) to the oral cavity of a cat had the effect of reducing the odor.

[0061] <Test Example 9> The immune activation ability of Lactobacillus casei HJ1 strain (a zymobacterium) was compared with other Lactobacillus plantarum strains by measuring the amount of IL-12 produced by mouse spleen cells. Specifically, the test was performed as follows.

[0062] Spleens were collected from BALB / cA mice (female, 10 weeks old). Cells were collected from the spleen using a cell strainer (100 μm pores) to obtain a cell concentration of 2.5 × 10 6 The cells were prepared in RPMI-1640 (Fujifilm Wako Pure Chemical) liquid medium containing 10% FBS (Thermo Fisher Scientific) and an appropriate amount of penicillin / streptomycin / neomycin (PSN) antibiotic mixture (Thermo Fisher Scientific) to a final concentration of 1.0 μg / mL. Each strain of L. plantarum, including HJ1 lactobacillus, was added to the cell solution to a final concentration of 1.0 μg / mL, and the cells were cultured at 37°C and 5% CO2 for 24 hours. The cytokine concentrations in the supernatant were then measured by ELISA and compared with the specific activity against the IL-12 production by OK432 (Picibanil, Chugai Pharmaceutical), an inactivated bacterial preparation used as a positive control.

[0063] As a result, as shown in Figure 9, the Lactobacillus casei HJ1 strain (a zymotryptic acid bacterium) showed higher immune activation ability than other Lactobacillus plantarum strains in a test for measuring IL-12 production levels using mouse spleen cells.

[0064] <Test Example 10> We investigated the effect of Lactobacillus casei HJ1 (a zymobacterium) on the proliferation of Porphyromonas macacae, a bacterium that causes periodontal disease.

[0065] Specifically, the sample prepared in Preparation Example 3 was diluted with 1.4 × 10 7 The cells were added to a concentration of 10 ...

[0066] As a result, as shown in Figure 10, the number of live Porphyromonas macacae bacteria (P. macacae) tended to decrease in the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacteria HJ1 strain (lactic acid bacteria)) was added to the medium, compared to the group in which Porphyromonas macacae was cultured alone.

[0067] <Test Example 11> We investigated the effect of Lactobacillus casei HJ1 strain (a zymobacterium) on the proliferation of Candida albicans, the causative agent of candidiasis.

[0068] Specifically, the sample prepared in Preparation Example 3 was diluted with 2.9 × 10 5 The cells were added to a concentration of 100000000000 cells / mL and cultured for 24 hours at 30°C under aeration and agitation conditions. After culture, a portion of the culture was spread on an agar medium containing a growth medium, and the number of viable bacteria contained in the culture after culture was calculated from the number of colonies that appeared.

[0069] As a result, as shown in Figure 11, the number of live Candida fungus (C. albicans) bacteria was significantly reduced in the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacteria HJ1 strain) was added to the medium, compared to the group in which Candida fungus (C. albicans) was cultured alone.

[0070] <Test Example 12> We investigated the effect of Lactobacillus casei HJ1 strain (a zymogen) on the proliferation of Malassezia furfur, the causative agent of tinea versicolor and seborrheic dermatitis.

[0071] Specifically, the sample prepared in Preparation Example 3 was soaked in a growth medium for M. furfur (malt extract 3.6% by mass, Tween 80 1% by mass, casein peptone 0.6% by mass, glycerin 1% by mass, olive oil 1% by mass) at 3.6 × 10 3 The cells were added to a concentration of 100000000000 cells / mL and cultured for 24 hours at 30°C under aeration and agitation conditions. After culture, a portion of the culture was spread on an agar medium containing a growth medium, and the number of viable bacteria contained in the culture after culture was calculated from the number of colonies that appeared.

[0072] As a result, as shown in Figure 12, the number of live M. furfur bacteria was significantly reduced in the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacteria HJ1 strain) was added to the medium, compared to the group in which M. furfur was cultured alone.

[0073] <Test Example 13> We investigated the effect of Lactobacillus casei HJ1 strain (a lactic acid bacterium) on the proliferation of Helicobacter pylori, a bacterium that causes gastritis and peptic ulcers.

[0074] Specifically, the sample prepared in Preparation Example 3 was added to a growth medium for Helicobacter pylori (BHI medium 3.7% by mass, FBS 10% by mass) at 1.88 × 10 5 The cells were added to a concentration of 10 ...

[0075] As a result, as shown in Figure 13, the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacteria HJ1 strain (lactic acid bacteria)) was added to the medium showed a significant decrease in the number of live H. pylori bacteria, compared to the group in which H. pylori was cultured alone.

[0076] [summary] According to the results of Test Examples 1 to 4 and 10 to 13, it was revealed that while the lactic acid bacteria HJ1 strain (lactic acid bacteria) does not show much inhibitory effect on the growth of Staphylococcus epidermidis, which is known as a beneficial bacterium of the skin, it has an inhibitory effect on the growth of harmful bacteria such as Staphylococcus aureus, Bacillus pseudointermedius which is the causative bacterium of pyoderma, Streptococcus mutans which is the causative bacterium of dental caries, Porphyromonas maccae which is the causative bacterium of periodontal disease, Candida fungi which are the causative bacteria of candidiasis, M. versicolor which is the causative bacterium of tinea versicolor and seborrheic dermatitis, and Helicobacter pylori which is the causative bacterium of gastritis and peptic ulcers.

[0077] Furthermore, the results of Test Examples 5 to 8 revealed that the lactic acid bacteria HJ1 strain (zygote) had the effect of reducing odors in the oral cavity and on the skin.

[0078] Furthermore, according to the results of Test Example 9, the lactic acid bacteria HJ1 strain (nucleotidic acid bacteria) was confirmed to have high immune activation ability in a test for measuring the amount of IL-12 produced using mouse spleen cells.

Claims

1. A composition for inhibiting harmful bacteria on the skin and in the oral cavity, which contains heat-treated cells of Lactiplantibacillus plantarum HJ1 (lactic acid bacteria HJ1 strain (lactic acid bacteria)) as an active ingredient and is applied to the skin or the oral cavity.

2. The composition for inhibiting harmful bacteria on the skin and in the oral cavity according to claim 1, wherein the harmful bacteria is one or more selected from the group consisting of Staphylococcus aureus, Staphylococcus pseudointermedius, Streptococcus mutans, Porphyromonas macacae, Candida albicans, Malassezia furfur, and Helicobacter pylori.

3. 2. The composition for inhibiting harmful bacteria on the skin and oral cavity according to claim 1, which has the effect of reducing odor on the skin or oral cavity to which it is applied.

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