Bifidobacterium stercoris strain and uses thereof

Abstract

The present application relates to a Bifidobacterium stercoris strain and uses thereof.

Description

Bifidobacterium stuccoris strain and its uses

[0001] The present application relates to a strain of Bifidobacterium stuccoris and its uses.

[0002] A microbiome refers to the sum of microbial genomes existing within a specific environment. Approximately 100 trillion microorganisms inhabit the human body, forming an ecosystem where they interact with one another. Furthermore, the microbiome influences the host's immunity and health, and is associated with the development of diseases.

[0003] Although the pathogenesis of diarrhea-predominant irritable bowel syndrome (IBS-d) has not yet been fully elucidated, an excessive serotonin response is a representative mechanism; it is known that serotonin stimulates enteric nerves, causing the intestines to become sensitized and leading to excessively active bowel movements. In fact, drugs that regulate serotonin signaling, such as alosetron, have received FDA approval and are commercially available. However, existing drugs are less effective for patients with severe IBS-d symptoms and cause side effects such as ischemic colitis and severe constipation, so there are still unmet needs for IBS-d treatments.

[0004] Inflammatory responses are known as another pathogenic mechanism of IBS-d. Mast cells are identified as a representative associated immune cell that induces IBS-d pathology by releasing substances such as histamine and trypsinase through degranulation. In addition, pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6, or low-grade inflammation, also influence IBS-d pathology. However, there are currently no therapeutic agents for IBS-d that target these related mechanisms.

[0005] Against this backdrop, one objective of the present application is to provide a novel strain with excellent irritable bowel syndrome alleviation effects based on serotonin reduction effects, mast cell degranulation inhibition effects, and various uses thereof.

[0006] Another objective of the present application is to provide a composition, food, probiotic, or quasi-drug comprising one or more selected from the group consisting of a strain according to one example of the present application, a culture of said strain, a lysed product of said strain, and an extract of said strain.

[0007] One example of the present application relates to the Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP.

[0008] Another example of the present application relates to a composition, food, probiotic, or quasi-drug comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0009] Another example of the present application relates to a method for reducing the serotonin level of a subject, comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0010] Another example of the present application relates to a method for preventing or treating serotonin syndrome, comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0011] Another example of the present application relates to a method for preventing or treating irritable bowel syndrome, comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0012] Another example of the present application relates to a method for preventing or treating mast cell activation syndrome (MCAS), comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0013] Another example of the present application relates to a method for preventing or treating inflammatory bowel disease, comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0014] Another example of the present application relates to a method for strengthening tight junctions, comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0015] Another example of the present application relates to a method for increasing the relative abundance of Butyrimonas strains in the intestine, comprising the step of administering to a subject one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0016]

[0017] The present invention will be described in more detail below.

[0018] A Bifidobacterium stercoris strain KC84 having deposit number KCTC15975BP according to one example of the present application may have the 16S rRNA sequence of SEQ ID NO. 1.

[0019] The strain may have one or more features selected from the group consisting of (1) to (6) below:

[0020] (1) Inhibits the secretion of serotonin;

[0021] (2) Promotes the reabsorption of serotonin;

[0022] (3) Inhibits the synthesis of serotonin;

[0023] (4) Strengthening of tight junctions of colon mucosal epithelial cells;

[0024] (5) Inhibition of degranulation of mast cells; and

[0025] (6) Increase the relative abundance of Butyrimonas strains in the intestines.

[0026] The strain according to one example of the present application has a therapeutic effect on IBS-d by inhibiting serotonin secretion and mast cell degranulation, which are the pathogenic mechanisms of IBS-d, and the excellent therapeutic efficacy of the strain according to one example of the present application was verified in an IBS-d animal model.

[0027] Specifically, in the embodiment of the present application, the serotonin secretion inhibitory ability was verified by treating enterochromaffin cells with a strain according to one example of the present application. As a result, based on 100% of the serotonin secretion of the untreated control group, the serotonin secretion of the enterochromaffin cells was inhibited to 10% or less, showing a significantly superior serotonin secretion inhibitory effect. This was a significantly superior serotonin secretion inhibitory effect compared to telotristat ethyl, which is known as a conventional serotonin inhibitor.

[0028] Specifically, in the embodiments of the present application, the mast cell degranulation inhibitory effect of the strain according to one example of the present application was verified, and based on the mast cell degranulation level of 100% of the untreated control group, the mast cell degranulation level was found to be 50% or less, 45% or less, or 42% or less, showing a significantly superior mast cell degranulation inhibitory effect. This was a significantly superior mast cell degranulation inhibitory effect compared to the Bifidobacterium stuccoris KC109 strain, which belongs to the same genus as the strain according to one example of the present application.

[0029] In addition, the excellent efficacy of the strain according to one example of the present application was verified in a 5-HTP mouse model, an animal model accompanied by severe diarrhea, and it was confirmed that it has a therapeutic effect even on severe irritable bowel syndrome. Specifically, in a 5-HTP animal model with severe diarrhea, administration of the strain according to one example of the present application not only improved the phenotype but also further verified a decrease in serotonin levels, changes in gene expression, and histological changes.

[0030] More specifically, in the embodiment of the present application, the serotonin-reducing effect of the strain according to one example of the present application was verified in the colon tissue of a 5-HTP mouse model, and based on 100% of the serotonin-positive area of ​​the untreated control group, the serotonin-positive area was found to be 60% or less, 55% or less, 50% or less, or 45% or less, showing a significantly superior serotonin-reducing effect.

[0031] More specifically, in the embodiment of the present application, the effect of reducing bowel frequency of the strain according to one example of the present application was verified in a 5-HTP mouse model, and based on the bowel frequency of the untreated control group at 100%, the bowel frequency was found to be 80% or less or 75% or less, showing a significantly superior effect of reducing bowel frequency.

[0032] More specifically, in the embodiment of the present application, the effect of improving diarrhea scores of the strain according to one example of the present application was verified in a 5-HTP mouse model, and based on the diarrhea score of the untreated control group at 100%, the diarrhea score was found to be 70% or less, 65% or less, or 60% or less, showing a significantly superior effect in improving diarrhea scores.

[0033] More specifically, in the embodiment of the present application, the effect of the strain according to one example of the present application on reducing fecal moisture content was verified in a 5-HTP mouse model, and the fecal moisture content was statistically significantly reduced compared to the untreated control group (p<0.05), showing an excellent effect on reducing fecal moisture.

[0034] More specifically, in the embodiment of the present application, the colitis improvement effect of the strain according to one example of the present application was verified in a 5-HTP mouse model, and based on the colitis index of the untreated control group at 100%, the colitis index was found to be 80% or less, 75% or less, 70% or less, or 65% or less, showing a significantly superior colitis improvement effect.

[0035] More specifically, in the embodiments of the present invention, the effect of gene expression change by the strain according to one example of the present application was verified, and as a result, the expression level of the serotonin transporter gene increased and the expression level of the tryptophan hydrolase gene decreased, thereby verifying the effect of increased serotonin reabsorption and decreased biosynthesis.

[0036] In addition, phenotypic improvement and changes in microbiome members were verified by administering the strain according to one example of the present application in a zymosan animal model characterized by inflammation as a TLR2 agonist. Specifically, the preventive and therapeutic effects of irritable bowel syndrome and the effect of increasing the relative abundance of butyric acid-producing strains in the intestines were verified by administering the strain according to one example of the present application in a zymosan animal model.

[0037] More specifically, in the embodiments of the present application, the effect of the strain according to one example of the present application on reducing fecal moisture content was verified in a zymosan animal model. As a result, the fecal moisture content was statistically significantly reduced (p<0.01) compared to the untreated control group, demonstrating an excellent effect on reducing fecal moisture.

[0038] More specifically, in the embodiments of the present application, the effect of the strain according to one example of the present application on changes in intestinal microbiome members was verified in a zymosan animal model. As a result, the relative abundance of Butyrimonas strains that produce butyrate increased in the intestines, demonstrating an excellent effect on improving the intestinal microbiome.

[0039] Accordingly, another example of the present application relates to a composition for increasing the relative abundance of Butyrimonas strains in the intestine, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0040] In addition, phenotypic improvement and a reduction in serotonin levels were verified in a WAS rat animal model, which is a model that induces IBS-d through mental stress, by administering the strain according to one example of the present application. Specifically, in a WAS rat animal model, the preventive and therapeutic effects of irritable bowel syndrome and the effect of reducing serotonin concentration in the lumen contents were verified by administering the strain according to one example of the present application.

[0041] More specifically, in the embodiments of the present application, the effect of the strain according to one example of the present application on improving diarrhea scores was verified in a WAS rat animal model. Based on a diarrhea score of 100% of the untreated control group, the diarrhea score was found to be 30% or less, 25% or less, 20% or less, or 15% or less, demonstrating a significantly superior effect in preventing and treating irritable bowel syndrome. This effect was significantly superior to that of the positive control, alosetron.

[0042] More specifically, in the embodiments of the present application, the effect of reducing bowel frequency of the strain according to one example of the present application was verified in a WAS rat animal model. Based on 100% of the fecal pellet output (FPO) in the feces obtained from the untreated control group, the FPO in the feces obtained was found to be 30% or less, 25% or less, or 20% or less, showing a significantly superior effect in preventing and treating irritable bowel syndrome. This effect was significantly superior to that of the positive control, alosetron.

[0043] More specifically, in the embodiments of the present application, the effect of the strain according to one example of the present application on reducing bowel frequency was verified in a WAS rat animal model. Based on the average FPO (mean fecal pellet output) of 100% of the untreated control group, the average FPO was found to be 80% or less, 75% or less, 70% or less, 65% or less, or 60% or less, demonstrating a significantly superior effect in preventing and treating irritable bowel syndrome. This effect was significantly superior to that of the positive control, alosetron.

[0044] More specifically, in the embodiment of the present application, the effect of reducing serotonin concentration in the luminal content of the strain according to one example of the present application was verified in a WAS rat animal model, and based on 100% of the serotonin concentration in the luminal content of the untreated control group, the serotonin concentration in the luminal content was found to be 85% or less, 80% or less, or 77% or less, showing an excellent serotonin reduction effect.

[0045] In addition, the strain according to one example of the present application is derived from the human body, does not exhibit cytotoxicity, and has very little safety concern as its antibiotic resistance is below the EFSA standard, so the possibility of side effects is low.

[0046] Accordingly, it has been verified that the strain according to one example of the present application alleviates indicators and symptoms associated with irritable bowel syndrome.

[0047] Another example of the present application relates to a pharmaceutical composition for the prevention or treatment of serotonin syndrome, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0048] The above serotonin syndrome is caused by an excessive accumulation of serotonin in the body and may be caused by drugs or substances that increase serotonin levels. The above serotonin syndrome may exhibit various symptoms ranging from mild to severe, and may be accompanied, for example, by neurological symptoms, autonomic nervous system symptoms, and / or muscular symptoms. The above neurological symptoms may include anxiety, seizures, and / or headaches, the above autonomic nervous system symptoms may include high fever, sweating, high blood pressure, vomiting, and / or diarrhea, and the above muscular symptoms may include, but are not limited to, muscle spasms and / or muscle rigidity.

[0049] The above composition may inhibit serotonin secretion of a subject. Specifically, the above composition may inhibit serotonin secretion of enterochromaffin cells.

[0050] The above composition may promote serotonin reabsorption of the subject.

[0051] The above composition may inhibit serotonin synthesis of the subject.

[0052] The above composition may reduce the serotonin concentration of the luminal contents of a subject. Specifically, in the embodiments of the present application, the serotonin reduction effect on the luminal contents of a strain according to one example of the present application was verified. As a result, based on a serotonin concentration of 100% of the luminal contents of the untreated control group, the serotonin concentration of the luminal contents was found to be 90% or less, 85% or less, or 80% or less, demonstrating an excellent serotonin reduction effect. On the other hand, no statistically significant reduction was observed in the positive control group treated with ondansetron.

[0053] The above composition may inhibit the movement of serotonin from the blood to the intestines in the subject. Specifically, in the embodiments of the present application, the ratio of serotonin that moved from the serum to the lumen of a strain according to one example of the present application was measured. Based on a serotonin movement ratio of 100% for the untreated control group, the serotonin movement ratio was found to be 90% or less, 85% or less, or 80% or less, demonstrating an excellent inhibitory effect on serotonin movement. On the other hand, no statistically significant decrease was observed in the positive control group treated with ondansetron.

[0054] The above composition may inhibit the movement of serotonin from the colon to the lumen in the subject. Specifically, in the embodiments of the present application, the ratio of serotonin moving from the colon to the lumen of the strain according to one example of the present application was measured, and based on the serotonin movement ratio of 100% of the untreated control group, the serotonin movement ratio was found to be 70% or less, 65% or less, or 60% or less, showing an excellent inhibitory effect on serotonin movement. On the other hand, no statistically significant decrease was observed in the positive control group treated with ondansetron.

[0055] Another example of the present application relates to a pharmaceutical composition for the prevention or treatment of irritable bowel syndrome, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain. In one example, said irritable bowel syndrome may be diarrhea-predominant irritable bowel syndrome.

[0056] The above-mentioned irritable bowel syndrome is a chronic digestive disorder characterized by abdominal pain and abnormal bowel movements, primarily caused by functional problems in the intestines. Specifically, abdominal pain occurs due to intestinal dysfunction, and diarrheal irritable bowel syndrome may be accompanied by frequent bowel movements and diarrhea. Irritable bowel syndrome can occur when the intestinal nerves are stimulated by excessive serotonin. Accordingly, a pharmaceutical composition according to one example of the present application can lower serotonin levels and exhibit a preventive or therapeutic effect for irritable bowel syndrome.

[0057] The above composition may reduce the moisture content of stool and / or the frequency of bowel movements.

[0058] Another example of the present application relates to a pharmaceutical composition for the prevention or treatment of mast cell activation syndrome (MCAS), comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0059] The above-mentioned mast cell activation syndrome is a disease caused by mast cells becoming excessively activated and secreting inflammatory substances.

[0060] The above mast cell activation syndrome may include, but is not limited to, one or more selected from the group consisting of allergies, urticaria, rashes, asthma, and chronic inflammation.

[0061] Another example of the present application relates to a composition for the prevention or treatment of inflammatory bowel disease comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0062] In the embodiments of the present application, the effect of reducing intestinal inflammation by the strain according to one example of the present application was verified. As a result, the expression of inflammatory cytokines in colon tissue was reduced by the administration of the strain according to one example of the present application, and the relative abundance in the intestine of the strain producing butyric acid, which has preventive and therapeutic effects for inflammatory bowel disease, was increased, thereby verifying the anti-inflammatory effect, specifically the anti-inflammatory effect in the intestine.

[0063] Another example of the present application relates to a composition for strengthening tight junctions of colon mucosal epithelial cells, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0064] In the embodiment of the present application, the effect of strengthening tight junctions of the strain according to one example of the present application was verified. As a result, the expression amount of tight junction genes increased upon administration of the strain according to one example of the present application, thereby verifying the effect of strengthening tight junctions of colon mucosal epithelial cells.

[0065] Another example of the present application relates to a food comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

[0066] The above food may be in the form of, for example, meat, bread, chocolate, candies, jellies, snacks, confectionery, kimchi, sauces, cheese, dairy products, powders, beverages, or vitamin complexes. The above food may be a health functional food.

[0067] The above food may include, as an active ingredient, ingredients that are typically added during food manufacturing. The above added ingredients include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavorings. The carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, oligosaccharides, etc.), and polysaccharides (e.g., conventional sugars such as dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavorings, natural flavorings (taumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavorings (saccharin, aspartame, etc.) may be used.

[0068] For example, when a food product according to one example of the present application is manufactured as a drink, it may additionally include citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, jujube extract, or licorice extract.

[0069] Food according to one example of the present application includes processed forms of all natural materials, such as functional food, nutritional supplement, health food, and food additives. Foods of the above types may be manufactured in various forms according to conventional methods known in the art. For example, as a health food, the strain itself described above may be manufactured in the form of tea, juice, and drink for consumption, or may be consumed after granulation, encapsulation, and powdering. In addition, food products may be in the form of beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruit, bottled fruit, jam, marmalade, etc.), fish, meat and its processed foods (e.g., ham, sausage, corned beef, etc.), bread and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, malt syrup, dairy products (e.g., yogurt, fermented milk, butter, cheese, etc.), edible vegetable oils and fats, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.). Furthermore, in order to use the strain according to one example of the present application, the culture of the strain, the crushed material of the strain, and the extract of the strain, etc., as food additives, they may be prepared and used in the form of a powder or a concentrate.

[0070] Another example of the present application relates to a probiotic comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain. said probiotic may be in the form of a powder, granule, tablet, or capsule.

[0071] Another example of the present application relates to a quasi-drug comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain. The quasi-drug composition may be in a form selected from the group consisting of toothpaste, mouthwash, oral spray, oral ointment, mouthwash, mouthwash, bandage, and insect (mosquito, tick, etc.) repellent.

[0072] In this application, the term "prevention" means suppressing or delaying the onset of a disease, disorder, or condition. Prevention may be considered complete if the onset of a disease, disorder, or condition is suppressed or delayed for a scheduled period.

[0073] In this application, the term "treatment" means partially or completely alleviating, improving, mitigating, inhibiting, or delaying symptoms associated with a specific disease, disorder and / or condition or condition, reducing severity, or reducing the occurrence of one or more symptoms or features.

[0074] In this application, the term "strain" refers to a collection of individuals having homogeneous genetic characteristics that have proliferated through asexual reproduction originating from a single cell, and may be live strains, dead strains, or heat-inactivated strains.

[0075] In this application, the term "culture of a strain" refers to a product obtained after culturing a strain according to one example of this application. The culture may be the whole culture of the strain according to one example of this application, a diluted solution, a concentrate, a dried product, a freeze-dried product, a crushed product, and / or a fraction thereof. The concentrate may be obtained by centrifuging or evaporating the culture; the dried product may be obtained by drying the culture using a dryer or the like; the freeze-dried product may be obtained by freeze-drying the culture using a freeze-dryer or the like; the crushed product may be obtained by physically or ultrasonically treating the strain or culture; and the fraction may be obtained by applying the culture, crushed product, etc., to methods such as centrifugation or chromatography. The culture may be in a solid phase (solid, e.g., dried product), a liquid phase (liquid), or a fluid phase, but is not limited thereto. In one example, the culture may refer to the entire medium containing the cultured strain obtained by culturing the strain according to one example of the present application for a certain period, its metabolites, and / or excess nutrients. In one example, the culture may be one in which the strain according to one example of the present application has been removed or has not been removed. In one example, the culture may refer to the remaining components excluding the strain (cells) from a culture obtained by culturing the strain according to one example of the present application in a medium. In one example, the culture may be a culture medium (or culture) from which the strain (cells) has been removed from a culture medium in which the strain according to one example of the present application has been cultured in a medium. The culture medium (or culture) from which the strain has been removed may be a cell-free culture medium (or culture) or a culture medium containing dead cells, and may be, for example, a filtrate (supernatant after centrifugation) from which the strain has been removed by filtration or centrifugation, and / or a culture medium (or dried culture medium) containing dead cells.Specifically, the culture may exhibit anti-inflammatory activity at a level equivalent to the activity exhibited by the strain according to one example of the present application, or preventive, improving, or therapeutic activity for inflammatory diseases.

[0076] In this application, the term "crushed material of a strain" may refer to a product obtained by crushing a strain according to one example of this application by chemical or physical force. Specifically, said crushed material may exhibit anti-inflammatory activity at a level equivalent to the activity exhibited by the strain according to one example of this application, or preventive, remedial, or therapeutic activity for inflammatory diseases.

[0077] In this application, the term "extract" may refer to a product obtained by extracting a strain according to one example of this application, a culture of said strain, a lysate of said strain, or a mixture thereof, regardless of the extraction method, extraction solvent, extracted component, or form of the extract; it is a broad concept that includes all substances that may be obtained by processing or treating by other methods after extraction. For example, said extract may be an extract of said strain according to one example of this application, an extract of said strain's culture, or an extract of said strain's lysate. Specifically, said extract may exhibit anti-inflammatory activity or preventive, improving, or therapeutic activity of inflammatory diseases at a level equivalent to the activity exhibited by said strain, said strain's culture, or said strain's lysate according to one example of this application.

[0078] A composition, food, probiotic, quasi-drug, etc. according to one example of the present application may additionally include one or more active ingredients exhibiting the same or similar functions in addition to the above active ingredients.

[0079] In addition, compositions, foods, probiotics, quasi-drugs, etc. according to one example of the present application may be manufactured in a unit dose form or contained in a multi-dose container by formulating them using a pharmaceutically acceptable carrier according to a method that can be clearly carried out by a person with ordinary knowledge in the technical field to which the invention belongs. In the present application, the term "carrier" means a compound that facilitates the addition of a compound into a cell or tissue, and the term "pharmaceutically acceptable" means a composition that is physiologically acceptable and, when administered to humans, does not typically cause allergic reactions such as gastrointestinal disorders or dizziness, or similar reactions.

[0080] The above pharmaceutically acceptable carriers are those commonly used in formulations and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0081] In addition, the composition, food, probiotic, quasi-drug, etc. according to one example of the present application may additionally include additives such as fillers, anticoagulants, lubricants, humectants, fragrances, emulsifiers, and preservatives in addition to the above components. In the present application, the content of the additives is not particularly limited and can be appropriately adjusted within the content range used in conventional formulations.

[0082] In addition, the composition, food, probiotic, quasi-drug, etc. according to one example of the present application may be formulated into an oral preparation. Non-limiting examples of the oral preparation include tablets, troches, lozenges, water-soluble suspensions, oily suspensions, prepared powders, lyophilized-dried powders, spray-dried powders, granules, emulsions, capsules, hard capsules, soft capsules, enteric capsules, syrups, or elixirs. For example, the composition, food, probiotic, quasi-drug, etc. according to one example of the present application may include an enteric coating. To formulate a pharmaceutical composition or food composition according to one example of the present application for oral administration, binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose, or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; magnesium stearate, calcium stearate, sodium stearyl fumarate, etc. may be used, and sweeteners, flavorings, syrups, etc. may also be used. Furthermore, in the case of capsules, in addition to the materials mentioned above, liquid carriers such as fatty oils may be additionally used.

[0083] In this application, the term "excipient" refers to any substance that is not a therapeutic agent and is used as a carrier or medium for the delivery of a therapeutic agent or is added to a pharmaceutical composition. By doing so, handling and storage characteristics are improved, or the formation of a unit dose of the composition is allowed and facilitated.

[0084] A composition according to one example of the present application, e.g., a pharmaceutical composition, may be formulated and used in various forms according to conventional methods for each intended use, such as oral formulations like liquids, suspensions, powders, granules, tablets, capsules, pills, extracts, emulsions, syrups, and aerosols, and injectable formulations such as sterile injectable solutions. It may be administered orally or through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and local administration. In the present application, the term "oral administration" means administration to a substance prepared so that the active substance is digested, i.e., to the gastrointestinal tract for absorption.

[0085] The preferred dosage of a composition or food according to one example of the present application may vary depending on the patient's condition and weight, age, gender, health condition, dietary constitutional characteristics, properties of the preparation, degree of disease, time of administration of the composition, method of administration, duration or interval of administration, excretion rate and form of the drug, and may be appropriately selected by a person skilled in the art.

[0086] In this application, the term "effective dose" refers to an amount of a composition of an active ingredient sufficient to treat a specific symptom. This may vary depending on the formulation method of the pharmaceutical composition or food composition, the mode of administration, the time of administration and / or the route of administration, etc., and may vary depending on various factors and similar factors well known in the pharmaceutical field, including the type and degree of response to be achieved by administering the pharmaceutical composition or food composition, the type of individual to be administered, age, body weight, general health condition, symptoms or severity of disease, gender, diet, excretion, and the components of other compositions used simultaneously or at once with the individual. A person skilled in the art can easily determine and prescribe an effective dose for the intended treatment.

[0087] The composition or food according to one example of the present application may be administered once a day or divided into several doses. The composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Taking all of the above factors into consideration, it may be administered in an amount that obtains maximum effect with a minimum amount without side effects.

[0088] For example, a composition according to one example of the present application comprises 0.001 to 10,000 mg, 0.001 to 5,000 mg, 0.001 to 1,000 mg, 0.001 to 500 mg, 0.001 to 300 mg, 0.001 to 100 mg, 0.001 to 50 mg, 0.001 to 30 mg, 0.001 to 10 mg, 0.001 to 5 mg, 0.001 to 1 mg, 0.001 to 0.5 mg, 0.001 to 0.1 mg, 0.001 to 0.05 mg, 0.001 to 0.01 mg, 0.01 to 10,000 mg, 0.01 to 5,000 mg per 1 kg of body weight, 0.01 to 1,000 mg, 0.01 to 500 mg, 0.01 to 300 mg, 0.01 to 100 mg, 0.01 to 50 mg, 0.01 to 30 mg, 0.01 to 10 mg, 0.01 to 5 mg, 0.01 to 1 mg, 0.01 to 0.5 mg, 0.01 to 0.1 mg, 0.01 to 0.05 mg, 0.1 to 10,000 mg, 0.1 to 5,000 mg, 0.1 to 1,000 mg, 0.1 to 500 mg, 0.1 to 300 mg, 0.1 to 200 mg, 0.1 to 100 mg, 0.1 to 50 mg, 0.1 to 30 mg, 0.1 to 10 mg, 0.1 to 5 mg, 0.1 to 1 mg, 0.1 to 0.5 mg, 1 to 10,000 mg, 1 to 5,000 mg, 1 to 1,000 mg, 1 to 500 mg, 1 to 300 mg, 1 to 200 mg, 1 to 100 mg, 1 to 50 mg, 1 to 10 mg, 1 to 5 mg, 10 to 10,000 mg, 10 to 5,000 mg, 10 to 1,000 mg, 10 to 500 mg, 10 to 300 mg, 10 to 200 mg, 10 to 100 mg, 10 to 50 mg, 10 to 40 mg, 10 to 30 mg, 10 to 20 mg, 10 to 10,000 mg, 100 to 5,000 mg, 100 to It may be administered in daily doses of 1,000 mg, 100 to 500 mg, 100 to 300 mg, or 100 to 200 mg, but is not limited thereto. For example, the daily dose of the composition according to one example of the present application may be 0.001 to 10 g / day, 0.001 to 5 g / day, 0.01 to 10 g / day, or 0.01 to 5 g / day based on oral administration. Additionally, the total daily dose may be divided and administered continuously or discontinuously as needed.

[0089] The Bifidobacterium stercoris KC84 strain, the culture of the strain, the lysate of the strain, and the extract of the strain according to one example of the present application are safe as they are non-toxic to the human body and have the advantage of being easily accessible as therapeutic agents without negative perceptions. In addition, they exhibit excellent effects in improving IBS-d disease without the risk of side effects associated with conventional IBS-d disease treatments, making them highly useful for industrial applications.

[0090] FIG. 1 is a figure showing the serotonin inhibitory effect of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where *, **, and *** represent p < 0.05, p < 0.01, and p < 0.001, respectively, compared to the control group (student t-test).

[0091] FIG. 2 is a diagram showing the inhibitory effect of the Bifidobacterium stuccoris KC84 strain on mast cell degranulation according to one example of the present application, where * and *** represent p < 0.05 and p < 0.001, respectively, compared to a control group treated with the stimulator (A23187) (student t-test).

[0092] FIG. 3a is a figure showing that defecation frequency is significantly reduced by administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in an irritable bowel syndrome disease model, where * indicates p < 0.05 compared to a control group treated with 5-HTP (student t-test).

[0093] FIG. 3b is a figure showing that the diarrhea score is significantly alleviated by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in an irritable bowel syndrome disease model, where *, **, and *** represent p < 0.05, p < 0.01, and p < 0.001, respectively, compared to the control group treated with 5-HTP (student t-test).

[0094] FIG. 3c is a figure showing that the water content of stool is significantly reduced by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in an irritable bowel syndrome disease model, where * and *** represent p < 0.05 and p < 0.001, respectively, compared to a control group treated with 5-HTP (student t-test).

[0095] FIG. 3d is the result of histologically verifying the preventive and therapeutic effects of the Bifidobacterium stuccoris KC84 strain according to one example of the present application on irritable bowel syndrome, where ** and *** represent p < 0.01 and p < 0.001, respectively, compared to the control group treated with 5-HTP (student t-test).

[0096] FIG. 4a is a figure showing a decrease in serotonin in luminal content (p = 0.06) by administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where * and ** represent p < 0.05 and p < 0.01, respectively, compared to a control group treated with 5-HTP (student t-test).

[0097] FIG. 4b is a figure showing the ratio of the amount of serotonin in the luminal content to the amount of serotonin in the serum after administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where * and ** represent p < 0.05 and p < 0.01, respectively, compared to the control group treated with 5-HTP (student t-test).

[0098] FIG. 4c is a figure showing the ratio of the amount of serotonin in the luminal content to the amount of serotonin in the colon after administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where * and ** represent p < 0.05 and p < 0.01, respectively, compared to the control group treated with 5-HTP (student t-test).

[0099] FIG. 4d is the result of histologically verifying the serotonin-reducing effect of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where ** and *** represent p < 0.01 and p < 0.001, respectively, compared to the control group treated with 5-HTP (student t-test).

[0100] FIG. 5a is a figure showing the results of an increase in Slc6a4 gene expression and a decrease in Tph1 gene expression by the Bifidobacterium stuccoris KC84 strain according to an example of the present application, where *, **, and *** represent p < 0.05, p < 0.01, and p < 0.001, respectively, compared to a control group treated with 5-HTP (student t-test).

[0101] FIG. 5b is a figure showing the results of a decrease in the expression level of the IL-β gene by the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where * and ** indicate p < 0.05 and p < 0.01 compared to the control group treated with 5-HTP (student t-test).

[0102] FIG. 5c is a figure showing the result of increased expression of the Tjp1 gene by the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where *, *** represent p < 0.05 and p < 0.001 compared to the control group treated with 5-HTP (student t-test).

[0103] FIG. 6 is a figure showing that in a zymosan mouse model, the water content, an indicator of the degree of diarrhea, is significantly reduced by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, where ** and *** represent p < 0.01 and p < 0.001, respectively, compared to the control group treated with zymosan (student t-test).

[0104] FIG. 7a is a diagram showing the results of α-diversity analysis after administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application.

[0105] FIG. 7b is a diagram showing the results of β-diversity analysis after administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application.

[0106] FIG. 7c is a diagram showing the LEfSe analysis results after administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application.

[0107] FIG. 8a is a figure showing that the diarrhea score is significantly alleviated by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in a WAS rat model, where * and ** represent p < 0.05 and p < 0.01, respectively, compared to the control group administered WAS (student t-test).

[0108] FIG. 8b is a figure showing that the fecal pellet output of diarrhea score is significantly reduced by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in a WAS rat model, where * and ** represent p < 0.05 and p < 0.01, respectively, compared to the control group administered WAS (student t-test).

[0109] FIG. 8c is a figure showing that the mean fecal pellet output is significantly reduced by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in a WAS rat model, where *** indicates p < 0.001 compared to the control group administered WAS (student t-test).

[0110] FIG. 9 is a figure showing that serotonin in the luminal content was significantly reduced by administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application in a WAS rat model, where * indicates p < 0.05 compared to the control group administered WAS (student t-test).

[0111] FIG. 10 is a diagram verifying that the antibiotic resistance of the Bifidobacterium stuccoris KC84 strain according to one example of the present application is below the EFSA standard.

[0112] FIG. 11 is a diagram verifying that the Bifidobacterium stuccoris KC84 strain according to one example of the present application is a safe strain as it does not exhibit hemolytic activity.

[0113] FIG. 12 is a diagram verifying that the Bifidobacterium stuccoris KC84 strain according to one example of the present application is a safe strain as it does not exhibit cytotoxicity, and * and ** represent p < 0.05 and p < 0.01, respectively, compared to the Staphylococcus aureus treatment group at the same concentration (student t-test).

[0114] The present application will be explained in more detail below by means of the following examples. However, these examples are merely illustrative of the present application, and the scope of the present application is not limited by these examples.

[0115]

[0116] Example 1. Identification of Bifidobacterium stuccoris strains

[0117] Three candidate strains were isolated by inoculating fecal samples from healthy Korean women in their 40s onto BL agar medium and performing two subcultures at 48-hour intervals in an anaerobic environment at 37°C, followed by colony picking and pure culture. They were named KC84, KC109, and KC19, respectively.

[0118] Three candidate strains were subcultured and isolated by colony picking. The isolated strains were cultured in a 37°C CO2 incubator under an anaerobic environment for 48 hours, followed by centrifugation at 13,000 rpm at 4°C to obtain the strains. Next, 0.2 µl of the strain pellet was added to 25 µl of lysis buffer (pH 10; 2.5 M NaCl, 100 mM EDTA, 10 mM Trizma base, 1% Triton X-100) and heat-treated at 95°C for 10 minutes to obtain a template. Subsequently, the V4 region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) using a G-Taq PCR kit to obtain the PCR product. Primers 27F and 1492R were used. The PCR product was purified using an Ultra Clean PCR clean-up Kit (Mobio Laboratories Inc.), and the strains were identified through gene sequencing analysis. The 16S rRNA sequence of the KC84 strain is shown in Table 1.

[0119] By comparing the 16S rRNA sequence of strain KC84 with the NCBI Bacteria and Archaea: 16S ribosomal RNA project DB, strain KC84 was identified as Bifidobacterium stercoris. The strain was deposited with the Korean Collection for Type Culture at the Korea Research Institute of Biotechnology and Bioengineering (KRIBB), an international depositary institution under the Treaty of Budapest, and was assigned deposit number KCTC15975BP. Strain KC109 was identified as Bifidobacterium stercoris, identical to strain KC84, and strain KC19 was identified as belonging to the same genus as strain KC84, Bifidobacterium spp.

[0120] 16S rRNA (5'->3') SEQ ID NO: TGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTGTGTTGCCAGCACGTCGTGGTGGGAACTCACGGGG GACCGCCGGGGTCAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAACGGGATGCGACACTGTGAGGTGGAG CGGATCCCTTAAAACCGGTCTCAGTTCGGATTGGAGTCTGCAACCCGACTCCATGAAGGCGGAGTCGCTAGTAATCGCGGATCAGCAACGCCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGTAGCACCCGAAGCCGGTGGCCCAACCTTTTTGGGGGGAGCCGTCTAAGGTGAGACTCGTGATTGGGACTAAGTCGTAACAAGGTAACCGTAA1

[0121] Strains KC84, KC109, and KC19 were each cultured in BL medium under anaerobic conditions for 72 hours, and subsequently subcultured in the same medium under anaerobic conditions for 72 hours. The cultured microorganisms were centrifuged (12,000 xg, 1 min, 4°C), the supernatant was removed, and the cultures were resuspended. The resuspended microorganisms were 10 -4 Serial dilution was performed up to [value], and 10 μM Syto9 was treated and stained in a dark room for 15 minutes. Syto9 was analyzed using flow cytometry. +The number of microorganisms was measured by cell counting, and the number of strains was normalized using that value. To ensure washing and volume consistency, the microorganisms were centrifuged (10,000 xg, 10 min, 4°C), and after removing the supernatant, they were resuspended in PBS. To pasteurize the microorganisms, they were reacted in a 70°C water bath for 15 minutes, then centrifuged (10,000 xg, 10 min, 4°C), and after removing the supernatant, the remaining pellet was stored in a deep freezer (-80°C) until use in the experiment.

[0122]

[0123] Example 2. Serotonin secretion inhibitory effect

[0124] RIN14B (ATCC CRL-2059) cell lines were cultured in RPMI1640 medium supplemented with 10% FBS, penicillin (100 μg / mL), and streptomycin (100 μg / mL) at 37°C and 5% CO2, and subcultured every 3 days. RIN14B cell lines were placed in 96-well plates at a density of 7.4 x 10⁶ 4 Cells were dispensed per well and cultured for 48 hours.

[0125] The strain pellet prepared in Example 1 (1:100 relative to cells) or 20 μM of telotristat ethyl was suspended in RIN14B cell culture medium to prepare for treatment. After removing the medium from the RIN14B cell line dispensed into a 96-well plate, the cells were washed twice with 300 μL of HBSS + 2 μM fluoxetine per well. 100 μL of the prepared strain or telotristat ethyl was added, and the cells were incubated for 24 hours at 37°C under 5% CO2 conditions. The culture supernatant was collected, and filtration was performed using a filter plate and a vacuum manifold.

[0126] Serotonin secretion was measured using LC / MS-MS. Acetaminophen was added to the supernatant as an internal standard to a final concentration of 500 ng / mL and used as the measurement sample. A mixture of 0.1% formic acid in distilled water (mobile phase A) and acetonitrile (mobile phase B) was used as the mobile phase solution for liquid chromatography, and an HSS T3 column was used for the separation of serotonin. The separated analytes were detected using an Agilent 6546 qToF.

[0127] As shown in Figure 1, the group treated with the Bifidobacterium stuccoris KC84 strain according to one example of the present application showed about 92% serotonin secretion inhibitory ability compared to the untreated control group, and the serotonin secretion inhibitory effect was significantly superior even when compared to telotristat ethyl, which is known as a conventional serotonin inhibitor.

[0128] In addition, the Bifidobacterium stuccoris KC109 strain belongs to the same species as the Bifidobacterium stuccoris KC84 strain according to one example of the present application, but exhibited a serotonin secretion inhibitory ability of approximately 65%. Accordingly, the Bifidobacterium stuccoris KC84 strain according to one example of the present application showed a significantly superior serotonin secretion inhibitory effect even when compared to the Bifidobacterium stuccoris KC109 strain, which belongs to the same species. This means that the Bifidobacterium stuccoris KC84 strain according to one example of the present application has a significantly superior serotonin secretion inhibitory effect among Bifidobacterium stuccoris strains.

[0129] In addition, although the Bifidobacterium KC19 strain belongs to the same genus as the Bifidobacterium stuccoris KC84 strain according to one example of the present application, the serotonin secretion amount actually increased in the group treated with the Bifidobacterium KC19 strain. This means that the serotonin secretion inhibitory ability of the Bifidobacterium stuccoris KC84 strain according to one example of the present application is a strain-specific effect.

[0130] Accordingly, the excellent serotonin secretion inhibitory ability of Bifidobacterium stuccoris KC84 according to one example of the present application has been proven.

[0131]

[0132] Example 3. Inhibitory effect on mast cell degranulation

[0133] RBL-2H3 (ATCC CRL-2256) cell lines were cultured at 37°C under 5% CO2 conditions in DMEM medium supplemented with 10% FBS, penicillin (100 μg / mL), streptomycin (100 μg / mL), 10 mM HEPES, 1X NEAA, and 0.1% sodium bicarbonate, and subcultured every 2 days. RBL-2H3 cell lines were placed in 96-well plates at a rate of 2X10 5 Cells were seeded per well and cultured for 24 hours.

[0134] The strain pellet prepared in Example 1 (1:100 relative to cells) was suspended in siraganian buffer to prepare for treatment. After removing the medium from the RBL-2H3 cell line dispensed into a 96-well plate, the cells were washed twice with 200 μL of siraganian buffer per well. 90 μL of the prepared strain was added and incubated at 37°C for 20 minutes. Subsequently, the calcium ionophore A23187 was used as a degranulation stimulator. 10 μL of A23187 was added to achieve a final concentration of 5 μM and incubated at 37°C for 20 minutes. Afterward, the culture supernatant was collected and filtered using a filter plate and a vacuum manifold.

[0135] The ability to reduce degranulation was measured using a β-hexosaminidase assay. p-Nitrophenyl-N-acetyl-β-glucosaminide (PNAG) was prepared by dissolving it in citrate buffer (pH 4.5) to a concentration of 3.5 mg / mL (sonicated for 20 minutes). 50 μL each of the obtained culture supernatant and PNAG were added and incubated at 37°C for 2 hours. Subsequently, 50 μL of sodium biocarbonate (pH 10) was added, and the optical density (OD) was measured using a spectrophotometer. 405 ) was measured.

[0136] As shown in Fig. 2, the group treated with the Bifidobacterium stuccoris KC84 strain according to one example of the present application showed a reduction in mast cell degranulation of about 59% compared to the untreated control group.

[0137] In addition, the Bifidobacterium stuccoris KC109 strain belongs to the same species as the Bifidobacterium stuccoris KC84 strain according to one example of the present application, but showed an ability to reduce mast cell degranulation by about 35%. Accordingly, the Bifidobacterium stuccoris KC84 strain according to one example of the present application showed a significantly superior mast cell degranulation inhibitory effect even when compared to the Bifidobacterium stuccoris KC109 strain belonging to the same species. This means that the Bifidobacterium stuccoris KC84 strain according to one example of the present application shows a significantly superior mast cell degranulation inhibitory effect among Bifidobacterium stuccoris strains.

[0138] In addition, although the Bifidobacterium genus KC19 strain belongs to the same genus as the Bifidobacterium stuccoris KC84 strain according to one example of the present application, the mast cell degranulation rate of the group treated with the Bifidobacterium genus KC19 strain actually increased. This means that the mast cell degranulation inhibitory ability of the Bifidobacterium stuccoris KC84 strain according to one example of the present application is a strain-specific effect.

[0139] Accordingly, the excellent mast cell degranulation reduction ability of the Bifidobacterium stuccoris KC84 strain according to one example of the present application has been proven.

[0140]

[0141] Example 4. Preventive and therapeutic effects of irritable bowel syndrome (1)

[0142] (1) Manufacture of 5-HTP mouse model

[0143] The freeze-dried strain powder prepared in Example 1 was suspended in PBS with 0.05% cysteine ​​added, and then stirred for 30 minutes under anaerobic conditions at room temperature.

[0144] To prepare a 5-HTP mouse model, the KC84 strain according to one example of the present application was administered daily at a rate of 1 x 10 starting 7 days before inducing the disease. 9 It was administered orally at a CFU / head dose (Day 1–Day 7). Subsequently, for one week (Day 8–Day 14), 5-HTP dissolved in PBS was administered at 10 mg / kg (ip), and during the same period, the KC84 strain according to one example of the present application (1x10 9 CFU / head, po) or ondansetron (2 mg / kg, po), which is prescribed as a treatment for the relief of symptoms of diarrheal irritable bowel syndrome (IBS-d), was administered as a positive control.

[0145]

[0146] (2) Measurement of diarrhea score and defecation frequency

[0147] To measure diarrhea scores and bowel frequency, fecal pellet harvesting was performed for one hour starting immediately after intraperitoneal administration of 5-HTP on Day 11 and Day 14. Each mouse was placed individually in an empty cage, and fecal pellet harvesting was conducted for one hour. Bowel frequency data was obtained by counting the number of collected fecal pellets (Fig. 3a). The collected fecal pellet samples were visually evaluated on a scale of 0 to 3 based on the moisture and shape of each pellet (0: normal stool, 1: soft stool, 2: unformed stool, 3: watery stool), and the total diarrhea score was calculated by summing the scores of each pellet (Fig. 3b).

[0148] As shown in FIG. 3a, by administering the Bifidobacterium stuccoris KC84 strain according to one example of the present application, the defecation frequency, which is an indicator of diarrhea, was reduced by about 28% compared to the untreated control group and was reduced to a level equivalent to or greater than that of the positive control, ondansetron.

[0149] In addition, as shown in Fig. 3b, the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application reduced the diarrhea score, an indicator of the degree of diarrhea, by about 41% compared to the untreated control group and reduced to a level equivalent to that of the positive control, ondansetron.

[0150]

[0151] (3) Measurement of stool moisture content

[0152] Before starting fecal pellet collection, 1.75 mL microtubes were labeled and weighed. To minimize the effects of urine contamination and evaporation during collection, the fecal sample was collected immediately after generation using a 1 mL syringe needle, placed into a tube, and the lid was closed. After collection was completed, the 1.75 mL microtube containing the fecal pellet sample was weighed, and the wet weight was calculated by subtracting the weight of the empty 1.75 mL microtube. Subsequently, the 1.75 mL microtube containing the wet fecal pellet was placed in an oven with the lid open to remove moisture (60°C, overnight). The next day, the 1.75 mL microtube containing the dried fecal pellet was weighed, and the dry weight was calculated by subtracting the weight of the empty 1.75 mL microtube. The stool water content was calculated using the following formula (Fig. 3c):

[0153] Stool water content(%) = 100

[0154] As shown in Fig. 3c, the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application significantly reduced the stool water content, an indicator of diarrhea, by approximately 3% compared to the untreated control group. In particular, the stool water content was significantly reduced only by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, demonstrating a superior effect compared to the positive control, ondansetron.

[0155]

[0156] (4) Histological verification

[0157] On Day 14, mice were sacrificed 30 minutes after the completion of the fecal pellet harvest, and colon tissues were obtained, fixed in formalin, and embedded in paraffin blocks. 4 μm thick tissue sections were cut, stained with hematoxylin and eosin, and three sections were examined under a light microscope to evaluate the colitis index based on the observer's findings. The evaluation items and scoring criteria are as shown in Table 2, and the colitis index was evaluated from 0 to a maximum of 20 points (Fig. 3d).

[0158] ItemDescriptionScoreAbnormalities of mucosal architectureNone (Normal)0Minimal for focal, not exceeding laminar propria1Mild abnormality, cystic dilation / aberrant crypts2Moderate or multifocal abnormalities3Severe, entire crypt and epithelium lost4Extent of inflammationNone0Minimal for focal, scattered cells (<10%)1Mild (10-25%)2Moderate inflammatory cells extending into the Submucosa3Severe, transmural leukocytic infiltrate from mucosa to serosa4Erosion or ulcerationNo erosion, ulceration, or granulation tissue0Minimal or focal, not exceeding lamina propria1Unequivocal erosion2Moderate, ulceration3Severe ulceration or granulation tissue4Epithelial regenerationComplete regeneration or normal tissue0Almost complete regeneration1Regeneration with crypt depletion2Surface epithelium not intact3No tissue repair4Percent involvement1-25%126-50%251-75%376-100%4Severe colitisGrade semi-quantitatively from 0 to 20Total colitis index range: 0 to 20

[0159] As shown in FIG. 3d, by administering the Bifidobacterium stuccoris KC84 strain according to one example of the present application, the colitis index was significantly reduced by about 36% compared to the untreated control group and was reduced to a level equivalent to or greater than that of the positive control, ondansetron.

[0160]

[0161] Example 5. Serotonin reduction effect (1)

[0162] (1) Serotonin-reducing effect of luminal contents

[0163] Luminal content was obtained from the mouse sacrifice of Example 4, and serotonin levels were measured. 10 μL of ice-cold 80% methanol was added per 1 mg of sample, and homogenization was performed using a tissue lyser (30 Hz, 5 min, twice). After centrifugation (15,000 xg, 10 min, 4℃), the supernatant was obtained and impurities were removed using a 0.22 μm filter.

[0164] 10 mM ammonium formate in 40% ACN (mobile phase A) and 10 mM ammonium formate in 90% ACN (mobile phase B) were mixed and used as the mobile phase solution for liquid chromatography, and a BEH amide column was used for the separation of serotonin. The separated analytes were detected using Agilent 6546 qToF.

[0165] As shown in Fig. 4a, it can be confirmed that the serotonin level in the luminal content was reduced by approximately 21% compared to the untreated control group by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, and this effect was not observed in the positive control group administered ondansetron. Accordingly, the serotonin-reducing effect of the Bifidobacterium stuccoris KC84 strain according to one example of the present application was significantly superior to that of conventional serotonin-inhibiting drugs.

[0166]

[0167] (2) Reduced serotonin effect that has moved from serum to luminal content

[0168] Due to the characteristics of the 5-HTP animal model, serotonin present in the bloodstream can move to the intestines and be released into the lumen contents. Accordingly, the serotonin reduction effect was verified by calculating the ratio of serotonin that moved from the serum to the lumen contents.

[0169] Blood was obtained by heart blood collection during the sacrifice of the mouse in Example 4, and serum was obtained using SST. 900 μL of ice-cold 80% methanol was added per 100 μL of sample and stirred, then reacted at -20°C for 1 hour with the sample spin-down. After centrifugation (15,000 xg, 10 min, 4°C), the supernatant was obtained and impurities were removed using a 0.22 μm filter.

[0170] 10 mM ammonium formate in 40% ACN (mobile phase A) and 10 mM ammonium formate in 90% ACN (mobile phase B) were mixed and used as the mobile phase solution for liquid chromatography, and a BEH amide column was used for the separation of serotonin. The separated analytes were detected using Agilent 6546 qToF.

[0171] Figure 4b shows the ratio of the amount of serotonin in the luminal content measured in (1) of Example 5 to the amount of serotonin in the serum.

[0172] As shown in Fig. 4b, administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application reduced not only the serotonin level in the luminal content but also the proportion of serotonin that moved from serum to the lumen was significantly reduced. This effect was a reduction of approximately 23% in the proportion of serotonin that moved from plasma to the lumen compared to the untreated control group, and was not observed in the positive control group administered ondansetron, so the serotonin-reducing effect of the Bifidobacterium stuccoris KC84 strain according to one example of the present application was superior to that of the positive control group.

[0173]

[0174] (3) The effect of serotonin moving from the colon to the luminal content

[0175] The colon of the mouse sacrificed in Example 4 was obtained, and the serotonin level was measured. 20 μL of ice-cold 80% methanol was added per 1 mg of sample, and homogenization (30 Hz, 5 min, twice) was performed using a tissue lyser. After centrifugation (15,000 xg, 10 min, 4℃), the supernatant was obtained and impurities were removed using a 0.22 μm filter.

[0176] 10 mM ammonium formate in 40% ACN (mobile phase A) and 10 mM ammonium formate in 90% ACN (mobile phase B) were mixed and used as the mobile phase solution for liquid chromatography, and a BEH amide column was used for the separation of serotonin. The separated analytes were detected using Agilent 6546 qToF.

[0177] As shown in Fig. 4c, administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application reduced not only the serotonin level in the luminal content but also the proportion of serotonin that moved from the colon to the lumen was significantly reduced. This effect was a reduction of approximately 41% in the proportion of serotonin that moved from the colon to the lumen compared to the untreated control group, and was not observed in the positive control group administered ondansetron, so the serotonin-reducing effect of the Bifidobacterium stuccoris KC84 strain according to one example of the present application was superior to that of the positive control group.

[0178]

[0179] (4) Histological verification

[0180] The tissue section obtained in (4) of Example 4 was treated with an antibody against serotonin (1:5000) and stained, and a veterinary pathologist examined it under a microscope and calculated the serotonin-positive area using ImageJ software (Fig. 4d).

[0181] As shown in Fig. 4d, administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application significantly reduced the amount of serotonin involved in the pathogenesis of IBS-d by approximately 56% compared to the untreated control group. In particular, the Bifidobacterium stuccoris KC84 strain according to one example of the present application exhibited a superior serotonin inhibitory effect compared to the positive control, ondansetron.

[0182]

[0183] Example 6. Verification of changes in gene expression

[0184] RNA was extracted from the colon tissue obtained in Example 5 (3) according to the manufacturer's instructions for the QIAGEN Rneasy Plus Mini Kit, cDNA was obtained, and quantitative RT-PCR was performed to compare the expression levels of the Slc6a4, Tph1, IL1b, and Tjp1 genes with the housekeeping gene Gapdh.

[0185] As shown in FIG. 5a, the expression level of the Slc6a4 gene increased and the expression level of the Tph1 gene decreased upon administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application. The Slc6a4 gene is a serotonin transporter (SERT) gene that promotes serotonin reabsorption, and the Tph1 gene is a tryptophan hydroxylase gene that converts tryptophan into 5-HTP and is involved in serotonin synthesis. Therefore, the Bifidobacterium stuccoris KC84 strain according to one example of the present application has the effect of reducing serotonin levels by regulating the expression of the Slc6a4 and Tph1 genes, thereby increasing the reabsorption of serotonin and decreasing its biosynthesis. These results are consistent with the results of Examples 2 and 5, in which serotonin levels in luminal content were reduced by administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application.

[0186] In addition, as shown in Fig. 5b, the expression level of the IL-1β gene was reduced by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application. IL-1β is a pro-inflammatory cytokine, and the anti-inflammatory effect of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, specifically the effect of reducing intestinal inflammation, has been demonstrated. Considering that inflammation is also involved in the pathogenesis of IBS-d, these results are consistent with the results of Example 4, in which various IBS-d symptoms were alleviated by the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application.

[0187] In addition, as shown in FIG. 5c, the expression level of the Tjp1 gene increased upon administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application. The Tjp1 gene is a tight junction gene, and the Bifidobacterium stuccoris KC84 strain according to one example of the present application has the effect of strengthening tight junctions of colon mucosal epithelial cells and strengthening gut barrier function. These results are consistent with the results of Example 4, in which diarrhea symptoms were alleviated upon administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application.

[0188] Accordingly, it was verified that by administering the Bifidobacterium stuccoris KC84 strain according to one example of the present application, gene expression changes in a direction that reduces serotonin, inflammation-related gene expression decreases, and gut barrier function-related gene expression increases.

[0189]

[0190] Example 7. Preventive and therapeutic effects of irritable bowel syndrome (2)

[0191] (1) Zymosan mouse model manufacturing

[0192] The freeze-dried strain powder prepared in Example 1 was suspended in PBS with 0.05% cysteine ​​added, and then stirred for 30 minutes under anaerobic conditions at room temperature.

[0193] According to one example of the present application, the KC84 strain was administered to mice 5x10 times daily 9 It was administered orally at a CFU / head dose. The positive control group used sulfasalazine (30 mg / kg), a 5-ASA class drug prescribed for the relief of irritable bowel syndrome symptoms and the treatment of inflammatory bowel disease, and it was administered orally daily. For disease induction, zymosan (300 mg / kg) dissolved in PBS was administered via intracolonic injection on Day 1, Day 2, and Day 3.

[0194]

[0195] (2) Measurement of stool moisture content

[0196] On Day 4, Day 5, and Day 6, fecal pellet harvesting was carried out in substantially the same manner as in (3) of Example 4, and the stool water content was calculated and shown in FIG. 6.

[0197] As shown in Fig. 6, by administering the Bifidobacterium stuccoris KC84 strain according to one example of the present application, the stool moisture content, which is an indicator of the degree of diarrhea, was significantly reduced by about 7% compared to the untreated control group.

[0198]

[0199] Example 8. Effect of increasing intestinal butyrimonasabundance

[0200] Mice were sacrificed on Day 9 of Example 7, and cecums were obtained for amplicon sequencing. Microbiome analysis was performed on the fastaq files obtained through sequencing using the bioinformatic tool QIIME 2. The raw sequence data was demultiplexed and quality-filtered using the q2-demux plugin, and amplicon sequence variants (ASVs) were obtained using the DADA2 algorithm. All ASVs were aligned using the q2-alignment mafft, and phylogeny was obtained using the q2-phylogeny fasttree2. Taxonomic information regarding ASVs was classified using the q2-feature-classifier classify-sklearn naive Bayes taxonomy classifier and by referring to the Silva 13_8 99% OTUs reference sequences. Alpha-diversity (Shannon), beta-diversity (weighted UniFrac), and Principle Coordinate Analysis (PCoA) values ​​were estimated using q2-diversity after rarefying with 30,655 sequences per sample. LefSe analysis was performed using the open source server Galaxy (http: / galaxy.biobakery.org / ) with LDA 2.0 and a P-value of 0.05. The results of the α-diversity analysis are shown in Figure 7a, the results of the β-diversity analysis in Figure 7b, and the results of the LEfSe analysis in Figure 7c.

[0201] As shown in FIGS. 7a and 7b, α-diversity and β-diversity were not statistically significant upon administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application. On the other hand, as shown in FIG. 7c, the abundance of Butyrimonas increased upon administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application. Butyrimonas strains are known to produce butyrate, which lowers intestinal pH to inhibit the growth of pathogenic bacteria and contributes to maintaining beneficial microbial communities. Butyrate promotes the growth of intestinal epithelial cells, reduces the permeability of the intestinal barrier to prevent pathogens and harmful substances from entering the body, and has preventive and therapeutic effects on inflammatory bowel diseases through its anti-inflammatory action. Accordingly, the Bifidobacterium stuccoris KC84 strain according to one example of the present application has an effect on improving the intestinal microbiome.

[0202]

[0203] Example 9. Preventive and therapeutic effects of irritable bowel syndrome (3)

[0204] (1) WAS rat model manufacturing

[0205] The freeze-dried strain powder prepared in Example 1 was suspended in PBS with 0.05% cysteine ​​added, and then stirred for 30 minutes under anaerobic conditions at room temperature.

[0206] Before disease induction (D-2, D-1, D0), the KC84 strain according to one example of the present application was administered to rats at a rate of 1 x 10 times daily. 9It was administered orally at a CFU / head dose. Subsequently, the bacterial strain or alosetron (1.5 mg / kg), a serotonin type 3 (5-HT3) receptor selective antagonist prescribed to women for the relief of symptoms of diarrheal irritable bowel syndrome (IBS-d), was administered orally daily as a positive control, and water avoidance stress was applied for one hour. The number of fecal pellets produced during the one hour of stress was recorded. To measure the Diarrhea score, the bacterial strain and the positive control were administered on Day 6, Day 8, and Day 11, and fecal pellets were harvested for two hours.

[0207]

[0208] (2) Measurement of diarrhea score and defecation frequency

[0209] Using collected fecal pellet samples, each pellet was evaluated from 0 to 3 points in substantially the same manner as in (2) of Example 4, and the scores of each pellet were summed to calculate the total diarrhea score (Fig. 8a). The number of harvested pellets was counted and expressed as fecal pellet output of diarrhea score data (Fig. 8b). In addition, the number of feces produced during one hour of daily stress was counted and expressed as fecal pellet out data (Fig. 8c).

[0210] As shown in Fig. 8a, the administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application significantly reduced the diarrhea score, an indicator of the degree of diarrhea, by about 85% compared to the untreated control group, and this effect was significantly superior to that of the positive control, alosetron.

[0211] In addition, as shown in Fig. 8b, the FPO (fecal pellet output) during harvest was also significantly reduced by about 80% compared to the untreated control group by administering the Bifidobacterium stuccoris KC84 strain according to one example of the present application, and this effect was significantly superior to that of the positive control, alosetron.

[0212] In addition, as shown in Fig. 8c, the mean fecal pellet output was significantly reduced by about 44% compared to the untreated control group by administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, and this effect was significantly superior to that of the positive control, alosetron.

[0213]

[0214] Example 10. Serotonin reduction effect (2)

[0215] In Example 9, rats were sacrificed after the fecal pellet harvest on Day 11, and luminal content was obtained and used for serotonin measurement.

[0216] Ice-cold 80% methanol was added to the sample, and homogenization was performed using a tissue lyser (30 Hz, 5 min, twice). After centrifugation (15,000 xg, 10 min, 4℃), the supernatant was obtained and impurities were removed using a 0.22 μm filter.

[0217] 10 mM ammonium formate in 40% ACN (mobile phase A) and 10 mM ammonium formate in 90% ACN (mobile phase B) were mixed and used as the mobile phase solution for liquid chromatography, and a BEH amide column was used for the separation of serotonin. The separated analytes were detected using Agilent 6546 qToF.

[0218] As shown in Fig. 9, the serotonin level in the luminal content was significantly reduced by about 24% compared to the untreated control group by administration of the Bifidobacterium stuccoris KC84 strain according to one example of the present application, and was reduced to a level equivalent to or greater than that of the positive control, alosetron.

[0219]

[0220] Example 11. Antibiotic Resistance Test

[0221] According to one example of the present application, the Bifidobacterium stuccoris KC84 strain was cultured on MRS agar medium for 48 hours, and the strain was diluted to a concentration of 1.0 McFarland using PBS. The prepared bacterial cells were evenly spread onto LSM (LAB susceptibility test medium) agar plates (90% IST broth, 10% MRS broth, 1.8% agar) using a sterile cotton swab to form a lawn. The LSM agar plates inoculated with bacterial cells were left at room temperature for 10–15 minutes to allow the inoculated cells to permeate well. After placing the antibiotic strip to be tested on the LSM agar plates inoculated with bacterial cells, the plates were incubated at 37°C for 48 hours. For the determination of the MIC (minimal inhibitory concentration) in the E-test, the lowest concentration of the elliptical inhibition zone crossing the numerical scale of the strip was determined as the MIC. The results of the antibiotic resistance test are shown in Fig. 10.

[0222] As shown in FIG. 10, the antibiotic resistance (ampicillin;AM, vancomycin;VA, gentamycin;GM, streptomycin;SM, erythromycin;EM, clindamycin;CM, tetracycline;TC, chloramphenicol;CL) of the Bifidobacterium stuccoris KC84 strain according to one example of the present application was found to be below the EFSA standard, confirming that it is a safe strain.

[0223]

[0224] Example 12. Hemolytic activity test

[0225] A strain of Bifidobacterium stuccoris KC84 according to one example of the present application, cultured on MRS agar medium for 48 hours, was streaked onto blood agar medium and cultured under anaerobic conditions at 37°C for 24 to 48 hours. At this time, Staphylococcus aureus subsp. aureus ATCC12600 was used as a positive control after being cultured under aerobic conditions for 24 hours. Hemolytic activity was determined based on whether a clear halo formed around the colony. The observation results are shown in Fig. 11.

[0226] As shown in Fig. 11, no hemolytic activity was observed in the Bifidobacterium stuccoris KC84 strain according to one example of the present application, and its stability was confirmed.

[0227]

[0228] Example 13. Cytotoxicity Evaluation

[0229] According to one example of the present application, the Bifidobacterium stuccoris KC84 strain and the Staphylococcus aureus strain as a positive control were each cultured for 18 hours in MRS medium supplemented with 0.05% cysteine, and subsequently subcultured in the same medium for 18 hours. The cultured microorganisms were centrifuged (12,000 xg, 1 min, 4°C), the supernatant was removed, and the cultures were resuspended. The resuspended microorganisms were 10 -4 Serial dilation was performed, followed by treatment with 10 μM Syto9 and staining in a dark room for 15 minutes. Syto9 was analyzed using flow cytometry. +The number of microorganisms was measured by cell counting, and the number of strains was normalized using that value. To ensure washing and equal volume, the microorganisms were centrifuged (10,000 Xg, 10 min, 4℃), the supernatant was removed, and the culture was resuspended in MEM medium.

[0230] RIN14B (ATCC CRL-2059) cell lines were cultured in MEM medium supplemented with 20% FBS, penicillin (100 μg / mL), and streptomycin (100 μg / mL) at 37°C and 5% CO2, and subcultured every 3 days. RIN14B cell lines were placed in 96-well plates at a density of 1 x 10⁶ 4 Cells were dispensed per well and cultured for 24 hours.

[0231] The above-prepared Bifidobacterium stuccoris KC84 and Staphylococcus aureus strains were treated to Caco-2 cell lines inoculated into 96-well plates (at cell ratios of 1:10, 1:100, and 1:1000), respectively, and cultured for 24 hours at 37°C under 5% CO2 conditions. After collecting the culture supernatant, filtration was performed using a filter plate and vacuum manifold. The lysate control was obtained by adding 100 μL of RIPA lysis and extraction buffer to the cell lines, reacting for 30 minutes, and then filtering. The amount of NADH in the culture supernatant was measured according to the manual of the Lactate Dehydrogenase (LDH) Assay Kit, converted to a relative value based on 100% of the lysate control value, and is shown in Figure 12.

[0232] As shown in FIG. 12, the Bifidobacterium stuccoris KC84 strain according to one example of the present application did not exhibit cytotoxicity, and thus it was verified that it is a safe strain.

[0233] [Consignment Number]

[0234] Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resource Center (KCTC)

[0235] Trustee Number: KCTC15975BP

[0236] Date of Trust: 20240730

[0237]

Claims

1. Bifidobacterium stercoris strain KC84 having accession number KCTC15975BP.

2. In claim 1, the strain is a strain having the 16S rRNA sequence of SEQ ID NO.

1.

3. In claim 1, the strain has one or more features selected from the group consisting of (1) to (6) below: (1) Inhibits the secretion of serotonin; (2) Promotes the reabsorption of serotonin; (3) Inhibits the synthesis of serotonin; (4) Strengthening of tight junctions of colon mucosal epithelial cells; (5) Inhibition of degranulation of mast cells; and (6) Increase the relative abundance of Butyrimonas strains in the intestines.

4. A pharmaceutical composition for the prevention or treatment of serotonin syndrome, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

5. In paragraph 4, the composition is one that inhibits the secretion of serotonin of a subject.

6. In paragraph 4, the composition is one that promotes serotonin reabsorption of the subject.

7. In paragraph 4, the composition is one that inhibits serotonin synthesis of the subject.

8. In paragraph 4, the composition is one that reduces the serotonin concentration of the luminal content.

9. The composition according to claim 4, wherein the composition inhibits the movement of serotonin from the blood to the intestines.

10. The composition of claim 4, wherein the composition inhibits serotonin from moving from the colon to the lumen.

11. A pharmaceutical composition for the prevention or treatment of irritable bowel syndrome, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

12. A composition according to claim 11, wherein the irritable bowel syndrome is diarrhea-predominant irritable bowel syndrome.

13. The composition according to claim 11, wherein the composition reduces the moisture content of stool.

14. The composition of claim 11, wherein the composition reduces the frequency of bowel movements.

15. A pharmaceutical composition for the prevention or treatment of mast cell activation syndrome (MCAS), comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

16. A composition according to claim 15, wherein the mast cell activation syndrome is one or more selected from the group consisting of allergy, urticaria, rash, asthma, and chronic inflammation.

17. A composition for the prevention or treatment of inflammatory bowel disease comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

18. A composition for strengthening tight junctions of colon mucosal epithelial cells, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

19. A composition for increasing the relative abundance of Butyrimonas strains in the intestine, comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

20. A food comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having accession number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

21. A probiotic comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

22. A quasi-drug comprising one or more selected from the group consisting of a Bifidobacterium stercoris KC84 strain having deposit number KCTC15975BP, a culture of said strain, a lysate of said strain, and an extract of said strain.

Images