Intestinal microbiota improving agent containing barley processed product as an active ingredient

Abstract

The objective is to provide a gut microbiota-improving agent that can be taken on a daily basis. [Solution] An intestinal microbiota improving agent characterized by containing barley processed product as an active ingredient and adjusting the occupancy rate of one or more bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, the genus Anaerotruncus, the genus Mailhella, the family Muribaculaceae, the phylum Firmicutes, the genus Alicipipes, the family Lachnospiraceae, the genus Bacteroides, and the genus Blautia.

Description

Technical Field

[0001] The present invention relates to a gut microbiota improving agent containing a processed barley product as an active ingredient.

Background Art

[0002] In recent years, research on gut microbiota using metagenomic analysis technology has been actively conducted. One of the reasons is that the association between gut microbiota and diseases has been clarified. For example, the associations between gut microbiota and metabolic diseases such as obesity and diabetes, allergic diseases such as bronchial asthma and atopic dermatitis, cancer and the like have been reported.

[0003] The gut microbiota of animals such as humans is mainly composed of bacteria belonging to four phyla: Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria, and exists in various balances including other bacteria. However, when the diversity and balance of gut microbiota are lost due to changes in dietary habits such as a high-fat diet, it is thought to affect the intestinal mucosal immune system or induce inflammatory responses, thereby causing not only intestinal diseases but also systemic metabolic diseases and immune diseases.

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

Non-Patent Document 2

Direct Environment 7

Outdoor Track 8

Outdoor Tools9

Outdoor Tools 10

Outdoor Content11

Outdoor Tools 12

Non-Patent Document 16

Non-Patent Document 17

Summary of the Invention

[0005] The present invention aims to provide an intestinal microbiota improving agent that can be taken on a daily basis.

[0006] As a result of diligent research to solve the above problems, the inventors of this invention have discovered that by consuming barley processed products, diversity in the gut microbiota can be increased, and the proportion of bacteria involved in inflammatory diseases, lifestyle-related diseases, and improvement of quality of life can be regulated, thus completing the present invention. [Means for solving the problem]

[0007] The present invention includes the following embodiments. [1] An intestinal microbiota improving agent that contains barley processed product as an active ingredient and is characterized by adjusting the occupancy rate of one or more bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, the genus Anaerotruncus, the genus Mailhella, the family Muribaculaceae, the phylum Firmicutes, the genus Alistipes, the family Lachnospiraceae, the genus Bacteroides, and the genus Blautia. [2] This is an intestinal microbiota improving agent as described in [1], which reduces one or more bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, bacteria belonging to the genus Anaerotruncus, and bacteria belonging to the genus Mailhella. [3] This is an intestinal microbiota improving agent described in [1], which promotes the growth of one or more bacteria selected from the group consisting of bacteria belonging to the family Muribaculaceae, the family Lachnospiraceae, the phylum Firmicutes, the genus Bacteroides, the genus Alicipipes, and the genus Blautia. [4] Food and beverages, or intestinal microbiota improving agents as described in any of [1] to [3]. [5] The intestinal microbiota improving agent according to any one of [1] to [3], which is a pharmaceutical product. [6] The intestinal microbiota improving agent according to any one of [1] to [3], which is a pet food. [7] A preventive and / or ameliorating agent for inflammatory diseases, comprising a processed barley product as an active ingredient. [8] A preventive and / or ameliorating agent for lifestyle-related diseases, comprising a processed barley product as an active ingredient. [9] A QOL improving agent comprising a processed barley product as an active ingredient.

[10] The QOL improving agent according to [9], wherein QOL is one or more selected from the group consisting of physical health, mental health, and social health.

[11] A mucin production promoter comprising a processed barley product as an active ingredient.

[12] The mucin production promoter according to

[11] , which is for preventing and ameliorating diseases caused by decreased mucin.

[13] The mucin production promoter according to

[12] , wherein the disease is one or more selected from the group consisting of viral infections, bacterial infections, respiratory diseases, inflammatory bowel diseases, and dry eye. [Effects of the Invention]

[0008] The intestinal microbiota improving agent of the present invention improves the intestinal microbiota, enables prevention and amelioration of diseases such as inflammatory diseases and lifestyle-related diseases, and also enables improvement of QOL. As another aspect, the mucin production promoter of the present invention promotes mucin production, thereby enabling prevention and amelioration of diseases such as viral infections and bacterial infections caused by decreased mucin. [Brief Description of the Drawings]

[0009] [Figure 1]Figure 1 is a graph showing the results of alpha diversity analysis by Qiime2 for the intestinal microbiota of each group in Example 1. Figure 1(a) shows the Chao1 index, and Figure 1(b) shows the Shannon index. For statistical analysis, the Kruskal-Wallis test was followed by Dunn's test for multiple comparisons. "*" indicates a significant difference (*: p<0.05). [Figure 2] Figure 2 is a graph showing the results of beta diversity (Bray-Curtis) analysis by Qiime2 for the intestinal microbiota of each group in Example 1. [Figure 3] Figure 3 is a graph showing the results of alpha diversity analysis by EzBioCloud for the intestinal microbiota of each group in Example 1 (Chao1 index). [Figure 4] Figure 4 is a graph showing the results of alpha diversity analysis by EzBioCloud for the intestinal microbiota of each group in Example 1 (Shannon index). [Figure 5] Figure 5 is a graph showing the results of beta diversity analysis by EzBioCloud for the intestinal microbiota of each group in Example 1. [Figure 6] Figures 6(a) to (b) show the relative abundance of bacteria in each group in Example 1. "*" and "**" indicate significant differences (*: p<0.05, **: p<0.01). [Figure 7] Figures 7(a) to (e) show the relative abundance of bacteria in each group in Example 1. "*", "**", and "***" indicate significant differences (*: p<0.05, **: p<0.01, ***: p<0.001). [Figure 8] Figure 8 shows the mucin content contained in the feces of each group in Example 2. [Figure 9] Figures 9(a) to (e) show the results of a questionnaire conducted to evaluate the QOL-improving effect of barley powder intake. "*" indicates a significant difference (*: p<0.05). [Figure 10] Figure 10 shows the relative abundance of Blautia_A_141781 before and after barley powder intake. "*" indicates a significant difference (*: p<0.05). [Modes for carrying out the invention]

[0010] The present invention will be described in detail below. The description of the present invention below may be based on preferred embodiments of the present invention, but the present invention is not limited to such embodiments. In this specification, the "~" symbol indicating a numerical range is used to mean that the numbers before and after it are included as the lower and upper limits, respectively.

[0011] The intestinal microbiota improving agent of the present invention contains barley processed product as an active ingredient.

[0012] Barley (Hordeum vulgare), used in barley products, is an annual or super-annual herbaceous grain belonging to the grass family (Poaceae), native to Central Asia. Barley is classified into "hulled barley" and "naked barley" based on how easily the outer husk of the grain is removed, and each of these is further classified into "two-row barley" and "six-row barley." Two-row barley is barley in which the spikelets are arranged in two rows around the axis of the ear. Specific varieties include two-row nalled barleys such as Waxy Fiber, View Fiber, Kirarimochi, and Yumesakiboshi; and two-row hulled barleys such as Haruka Nijo, Shirotae Nijo, Koharu Nijo, Kira Nijo, Kirinijo, and Nishinohoshi. "Six-row barley" refers to barley in which spikelets are arranged in six rows around the axis of the ear. Specific varieties include, for example, six-row hulless barley such as Daikinboshi, Haruakane, Fukumi Fiber, Nagasaki Mishima, and Haruhimeboshi; and six-row hulled barley such as Yukihana Rokujo, Yukimi Rokujo, and Kashima Gold. In the present invention, any of the above classifications and varieties of barley may be used for the barley processed product, but for example, the improved variety barley BarleyMax® is preferably used.

[0013] There are no particular restrictions on the processed barley product; the grain may be used as is, or crushed or pulverized grains may be used, obtained by crushing or pulverizing the grain using known methods. The grain may be either whole grain barley with the outer husk intact, or polished barley with the outer husk removed.

[0014] The intestinal microbiota improving agent of the present invention has the effect of reducing one or more bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, bacteria belonging to the genus Anaerotruncus, and bacteria belonging to the genus Mailhella in the intestinal microbiota.

[0015] Bacteria belonging to the phylum Desulfobacterota (hereinafter referred to as "Desulfobacterota bacteria") are a bacterial phylum that includes many sulfate-reducing bacteria (SRBs), which reduce sulfates and produce hydrogen sulfide, etc. Families belonging to the phylum Desulfobacterota include, for example, the families Desulfobacteraceae, Desulfovibrio, Desulfomicrobiaceae, Desulfobulbaceae, and Desulfuromonadaceae. Each family includes bacteria such as the genera Desulfobacter in the family Desulfobacteraceae, Desulfovibrio in the family Desulfovibrio, Desulfomicrobium in the family Desulfomicrobiaceae, Desulfobulbus in the family Desulfobulbaceae, and Desulfuromona in the family Desulfuromonadaceae.

[0016] Regarding Desulfobacterota bacteria, for example, it has been reported that patients with inflammatory bowel disease and irritable bowel syndrome have a higher proportion of Desulfovibrio bacteria compared to healthy individuals (Non-Patent Literature 1). The intestinal microbiota improving agent of the present invention can reduce the proportion of Desulfobacterota bacteria in the intestinal microbiota and prevent and / or improve intestinal diseases such as inflammatory bowel disease and irritable bowel syndrome.

[0017] Bacteria belonging to the genus Anaerotruncus (hereinafter referred to as "Anaerotruncus bacteria") are Gram-positive anaerobic bacteria belonging to the family Oscillospiraceae. It has been reported that Anaerotruncus bacteria occupy a higher proportion in frail elderly individuals compared with non-frail elderly individuals (Non-Patent Literature 2). It has also been reported that Anaerotruncus bacteria occupy a higher proportion in patients with chronic renal failure compared with healthy individuals (Non-Patent Literature 3). The intestinal microbiota improving agent of the present invention can reduce the proportion of Anaerotruncus bacteria in the intestinal microbiota and prevent and / or improve frailty and renal failure.

[0018] Bacteria belonging to the genus Mailhella (hereinafter referred to as "Mailhella bacteria") are Gram-positive anaerobic bacteria belonging to the family Desulfovibrionaceae, and examples include Mailhella massiliensi, Candidatus, and Mailhella excrementigallinarum. Mailhella bacteria have been reported to be useful as predictors of hyperplatelet reactivity (HTPR) during treatment in patients with acute ischemic stroke (AIS) (Non-Patent Literature 4). The gut microbiota improving agent of the present invention can prevent hyperplatelet reactivity and further prevent recurrence of acute ischemic stroke by reducing the proportion of Mailhella bacteria in the gut microbiota.

[0019] The intestinal microbiota improving agent of the present invention has the effect of increasing one or more bacteria selected from the group consisting of bacteria belonging to the family Muribaculaceae, bacteria belonging to the family Lachnospiraceae, bacteria belonging to the phylum Firmicutes, bacteria belonging to the genus Bacteroides, bacteria belonging to the genus Alicipipes, and bacteria belonging to the genus Blautia.

[0020] Regarding bacteria belonging to the family Muribaculaceae (hereinafter referred to as "Muribaculaceae bacteria"), it has been reported that in mice with dextran sulfate sodium (DSS)-induced colitis, the proportion of Muribaculaceae bacteria is lower compared to the control group (Non-Patent Literature 5). Furthermore, inflammatory bowel diseases such as Crohn's disease and ulcerative colitis develop due to the excessive production and induction of inflammatory cytokines such as TNF-α, IL-6, and IL-1β, and studies using DSS-induced colitis mice have shown a negative correlation between the proportion of Muribaculaceae bacteria and the amount of inflammatory cytokines. The gut microbiota improving agent of the present invention can increase the proportion of Muribaculaceae bacteria in the gut microbiota and prevent and / or improve inflammatory bowel diseases such as Crohn's disease and ulcerative colitis induced by inflammatory cytokine production.

[0021] Furthermore, it has been reported that type 2 diabetic mice have a lower proportion of Muribaculaceae bacteria compared to healthy mice (Non-Patent Literature 5). In addition, a negative correlation has been reported between the risk of obesity and Muribaculaceae bacteria (Non-Patent Literature 5). The gut microbiota improving agent of the present invention can increase the proportion of Muribaculaceae bacteria in the gut microbiota and prevent and / or improve type 2 diabetes and obesity.

[0022] The Muribaculaceae family includes, for example, bacteria of the genera Muribaculum, Duncanella, Paramuribaculum, and Metamuribaculum. In particular, the intestinal microbiota improving agent of the present invention can suitably increase UBA7173_g, Paramuribaculum bacteria, PAC001066_g, PAC001068_g, PAC001112_g, and PAC002448_g in the intestinal microbiota and increase their proportion. Here, "UBA7173_g" refers to the bacteria listed in the database "GreenGene2 (Ver.2024.09)". Also, "PAC001066_g", "PAC001068_g", "PAC001112_g", and "PAC002448_g" refer to the bacteria listed in "EZBioCloud Database (Ver.2023.08.23)".

[0023] Regarding bacteria belonging to the family Lachnospiraceae (hereinafter referred to as "Lachnospiraceae bacteria"), it has been reported that the proportion of Lachnospiraceae bacteria is lower in patients with Crohn's disease and ulcerative colitis compared to healthy individuals (Non-patent documents 6 and 7). The intestinal microbiota improving agent of the present invention can increase the proportion of Lachnospiraceae bacteria in the intestinal microbiota and prevent and / or improve inflammatory bowel diseases such as Crohn's disease and ulcerative colitis.

[0024] The Lachnospiraceae family includes, for example, bacteria of the genera Roseburia, Blautia, Lachnospira, Anaerostipes, Dorea, Coprococcus, and Fusimonas. In particular, the intestinal microbiota improving agent of the present invention can suitably increase Blautia_A_141781, Fusimonas bacteria, COE1_g, Eubacterium_g17, and PAC001165_g, and increase their proportion. Here, "Blautia_A_141781" refers to the bacteria listed in the database "GreenGene2 (Ver.2025.04)". "COE1_g" refers to the bacteria listed in the database "GreenGene2 (Ver.2024.09)". Furthermore, "Eubacterium_g17" and "PAC001165_g" refer to bacteria listed in the "EZBioCloud Database (Ver.2023.08.23)".

[0025] Bacteria belonging to the genus Blautia (hereinafter referred to as "Blautia bacteria") are Gram-positive anaerobic bacteria and have been reported to contribute to the improvement of quality of life (QOL) (Non-Patent Literature 8). In this specification, "improvement of quality of life" means improvement of at least one health condition selected from the group consisting of physical health, mental health, and social health. Specifically, improvement of physical health means reduction of pain, fatigue, and malaise, improvement of sleep quality, increased vitality, and improvement of bowel movements (e.g., frequency of bowel movements, feeling of incomplete evacuation, odor); improvement of mental health means reduction of depression, reduction of irritability, improved concentration, and increased self-esteem; and improvement of social health means improvement of interpersonal relationships and increased willingness to participate in social activities. The intestinal microbiota improving agent of the present invention can increase the occupancy rate of Blautia bacteria in the intestinal microbiota and improve QOL.

[0026] Bacteria belonging to the phylum Firmicutes (hereinafter referred to as "Firmicutes bacteria") are one of the major bacteria constituting the gut microbiota in mammals such as humans and mice, and are characterized by low levels of guanine (G) and cytosine (C) in their DNA. Examples of classes belonging to the phylum Firmicutes include Clostridia, Bacilli, Erysipelotrichia, and Nagativecutes. It has been reported that the proportion of Firmicutes bacteria is lower in Crohn's disease patients compared to healthy individuals (Non-Patent Literature 8). The gut microbiota improving agent of the present invention can increase the proportion of Firmicutes bacteria in the gut microbiota and prevent and / or improve inflammatory bowel diseases such as Crohn's disease.

[0027] Bacteria belonging to the genus Bacteroides (hereinafter referred to as "Bacteroides bacteria") are Gram-negative anaerobic bacteria belonging to the family Bacteroidaceae. It has been reported that the proportion of Bacteroides bacteria is lower in patients with inflammatory bowel disease compared to healthy individuals (Non-Patent Literature 9). It has also been reported that the proportion of Bacteroides bacteria is lower in obese individuals compared to lean individuals (Non-Patent Literature 10). The intestinal microbiota improving agent of the present invention can increase the proportion of Bacteroides bacteria in the intestinal microbiota and prevent and / or improve inflammatory bowel disease and obesity.

[0028] Examples of bacteria of the genus Bacteroides include Bacteroides fragilis, Bacteroides acidifaciens, Bacteroides thetaiotaomicron, and Bacteroides vulgatus. The intestinal microbiota improving agent of the present invention can suitably increase Bacteroides acidifaciens.

[0029] Bacteria belonging to the genus Alistipes (hereinafter referred to as "Alistipes bacteria") are Gram-negative anaerobic bacteria belonging to the family Rikenellaceae. It has been reported that the proportion of Alistipes bacteria is lower in colorectal cancer patients compared to healthy individuals (Non-Patent Literature 11). It has also been reported that insulin resistance, one of the factors that contribute to the development of type 2 diabetes, can be improved by increasing the proportion of Alistipes bacteria (Non-Patent Literature 12). The intestinal microbiota improving agent of the present invention can increase the proportion of Alistipes bacteria in the intestinal microbiota and prevent and / or improve colorectal cancer and type 2 diabetes.

[0030] The intestinal microbiota improving agent of the present invention uses barley processed product as an active ingredient. The content of barley processed product in the intestinal microbiota improving agent of the present invention is not particularly limited, but for example, it is 1 to 99.9% by amount, preferably 1 to 99% by mass, and more preferably 1 to 98% by mass.

[0031] The intestinal microbiota improving agent of the present invention can be formulated by combining the above-mentioned active ingredient with a pharmaceutically acceptable carrier as needed. Examples of pharmaceutically acceptable carriers include glucose, lactose, starch, mannitol, dextrin, fatty acid glycerides, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid esters, amino acids, gelatin, albumin, water, physiological saline, etc. Furthermore, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, isotonic agents, and excipients may be added as needed. The dosage form is not particularly limited, but examples include liquids, powders, granules, capsules, tablets, etc., which can be manufactured according to conventional methods.

[0032] The intestinal microbiota improving agent of the present invention can be applied to mammals including humans, such as humans, dogs, cats, cattle, pigs, chickens, sheep, and horses, and is preferably used in humans. The dosage is not particularly limited, but for example, the daily dose for an adult (weighing 60 kg) is about 1 to 20 g, preferably 3 to 17 g, more preferably 5 to 15 g, and even more preferably 7 to 13 g. The number of doses may be, for example, once a day, or divided into multiple doses. The interval between doses may be, for example, daily, or every few days.

[0033] The intestinal bacteria improving agent of the present invention may also be in the form of food or beverages. It is prepared by combining the above-mentioned active ingredient with known food additives and / or food materials, according to conventional methods, and its form is not particularly limited. Examples include starch-based foods such as bread, biscuits, pancakes, noodles, and candy tablets; confectionery such as gum, candy, and Japanese sweets; meat products such as ham and sausages; fish products such as chikuwa and kamaboko; seafood products; seasonings such as dressings, soy sauce, jam, and furikake; and beverages such as tea, juice, soft drinks, and alcoholic beverages. Food and beverages include Foods for Specified Health Uses and Foods with Function Claims that have been permitted or notified for their use in improving the intestinal microbiota.

[0034] The intestinal microbiota improving agent of the present invention can also be in the form of feed or pet food, and is prepared by appropriately blending the above-mentioned active ingredient with known feed materials, gelling agents, shape-retaining agents, pH adjusters, seasonings, preservatives, nutritional supplements, etc., according to conventional methods. Its form is not particularly limited, and any form such as pellets, flakes, or mash are exemplified, and it can be used, for example, as feed for livestock such as cattle, pigs, chickens, sheep, and horses, or as pet food for dogs, cats, etc.

[0035] The intestinal microbiota improving agent of the present invention enhances the diversity of the intestinal microbiota and exerts preventive and ameliorative effects against inflammatory bowel diseases such as Crohn's disease, ulcerative colitis, and irritable bowel syndrome, as well as inflammatory diseases such as psoriasis, rheumatoid arthritis, nephritis, pneumonia, and multiple sclerosis; and lifestyle-related diseases such as obesity, diabetes, cerebrovascular disease, heart disease, dyslipidemia, and hypertension, by increasing the diversity of the intestinal microbiota and reducing bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, the genus Anaerotruncus, and the genus Mailhella, or by increasing bacteria selected from the group consisting of bacteria belonging to the families Muribaculaceae, Lachnospiraceae, Firmicutes, the phylum Bacteroides, the genus Bacteroides, and the genus Alistopes.

[0036] In another embodiment, the present invention is a mucin production promoter containing a barley product as an active ingredient. In the present invention, descriptions of components similar to those of the above-mentioned intestinal microbiota improving agent will be omitted as appropriate.

[0037] Mucin is a high-molecular-weight glycoprotein and is the main component of mucus produced by goblet cells in mucosal epithelium of the eyes, nasal cavity, digestive tract, and respiratory tract, as well as by salivary glands in the oral cavity. There are two types of mucin: secretory and transmembrane. Secretory mucin forms a mucus layer on the mucosal surface, inhibiting the entry of pathogens such as bacteria and viruses into the body. Transmembrane mucin is localized to the cell membrane and, like secretory mucin, inhibits the entry of pathogens into the body, as well as being involved in intercellular signal transduction. The mucin production promoter of the present invention can promote mucin production, enabling the prevention and improvement of bacterial and viral infections, as well as promoting intercellular signal transduction.

[0038] It has been reported that a decrease in mucin in the eye impairs the stability of the tear film and is involved in the development of dry eye and other conditions (Non-Patent Literature 14). It has been reported that a decrease in mucin in the nasal cavity is involved in the development of respiratory diseases such as allergic rhinitis (Non-Patent Literature 15). It has been reported that a decrease in mucin in the airways is involved in the development of respiratory diseases such as asthma and bronchitis (Non-Patent Literature 16). It has been reported that a decrease in mucin in digestive organs such as the stomach and intestines is involved in the development of inflammatory bowel diseases such as ulcerative colitis and Crohn's disease (Non-Patent Literature 17). The mucin production promoter of the present invention can prevent and improve these diseases by promoting mucin production. [Examples]

[0039] The present invention will be described in detail below with reference to examples. However, the present invention is not limited to these examples.

[0040] <Example 1: Effects of consuming barley processed products on the gut microbiota> To evaluate the effects of barley processed food intake on the gut microbiota, an analysis of the gut microbiota was performed. In this example, barley bucks processed into powder was used as the barley processed food (hereinafter referred to as "barley powder").

[0041] [Materials and Rearing Conditions] Six-week-old C57BL / 6 mice were purchased from SCL Japan Co., Ltd. and reared under conditions of 24°C ± 2°C, a 12-hour light-dark cycle, and free access to water and feed (AIN-93G; CREA Japan Co., Ltd.). Each cage was arranged to minimize environmental influence on the mice.

[0042] [Dosage Schedule] Under the above conditions, the mice underwent a one-week preliminary rearing period. At seven weeks of age, they were randomly divided into a control group (Ctrl group; 6 mice) and barley powder-treated groups (Low treatment group; 6 mice; Mid treatment group; High treatment group; 6 mice). Each group was fed the appropriate diet for four weeks. The control group was fed feed (AIN-93G; manufactured by CREA Nippon Co., Ltd.). The Low treatment group was fed feed containing 1.25% barley powder (BARLEYMax®, Teijin Limited), the Mid treatment group was fed feed containing 2.5% barley powder, and the High treatment group was fed feed containing 5% barley powder. The crude protein, crude fat, and soluble non-nitrogenous substances in the feed given to each group were adjusted to be equal, thereby ensuring that the diet energy of each group was also equal.

[0043] After the administration period ended, the mice from each group were dissected and their feces were collected.

[0044] (Intestinal microbiota analysis) For the analysis of the gut microbiota, DNA was extracted from feces collected using the following method. Frozen feces were thawed on ice, and 100 mg of each sample was suspended in a solution containing 4 M guanidia thiocyanate, 100 mM Tris-HCl (pH 9.0), and 40 mM EDTA. The suspensions were disrupted with zirconia beads using FastPrep FP100A (MP Biomedicals). DNA was extracted from the bead-treated suspensions using Magtration System 12GC and GC Series MagDEA DNA 200.

[0045] The obtained DNA was used as a template to amplify the 16S rRNA V3-V4 region using primers (Pro341F and Pro805R) via PCR. The resulting PCR products were analyzed using the paired-end method with the Illumina MiSeq platform and MiSeq Reagent Kit v3 (both from Illumina). The resulting nucleotide sequences were taxonomically assigned using the analysis platform "Qiime2" with GreenGenes2 as the reference database. For each group of gut microbiota, α-diversity analysis (Figure 1), β-diversity analysis (Figure 2), and calculation of the relative abundance of bacteria (Figures 6 and 7) were performed.

[0046] Furthermore, the nucleotide sequences obtained from the above analysis were taxonomically assigned using another analysis platform, "EzBioCloud (https: / / www.ezbiocloud.net / )," referencing the database (Ver.20230823). Then, α-diversity analysis (Figures 3 and 4), β-diversity analysis (Figure 5), and intergroup comparison analysis (Table 1) were performed on the gut microbiota of each group.

[0047] (α-diversity analysis) Alpha diversity analysis allows for the evaluation of the diversity of the gut microbiota within each group. In this example, the Chao1 Index and the Shannon Index were used as indicators. A higher index indicates greater diversity of the gut microbiota. As the dose of barley powder increased, it was confirmed that species diversity tended to increase compared to the control group (Figures 1, 3, and 4).

[0048] (β diversity analysis) β-diversity analysis allows us to evaluate the differences in gut microbiota diversity among the groups. The further apart the plots are, the more different the composition of the gut microbiota. A significant distance was observed between the plots of the control group and the barley powder administration group (Figures 2 and 5).

[0049] (Relative abundance of bacteria) Compared to the control group, a decrease in the relative abundance of Desulfobacterota bacteria was observed (Figure 6(a)). Furthermore, an increase in the relative abundance of Firmicutes bacteria was observed (Figure 6(b)).

[0050] As shown in Figure 7, compared to before barley powder intake, an increasing trend in the relative abundance of UBA7173_g (Muribaculaceae bacteria), Paramuribaculum bacteria (Muribaculaceae bacteria), and Lachnospriraceae COE1_g (Lachnospiraceae bacteria) was observed after intake (Figure 7(a)~(c)). Furthermore, a decrease in the relative abundance of Anaerotruncus and Mailhella bacteria was observed after ingestion compared to before ingestion (Figure 7(d), (e)).

[0051] (Group comparison analysis (LEfSe analysis)) We identified characteristic bacterial species in each group using intergroup comparative analysis (average relative abundance > 1%).

[0052] [Table 1]

[0053] As shown in Table 1, compared to before barley powder intake, an increase in the relative abundance of bacteria belonging to the families Muribaculaceae, Alistipes, Bacterioides acidifacies, and Lachnospiraceae was observed after intake.

[0054] <Example 2: Effect of barley processing on mucin content> To investigate the effect of barley processing on the mucin content in feces, mucin was quantified. In this example, as in Example 1, powdered BarleyMax was used as the barley processing material.

[0055] The materials, rearing conditions, and administration schedule in this embodiment are the same as in Example 1, so a detailed description is omitted.

[0056] (Measurement of mucin content) A fecal mucin measurement kit (manufactured by Cosmo Bio Co., Ltd.) was used to measure the mucin content in the feces of each group. The measurements were performed according to the kit's protocol. The results are shown in Figure 8.

[0057] As shown in Figure 8, it was confirmed that the amount of mucin in the feces increased after ingesting barley processed products compared to before ingesting them.

[0058] <Example 3: Effects of barley processed food intake on quality of life and gut microbiota> To evaluate the effects of barley processed food intake on quality of life (QOL) and gut microbiota, questionnaires and gut microbiota analysis were conducted. In this example, as in Example 1 above, BarleyMax processed into powder was used as the barley processed food.

[0059] [subject] This study involved 16 healthy individuals as subjects. The average age of the subjects was 21.9 years. Details of the subject selection and exclusion criteria are as follows.

[0060] [Selection criteria] 1. Men and women aged 18 to under 75 at the time of obtaining consent. 2. Individuals who do not have an intestinal disease (including those who are deemed not to require immediate treatment) 3. Individuals who have received a thorough explanation of this examination, understand its contents, and can give their consent.

[0061] [Exclusion criteria] 1. Individuals who may experience abdominal pain from dietary fiber-containing materials such as BarleyMax. 2. Individuals diagnosed with a digestive system disease, those who have undergone digestive system surgery in the past, or those who are expected to require digestive system surgery in the future. 3. Individuals who regularly consume dietary fiber, including BarleyMax (3 days or more per week), or who have regularly consumed it within the 3 months prior to the pre-examination. 4. Individuals diagnosed with malignant tumors, those currently being treated for neurological, heart, kidney, diabetes, or thyroid dysfunction, or those with a history of serious illnesses such as liver disease (excluding those whom the physician in charge determines will not affect the evaluation of this study). 5. Individuals who consume large amounts of alcohol (60g or more of alcohol per day: approximately 1500mL of beer, 540mL of wine, 3 go of sake, or 180mL of a double shot of whiskey). 6. Persons with allergies to wheat or barley, or other persons who may develop an allergic reaction to the research food. 7. Individuals deemed unsuitable as subjects based on their responses to the lifestyle questionnaire. 8. Individuals who have participated in other clinical research within one month prior to obtaining consent to participate in this study, or who are scheduled to participate in other clinical research after obtaining consent to participate in this study. 9. Other persons whom the person in charge of the program deems unsuitable as subjects.

[0062] [Intervention details] The subjects were given 4 grams of barley powder per day for 4 weeks.

[0063] (Subjective evaluation index) To evaluate the effect of barley powder intake on improving quality of life, a questionnaire was administered based on the following evaluation criteria for the following evaluation items (a) to (e). The results are shown in Figure 9.

[0064] (Evaluation criteria) (a) Do you have the energy to go about your daily life? (b) Are you satisfied with your health? (c) Are you satisfied with your sleep? (d) How healthy is your living environment? (e) Are you satisfied with your relationships?

[0065] (Evaluation Criteria) 0: None at all 1: There was a little 2: There were quite a few. 3: There were quite a few. 4: There were a lot

[0066] As shown in Figures 9(a) to (e), it was confirmed that the scores for each evaluation item improved after consuming barley powder compared to before consumption. This suggests that continuous consumption of barley powder has a positive effect on improving quality of life (QOL).

[0067] (Intestinal microbiota analysis) The analysis of the gut microbiota in this example was performed in the same manner as in Example 1, except that the reference database used was "2024.09.backbone.full-length.nb.sklearn-1.4.2.qza". The results are shown in Figure 10.

[0068] As shown in Figure 10, the relative abundance of Blautia_A_141781 was confirmed to have significantly increased after barley powder intake compared to before intake.

Claims

1. An intestinal microbiota improving agent characterized by containing barley processed product as an active ingredient and adjusting the occupancy rate of one or more bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, the genus Anaerotruncus, the genus Mailhella, the family Muribaculaceae, the phylum Firmicutes, the genus Alistipes, the family Lachnospiraceae, the genus Bacteroides, and the genus Blautia.

2. An intestinal microbiota improving agent according to claim 1, which reduces one or more bacteria selected from the group consisting of bacteria belonging to the phylum Desulfobacterota, bacteria belonging to the genus Anaerotruncus, and bacteria belonging to the genus Mailhella.

3. An intestinal microbiota improving agent according to claim 1, which increases one or more bacteria selected from the group consisting of bacteria belonging to the family Muribaculaceae, bacteria belonging to the family Lachnospiraceae, bacteria belonging to the phylum Firmicutes, bacteria belonging to the genus Bacteroides, bacteria belonging to the genus Alicipipes, and bacteria belonging to the genus Blautia.

4. An intestinal microbiota improving agent according to claim 1 or 2, which is a food or beverage.

5. A pharmaceutical product, an intestinal microbiota improving agent according to claim 1 or 2.

6. An intestinal microbiota improving agent according to claim 1 or 2, which is a pet food.

7. An agent for the prevention and / or improvement of inflammatory diseases, containing barley processed products as an active ingredient.

8. An agent for the prevention and / or improvement of lifestyle-related diseases, containing barley processed products as an active ingredient.

9. A quality of life (QOL) improving agent containing barley processed products as an active ingredient.

10. The QOL improving agent according to claim 9, wherein QOL is one or more selected from the group consisting of physical health, mental health, and social health.

11. A mucin production promoter containing barley processed products as an active ingredient.

12. A mucin production promoter according to claim 11, for preventing or improving diseases caused by a decrease in mucin.

13. The mucin production promoter according to claim 12, wherein the disease is one or more selected from the group consisting of viral infections, bacterial infections, respiratory diseases, inflammatory bowel diseases, and dry eye.

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