JP6247185B2 - Vylophila bacterium growth inhibitor and method for suppressing virulence bacteria growth - Google Patents

Description

  The present technology relates to an agent for suppressing the growth of a bacterium belonging to the genus Bylophila. More specifically, the present invention relates to a bacterium belonging to the genus Bilophila (Bilophila) and a method for inhibiting the growth of bacterium belonging to the genus Bylophila.

  Bilophila bacteria are a type of hydrogen sulfide-producing bacterium that produces hydrogen sulfide by anaerobic respiration in the intestine, suggesting that it is involved in inducing inflammation in the intestinal tract by destroying the intestinal mucosa. Is a kind of anaerobic pathogen.

  For example, in Non-Patent Document 1, an increase in the genus Bylophila was observed in the symptomatic score group in the ulcerative colitis model mouse (in particular, see FIG. 1). In established model mice, it has been reported that diets containing saturated fatty acids worsen the ulcer score. In Non-Patent Document 2, the proportion of the genus Bylophila in the appendix of a healthy person is 0.07 ± 0.06%, whereas the proportion of the genus Bylophila in the appendix of an appendicitis patient is 0.88 ±. It is reported that the rate increases by 0.2 times to about 10 times.

It has been known that there are various differences in the types and number of intestinal bacteria depending on the animal species and individual differences, the digestive tract site, age, diet content and physical condition, etc. In recent years, it has been found that the intestinal proportion changes depending on the diet style.
For example, Non-Patent Document 3 reports that the intestinal proportion of the genus Bylophila has been increased by a diet restricted to only meat and eggs.

  Recently, the style of Japanese meals has also become Western, and it has become a Western style with a focus on meat. For this reason, there is a concern that the intestinal proportion of the genus Vylophila, which is a kind of pathogenic bacteria, increases, and as a result, there is a possibility of inducing various digestive diseases.

By the way, as a method for suppressing the growth of bacteria of the genus Vylophila, a method of administering an antibiotic has been conventionally used.
For example, Patent Document 1 discloses that a chemifloxacin compound, which is a quinolone antibiotic, inhibits the growth of a bacterium belonging to the genus Vylophila.

Special table 2003-531101 gazette

Dietary-fat-induced taurocholic acid promotes pathobiontexpansion and colitis in Il10-/-mice. Nature 487, 104-108 (2012) Culture-Independent Evaluation of the Appendix and Rectum Microbiomes in Children with and without Appendicitis.PLoS One 4, e95414 (2014) Diet rapidly and reproducibly alters the human gut microbiome.Nature 505, 559-563 (2014)

  As described above, suppressing the growth of a bacterium belonging to the genus Vylophila, which is a kind of pathogenic bacteria, is very important in preventing various digestive diseases. Even in the prior art, the administration of antibiotics has shown a certain effect on the suppression of the growth of Vylophila bacteria. However, the administration of chemical substances has the risk of side effects, and may kill useful bacteria in the intestinal bacteria (so-called good bacteria), disturbing the balance of the intestinal microflora, etc. There are various problems.

  Therefore, the main object of the present technology is to provide a technology that suppresses the growth of the bacterium belonging to the genus Bylophila that is highly safe and does not adversely affect the balance of the intestinal flora.

  As a result of earnest research to solve the above problems, the present inventors have found that Bifidobacterium bacteria known as good bacteria contribute to the suppression of the growth of Vylophila bacteria. The technology has been completed.

That is, the present technology provides prevention of diseases caused by Bilophila wordsworthia (Bifophila worsworthia), which contains Bifidobacterium longumum ATCC BAA-999 as an active ingredient and proliferates with a meal centered on meat. / or Ru are used in the treatment, that provides Bairofira-Wazuwashia growth inhibitor.
A disease caused by Bilophila wordsworth that grows with meals centered on meat according to the present technology can include appendicitis.

In the present technology, Bifidobacterium longum ATCC BAA-999 is used as an active ingredient, and / or prevention of diseases caused by Bilophila wordsworthia that grows with meals centered on meat and / or Ru is used in the treatment, provides a Bairofira-Wazuwashia cytostatic food or drink composition.
Furthermore, in the present technology, Bifidobacterium longum ATCC BAA-999 is used as an active ingredient, and prevention of diseases caused by Bilophila wordsworthia that grows with meals centered on meat and / or Ru are used in the treatment, it provides a Bairofira-Wazuwashia cytostatic for beverage feed composition.

  According to this technology, Bifidobacterium genus bacteria that have been used for many years in foods and medicines and also exist in the intestines of animals are used as active ingredients, so that they are highly safe for living bodies and enteric bacteria. It is possible to provide a technique for suppressing the growth of a bacterium belonging to the genus Bylophila that does not adversely affect the balance of the flora. In addition, the effect described here is not necessarily limited, and may be any effect described in the present technology.

It is a figure which shows the test schedule in an Example. In an Example, it is a drawing substitute graph which shows a time-dependent change of the number of the bacteria of the genus Birophila in an intestinal microflora.

  Hereinafter, preferred embodiments for carrying out the present technology will be described. In addition, embodiment described below shows an example of typical embodiment of this technique, and, thereby, the scope of this technique is not interpreted narrowly.

<1. Virofila spp. Growth inhibitor and method of suppressing the growth of Vylophila spp>
The bacterium belonging to the genus Bylophila and the method for inhibiting the growth according to the present technology are characterized in that the bacterium belonging to the genus Bifidobacterium is an active ingredient.

(1) Bifidobacterium spp. Bifidobacterium spp. (Hereinafter simply referred to as “Bifidobacteria”) used as an active ingredient of the Byrophylla spp. Growth inhibitor according to the present technology is Actinobacteria spp. Bifidobacterium is a bacterium belonging to the genus Bifidobacteriaceae (Bifidobacterium) and is a kind of intestinal bacteria that inhabit the intestines of many animals.

  The type of Bifidobacterium that can be used in the growth inhibitor of the genus Bylophila according to the present technology should be selected from known Bifidobacterium as long as the effect of the present technology is not impaired. Can do. For example, Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium biscentibacterium (Bifidobacterium bifidum) Um Infantitis (Bifidobacterium infantis) and the like.

Among these, it is preferable to use Bifidobacterium longum (Bifidobacterium longum) in this technique. Bifidobacterium longum is one of the Bifidobacteria suitably used in foods such as fermented milk, and is also attracting attention from the viewpoint of improving immunity.
In the present technology, a strain known as Bifidobacterium longum can be appropriately used. More specifically, Bifidobacterium longum ATCC BAA-999 can be preferably used. The strain can be purchased from, for example, the American Type Culture Collection (address 12301 Parklawn Drive, Rockville, Maryland 20852, United States of America).

The content of the Bifidobacterium genus in the bacterium belonging to the genus Bylophila according to the present technology is not particularly limited, depending on the frequency of use, the amount used, the content and amount of the intake meal, the age of the intake, etc. Can be set freely. In the present technology, in particular, the content of the Bifidobacterium genus in the growth inhibitor of the genus Bylophila is preferably 1 × 10 ⁷ to 1 × 10 ¹² CFU / mL, and 4 × 10 ⁹ to 1 × 10 6. More preferably, it is ¹¹ CFU / mL. The unit CFU is an abbreviation for colony forming units and is a colony forming unit.

(2) Bylophila bacterium As a bacterium belonging to the genus Bylophila that can be inhibited by the growth inhibitor of the bacterium belonging to the genus Bilophila according to the present technology, there is a Bilophila wordsworthia species. Examples of the Bylophila bacterium belonging to the genus Bylophila Wordsworthia include Vylophila Wordsworthia ATCC 49260, Bylophila Wordsworthia ATCC 51580, Bilophila Wordsworthia ATCC 51581, and the like.

  The above-described inhibitor of the growth of the bacterium belonging to the genus Bilophila according to the present technology uses a Bifidobacterium bacterium that has been used for many years in foods and medicines and is also present in the intestines of animals as an active ingredient. High safety. Therefore, there is no risk of side effects and dependence, and it can be taken continuously for a long time.

  Bifidobacterium is a kind of bacteria that exist as so-called good bacteria in the intestines of animals. Therefore, the bacterium belonging to the genus Bylophila according to the present technology does not induce disorder in the balance of the intestinal microflora, but rather can be expected to have a balance adjusting action for the intestinal microflora.

<2. Drugs for preventing or treating diseases caused by the growth of the bacteria of the genus Bylophila>
As described above, the bacterium belonging to the genus Bylophila according to the present technology is highly safe and has no adverse effect on the balance of the intestinal microflora. It can be used as a pharmaceutical for humans or animals.

  In addition, the bacterium belonging to the genus Bylophila according to the present technology, which has been used in food for many years and has Bifidobacterium bacterium present in the intestines of animals as an active ingredient, has suffered from various diseases. There is a high possibility that it can be administered safely to patients. Moreover, there is little need to worry about side effects even if administered continuously for a long period of time. Furthermore, it is highly safe when used in combination with other drugs.

  In the case where the virulophila bacterial growth inhibitor according to the present technology is used as a pharmaceutical product, the pharmaceutical product may be administered either orally or parenterally, but oral administration is preferred. Examples of parenteral administration include rectal administration. Examples of the dosage form for oral administration include tablets, capsules, troches, syrups, granules, powders and the like.

  In addition to the Bifidobacterium genus, components such as excipients, pH adjusters, colorants, and corrigents that are commonly used for formulation can be used for formulation. In addition, as long as it contains an agent for suppressing the growth of a bacterium belonging to the genus Bilophylla according to the present technology, a component having an effect of preventing or treating a disease caused by the growth of a known bacterium belonging to the genus Virophila, It can also be used in combination with a bacterium belonging to the genus Bylophila according to the present technology.

  Furthermore, depending on the administration method, it can be appropriately formulated into a desired dosage form. For example, in the case of oral administration, it can be formulated into solid preparations such as powders, granules, tablets and capsules; liquid preparations such as solutions, syrups, suspensions and emulsions. For parenteral administration, it can be formulated into suppositories, ointments and the like.

  In addition, formulation can be appropriately performed by a known method according to the dosage form. In the formulation, only the Bifidobacterium bacterium that is an active ingredient may be formulated, or a formulation carrier may be blended as appropriate.

In addition, when a pharmaceutical carrier is blended, the content of the Bifidobacterium genus which is an active ingredient of the bacterium belonging to the genus Bylophila genus is not particularly limited, and the intake or dose per day according to the dosage form is not limited. Based on this, it can be selected as appropriate. The daily intake or dose of Bifidobacterium is preferably in the range of 1 × 10 ⁸ CFU / day to 1 × 10 ¹² CFU / day.

  In addition, as the preparation carrier, various organic or inorganic carriers can be used depending on the dosage form. Examples of the carrier in the case of a solid preparation include excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents.

  Examples of the excipient include sugar derivatives such as lactose, sucrose, glucose, mannitol and sorbit; starch derivatives such as corn starch, potato starch, α-starch, dextrin and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Cellulose derivatives such as hydroxypropylmethylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium magnesium magnesium silicate; phosphate derivatives such as calcium phosphate; And carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate and the like.

  Examples of the binder include gelatin, polyvinyl pyrrolidone, macrogol and the like in addition to the above excipients.

  Examples of the disintegrant include, in addition to the above excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone.

  As the lubricant, for example, talc; stearic acid; stearic acid metal salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and geirow; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid Carboxylic acid sodium salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as anhydrous silicic acid and silicic acid hydrate; starch derivatives and the like It is done.

  Examples of the stabilizer include paraoxybenzoates such as methyl paraben and propyl paraben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid and the like.

Examples of the flavoring agent include sweeteners, acidulants, and fragrances.
In addition, as a carrier used in the case of a liquid for oral administration, a solvent such as water, a flavoring agent and the like can be mentioned.

<3. Food and drink, feed>
As described above, the bacterium belonging to the genus Bylophila according to the present technology is highly safe and has no adverse effect on the balance of the intestinal microflora. It can be used by blending in human or animal food or drink, feed and the like.

(1) Food / beverage products When using the phyllophila bacterium growth inhibitor which concerns on this technique for food / beverage products, it can also add to well-known food / beverage products and can prepare the food / beverage products which have a phyllophila bacterium proliferation inhibitory effect, It is also possible to produce a new food or drink having an effect of suppressing the growth of the bacterium belonging to the genus Bylophila by mixing in the raw material of the product.

  The foods and drinks may be in the form of liquids, pastes, solids, powders, etc., in addition to tablet confections, liquid foods, feeds (including for pets), etc., for example, flour products, instant foods, processed agricultural products, marine products Processed products, processed livestock products, milk / dairy products, fats and oils, basic seasonings, compound seasonings / foods, frozen foods, confectionery, beverages, and other commercial products.

Examples of flour products include bread, macaroni, spaghetti, noodles, cake mix, fried flour, bread crumbs and the like.
Examples of instant foods include instant noodles, cup noodles, retort / cooked food, cooked canned food, microwave food, instant soup / stew, instant miso soup / soup, canned soup, freeze-dried food, other instant foods, etc. It is done.
Examples of processed agricultural products include canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, cereals (cereal processed products), and the like.
Examples of processed fishery products include canned fishery products, fish hams and sausages, marine products, marine delicacies, and tsukudani.
Examples of livestock processed products include canned livestock, pastes, livestock meat ham, sausage and the like.
Examples of milk / dairy products include processed milk, milk beverages, yogurts, lactic acid bacteria beverages, cheese, ice creams, prepared powdered milks, creams, and other dairy products.
Examples of the fats and oils include butter, margarine, vegetable oil and the like.
Basic seasonings include, for example, soy sauce, miso, sauces, tomato processed seasonings, mirins, vinegars, etc., and the above mixed seasonings and foods include cooking mix, curry ingredients, sauces, Examples include dressings, noodle soups, spices, and other complex seasonings.
Examples of the frozen food include raw material frozen food, semi-cooked frozen food, cooked frozen food, and the like.
Examples of the confectionery include caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pie, snacks, crackers, Japanese confectionery, rice confectionery, bean confectionery, dessert confectionery, and other confectionery.
Examples of beverages include carbonated beverages, natural fruit juices, fruit juice drinks, soft drinks with fruit juice, fruit drinks, fruit drinks with fruits, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks Sports drinks, nutritional drinks, alcoholic drinks, other taste drinks, and the like.
Examples of commercially available foods other than the above include baby foods, sprinkles, and tea paste.

In addition, the food and drink defined by the present technology can be provided and sold as a food or drink displaying health uses such as prevention or treatment of diseases caused by the growth of bacteria of the genus Bylophila.
The “display” act includes all acts for informing the consumer of the use, and if the expression can remind the user of the use, the purpose of the display, the content of the display, the display Regardless of the target object / medium, etc., all fall under the “display” act of this technology.

  Moreover, it is preferable that the “display” is performed by an expression that allows the consumer to directly recognize the above-described use. Specifically, it is the act of transferring, displaying, importing, displaying, or importing products that are related to food or drinks or products that describe the use, on advertisements, price lists, or transaction documents. For example, an act of describing and displaying the above uses or distributing them, or describing the above uses in information including the contents and providing them by an electromagnetic (Internet or the like) method can be given.

  On the other hand, the display content is preferably a display approved by the government or the like (for example, a display that is approved based on various systems determined by the government and performed in a mode based on such approval). Moreover, it is preferable to attach such display contents to advertising materials at sales sites such as packaging, containers, catalogs, pamphlets, POPs, and other documents.

  “Indication” also includes indication as health food, functional food, enteral nutrition food, special purpose food, health functional food, food for specified health, functional nutrition food, quasi drug, etc. Among these, in particular, there are indications approved by the Consumer Affairs Agency, for example, indications approved under the food system for specific health use, a similar system, and the like. Examples of the latter can include indications for food for specified health use, indications for conditional health foods, indications that affect the structure and function of the body, indications for reducing disease risk, etc. Specifically, indications as food for specified health (especially indications for health use) and similar indications stipulated in the Enforcement Regulations of the Health Promotion Act (Ministry of Health, Labor and Welfare Ordinance No. 86 of April 30, 2003) Is a typical example.

The content of the Bifidobacterium bacterium, which is an active ingredient of the growth inhibitor of the genus Bylophila added when producing food and drink, is not particularly limited and should be appropriately selected based on the daily intake. Can do. The daily intake of Bifidobacterium is preferably in the range of 1 × 10 ⁸ CFU / day to 1 × 10 ¹² CFU / day.

(2) Feed In the case where the virulophila bacterial growth inhibitor according to the present technology is used for feed, it can be added to a known feed to prepare a feed having an inhibitory effect on the growth of bacterial genus bacteria, It can also be mixed to produce a new feed having an inhibitory effect on the growth of the genus Bylophila.

  Examples of the raw material of the feed include cereals such as corn, wheat, barley, and rye; bran such as bran, wheat straw, rice bran, and defatted rice bran; and manufactured porridges such as corn gluten meal and corn jam meal; Animal feeds such as whey, fish meal and bone meal; yeasts such as beer yeast; mineral feeds such as calcium phosphate and calcium carbonate; fats and oils; amino acids; In addition, examples of the form of the feed include pet animal feed (pet food, etc.), livestock feed, fish feed, and the like.

In addition, the content of Bifidobacterium which is an active ingredient of the bacterium belonging to the genus Bylophila to be added when producing the feed is not particularly limited, and may be appropriately selected based on the daily intake. it can. The daily intake of Bifidobacterium is preferably in the range of 1 × 10 ⁸ CFU / day to 1 × 10 ¹² CFU / day.

  Hereinafter, the present invention will be described in more detail based on examples. In addition, the Example demonstrated below shows an example of the typical Example of this invention, and, thereby, the range of this invention is not interpreted narrowly.

(1) Test schedule 10 subjects who had 21 subjects (22-50 years old, average age: 39.5 years old) who took a meal of meat and eggs only (hereinafter referred to as “carnivorous”) for 5 days, and 5 Fermented milk containing Bifidobacterium longum ATCC BAA-999 at the same time as meat eating (manufactured by Morinaga Milk Industry Co., Ltd., Bihidas: registered trademark, Bifidobacterium longum ATCC BAA-999: about 2 × 10 ⁷ CFU / g)) was divided into two groups of 11 people taking 100 g twice a day in the morning and evening (see FIG. 1). The menu during the meat eating period and the post-observation period was as shown in Table 1 below. In addition, during the meat eating period, ingestion of either pattern 1 or pattern 2 shown in the following Table 1 was performed, and during the post-observation period, ingestion of a frozen lunch consisting of the menu shown in Table 1 below was performed.

[Carnivorous period]
Breakfast: Egg dishes (one egg) + processed meat (100g)
Lunch / Dinner: Main meal (150-200 g) + supplementary meal (80-100 g), depending on physique, gender, etc., main meal (150-200 g) was also possible. When supplementary meals were the main ingredient, 150 to 200 g or more was ingested.

(2) Analysis of gut microbiota During the carnivorous period of 5 days (1st to 5th day), before ingestion start (day 0), 3rd day, 3rd day, 5th day, 14 days ( During the post-observation period (from day 6 to day 20), fecal samples were collected from each subject on 3rd day, 8th day, 12th day, and 19th day, and the intestinal flora was analyzed. . The method for analyzing the intestinal microflora was as follows.

<Extraction of DNA>
DNA was extracted from a stool sample collected from the subject using QIA amp DNA tool Kit (Qiagen).
About 20 mg of a stool sample was weighed and suspended in 600 μL of ASL buffer, and then heated at 70 ° C. for 5 minutes. Here, 700 mg of 0.1 mm diameter glass beads and 500 μL of TE saturated phenol (manufactured by Wako Pure Chemical Industries, Ltd.) are mixed, and crushing is performed at 2700 rpm for 180 seconds using a multi-bead shocker (manufactured by Yasui Kikai Co., Ltd.). did. To this, 100 μL of 3 mol / L sodium acetate was added and left on ice for 5 minutes. After centrifugation at 14,000 rpm for 8 minutes, 600 μL of the supernatant was mixed with 600 μL of ASL buffer and 1 tablet of InhibitEX tablet, and the mixture was centrifuged at 14000 rpm for 2 minutes to obtain 471 μL of the supernatant. To this, 471 μL of AL buffer and 471 μL of ethanol solution (manufactured by Wako Pure Chemical Industries, Ltd.) were mixed, applied to QIA amp Spin Column, and centrifuged at 14000 rpm for 1 minute. The filtrate was discarded, 500 μL of AW1 buffer was applied to QIA amp Spin Column, and centrifuged at 14000 rpm for 1 minute. The filtrate was discarded, 500 μL of AW2 buffer was applied to QIA amp Spin Column, and centrifuged at 14000 rpm for 3 minutes. Furthermore, it centrifuged at 14000 rpm for 1 minute only with QIA amp Spin Column. This was transferred to a 1.5 mL tube, and 171 μL of AE buffer was added, left at room temperature for 1 minute, and then centrifuged at 14000 rpm for 1 minute to obtain a DNA solution for each sample.

<Amplification of DNA>
Next, a nucleic acid amplification reaction was performed on the genome of the fungal group contained in the DNA solution of each sample. Tables 2 to 4 below show the primer sequences used in this experimental example. The primer set shown in Table 2 is a primer for amplifying the 3rd to 4th variable regions of the 16S rRNA gene. These primers are referred to as the first primer set for convenience.

  In Table 2, “R” means guanine or adenine. “W” means adenine or thymine or uracil. “H” means adenine or cytosine or thymine or uracil. “V” means adenine or guanine or cytosine.

  In addition, the primer sets shown in Table 3 and Table 4 below are sequences (adapter regions and index regions) necessary for analyzing a base sequence using a next-generation sequencer (Miseq (manufactured by Illumina)). It is a primer set for incorporation. The primers shown in Table 3 are forward primers, and the primers shown in Table 4 are reverse primers. These primers are referred to as the second primer set for convenience. The primers shown in Tables 2 to 4 were obtained by Life Technologies' oligo primer preparation service.

  A reaction solution having a total volume of 25 μl containing the DNA solution of each sample and the first primer set was prepared using TaKaRa ExTaq HS kit (Takara Bio Inc.). The nucleic acid amplification reaction is performed using a thermal cycler (Veriti 200 (manufactured by Life Technologies)), 94 ° C. for 3 minutes, 94 ° C. for 30 seconds, 50 ° C. for 30 seconds, and 72 ° C. for 30 seconds for one cycle. This was repeated 20 times, and then the PCR reaction was carried out at 72 ° C. for 10 minutes. The obtained PCR product was electrophoresed on a 1% agarose gel, and the amplified nucleic acid chain was confirmed by the band pattern.

  Subsequently, 1 μL of the obtained PCR product was used as a template, and a PCR reaction was performed using the second primer set under the same conditions as described above. However, the number of cycles was eight. The obtained PCR product was electrophoresed on a 1% agarose gel, and after confirming the band pattern, the PCR product was purified using GeneRead Size Selection Kit (manufactured by Qiagen). For the purified PCR product, the DNA concentration was measured by Quant-iT PicoGreen dsDNA Assay kit (manufactured by Life Technologies).

<DNA sequence>
A mixture of PCR products derived from each sample at the same concentration was subjected to Miseq v2 Reagent kit (manufactured by Illumina), and sequence analysis was performed with Miseq.

<Identification of bacteria group and calculation of the number of bacteria>
The paired end sequence obtained by the above sequence analysis is transferred to the QIIME software (version 1.1.2-301) (http://code.google.com/p/ea-utils/wiki/FastqJoin) 1.6.0) (http://qiime.org/), each sequence having 97% homology was defined as OUT (Operational Taxonomy Unit). The structure of the intestinal flora was analyzed by BLAST search of the representative sequence of each OUT against the following database.
Database: Greengenes database 12_10
(http://greengenes.secondgenome.com/downloads/database/12_10)

(3) Results FIG. 2 shows time-dependent changes in the number of the genus Bylophila in the intestinal flora obtained by the analysis of the intestinal flora. As shown in the results of FIG. 2, in the carnivorous-only group, a significant increase in the genus Bylophila was evident during the carnivorous period. On the other hand, in the group that ingested fermented milk (Bihidas) containing Bifidobacterium bacteria at the same time as the meat diet, no increase in Birophylla bacteria was observed even during the carnivorous period.

  From the above results, it can be seen that Bifidobacterium has an action of suppressing the increase of Byrophylla bacteria, and that Bifidobacterium is also in the diet of carnivorous food that promotes the increase of Biliophila bacteria. It was proved that ingestion of the genus bacteria can suppress the increase in the genus Bylophila.

Claims (4)

Bifidobacterium longum Uses ATCC BAA-999 as an active ingredient, and is used for prevention and / or treatment of diseases caused by Bilophila wordsworthia that grows with meat-centered meals It is Ru, Bairofira-Wazuwashia growth inhibitor. The virophila wordsworthia growth inhibitor according to claim 1, wherein the disease caused by bilophila wordsworthia that grows with meals centered on meat is appendicitis. Bifidobacterium longum Uses ATCC BAA-999 as an active ingredient, and is used for prevention and / or treatment of diseases caused by Bilophila wordsworthia that grows with meat-centered meals is Ru, Bairofira-Wazuwashia growth inhibition for food and drink composition. Bifidobacterium longum Uses ATCC BAA-999 as an active ingredient, and is used for prevention and / or treatment of diseases caused by Bilophila wordsworthia that grows with meat-centered meals It is Ru, Bairofira-Wazuwashia cytostatic feed composition.

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