HK1181314B - Prophylactic/ameliorating agent for diseases, endurance improving agent and anti-fatigue agent, and pharmaceutical agent, food and beverage each comprising any one of those agents - Google Patents

Description

Prophylactic/ameliorating agent for disease, endurance improver, anti-fatigue agent, and pharmaceutical and food/drink using same

Technical Field

The present invention relates to an agent for preventing and ameliorating a disease, an agent for improving endurance, an anti-fatigue agent, and a pharmaceutical product and a food or drink each using the same.

Background

It is known that the homeostasis of a living body is important for the balance of networks such as the immune system, the nervous system, and the endocrine system. One of the causes of diseases such as inflammatory diseases, allergic diseases, autoimmune diseases, cancer, and infectious diseases is considered to be disorder of the network balance. In addition, the fatigue includes various types, including fatigue accompanied by serious diseases such as malignant tumor, infectious disease, chronic fatigue syndrome, and the like, and usual fatigue derived from mental and physical stress in daily life.

Conventionally, many agents for preventing and ameliorating inflammatory diseases, allergic diseases, autoimmune diseases, and the like, agents for improving endurance, and anti-fatigue agents have been difficult to use for a long period of time because they have side effects as well as effects. Therefore, in view of the consideration of both effectiveness and safety, a preventive and ameliorating agent using Pleurotus nebrodensis (Pleurotus nebrodensis) has been reported (see, for example, patent document 1).

On the other hand, there has been reported a health food using a metabolite produced by mixed culture of red photosynthetic bacteria and lactic acid bacteria in order to improve the health state of an unhealthy person (see patent document 2). The health food is excellent in efficacy and safety, but a prophylactic improver, a stamina improver and an anti-fatigue agent which further improve the performance are desired.

Documents of the prior art

Patent document

Patent document 1: japanese laid-open patent publication No. 2006-241115

Patent document 2: international publication No. 01/60977

Disclosure of Invention

Problems to be solved by the invention

The purpose of the present invention is to provide a prophylactic/ameliorating agent, a durability-enhancing agent, and an anti-fatigue agent that are useful for the prevention and amelioration of inflammatory diseases, allergic diseases, and autoimmune diseases and that are highly safe.

Means for solving the problems

In order to achieve the above object, the prophylactic/ameliorating agent of the present invention is characterized by comprising at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, and at least one of red non-sulfur bacteria containing nitrogen-fixing red bacteria (Rhodobacter azotoformans) and a culture of the red non-sulfur bacteria.

The endurance improver and the anti-fatigue agent of the present invention are characterized by containing at least one of red non-sulfur bacteria containing nitrogen-fixing red bacteria (Rhodobacter azotoformans) and a culture of the red non-sulfur bacteria.

ADVANTAGEOUS EFFECTS OF INVENTION

In order to achieve the above object, the present inventors have repeatedly conducted a series of studies and, as a result, have found a novel red non-sulfur bacterium having the following activity: the present invention has been completed based on the finding that the present invention has an inhibitory activity on ulcer, an inhibitory activity on inflammatory cell infiltration, an inhibitory activity on swelling of mucosal epithelium, epithelial regeneration, a reduction in serum IgE and histamine levels, an inhibitory activity on Th 2-type cytokine secretion, an improvement in Th1/Th2 imbalance, an inhibitory activity on allergic reactions, an inhibitory activity on autoimmune diseases, an improvement in endurance, an anti-fatigue activity, and the like. The red non-sulfur bacterium of the present invention is useful for prevention and improvement, endurance improvement, and fatigue resistance of at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases. In addition, since the red non-sulfur bacterium of the present invention is highly safe, it can be administered for the purpose of prevention before the disease occurs, and can be administered for a long period of time.

Drawings

FIG. 1 is a photograph of pathological tissues of a colon tissue section in example 2. Fig. 1 (a) is a photograph of the pathological tissue of example 2 (magnification: 32 ×), and fig. 1 (B) is a photograph of the pathological tissue of example 2 (magnification: 80 ×).

FIG. 2 is a photograph of pathological tissues of a colon tissue section in comparative example 1. Fig. 2 (a) is a photograph of the pathological tissue of comparative example 1 (magnification: 32 ×), and fig. 2 (B) is a photograph of the pathological tissue of comparative example 1 (magnification: 80 ×).

FIG. 3 is a photograph of pathological tissues of a colon tissue section in comparative example 2. Fig. 3 (a) is a photograph of the pathological tissue of comparative example 2 (magnification: 32 ×), and fig. 3 (B) is a photograph of the pathological tissue of comparative example 2 (magnification: 80 ×).

FIG. 4 is a graph showing the average values of edema volumes in example 8, comparative example 8-1 and comparative example 8-2.

FIG. 5 is a graph showing the average values of edema volumes in example 9, comparative example 9-1 and comparative example 9-2.

FIG. 6 is a soft X-ray photograph of the hind limb of example 9, comparative example 9-1 and comparative example 9-2.

FIG. 7 is a graph showing the average values of the amounts of histamine in sera in example 10, comparative example 10-1 and comparative example 10-2.

FIG. 8 is a graph showing the measurement results of the drape method in example 11.

FIG. 9 is a graph showing the measurement results of the drape method in example 12.

Detailed Description

Among the prevention improver, endurance improver and anti-fatigue agent of the present invention, Rhodobacter azotoformans (Rhodobacter azotoformans) preferably have the following bacteriological characteristics (1) to (30). Examples of the bacteria having such bacteriological characteristics include Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain. The BP0899 strain was deposited under the accession number NITE P-644 (deposition date: 2008, 9/12) at the International patent organism depositary, national institute of technology and evaluation, basic disk organization for evaluation of products, Japan (2-5-8 of Total Sickle foot, Otsumadzu, Kyowa prefecture, Japan), and further deposited under the accession number NITE BP-644 (shift date: 2010, 10/27).

(1) Shape of the cells: rod-like or ovoid shape

(2) Polymorphism: is free of

(3) Size of the cells: 0.8 μm × 1.0 μm

(4) Presence or absence of motility: is provided with

(5) Presence or absence of spores: is free of

(6) Gloss on regular agar culture: is provided with

(7) Pigment production on normal agar culture: is provided with

(8) Presence or absence of surface development in ordinary broth culture: is free of

(9) Presence or absence of turbidity of the medium in the case of ordinary broth culture: is provided with

(10) Liquefaction of gelatin during gelatin puncture culture: negative of

(11) Solidification of litmus milk during culture: is free of

(12) Liquefaction of litmus milk during culture: is free of

(13) Gram staining property: negative of

(14) Reduction of nitrate: is free of

(15) And (3) denitrification reaction: is provided with

(16) And (3) MR test: negative of

(17) Indole production: is free of

(18) Generation of hydrogen sulfide: is free of

(19) Hydrolysis of starch: is free of

(20) Use of citric acid (Christensen (k)): is free of

(21) Utilization of inorganic nitrogen source (ammonium salt): is provided with

(22) Production of Catalase: positive for

(23) Production of oxidase: positive for

(24) Anaerobic fertility: is provided with

(25) O-F test (oxidation/fermentation): negative/negative

(26) β -galactosidase activity: negative of

(27) Arginine double hydrolase activity: negative of

(28) Lysine decarboxylase activity: negative of

(29) Tryptophan deaminase activity: negative of

(30) Gelatinase activity: negative of

In the prophylactic and ameliorating agent, the endurance improver and the anti-fatigue agent of the present invention, the base sequence of 16S rRNA of Rhodobacter azotoformans (Rhodobacter azotoformans) is preferably the base sequence shown in sequence No. 1.

The first pharmaceutical product of the present invention is a pharmaceutical product for preventing or treating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, and contains the prophylactic/ameliorating agent of the present invention.

The second pharmaceutical product of the present invention is characterized by containing at least one of the endurance improving agent and the anti-fatigue agent of the present invention.

The first food or drink of the present invention is a food or drink having a function of preventing or ameliorating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, and is characterized by containing the prophylactic/ameliorating agent of the present invention.

The second food or drink of the present invention is characterized by containing at least one of the endurance improving agent and the anti-fatigue agent of the present invention.

The red non-sulfur bacterium of the present invention is characterized by being Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain (accession No. NITE BP-644).

The method for preventing and ameliorating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, according to the present invention is characterized by comprising a step of administering a preventive ameliorating agent comprising at least one of red non-sulfur bacteria and a culture of the red non-sulfur bacteria, wherein the red non-sulfur bacteria comprise nitrogen-fixing red bacteria (Rhodobacter azotoformans).

The method for improving endurance of the present invention is characterized by comprising a step of administering an endurance improving agent containing at least one of red non-sulfur bacteria containing nitrogen-fixing red bacteria (rhodobacter azotoformans) and a culture of the red non-sulfur bacteria.

The method for relieving fatigue of the present invention is characterized by comprising a step of administering an anti-fatigue agent containing at least one of red non-sulfur bacteria containing nitrogen-fixing red bacteria (rhodobacter azotoformans) and a culture of the red non-sulfur bacteria.

The first red non-sulfur bacteria (rhodobacter azotoformans) or the culture thereof of the present invention are: red non-sulfur bacteria (Rhodobacter azotoformans) or a culture thereof for use in a method for preventing or ameliorating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases.

The second red non-sulfur bacterium (rhodobacter azotoformans) or the culture thereof of the present invention is: a red non-sulfur bacterium (rhodobacter azotoformans) or a culture thereof for use in at least one of the endurance improving method and the anti-fatigue method.

The first use of the invention is: use of red non-sulfur bacteria (Rhodobacter azotoformans) or a culture thereof for the prophylactic improvement of at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases.

The second application of the invention is as follows: use of red non-sulfur bacteria (Rhodobacter azotoformans) or a culture thereof for at least one of endurance improvement and fatigue resistance.

The present invention is described in detail below.

In the present invention, the improvement of the endurance includes, for example, suppressing a decrease in the endurance of the body against the load, improving the endurance of the body against the load, and the like. The endurance may be, for example, endurance at the time of performing each activity, and specifically, endurance at the time of performing activities such as labor and sports, endurance at the time of performing each activity of daily life, and the like.

In the present invention, antifatigue means, for example, prevention, alleviation, recovery or improvement of fatigue symptoms. Examples of the fatigue include fatigue due to activities such as labor and physical exercise, fatigue due to activities in daily life, and fatigue due to diseases. The aforementioned fatigue may include, for example, a feeling of burnout.

In the present invention, the improvement of the endurance and the fatigue resistance may be referred to as improvement of physical strength, for example. Specific examples of the improvement in physical strength include maintenance, reduction inhibition, and improvement of physical strength.

In the present invention, the aforementioned endurance improving and antifatigue effects can be attributed to, for example, an anti-inflammatory effect brought about by the aforementioned red non-sulfur bacteria. The anti-inflammatory effect can be achieved by, for example, inhibiting the expression of inflammatory cytokines using the red non-sulfur bacterium. The inflammatory cytokine is not particularly limited, and examples thereof include IL-1 and IFN.

Conventional anti-fatigue agents generally have a high factor such as nutritional supplementation. On the other hand, for example, the endurance builder and the anti-fatigue agent of the present invention may be a preparation in which a fatigue-causing substance is adjusted from an immunological approach of anti-inflammation by tightly grasping fatigue as a ring of inflammatory symptoms.

The prophylactic/ameliorating agent of the present invention is characterized by containing at least one of red non-sulfur bacteria containing nitrogen-fixing red bacteria (Rhodobacter azotoformans) and a culture of the red non-sulfur bacteria, as described above, and is a prophylactic/ameliorating agent for at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases. The endurance improver and the anti-fatigue agent of the present invention are characterized by comprising at least one of the red non-sulfur bacteria containing nitrogen-fixing red bacteria (Rhodobacter azotoformans) and the culture of the red non-sulfur bacteria, as described above.

< bacterium >

In the present invention, the aforementioned azotobacteria (Rhodobacter azotoformans) belonging to the aforementioned red non-sulfur bacteria preferably have the bacteriological characteristics of the following (1) to (30) as described above. The aforementioned azotobacteria (Rhodobacter azotoformans) having such bacteriological characteristics may be referred to as, for example, the aforementioned red non-sulfur bacteria of the present invention. Specific examples of the bacteria having such bacteriological characteristics include, for example, Rhodobacter azotoformans BP0899 (accession No. NITE BP-644) as described above.

(1) Shape of the cells: rod-like or ovoid shape

(2) Polymorphism: is free of

(3) Size of the cells: 0.8 μm × 1.0 μm

(4) Presence or absence of motility: is provided with

(5) Presence or absence of spores: is free of

(6) Gloss on regular agar culture: is provided with

(7) Pigment production on normal agar culture: is provided with

(8) Presence or absence of surface development in ordinary broth culture: is free of

(9) Presence or absence of turbidity of the medium in the case of ordinary broth culture: is provided with

(10) Liquefaction of gelatin during gelatin puncture culture: negative of

(11) Solidification of litmus milk during culture: is free of

(12) Liquefaction of litmus milk during culture: is free of

(13) Gram staining property: negative of

(14) Reduction of nitrate: is free of

(15) And (3) denitrification reaction: is provided with

(16) And (3) MR test: negative of

(17) Indole production: is free of

(18) Generation of hydrogen sulfide: is free of

(19) Hydrolysis of starch: is free of

(20) Utilization of citric acid (Christensen): is free of

(21) Utilization of inorganic nitrogen source (ammonium salt): is provided with

(22) Production of Catalase: positive for

(23) Production of oxidase: positive for

(24) Anaerobic fertility: is provided with

(25) O-F test (oxidation/fermentation): negative/negative

(26) β -galactosidase activity: negative of

(27) Arginine double hydrolase activity: negative of

(28) Lysine decarboxylase activity: negative of

(29) Tryptophan deaminase activity: negative of

(30) Gelatinase activity: negative of

The red non-sulfur bacteria may further exhibit the properties shown in the following table (31), for example, under aerobic culture conditions in a dark place. In the table, "-" indicates no production, and "+" indicates production.

(31) Acid and gas production based on sugars

The bacteriological characteristics can be evaluated, for example, from the results obtained by performing the present culture after the preculture. The preculture can be carried out, for example, by inoculating the red non-sulfur bacterium in a common agar medium and culturing the bacterium at 30 ℃ for 24 hours. The conditions for the present culture can be appropriately set according to the evaluation method of each bacteriological characteristic. Specifically, the culture conditions of (1) to (5) are, for example, aerobic culture at 30 ℃ in the dark using a common agar medium, the culture conditions of (6) to (7) are, for example, anaerobic culture at 30 ℃ in the light using a common broth medium, the culture conditions of (8) to (12) are, for example, aerobic culture at 30 ℃ in the dark using each medium, the oxidation tests of (13), (14), (16), (17), (19) to (23), (25), (26), (29), (30) and (31) are, for example, aerobic culture in the dark, (15), (18), (24), (25) fermentation tests, and (27) and (28) are, for example, anaerobic culture in the dark. The method for testing the bacteriological characteristics is not particularly limited, and conventionally known methods can be used. Specifically, for example, Barrow G.I. and Feltham R.K.A. entitled "Cowan and Steel's Manual for the Identification of medical bacteria", 3rd edi (UK)A method described in "New bacteria culture Medium lecture & Settlement" (Tokyo) second edition nearly big publication 1988, Changchun's editions "Classification and identification of microorganisms" (Sekko) "academic Press published by Cambridge University Press 1993 and Sakazaki et al, 1985, soil microorganism research institute compiled" soil microbiology experiment "(Tokyo) Yanxiantang 1992. The test method (15) can be performed by, for example, the method described in "classification and identification of microorganisms (below)" as mentioned above, the method described in "newly compiled soil microbiology experiment" using Giltay medium, or the method using PYN medium for drainage. The aforementioned method of foal-shape or the like determines that the denitrification reaction is positive when growth and gas generation are confirmed under anaerobic culture conditions using 1% sodium nitrate gravy. The method using the Giltay medium judges that denitrification reaction is positive when gas is generated and the color is dark blue under anaerobic culture conditions using the Giltay medium (pH 7.0-7.2) inserted with a Dallam tube. The Giltay medium is a Giltay medium prepared by mixing solution A (KNO)₃1g, 1g of asparagine, 5mL of 1% bromothymol blue-alcohol solution and 500mL of distilled water) and solution B (8.5 g of sodium citrate, MgSO 2₄·7H₂O1g、FeCl₃-6H₂O0.05g、KH₂PO₄1g、CaCl₂-6H₂0.2g of O and 500mL of distilled water). In addition, the aforementioned test method may use, for example, a commercially available bacteria identification kit. The kit is not particularly limited, and for example, a bacteria identification kit API20E (manufactured by biomerieux corporation) or the like can be used.

The aforementioned Rhodobacter azotoformans (Rhodobacter azotoformans) may further have the following bacteriological properties (32) to (40), for example.

(32) Color of colony: red colour

(33) Gelatin puncture culture: birth defect

(34) VP test: negative of

(35) Citric acid utilization (Koser): is provided with

(36) Utilization of inorganic nitrogen source (nitrate): is provided with

(37) Urease activity: negative of

(38) pH range for breeding: 5 to 9

(39) Acid production based on D-mannitol: has the generation of

(40) Gas production based on D-mannitol: no generation of

The method for testing the bacteriological characteristics of the above-mentioned items (32) to (40) is not particularly limited, and conventionally known methods can be employed. Specifically, for example, the methods described in the above-mentioned documents and the like are mentioned. In addition, the aforementioned test method may use, for example, a commercially available bacteria identification kit. The kit is not particularly limited, and for example, the above-described bacteria identification kit and the like can be used.

The aforementioned red non-sulfur bacteria may further contain, for example, other red non-sulfur bacteria other than nitrogen-fixing red bacteria (Rhodobacter azotoformans).

The other red non-sulfur bacteria are not particularly limited, and examples thereof include bacteria of the genera Rhodospirillum, Rhodosporidium, Rhodococcus, Rhodomicrobium, Blastochlorsis, Rhodoplanus, Rhodococcus, Rhodotorula, Rhodopseudomonas, etc.

The other red non-sulfur bacteria may be bacteria of the genus Rhodobacter (Rhodobacter) other than the aforementioned Rhodobacter azotoformans, for example.

The collection source of the red non-sulfur bacteria is not particularly limited, and examples thereof include soil, seawater, river water, lake water, and marsh water. Examples of the soil include, but are not particularly limited to, soil on land, sea bottom, river bottom, lake bottom, and marsh bottom, sand, and soil.

The method for separating the red non-sulfur bacteria is not particularly limited, and conventionally known collection methods and culture methods can be used, for example. As the separation method, for example, when the collection source is lake water, the collected lake water is filtered with a filter or the like, the filtrate is cultured in an agar medium or the like, and the red non-sulfur bacteria are separated from the obtained colonies. For example, when the collection source is soil, the collected soil may be suspended in a buffer solution or the like, the suspension may be centrifuged, the obtained supernatant may be cultured in an agar medium or the like, and the red non-sulfur bacteria may be separated from the obtained colonies. The red non-sulfur bacterium isolated as described above may be further cultured in, for example, a liquid medium.

The preventive/ameliorating agent, endurance improver and anti-fatigue agent of the present invention may further contain other bacteria in addition to the red non-sulfur bacteria. The other bacteria are not particularly limited, and examples thereof include lactic acid bacteria, yeast and the like, and lactic acid bacteria are preferred.

The lactic acid bacteria include, but are not particularly limited to, Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), Lactobacillus lactis (Lactobacillus lactis), Lactobacillus bulgaricus (Lactobacillus bulgaricus), Lactobacillus helveticus (Lactobacillus helveticus), Lactobacillus delbrueckii (Lactobacillus delbrueckii), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus brevis (Lactobacillus brevis), Bifidobacterium longum (Bifidobacterium longum), Bifidobacterium breve (Bifidobacterium breve), Enterococcus faecalis (Enterococcus faecalis), Streptococcus thermophilus (Streptococcus thermophilus), Lactobacillus acidophilus (Streptococcus thermophilus), and the like, preferably Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus acidophilus (Lactobacillus), and the like.

The yeast is not particularly limited, and examples thereof include Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Saccharomyces ellipsoidea, and Saccharomyces rouxii.

The medium for culturing the above-mentioned red non-sulfur bacteria is not particularly limited, and examples thereof include a medium supplemented with a lower fatty acid, a medium supplemented with malic acid, an L-dry standard recovery medium 802 "DAIGO" (manufactured by Japan pharmaceuticals), a MYS medium (Pink Stone and Beichuan, Bulletin of the Japanese society of Scientific Fisheries, 1984, Vol.50, No. 11, p.1929-1937), a modified MYS medium, a growth medium, and the like, and preferably include a medium supplemented with a lower fatty acid, a medium supplemented with malic acid, and an L-dry standard recovery medium 802 "DAIGO" (manufactured by Japan pharmaceuticals).

Examples of the medium to which the lower fatty acid is added and the medium to which malic acid is added include a medium to which biotin, vitamin B1, nicotinic acid, a lower fatty acid, or a sodium salt of malic acid is added to a basal medium shown in table 1 below. The lower fatty acid is not particularly limited, and for example, acetic acid, propionic acid, lactic acid and the like are preferable.

TABLE 1

Examples of the modified MYS medium and the growth medium include media having the compositions shown in tables 2 and 3 below.

TABLE 2

TABLE 3

In the above culture, the temperature range is not particularly limited, and is, for example, 23 to 39 ℃ and preferably 30 ℃.

In the above culture, the pH range is not particularly limited, and is, for example, a pH range of 5.5 to 8.5, preferably a pH range of 6.0 to 8.5, and more preferably a pH of 7.0.

The culture may be carried out, for example, under aerobic conditions or under anaerobic conditions, but is not particularly limited, and preferably under anaerobic conditions. The light conditions during the culture are not particularly limited, and may be, for example, dark conditions or illumination conditions, and are preferably an illumination of 2000 lux to 10000 lux. The culture can be carried out, for example, in a closed illumination culture vessel. Further, the culture can be performed while stirring the culture solution by using a stirring device provided in the sealed illumination type culture tank.

The culture time is not particularly limited, and may be, for example, a time until the growth of the red non-sulfur bacteria reaches a stationary phase. When the growth of the red non-sulfur bacteria is performed under the culture conditions in which the growth reaches a stationary phase within about 72 hours, the culture time may be 72 hours, for example.

In the culture of the red non-sulfur bacteria, for example, only the red non-sulfur bacteria may be cultured, or other bacteria may be simultaneously cultured in a mixed manner. The other bacteria are not particularly limited, and examples thereof include the aforementioned lactic acid bacteria and yeast.

As described above, the 16S rRNA of the aforementioned Rhodobacter azotoformans (Rhodobacter azotoformans) preferably has the base sequence shown in SEQ ID NO. 1. Specific examples of the red non-sulfur bacterium having the base sequence of 16S rRNA include Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain (accession No. NITE BP-644).

The nucleotide sequence of the 16S rRNA can be determined, for example, by extracting DNA from the red non-sulfur bacteria isolated and cultured by the method described above, or the like, and using primers or the like. The method for extracting the DNA and measuring the nucleotide sequence may be, for example, a usual method, and is not particularly limited. The primer is not particularly limited, and examples thereof include the following primers.

(primers)

9F (SEQ ID NO. 2)

5’-GAGTTTGATCCTGGCTCAG-3’

339F (SEQ ID NO. 3)

5’-CTCCTACGGGAGGCAGCAG-3’

785F (SEQ ID NO. 4)

5’-GGATTAGATACCCTGGTAGTC-3’

1099F (SEQ ID NO. 5)

5’-GCAACGAGCGCAACCC-3’

536R (SEQ ID NO. 6)

5’-GTATTACCGCGGCTGCTG-3’

802R (SEQ ID NO. 7)

5’-TACCAGGGTATCTAATCC-3’

1242R (SEQ ID NO. 8)

5’-CCATTGTAGCACGTGT-3’

1541R (SEQ ID NO. 9)

5’-AAGGAGGTGATCCAGCC-3’

Rhodobacteria azotoformans (Rhodobacteria azotoformans) having the bacteriological properties of the above (1) to (30) or at least one of 16S rRNAs having a base sequence represented by SEQ ID NO. 1 can be referred to as the red non-sulfur bacteria of the present invention.

< culture >

Examples of the culture of the red non-sulfur bacterium include, but are not particularly limited to, bacterial cells of the red non-sulfur bacterium, a culture supernatant of the red non-sulfur bacterium, and an extract of bacterial cells of the red non-sulfur bacterium. The prophylactic or ameliorating agent, endurance improver and anti-fatigue agent of the present invention may further comprise a culture of a bacterium other than the red non-sulfur bacterium. The culture of the bacteria other than the red non-sulfur bacteria is not particularly limited, and examples thereof include cell bodies of the other bacteria, culture supernatants of the other bacteria, and cell extracts of the other bacteria. Specific examples of the culture of bacteria other than the red non-sulfur bacteria include dried cells and extracts of the aforementioned lactic acid bacteria and yeast.

The culture may be, for example, a treated product of the cell, a treated product of the culture supernatant, a treated product of the cell extract, or the like, and is not particularly limited. The treated product is not particularly limited, and examples thereof include a concentrate, a dried product, a freeze-dried product, a solvent-treated product, a surfactant-treated product, an enzyme-treated product, a protein-fractionated product, an ultrasonic-treated product, and a ground product of the culture. The culture may be a mixture of the cells, the culture supernatant, the cell extract, a treated cell, a treated culture supernatant, a treated cell extract, and the like. The mixture may be mixed in any combination and ratio, and is not particularly limited. The combination is not particularly limited, and examples thereof include a mixture of the cells and the culture supernatant.

The preventive/ameliorating agent, endurance improver and anti-fatigue agent of the present invention may further contain other components such as additives. The additive is not particularly limited, and examples thereof include a stabilizer. The production method of the prophylactic/ameliorating agent, the endurance improver and the anti-fatigue agent is not particularly limited, and for example, a generally used pharmaceutical preparation technique can be used.

As described above, the prophylactic/ameliorating agent of the present invention is a prophylactic/ameliorating agent for preventing and ameliorating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, and contains at least one of the red non-sulfur bacterium and a culture of the red non-sulfur bacterium. In the present invention, the term "prevention of improvement" means that the composition has at least one of the effects of prevention and improvement. Therefore, the prophylactic and ameliorating agents of the present invention include "prophylactic and ameliorating agents", "prophylactic agents" and "ameliorating agents".

The inflammatory disease is not particularly limited, and examples thereof include inflammatory diseases of the internal organs, skin, joints, central nervous system, and the like. Specific examples of the inflammatory disease include inflammatory bowel diseases such as ulcerative colitis and crohn's disease, inflammatory skin diseases such as psoriasis and dermatitis, encephalitis, hepatitis, nephritis, pneumonia, bronchitis, vasculitis, meningitis, thyroiditis, diabetes, inflammatory bile diseases, and cancers associated with inflammation, and are not particularly limited. By preventing and ameliorating the anti-inflammatory effect in the inflammatory disease, the analgesic effect of pain associated with inflammation is also exhibited.

The allergic disease is not particularly limited, and examples thereof include type I allergic disease, type II allergic disease, type III allergic disease, type IV allergic disease, type V allergic disease, and the like, and type I allergic disease or type IV allergic disease is preferable. The type I allergic disease is not particularly limited, and specific examples thereof include urticaria, pollinosis, asthma, PIE syndrome (eosinophilic lung infiltration), atopic dermatitis, allergic rhinitis, allergic conjunctivitis, food allergy, drug allergy, and anaphylaxis. The type IV allergic disease is not particularly limited, and specific examples thereof include contact dermatitis and the like.

Examples of the autoimmune disease include, but are not particularly limited to, rheumatoid arthritis, lupus erythematosus, multiple sclerosis, systemic sclerosis, psoriatic arthritis, infection, and sjogren's syndrome.

The prophylactic/ameliorating agent of the present invention may be used for preventing and ameliorating at least one disease selected from the group consisting of the inflammatory diseases, allergic diseases and autoimmune diseases, and may be used for preventing and ameliorating a plurality of diseases. In addition, the prophylactic and ameliorating agent of the present invention can further prevent and ameliorate other diseases. The other diseases are not particularly limited, and examples thereof include circulatory diseases, cancers, and infectious diseases.

The circulatory diseases are not particularly limited, and examples thereof include hypertension, myocardial infarction, angina pectoris, arteriosclerosis, and the like.

The cancer is not particularly limited, and examples thereof include lung cancer, liver cancer, gastrointestinal cancer, kidney cancer, pancreatic cancer, thyroid cancer, prostate cancer, ovarian cancer, uterine cancer, and bone cancer.

The infection is not particularly limited, and examples thereof include infection with various viruses such as hepatitis virus, influenza virus, and human immunodeficiency virus, and infection with MRSA (methicillin-resistant staphylococcus aureus), hemolytic streptococcus, and mycoplasma.

< medicine >

The first pharmaceutical product of the present invention is a pharmaceutical product for preventing or treating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, and is not limited at all except for containing the prophylactic/ameliorating agent of the present invention. The first pharmaceutical product of the present invention may further contain at least one of the endurance improving agent and the anti-fatigue agent of the present invention. The second pharmaceutical product of the present invention is not limited at all except for containing at least one of the endurance improving agent and the anti-fatigue agent of the present invention. The second pharmaceutical product of the present invention may further contain the prophylactic or ameliorating agent of the present invention. Hereinafter, the first medicine and the second medicine of the present invention may be collectively referred to as "medicines". In the present invention, the medicine includes a medicine and a quasi-medicine. The pharmaceutical composition of the present invention may further contain other prophylactic or ameliorating agents, endurance improving agents, and anti-fatigue agents. The other preventive/ameliorating agent is not particularly limited, and examples thereof include the preventive/ameliorating agents for other diseases described above. The other endurance improvers and anti-fatigue agents are not particularly limited.

The dosage form of the above-mentioned drug is not particularly limited, and examples thereof include powder, fine granules, tablets, coated tablets, capsules, troches, and liquids. The composition of the pharmaceutical product is not particularly limited, and the pharmaceutical product may contain various additives such as an excipient, a binder, a lubricant, a disintegrant, an absorption enhancer, an emulsifier, a stabilizer, and a preservative, in addition to the prevention improver, the endurance improver, and the anti-fatigue agent. The above-mentioned drug can be produced by a conventionally used formulation technique or the like. The animal species to which the above-mentioned drug is administered is not particularly limited, and examples thereof include humans, non-human mammals such as monkeys, cows, pigs, dogs, and cats, birds such as chickens, and fish and shellfish. The administration method is not particularly limited, and examples thereof include oral administration and parenteral administration, and examples thereof include transdermal absorption, injection, and suppository administration. The amount of the drug to be administered is not particularly limited, and may be appropriately set according to the animal species, age, and the like. In the prevention and improvement method, the endurance improving method and the fatigue resistance method of the present invention, the administration method and the administration subject and the like are, for example, the same.

< food & drink >

The first food or drink of the present invention is a food or drink having a function of preventing or ameliorating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases, and autoimmune diseases, and is not limited at all except for containing the prophylactic/ameliorating agent of the present invention. The first food or drink of the present invention may further contain at least one of the endurance improver and the anti-fatigue agent of the present invention. The second food or drink of the present invention is not limited at all except for containing at least one of the endurance improving agent and the anti-fatigue agent of the present invention. The second food or drink of the present invention may further contain the prophylactic or ameliorating agent of the present invention. Hereinafter, the first food and drink and the second food and drink of the present invention may be collectively referred to as "food and drink". In the present invention, the food and drink includes general foods and health functional foods. The general food is not particularly limited, and examples thereof include processed grain food, processed vegetable food, processed fruit food, processed meat food, processed seafood, dairy products, beverages, and health foods. The food or drink of the present invention may further contain the above-mentioned prophylactic or ameliorating agent, endurance improver, and anti-fatigue agent as raw materials, additives, and the like. The cereal processed food is not particularly limited, and examples thereof include wheat flour, rice flour, cereal bars, batters, rice cakes, biscuits and the like. The processed vegetable food is not particularly limited, and examples thereof include vegetable paste, dried vegetables, and vegetable soup. The fruit processed food is not particularly limited, and examples thereof include fruit puree, dried fruit, and the like. The meat processed food is not particularly limited, and examples thereof include ham, bacon, sausage, and the like. The processed food of aquatic products is not particularly limited, and examples thereof include a marine food cooked in salt, a dried food, a fish sausage, a fish sweet potato cake, a fish cake, and a cylindrical fish cake. The dairy product is not particularly limited, and examples thereof include milk drinks, yogurt, ice cream, cheese, and the like. The beverage is not particularly limited, and examples thereof include cool beverages, green tea, black tea, and coffee. The functional health food is also generally called a functional food. Examples of the health functional food include specific health foods and nutritional functional foods.

The composition of the food or drink is not particularly limited, and examples thereof include various food materials, additives, stabilizers, and the like, in addition to the prevention and improvement agent, the endurance improver, and the anti-fatigue agent. The food or drink can be produced by a generally used formulation technique or the like. The type of animal to be used in the food or drink is not particularly limited, and examples thereof include humans, non-human mammals such as monkeys, cows, pigs, dogs, and cats, birds such as chickens, fish, and shellfish.

Examples

Next, examples of the present invention will be explained. However, the present invention is not limited to the following examples.

< example 1 >

In this example, the effect of red non-sulfur bacteria on inflammatory bowel disease was evaluated histologically using Dextran Sodium Sulfate (DSS) to induce colitis model mice.

(culture of cells)

First, 0.4 wt% of sodium acetate and 5 wt% of sucrose were added to a basal medium having a composition shown in table 4 below, and the pH was adjusted to 7.0 to prepare a medium for bacterial cells. The culture medium for the cells was added to a closed illumination culture tank so that the concentration of the cells was 1X 10% by volume⁶Cells/cm³The bacterial solution of Rhodobacter azotoformans BP0899 (accession No. NITE BP-644) was cultured at 30 ℃ for 7 days with stirring. After completion of the culture, the bacterial cells were collected from the obtained culture broth using a continuous centrifuge (Sharpless-Type) and the bacterial concentration was adjusted to 1X 10¹¹Cells/cm³And stored frozen.

TABLE 4

(preparation of cell powder)

The frozen cells were thawed naturally and dispensed into aspiration vials. The cells in the suction bottle were frozen at-45 ℃ for 20 minutes using a freeze dryer (manufactured by KOKAI Co., Ltd.). After freezing, the suction bottle was connected to a collector cooled to-45 ℃ and dried at room temperature (20-30 ℃) under 4-6 Pa to obtain freeze-dried cells. The lyophilized cells were pulverized with a propeller-type pulverizer (manufactured by Co., Ltd.) at a propeller rotation speed of 18000rpm to prepare a cell powder.

(test for administration in feed (a dose mixed in feed))

In the experiment of administering the mixture to the feed, 3 female mice of ICR line (Japan SLC, manufactured by Inc.) aged 5 weeks were used. For the purpose of inducing colitis in the aforementioned mice, DSS-containing water containing dextran sulfate sodium (molecular weight: 36000-50000, manufactured by ICN Biomedicals) at a concentration of 5w/v% was prepared. The powder of cells was added to a powder feed for mouse breeding (CF-2, manufactured by Japan clean) at a ratio of 0.1 wt% to prepare a feed containing cells. The mice were allowed to take the DSS-free water and the bacteria-containing feed for 1 week, and then the mice were allowed to take the DSS-containing water and the bacteria-containing feed for 7 days. On the 8 th day after the start of intake of the DSS-containing water, the colon of the mouse was harvested.

(histological evaluation)

The excised colon was fixed with 10v/v% formalin, and paraffin wax flakes were prepared by a conventional method. The slices were stained with hematoxylin, and then examined histologically using an optical microscope. Histological observation results such as inflammatory cell infiltration and mucosal epithelial swelling were quantified by the scores shown in the following table, and the average value was calculated.

(histological observation score)

< example 2 >

In this example, 5 ICR female mice (manufactured by Japan SLC) aged 5 weeks were administered water and feed in the same manner as in example 1, except that the intake period of the DSS-free water and the feed containing microbial cells was set to 4 weeks, and the ulcer formation and the regeneration epithelium were evaluated histologically.

< comparative example 1 >

In this example, 4 5-week-old female mice of ICR line (manufactured by Japan SLC) were fed with water and feed in the same manner as in example 1, and ulcer formation, inflammatory cell infiltration, swelling of mucosal epithelium, and histological evaluation of regenerated epithelium were carried out, except that a powdered feed for breeding mice (CF-2, manufactured by Japan clean) not containing the red non-sulfur bacteria was fed instead of the feed containing the cells.

< comparative example 2 >

In this example, 1 5-week-old female mice of the ICR line (manufactured by Japan SLC) were fed with water and feed in the same manner as in comparative example 1, and ulcer formation, inflammatory cell infiltration, swelling of mucosal epithelium, and regeneration of epithelium were evaluated histologically, except that DSS-free water was administered instead of DSS-containing water and colitis by DSS was not induced.

Table 5 below shows the average value of the scores of the histological observation results of inflammatory cell infiltration and mucosal epithelial swelling in example 1, comparative example 1, and comparative example 2. As shown in the table, in example 1, it was confirmed that the infiltration of inflammatory cells and the enlargement of mucosal epithelium were suppressed as compared with comparative example 1. In example 1, the feed containing the bacterial cells was ingested for 2 weeks, but no side effects were observed.

TABLE 5

Table 6 below shows the average of scores of histological observations of ulcer formation and regenerated epithelium in example 2, comparative example 1 and comparative example 2. As shown in the table, example 2 suppressed ulcer formation and promoted regeneration of the epithelium, compared to comparative example 1. This result is believed to be due to the rapid repair of the ulcer, which promotes epithelial regeneration. In example 2, the feed containing the bacterial cells was ingested for 5 weeks, but no side effects were observed.

TABLE 6

Fig. 1 to 3 show photographs of pathological tissues in example 2, comparative example 1, and comparative example 2. Fig. 1 (a) is a photograph of the pathological tissue of example 2 (magnification: 32 ×), and fig. 1 (B) is a photograph of the pathological tissue of example 2 (magnification: 80 ×). Fig. 2 (a) is a photograph of the pathological tissue of comparative example 1 (magnification: 32 ×), and fig. 2 (B) is a photograph of the pathological tissue of comparative example 1 (magnification: 80 ×). Fig. 3 (a) is a photograph of the pathological tissue of comparative example 2 (magnification: 32 ×), and fig. 3 (B) is a photograph of the pathological tissue of comparative example 2 (magnification: 80 ×).

As shown in FIG. 1, example 2, which ingested the feed containing the bacterial cells, suppressed ulcer formation and promoted regeneration of the epithelium. In the sheet shown in fig. 1, the histological score for ulcer formation was 1, and the histological score for regenerated epithelium was 4. On the other hand, as shown in FIG. 2, in comparative example 1 in which the feed containing the bacterial cells was not taken, ulcer formation was confirmed on the tissue surface (portion surrounded by an ellipse), and regeneration of the epithelium was hardly promoted. In the sheet shown in fig. 2, the histological score for ulcer formation was 4, and the histological score for regenerated epithelium was 1. In addition, as shown in FIG. 3, in comparative example 2 in which the feed containing the microbial cells and the water containing the DS were not taken, ulcer formation and regeneration of the epithelium were not confirmed, and the histological score of ulcer formation and regeneration of the epithelium was 0.

< example 3 >

In this example, mice model colitis were induced using Dextran Sodium Sulfate (DSS) and the effect of the amount of red non-sulfur bacteria administered on inflammatory bowel disease was evaluated histologically.

(preparation of cell powder)

In this example, bacterial cell powder was prepared in the same manner as in example 1.

(oral administration test)

For the oral administration test, 3 groups (2 to 4 groups) of 5-week-old female mice of ICR line (manufactured by Japan SLC) were used. For the purpose of inducing colitis in the aforementioned mice, DSS-containing water containing dextran sulfate sodium (molecular weight: 36000-50000, manufactured by ICN Biomedicals) at a concentration of 5w/v% was prepared. Each of the mice in the group was allowed to freely take the DSS-containing water, and the cell powder was administered 1 time for 1 day for 7 days at predetermined amounts (1, 6 and 10mg/kg, p.o.) as in example 1.

(histological evaluation)

The excised colon was fixed with 10v/v% formalin solution, and paraffin wax sheets were prepared by a conventional method, stained with hematoxylin, and then examined histologically using an optical microscope. The histological observation results of ulcer formation, inflammatory cell infiltration, swelling of mucosal epithelium, and hyperproliferation of regenerated epithelium and mucosa were quantified by the score in the same manner as in example 1, and the average value of each group was calculated.

< comparative example 3 >

In this example, 3 mice induced with DSS colitis were allowed to take the DSS-containing water and evaluated histologically for ulcer formation, inflammatory cell infiltration, and regenerated epithelium, in the same manner as in example 3, except that the bacterial cell powder was not administered orally.

Table 7 below shows the average of scores of histological observations of ulcer formation, inflammatory cell infiltration, and regenerated epithelium in example 3 and comparative example 3. In the table, the numbers in parentheses are the predetermined amounts (mg/kg, p.o.) of the cell powder administered in each group. As shown in the table, in example 3, ulcer formation was suppressed as compared with comparative example 3, and regenerative epithelialization was also suppressed in accordance with the suppression. In example 3, inhibition of infiltration of inflammatory cells was also confirmed in the group to which 6 or 10mg/kg, p.o. of the bacterial cell powder was administered every 1 day. In example 3, the bacterial cell powder was administered at 1 to 10mg/kg in p.o. for 7 days, but no side effects were observed.

TABLE 7

< example 4 >

In this example, a cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using a carbon Tetrachloride (THC) -induced liver injury model mouse as described below, and the effect of preventing liver injury was evaluated.

For the oral administration test, 3 groups (3 mice per group) of 5-week-old female mice of the ICR line (manufactured by Japan SLC) were used. The bacterial powder was administered to each group 1 time a day for 7 days at a dose of 10mg/kg, p.o.. Then, in order to induce liver injury, a predetermined amount (20, 100 and 500mg/kg, p.o.) of carbon tetrachloride (THC, manufactured by wako pure chemical industries, inc.) was administered to each of the groups 24 hours after the final administration. Then, blood was collected 24 hours after the THC administration, and GOT activity (Karmen unit) in the blood was measured to calculate the average GOT value of each group. Further, as a control, GOT activity in blood was measured in the same manner as in the above-described groups except that the cell powder and the THC were not orally administered, and a GOT average value was calculated. Then, the relative activity value of GOT for each group was calculated by the following formula (1).

GOT relative activity (%) - (average GOT for each group)/(average GOT for control) × 100 · (1)

< comparative example 4 >

In this example, an oral administration test was carried out in the same manner as in example 4 except that the bacterial cell powder was not orally administered.

Table 8 below shows the relative activity (%) of GOT classified by THC administration amount (mg/kg, p.o.) in example 4 and comparative example 4. In the table, the numbers in parentheses are the average values of the GOT measurement values in the blood. As shown in the table, the relative activity value of GOT at 20mg/kg p.o.thc was 100.3% in example 4, and 200.0% in comparative example 4. The relative activity of GOT at 100mg/kg p.o.thc was 216.2% in example 4, and 435.4% in comparative example 4. Further, the relative activity value of GOT at 500mg/kg p.o.THC was 311.3% in example 4, and 572.4% in comparative example 4. That is, in example 4, the GOT activity was suppressed about 1/2 times as compared with comparative example 4, and the effect of suppressing liver damage was confirmed. In example 4, the bacterial cell powder was administered at 10mg/kg in p.o. for 7 days, but no side effects were observed.

TABLE 8

< example 5 >

In this example, a cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using a carbon Tetrachloride (THC) -induced liver injury model mouse as described below, and the effect of preventing liver injury was evaluated.

For the oral administration test, 3 groups (3 mice per group) of 5-week-old female mice of the ICR line (manufactured by Japan SLC) were used. A predetermined amount (3, 6 and 10mg/kg, p.o.) of the microbial cell powder was administered to each group 1 time per day and 7 days per day. Then, to induce liver injury, 100mg/kg of carbon tetrachloride (THC, manufactured by wako pure chemical industries, Ltd.) was administered at a p.o. 24 hours after the final administration. Then, blood was collected 24 hours after the administration of the THC, and GOT activity in the blood was measured (Karmen unit), and the average GOT value of each group was calculated. In addition, as a control, GOT activity in blood was measured in the same manner as in the case of the above-mentioned cell powder group, except that the cell powder and THC were not orally administered, and a GOT average value was calculated. Then, the relative activity of GOT in each of the above groups was calculated in the same manner as in example 4.

< comparative example 5 >

In this example, an oral administration test was carried out in the same manner as in example 5 except that 3 ICR-line female mice (manufactured by Japan SLC) aged 5 weeks were used, and the cell powder was not orally administered, to calculate the relative GOT activity.

Table 9 below shows the relative activity (%) of GOT classified by the amount of bacterial powder (mg/kg, p.o.) used in example 5 and the relative activity (%) of GOT of comparative example 5. In the table, the numbers in parentheses are the average values of the GOT measurement values in the blood. As shown in the table, the GOT activity was suppressed and the effect of suppressing the liver damage was confirmed in each group of example 5 to which 3 to 10mg/kg of the p.o. cell powder was administered, as compared with comparative example 5. In example 5, the cell powder was administered at 3 to 10mg/kg in p.o. for 7 days, but no side effects were observed.

TABLE 9

< example 6 >

In this example, a cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using a carbon Tetrachloride (THC) -induced liver injury model mouse as described below, and the effect thereof was evaluated histologically.

(oral administration test using liver injury-inducing model mouse)

In this example, an oral administration test was carried out in the same manner as in example 4 except that 10mg/kg of the cell powder was administered 1 time per 1 day for 1 day or 7 days, and 100mg/kg of p.o. carbon tetrachloride (THC, manufactured by Wako pure chemical industries, Ltd.) was administered 24 hours after the final administration. In addition, another group (3 individuals) to which the cell powder and the THC were not orally administered was used as a control.

(histological evaluation)

After 24 hours of administration of the THC, the liver was extirpated. Then, the excised liver was fixed with 10v/v% formalin solution, and paraffin slices were prepared by a conventional method, and after hematoxylin staining, histological observation was performed using an optical microscope. The histological observation results of hepatic lobular necrosis of liver and inflammatory cell infiltration were quantified by using scores in the same manner as in example 1, and the average value of each group and control was calculated.

< comparative example 6 >

In this example, an oral administration test was carried out in the same manner as in example 6 except that the bacterial cell powder was not orally administered, and histological evaluation was carried out for hepatic necrosis and inflammatory cell infiltration in lobules of the liver.

Table 10 below shows the average of the scores of the histological observation results of hepatic cell necrosis and inflammatory cell infiltration at the center of the lobule of the aforementioned liver in example 6 and comparative example 6. In the table, the days in parentheses are the days of administration of the bacterial cell powder. As shown in the table, in example 6, hepatic cell necrosis at the center of lobules and infiltration of inflammatory cells were suppressed as compared with comparative example 6. That is, THC-induced liver injury is suppressed by oral administration of the above-described bacterial cell powder. In example 6, the bacterial cell powder was administered at 10 mg/kg/p.o. for 1 to 7 days, but no side effects were observed.

Watch 10

< example 7 >

In this example, a bacterial cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using an ovalbumin-sensitized allergy model mouse as described below, and the effect of the Th1/Th2 balance on ovalbumin-sensitized allergy was evaluated.

(IgE assay in serum)

First, 2mg of ovalbumin (grade V, manufactured by Sigma Co.) was dissolved in 0.2mL of physiological saline, and then 2mg of aluminum hydroxide gel (No.019-19501, LotWKJ 4431, manufactured by Wako pure chemical industries, Ltd.) was added and suspended to prepare an ovalbumin/aluminum hydroxide gel. In the oral administration test, 5 female mice of BALB/c line (manufactured by Japan SLC) aged 4 weeks were used. 10mg/kg of the bacterial powder was administered to each of the mice 1 time a day for 2 weeks at p.o. Then, the ovalbumin/aluminum hydroxide gel was administered intraperitoneally on the final day of administration of the cell powder. Further, additional immunization was performed 10 days after the final administration day, and blood was collected 1 week after the additional immunization. Serum was collected from the collected blood, and the amount of IgE in the serum was measured according to the instructions of ELISA Kit (Bethy Laboratories, Inc.) to calculate an average of 5.

(measurement of IL-4 amount in serum)

In this measurement, the cell powder was administered in the same manner as in the measurement of the amount of IgE in serum, and ovalbumin/aluminum hydroxide gel was prepared and administered to collect blood, except that 2 BALB/c-line female mice (manufactured by Japan SLC) aged 4 weeks were used. Then, serum was collected from the collected blood, and the amount of IL-4 in the serum was measured according to the instruction manual of IL-4, mouse, ELISAKit (96well, Bethy Laboratories), to calculate the average value of 2 individuals.

(measurement of cytokine amount in culture supernatant of spleen cells)

In this measurement, the cell powder was administered in the same manner as in the measurement of the amount of IgE in serum, and ovalbumin/aluminum hydroxide gel was prepared and administered, except that 3 BALB/c-line female mice (manufactured by Japan SLC) aged 4 weeks were used. Then, spleens were harvested 1 week after the booster immunization. Spleen cells were collected from the spleen and cultured in RPMI 1640 medium (Gibco laboratory) supplemented with 100. mu.g/mL ovalbuminProducts of ies corporation) of 5X 10⁶The cells/mL density suspension, in air 95%, carbon dioxide concentration 5%, 37 ℃ under the condition of 3 days. After completion of the culture, the culture supernatant was collected, and the amounts of IFN-. gamma.IL-2, IL-4 and IL-5 in the supernatant were measured according to the instructions of ELISA kits (manufactured by Bethy Laboratories) shown in Table 11 below, and the average of 3 samples was calculated.

TABLE 11

< comparative example 7 >

In this example, an oral administration test was carried out in the same manner as in example 7 except that the above-mentioned bacterial cell powder was not orally administered, and the amounts of IgE and IL-4 in serum were measured, and the amounts of IFN-. gamma., IL-2, IL-4 and IL-5 in the culture supernatant of spleen cells were measured to calculate average values.

The average values of the IgE amounts in the above-mentioned sera in example 7 and comparative example 7 are shown in table 12 below. As shown in the table, example 7 in which the bacterial cell powder was administered showed a decrease in the amount of IgE as compared with comparative example 7 in which the bacterial cell powder was not administered.

TABLE 12

The following Table 13 shows the results of measurement of the IL-4 amount in the serum described above in example 7 and comparative example 7. As shown in the table, the amount of IL-4 was reduced in example 7 in which the cell powder was administered, compared with comparative example 7 in which no cell powder was administered. That is, since the amount of IL-4, which is a Th2 type cytokine, is suppressed and the amount of IgE is suppressed as described above, it is found that the above-mentioned cell powder can improve the imbalance of Th1/Th2, which causes allergic reaction, by suppressing the secretion of Th2 type cytokine.

Watch 13

The results of measuring the amounts of IFN-. gamma.IL-2, IL-4 and IL-5 in the aforementioned culture supernatants in example 7 and comparative example 7 are shown in Table 14 below. As shown in the table, in example 7 in which the cell powder was administered, the amounts of Th1 type cytokines IFN-. gamma.and IL-2 were increased and the amounts of Th2 type cytokines IL-4 and IL-5 were decreased, compared with comparative example 7 in which no cell powder was administered. That is, it is known that the allergic reaction is caused by the predominance of Th2, and the cell powder suppresses the secretion of Th2 type cytokines, thereby improving the imbalance of Th1/Th2 and exhibiting an antiallergic effect. In example 7, 10mg/kg of the cell powder was administered 1 time a day for 2 weeks, but no side effects were observed.

TABLE 14

< example 8 >

In this example, a bacterial cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using a mouse in which an intradermal inflammation (edema) caused by ovalbumin was caused, as described below, to evaluate an antiallergic effect.

First, an ovalbumin/aluminum hydroxide gel was prepared in the same manner as in example 7, except that the concentration of ovalbumin (grade V, manufactured by sigma) was changed to 20. mu.g. The bacterial cell powder and the ovalbumin/aluminum hydroxide gel were administered to each mouse in the same manner as in example 7, except that 6 female mice of BALB/c line (manufactured by Japan SLC) aged 4 weeks were used. After 10 days from the final administration day of the cell powder, 20. mu.L of the ovalbumin/aluminum hydroxide gel was inoculated onto the sole of each mouse. After 24 hours of inoculation, the Volume of edema produced in the sole was measured using a Volume Meter (Volume Meter) MK-550 (manufactured by street mechanical corporation), and the average value was calculated.

< comparative example 8-1 >

In this example, an oral administration test was carried out in the same manner as in example 8 except that 3 mice were used and neither the cell powder nor the ovalbumin/aluminum hydroxide gel was administered, and the volume of edema formed in the sole of the foot was measured and the average value was calculated.

< comparative example 8-2 >

In this example, an oral administration test was carried out in the same manner as in example 8 except that the cell powder was not orally administered as the non-administered cell group, and the volume of edema produced in the sole of the foot was measured to calculate an average value.

The measurement results of the aforementioned edema volume in example 8, comparative example 8-1 and comparative example 8-2 are shown in the table of FIG. 4. In the table of the figure, the vertical axis represents the average volume (mL) of the edema. As shown in the figure, the edema volume of each example was 0.35mL in comparative example 8-1 (untreated group), 0.95mL in comparative example 8-2 (non-cell-administered group), and 0.63mL in example 8. Therefore, in example 8 in which the cell powder was administered, the edema volume was suppressed as compared with comparative example 8-2 in which no cell powder was administered. That is, the cell powder exhibits an antiallergic effect against an allergic reaction caused by intradermal immunization. In example 8, 10mg/kg of the cell powder was administered 1 time a day for 2 weeks, but no side effects were observed.

< example 9 >

In this example, a bacterial cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using an adjuvant arthritis rat as a rheumatoid arthritis model as described below, and the effect on rheumatoid arthritis (autoimmune disease) was evaluated.

First, 0.6mg of Mycobacterium tuberculosis (M.butulicum, Lot 0640-33, manufactured by Difco) was suspended in 0.1mL of liquid paraffin (manufactured by Bao chemical Co.) to prepare a complete adjuvant. In the oral administration test, 5 Wistar male rats (150-160 g, manufactured by Japan SLC) aged 5 weeks were used. The hind limb soles of each rat were inoculated with 0.1mL of the aforementioned adjuvant. The cell powder was administered to each rat at a dose of 20mg/kg p.o. 1 time for 1 day and 21 days from the day of inoculation. Then, the volume of edema produced in the sole was measured using a volumeter MK-550 (manufactured by komachi city co.) on days 3, 17, and 21 from the inoculation day, and the average value was calculated. On the same day, photographs were taken of tarsal bones, metatarsal bones and phalanges of hind limbs using a soft X-ray apparatus, and the morphology of bone disease deformation caused by adjuvant arthritis was observed.

< comparative example 9-1 >

In this example, an oral administration test was performed in the same manner as in example 9 except that the cell powder was not administered as the untreated group, and the average value of the edema volume was measured to perform morphological observation.

< comparative example 9-2 >

In this example, as a positive control group, an oral administration test was carried out in the same manner as in example 9 except that indomethacin (manufactured by sigma) as an anti-inflammatory agent was administered at 0.1mg/kg, p.o. in place of the above-mentioned bacterial cell powder, and the edema volume was measured to calculate an average value, and morphological observation was carried out.

The results of measuring the edema volumes described above in example 9, comparative example 9-1 and comparative example 9-2 are shown in the table of FIG. 5. In the table of the figure, the vertical axis represents the average value (mL) of edema volume, and the horizontal axis represents the number of days (days) after inoculation. In addition, the respective histograms show the results of comparative example 9-1, example 9 and comparative example 9-2 in this order from left to right. As shown in the figure, on days 17 and 21 after inoculation, the edema volume of example 9 (the group to which the bacterial cell powder was administered) was suppressed as compared with comparative example 9-1 (the untreated group) in the same manner as in comparative example 8-2 (the positive control group). That is, in example 9 in which the cell powder was administered, compared with comparative example 9-1 in which no cell powder was administered, the formation of edema due to adjuvant arthritis was suppressed in the same manner as in comparative example 9-2 in which indomethacin was administered.

Fig. 6 is a photograph showing bone disease distortion of hind limbs in the foregoing examples. FIG. 6(A) is a photograph of example 9, FIG. 6(B) is a photograph of comparative example 9-1, and FIG. 6(C) is a photograph of comparative example 9-2. As shown in fig. 6(B), bone lesions (arrowheads) due to adjuvant arthritis were observed in comparative example 9-1 (untreated group). On the other hand, as shown in fig. 6(a) and (C), inflammation and bone lesions due to adjuvant arthritis were suppressed in example 9 in which the bacterial cell powder was administered, as in comparative example 9-2 in which indomethacin was administered. Thus, the bacterial cell powder of the present invention is considered to be useful for preventing and improving autoimmune diseases such as chronic rheumatoid arthritis. In example 9, 20mg/kg of the microbial cell powder was administered 1 time a day for 21 days, but no side effects were observed.

< example 10 >

In this example, a bacterial cell powder prepared in the same manner as in example 1 was subjected to an oral administration test using an ovalbumin-sensitized anaphylactic reaction model mouse as described below, and the effect of histamine release in an ovalbumin-sensitized anaphylactic reaction was evaluated.

The bacterial cell powder was administered in the same manner as in the measurement of the amount of IgE in serum described in example 7 except that 19 BALB/c-line female mice (manufactured by Japan SLC) aged 4 weeks were used, and the ovalbumin/aluminum hydroxide gel was prepared and administered to collect blood. Then, serum was collected from the collected blood, and the amount of histamine in the serum was measured according to the instruction manual of histamine ELISA Kit (manufactured by immunoltech) to calculate an average value.

< comparative example 10-1 >

In this example, an oral administration test was carried out in the same manner as in example 10 except that 15 mice were used and the bacterial cell powder was not orally administered, and the amount of histamine in serum was measured to calculate an average value.

< comparative example 10-2 >

In this example, an oral administration test was carried out in the same manner as in example 10 except that 13 mice were used and the ovalbumin/aluminum hydroxide gel and the cell powder were not orally administered, and the amount of histamine in serum was measured to calculate an average value.

FIG. 7 and Table 15 below show the results of the average values of the amounts of histamine in the above-mentioned sera in example 10, comparative example 10-1 and comparative example 10-2. As shown in FIG. 7 and Table 15 below, example 10 to which the bacterial cell powder was administered showed a decrease in the amount of histamine as compared with comparative example 10-1 to which no bacterial cell powder was administered. Thus, it was confirmed that the administration of the bacterial powder suppressed the release of histamine.

Watch 15

< example 11 >

In this example, the improvement of endurance and the anti-fatigue effect of the bacterial powder prepared in the same manner as in example 1 were evaluated.

A4-week-old ddY male mouse (manufactured by Japan SLC) was subjected to the drape method as described below, and a mouse with a drape duration of 40 to 60 seconds was taken out. The overhang method is performed as follows. First, the mouse was suspended on a horizontally disposed suspension stick with the hind paw being weighted by 10% of the body weight, and the suspension duration (seconds) was measured.

The bacterial cell powder was dissolved in physiological saline for injection so that the concentration thereof became 1mg/mL to prepare a sample. The sample was orally administered to 10 mice taken out in an amount of 0.1mL per 10g of mouse body weight for 1 day and 10 days so that the amount of administration became 10mg/kg, p.o./day.

After 11 days of sample administration, the duration of drape was determined by the foregoing drape method. This measurement was defined as the 1 st measurement. Further, after 30 minutes from the 1 st measurement, the drape duration was measured again by the drape method. This measurement was designated as the 2 nd measurement.

< comparative example 11 >

In this example, oral administration was carried out in the same manner as in example 11 except that the bacterial cell powder was not orally administered, and the suspension duration was measured by the suspension method.

Fig. 8 shows a table of the measurement results of the drape duration in example 11 and comparative example 11. In the table of fig. 8, the vertical axis represents the drape duration (seconds), and the horizontal axis represents the measurement result at the 1 st time and the measurement result at the 2 nd time in this order from left to right. In the table of fig. 8, white columns represent the measurement results of comparative example 11 (group N), and black columns represent the measurement results of example 11 (group T).

As shown in fig. 8, the duration of the 1 st drape of example 11(T group) administered with the 10-day sample was 39.2 seconds, and the duration of the 1 st drape of comparative example 11(N group) was 31.4 seconds. That is, in the 1 st measurement, the drape duration of example 11 was extended 1.2 times as compared with comparative example 11. In the 2 nd measurement, the drape duration of example 11(T group) was 36.8 seconds, and that of comparative example 11(N group) was 17.2 seconds. That is, in the 2 nd measurement, the drape duration of example 11 was extended 2.1 times as compared with comparative example 11, and the difference between the two was significant (P ═ 0.0034). Thus, the sample containing the bacterial cell powder was orally administered for 10 days, whereby the endurance was improved and the anti-fatigue effect was exhibited.

< example 12 >

In this example, oral administration was carried out in the same manner as in example 11 except that the administration period of the sample containing the bacterial cell powder was set to 20 days, and the suspension duration was measured by the suspension method.

< comparative example 12 >

In this example, oral administration was carried out in the same manner as in example 12 except that the bacterial cell powder was not orally administered, and the suspension duration was measured by the suspension method.

Fig. 9 is a table showing the measurement results of the drape duration in example 12 and comparative example 12. In the table of fig. 9, the vertical axis represents the drape duration (seconds), and the horizontal axis represents the measurement result at the 1 st time and the measurement result at the 2 nd time in this order from left to right. In the table of fig. 9, white columns represent the measurement results of comparative example 12(N group), and black columns represent the measurement results of example 12(T group).

As shown in fig. 9, the duration of the 1 st drape of example 12(T group) administered with the 20-day sample was 68.2 seconds, and the duration of the 1 st drape of comparative example 12(N group) was 54.3 seconds. That is, in the 1 st measurement, the drape duration of example 12 was extended 1.2 times as compared with comparative example 12. In the 2 nd measurement, the drape duration was 57.1 seconds for example 12(T group) and 26.8 seconds for comparative example 12(N group). That is, in the 2 nd measurement, the drape duration of example 12 was extended 2.1 times as compared with comparative example 12, and the difference between the two was significant (P ═ 0.0001). Thus, the sample containing the bacterial cell powder was orally administered for 20 days, whereby the endurance was improved and the anti-fatigue effect was exhibited.

It is clear from examples 1 to 10 and comparative examples 1 to 10 that the ingestion of the red non-sulfur bacterium of the present invention can inhibit ulcer formation, inflammatory cell infiltration, and swelling of mucosal epithelium in the large intestine, and further can promote epithelial regeneration, and prevent and improve inflammatory diseases. By taking the red non-sulfur bacterium of the present invention, hepatic necrosis at the center of lobule in the liver and infiltration of inflammatory cells can be suppressed, and inflammatory diseases can be prevented. By taking the red non-sulfur bacterium of the present invention, the amount of IgE and histamine in serum is reduced, and secretion of Th2 type cytokines can be suppressed, thereby improving the imbalance of Th1/Th2 and suppressing allergy. By ingesting the red non-sulfur bacterium of the present invention, it is possible to suppress allergic reactions caused by intradermal immunity and, in addition, to prevent and improve autoimmune diseases. Further, it is clear from examples 11 and 12 and comparative examples 11 and 12 that the intake of the red non-sulfur bacteria of the present invention can improve endurance and suppress fatigue. The red non-sulfur bacterium of the present invention has no side effects due to long-term ingestion, and exhibits high safety.

Industrial applicability

As described above, the red non-sulfur bacterium of the present invention has various effects as follows: various effects such as inhibition of ulcer formation, inhibition of inflammatory cell infiltration, inhibition of mucosal epithelial swelling, epithelial regeneration, inhibition of hepatocyte necrosis and promotion of hepatocyte swelling, reduction of IgE amount and histamine amount in serum, inhibition of Th2 type cytokine secretion, improvement of Th1/Th2 imbalance, inhibition of anaphylaxis, inhibition of autoimmune disease, improvement of endurance, anti-fatigue, etc.; can prevent and improve at least one disease selected from the group consisting of inflammatory diseases, allergic diseases and autoimmune diseases, improve endurance, and inhibit fatigue. The red non-sulfur bacterium of the present invention is highly safe, can be administered for the purpose of prevention before the disease occurs, and can be administered for a long period of time. Therefore, according to the present invention, there can be provided a prophylactic/ameliorating agent, a persistent activity-enhancing agent, an anti-fatigue agent, a pharmaceutical product, a food or drink, which is useful for preventing and ameliorating at least one disease selected from the group consisting of an inflammatory disease, an allergic disease, and an autoimmune disease, improving the persistent activity, and relieving fatigue, and has high safety, and the application range thereof is wide and is not limited.

Claims (17)

1. A prophylactic/ameliorating agent for at least one disease selected from the group consisting of inflammatory diseases, allergic diseases and autoimmune diseases,

containing at least one of a red non-sulfur bacterium and a culture of the red non-sulfur bacterium,

the red non-sulfur bacteria contain nitrogen-fixing red bacteria (Rhodobacter azotoformans) which are the Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain with the deposition number of NITEBP-644.

2. The prophylactic/ameliorating agent according to claim 1, wherein the Rhodobacter azotoformans (Rhodobacter azotoformans) has the following bacteriological characteristics (1) to (30):

(1) shape of the cells: rod-like or ovoid shape

(2) Polymorphism: is free of

(3) Size of the cells: 0.8 μm × 1.0 μm

(4) Presence or absence of motility: is provided with

(5) Presence or absence of spores: is free of

(6) Gloss on regular agar culture: is provided with

(7) Pigment production on normal agar culture: is provided with

(8) Presence or absence of surface development in ordinary broth culture: is free of

(9) Presence or absence of turbidity of the medium in the culture in ordinary broth: is provided with

(10) Liquefaction of gelatin during gelatin puncture culture: negative of

(11) Solidification of litmus milk during culture: is free of

(12) Liquefaction of litmus milk during culture: is free of

(13) Gram staining property: negative of

(14) Reduction of nitrate: is free of

(15) And (3) denitrification reaction: without or with

(16) And (3) MR test: negative of

(17) Indole production: is free of

(18) Generation of hydrogen sulfide: is free of

(19) Hydrolysis of starch: is free of

(20) Utilization of citric acid, kreb: is free of

(21) Utilization of inorganic nitrogen source, ammonium salt: is provided with

(22) Production of Catalase: positive for

(23) Production of oxidase: positive for

(24) Anaerobic fertility: is provided with

(25) O-F test: oxidation, negativity/fermentation, negativity

(26) β -galactosidase activity: negative of

(27) Arginine double hydrolase activity: negative of

(28) Lysine decarboxylase activity: negative of

(29) Tryptophan deaminase activity: negative of

(30) Gelatinase activity: and (4) negativity.

3. The prophylactic/ameliorating agent according to claim 1, wherein the 16S rRNA of the Rhodobacter azotoformans (Rhodobacter azotoformans) has a base sequence represented by SEQ ID NO. 1.

4. A endurance improving agent comprising at least one of a red non-sulfur bacterium and a culture of the red non-sulfur bacterium,

the red non-sulfur bacteria contain nitrogen-fixing red bacteria (Rhodobacter azotoformans) which are the Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain with the deposition number of NITEBP-644.

5. The endurance improver according to claim 4, wherein the Rhodobacter azotoformans (Rhodobacter azotoformans) has the following bacteriological characteristics (1) to (30):

(1) shape of the cells: rod-like or ovoid shape

(2) Polymorphism: is free of

(3) Size of the cells: 0.8 μm × 1.0 μm

(4) Presence or absence of motility: is provided with

(5) Presence or absence of spores: is free of

(6) Gloss on regular agar culture: is provided with

(7) Pigment production on normal agar culture: is provided with

(8) Presence or absence of surface development in ordinary broth culture: is free of

(9) Presence or absence of turbidity of the medium in the case of ordinary broth culture: is provided with

(10) Liquefaction of gelatin during gelatin puncture culture: negative of

(11) Solidification of litmus milk during culture: is free of

(12) Liquefaction of litmus milk during culture: is free of

(13) Gram staining property: negative of

(14) Reduction of nitrate: is free of

(15) And (3) denitrification reaction: without or with

(16) And (3) MR test: negative of

(17) Indole production: is free of

(18) Generation of hydrogen sulfide: is free of

(19) Hydrolysis of starch: is free of

(20) Utilization of citric acid, kreb: is free of

(21) Utilization of inorganic nitrogen source, ammonium salt: is provided with

(22) Production of Catalase: positive for

(23) Production of oxidase: positive for

(24) Anaerobic fertility: is provided with

(25) O-F test: oxidation, negativity/fermentation, negativity

(26) β -galactosidase activity: negative of

(27) Arginine double hydrolase activity: negative of

(28) Lysine decarboxylase activity: negative of

(29) Tryptophan deaminase activity: negative of

(30) Gelatinase activity: and (4) negativity.

6. The endurance improver according to claim 4, wherein the 16S rRNA of the Rhodobacterium azotoformans (Rhodobacter azotoformans) has a base sequence represented by SEQ ID NO. 1.

7. An anti-fatigue agent characterized by comprising at least one of a red non-sulfur bacterium and a culture of the red non-sulfur bacterium,

the red non-sulfur bacteria contain nitrogen-fixing red bacteria (Rhodobacter azotoformans) which are the Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain with the deposition number of NITEBP-644.

8. The anti-fatigue agent according to claim 7, wherein the Rhodobacter azotoformans (Rhodobacter azotoformans) has the following bacteriological characteristics (1) to (30):

(1) shape of the cells: rod-like or ovoid shape

(2) Polymorphism: is free of

(3) Size of the cells: 0.8 μm × 1.0 μm

(4) Presence or absence of motility: is provided with

(5) Presence or absence of spores: is free of

(6) Gloss on regular agar culture: is provided with

(7) Pigment production on normal agar culture: is provided with

(8) Presence or absence of surface development in ordinary broth culture: is free of

(9) Presence or absence of turbidity of the medium in the case of ordinary broth culture: is provided with

(10) Liquefaction of gelatin during gelatin puncture culture: negative of

(11) Solidification of litmus milk during culture: is free of

(12) Liquefaction of litmus milk during culture: is free of

(13) Gram staining property: negative of

(14) Reduction of nitrate: is free of

(15) And (3) denitrification reaction: without or with

(16) And (3) MR test: negative of

(17) Indole production: is free of

(18) Generation of hydrogen sulfide: is free of

(19) Hydrolysis of starch: is free of

(20) Utilization of citric acid, kreb: is free of

(21) Utilization of inorganic nitrogen source, ammonium salt: is provided with

(22) Production of Catalase: positive for

(23) Production of oxidase: positive for

(24) Anaerobic fertility: is provided with

(25) O-F test: oxidation, negativity/fermentation, negativity

(26) β -galactosidase activity: negative of

(27) Arginine double hydrolase activity: negative of

(28) Lysine decarboxylase activity: negative of

(29) Tryptophan deaminase activity: negative of

(30) Gelatinase activity: and (4) negativity.

9. The anti-fatigue agent according to claim 7, wherein the 16S rRNA of Rhodobacter azotoformans (Rhodobacter azotoformans) has a base sequence represented by SEQ ID NO. 1.

10. A pharmaceutical product for preventing or treating at least one disease selected from the group consisting of inflammatory diseases, allergic diseases and autoimmune diseases, which comprises the prophylactic/ameliorating agent according to claim 1.

11. A pharmaceutical product containing at least one of the endurance improving agent of claim 4 and the anti-fatigue agent of claim 7.

12. A food or drink having a function of preventing or improving at least one disease selected from the group consisting of inflammatory diseases, allergic diseases and autoimmune diseases, which contains the prophylactic/ameliorating agent according to claim 1.

13. A food or drink comprising at least one of the endurance improver according to claim 4 and the anti-fatigue agent according to claim 7.

14. A red non-sulfur bacterium, which is Rhodobacterium azotoformans (Rhodobacter azotoformans) BP0899 strain deposited under the accession number NITE BP-644.

15. Use of a red non-sulfur bacterium for the preparation of a prophylactic or ameliorating agent for a method for the prophylactic or ameliorating of at least one disease selected from the group consisting of inflammatory diseases, allergic diseases and autoimmune diseases,

the method for preventing or ameliorating a red non-sulfur bacterium comprises a step of administering a preventive or ameliorating agent containing at least one of the red non-sulfur bacterium and a culture of the red non-sulfur bacterium,

the red non-sulfur bacteria contain nitrogen-fixing red bacteria (Rhodobacter azotoformans) which are the Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain with the deposition number of NITEBP-644.

16. A use of red non-sulfur bacteria for producing a stamina-improving agent for use in a stamina-improving method comprising a step of administering a stamina-improving agent comprising at least one of red non-sulfur bacteria and a culture of the red non-sulfur bacteria,

the red non-sulfur bacteria contain nitrogen-fixing red bacteria (Rhodobacter azotoformans) which are the Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain with the deposition number of NITEBP-644.

17. Use of a red non-sulfur bacterium for the preparation of an anti-fatigue agent for use in an anti-fatigue method comprising a step of administering an anti-fatigue agent comprising at least one of a red non-sulfur bacterium and a culture of the red non-sulfur bacterium,

the red non-sulfur bacteria contain nitrogen-fixing red bacteria (Rhodobacter azotoformans) which are the Rhodobacter azotoformans (Rhodobacter azotoformans) BP0899 strain with the deposition number of NITEBP-644.