Akkermansia muciniphila VB202 and use thereof

Abstract

Provided are Akkermansia muciniphila VB202 and a use thereof. The strain was deposited at the China General Microbiological Culture Collection Center on 29 August 2023, with the accession number being CGMCC No. 28295. The provided Akkermansia muciniphila has relatively strong antibacterial ability, and can thus be used as an alternative to antibiotics; additionally, the strain has good hypoglycemic ability and intestinal tolerance, and can thus be used for preparing blood glucose control products.

Description

Akkermansia muciniphila VB202 and its application

[0001] Priority information

[0002] This application claims priority to the Chinese patent application with application number 202311658889.8 filed with the China Patent Office on December 5, 2023, entitled “A Muciniphilic Akkermansia VB202 and Its Applications,” the entire contents of which are incorporated by reference into this application. Technical Field

[0003] The present invention relates to the technical field of microorganisms, and in particular to Akkermansia muciniphila VB202 and applications thereof. Background Art

[0004] Akkermansia muciniphila (AKK) is a novel intestinal microorganism first discovered in healthy human feces by Derrien et al. in 2004. It belongs to the genus Akkermansia in the family Verrucomicrobiaceae of the phylum Verrucomicrobia. It colonizes the intestinal mucosa using mucin as its sole nitrogen and carbon source. Akkermansia muciniphila is widely distributed in the intestines of healthy infants and adults, stably colonizing the intestine by the first year of life and constituting 1% to 3% of the total intestinal microbiome in adults, making it a crucial component of the intestinal microbiota. Studies have found that Akkermansia muciniphila is significantly reduced in the intestines of patients with various diseases, and its abundance correlates with disease status. Therefore, it holds great promise for development as a next-generation probiotic, following Bifidobacterium and Lactobacillus (O'Toole PW, Marchesi JR, Hill C. Next-generation probiotics: the spec-trum from probiotics to live biotherapeutics [J]. Nat Microbiol, 2017, 2:17057).

[0005] Different types of pathogens not only pose food safety risks but can also infect humans and animals, causing illnesses and even life-threatening conditions. Currently, antibiotics or synthetic chemicals are primarily used to effectively control or eliminate pathogens. However, increasing antibiotic resistance among pathogens, the frequent occurrence of irreversible damage from chemical residues and environmental pollution, pose serious challenges to human health and sustainable socioeconomic development. Announcement No. 194 issued by my country's Ministry of Agriculture and Rural Affairs in July 2019 stated that, effective January 1, 2020, the addition of antibiotics to feed will be completely banned in my country, necessitating the urgent need for research on alternatives. Probiotics produce metabolites that inhibit pathogens during their growth and reproduction. They can also improve the acidic environment of the animal's intestinal tract, optimize the homeostasis of intestinal microorganisms, stimulate intestinal mucosal immunity, inhibit the colonization of harmful bacteria, and enhance immunity, thereby improving animal performance and health. Natural pathogen control holds great promise.

[0006] Currently, there are many studies on the correlation between AKK content and various disease conditions, but there are fewer studies on its antibacterial properties. Therefore, there is an urgent need to provide an AKK bacterium with antibacterial properties.

[0007] Summary of the Invention

[0008] The present invention aims to solve one of the technical problems in the related art at least to a certain extent.

[0009] To this end, the first aspect of the present invention provides a microorganism, which is Akkermansia muciniphila VB202, deposited in the General Microbiology Center of the China Culture Collection Administration on August 29, 2023, with a deposit number of CGMCC No. 28295.

[0010] The Akkermansia muciniphila VB202 provided by the present invention has good antibacterial ability, intestinal tolerance and blood sugar lowering ability. On the one hand, it can inhibit the growth of harmful bacteria such as Helicobacter pylori, Listeria monocytogenes and Shigella, helping the body to restore the balance of the microbiome without causing the development of drug resistance. It can be used as a safer alternative to antibiotics. On the other hand, this strain has a blood sugar lowering effect and can be used to prepare blood sugar control products.

[0011] According to an embodiment of the present invention, the microorganism has a 16S rDNA sequence as shown in SEQ ID NO: 1.

[0012] The second aspect of the present invention provides a fermentation broth, which is fermented by the microorganism described in the first aspect.

[0013] A third aspect of the present invention provides a bacterial suspension comprising the microorganisms described in the first aspect.

[0014] The fourth aspect of the present invention provides use of the microorganism described in the first aspect, the fermentation broth described in the second aspect, or the bacterial suspension described in the third aspect in the preparation of medicines, feeds, and additives for inhibiting the activity of pathogenic bacteria.

[0015] According to an embodiment of the present invention, the pathogenic bacteria is selected from at least one of Helicobacter pylori, Listeria monocytogenes, and Shigella.

[0016] The fifth aspect of the present invention provides use of the microorganism described in the first aspect, the fermentation broth described in the second aspect, or the bacterial suspension described in the third aspect in the preparation of drugs, feeds, and additives for controlling or lowering blood sugar.

[0017] The sixth aspect of the present invention provides a composition comprising at least one of the microorganisms described in the first aspect, the fermentation broth described in the second aspect, and the bacterial suspension described in the third aspect.

[0018] According to an embodiment of the present invention, the composition further comprises an excipient and / or a carrier.

[0019] According to an embodiment of the present invention, the excipient includes at least one selected from a binder, a disintegrant, a lubricant, a glidant, a stabilizer, a filler, a diluent, and a sustained-release agent.

[0020] According to an embodiment of the present invention, the carrier includes at least one selected from sugars, cellulose and its derivatives, calcium phosphates, alkaline earth metal stearates, vegetable oils, nonionic surfactants, cationic surfactants, anionic surfactants, fatty alcohols, and cereal hydrolyzed solids.

[0021] According to an embodiment of the present invention, the dosage form of the composition includes at least one selected from oral liquid, powder, granule, capsule, tablet, and pill.

[0022] A seventh aspect of the present invention provides the use of the microorganism described in the first aspect, the fermentation broth described in the second aspect, the bacterial suspension described in the third aspect, or the composition described in the sixth aspect for preventing, alleviating, and / or treating diseases caused by pathogenic bacteria. As previously mentioned, the Akkermansia muciniphila VB202 provided by the present invention has good antibacterial ability and intestinal tolerance. Therefore, the microorganism and the fermentation broth, bacterial suspension, or composition containing the microorganism also have good antibacterial and intestinal tolerance, and can effectively prevent, alleviate, and / or treat diseases caused by pathogenic bacteria.

[0023] According to an embodiment of the present invention, the pathogenic bacteria is selected from at least one of Helicobacter pylori, Listeria monocytogenes, and Shigella.

[0024] The eighth aspect of the present invention provides a method for preventing, alleviating and / or treating related diseases caused by pathogenic bacteria infection in an individual. According to an embodiment of the present invention, the method comprises administering to the individual the microorganism described in the first aspect, the fermentation broth described in the second aspect, the bacterial suspension described in the third aspect, or the composition described in the sixth aspect. As mentioned above, the Akkermansia muciniphila VB202 provided by the present invention has good antibacterial ability and intestinal tolerance. Therefore, the microorganism and the fermentation broth, bacterial suspension or composition containing the microorganism also have good antibacterial and intestinal tolerance, and can help the body restore the balance of the microbiome. The method according to the embodiment of the present invention can effectively prevent, alleviate and / or treat related diseases caused by pathogenic bacteria infection.

[0025] According to an embodiment of the present invention, the pathogenic bacteria is selected from at least one of Helicobacter pylori, Listeria monocytogenes, and Shigella.

[0026] A ninth aspect of the present invention provides the use of the microorganism described in the first aspect, the fermentation broth described in the second aspect, the bacterial suspension described in the third aspect, or the composition described in the sixth aspect for preventing, alleviating, and / or treating hyperglycemia. As previously mentioned, the Akkermansia muciniphila VB202 provided by the present invention has a significant blood sugar-lowering effect. Therefore, the microorganism and the fermentation broth, bacterial suspension, or composition containing the microorganism also have significant blood sugar-lowering and intestinal tolerance capabilities, and can effectively prevent, alleviate, and / or treat hyperglycemia and related diseases caused by it, such as metabolic syndrome.

[0027] The tenth aspect of the present invention provides a method for preventing, alleviating and / or treating hyperglycemia in an individual. According to an embodiment of the present invention, the method comprises administering to the individual the microorganism described in the first aspect, the fermentation broth described in the second aspect, the bacterial suspension described in the third aspect, or the composition described in the sixth aspect. As previously mentioned, the Akkermansia muciniphila VB202 provided by the present invention has a good effect of lowering blood sugar. Therefore, the microorganism and the fermentation broth, bacterial suspension or composition containing the microorganism also have good blood sugar lowering and intestinal tolerance capabilities. The method according to the embodiment of the present invention can effectively prevent, alleviate and / or treat hyperglycemia and related diseases caused by it, such as metabolic syndrome, and at the same time has a non-gas-producing and synergistic anti-inflammatory effect.

[0028] The beneficial effects of the present invention compared to the prior art are as follows:

[0029] The Akkermansia muciniphila VB202 strain provided by the present invention exhibits strong intestinal tolerance and antibacterial properties. It can help restore the body's microbiome balance by inhibiting the growth of harmful bacteria without causing drug resistance, making it a safer alternative to antibiotics. Furthermore, this strain has hypoglycemic effects, is non-gas-producing, and exhibits synergistic anti-inflammatory effects. It can be used to prepare blood sugar control products, among other applications, and has broad application prospects.

[0030] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention.

[0031] Collection information:

[0032] Strain name: Akkermansia muciniphila VB202

[0033] Deposit date: August 29, 2023

[0034] Depository: General Microbiology Center of China Culture Collection Administration

[0035] Deposit number: CGMCC No.28295. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0037] FIG1 shows a bacterial colony photograph of the strain VB202 provided by the present invention on a culture medium;

[0038] FIG2 shows a Gram staining photograph of strain VB202 provided by the present invention;

[0039] FIG3 shows a microscopic photograph of the strain VB202 provided by the present invention;

[0040] FIG4 shows a phylogenetic tree of strain VB202 provided by the present invention;

[0041] FIG5 shows the acid resistance test results of the strain VB202 provided by the present invention;

[0042] FIG6 shows the bile salt tolerance test results of the strain VB202 provided by the present invention;

[0043] FIG7 shows the results of an intestinal fluid resistance test of the strain VB202 provided by the present invention;

[0044] FIG8 shows the blood sugar lowering test results of the strain VB202 provided by the present invention. DETAILED DESCRIPTION

[0045] Below, the scheme of the present invention will be explained in conjunction with embodiment.It will be understood by those skilled in the art that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.In the embodiment, if specific technology or conditions are not indicated, the technology or conditions described in the literature in this area or the product instructions are used.The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained by commercial purchase.

[0046] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, features defined as "first" or "second" may explicitly or implicitly include one or more of such features. Furthermore, in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0047] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0048] In order to make the present invention more easily understood, certain technical and scientific terms are specifically defined below. Unless otherwise clearly defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by those skilled in the art to which the present invention belongs.

[0049] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention, but not excluding other contents.

[0050] As used herein, the terms "optionally," "optional," or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0051] As used herein, the terms "treat" and "alleviate" refer to methods used to obtain a desired pharmacological and / or physiological effect. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, particularly humans, and includes: (a) preventing the occurrence of a disease or condition in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as arresting the progression of the disease; or (c) alleviating the disease, such as alleviating the symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or compound to an individual to treat, cure, alleviate, improve, reduce or inhibit the individual's disease, including but not limited to administering a drug containing a compound described herein to an individual in need.

[0052] As used herein, the term "carrier" includes any solvent, pharmaceutical stabilizer, or combination thereof, which are known to those skilled in the art. Except where any conventional carrier is incompatible with the active ingredient, its use in therapeutic or pharmaceutical compositions is encompassed.

[0053] According to an embodiment of the present invention, a first aspect of the present invention provides a microorganism, which is Akkermansia muciniphila VB202, which was deposited in the General Microbiology Center of the China Culture Collection Administration Committee on August 29, 2023, with a deposit number of CGMCC No. 28295. The Akkermansia muciniphila VB202 provided by the present invention has a strong antibacterial ability and can be used to replace antibiotics. At the same time, the strain has good blood sugar lowering ability and intestinal tolerance. The hypoglycemic drugs currently on the market have certain side effects, such as gas production, causing intestinal inflammation, etc., and the Akkermansia muciniphila VB202 provided by the present invention not only has the effect of lowering blood sugar, but also has no gas production and synergistic anti-inflammatory effects, and can be used to prepare blood sugar control products.

[0054] As used herein, "Akkermansia muciniphila VB202" and "Akkermansia muciniphila VB202", "VB202", and "strain VB202" are synonymous.

[0055] The Akkermansia muciniphila VB202 provided by the present invention has strong inhibitory ability against three pathogens, which are Helicobacter pylori, Listeria monocytogenes and Shigella.

[0056] The Akkermansia muciniphila VB202 provided by the present invention also exhibits excellent blood sugar-lowering ability and strong intestinal tolerance. Specifically, the strain exhibited minimal impact on bacterial survival after four hours in a 0.03% to 0.3% taurine solution, demonstrating its strong tolerance to taurine. It also survived for at least four hours in simulated gastric fluid at a pH of 3.0-6.0 and for four hours in simulated intestinal fluid at a pH of 6.8, with a 4-hour survival rate of 73.56%. This demonstrates that Akkermansia muciniphila VB202 exhibits excellent activity tolerance to the gastrointestinal environment and can function in the intestines for an extended period.

[0057] According to a specific embodiment of the present invention, the present invention provides a fermentation broth, which is fermented by the aforementioned microorganism.

[0058] According to a specific embodiment of the present invention, the present invention provides a bacterial suspension comprising the aforementioned microorganisms.

[0059] According to a specific embodiment of the present invention, the present invention provides the use of the aforementioned microorganisms, fermentation broth or bacterial suspension in the preparation of drugs, feeds, and additives that inhibit the activity of pathogens, or in the preparation of drugs, feeds, and additives that control or lower blood sugar.

[0060] According to a specific embodiment of the present invention, the present invention provides a composition comprising at least one of the aforementioned microorganisms, fermentation broth or bacterial suspension.

[0061] According to a specific embodiment of the present invention, the present invention provides the use of the aforementioned microorganisms, fermentation broth, bacterial suspension or composition in preventing, alleviating and / or treating diseases related to pathogen infection.

[0062] According to a specific embodiment of the present invention, the present invention provides a method for preventing, alleviating and / or treating related diseases caused by pathogenic bacteria infection in an individual, the method comprising administering the aforementioned microorganism, fermentation broth, bacterial suspension or the aforementioned composition to the individual.

[0063] According to a specific embodiment of the present invention, the present invention provides use of the aforementioned microorganism, fermentation broth, bacterial suspension or compound in preventing, alleviating and / or treating hyperglycemia.

[0064] According to a specific embodiment of the present invention, the present invention provides a method for preventing, alleviating and / or treating hyperglycemia in an individual, the method comprising administering the aforementioned microorganism, fermentation broth, bacterial suspension or composition to the individual.

[0065] Below, the scheme of the present invention will be explained in conjunction with embodiment.It will be understood by those skilled in the art that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.In the embodiment, if specific technology or conditions are not indicated, the technology or conditions described in the literature in this area or the product instructions are used.The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained by commercial purchase.

[0066] Example 1 Collection and identification of strains

[0067] 1. Isolation and purification of strains

[0068] The Akkermansia muciniphila VB202 provided by the present invention is isolated from the feces of healthy adults. The specific method is as follows: 0.5 g of feces is added to 10 mL of PBS solution containing 0.05% cysteine ​​to obtain a stock solution, and the stock solution is concentrated by 10 to 10 5 After doubling, the strains were inoculated into AKK enrichment medium and cultured in an anaerobic incubator at 37°C for 4 days with an inoculum size of 10%. DNA of the strains at different dilutions was extracted and gene amplification was performed using the 16S full-length primers 27F (SEQ ID NO: 2: 5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (SEQ ID NO: 3: 5'-TACGGCTACCTTGTTACGACTT-3'). PCR amplification products were detected on a 1.5% agarose gel. The tube with a positive PCR result and the highest dilution was selected and stored in BHI + 10% glycerol protective medium at -80°C.

[0069] The enrichment culture medium comprises (in W / V): 3.85% BHI culture medium, 0.25% mucin, 0.05% L-cysteine, and is sterilized at 115° C. for 20 minutes.

[0070] 2. Identification of strains

[0071] (1) Morphological characteristics:

[0072] The strain VB202 was cultured anaerobically at 37°C on an isolation medium for 4-5 days to form circular colonies with a diameter of 1-3 mm, a glossy milky white surface, and a light milky yellow center. The colony morphology is shown in FIG1 .

[0073] The separation medium comprises (in W / V): 3.85% BHI medium, 0.25% mucin, 0.05% L-cysteine, and 1.8% agar, and is sterilized at 115° C. for 20 minutes.

[0074] (2) Physiological and biochemical characteristics:

[0075] Clean slides were Gram-stained and microscopically examined to observe the microscopic morphology of the strain, as shown in Figures 2 and 3. Its utilization of various carbon sources is shown in Table 1, and its utilization of various nitrogen sources is shown in Table 2. The results showed that the strain could grow on carbon sources such as glucose, fructose, and maltose, and on nitrogen sources such as tryptone, polypeptone, and hydrolyzed casein. Its growth was slow in soybean meal, soybean meal meal, and yeast extract FM860, with later effects similar to those in soybean peptone OX and yeast extract 601. Furthermore, the strain did not utilize monosodium glutamate for growth, and malt extract had an inhibitory effect on the bacteria.

[0076] Table 1: Carbon source utilization of strain VB202

[0077] Note: +: can utilize this carbon source.

[0078] Table 2: Nitrogen source utilization of strain VB202

[0079] Note: +: can utilize the nitrogen source, -: cannot utilize the nitrogen source.

[0080] (3) 16S rDNA gene analysis

[0081] The obtained positive tube was concentrated with PBS solution for 10 to 10 5 The strains were plated on AKK isolation medium and cultured in an anaerobic chamber at 37°C for 4-5 days. Round single colonies with a diameter of less than 1 mm were selected and inoculated into AKK screening medium for culture. Their 16S rDNA was sequenced and analyzed. The sequencing results were compared using BLAST, specifically using a k-mer frequency fingerprinting method. First, k-mers of length 16 were extracted from each sequence, and their occurrences were counted to generate a k-mer frequency fingerprint. Next, the cosine distance was used to calculate the similarity between the k-mer frequency fingerprints of the two sequences. Using these distance values, a phylogenetic tree was constructed, as shown in Figure 4. It was found that the obtained strain was closely related to Akkermansia_muciniphila ATCC_BAA-835, and then formed a larger related group with Akkermansia_massiliensis Marseille-P6666 and PJKB_s GP22, showing that the obtained strain had a relatively close relationship with these species. Therefore, the strain was identified as Akkermansia muciniphila strain, numbered VB202, and was deposited in the General Microbiology Center of China Culture Collection Administration on August 29, 2023, with the deposit number CGMCC No.28295.

[0082] The primary screening culture medium comprises (in W / V): 3.85% BHI culture medium and 0.05% L-cysteine, and is sterilized at 115° C. for 20 minutes.

[0083] The 16S rDNA sequence of the strain is:

[0084] Example 2 Preparation of fermentation broth and physiological and biochemical characteristics of strain VB202

[0085] After gradient dilution of the glycerol tube containing strain VB202, spread it on AKK separation medium and culture it at 37℃ for 4-5 days. Select single colonies with a diameter of 1-3 mm and inoculate them into AKK enrichment medium. The inoculation volume is 10 7 CFU / mL, cultured at 37 ° C, 60-80 rpm for 24-30 h, when the pH of the seed solution was 5.5-6.8, the OD value was 0.8-1.5, and the AKK bacterial count was 5×10 8 ~2×10 9 CFU / ml, 3% of the inoculum was inoculated into a fermenter and cultured at 37°C and 60 rpm for 30-48 h. The physiological and biochemical characteristics were analyzed. The test results are shown in Tables 3-4.

[0086] Table 3: Physiological and biochemical characteristics of strain VB202

[0087] Note: + represents positive, - represents negative.

[0088] Table 4: Physiological and biochemical characteristics of strain VB202

[0089] Note: + represents positive, - represents negative.

[0090] Example 3 Evaluation of the gastrointestinal tolerance of strain VB202

[0091] Preparation of artificial simulated gastric fluid: 1% sodium chloride solution with pH values ​​of 1, 2, 3, 4, 5, and 6 was prepared, sterilized at 121°C for 30 min, and pepsin was added at 0.3%;

[0092] Artificial intestinal fluid: R22156-500ml, pH 6.8, Shanghai Yuanye Biotechnology;

[0093] Artificial simulated pancreatic juice: first prepare a 1% sodium chloride solution, add 0.03%, 0.1%, and 0.3% taurine choline, sterilize at 121°C for 30 minutes, and add 0.1% trypsin.

[0094] Preparation of bacterial suspension: Take the counted bacterial suspension (the viable bacteria count should reach about 10 9 ~10 10 After thawing, the cells were added to the aliquoted artificial simulated gastric juice, intestinal juice, and pancreatic juice at a volume of 10 μL bacterial suspension + 990 μL simulated solution. Mix well and culture anaerobically at 37°C. Samples were taken at 0 h, 2.5 h, and 4 h for viable bacterial counts, and the survival rates at 2.5 h and 4 h were calculated.

[0095] The results of the acid resistance test are shown in Figure 5. Strain VB202 died instantly in simulated gastric fluid at pH 1; all died within 2.5 hours in simulated gastric fluid at pH 2. However, it remained stable for 4 hours in simulated gastric fluid at pH 3-6. The results of the bile salt resistance test are shown in Figure 6. Strain VB202 remained stable for 4 hours in a 0.03% to 0.3% taurine system. The results of the intestinal fluid resistance test are shown in Figure 7. The survival rate of strain VB202 in simulated intestinal fluid was approximately 84.28% after 2.5 hours, and approximately 73.56% after 4 hours. These results indicate that strain VB202 exhibits excellent tolerance to the gastrointestinal environment.

[0096] Example 4 Antibacterial test of strain VB202

[0097] The antibacterial ability of the strain was tested using the double-layer plate culture method, and the growth of pathogenic bacteria in the upper layer was used to evaluate whether the strain VB202 had antibacterial ability. The specific method is as follows:

[0098] After thawing the frozen tube of activated strain VB202, inoculate it into AKK isolation medium and culture it anaerobically at 37℃ for 4-5 days. The inoculation volume is 2μl / point. The sterilized upper culture medium is cooled to 40℃-50℃, and different pathogens (Helicobacter pylori ATCC26695, Listeria monocytogenes ATCC19114, Shigella CMCC51252) are added (the final concentration of pathogens in the culture medium is about 10 6 CFU / ml) and evenly mixed. The culture medium containing pathogenic bacteria was added to the above-mentioned single bacterial plate at 7 ml / dish. After solidification, the culture was cultured according to the growth conditions of different pathogens. The presence and size of the inhibition zone around the VB202 single colony were observed to determine whether it had antibacterial ability. The results are shown in Table 5.

[0099] As shown in Table 5, Akkermansia muciniphila VB202 has the effect of inhibiting Helicobacter pylori, Listeria monocytogenes and Shigella.

[0100] Helicobacter pylori culture conditions include fetal bovine serum Brucella broth, microaerophilic culture at 37°C for 3 days. Listeria monocytogenes culture conditions include LB medium, aerobic culture at 37°C for 1 day. Shigella culture conditions include LB medium, aerobic culture at 37°C for 1 day.

[0101] Table 5: Evaluation of the antibacterial ability of strain VB202 against three pathogenic bacteria

[0102] Example 5 Hypoglycemic Test of Strain VB202

[0103] Twelve-week-old genetically deficient type 2 diabetes model dbdb male mice were selected. After one week of adaptation, blood glucose levels were measured by tail clipping and fasting blood glucose was measured once a week. After one month of feeding, 30 dbdb mice with fasting blood glucose levels greater than 11.1 mmol / L were selected and divided into two groups, each with 10 mice. One group was the control group, and the other group was given 10 doses of fresh live bacteria VB202 each time. 10 CFU were administered once every 12 hours for 3 weeks, and fasting blood glucose was measured once a week. As shown in Figure 8, strain VB202 had a good effect in lowering fasting blood glucose in the mouse model.

[0104] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", "some implementation plans" or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0105] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A microorganism, characterized in that The microorganism is Akkermansia muciniphila VB202, which was deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on August 29, 2023, with the deposit number CGMCC No.28295.

2. The microorganism according to claim 1, characterized in that The microorganism has a 16S rDNA sequence as shown in SEQ ID NO:

1.

3. A fermentation broth, characterized in that: The fermentation broth is fermented by the microorganism according to claim 1 or 2.

4. A bacterial suspension, characterized in that The bacterial suspension comprises the microorganism according to claim 1 or 2.

5. Use of the microorganism according to claim 1 or 2, the fermentation liquid according to claim 3 or the bacterial suspension according to claim 4 in the preparation of medicines, feeds and additives for inhibiting the activity of pathogenic bacteria.

6. The use according to claim 5, characterized in that The pathogenic bacteria is selected from at least one of Helicobacter pylori, Listeria monocytogenes and Shigella.

7. Use of the microorganism according to claim 1 or 2, the fermentation liquid according to claim 3 or the bacterial suspension according to claim 4 in the preparation of medicines, feeds and additives for controlling or lowering blood sugar.

8. A composition, characterized in that The composition comprises at least one of the microorganism according to claim 1 or 2, the fermentation broth according to claim 3, and the bacterial suspension according to claim 4.

9. The composition according to claim 8, characterized in that The composition further comprises an excipient and / or a carrier.

10. The composition according to claim 9, characterized in that The excipient includes at least one selected from a binder, a disintegrant, a lubricant, a glidant, a stabilizer, a filler, a diluent, and a sustained-release agent; Optionally, the carrier comprises at least one selected from sugars, cellulose and its derivatives, calcium phosphates, stearic acid alkaline earth metal salts, vegetable oils, nonionic surfactants, cationic surfactants, anionic surfactants, fatty alcohols, and hydrolyzed cereal solids; Optionally, the dosage form of the composition includes at least one selected from oral liquid, powder, granule, capsule, tablet, and pill.

11. Use of the microorganism according to claim 1 or 2, the fermentation broth according to claim 3, the bacterial suspension according to claim 4 or the composition according to any one of claims 8 to 10 in preventing, alleviating and / or treating diseases related to pathogenic bacteria infection.

12. A method for preventing, alleviating and / or treating diseases caused by pathogenic bacteria infection in an individual, characterized in that: The method comprises administering to the individual the microorganism according to claim 1 or 2, the fermentation broth according to claim 3, the bacterial suspension according to claim 4, or the composition according to any one of claims 8 to 10.

13. Use of the microorganism according to claim 1 or 2, the fermentation broth according to claim 3, the bacterial suspension according to claim 4 or the composition according to any one of claims 8 to 10 in preventing, alleviating and / or treating hyperglycemia.

14. A method for preventing, alleviating and / or treating hyperglycemia in an individual, characterized in that: The method comprises administering to the individual the microorganism according to claim 1 or 2, the fermentation broth according to claim 3, the bacterial suspension according to claim 4, or the composition according to any one of claims 8 to 10.

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