WO2025118641A1 - Akkermansia muciniphila vb202 and use thereof - Google Patents

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

Priority information

This application claims the priority of Chinese patent application No. 202311658889.8, filed with the Chinese Patent Office on December 5, 2023, and entitled “A Mucinophilic Akkermansia VB202 and Its Applications”, the entire contents of which are incorporated by reference into this application.

Technical Field

The invention relates to the technical field of microorganisms, and in particular to Akkermansia muciniphila VB202 and applications thereof.

Background Art

Akkermansia muciniphila (AKK) is a new intestinal microorganism first discovered in the feces of healthy people by Derrien et al. in 2004. It belongs to the Akkermansia genus in the Verrucomicrobiaceae family of the phylum Verrucomicrobia. It uses mucin as its only nitrogen and carbon source and colonizes in the intestinal mucosa. Akkermansia muciniphila is widely distributed in the intestines of healthy infants and adults. It has been stably colonized in the intestines one year after birth and accounts for 1% to 3% of the total number of intestinal microorganisms in adults. It is an important component of the intestinal flora. Studies have found that the content of this bacteria in the intestines of patients with various diseases is significantly reduced, and the content of this bacteria is correlated with the disease condition. Therefore, it is expected to be developed into the next generation of probiotics after bifidobacteria and lactobacilli (O'Toole PW, Marchesi JR, Hill C. Next-generation probiotics: the spec-trum from probiotics to live biotherapeutics [J]. Nat Microbiol, 2017, 2: 17057).

Different types of pathogens can not only cause food safety problems, but also infect humans and animals and cause related diseases and even threaten life. At present, antibiotics or chemical synthetic drugs are mainly used to inhibit or kill pathogens to achieve effective prevention and control. However, due to the increasing drug resistance of pathogens, irreversible damage such as chemical drug residues in the body and environmental pollution has occurred frequently, which has posed a severe challenge to human health and the sustainable development of social economy. Announcement No. 194 issued by my country's Ministry of Agriculture and Rural Affairs in July 2019 pointed out that from January 1, 2020, the addition of antibiotics to feed in my country will be completely banned, and the research on antibiotic substitutes has become increasingly urgent. Probiotics can produce related metabolites that have an inhibitory effect on pathogens during growth and reproduction, and can improve the acidic environment of the animal's intestines, optimize the dynamic balance of intestinal microorganisms, stimulate the body's intestinal mucosal immunity, inhibit the colonization of harmful bacteria, and improve the body's immunity, thereby improving animal production performance and health level. There is a broad development prospect in the natural prevention and control of pathogens.

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.

Summary of the invention

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

To this end, the first aspect of the present invention provides a microorganism, which 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.

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, and help the body 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, the strain has a blood sugar lowering effect and can be used to prepare blood sugar control products.

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

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

The third aspect of the present invention provides a bacterial suspension, which includes the microorganisms described in the first aspect.

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 drugs, feeds, and additives for inhibiting the activity of pathogenic bacteria.

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

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.

The sixth aspect of the present invention provides a composition, which comprises at least one of the microorganism described in the first aspect, the fermentation broth described in the second aspect, and the bacterial suspension described in the third aspect.

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

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.

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 hydrolyzed cereal solids.

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.

The 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 in preventing, alleviating and/or treating related diseases caused by pathogenic bacteria infection. 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 effectively prevent, alleviate and/or treat related diseases caused by pathogenic bacteria infection.

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

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 muciniphilic Akkermansia 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, which can help the body restore the balance of the microbiome. The method according to an embodiment of the present invention can effectively prevent, alleviate and/or treat related diseases caused by pathogenic bacteria infection.

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

The 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 in preventing, alleviating and/or treating hyperglycemia. As mentioned above, 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, and can effectively prevent, alleviate and/or treat hyperglycemia and related diseases caused by it, such as metabolic syndrome.

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 mentioned above, the muciniphilic Akkermansia 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. 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 no gas production and synergistic anti-inflammatory effects.

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

The Akkermansia muciniphila VB202 provided by the present invention has strong intestinal tolerance and antibacterial ability, can help the body restore the balance of the microbiome by inhibiting the growth of harmful bacteria, and will not cause the generation of drug resistance, and can be used as a safer alternative to antibiotics. In addition, the strain also has a hypoglycemic effect, does not produce gas, and has a synergistic anti-inflammatory effect, can be used to prepare blood sugar control products, etc., and has a broad application prospect.

Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention.

Collection information:

Strain name: Akkermansia muciniphila VB202

Deposit date: August 29, 2023

Depository: China National Microbiological Culture Collection Administration General Microbiology Center

Collection number: CGMCC No.28295.

BRIEF DESCRIPTION OF THE DRAWINGS

The above and/or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

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

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

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

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

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

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

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

FIG8 shows the blood sugar lowering test results of the strain VB202 provided by the present invention.

DETAILED DESCRIPTION

The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

The endpoints and any values of the ranges disclosed in this article 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 endpoint values of each range, the endpoint values of each range and the 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 as specifically disclosed in this article.

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 meanings commonly understood by those skilled in the art to which the present invention belongs.

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.

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.

As used herein, the terms "treatment" and "alleviation" both refer to the use of drugs to obtain the desired pharmacological and/or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and/or may be 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 an individual who is susceptible to the disease but has 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 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 an individual's disease, including but not limited to administering a drug containing a compound described herein to an individual in need.

As used herein, the term "carrier" includes any solvent, drug 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 treatment or pharmaceutical compositions is contemplated.

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 China Microorganism Culture Collection Administration 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, 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.

In this article, “Akkermansia muciniphila VB202” and “Akkermansia muciniphila VB202”, “VB202”, and “strain VB202” are synonymous.

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

The muciniphilic Akkermansia VB202 provided by the present invention also has good blood sugar lowering ability and strong intestinal tolerance. Specifically, the muciniphilic Akkermansia VB202 provided by the present invention has little effect on bacterial survival in a 0.03% to 0.3% oxcholine solution for 4 hours, indicating that the strain VB202 has a strong tolerance to oxcholine. It can survive for at least 4 hours in simulated gastric fluid at pH 3.0-6.0, and can survive for 4 hours in simulated intestinal fluid at pH 6.8, with a 4-hour survival rate of 73.56%, indicating that the muciniphilic Akkermansia VB202 has a good active tolerance to the gastrointestinal environment and can play a role in the intestine for a long time.

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

According to a specific embodiment of the present invention, the present invention provides a bacterial suspension, which includes the aforementioned microorganisms.

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, additives for inhibiting the activity of pathogenic bacteria, or in the preparation of drugs, feeds, additives for controlling or lowering blood sugar.

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.

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 pathogenic bacteria infection.

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.

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.

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.

The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

Example 1 Collection and identification of strains

1. Isolation and purification of strains

The Akkermansia muciniphila VB202 provided by the present invention is isolated from the feces of healthy adults, and 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 times and then inoculated into AKK enrichment medium respectively, and placed in an anaerobic box at 37°C for enrichment culture for 4 days, with an inoculation amount of 10%. DNA of strains with different dilutions is extracted, and gene amplification is performed using 16S full-length primers 27F (SEQ ID NO: 2: 5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (SEQ ID NO: 3: 5'-TACGGCTACCTTGTTACGACTT-3'), and the PCR amplification product is detected by 1.5% agarose gel, and a tube with a positive PCR identification result and the highest dilution is selected, and the positive tube is stored in BHI+10% glycerol Protective agent, stored at -80 ℃.

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

2. Identification of strains

(1) Morphological characteristics:

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

The separation medium is composed of (by 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.

(2) Physiological and biochemical characteristics:

Take a clean slide for Gram staining and microscopic examination to observe the microscopic morphology of the strain, as shown in Figures 2 and 3. Its utilization of different carbon sources is shown in Table 1, and its utilization of different nitrogen sources is shown in Table 2. The results show that the strain can grow using carbon sources such as glucose, fructose, and maltose, and can also grow using nitrogen sources such as tryptone, polypeptone, and hydrolyzed casein. It takes effect slowly in soybean powder, soybean meal powder, and yeast extract powder FM860, and the later effect is similar to that of soybean peptone OX and yeast powder 601. At the same time, the strain does not use monosodium glutamate for growth, and malt extract powder has an inhibitory effect on the bacteria.

Table 1: Carbon source utilization of strain VB202

Note: +: can utilize the carbon source.

Table 2: Nitrogen source utilization of strain VB202

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

(3) 16S rDNA gene analysis

The obtained positive tube was concentrated 10 to 10 ⁵ times with PBS solution, spread on AKK separation medium, and cultured in an anaerobic box at 37°C for 4-5 days. A circular single colony with a diameter of 1 mm was selected and inoculated in AKK primary screening medium for culture, and 16S rDNA sequencing analysis was performed on it. The sequencing results were compared by BLAST, specifically using a method based on k-mer frequency fingerprints. First, a k-mer with a length of 16 was extracted from each sequence, and its number of occurrences was counted to generate a k-mer frequency fingerprint. Then, 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 Microorganism Culture Collection Administration on August 29, 2023, with the deposit number CGMCC No.28295.

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

The 16S rDNA sequence of the strain is:

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

The glycerol tube containing strain VB202 was diluted gradiently and then spread on AKK separation medium. It was cultured at 37°C for 4-5 days. A single colony with a diameter of 1-3 mm was selected and inoculated into AKK enrichment medium with an inoculum of 10 ⁷ CFU/mL. It was cultured at 37°C and 60-80 rpm for 24-30 h. When the seed solution pH 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 it was 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.

Table 3: Physiological and biochemical characteristics of strain VB202

Note: + is positive, - is negative.

Table 4: Physiological and biochemical characteristics of strain VB202

Note: + is positive, - is negative.

Example 3 Evaluation of the gastrointestinal tolerance of strain VB202

Preparation of artificial simulated gastric juice: prepare 1% sodium chloride solution with pH values of 1, 2, 3, 4, 5, and 6, sterilize at 121°C for 30 min, and add 0.3% pepsin;

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

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

Preparation of bacterial suspension: Take the counted bacterial suspension (the live bacteria count needs to reach about 10 ⁹ to 10 ¹⁰ CFU/ml), thaw it, and add it to the above-mentioned artificial simulated gastric juice, intestinal juice and pancreatic juice. The amount added is: 10 μL bacterial suspension + 990 μL simulated liquid. After mixing, culture anaerobically at 37°C, take samples at 0h, 2.5h, and 4h for live bacteria count, and calculate the survival rate at 2.5h and 4h.

The results of the acid resistance test are shown in Figure 5. Among them, strain VB202 died immediately in simulated gastric fluid with a pH of 1; all died in simulated gastric fluid with a pH of 2 for 2.5 hours, and could survive stably in simulated gastric fluid with a pH of 3 to 6 for 4 hours. The results of the bile salt resistance test are shown in Figure 6. Among them, strain VB202 could survive stably in a 0.03% to 0.3% oxcholine system for 4 hours. The results of the intestinal fluid resistance test are shown in Figure 7. Among them, the survival rate of strain VB202 in simulated intestinal fluid for 2.5 hours was about 84.28%, and the survival rate for 4 hours was about 73.56%. In summary, strain VB202 has a good active tolerance to the gastrointestinal environment.

Example 4 Antibacterial test of strain VB202

The antibacterial ability of the strain was tested by double-layer plate culture method, and the antibacterial ability of strain VB202 was evaluated based on the growth of pathogenic bacteria in the upper layer. The specific method is as follows:

After the activated strain VB202 cryotube was thawed, it was inoculated in AKK separation medium and cultured anaerobically at 37℃ for 4-5 days, with the inoculation amount of 2μl/point. The sterilized upper culture medium was cooled to 40℃-50℃, and different pathogenic bacteria (Helicobacter pylori ATCC26695, Listeria monocytogenes ATCC19114, Shigella CMCC51252) were added (the final concentration of pathogenic bacteria in the culture medium was about 10 ⁶ CFU/ml) and mixed evenly. The culture medium containing pathogenic bacteria was added to the above-mentioned single bacterial plate at 7ml/dish. After solidification, it was cultured according to the growth conditions of different pathogenic bacteria, and the inhibition zone and the 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.

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

The culture conditions for Helicobacter pylori are: fetal bovine serum Brucella broth medium, microaerobic culture at 37°C for 3 days. The culture conditions for Listeria monocytogenes are: LB medium, aerobic culture at 37°C for 1 day. The culture conditions for Shigella are: LB medium, aerobic culture at 37°C for 1 day.

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

Example 5 Blood sugar lowering test of strain VB202

12-week-old genetically defective type 2 diabetes model animal dbdb male mice were selected. After one week of adaptation, the tail was cut and blood was collected to measure blood sugar levels. Fasting blood sugar was measured once a week. After one month of feeding, 30 dbdb mice with fasting blood sugar levels greater than 11.1mmol/L were selected and divided into 2 groups, each with 10 mice, one group was a control, and the other group was administered with fresh live bacteria VB202 at a dosage of 10 ¹⁰ CFU every 12 hours for 3 weeks, and fasting blood sugar was measured once a week. The results are shown in Figure 8. The strain VB202 has a good effect on reducing fasting blood sugar in the mouse model.

In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", "some implementation schemes" or "some examples" etc. means 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 representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims (14)

A microorganism, characterized in that the microorganism is Akkermansia muciniphila VB202, which was deposited in the General Microbiology Center of China Culture Collection Administration on August 29, 2023, with the deposit number CGMCC No. 28295. The microorganism according to claim 1 is characterized in that the microorganism has a 16S rDNA sequence as shown in SEQ ID NO:1. A fermentation broth, characterized in that the fermentation broth is fermented by the microorganism according to claim 1 or 2. A bacterial suspension, characterized in that the bacterial suspension comprises the microorganism according to claim 1 or 2. 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. The use according to claim 5 is characterized in that the pathogenic bacteria is selected from at least one of Helicobacter pylori, Listeria monocytogenes, and Shigella. 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 drugs, feeds and additives for controlling or lowering blood sugar. A composition, characterized in that the composition comprises at least one of the microorganisms according to claim 1 or 2, the fermentation broth according to claim 3, and the bacterial suspension according to claim 4. The composition according to claim 8, characterized in that the composition further comprises an excipient and/or a carrier. The composition according to claim 9, characterized in that the excipient comprises 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. 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. A method for preventing, alleviating and/or treating related diseases caused by pathogenic bacteria infection in an individual, characterized in that it 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. The microorganism according to claim 1 or 2, the fermentation liquid according to claim 3, the bacteria according to claim 4 Use of the suspension or the composition according to any one of claims 8 to 10 in preventing, alleviating and/or treating hyperglycemia. A method for preventing, alleviating and/or treating hyperglycemia in an individual, characterized in that it 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.