Bifidobacterium animalis subsp. lactis VB301 and use thereof

Abstract

Disclosed in the present invention is Bifidobacterium animalis subsp. lactis VB301 and a use thereof. The strain is preserved in China General Microbiological Culture Collection Center on 24 July 2023, and has the preservation number of CGMCC No. 27998. The Bifidobacterium animalis subsp. lactis VB301 of the present invention has a remarkable bile salt reduction capability, can be used for preparing cholesterol-lowering drugs, has relatively strong intestinal tolerance and antibacterial capability, and has large prospects of application in the preparation of drugs.

Description

Animal Bifidobacterium lactis subspecies VB301 and its application Technical Field

[0001] The present invention relates to the field of microorganisms, and in particular to animal Bifidobacterium lactis subspecies VB301 and applications thereof. Background Art

[0002] Cholesterol is a crucial lipid in the human body. At certain concentrations, it provides essential functions for the body. However, excessive cholesterol intake can lead to its accumulation in blood vessel walls, forming cholesterol plaques, which can cause a variety of cardiovascular diseases. Currently, cardiovascular and cerebrovascular diseases are the leading cause of death, and their incidence is increasing year by year. Therefore, lowering cholesterol levels has become an important approach to preventing cardiovascular disease.

[0003] In the liver, cholesterol's primary fate is conversion to bile acids. Glycocholic acid and taurocholic acid, derived from bile acids, are the primary bile acids in humans. Bile acids provide a crucial excretion pathway for cholesterol metabolism, with one-third of cholesterol catabolism occurring through bile acid synthesis. Bile acids can be divided into two categories: free bile acids such as cholic acid, deoxycholic acid, chenodeoxycholic acid, and lithocholic acid; and conjugated bile acids, formed by free bile acids bound to glycine or taurine. Six major types of conjugated bile acids exist in the human body: glycochenodeoxycholic acid (26%), glycocholic acid (26%), glycodeoxycholic acid (22%), taurocholic acid (9%), taurodeoxycholic acid (9%), and taurochenodeoxycholic acid (9%). Bile acids exist as sodium or potassium salts, known as bile salts.

[0004] The mechanisms proposed by probiotics at home and abroad for lowering cholesterol include: (1) probiotics adsorb cholesterol; (2) bound bile salts are hydrolyzed into free bile salts by the bile salt hydrolase of probiotics. Free bile salts are not easily absorbed by the intestine due to their low solubility. Cholesterol and free bile salts in the body co-precipitate in an acidic environment, thereby lowering serum cholesterol levels; (3) probiotics ferment carbohydrates from intestinal food to produce propionic acid, which inhibits the biosynthesis of cholesterol in the liver; (4) probiotics inhibit the formation of cholesterol emulsified micelles.

[0005] The prior art discloses a variety of animal Bifidobacterium lactis subspecies, but none of them has the ability to simultaneously degrade the six main bile salts bound to the human body. The six bile salts are sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), sodium glycocholate (GC), sodium glycochenodeoxycholate (GCDC), and sodium glycodeoxycholate (GDC). In addition, the various animal Bifidobacterium lactis subspecies disclosed in the prior art have no antibacterial effect or the antibacterial effect is not outstanding.

[0006] Summary of the Invention

[0007] The present invention aims to solve at least one of the technical problems existing in the prior art to at least a certain extent.

[0008] To this end, the first aspect of the present invention provides a Bifidobacterium animalis subsp. lactis VB301, which was deposited in the General Microbiology Center of the China Culture Collection Administration on July 24, 2023, with a deposit number of CGMCC No. 27998.

[0009] The animal Bifidobacterium lactis subspecies provided by the present invention has the ability to simultaneously degrade six types of bound bile salts, intestinal tolerance and antibacterial ability. On the one hand, it can be used to prevent and / or alleviate and / or treat related diseases caused by high cholesterol. On the other hand, it can prevent and / or alleviate and / or treat related diseases caused by at least one of Escherichia coli, Staphylococcus aureus, Salmonella, Listeria, Shigella, Enterococcus faecalis, and Clostridium perfringens, including infectious diseases such as urinary tract infection, purulent abdominal infection, sepsis, endocarditis, diarrhea and fever.

[0010] A second aspect of the present invention provides a fermentation supernatant, comprising the metabolites of the animal Bifidobacterium lactis subsp. lactis VB301 described in the first aspect.

[0011] A third aspect of the present invention provides a bacterial suspension, comprising the Bifidobacterium animalis subsp. lactis VB301 described in the first aspect.

[0012] The fourth aspect of the present invention provides use of the animal Bifidobacterium lactis subsp. VB301 described in the first aspect, the fermentation supernatant described in the second aspect, or the bacterial suspension described in the third aspect in the preparation of a medicament for preventing and / or treating diseases related to high cholesterol.

[0013] According to an embodiment of the present invention, the high cholesterol-induced related diseases include any one of hypercholesterolemia, cardiovascular disease, arteriosclerosis and peripheral vascular disease.

[0014] According to a specific embodiment of the present invention, the drug is capable of degrading six bile salts, and the six bile salts are sodium taurocholate, sodium taurochenodeoxycholate, sodium taurodeoxycholate, sodium glycocholate, sodium glycochenodeoxycholate, and sodium glycodeoxycholate.

[0015] The fifth aspect of the present invention provides the use of the animal Bifidobacterium lactis subspecies VB301 described in the first aspect or the bacterial suspension described in the third aspect in the preparation of a medicine for related diseases caused by at least one of Staphylococcus aureus, Escherichia coli, Salmonella Paratyphi B, Listeria monocytogenes, Shigella dysenteriae, Enterococcus faecalis, and Clostridium perfringens.

[0016] According to a specific embodiment of the present invention, the related diseases caused by Staphylococcus aureus, Escherichia coli, Salmonella Paratyphi B, Listeria monocytogenes, Shigella dysenteriae, Enterococcus faecalis, and Clostridium perfringens include infectious diseases such as urinary tract infection, purulent abdominal infection, sepsis, endocarditis, diarrhea and fever.

[0017] The sixth aspect of the present invention provides a medicine comprising at least one of the animal Bifidobacterium lactis subsp. VB301 described in the first aspect, the fermentation supernatant described in the second aspect, and the bacterial suspension described in the third aspect.

[0018] According to an embodiment of the present invention, the medicament further contains 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 drug includes at least one selected from oral liquid, powder, granule, capsule, tablet, and pill.

[0022] According to an embodiment of the present invention, when Bifidobacterium animalis subsp. lactis VB301 takes effect, it may exist in the form of living cells or in the form of non-living cells.

[0023] According to an embodiment of the present invention, the living cells refer to cells of Bifidobacterium animalis subspecies lactis VB301 that have the ability to metabolize, reproduce or replicate, and the non-living cells refer to cells that do not have the ability to metabolize, reproduce and replicate, including but not limited to dry bacteria (such as freeze-dried powder).

[0024] In addition, the present invention provides a microbial agent, which contains the aforementioned Bifidobacterium animalis subsp. lactis VB301, the aforementioned fermentation supernatant or the aforementioned bacterial suspension. The microbial agent of the present invention is used to prevent and / or treat related diseases caused by high cholesterol.

[0025] The present invention also provides use of the aforementioned Bifidobacterium animalis subsp. lactis VB301 and / or the aforementioned fermentation supernatant in preparing health products or foods, wherein the health products or foods are used to prevent and / or treat related diseases caused by high cholesterol.

[0026] 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.

[0027] Collection information:

[0028] Strain name: VB301

[0029] Deposit date: July 24, 2023

[0030] Depository: China General Microbiological Culture Collection Center (CGMCC)

[0031] Accession number: CGMCC No.27998 BRIEF DESCRIPTION OF THE DRAWINGS

[0032] 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:

[0033] Figure 1 shows the microscopic morphology of Bifidobacterium animalis subsp. lactis in Example 1 of the present invention, wherein A in Figure 1 represents the morphology of a single Bifidobacterium animalis subsp. lactis VB301 under a microscope, and B in Figure 1 represents the morphology of clustered Bifidobacterium animalis subsp. lactis VB301 under a microscope. DETAILED DESCRIPTION

[0034] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0035] 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.

[0036] In this document, the terms “contain”, “include” or “include” are open expressions, that is, they include the contents specified in the present invention but do not exclude other contents.

[0037] 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.

[0038] As used herein, "prevent" and "prevent" are used interchangeably. These terms refer to an approach for obtaining a beneficial or desired result, including but not limited to a prophylactic benefit. To obtain a "prophylactic benefit," Lactobacillus muciniphila ferments or a product containing the same can be administered to a subject at risk for developing a particular disease, or to a subject reporting one or more physiological symptoms of a disease, even though a diagnosis of the disease may not have yet been made.

[0039] 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.

[0040] In this article, "acceptable in health foods" refers to substances or compositions that can be consumed by humans, which may be adjusted according to the health food requirements of different countries.

[0041] As used herein, "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal to be treated therewith. Preferably, "pharmaceutically acceptable" as used herein means approved by federal regulatory agencies or national governments or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeia for use in animals, particularly humans.

[0042] As used herein, the term "pharmaceutically acceptable 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 treatment or pharmaceutical compositions is encompassed.

[0043] In this article, the term "pharmaceutically acceptable excipients" may include sugars, including monosaccharides or polysaccharides, such as lactose, sucrose, mannitol and sorbitol; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose and methyl cellulose; calcium phosphates, such as dicalcium phosphate and tricalcium phosphate; sodium sulfate; calcium sulfate; polyvinyl pyrrolidone; polyvinyl alcohol; stearic acid; alkaline earth metal stearates, such as magnesium stearate and calcium stearate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil; nonionic, cationic and anionic surfactants; ethylene glycol polymers; fatty alcohols; and cereal hydrolyzed solids and other non-toxic compatible fillers, binders, disintegrants, buffers, preservatives, antioxidants, lubricants, colorants, etc., which are commonly used excipients in pharmaceutical preparations.

[0044] According to a specific embodiment of the present invention, the present invention provides Bifidobacterium animalis subsp. lactis VB301. According to an embodiment of the present invention, the deposit number of Bifidobacterium animalis subsp. lactis is CGMCC No. 27998. The Bifidobacterium animalis subsp. lactis VB301 of the present invention has a significant bile salt-lowering ability and exhibits strong intestinal tolerance and antibacterial properties.

[0045] As used herein, "Bifidobacterium animalis subsp. lactis VB301" and "Bifidobacterium animalis subsp. lactis VB301", "VB301" and "strain VB301" are synonymous.

[0046] The animal Bifidobacterium lactis subspecies VB301 obtained by screening according to the present invention has a significant bile salt degrading ability compared with other animal Bifidobacterium lactis subspecies. The strain VB301 according to the present invention can degrade six bile salts, namely sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), sodium glycocholate (GC), sodium glycochenodeoxycholate (GCDC), and sodium glycodeoxycholate (GDC).

[0047] The present invention discloses a strain of Bifidobacterium animalis subsp. lactis VB301 that has a stronger intestinal tolerance than other Bifidobacterium animalis subsp. lactis. The strain VB301 is substantially unaffected by exposure to a 0.03% to 0.3% taurine solution for 4 hours, while maintaining a 91% survival rate after exposure to 0.3% taurine for 4 hours, indicating that the strain VB301 has a strong tolerance to taurine. Furthermore, the strain can survive in simulated intestinal fluid at a pH of 8.0 for 2-4 hours, with a 63% survival rate after 4 hours, indicating that the strain has a good active tolerance to the intestinal environment and can function in the intestinal tract for a long time.

[0048] The animal Bifidobacterium lactis subspecies VB301 of the present invention has stronger antibacterial ability than other animal Bifidobacterium lactis subspecies. Animal Bifidobacterium lactis subspecies VB301 has antibacterial effect on 7 pathogenic bacteria, and the 7 pathogenic bacteria are Escherichia coli, Staphylococcus aureus, Salmonella, Listeria, Shigella, Enterococcus faecalis, and Clostridium perfringens.

[0049] According to a specific embodiment of the present invention, a fermentation supernatant is provided, comprising: Bifidobacterium animalis subsp. lactis VB301 and / or metabolites of Bifidobacterium animalis subsp. lactis VB301. The Bifidobacterium animalis subsp. lactis VB301 of the present invention has a significant bile salt-lowering ability and exhibits strong intestinal tolerance and antibacterial properties.

[0050] It should be noted that “fermentation supernatant” refers to the supernatant after centrifugation of the fermentation broth, which may contain Bifidobacterium animalis subsp. lactis VB301 or its metabolites, or both. Bifidobacterium animalis subsp. lactis VB301 and its metabolites.

[0051] According to a specific embodiment of the present invention, a microbial agent is provided, comprising the aforementioned Bifidobacterium animalis subsp. lactis VB301 and / or the aforementioned fermentation supernatant. The microbial agent of the present invention is used to prevent and / or treat diseases related to high cholesterol.

[0052] It should be noted that the microbial agent of the present invention may be a liquid microbial agent, including but not limited to fermentation broth, etc., or a solid microbial agent, including but not limited to freeze-dried powder, etc.

[0053] According to an embodiment of the present invention, the Bifidobacterium animalis subsp. lactis VB301 exists in the form of living cells and / or non-living cells.

[0054] As used herein, "living cells" refer to cells that have the ability to metabolize, reproduce, or replicate.

[0055] For example, the living cells may be immobilized cells. Herein, "immobilized cells" refer to Bifidobacterium animalis subsp. lactis VB301 immobilized on a carrier, which can carry out life activities such as growth, development, reproduction, inheritance and metabolism within a certain spatial range.

[0056] Herein, "non-viable cells" refer to cells that do not have the ability to metabolize, reproduce, and replicate, including but not limited to dried bacteria. Exemplarily, the microbial agent is a freeze-dried powder.

[0057] As a specific embodiment, the Bifidobacterium animalis subspecies lactis VB301 exists in the form of living cells, dried bacteria, immobilized cells or any other forms.

[0058] As a specific embodiment, the dry bacteria are obtained by freeze-drying the animal Bifidobacterium lactis subspecies VB301.

[0059] According to an embodiment of the present invention, the present invention provides a medicine, health product, or food containing Bifidobacterium animalis subsp. lactis VB301. Preferably, the medicine, health product, or food of the present application is a live bacterial preparation for preparing cholesterol-lowering preparations. It should be noted that the medicine or health care product or food of the present application can be various preparations of a single active animal Bifidobacterium lactis subspecies VB301, or can be used in conjunction with other active ingredients, as long as the activity is not affected by each other; further, the best case of the medicine or health care product or food is that each active ingredient can have functional complementarity or a promoting effect, for example, animal Bifidobacterium lactis subspecies VB301 can be combined with other probiotics to make a composite probiotic tablet, which can be used to prevent and / or alleviate and / or treat the related diseases caused by high cholesterol on the one hand, and can prevent and / or alleviate and / or treat the related diseases caused by at least one of Escherichia coli, Staphylococcus aureus, Salmonella, Listeria, Shigella, Enterococcus faecalis, and Clostridium perfringens, including infectious diseases, specifically, can be determined according to the activity and components of the composite probiotic tablet, which is not limited here. Optionally, the medicine or health care product or food further includes a pharmaceutically acceptable carrier, a pharmaceutically acceptable excipient, an acceptable carrier or excipient in food, or an acceptable carrier or excipient in health food. It should be noted that when preparing live bacteria preparations, carriers or excipients are usually required, as long as the added carriers or excipients do not inhibit each other or cause adverse side effects with Bifidobacterium animalis subsp. lactis VB301. The medicine, health product or food can be in the form of powder, tablet, beverage or capsule.

[0060] 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.

[0061] Example 1: Strain collection and identification

[0062] Ten portions of fresh feces from healthy infants (3-5 years old), 10 portions of fresh feces from adults, and 3 portions of breast milk were collected. 0.5 g of feces and 1 mL of breast milk were added to 5 mL of Bifidobacterium BS liquid culture medium, respectively, and cultured under anaerobic conditions at 37°C for 12-16 h. The suspension was then homogenized by oscillation to prepare a bacterial suspension. 1 mL was taken for gradient dilution to prepare different gradients of diluted bacterial suspension, which were spread on MRS agar plates containing 50 μg / mL mupirocin and 5% serum (V / V). The suspension was anaerobically cultured at 37°C for 48-72 h. 130 colonies with different morphologies were picked, streaked, purified, and then DNA was extracted. Specific primers were used for PCR amplification and 16s rDNA sequencing was performed. The corresponding strains were stored in glycerol tubes. After DNA extraction, 16s rDNA full sequence sequencing was performed. The 16s sequencing results were compared by BLAST, and 18 strains of bifidobacteria were finally identified and isolated, including 1 strain of Bifidobacterium adolescentis, 11 strains of Bifidobacterium animalis subsp. lactis, 2 strains of Bifidobacterium bifidum, 2 strains of Bifidobacterium breve, and 2 strains of Bifidobacterium longum. The results are shown in Table 1.

[0063] Table 1: Strain collection and identification results

[0064] The 18 strains were tested for their ability to degrade six bile salts: sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), sodium glycocholate (GC), sodium glycochenodeoxycholate (GCDC), and sodium glycodeoxycholate (GDC).

[0065] Six different bile salts were added to BBL liquid culture medium at concentrations of 2-4 mmol / L. Activated glycerol tubes of each strain were removed from a -80°C freezer, thawed, and inoculated into BBL liquid culture medium at a 1-3% inoculum rate. The culture was then incubated at 37°C for 24 hours in an anaerobic workstation to obtain the activated bacterial solution. 1 mL of the culture medium was then centrifuged and the supernatant was collected for HPLC analysis of the bile salt content. The bile salt degradation capacity of each strain was assessed by calculating the bile salt degradation rate. A higher bile salt degradation rate indicates a greater ability to hydrolyze bile salts. The results are shown in Table 2.

[0066] Bile salt degradation rate = (initial bile salt content - bile salt content after culture) / initial bile salt content * 100%;

[0067] HPLC detection method for bile salts: chromatographic column: Ultimate AQ-C18 (5 μm, 4.6×250 mm); flow rate: 1.0 mL / min; detection wavelength: 204 nm; column temperature: 45°C; mobile phase A: phosphate buffer, mobile phase B: acetonitrile, A:B = 56:44 (V:V).

[0068] Table 2: Comparison of the ability of each strain to degrade 6 bile salts

[0069] As shown in Table 2, strain A8 can degrade six bile salts simultaneously with significant degradation ability. Strain A8 was named VB301 and was deposited in the General Microbiology Center of China Culture Collection Administration on July 24, 2023. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO.27998.

[0070] The biological characteristics of strain VB301 are as follows: the strain grows well when cultured anaerobically at 37°C on MRS agar medium for 48-72 hours, and the colonies are milky white, opaque, round, and smooth with a diameter of 1-2 mm. The strain is Gram-positive, and the cells are short rod-shaped under a microscope, as shown in Figure 1 , distributed singly (A in Figure 1 ) or in pairs or clusters (B in Figure 1 ), with oval ends and some forked rods, non-motile, and without spores.

[0071] The sequence of strain VB301 is:

[0072] Example 2 Intestinal tolerance assessment

[0073] The activated VB301 bacterial suspension was inoculated into BBL culture medium at a 1% inoculum size, allowed to stand at 37°C, and cultured anaerobically in an anaerobic workstation for 24 h. Nine 1 mL portions of bacterial liquid were centrifuged to collect the bacteria. One portion was mixed with 1 mL of water and counted as a control. The remaining samples were mixed in a pancreatic juice system consisting of 1 mL of water, 1 mL of artificial intestinal fluid (pH 8.0), 1 mL of 0.03% taurine solution, 1 mL of 0.1% taurine solution, and 1 mL of 0.3% taurine solution. Samples were taken at 0 h, 2 h, and 4 h to count the viable bacteria, and the survival rates at 2 h and 4 h were calculated. The results are shown in Table 3.

[0074] Survival rate calculation: Survival rate = B / A*100%, where A is the counting result after mixing with 1 ml of water; B is the counting result after sample treatment.

[0075] Table 3: Survival of Bifidobacterium animalis subsp. lactis VB301 in different culture systems Note: Unit of viable bacteria: 100 million cfu / mL

[0076] The results in Table 3 show that exposure to 0.03% to 0.3% taurine solutions for 4 hours had little effect on the survival of strain VB301. Even after exposure to 0.3% taurine for 4 hours, the survival rate remained at 91%, demonstrating that strain VB301 has a strong tolerance to taurine. Survival in simulated intestinal fluid at pH 8.0 lasted for 2-4 hours, with a survival rate of 63% after 4 hours, indicating that animal Bifidobacterium lactis subsp. VB301 has excellent activity and tolerance to the intestinal environment.

[0077] Example 3: Evaluation of antibacterial ability against pathogenic bacteria

[0078] The antibacterial ability was tested by the double-layer plate culture method. The lower culture medium was BBL medium. 2 μL of animal Bifidobacterium lactis subsp. VB301 bacterial suspension, strain A3 bacterial suspension, and water were spotted on the BBL solid culture medium. After anaerobic culture at 37°C for 1 day, mature single colonies were formed. The upper layer was poured with a culture medium containing indicator bacteria (Staphylococcus aureus ATCC 6538, Escherichia coli 8099, Salmonella paratyphi B CMCC50094, Listeria monocytogenes ATCC19114, Shigella dysenteriae CMCC51252, Enterococcus faecalis ATCC29212, Clostridium perfringens ATCC 13124), 7-10 mL / dish, and the final concentration of indicator bacteria was 10 6 CFU / mL, after solidification, place it under the appropriate growth conditions of each indicator bacteria and culture it for 16-18 hours, observe and record whether there is an inhibition zone around the single colony of VB301 and the size of the inhibition zone to determine whether it has the activity to inhibit the indicator bacteria.

[0079] Table 4: Evaluation of the antibacterial ability of VB301 against 7 pathogenic bacteria (diameter of inhibition zone, mm)

[0080] The results are shown in Table 4. Animal Bifidobacterium lactis subsp. VB301 has antibacterial effects on all seven pathogens, while another strain of Animal Bifidobacterium lactis subsp. A3 has antibacterial activity against only two pathogens, indicating that the strain of the present invention can treat infectious diseases related to the seven pathogens, including urinary tract infection, purulent abdominal infection, sepsis, endocarditis, diarrhea and fever.

[0081] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", 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 representations 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.

[0082] 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.

[0083]

Claims

1. A Bifidobacterium animalis subsp. lactis VB301, characterized in that The deposit number of the Bifidobacterium animalis subsp. lactis VB301 is CGMCC No.27998.

2. A fermentation supernatant, characterized in that: The invention comprises the metabolites of the animal Bifidobacterium lactis subspecies VB301 as claimed in claim 1.

3. A bacterial suspension, characterized in that It comprises the animal Bifidobacterium lactis subspecies VB301 described in claim 1.

4. Use of the animal Bifidobacterium lactis subsp. VB301 according to claim 1, the fermentation supernatant according to claim 2 or the bacterial suspension according to claim 3 in the preparation of a medicament for preventing and / or treating related diseases caused by high cholesterol; Optionally, the high cholesterol-induced related diseases include any one of hypercholesterolemia, cardiovascular disease, arteriosclerosis and peripheral vascular disease; Optionally, the drug is capable of degrading six bile salts, the six bile salts being sodium taurocholate, sodium taurochenodeoxycholate, sodium taurodeoxycholate, sodium glycocholate, sodium glycochenodeoxycholate, sodium glycodeoxycholate.

5. Use of the animal Bifidobacterium lactis subsp. VB301 according to claim 1 or the bacterial suspension according to claim 3 in the preparation of a medicament for a disease related to at least one of Staphylococcus aureus, Escherichia coli, Salmonella paratyphi B, Listeria monocytogenes, Shigella dysenteriae, Enterococcus faecalis, and Clostridium perfringens.

6. A drug, characterized in that The drug comprises at least one of the animal Bifidobacterium lactis subspecies VB301 described in claim 1, the fermentation supernatant described in claim 2, and the bacterial suspension described in claim 3.

7. The drug according to claim 6, characterized in that The medicament further contains excipients and / or carriers.

8. The drug according to claim 7, 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.

9. The drug according to claim 7, characterized in that 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 cereal hydrolyzed solids.

10. The drug according to claim 6, characterized in that The dosage form of the drug includes at least one selected from oral liquid, powder, granule, capsule, tablet, and pill.

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