Animal Bifidobacterium lactis subspecies VB301 and application thereof

Abstract

The present invention discloses a Bifidobacterium animalis subsp. lactis VB301 and an application thereof. The strain was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee on July 24, 2023, with a deposit number of CGMCC No. 27998. The Bifidobacterium animalis subsp. lactis VB301 of the present invention has a significant bile salt lowering ability, can be used to prepare a cholesterol-lowering drug, and has strong intestinal tolerance and antibacterial ability, and has great application prospects in the preparation of medicines.

Description

Technical Field

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

[0002] Cholesterol is an important lipid substance in the human body. At a certain concentration, cholesterol can provide necessary functions for the human body. However, if it is taken in excessive amounts, cholesterol will accumulate on the blood vessel walls to form cholesterol plaques, which in turn cause a variety of cardiovascular diseases. Currently, cardiovascular and cerebrovascular diseases are the main killers of people's deaths, and the trend is increasing year by year. Therefore, lowering the cholesterol level in the body has become an important means of preventing cardiovascular diseases.

[0003] In the liver, the main route for cholesterol is to be converted into bile acid. Glycocholic acid and taurocholic acid derived from bile acid are the main bile acids in humans. Bile acid provides an important excretion pathway for cholesterol metabolism. One-third of cholesterol catabolism is achieved through bile acid synthesis. Bile acids can be divided into two categories: one is free bile acids such as cholic acid, deoxycholic acid, chenodeoxycholic acid and lithocholic acid; the other is conjugated bile acids formed by the combination of free bile acids with glycine or taurine. Among them, there are 6 main types of conjugated bile acids in the human body, including glycochenodeoxycholic acid (26%), glycocholic acid (26%), glycodeoxycholic acid (22%), taurocholic acid (9%), taurodeoxycholic acid (9%) and taurochenodeoxycholic acid (9%). Bile acids exist in the form of sodium or potassium salts, namely bile salts, referred to as bile salts.

[0004] The mechanisms proposed for probiotics to lower 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 co-precipitate in an acidic environment, thereby lowering serum cholesterol levels; (3) probiotics ferment carbohydrates from intestinal food sources 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 six major bile salts bound to the human body, the six bile salts being 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 prominent. Summary of the invention

[0006] The present invention aims to solve at least one of the technical problems existing in the prior art to at least a certain extent.

[0007] To this end, the first aspect of the present invention provides a Bifidobacterium animalis subsp. lactis VB301, wherein the Bifidobacterium animalis subsp. lactis VB301 was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration on July 24, 2023, with a deposit number of CGMCC No. 27998.

[0008] The animal Bifidobacterium lactis subspecies provided by the present invention has the ability to simultaneously degrade 6 types of bound bile salts, intestinal tolerance and antibacterial ability, and can be used to prevent and / or alleviate and / or treat related diseases caused by high cholesterol on the one hand, and 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, suppurative abdominal infection, sepsis, endocarditis, diarrhea and fever, etc.

[0009] The second aspect of the present invention provides a fermentation supernatant, wherein the fermentation supernatant comprises the metabolites of the animal Bifidobacterium lactis subspecies VB301 described in the first aspect.

[0010] The third aspect of the present invention provides a bacterial suspension, which includes the animal Bifidobacterium lactis subspecies VB301 described in the first aspect.

[0011] 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 related diseases caused by high cholesterol.

[0012] 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.

[0013] 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.

[0014] 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 a disease related to Staphylococcus aureus, Escherichia coli, Salmonella paratyphi B, Listeria monocytogenes, Shigella dysenteriae, Enterococcus faecalis, and Clostridium perfringens.

[0015] 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, etc.

[0016] The sixth aspect of the present invention provides a medicine, which comprises at least one of the animal Bifidobacterium lactis subspecies VB301 described in the first aspect, the fermentation supernatant described in the second aspect, and the bacterial suspension described in the third aspect.

[0017] According to an embodiment of the present invention, the medicament further contains an excipient and / or a carrier.

[0018] 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.

[0019] 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.

[0020] 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.

[0021] According to an embodiment of the present invention, when animal Bifidobacterium lactis subspecies VB301 exerts its effect, it may exist in the form of living cells or in the form of non-living cells.

[0022] 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).

[0023] In addition, the present invention provides a microbial agent, which contains the aforementioned animal Bifidobacterium lactis subspecies 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.

[0024] The present invention also provides the use of the aforementioned animal Bifidobacterium lactis subspecies VB301 and / or the aforementioned fermentation supernatant in the preparation of health products and foods, wherein the health products or foods are used to prevent and / or treat related diseases caused by high cholesterol.

[0025] 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.

[0026] Collection information:

[0027] Strain name: VB301

[0028] Deposit date: July 24, 2023

[0029] Depository: China General Microbiological Culture Collection Center (CGMCC)

[0030] Deposit number: CGMCC No.27998 BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The microscopic morphology of Bifidobacterium animalis subsp. lactis in Example 1 of the present invention is shown, wherein: Figure 1 A in the figure indicates the morphology of a single animal Bifidobacterium lactis subspecies VB301 under a microscope. Figure 1 B in the figure indicates the morphology of clustered Bifidobacterium animalis subsp. lactis VB301 under a microscope. DETAILED DESCRIPTION

[0032] Embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to be used to explain the present invention, but should not be understood as limiting the present invention.

[0033] 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.

[0034] In this document, the terms “contain”, “include” or “comprise” are open expressions, that is, they include the contents specified in the present invention but do not exclude other contents.

[0035] 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.

[0036] As used herein, "prevention" and "prevention" are used interchangeably. These terms refer to an approach to obtaining a beneficial or desired result, including but not limited to a prophylactic benefit. To obtain a "prophylactic benefit," fermented Lactobacillus muciniphila or a product containing it may be administered to a subject at risk of 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 been made.

[0037] 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.

[0038] As used herein, "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.

[0039] In this article, "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 mammals treated therewith. Preferably, "pharmaceutically acceptable" as described in the present invention refers to those approved by federal regulatory agencies or national governments or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeias for use in animals, particularly humans.

[0040] As used herein, the term "pharmaceutically acceptable 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 encompassed.

[0041] In this article, the term "pharmaceutically acceptable excipients" may include carbohydrates, 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 and other excipients commonly used in pharmaceutical preparations.

[0042] According to a specific embodiment of the present invention, the present invention proposes a Bifidobacterium animalis subsp. lactis VB301. According to an embodiment of the present invention, the preservation number of the Bifidobacterium animalis subsp. lactis is CGMCC No. 27998. The Bifidobacterium animalis subsp. lactis of the present invention has a significant bile salt lowering ability, and the Bifidobacterium animalis subsp. lactis VB301 has strong intestinal tolerance and antibacterial ability.

[0043] As used herein, "Bifidobacterium animalis subsp. lactis VB301" and "Bifidobacterium animalis subsp. lactis VB301", "VB301", "strain VB301" are synonymous.

[0044] The animal Bifidobacterium lactis subspecies VB301 obtained by screening according to the present invention has a significant bile salt degradation ability compared with other animal Bifidobacterium lactis subspecies. The strain VB301 according to the present invention can degrade 6 bile salts, which are sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), sodium glycocholate (GC), sodium glycochenodeoxycholate (GCDC), and sodium glycodeoxycholate (GDC).

[0045] The animal bifidobacterium lactis subspecies (Bifidobacterium animalis subsp.lactis) VB301 of the present invention has stronger intestinal tolerance than other animal bifidobacterium lactis subspecies. The strain VB301 of the present invention has substantially no effect on bacterial survival in a 0.03% to 0.3% oxcholine solution for 4 hours, and the survival rate can still reach 91% in a 0.3% oxcholine solution for 4 hours, indicating that the strain VB301 has a strong tolerance to oxcholine. It can survive in a simulated intestinal fluid of pH 8.0 for 2-4 hours, and the 4-hour survival rate can still reach 63%, indicating that the animal bifidobacterium lactis subspecies VB301 has a good active tolerance to the intestinal environment and can play a role in the intestinal tract for a long time.

[0046] The animal Bifidobacterium lactis subspecies VB301 of the present invention has stronger antibacterial ability than other animal Bifidobacterium lactis subspecies, and the animal Bifidobacterium lactis subspecies VB301 has antibacterial effect on 7 pathogenic bacteria, wherein the 7 pathogenic bacteria are Escherichia coli, Staphylococcus aureus, Salmonella, Listeria, Shigella, Enterococcus faecalis and Clostridium perfringens.

[0047] According to a specific embodiment of the present invention, the present invention provides a fermentation supernatant, the fermentation supernatant comprising: animal Bifidobacterium lactis subspecies VB301 and / or metabolites of animal Bifidobacterium lactis subspecies VB301. The animal Bifidobacterium lactis subspecies VB301 of the present invention has a significant bile salt lowering ability, and the animal Bifidobacterium lactis subspecies VB301 has strong intestinal tolerance and antibacterial ability.

[0048] It should be noted that "fermentation supernatant" refers to the supernatant after centrifugation of the fermentation broth, which may contain animal Bifidobacterium lactis subsp. VB301 or metabolites of animal Bifidobacterium lactis subsp. VB301, or both animal Bifidobacterium lactis subsp. VB301 and its metabolites.

[0049] According to a specific embodiment of the present invention, the present invention provides a microbial agent, wherein the microbial agent contains the aforementioned animal Bifidobacterium lactis subspecies VB301 and / or the aforementioned fermentation supernatant. The microbial agent of the present invention is used for preventing and / or treating related diseases caused by high cholesterol.

[0050] 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 may be a solid microbial agent, including but not limited to freeze-dried powder, etc.

[0051] According to an embodiment of the present invention, the Bifidobacterium animalis subspecies lactis VB301 exists in the form of living cells and / or non-living cells.

[0052] As used herein, "living cells" refer to cells that have the ability to metabolize, reproduce or replicate.

[0053] Exemplarily, the living cells may be immobilized cells. Herein, "immobilized cells" refer to Bifidobacterium animalis subsp. lactis VB301 fixed on a carrier, which can carry out life activities such as growth, development, reproduction, inheritance and metabolism within a certain spatial range.

[0054] Herein, "non-living 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.

[0055] As a specific embodiment, the Bifidobacterium animalis subspecies lactis VB301 exists in the form of living cells, dry bacteria, immobilized cells or any other forms.

[0056] As a specific embodiment, the dry bacteria are obtained by freeze-drying the animal Bifidobacterium lactis subspecies VB301.

[0057] According to an embodiment of the present invention, the present invention provides a medicine or health product or food, which contains animal Bifidobacterium lactis subspecies VB301. Preferably, the medicine or health product or food of the present application is a live bacterial preparation for lowering cholesterol. 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 combination with other active ingredients, as long as the activity is not affected mutually; further, the best situation of the medicine or health care product or food is that the active ingredients can have functional complementarity or have a promoting effect, for example, animal Bifidobacterium lactis subspecies VB301 can be combined with other probiotics to form a composite probiotic tablet, which can be used to prevent and / or alleviate and / or treat related diseases caused by high cholesterol on the one hand, and 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, Clostridium perfringens, including infectious diseases, specifically, it 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, and an acceptable carrier or excipient in health food. It should be noted that when preparing live bacteria preparations, carriers or excipients are usually required to be added, as long as there is no mutual inhibition or adverse side effects between the added carriers or excipients and animal Bifidobacterium lactis subspecies VB301. The medicine or health product or food can be powder, tablet, beverage, capsule.

[0058] 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.

[0059] Example 1 Strain collection and identification

[0060] 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. Then, the suspension was prepared by oscillation and homogenization. 1 mL was taken for gradient dilution to prepare different gradients of diluted bacterial suspensions, which were spread on MRS agar plates containing 50 μg / mL mupirocin and 5% serum (V / V), and cultured anaerobically at 37°C for 48-72 h. 130 colonies with different morphologies were picked, streaked and purified, and DNA was extracted. PCR amplification was performed with specific primers, and 16s rDNA was sequenced. At the same time, 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 finally 18 strains of bifidobacteria were identified and separated, 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.

[0061] Table 1: Strain collection and identification results

[0062]

[0063] The 18 strains were tested for their ability to degrade six bile salts, namely sodium taurocholate (TAC), sodium taurochenodeoxycholate (TCDC), sodium taurodeoxycholate (TDC), sodium glycocholate (GC), sodium glycochenodeoxycholate (GCDC), and sodium glycodeoxycholate (GDC).

[0064] Six different bile salts were added to the BBL liquid medium, and the bile salt concentration was 2-4mmol / L. The activated glycerol tubes of each strain were taken out from the -80℃ refrigerator, and after thawing, they were inoculated into the BBL liquid medium at a 1-3% inoculation rate. After standing at 37℃, anaerobic culture was performed in an anaerobic workstation for 24 hours to obtain the activated bacterial solution, and 1mL of the culture solution was taken, centrifuged, and the supernatant was taken. The bile salt content was detected by HPLC, and the bile salt degradation ability of each strain was evaluated by calculating the bile salt degradation rate. The higher the bile salt degradation rate, the stronger the ability to hydrolyze bile salts. The results are shown in Table 2.

[0065] Bile salt degradation rate = (initial bile salt content - bile salt content after culture) / initial bile salt content * 100%;

[0066] HPLC detection method for bile salts: chromatographic column: Ultimate AQ-C18 (5um, 4.6×250mm); flow rate: 1.0mL / min; detection wavelength: 204nm; column temperature: 45°C; mobile phase A: phosphate buffer, mobile phase B: acetonitrile, A:B=56:44 (V:V).

[0067] Table 2: Comparison of the ability of each strain to degrade 6 bile salts

[0068]

[0069] As can be seen from Table 2, the A8 strain can degrade 6 bile salts at the same time and has a significant degradation ability. The A8 strain was named VB301 and was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on July 24, 2023. The deposit address is No. 3, Yard No. 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 for 48-72 hours on MRS agar medium. The colonies are milky white, opaque, round, and smooth, with a colony diameter of 1-2 mm. The strain is Gram-positive, and the cell morphology under a microscope is short rod-shaped, such as Figure 1 As shown, a single ( Figure 1 A) or in pairs or clusters ( Figure 1 B) in the figure has oval ends, some are forked rod-shaped, non-motile, and without spores.

[0071] The sequence of strain VB301 is:

[0072]

[0073] Example 2 Intestinal tolerance assessment

[0074] The activated VB301 bacterial suspension was inoculated into BBL culture medium at a 1% inoculum size and allowed to stand at 37°C. Anaerobically cultured for 24 h in an anaerobic workstation, 9 portions of 1 mL of bacterial solution were centrifuged to collect the bacteria, one of which was mixed with 1 mL of water and counted as a control, and the remaining samples were mixed in a pancreatic juice system of 1 mL of water, 1 mL of artificial intestinal fluid (pH 8.0), 1 mL of 0.03% oxcholine solution, 1 mL of 0.1% oxcholine solution, and 1 mL of 0.3% oxcholine solution. The live bacteria were counted at 0 h, 2 h, and 4 h, and the survival rates at 2 h and 4 h were calculated. The results are shown in Table 3.

[0075] 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.

[0076] Table 3: Survival of Bifidobacterium animalis subsp. lactis VB301 in different culture systems

[0077]

[0078] Note: Unit of viable bacteria: 100 million cfu / mL

[0079] The results in Table 3 show that the strain VB301 has little effect on the survival of the bacteria when exposed to 0.03% to 0.3% taurine solution for 4 hours, and the survival rate can still reach 91% after 4 hours of exposure to 0.3% taurine, indicating that the strain VB301 has a strong tolerance to taurine. It can survive for 2-4 hours in the simulated intestinal fluid of pH 8.0, and the survival rate can still reach 63% after 4 hours, indicating that animal Bifidobacterium lactis subspecies VB301 has a good active tolerance to the intestinal environment.

[0080] Example 3: Evaluation of antibacterial ability against pathogenic bacteria

[0081] The antibacterial ability was tested by double-layer plate culture method. The lower culture medium was BBL culture medium. 2 μL of animal Bifidobacterium lactis subsp. VB301 bacterial suspension, strain A3 bacterial suspension and water were inoculated on BBL solid culture medium. After anaerobic culture at 37°C for 1 day, mature single colonies were formed. The upper layer was poured with 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 6CFU / mL, after solidification, place it under the appropriate growth conditions for each indicator bacteria and culture it for 16-18h, 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.

[0082] Table 4: Evaluation of the antibacterial ability of VB301 against 7 pathogenic bacteria (diameter of inhibition zone, mm)

[0083]

[0084]

[0085] The results are shown in Table 4. Animal Bifidobacterium lactis subspecies VB301 has an antibacterial effect on all 7 pathogenic bacteria, and another strain of Animal Bifidobacterium lactis subspecies A3 has an antibacterial activity on only 2 pathogenic bacteria, indicating that the strain of the present invention can treat infectious diseases related to the 7 pathogenic bacteria, including urinary tract infection, purulent abdominal infection, sepsis, endocarditis, diarrhea and fever, etc.

[0086] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", 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, without contradiction.

[0087] 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

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

3. Use of the animal Bifidobacterium lactis subspecies VB301 according to claim 1 or the bacterial suspension according to claim 2 in the preparation of a medicament for preventing and / or treating diseases related to high cholesterol; Optionally, the high cholesterol-induced related diseases include any one of hypercholesterolemia and cardiovascular 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.

4. The use according to claim 3, characterized in that: The cardiovascular disease includes any one of arteriosclerosis and peripheral vascular disease.

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 medicine comprises at least one of the animal Bifidobacterium lactis subspecies VB301 described in claim 1 and the bacterial suspension described in claim 2.

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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