CN114381406A

CN114381406A - Bifidobacterium breve CCFM1217 capable of simultaneously reducing blood plasma and caecum trimethylamine and application thereof

Info

Publication number: CN114381406A
Application number: CN202210098006.1A
Authority: CN
Inventors: 王刚; 王茜茜; 赵建新; 陈卫; 张灏
Original assignee: Jiangnan University
Current assignee: Jiangnan University
Priority date: 2022-01-27
Filing date: 2022-01-27
Publication date: 2022-04-22
Anticipated expiration: 2042-01-27
Also published as: CN114381406B

Abstract

The invention discloses bifidobacterium breve CCFM1217 capable of simultaneously reducing blood plasma and caecum trimethylamine and application thereof, belonging to the technical field of microorganisms. The bifidobacterium breve CCFM1217 can reduce the levels of plasma TMAO, plasma TMA and caecum TMA; can improve the structure of intestinal flora, recover intestinal flora disorder caused by high choline, increase the abundance of beneficial bacteria (Roseburia, Rikenella RC9 gut group), reduce the relative abundance of harmful bacteria Anaeroplasma, and reduce the risk of intestinal stress syndrome, obesity, allergy, neurological disease, type II diabetes, non-alcoholic fatty liver disease, and cardiovascular disease. Therefore, the method has wide application value.

Description

Bifidobacterium breve CCFM1217 capable of simultaneously reducing blood plasma and caecum trimethylamine and application thereof

Technical Field

The invention relates to bifidobacterium breve CCFM1217 capable of reducing blood plasma and caecum trimethylamine simultaneously and application thereof, belonging to the technical field of microorganisms.

Background

Cardiovascular disease is the leading cause of morbidity and mortality worldwide, and Atherosclerosis (AS) is the pathological basis of cardiovascular disease. Atherosclerosis is a chronic inflammatory disease involving monocyte and macrophage accumulation, smooth muscle cell proliferation and migration, fibroblast proliferation, cholesterol crystallization and free cholesterol and connective tissue deposition at the lesion. Currently, the accepted initiating factors of AS are arterial wall endothelial damage and lipid deposition; the risk factors include hypertension, blood lipid increase, inflammation, oxidized choline, obesity, smoking, etc.

The intestinal tract of human body contains more than 1000 kinds of microorganisms, and the total number is about 10¹⁴～10¹⁵The mass of the product can reach 1-1.5 kg. The total number of genes encoded by these gut microorganisms is about 100 times the total number of genes encoded by human beings themselves, and thus gut microorganisms are considered to be the second genome of the human body. Intestinal microbial genome and human bodyTogether, the genome, through interaction with environmental factors, affects many important physiological functions of the host, such as food digestion and metabolism, immune response and inflammation, neural activity, etc. The interaction between the intestinal flora and its metabolites with the host is essential to maintain the health of the host. Disorders of intestinal microecology are associated with a number of diseases, including diabetes, obesity, inflammatory bowel disease, neurodegenerative diseases and tumors, among others.

Research shows that intestinal microorganisms mainly affect atherosclerosis through three ways of bacterial infection, cholesterol and lipid metabolism regulation and food and microbial metabolites. TMAO generated by diet-intestinal microorganism-liver-trimethylamine oxide (TMAO) can promote cardiovascular diseases. The microorganism-dependent Trimethylamine (TMA)/TMAO pathway has been shown to be involved in the pathogenesis of cardiovascular disease and is an important diagnostic and therapeutic target for cardiovascular disease. Intervening in the metabolism of intestinal flora may become one of the methods for preventing and treating cardiovascular diseases.

Probiotics have found wide acceptance by consumers as dietary supplements. The supplementary probiotics can directly inject a large amount of beneficial flora into the intestinal tract of a human body, and help to improve the metabolic function of the flora. A large number of scientific researches and clinical experiments prove that the probiotics have obvious improvement effects on constipation, enteritis, lactose intolerance, infection resistance, inflammation, allergy and glycolipid metabolic disorder.

Disclosure of Invention

The invention provides Bifidobacterium breve (Bifidobacterium breve) CCFM1217 which is preserved in Guangdong province microbial strain preservation center at 31.12.12.2021, the preservation address is Guangzhou Miyao No. 100 college 59 and Guangdong province microbial research institute of 5 building, the preservation number is GDMCC No: 62176.

the invention also provides a probiotic preparation containing the bifidobacterium breve CCFM 1217.

In one embodiment, the probiotic formulation has a content of bifidobacterium breve CCFM1217 of ≥ 1 × 10⁹CFU/mL or more than or equal to 1X 10⁹CFU/g。

The invention also provides a composition containing the bifidobacterium breve CCFM 1217.

In one embodiment, the composition further comprises a statin.

The invention also provides a fermented food which is prepared by fermenting and producing the bifidobacterium breve CCFM1217, and the fermented food comprises solid food, liquid food and semi-solid food.

In one embodiment, the fermented food product comprises dairy products, bean products, fruit and vegetable products, and the dairy products comprise milk, sour cream and cheese; the fruit and vegetable products comprise cucumber, carrot, beet, celery and cabbage products.

The invention also provides application of the bifidobacterium breve CCFM1217 in preparing probiotics for in-vivo field planting.

The invention also provides the application of the bifidobacterium breve CCFM1217 in preparing a medicament for reducing the morbidity risk of at least one of intestinal stress syndrome, obesity, allergy, neurological disease, type II diabetes, non-alcoholic fatty liver and cardiovascular disease or a health-care product for relieving at least one disease symptom of the intestinal stress syndrome, obesity, allergy, neurological disease, type II diabetes, non-alcoholic fatty liver and cardiovascular disease; such cardiovascular diseases include, but are not limited to, atherosclerosis.

In one embodiment, the medicament further comprises a pharmaceutically acceptable carrier; the pharmaceutically acceptable carriers include, but are not limited to: one or more of a filler, a wetting agent, a disintegrant, a binder, or a lubricant.

In one embodiment, the use comprises at least one of the following actions:

(1) reducing the level of plasma TMAO;

(2) reducing plasma TMA levels;

(3) reducing the level of cecal TMA;

(4) improve the structure of intestinal flora and increase the abundance of beneficial bacteria.

In one embodiment, the beneficial bacteria comprise genus Roseburia and/or genus rikennellaceae RC9 gut group.

The invention also provides the application of the bifidobacterium breve CCFM1217 in promoting the drug synergism when being used together with the drug.

In one embodiment, the drug includes, but is not limited to, a statin.

The invention has the beneficial effects that: the bifidobacterium breve CCFM1217 provided by the invention can be used for relieving the morbidity risk of atherosclerosis, intestinal stress syndrome, obesity, allergy, neurological diseases, type II diabetes, non-alcoholic fatty liver, cardiovascular diseases and the like, and has a very wide application prospect in the aspect of preparing functional foods, health-care products or medicines. In a high-choline model mouse experiment, the bifidobacterium breve CCFM1217 screened by the invention can be taken to obviously reduce the levels of plasma TMAO, plasma TMA and caecum TMA of the high-choline model mouse, improve the structure of intestinal flora, recover intestinal flora disorder caused by high-choline, improve the abundance of beneficial bacteria (Roseburia, Rikenella asiatica RC9 gut group), reduce the relative abundance of harmful bacteria Anaeroplasma, and reduce the risks of intestinal stress syndrome, obesity, allergy, neuropathy, type II diabetes, nonalcoholic fatty liver and cardiovascular disease.

Biological material preservation

Bifidobacterium breve (Bifidobacterium breve) CCFM1217, classified and named Bifidobacterium breve, has been deposited in Guangdong province microorganism strain collection center at 31.12.2021, with the deposit address of Guangzhou city Michelle No. 100 college No. 59 building, Guangdong province microorganism research institute, and the deposit number is GDMCC No: 62176.

drawings

FIG. 1 shows the colony morphology of Lactobacillus brevis CCFM1217 for fermentation;

FIG. 2 is the effect of Bifidobacterium breve CCFM1217 on plasma TMAO in choline-fed mice; wherein P <0.001, P < 0.0001.

FIG. 3 is the effect of Bifidobacterium breve CCFM1217 on plasma TMA in choline-fed mice; wherein P <0.01, P < 0.001.

FIG. 4 is the effect of Bifidobacterium breve CCFM1217 on caecal TMA in chohne fed mice; wherein P <0.01, P < 0.0001.

FIG. 5 is a graph of the effect of Bifidobacterium breve CCFM1217 on the caecum Roseburia and Rikenellaceae RC9 gut group genera of choline-fed mice; wherein P <0.05, P <0.001, P < 0.0001.

FIG. 6 is a graph showing the effect of Bifidobacterium breve CCFM1217 on the caecum Anaeroplasma genus of choline-fed mice; wherein P < 0.01.

FIG. 7 is a graph of the effect of different Bifidobacterium breve on plasma TMAO in choline-fed mice; wherein P <0.001, P < 0.0001.

Detailed Description

The examples relate to bifidobacterium breve CCFM1217 having the following biological properties:

(1) the characteristics of the thallus are as follows: is gram-positive, does not form a brood, does not move.

(2) Colony characteristics: the bacterial colony is milky white, round, neat in edge, slightly convex, non-transparent and moist and smooth in surface;

(3) growth characteristics: the optimal growth temperature of the strain is 35-37 ℃, the optimal growth pH is 6.5, and the strain enters a stationary phase after being cultured for 18 h;

(4) the level of plasma TMAO can be obviously reduced in a high-choline mouse model;

(5) the level of plasma TMA can be significantly reduced in a high choline mouse model;

(6) the level of cecal TMA can be significantly reduced in a high choline mouse model;

(7) the abundance of Rosebularia and Rikenellaceae RC9 gut group can be obviously improved in a high-choline mouse model, and the risks of intestinal stress syndrome, obesity, allergy, neurological diseases and type II diabetes are reduced;

(8) the abundance of Anaeroplasma can be obviously reduced in a high-choline mouse model, and the risks of obesity, non-alcoholic fatty liver and cardiovascular diseases are reduced.

The extraction method of the bifidobacterium breve CCFM1217 comprises the following steps:

(I) separating and screening the bifidobacterium breve:

(l) 1g of fresh faeces of a healthy person was taken. After gradient dilution, coating the solution on an mMRS solid culture medium, and culturing the medium for 72 hours at 37 ℃ in an anaerobic environment;

(2) observing and recording the colony morphology, selecting colonies, and streaking and purifying;

(3) the colonies were gram-stained in MRS liquid medium at 37 ℃ for 48 hours, and the morphology of the colonies was recorded.

(4) Removing gram-negative bacteria strains and gram-positive cocci from the colonies, and selecting to obtain gram-positive bacilli.

(5) After catalase analysis, catalase-positive strains were discarded, and catalase-negative strains were retained.

(II) molecular biological identification of Bifidobacterium breve:

(l) Extracting a single-bacterium genome: culturing the pediococcus acidilactici obtained by screening in the step (II) overnight, taking the overnight-cultured bacterium suspension lmL to a 1.5mL centrifuge tube, centrifuging for 2min at 10000rpm, and removing the supernatant to obtain thalli; purging the thallus with lmL sterile water, centrifuging at 10000rpm for 2min, and removing the supernatant to obtain thallus; adding 200 μ L SDS lysate, and water bathing at 80 deg.C for 30 min; adding 200 μ L of phenol-chloroform solution into the thallus lysate, wherein the phenol-chloroform solution comprises Tris saturated phenol, chloroform and isoamylol at a volume ratio of 25:24:1, mixing, centrifuging at 12000rpm for 5-10min, and collecting 200 μ L of supernatant; adding 400 μ L of glacial ethanol or glacial isopropanol into 200uL of supernatant, standing at-20 deg.C for 1h, centrifuging at 12000rpm for 5-10min, and discarding the supernatant; adding 500 μ L70% (volume percentage) of glacial ethanol, resuspending the precipitate, centrifuging at 12000rpm for 1-3min, and discarding the supernatant; oven drying at 60 deg.C, or naturally air drying; 50 μ L ddH₂Re-dissolving the precipitate with O for PCR;

(2)16S rDNA PCR：

A. bacterial 16S rDNA 50 μ LPCR reaction:

10 × Taq buffer, 5 μ L; dNTP, 5. mu.L; 27F, 0.5 μ L; 1492R, 0.5 μ L; taq enzyme, 0.5. mu.L; template, 0.5 μ L; ddH₂O，38μL。

PCR conditions:

95℃5min；95℃10s；55℃30s；72℃30s；step2-4 30×；72℃5min；12℃2min；

C. preparing 1% agarose gel, mixing the PCR product with 10000 × loading buffer, loading 2 μ L, running at 120V for 30min, and performing gel imaging;

D. and sending the obtained PCR product to a professional sequencing company, and performing search and similarity comparison on the obtained sequencing result and a GenBank by using BLAST to identify the PCR product as the bifidobacterium breve.

(3) Whole genome sequencing

Sending the extracted whole genome to a professional sequencing company, sequencing the whole genome of the strain by using a second-generation sequencer, searching and comparing similarity of the obtained sequence result in GenBank by using BLAST, and identifying the sequencing result as a newly discovered strain belonging to the bifidobacterium breve. The strain is preserved at-80 ℃ for later use.

Example 1: tolerance of Bifidobacterium breve CCFM1217 to simulated gastrointestinal fluids

Inoculating the cryopreserved bifidobacterium breve CCFM1217 into an MRS culture medium, carrying out anaerobic culture at 37 ℃ for 48h, carrying out subculture for 2-3 times by using the MRS culture medium, mixing 1mL of the culture medium of the bifidobacterium breve CCFM1217 with 9.0mL of artificial simulated gastric juice (MRS culture medium containing 1% pepsin and having a pH value of 2.5), carrying out anaerobic culture at 37 ℃, sampling at 0h, 0.5h, 1h and 2h respectively, carrying out pouring culture by using the MRS agar culture medium, carrying out plate colony counting, measuring the viable count and calculating the survival rate.

The survival rate is the ratio of the logarithmic viable count at the sampling time to the logarithmic viable count at the 0h time in the culture solution, and is expressed by%. Adding 1mL of Bifidobacterium breve CCFM1217 culture solution into 9mL of artificial simulated intestinal fluid (MRS culture medium containing 0.3% of bovine bile salt, 1% of trypsin and pH 8.0), anaerobically culturing at 37 deg.C, sampling at 0h, 0.5h, 1h, 2h, 3h and 4h, pouring and culturing with MRS agar culture medium, counting bacterial colony of plate, measuring viable count and calculating survival rate. The survival rate is the ratio of the logarithmic viable count at the sampling time to the logarithmic viable count at the 0h time in the culture solution, and is expressed by%. The results of the experiment are shown in tables 1 and 2. The result shows that the bifidobacterium breve CCFM1217 has better tolerance to the artificial gastrointestinal fluids.

TABLE 1 tolerance of Bifidobacterium breve CCFM1217 in simulated gastric juice

TABLE 2 tolerance of Bifidobacterium breve CCFM1217 in artificially simulated intestinal fluid

Example 2: bifidobacterium breve CCFM1217 for reducing plasma TMAO levels

24 healthy female C57BL/6J mice of 7 weeks of age, weighing 18-20g, were acclimated for 1 week and randomized into 4 groups: blank Control group (Control), model Control group (Choline), Bifidobacterium breve CCFM1217 group (CCFM1217), Bifidobacterium breve FFJXM1M3 Control group (another Bifidobacterium breve FFJXM1M3 selected from human feces of Ministry of Fujian province was screened according to the same method, the strains are reported in Ruyi Zhu Feng, Hangfeng, Zhanggong, etc.. research on the formation rule and surface property correlation of Bifidobacterium biofilm [ J ]. food and fermentation industry, 2020). Each group of mice had 6 mice, and each mouse was gavaged with 0.2mL of the bacterial solution daily.

The preparation method of the bacterial liquid comprises the following steps: respectively streaking and inoculating bifidobacterium breve CCFM1217 and bifidobacterium breve FFJXM1M3 into an MRS solid culture medium, culturing for 72h at 37 ℃ to obtain single colonies, respectively inoculating the prepared single colonies into an MRS liquid culture medium, and culturing for 24h at 37 ℃ for activation; inoculating the activated 3-generation bacteria liquid into 1L MRS liquid culture medium respectively in an inoculation amount of 2%, shaking, mixing uniformly, and culturing in an anaerobic incubator at 37 ℃ for 24 h. Centrifuging at 8000g/min and 4 deg.C for 15min, removing supernatant, washing with sterile physiological saline (containing 0.05% -0.1% L-cysteine hydrochloride) for 2 times, centrifuging under the same conditions, removing supernatant, and resuspending with 30% glycerol to obtain bacteria concentration not less than 1 × 10⁹CFU/mL of bacterial liquid.

The bacterial liquid is frozen and stored in a refrigerator at the temperature of minus 80 ℃ for one week. Before animal experiment, the refrigerator is usedTaking out the frozen bacteria liquid, centrifuging the bacteria liquid at 6000 r/min for 5min, cleaning twice with sterile normal saline, re-suspending the bacteria liquid with 10% skim milk, shaking uniformly, and measuring the number of the viable bacteria in the initial state and after one week of freezing by using a plate pouring method. The experimental results are as follows: the initial viable count was 3.6X 10⁹CFU/mL，3.0×10⁹CFU/mL, viable count after 1 week was 2.6X 10⁹CFU/mL，2.3×10⁹The magnitude of CFU/mL is not changed, which indicates that the bacteria liquid is frozen and then does not influence the experiment, and the bacteria liquid can be used for animal experiments.

The grouping and treatment methods of the experimental animals are shown in Table 3.

TABLE 3 groups of experimental animals

Week 2-7: the normal group of mice was fed with normal diet, and the remaining mice were fed with choline diet. C57BL/6J mice (female, 7 weeks old) were purchased from Experimental animals technology, Inc. of Weitongli, Beijing. Common feed (LAD 3001M, choline content 0.1%) and choline feed (LAD 3001M, choline content 1.0%) were purchased from Nantong Telofil feed science and technology Co.

Before the experiment was completed, mice were fasted and deprived of water for 4h, and blood was drawn through periorbital capillaries. The blood sample is centrifuged for 15min at 4000 Xg and 4 ℃, the supernatant is taken and frozen in a refrigerator at-80 ℃,20 mu L of the plasma sample is taken and added with 80 mu L (V: V, 1: 4) of acetonitrile to precipitate the protein in the plasma sample, and d9-TMAO with the final concentration of 2.0 mu M is added into the plasma sample as an internal standard. Mixing, standing at-80 deg.C for 2h, at 4 deg.C for 12000g for 15min, sucking supernatant into a sample bottle, storing in-80 deg.C refrigerator, and measuring plasma TMAO level by HPLC-MS/MS.

The results of the plasma TMAO experiment are shown in fig. 2, the plasma TMAO of the choline-fed mice is significantly higher than that of the control mice, and is approximately 5.78 times higher than that of the control mice, compared with the choline-fed mice, the intragastric bifidobacterium breve CCFM1217 significantly reduces the level of the plasma TMAO, compared with the choline-fed mice, the plasma TMAO of the intragastric bifidobacterium breve CCFM1217 mice is reduced by 29.22%, and the plasma TMAO of the intragastric control FFJXM1M3 mice is reduced by only 7.10%.

Example 3: bifidobacterium breve CCFM1217 for reducing plasma TMA levels

The grouping, modeling and handling methods of the C57BL/6J mice were the same as in example 2.

Before the experiment was completed, mice were fasted and deprived of water for 4h, and blood was drawn through periorbital capillaries. Centrifuging blood sample at 4000 Xg and 4 deg.C for 15min, collecting supernatant, and freezing and storing in a refrigerator at-80 deg.C. mu.L of acetonitrile (V: V, 1: 4) was added to 20. mu.L of the plasma sample to precipitate the protein in the plasma sample, and d9-TMAO was added to the plasma sample at a final concentration of 2.0. mu.M as an internal standard. Mixing, standing at-80 deg.C for 2h, at 4 deg.C for 12000g for 15min, sucking supernatant into a sample bottle, storing in a refrigerator at-80 deg.C, and measuring plasma TMA level by HPLC-MS/MS.

The plasma TMA experimental results are shown in fig. 3, the plasma TMA of the mice fed with choline group is significantly higher than that of the mice fed with control group, the plasma TMA of the mice fed with gastric lavage bifidobacterium breve CCFM1217 is significantly reduced by 70.50% compared with the mice fed with choline group, while the plasma TMA of the mice fed with gastric lavage control bacteria FFJXM1M3 is reduced by only 8.05%.

Example 4: bifidobacterium breve CCFM1217 for reducing cecal TMA level

At the end of the test, the mice are fasted and forbidden to drink for 12h, after the mice are anesthetized by intraperitoneal injection of 10% chloral hydrate solution, the ceca of the mice are taken and frozen in a refrigerator at-80 ℃, the ceca content of the mice is weighed, 20 mu L of mixed solution (acetonitrile: methanol: water are mixed according to the volume ratio of 40:40: 20) is added into each mg of ceca content, d9-TMA with the final concentration of 2.5 mu M is added into the ceca sample as an internal standard, the mixture is shaken and mixed evenly, the mixture is kept at-80 ℃ for 2h, 12000g15 min at 4 ℃, supernatant is sucked into a sample bottle and stored in the refrigerator at-80 ℃, and the content of TMA in the ceca of the mice is measured by HPLC-MS/MS.

The results of the cecum TMA experiment are shown in fig. 4, the cecum TMA of the choline-fed mice is significantly higher than that of the control mice, and is about 3.13 times higher than that of the control mice, the level of cecum TMA of the gavage bifidobacterium breve CCFM1217 is significantly reduced compared with that of the choline-fed mice, and the cecum TMA of the gavage bifidobacterium breve CCFM1217 is reduced by 45.14% compared with that of the choline-fed mice, while the cecum TMA of the gavage control bacteria FFJXM1M3 is not reduced, but increased by 12.53%.

Example 5: bifidobacterium breve CCFM1217 for increasing the abundance of beneficial microorganisms in the caecum

After the experiment is finished, the mice are fasted and are forbidden to be watered for 12 hours, 10% chloral hydrate solution is injected into the abdominal cavity for anesthesia, the caecum is taken, caecum DNA is extracted according to the method of the excrement DNA kit, and a second-generation sequencer is used for carrying out 16S rDNA flora analysis on a V3-V4 area of the caecum.

The results of the experiment are shown in FIG. 5. The relative abundance of cecum Roseburia and Rikenella grass RC9 gut group in the choline forage group mice is significantly lower than that of the control forage group, and compared with the choline group mice, the bifidobacterium breve CCFM1217 significantly improves the relative abundance of Roseburia and Rikenella grass RC9 gut group.

Example 6: bifidobacterium breve CCFM1217 for reducing abundance of Anaeroplasma cecum

The results of the experiment are shown in FIG. 6. The relative abundance of the caecum Anaeroplasma genus of the choline forage group mouse is obviously higher than that of the control forage group, and compared with the choline group mouse, the bifidobacterium breve CCFM1217 obviously reduces the relative abundance of the caecum Anaeroplasma genus.

Example 7: comparison of the Effect of different Bifidobacterium breve on plasma TMAO in Choline-fed mice

The mouse modeling and treatment method is the same as that in example 2, 16 different strains of bifidobacterium breve are respectively used for perfusing a mouse fed with choline feed, and plasma TMAO is detected, as shown in figure 7, only CCFM1217 has the capacity of obviously reducing the plasma TMAO of the mouse fed with the choline feed, and the capacity is reduced by 29.22%, and other bifidobacterium breve does not obviously reduce the plasma TMAO of the mouse fed with the choline feed.

Example 8: fermented food containing Bifidobacterium breve CCFM1217 prepared from Bifidobacterium breve CCFM1217

Selecting fresh apples, washing, juicing, carrying out high-temperature instantaneous sterilization, carrying out high-temperature heat sterilization at the temperature of 140 ℃ for 2 seconds, immediately cooling to 37 ℃, and inoculating the bifidobacterium breve CCFM1217 microbial inoculum leavening agent prepared by the invention to ensure that the concentration of the bifidobacterium breve CCFM1217 microbial inoculum leavening agent reaches 10⁸More than CFU/mL, and refrigerating and storing at the temperature of 4 ℃ to obtain the fruit and vegetable beverage containing the viable bacteria of the bifidobacterium breve CCFM 1217.

The bifidobacterium breve CCFM1217 is fermented to produce other fermented foods, wherein the fermented foods comprise solid foods, liquid foods and semi-solid foods. The fermented food comprises dairy products, bean products and fruit and vegetable products, wherein the dairy products comprise milk, sour cream and cheese; the fruit and vegetable products comprise cucumber, carrot, beet, celery and cabbage products.

The prepared fermented food was used to feed choline model mice according to the method of example 2, and the results showed that the fermented food was capable of reducing the levels of plasma TMAO, plasma TMA, caecal TMA; can improve the structure of intestinal flora, recover intestinal flora disorder caused by high choline, increase the abundance of beneficial bacteria (Roseburia, Rikenella RC9 gut group), reduce the relative abundance of harmful bacteria Anaeroplasma, and reduce the risk of intestinal stress syndrome, obesity, allergy, neurological disease, type II diabetes, non-alcoholic fatty liver disease, and cardiovascular disease.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Bifidobacterium breve (CCFM1217) has been deposited at the Guangdong province microorganism culture collection at 31.12.2021, with the deposit address of Guangzhou city Michelia Tokyo 100 # 59 th building 5 th Guangdong province microorganism research institute, with the deposit number being GDMCC No: 62176.

2. probiotic formulation comprising bifidobacterium breve CCFM1217 according to claim 1.

3. The probiotic preparation according to claim 2, characterized in that the content of Bifidobacterium breve CCFM1217 in the probiotic preparation is not less than 1 x 10⁹CFU/mL or more than or equal to 1X 10⁹CFU/g。

4. A pharmaceutical composition comprising Bifidobacterium breve CCFM1217 as claimed in claim 1.

5. The pharmaceutical composition according to claim 4, wherein a statin is included in the composition, but not limited to.

6. A fermented food, characterized in that the fermented food is produced by fermentation using Bifidobacterium breve CCFM 1217.

7. The fermented food product according to claim 6, wherein the fermented food product comprises a dairy product, a soy product, or a fruit and vegetable product.

8. Use of bifidobacterium breve CCFM1217 as claimed in claim 1 for the manufacture of a medicament for reducing the risk of at least one of atherosclerosis, irritable bowel syndrome, obesity, allergy, neurological disease, type II diabetes, non-alcoholic fatty liver disease, cardiovascular disease, or a health product for alleviating at least one of atherosclerosis, irritable bowel syndrome, obesity, allergy, neurological disease, type II diabetes, non-alcoholic fatty liver disease, cardiovascular disease.

9. The use of claim 8, wherein the medicament further comprises a pharmaceutically acceptable carrier.

10. Use of bifidobacterium breve CCFM1217 as claimed in claim 1 to promote drug potentiation when used in combination with a drug.