CN114381406B

CN114381406B - Bifidobacterium breve CCFM1217 capable of simultaneously reducing blood plasma and cecum trimethylamine and application thereof

Info

Publication number: CN114381406B
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: 2023-06-13
Anticipated expiration: 2042-01-27
Also published as: CN114381406A

Abstract

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

Description

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

Technical Field

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

Background

Cardiovascular disease is the leading cause of morbidity and mortality worldwide, atherosclerosis (AS) being the pathological basis of cardiovascular disease. Atherosclerosis is a chronic inflammatory disease that involves the accumulation of monocytes and macrophages at lesions, smooth muscle cell proliferation and migration, fibroblast proliferation, cholesterol crystallization, and free cholesterol and connective tissue deposition. Currently, the accepted initiating factor for AS is arterial wall endothelial injury and lipid deposition; the dangerous factors are hypertension, elevated blood lipid, inflammation, oxidized high choline, obesity, smoking, etc.

The intestinal tract of human body contains more than 1000 microorganisms, and the total number is about 10 ¹⁴ ～10 ¹⁵ The mass of the powder can reach 1-1.5 kg. The total number of genes encoding these intestinal microorganisms is about 100 times the total number of genes encoding the human self-encoding genes, so that the intestinal microorganisms are considered to be the second genome of the human body. The intestinal microbial genome, together with the human genome, affects a variety of important physiological functions such as food digestion and metabolism, immune response and inflammation, neural activity, etc. of the host through interaction with environmental factors. The interaction between the intestinal flora and its metabolites and the host is essential for maintaining 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, and the like.

Studies have shown that intestinal microorganisms affect atherosclerosis mainly through bacterial infection, regulation of cholesterol and lipid metabolism, food and microbial metabolites. TMAO produced via the dietary-intestinal microbial-liver-trimethylamine oxide (TMAO) pathway may promote the development of cardiovascular disease. The microbial-dependent Trimethylamine (TMA)/TMAO pathway has been shown to be associated with the pathogenesis of cardiovascular disease and is an important diagnostic and therapeutic target for cardiovascular disease. Intervention in the metabolism of the intestinal flora, or possibly one of the methods for preventing and treating cardiovascular diseases.

Probiotics have been widely accepted by consumers as dietary supplements. The supplementing probiotics can directly inject a large amount of beneficial bacteria into human intestinal tracts, so as to help improve the metabolic function of the bacteria. Numerous scientific studies and clinical experiments have demonstrated that probiotics have a significant improving effect on constipation, enteritis, lactose intolerance, anti-infection, inflammation, allergy, and glycolipid metabolic disorders.

Disclosure of Invention

The invention provides a bifidobacterium breve (Bifidobacterium breve) CCFM1217 which is preserved in the microorganism strain collection of Guangdong province at 12 months 31 of 2021, wherein the preservation address is the building 5 building Guangdong province microorganism research institute of No. 59 of the Ming's 100 of Guangzhou City, and the preservation number is GDMCC No:62176.

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

In one embodiment, the content of the bifidobacterium breve CCFM1217 in the probiotic preparation is more than or equal to 1 multiplied by 10 ⁹ CFU/mL or ≡1X10. ⁹ CFU/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 produced by using the bifidobacterium breve CCFM1217 for fermentation production, and the fermented food comprises solid food, liquid food and semi-solid food.

In one embodiment, the fermented food comprises dairy products, soy products, fruit and vegetable products, the dairy products comprising milk, sour cream, 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 in-vivo colonization probiotics.

The invention also provides application of the bifidobacterium breve CCFM1217 in preparing medicaments for reducing the incidence risk of at least one of the diseases of the irritable bowel syndrome, the obesity, the allergy, the nervous diseases, the type II diabetes, the nonalcoholic fatty liver and the cardiovascular diseases; the cardiovascular disease includes, but is 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, wetting agent, disintegrant, binder, or lubricant.

In one embodiment, the application includes at least one of the following actions:

(1) Lowering plasma TMAO levels;

(2) Lowering plasma TMA levels;

(3) Lowering the level of cecal TMA;

(4) Improving the structure of intestinal flora and increasing the abundance of beneficial bacteria.

In one embodiment, the beneficial bacteria include Roseburia and/or rickenella caena RC9 gun group.

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

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 risks of atherosclerosis, irritable bowel syndrome, obesity, allergy, neurological diseases, type II diabetes, nonalcoholic fatty liver, cardiovascular diseases and the like, and has very wide application prospects in the aspect of preparing functional foods, health-care products or medicines. In the experiment of the high choline model mice, the administration of the screened bifidobacterium breve CCFM1217 can obviously reduce the levels of plasma TMAO, plasma TMA and cecum TMA of the high choline model mice, improve the structure of intestinal flora, recover intestinal flora disorder caused by high choline, improve the abundance of beneficial bacteria (Roseburia, rikenella eae RC9 gun group), reduce the relative abundance of harmful bacteria Anaerotoplasta, and reduce the risks of occurrence of irritable bowel syndrome, obesity, allergy, neurological diseases, type II diabetes, nonalcoholic fatty liver and cardiovascular diseases.

Preservation of biological materials

Bifidobacterium breve (Bifidobacterium breve) CCFM1217, classified as Bifidobacterium breve, was deposited at the Guangdong province microbiological bacterial collection center at 12 months 31 of 2021, at accession number GDMCC No:62176.

drawings

FIG. 1 shows the colony morphology of the lactic acid bacterium Bifidobacterium breve 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 of choline-fed mice; wherein P <0.01 and P <0.001.

FIG. 4 is the effect of Bifidobacterium breve CCFM1217 on cecal TMA of choline-fed mice; wherein P <0.01 and P <0.0001.

FIG. 5 is the effect of Bifidobacterium breve CCFM1217 on the cecum Roseburia and Rikenella eae RC9 gun group of choline fed mice; wherein P <0.05, P <0.001, P <0.0001.

FIG. 6 is the effect of Bifidobacterium breve CCFM1217 on the cecum Anaeroplasma genus of choline fed mice; wherein P <0.01.

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

Detailed Description

The bifidobacterium breve CCFM1217 of the examples has the following biological properties:

(1) Characteristics of the cells: gram-positive staining, no formation of embracing and no movement of bacteria.

(2) Colony characteristics: the bacterial colony is milky white, round, regular in edge, microprotrusion, opaque 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 stable period after being cultured for 18 hours;

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

(5) The plasma TMA level 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 the Roseburia genus and the Rikenella eae RC9 gun group genus can be obviously improved in a high choline mouse model, and the risks of the irritable bowel syndrome, obesity, allergy, neurological diseases and type II diabetes are reduced;

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

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

isolation and screening of Bifidobacterium breve:

(l) 1g of fresh feces of healthy people is taken. After gradient dilution, the mixture is coated on an mMRS solid culture medium and is placed in an anaerobic environment for culturing for 72 hours at 37 ℃;

(2) Observing and recording the morphology of the bacterial colony, picking the bacterial colony, and streaking and purifying;

(3) The colonies obtained were gram stained in MRS liquid medium at 37℃for 48 hours, and colony morphology was recorded.

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

(5) After the catalase analysis, the catalase positive strain was discarded, and the catalase negative strain was retained.

(II) molecular biological identification of Bifidobacterium breve:

(l) Single bacterial genome extraction: culturing the pediococcus acidilactici obtained by screening in the step (II) overnight, taking a bacterial suspension lmL cultured overnight, centrifuging at 10000rpm for 2min in a 1.5mL centrifuge tube, and discarding the supernatant to obtain thalli; washing the thalli with lmL sterile water, centrifuging at 10000rpm for 2min, and discarding the supernatant to obtain thalli; 200 mu L of SDS lysate is added, and water bath is carried out for 30min at 80 ℃; 200 mu L of phenol-chloroform solution is added into the bacterial lysate, wherein the phenol-chloroform solution comprises the components and volume ratio of Tris saturated phenol, chloroform and isoamyl alcoholAfter mixing upside down, centrifuging at 12000rpm for 5-10min, and collecting 200 μl of supernatant; adding 400 μl of glacial ethanol or glacial isopropanol into 200uL of supernatant, standing at-20deg.C for 1h, centrifuging at 12000rpm for 5-10min, and discarding supernatant; adding 500 μL70% (volume percent) of ice ethanol to resuspend the precipitate, centrifuging at 12000rpm for 1-3min, and discarding the supernatant; oven drying at 60deg.C, or naturally air drying; 50 mu L ddH ₂ O redissolving and precipitating to prepare PCR;

(2)16S rDNAPCR：

A. bacterial 16s rdna50 μlpcr reaction system:

10×Taq buffer, 5. Mu.L; dNTP, 5. Mu.L; 27F, 0.5. Mu.L; 1492R, 0.5. Mu.L; taq enzyme, 0.5. Mu.L; template, 0.5 μl; ddH ₂ O，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 μl, running at 120V for 30min, and performing gel imaging;

D. the obtained PCR product was sent to a professional sequencing company, and the obtained sequencing result was compared with the search and similarity in GenBank using BLAST, and identified as Bifidobacterium breve.

(3) Whole genome sequencing

The extracted whole genome is sent to a professional sequencing company, a second generation sequencer is used for sequencing the whole genome of the bacterium, the obtained sequence result is searched in a GenBank by using BLAST and is subjected to similarity comparison, and the sequencing result is identified as a newly discovered strain belonging to bifidobacterium breve. The strain is preserved at-80 ℃ for standby.

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

The frozen and preserved bifidobacterium breve CCFM1217 is inoculated in MRS culture medium, anaerobic cultured for 48 hours at 37 ℃, subcultured for 2-3 times by MRS culture solution, 1mL of culture solution of bifidobacterium breve CCFM1217 is taken, mixed with 9.0mL of artificial simulated gastric fluid (MRS culture medium containing 1% pepsin and pH=2.5) with pH of 2.5, anaerobic cultured at 37 ℃, sampled at 0h, 0.5h, 1h and 2h respectively, plate colony count is carried out by casting culture by MRS agar culture medium, viable bacteria number is measured, and survival rate is calculated.

The survival rate is the ratio of the number of viable bacteria to the number of viable bacteria at the time of sampling in the culture solution to the number of viable bacteria at the time of 0h, expressed in%. 1mL of a culture solution of Bifidobacterium breve CCFM1217 was added to 9mL of an artificial simulated intestinal fluid (MRS medium containing 0.3% of bovine bile salt, 1% of trypsin, pH=8.0), anaerobic culture was performed at 37℃and samples were taken at 0h, 0.5h, 1h, 2h, 3h and 4h, respectively, plate colony counts were performed by casting culture with MRS agar medium, the viable count was determined and the survival rate was calculated. The survival rate is the ratio of the number of viable bacteria to the number of viable bacteria at the time of sampling in the culture solution to the number of viable bacteria at the time of 0h, expressed in%. The experimental results are shown in tables 1 and 2. The result shows that the bifidobacterium breve CCFM1217 has better tolerance to the artificial gastrointestinal fluid.

TABLE 1 tolerance of Bifidobacterium breve CCFM1217 in artificial simulated gastric fluid

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

Example 2: bifidobacterium breve CCFM1217 for reducing plasma TMAO levels

24 healthy female C57BL/6J mice with a weight of 18-20g and a age of 7 weeks are taken and are adapted to the environment for 1 week, and randomly divided into 4 groups: blank Control (Control), model Control (Choline), bifidobacterium breve CCFM1217 (CCFM 1217), bifidobacterium breve FFJXM1M3 Control (another strain of bifidobacterium breve FFJXM1M3 selected from human feces of Xiamen, fujian, the same method was screened, strain was reported in Zhu Ruyi, hang Feng, zhang, etc.. Bifidobacterium biofilm formation rule and surface property correlation study [ J ]. Food and fermentation industry, 2020). Each group of mice contains 6 mice, and each mouse is filled with 0.2mL of bacterial liquid every day.

The preparation method of the bacterial liquid comprises the following steps: respectively streaking and inoculating the bifidobacterium breve CCFM1217 and the bifidobacterium breve FFJXM1M3 into an MRS solid culture medium, culturing for 72 hours at 37 ℃ to obtain single colonies, respectively inoculating the prepared single colonies into an MRS liquid culture medium, and culturing for 24 hours at 37 ℃ for activation; the bacterial solutions after 3 generations of activation are respectively inoculated into 1L of MRS liquid culture medium with the inoculation amount of 2 percent, and are cultured for 24 hours at 37 ℃ in an anaerobic incubator after being evenly mixed by shaking. Centrifuging at 8000g/min at 4deg.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 re-suspending with 30% glycerol to obtain strain with concentration of no less than 1×10 ⁹ CFU/mL bacterial liquid.

Freezing the bacterial liquid at-80 deg.c in refrigerator for one week. Before animal experiments are carried out, the frozen bacterial liquid in the refrigerator is taken out, the bacterial liquid is centrifuged for 5min at 6 000r/min, then the bacterial liquid is washed twice with sterile physiological saline, the bacterial liquid is resuspended with 10% skim milk, and after shaking is uniform, the number of viable bacteria after the initial and one week of freezing is measured by a flat plate pouring method. Experimental results: the initial viable count is 3.6X10 respectively ⁹ CFU/mL，3.0×10 ⁹ CFU/mL, viable count after 1 week was 2.6X10 ⁹ CFU/mL，2.3×10 ⁹ The order of CFU/mL is not changed, which indicates that the bacterial liquid can not influence the experiment after being frozen and can be used for animal experiments.

The experimental animal groups and treatment methods are shown in Table 3.

TABLE 3 grouping of experimental animals

Week 2-7: normal mice were fed normal diet and the remaining mice were fed choline diet. C57BL/6J mice (female, 7 week old) were purchased from Peking Vitre Liwa laboratory animal technologies Co. Common feeds (LAD 3001M, choline content 0.1%) and choline feeds (LAD 3001M, choline content 1.0%) were purchased from south-through terlafei feed technologies.

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

Plasma TMAO experimental results as shown in fig. 2, the plasma TMAO of the choline feed group mice was significantly higher than the control feed group, about 5.78 times higher than the control group, the bifidobacterium intragastmeans CCFM1217 significantly reduced the plasma TMAO level compared to the choline group mice, the plasma TMAO of the bifidobacterium intragastmeans CCFM1217 mice was reduced by 29.22% compared to the choline group mice, and the plasma TMAO of the intragastric control bacteria FFJXM1M3 mice was reduced by only 7.10%.

Example 3: bifidobacterium breve CCFM1217 for reducing plasma TMA levels

The grouping, modeling and processing method of the C57BL/6J mice are the same as in example 2.

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

As shown in fig. 3, the plasma TMA of the choline feed group mice was significantly higher than the control feed group, the bifidobacterium intragastricatum CCFM1217 significantly reduced the plasma TMA level compared to the choline group mice, and the plasma TMA of the bifidobacterium intragastricatum CCFM1217 mice was reduced by 70.50% compared to the choline group mice, while the plasma TMA of the intragastricatum FFJXM1M3 mice was reduced by only 8.05%.

Example 4: bifidobacterium breve CCFM1217 for reducing cecum TMA levels

At the end of the test, the mice are fasted and forbidden for 12 hours, 10% chloral hydrate solution is injected into the abdominal cavity for anesthesia, the cecum of the mice is taken and frozen in a refrigerator at-80 ℃,20 mu L of mixed solution (acetonitrile: methanol: water is mixed according to the volume ratio of 40:40:20) is added into each mg of cecum content, d9-TMA with the final concentration of 2.5 mu M is added into a cecum sample as an internal standard, the mixture is uniformly mixed by shaking, the mixture is kept stand at-80 ℃ for 2 hours, 12000g for 15 minutes at 4 ℃, the supernatant is absorbed into a sample bottle, the sample bottle is stored in the refrigerator at-80 ℃, and the TMA content in the cecum of the mice is measured by HPLC-MS/MS.

As shown in fig. 4, the cecum TMA test results show that the cecum TMA of the choline feed group is significantly higher than that of the control feed group, which is about 3.13 times higher than that of the control group, the bifidobacterium intragastrical CCFM1217 significantly reduces the cecum TMA level, and the cecum TMA of the bifidobacterium intragastrical CCFM1217 is reduced by 45.14% compared with that of the choline group, while the cecum TMA of the mice of the intragastrical control bacterium FFJXM1M3 is not reduced, but is increased by 12.53%.

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

At the end of the test, the mice are fasted and forbidden for 12 hours, after the 10% chloral hydrate solution is injected into the abdominal cavity for anesthesia, the cecum is taken, cecum DNA is extracted according to the method of a fecal DNA kit, and the second generation sequencer is used for carrying out 16S rDNA flora analysis on the V3-V4 region of the cecum.

The experimental results are shown in FIG. 5. The relative abundance of the cecum Roseburia and the rikenella eae RC9 gun group was significantly lower in the choline fed group mice than in the control feed group, and bifidobacterium oxydans CCFM1217 significantly increased the relative abundance of the Roseburia and the rikenella eae RC9 gun group compared to the choline group mice.

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

The experimental results are shown in FIG. 6. The relative abundance of Anaeroplasma in the cecum of mice in the choline feed group was significantly higher than that of the control feed group, and the bifidobacterium oxydans CCFM1217 significantly reduced the relative abundance of Anaeroplasma in the cecum compared to the choline group mice.

Example 7: comparing the effect of different bifidobacterium breve on plasma TMAO of choline feed-fed mice

The mice were subjected to the same modeling and treatment as in example 2, and plasma TMAO was detected in mice fed with 16 different strains of bifidobacterium breve gavage choline feed, and only CCFM1217 showed a significant reduction in plasma TMAO in choline feed fed mice, by 29.22% and no significant reduction in TMAO in choline feed fed mice.

Example 8: production of fermented food containing Bifidobacterium breve CCFM1217 by using Bifidobacterium breve CCFM1217

Selecting fresh apples, cleaning, squeezing, performing high-temperature instant sterilization, performing high-temperature sterilization at 140 ℃ for 2 seconds, immediately cooling to 37 ℃, and then inoculating the bifidobacterium breve CCFM1217 microbial inoculum starter prepared by the method to ensure that the concentration reaches 10 ⁸ And (3) refrigerating and preserving the mixture at the temperature of 4 ℃ above CFU/mL to obtain the fruit and vegetable beverage containing the bifidobacterium breve CCFM1217 viable bacteria.

Other fermented foods including solid foods, liquid foods, semi-solid foods are prepared by fermentation production using bifidobacterium breve CCFM 1217. 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 able to reduce the levels of plasma TMAO, plasma TMA, cecal TMA; can improve the structure of intestinal flora, recover intestinal flora disorder caused by high choline, improve the abundance of beneficial bacteria (Roseburia, rikenella eae RC9 gun group), and reduce the relative abundance of harmful bacteria Anaeroplasma.

While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and 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 @ sBifidobacterium breve) CCFM1217 was deposited at the microorganism strain collection of Guangdong province at 12 months 31 of 2021 with the deposit number GDMCC No:62176.

2. a 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 10 ⁹ CFU/mL or ≡1X10. ⁹ CFU/g。

4. A pharmaceutical composition comprising bifidobacterium breve CCFM1217 of claim 1.

5. The pharmaceutical composition of claim 4, wherein the composition comprises a statin.

6. A fermented food product, characterized in that the fermented food product is a fermented food product comprising bifidobacterium breve CCFM 1217.

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

8. Use of bifidobacterium breve CCFM1217 as claimed in claim 1 in the manufacture of a medicament for reducing the risk of cardiovascular disease.

9. Use of bifidobacterium breve CCFM1217 as claimed in claim 1 in the manufacture of a medicament for reducing the risk of atherosclerosis.

10. The use according to claim 8 or 9, wherein the medicament further comprises a pharmaceutically acceptable carrier.