Bifidobacterium breve capable of removing toxins in body and metagen of bifidobacterium breve

Abstract

The invention discloses a strain of Bifidobacterium breve capable of removing toxins in a body and a metagen of the Bifidobacterium breve, and belongs to the technical field of microbial application, the Bifidobacterium breve WZA-BB07 is separated from human excrement, and the biological preservation number of the Bifidobacterium breve WZA-BB07 is CCTCC (China Center for Type Culture Collection) NO: M20241530. The bacterial strain WZA-BB07 has a good effect of degrading endotoxin and enterotoxin, the bacterial strain and lactobacillus reuteri are subjected to mixed fermentation to prepare the postbiotics, bifidobacterium breve mainly improves intestinal flora balance and inhibits growth of harmful bacteria, the lactobacillus reuteri helps repair intestinal barriers through metabolites of the lactobacillus reuteri in the fermentation process, and the lactobacillus reuteri and lactobacillus reuteri are mixed to prepare the postbiotics. Metabolism and removal of toxins are enhanced; dendrobium officinale, honeysuckle and radix puerariae extracts are added into a fermentation culture medium, so that rich nutrition is provided for fermentation, the effects of resisting oxidation, resisting inflammation and promoting intestinal repair of medicinal and edible components are also exerted, and the detoxification capacity and the overall health effect of the postbiotic product are further improved.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a strain of Bifidobacterium breve capable of removing toxins from an organism and a postbiotic thereof. Background Art

[0002] In recent years, due to environmental pollution, unreasonable diet structure and changes in lifestyle, the problem of toxin accumulation in the human body has become increasingly prominent. For example, the increased levels of endotoxin (LPS) and enterotoxin are closely related to a variety of chronic diseases (such as metabolic syndrome, non-alcoholic fatty liver disease, and inflammatory bowel disease). Traditional toxin management methods mainly rely on drug therapy, but the effect is limited in the case of high toxin load or impaired intestinal function, and long-term use of drugs may cause side effects.

[0003] As a natural means of toxin regulation, probiotics play an important role in improving the balance of intestinal flora, enhancing intestinal barrier and immune function. However, the stability of live probiotics in transportation, storage and gastrointestinal environment is poor, which is greatly affected by individual differences and the effect is unstable. At the same time, some people (such as those with low immunity) have certain restrictions on the application of live bacteria. Postbiotics, as an extension of the application of probiotics, overcome the stability and safety issues of live probiotics with its metabolites and inactivated bacteria as the core. Postbiotics can achieve efficient management of toxins by directly binding to toxins, repairing intestinal barriers and regulating immune responses, which is especially suitable for people with damaged intestinal barriers or high toxin loads. However, it may be difficult to fully cover the toxin management needs by relying solely on postbiotics.

[0004] At the same time, the theory of food and medicine in traditional Chinese medicine emphasizes the natural regulation of toxin metabolism through daily diet. Food and medicine extracts are derived from natural plants and have the comprehensive effects of anti-oxidation, anti-inflammatory, repairing intestinal barriers and regulating flora, which can effectively assist the metabolism and excretion of toxins. Combining the characteristics of postbiotics and food and medicine extracts, exploring the synergistic effect of the two can not only utilize the efficient toxin degradation ability of postbiotics, but also give play to the systemic regulatory effect of food and medicine extracts, forming a new solution for multi-pathway toxin management. Therefore, the combination of food and medicine extracts and postbiotics can not only improve the efficiency of toxin removal, but also have higher safety and wide applicability. Summary of the invention

[0005] In view of the above prior art, the purpose of the present invention is to provide a strain of Bifidobacterium breve and its postbiotics that can remove toxins in the body.

[0006] To achieve the above object, the present invention adopts the following technical solutions: In a first aspect of the present invention, a strain of Bifidobacterium breve ( Bifidobacterium breve)WZA-BB07; the strain was deposited in the China Center for Type Culture Collection (CCTCC for short, address: Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province) on July 10, 2024, and its biological preservation number is: CCTCC NO: M20241530.

[0007] A second aspect of the present invention provides a Bifidobacterium breve postbiotic, which is prepared by the following method: (1) inoculating the above-mentioned Bifidobacterium breve and Lactobacillus reuteri into a fermentation medium for fermentation to obtain a fermentation liquid; (2) The fermentation broth is treated at 100-120° C. for 20-40 min, the supernatant is centrifuged and dried to obtain Bifidobacterium breve postbiotics.

[0008] Preferably, the deposit number of Lactobacillus reuteri is CCTCC NO: M20241390.

[0009] Preferably, in step (1), the inoculation amount of Bifidobacterium breve is 7%-13% of the volume of the fermentation medium, and the inoculation amount of Lactobacillus reuteri is 3%-7% of the volume of the fermentation medium.

[0010] Preferably, in step (1), the fermentation temperature is 38-41°C and the fermentation time is 48-72h.

[0011] Preferably, in step (1), the fermentation medium uses MRS liquid medium containing 0.5 g / L cysteine ​​as the basic medium, and 0.3%-0.5% by volume of the medicinal and edible extract is added to the basic medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are evenly mixed in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 20-30 times the mass of the mixture, boiled for 1-3 hours, cooled, filtered, and obtained a medicinal and edible extract.

[0012] The third aspect of the present invention provides the use of the above-mentioned Bifidobacterium breve or Bifidobacterium breve postbiotics in the preparation of a product for removing toxins from an organism.

[0013] Preferably, the toxin is an endotoxin or an enterotoxin.

[0014] Preferably, the product is food or medicine.

[0015] Beneficial effects of the present invention: The present invention screens out a strain of Bifidobacterium breve with good effects of degrading endotoxins and enterotoxins, and mixes the strain with Lactobacillus reuteri for fermentation to prepare postbiotics. The synergistic fermentation of Bifidobacterium breve and Lactobacillus reuteri gives full play to their respective advantages. Bifidobacterium breve mainly improves the balance of intestinal flora and inhibits the growth of harmful bacteria. Lactobacillus reuteri helps repair the intestinal barrier through its metabolites during the fermentation process, enhances the metabolism and removal of toxins, and further improves the degradation efficiency. Adding extracts of Dendrobium officinale, Honeysuckle and Pueraria lobata to the fermentation medium not only provides rich nutrition for fermentation, but also exerts the antioxidant, anti-inflammatory and intestinal repair promoting effects of the medicinal and edible homologous ingredients, further enhances the detoxification ability and overall health effect of the postbiotic product, and provides an efficient, safe and stable solution for the removal of toxins in the body and the management of intestinal health, which has important application value and promotion prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 : Body weight changes of mice in different treatment groups during the challenge period; Figure 2 : Results of fecal water content determination of mice in different treatment groups at the end of virus challenge. DETAILED DESCRIPTION

[0017] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0018] As described in the background technology, traditional toxin management methods mainly rely on drug therapy, but in the case of high toxin load or impaired intestinal function, the effect is limited, and long-term use of drugs may cause side effects. Based on this, the present invention screened a strain of Bifidobacterium breve that can remove toxins from the body, which has a good degradation effect on endotoxins and enterotoxins. In order to further enhance its toxin degradation effect, the present invention prepares postbiotics by co-fermenting Bifidobacterium breve with Lactobacillus reuteri and combining them with medicinal and edible extracts. The postbiotics have extremely high safety and stability, are particularly suitable for the development of functional foods and drugs, and provide an innovative and sustainable solution for toxin management.

[0019] In order to enable those skilled in the art to more clearly understand the technical solution of the present application, the technical solution of the present application will be described in detail below in conjunction with specific embodiments.

[0020] The test materials used in the examples of the present invention are all conventional test materials in the art and can be purchased through commercial channels.

[0021] Among them, Lactobacillus reuteri ( Clostridium butyricum) is deposited with CCTCC NO: M20241390, which is owned by the applicant and has been recorded in patent CN118530915B.

[0022] Enterotoxigenic Escherichia coli ( Escherichia coli ETEC O78:K80 ) strain number is CICC10413 and was purchased from China Industrial Microbiological Culture Collection Center.

[0023] The endotoxin (LPS, lipopolysaccharide) used in the present invention was purchased from Omer Biotechnology (Shanghai) Co., Ltd., and the potency was greater than or equal to 500,000 EU / mg.

[0024] The staphylococcal enterotoxin B used in the present invention was purchased from Hubei Qifei Pharmaceutical Chemical Co., Ltd. with a product number of QF3438, a CAS number of 11100-45-1, and a purity of 99%.

[0025] Example 1: Isolation and identification of strains: The present invention Bifidobacterium breve Bifidobacterium breve ( Bifidobacterium breve ) The WZA-BB07 strain was isolated from the feces of a healthy adult.

[0026] 1. Isolation of strains: Using healthy adult feces as a sample, 0.5 g of the sample stored in 30% (v / v) glycerol was added to a 10 mL centrifuge tube containing 4.5 mL of normal saline (containing 0.5 g / L cysteine) under sterile conditions to obtain 10 -1 Dilution, repeat the above dilution steps to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 dilution; 100 μL of gradient dilutions of different gradients were respectively spread on MRS solid medium (containing 0.5 g / L cysteine), and cultured at 37°C for 72 h to obtain dilution spread plates; typical colonies on the dilution spread plates were picked and streaked on MRS solid medium (containing 0.5 g / L cysteine), and cultured at 37°C for 48 h to obtain purified colonies; purified colonies were picked and inoculated into MRS liquid medium (containing 0.5 g / L cysteine), and cultured at 37°C for 48 h to obtain strain WZA-BB07.

[0027] 2. Strain identification: The WZA-BB07 strain was sent to the Henan Industrial Microbial Engineering Technology Research Center for 16SrDNA gene sequence identification. The sequencing results were compared with the BLAST-N of NCBI. Bifidobacterium breveThe species sequence identity reached 100%, and the results showed that the strain belonged to Bifidobacterium breve ( Bifidobacterium breve ). The strain was biologically deposited, and the deposit information is as follows: Bacterial species name: Bifidobacterium breve WZA-BB07 Latin name: Bifidobacterium breve Collection institution: China Center for Type Culture Collection Abbreviation of depository institution: CCTCC Address: Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province Date of deposit: July 10, 2024 Deposit number: CCTCC NO: M20241530.

[0028] Example 2: Performance test of Bifidobacterium breve WZA-BB07: Preparation of Bifidobacterium breve WZA-BB07 fermentation broth: Bifidobacterium breve WZA-BB07 was inoculated into MRS solid medium (containing 0.5 g / L cysteine) for activation. The activated Bifidobacterium breve was inoculated into MRS liquid medium (containing 0.5 g / L cysteine) at an inoculum rate of 10% (volume fraction) and cultured at 37°C for 48 h. Subsequently, the culture solution was centrifuged (4,000 rpm, 10 min), the supernatant was discarded, the precipitated bacteria were collected, and the bacteria were resuspended in sterile saline (0.85% NaCl) to adjust the concentration of the bacterial suspension to 1×10 9 cfu / mL.

[0029] 1. Performance test of Bifidobacterium breve WZA-BB07 in degrading endotoxin: Treatment group: The bacterial suspension of Bifidobacterium breve WZA-BB07 was inoculated into MRS liquid medium (containing 0.5 g / L cysteine) at a volume fraction of 10%, and 1 mL of 100 EU lipopolysaccharide was added to the MRS liquid medium (containing 0.5 g / L cysteine), and the reaction was shaken at 35°C for 2 h. Blank control group: select blank MRS liquid culture medium (containing 0.5 g / L cysteine), add 1 mL 100 EU lipopolysaccharide to MRS liquid culture medium (containing 0.5 g / L cysteine), and shake at 35°C for 2 h; The endotoxin detection horseshoe crab kit of Shanghai ELISA Biological Company was used to test the concentration of lipopolysaccharide in the treatment group and the blank control group after reaction, and three replicates were set for each test. Endotoxin degradation rate (%) = (endotoxin content in blank control group - endotoxin content in treatment group) / endotoxin content in blank control group × 100.

[0030] 2. Performance test of Bifidobacterium breve WZA-BB07 in degrading enterotoxins: Treatment group: The bacterial suspension of Bifidobacterium breve WZA-BB07 was inoculated into MRS liquid medium (containing 0.5 g / L cysteine) at a volume fraction of 10%, and 1 mL 100 TU of Staphylococcus enterotoxin B was added to the MRS liquid medium (containing 0.5 g / L cysteine), and the reaction was shaken at 35°C for 2 h. The blank control group used MRS liquid medium (containing 0.5 g / L cysteine), and 1 mL 100 TU of Staphylococcus enterotoxin B was added to the MRS liquid medium (containing 0.5 g / L cysteine), and the reaction was shaken at 35°C for 2 h; The concentration of Staphylococcus enterotoxin B after the reaction in the treatment group and the blank control group was tested using an ELISA test kit, and three replicates were set for each test. Staphylococcal enterotoxin B degradation rate (%) = (Staphylococcal enterotoxin B content in blank control group - Staphylococcal enterotoxin B content in treatment group) / Staphylococcal enterotoxin B content in blank control group × 100.

[0031] 3. Test results: Table 1 Determination of the toxin degradation effect of Bifidobacterium breve WZA-BB07 As shown in Table 1, Bifidobacterium breve WZA-BB07 has a good degradation effect on endotoxin and Staphylococcal enterotoxin B.

[0032] Example 3: Preparation of Bifidobacterium breve postbiotics: (1) Preparation of fermentation medium: Adding 0.3% by volume of the edible-drug extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are mixed evenly in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 25 times the mass of the mixture, boiled for 2 hours, cooled, filtered, and obtained a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable bacteria count 3×10 8 cfu / mL) at a 7% (volume fraction) inoculation volume, activated Lactobacillus reuteri seed solution (viable count 3×10 8cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 3% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth was treated at 110°C for 30 min, the supernatant was collected by centrifugation, and dried to obtain Bifidobacterium breve postbiotics.

[0033] Example 4: Preparation of Bifidobacterium breve postbiotics: (1) Preparation of fermentation medium: Adding 0.4% by volume of the edible-drug extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are mixed evenly in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 25 times the mass of the mixture, boiled for 2 hours, cooled, filtered, and obtained a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable count 2×10 8 cfu / mL) at a 10% (volume fraction) inoculation volume, activated Lactobacillus reuteri seed solution (viable count 2×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 5% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth was treated at 110°C for 30 min, the supernatant was collected by centrifugation, and dried to obtain Bifidobacterium breve postbiotics.

[0034] Example 5: Preparation of Bifidobacterium breve postbiotics: (1) Preparation of fermentation medium: Adding 0.5% by volume of the edible-medicinal extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are mixed evenly in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 25 times the mass of the mixture, boiled for 2 hours, cooled, filtered, and obtained a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable cell count 1.5×10 8 cfu / mL) according to the inoculation volume of 13% (volume fraction), the activated Lactobacillus reuteri seed solution (the number of viable bacteria was 1.5×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 7% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth was treated at 110°C for 30 min, the supernatant was collected by centrifugation, and dried to obtain Bifidobacterium breve postbiotics.

[0035] Comparative Example 1: Preparation of Bifidobacterium breve postbiotics: (1) Preparation of fermentation medium: Adding 0.5% by volume of the edible-medicinal extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are mixed evenly in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 25 times the mass of the mixture, boiled for 2 hours, cooled, filtered, and obtained a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable cell count 1.5×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 20% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth is treated at 110°C for 30 min, the supernatant is collected by centrifugation, and dried to obtain postbiotics.

[0036] Comparative Example 2: (1) Preparation of fermentation medium: Adding 0.5% by volume of the edible-medicinal extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are mixed evenly in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 25 times the mass of the mixture, boiled for 2 hours, cooled, filtered, and obtained a medicinal and edible extract; (2) The activated Lactobacillus reuteri seed solution (viable count 1.5×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 20% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth is treated at 110°C for 30 min, the supernatant is collected by centrifugation, and dried to obtain postbiotics.

[0037] Comparative Example 3: (1) Preparation of fermentation medium: Adding 0.5% by volume of the edible-medicinal extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Crush the Dendrobium officinale, add 25 times the mass of water, boil for 2 hours, cool down, filter, and obtain a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable cell count 1.5×10 8 cfu / mL) according to the inoculation volume of 13% (volume fraction), the activated Lactobacillus reuteri seed solution (the number of viable bacteria was 1.5×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 7% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth is treated at 110°C for 30 min, the supernatant is collected by centrifugation, and dried to obtain postbiotics.

[0038] Comparative Example 4: (1) Preparation of fermentation medium: Adding 0.5% by volume of the edible-medicinal extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Crush honeysuckle and add 25 times the weight of water, boil for 2 hours, cool down, filter, and obtain a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable cell count 1.5×10 8 cfu / mL) according to the inoculation volume of 13% (volume fraction), the activated Lactobacillus reuteri seed solution (the number of viable bacteria was 1.5×10 8 cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 7% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth is treated at 110°C for 30 min, the supernatant is collected by centrifugation, and dried to obtain postbiotics.

[0039] Comparative Example 5: (1) Preparation of fermentation medium: Adding 0.5% by volume of the edible-medicinal extract to an MRS liquid medium (containing 0.5 g / L cysteine) to prepare a fermentation medium; The medicinal and edible extract is prepared by the following method: Grind the kudzu root, add 25 times the mass of water, boil for 2 hours, cool down, filter, and obtain a medicinal and edible extract; (2) The activated Bifidobacterium breve seed solution (viable cell count 1.5×10 8 cfu / mL) according to the inoculation volume of 13% (volume fraction), the activated Lactobacillus reuteri seed solution (the number of viable bacteria was 1.5×10 8cfu / mL) was inoculated into the fermentation medium at an inoculum rate of 7% (volume fraction), and the fermentation culture was carried out at 35°C for 48 hours to obtain a fermentation broth; (3) The fermentation broth is treated at 110°C for 30 min, the supernatant is collected by centrifugation, and dried to obtain postbiotics.

[0040] Test Example 1: The experiment was conducted according to the method 2.3 in the literature "Study on the alleviating effect and mechanism of different lactobacilli on diarrhea caused by enterotoxigenic Escherichia coli [D]. Jiangnan University, 2021."

[0041] 1. Preparation of enterotoxigenic Escherichia coli suspension: The activated enterotoxigenic E. coli was inoculated into nutrient broth (NB) and cultured in a shaking incubator at 37°C for 12-18 hours until the culture medium became turbid, indicating that the bacteria had fully grown. Subsequently, the culture medium was centrifuged (4,000 rpm, 10 minutes), the supernatant was discarded, and the precipitated bacteria were collected. The bacteria were resuspended in sterile saline (0.85% NaCl) and the concentration of the bacterial suspension was adjusted to 1.2×10 11 cfu / mL.

[0042] 2. Experimental animal grouping: BALB / c mice were purchased and adaptively raised in the animal room for 5 days, during which they were free to drink water and eat. All mice were fed with basic feed. After the adaptive feeding of mice, they were randomly divided into 7 groups, with 9 mice in each group.

[0043] At the beginning of the experiment, streptomycin (5g / L) was added to the drinking water of all mice for 72 hours to eliminate the normal intestinal flora. During this period, all mice were fed with basic feed. Then they were fasted for 18 hours. After 18 hours, they were challenged with toxins. Each treatment group was gavaged with enterotoxigenic Escherichia coli suspension at a gavage volume of 0.1mL / kg mouse body weight, twice a day, for 4 days. Starting from the first day of challenge, each treatment group was fed according to the following method: Model control group: only fed with the basic diet; Treatment 1: feeding the basic feed and the Bifidobacterium breve postbiotics prepared in Example 5, with 1 g of Bifidobacterium breve postbiotics added per 1 kg of the basic diet; Treatment 2: feeding the basal feed and the postbiotics prepared in Comparative Example 1, with 1 g of postbiotics added to every 1 kg of basal diet; Treatment 3: feeding the basal feed and the postbiotics prepared in Comparative Example 2, with 1 g of postbiotics added to every 1 kg of basal diet; Treatment 4: feeding the basal feed and the postbiotics prepared in Comparative Example 3, with 1 g of postbiotics added to every 1 kg of basal diet; Treatment 5: feeding the basal feed and the postbiotics prepared in Comparative Example 4, with 1 g of postbiotics added to every 1 kg of basal diet; Treatment 6: The rats were fed with the basic feed and the postbiotics prepared in Comparative Example 5, with 1 g of postbiotics added to every 1 kg of the basic diet.

[0044] The mice were fed continuously for 14 days according to the above experimental method, and then all mice were euthanized. Basic feed ingredients: 70% corn meal, 1.8% soybean meal, 17% fish meal, 5% grass meal, 2% yeast powder, 1.9% vegetable oil, 0.4% essential amino acids, 0.8% vitamins, 0.7% minerals, 0.2% CaHPO4, 0.2% Nacl.

[0045] 3. Test indicators: 3.1 Changes in mouse weight: The body weight was recorded regularly every day, and the weight changes of each group of mice during the infection period were counted and the average value was taken.

[0046] 3.2 Determination of water content in mouse feces: At the end of the challenge, the feces of each mouse were collected and freeze-dried to measure the water content in the feces and take the average value.

[0047] Feces moisture content (%) = (feces wet weight - feces dry weight) / (feces wet weight) × 100.

[0048] 3.3 Detection of heat-resistant enterotoxin (ST) in mice: After the mice were euthanized, the abdominal cavity was opened under sterile conditions to expose the intestines, the jejunum was removed, and the intestinal contents were gently squeezed into a sterile centrifuge tube with fingers, centrifuged at 3000-5000 rpm for 10 minutes at 4°C, serum was obtained and aliquoted, and then stored at -80°C. The content of heat-stable enterotoxin (ST) in serum was measured using the ELISA kit of Shanghai ELISA Biotechnology according to the manufacturer's instructions, and the average value was taken.

[0049] Table 2 Detection of heat-resistant enterotoxin (ST) in mice in different treatment groups Depend on Figure 1 and Figure 2It can be seen that feeding the postbiotics of Bifidobacterium breve prepared in Example 5 of the present invention to treatment 1 can alleviate the weight loss and increased fecal water content caused by enterotoxigenic Escherichia coli infection in mice; treatments 2-3 were respectively fed with postbiotics prepared by fermentation of single Bifidobacterium breve and Lactobacillus reuteri, which can also significantly alleviate the weight loss and increased fecal water content caused by enterotoxigenic Escherichia coli infection in mice, but there is still a certain gap compared with the mice in treatment 1, indicating that the mixed fermentation of Bifidobacterium breve and Lactobacillus reuteri to prepare postbiotics has a synergistic effect on alleviating the weight loss and increased fecal water content caused by enterotoxigenic Escherichia coli infection in mice. Treatments 4-6 were respectively fed with postbiotics prepared by adding a single edible and medicinal extract to MRS liquid culture medium (containing 0.5 g / L cysteine). The weights of the mice were lower than those of the mice fed with the Bifidobacterium breve postbiotics prepared in Example 5, and the water content of the feces of the mice was higher than that of the mice fed with the Bifidobacterium breve postbiotics prepared in Example 5, indicating that the present invention has a synergistic effect in alleviating the weight loss caused by enterotoxigenic Escherichia coli infection in mice by compounding the edible and medicinal extracts of Dendrobium officinale, Honeysuckle and Pueraria lobata.

[0050] As shown in Table 2, there are significant differences in the content of heat-stable enterotoxin (ST) in mice in different treatment groups. By comparing the ST content (pg / mL) of each treatment group, it can be seen that treatment 1 (feeding the postbiotics of Bifidobacterium breve prepared in Example 5 of the present invention) has a significant advantage in removing ST, and its ST content is the lowest, only 0.03 pg / mL, indicating that the postbiotic has the strongest ability to degrade heat-stable enterotoxin (ST), and can effectively remove heat-stable enterotoxin (ST) in the body and reduce the inflammatory response caused by heat-stable enterotoxin (ST). Treatments 2-3 were fed with postbiotics prepared by fermentation of single Bifidobacterium breve and Lactobacillus reuteri, and the content of heat-stable enterotoxin (ST) was 238.7 pg / mL and 512.8 pg / mL, respectively, which was significantly lower than the model control group, but still higher than treatment 1, indicating that the postbiotics of a single strain are not as effective as the combination in degrading ST. The content of heat-stable enterotoxin (ST) in treatments 4-6 (adding a single edible and medicinal extract to the culture medium) was 487.2 pg / mL to 543.2 pg / mL, indicating that the effect of postbiotics fermented with only a single edible and medicinal extract on the degradation of heat-stable enterotoxin (ST) was not ideal, indicating that the present invention prepared a edible and medicinal extract by compounding Dendrobium officinale, Honeysuckle and Pueraria lobata and adding it to MRS culture medium for fermentation had a synergistic effect in clearing heat-stable enterotoxin (ST) in mice.

[0051] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A strain of Bifidobacterium breve that can remove toxins from the body ( Bifidobacterium breve )WZA-BB07, characterized by, Its biological deposit number is: CCTCC NO: M20241530.

2. A Bifidobacterium breve postbiotic, characterized in that Prepared by the following method: (1) inoculating the Bifidobacterium breve and Lactobacillus reuteri described in claim 1 into a fermentation medium for fermentation to obtain a fermentation liquid; (2) The fermentation broth is treated at 100-120° C. for 20-40 min, the supernatant is centrifuged and dried to obtain Bifidobacterium breve postbiotics.

3. The Bifidobacterium breve postbiotic according to claim 2, characterized in that The deposit number of Lactobacillus reuteri is CCTCC NO: M20241390.

4. The Bifidobacterium breve postbiotic according to claim 2, characterized in that In step (1), the inoculation amount of Bifidobacterium breve is 7%-13% of the volume of the fermentation medium, and the inoculation amount of Lactobacillus reuteri is 3%-7% of the volume of the fermentation medium.

5. The Bifidobacterium breve postbiotic according to claim 2, characterized in that In step (1), the fermentation temperature is 38-41°C and the fermentation time is 48-72h.

6. The Bifidobacterium breve postbiotic according to claim 2, characterized in that In step (1), the fermentation medium uses MRS liquid medium containing 0.5 g / L cysteine ​​as the basic medium, and 0.3%-0.5% by volume of the edible and medicinal extract is added to the basic medium; The medicinal and edible extract is prepared by the following method: Dendrobium officinale, honeysuckle and kudzu root are evenly mixed in a mass ratio of 3:1:1, crushed to obtain a mixture, added with water 20-30 times the mass of the mixture, boiled for 1-3 hours, cooled, filtered, and obtained a medicinal and edible extract.

7. Use of the Bifidobacterium breve according to claim 1 or the Bifidobacterium breve postbiotic according to claim 2 in the preparation of a product for removing toxins from an organism.

8. The use according to claim 7, characterized in that: The toxin is an endotoxin or an enterotoxin.

9. The use according to claim 7, characterized in that: The product is a food or a medicine.

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