A strain of Bifidobacterium breve capable of clearing toxins in the body and its postbiotics

Abstract

The present invention discloses a Bifidobacterium breve strain capable of clearing toxins in the body and its postbiotics, belonging to the technical field of microbial applications. The present invention isolates a Bifidobacterium breve strain ( Bifidobacterium breve ) WZA-BB07 from human feces, and its biological deposit number is: CCTCC NO: M20241530. The strain WZA-BB07 has good effects in degrading endotoxin and enterotoxin. The strain and Lactobacillus mucosae are mixed and fermented to prepare postbiotics. Bifidobacterium breve mainly improves the balance of intestinal flora and inhibits the growth of harmful bacteria. Lactobacillus mucosae helps repair the intestinal barrier through its metabolites during fermentation, enhancing the metabolism and clearance of toxins; extracts of Dendrobium officinale, Lonicera japonica, and Pueraria lobata are added to the fermentation medium, which not only provides rich nutrition for fermentation but also exerts the antioxidant, anti-inflammatory, and intestinal repair-promoting effects of ingredients with both medicinal and edible properties, further enhancing the detoxification ability and overall health effects of the postbiotic product.

Description

Technical Field

[0001] The present invention relates to the field of microbial technology, and particularly to a Bifidobacterium breve strain capable of removing toxins from the body and its postbiotics. Background Art

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

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

[0004] Meanwhile, the theory of homologous medicines and foods in traditional Chinese medicine emphasizes the natural regulation of toxin metabolism through daily diet. Extracts from homologous medicines and foods are derived from natural plants and have comprehensive effects of antioxidant, anti-inflammatory, repairing the intestinal barrier and regulating the flora, which can effectively assist in the metabolism and excretion of toxins. Combining the characteristics of postbiotics and extracts from homologous medicines and foods and exploring their synergistic effects can not only utilize the high-efficiency toxin degradation ability of postbiotics, but also play the systematic regulation role of extracts from homologous medicines and foods, forming a new solution for multi-pathway toxin management. Therefore, the combined application of extracts from homologous medicines and foods and postbiotics can not only improve the efficiency of toxin clearance, but also have higher safety and wide applicability. Summary of the Invention

[0005] Aiming at the above-mentioned existing technologies, the purpose of the present invention is to provide a Bifidobacterium breve strain capable of removing toxins from the body and its postbiotics.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions:

[0007] In the first aspect of the present invention, there is provided a Bifidobacterium breve strain capable of removing toxins from the body (Bifidobacterium breve ), WZA-BB07; This strain was deposited at the China Center for Type Culture Collection (abbreviated as CCTCC, address: within Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province) on July 10, 2024, and its biological deposit number is: CCTCC NO: M20241530.

[0008] In the second aspect of the present invention, a postbiotic of Bifidobacterium breve is provided, which is prepared by the following method:

[0009] (1) Inoculate the above-mentioned Bifidobacterium breve and Lactobacillus mucosae reuteri into a fermentation medium, and carry out fermentation culture to obtain a fermentation broth;

[0010] (2) Treat the fermentation broth at 100 - 120 °C for 20 - 40 min, centrifuge to obtain the supernatant, and dry it to obtain the postbiotic of Bifidobacterium breve.

[0011] Preferably, the deposit number of Lactobacillus mucosae reuteri is CCTCC NO: M20241390.

[0012] 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 mucosae reuteri is 3% - 7% of the volume of the fermentation medium.

[0013] Preferably, in step (1), the fermentation temperature is 38 - 41 °C, and the fermentation time is 48 - 72 h.

[0014] Preferably, in step (1), the fermentation medium uses the MRS liquid medium containing 0.5 g / L cysteine as the basal medium, and a medicinal and edible homologous extract with a volume fraction of 0.3% - 0.5% is added to the basal medium;

[0015] The medicinal and edible homologous extract is prepared by the following method:

[0016] Mix Dendrobium officinale, Lonicera japonica Thunb., and Pueraria lobata (Willd.) Ohwi evenly according to a mass ratio of 3:1:1, crush them to obtain a mixed material, add 20 - 30 times the mass of the mixed material of water, decoct for 1 - 3 h, cool down, and filter to obtain the medicinal and edible homologous extract.

[0017] In the third aspect of the present invention, the above-mentioned Bifidobacterium breve or postbiotic of Bifidobacterium breve is provided for use in the preparation of a drug for clearing toxins in the body.

[0018] Preferably, the toxin is endotoxin or enterotoxin.

[0019] The beneficial effects of the present invention:

[0020] The present invention screens a strain of Bifidobacterium breve with good effects on degrading endotoxin and enterotoxin, and co-ferments this strain with Lactobacillus mucosae to prepare postbiotics. The co-fermentation of Bifidobacterium breve and Lactobacillus mucosae gives full play to their respective advantages. Bifidobacterium breve mainly improves the balance of the intestinal flora and inhibits the growth of harmful bacteria, while Lactobacillus mucosae helps repair the intestinal barrier through its metabolites during fermentation, enhances the metabolism and clearance of toxins, and further improves the degradation efficiency. Extracts of Dendrobium officinale, Lonicera japonica, and Pueraria lobata are added to the fermentation medium, which not only provides rich nutrients for fermentation but also exerts the antioxidant, anti-inflammatory, and intestinal repair-promoting effects of the ingredients of both medicine and food, further enhancing the detoxification ability and overall health effects of the postbiotic product, providing an efficient, safe, and stable solution for the clearance of endotoxins in the body and the management of intestinal health, and having important application value and promotion prospects. Brief Description of the Drawings

[0021] Figure 1 : Weight changes of mice in different treatment groups during the period of toxin challenge;

[0022] Figure 2 : Measurement results of fecal water content of mice in different treatment groups at the end of toxin challenge. Detailed Description of the Embodiments

[0023] It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present application belongs.

[0024] As described in the background art, traditional toxin management methods mainly rely on drug treatment, but in the case of high toxin load or impaired intestinal function, the effects are limited, and long-term use of drugs may cause side effects. Based on this, the present invention screens a strain of Bifidobacterium breve that can clear endotoxins in the body, which has good degradation effects on endotoxin and enterotoxin. To further enhance its toxin degradation effect, the present invention co-ferments Bifidobacterium breve with Lactobacillus mucosae and combines it with extracts of ingredients of both medicine and food to prepare postbiotics, which has extremely high safety and stability and is particularly suitable for the development of functional drugs, providing an innovative and sustainable solution for toxin management.

[0025] In order to enable those skilled in the art to more clearly understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below with reference to specific embodiments.

[0026] The test materials used in the embodiments of the present invention are all conventional test materials in the art and can be obtained through commercial channels.

[0027] Among them, Lactobacillus mucosae ( Clostridium butyricumThe preservation number of () is CCTCC NO: M20241390. This strain belongs to the applicant and has been recorded in Patent CN118530915B.

[0028] The strain number of enterotoxigenic Escherichia coli ( Escherichia coli ETEC O78:K80 ) is CICC10413, which was purchased from China Center for Industrial Culture Collection.

[0029] The endotoxin (LPS, lipopolysaccharide) used in the present invention was purchased from Omer Biotechnology (Shanghai) Co., Ltd., and its titer is greater than or equal to 500000 EU / mg.

[0030] The staphylococcal enterotoxin B used in the present invention was purchased from Hubei Qifei Pharmaceutical Chemical Co., Ltd., with the product number QF3438, CAS number: 11100-45-1, and purity: 99%.

[0031] Example 1: Isolation and identification of strains:

[0032] The Bifidobacterium breve ( Bifidobacterium breve ) WZA-BB07 strain of the present invention was isolated from the feces of healthy adults.

[0033] 1. Isolation of strains:

[0034] Taking the feces of healthy adults as samples, 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 physiological saline (containing 0.5 g / L cysteine) under sterile conditions to obtain a 10 -1 dilution. The above dilution steps were repeated to successively obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 dilutions; 100 μL of different gradient dilutions were respectively pipetted and spread on MRS solid medium (containing 0.5 g / L cysteine), and cultured at 37 °C for 72 h to obtain dilution-coated plates; the typical colonies on the dilution-coated plates were picked and streaked on MRS solid medium (containing 0.5 g / L cysteine) respectively, and cultured at 37 °C for 48 h to obtain purified colonies; the 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.

[0035] 2. Strain identification:

[0036] By sending the WZA-BB07 strain to the Henan Engineering Technology Research Center for Industrial Microbial Strains for 16S rDNA gene sequence identification, and comparing the sequencing results by BLAST-N in NCBI, it shows thatBifidobacterium breve The species sequence identity reached 100%, and the results showed that this strain belongs to Bifidobacterium breve ( Bifidobacterium breve ). And this strain was deposited, and the deposit information is as follows:

[0037] Species name: Bifidobacterium breve WZA-BB07

[0038] Latin name: Bifidobacterium breve

[0039] Deposit institution: China Center for Type Culture Collection

[0040] Abbreviation of deposit institution: CCTCC

[0041] Address: Inside Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province

[0042] Deposit date: July 10, 2024

[0043] Deposit number: CCTCC NO: M20241530.

[0044] Example 2: Performance test of Bifidobacterium breve WZA-BB07:

[0045] Preparation of the fermentation broth of Bifidobacterium breve WZA-BB07:

[0046] Inoculate Bifidobacterium breve WZA-BB07 into MRS solid medium (containing 0.5 g / L cysteine) for activation, and inoculate the activated Bifidobacterium breve into MRS liquid medium (containing 0.5 g / L cysteine) at an inoculation amount of 10% (volume fraction), and culture at 37 °C for 48 h. Subsequently, centrifuge the culture solution (4,000 rpm, 10 minutes), discard the supernatant, collect the precipitated cells, resuspend the cells with sterile physiological saline (0.85% NaCl), and adjust the concentration of the cell suspension to 1×10 9 cfu / mL.

[0047] 1. Performance test of Bifidobacterium breve WZA-BB07 for degrading endotoxin:

[0048] Treatment group: Inoculate the cell suspension of Bifidobacterium breve WZA-BB07 into MRS liquid medium (containing 0.5 g / L cysteine) at an inoculation amount of 10% (volume fraction), add 1 mL of lipopolysaccharide with 100 EU to the MRS liquid medium (containing 0.5 g / L cysteine), and react with shaking at 35 °C for 2 h;

[0049] Blank control group: Select a blank MRS liquid medium (containing 0.5 g / L cysteine), add 1 mL of 100 EU lipopolysaccharide to the MRS liquid medium (containing 0.5 g / L cysteine), and perform an oscillating reaction at 35 °C for 2 h;

[0050] Use the endotoxin detection horseshoe crab kit from Shanghai Enzyme-linked Biotechnology Co., Ltd. to test the concentration of lipopolysaccharide after the reaction in the treatment group and the blank control group. Each test is set with three replicates.

[0051] Endotoxin degradation rate (%) = (endotoxin content in the blank control group - endotoxin content in the treatment group) / endotoxin content in the blank control group × 100.

[0052] 2. Performance test of Bifidobacterium breve WZA - BB07 in degrading enterotoxin:

[0053] Treatment group: Inoculate the bacterial suspension of Bifidobacterium breve WZA - BB07 into the MRS liquid medium (containing 0.5 g / L cysteine) at an inoculation amount of 10% (volume fraction), add 1 mL of 100 TU staphylococcal enterotoxin B to the MRS liquid medium (containing 0.5 g / L cysteine), and perform an oscillating reaction at 35 °C for 2 h;

[0054] The blank control group selects the MRS liquid medium (containing 0.5 g / L cysteine), adds 1 mL of 100 TU staphylococcal enterotoxin B to the MRS liquid medium (containing 0.5 g / L cysteine), and performs an oscillating reaction at 35 °C for 2 h;

[0055] Use an ELISA detection kit to test the concentration of staphylococcal enterotoxin B after the reaction in the treatment group and the blank control group. Each test is set with three replicates.

[0056] Staphylococcal enterotoxin B degradation rate (%) = (staphylococcal enterotoxin B content in the blank control group - staphylococcal enterotoxin B content in the treatment group) / staphylococcal enterotoxin B content in the blank control group × 100.

[0057] 3. Test results:

[0058] Table 1 Determination of the effect of Bifidobacterium breve WZA - BB07 on detoxifying toxins

[0059]

[0060] As can be seen from Table 1, Bifidobacterium breve WZA - BB07 has a good degradation effect on endotoxin and staphylococcal enterotoxin B.

[0061] Example 3: Preparation of Bifidobacterium breve postbiotics:

[0062] (1) Preparation of fermentation medium:

[0063] 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;

[0064] The medicinal and edible extract is prepared by the following method:

[0065] 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;

[0066] (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 8 cfu / 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;

[0067] (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.

[0068] Example 4: Preparation of Bifidobacterium breve postbiotics:

[0069] (1) Preparation of fermentation medium:

[0070] 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;

[0071] The medicinal and edible extract is prepared by the following method:

[0072] 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;

[0073] (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;

[0074] (3) Treat the fermentation broth at 110 °C for 30 min, centrifuge to obtain the supernatant, and dry it to obtain postbiotics of Bifidobacterium breve.

[0075] Example 5: Preparation of postbiotics of Bifidobacterium breve:

[0076] (1) Preparation of the fermentation medium:

[0077] Add the extract of medicine and food homology with a volume fraction of 0.5% to the MRS liquid medium (containing 0.5 g / L cysteine) to prepare the fermentation medium;

[0078] The extract of medicine and food homology is prepared by the following method:

[0079] Mix Dendrobium officinale, honeysuckle, and kudzu root evenly according to the mass ratio of 3:1:1, crush them to obtain a mixture, add 25 times the mass of water of the mixture, decoct for 2 h, cool down, and filter to obtain the extract of medicine and food homology;

[0080] (2) Inoculate the activated Bifidobacterium breve seed liquid (the viable count is 1.5×10 8 cfu / mL) at an inoculation amount of 13% (volume fraction), and the activated Limosilactobacillus reuteri seed liquid (the viable count is 1.5×10 8 cfu / mL) at an inoculation amount of 7% (volume fraction) into the fermentation medium, and carry out fermentation culture at 35 °C for 48 h to obtain the fermentation broth;

[0081] (3) Treat the fermentation broth at 110 °C for 30 min, centrifuge to obtain the supernatant, and dry it to obtain postbiotics of Bifidobacterium breve.

[0082] Comparative Example 1: Preparation of postbiotics of Bifidobacterium breve:

[0083] (1) Preparation of the fermentation medium:

[0084] Add the extract of medicine and food homology with a volume fraction of 0.5% to the MRS liquid medium (containing 0.5 g / L cysteine) to prepare the fermentation medium;

[0085] The extract of medicine and food homology is prepared by the following method:

[0086] Mix Dendrobium officinale, honeysuckle, and kudzu root evenly according to the mass ratio of 3:1:1, crush them to obtain a mixture, add 25 times the mass of water of the mixture, decoct for 2 h, cool down, and filter to obtain the extract of medicine and food homology;

[0087] (2) The activated Bifidobacterium breve seed liquid (the viable count is 1.5×10 8Inoculate the fermentation medium at an inoculation amount of 20% (volume fraction) with a viable count of

[0088] (3)Treat the fermentation broth at 110 °C for 30 min, centrifuge to obtain the supernatant, and dry it to obtain postbiotics.

[0089] Comparative Example 2:

[0090] (1)Preparation of the fermentation medium:

[0091] Add 0.5% (volume fraction) of the medicine and food homology extract to the MRS liquid medium (containing 0.5 g / L cysteine) to prepare the fermentation medium;

[0092] The medicine and food homology extract is prepared by the following method:

[0093] Mix Dendrobium officinale, honeysuckle, and kudzu root evenly according to the mass ratio of 3:1:1, crush them to obtain a mixture, add 25 times the mass of water of the mixture, decoct for 2 h, cool down, and filter to obtain the medicine and food homology extract;

[0094] (2)Inoculate the activated Limosilactobacillus reuteri seed liquid (the viable count is 1.5×10 8 cfu / mL) into the fermentation medium at an inoculation amount of 20% (volume fraction), and carry out fermentation culture at 35 °C for 48 h to obtain the fermentation broth;

[0095] (3)Treat the fermentation broth at 110 °C for 30 min, centrifuge to obtain the supernatant, and dry it to obtain postbiotics.

[0096] Comparative Example 3:

[0097] (1)Preparation of the fermentation medium:

[0098] Add 0.5% (volume fraction) of the medicine and food homology extract to the MRS liquid medium (containing 0.5 g / L cysteine) to prepare the fermentation medium;

[0099] The medicine and food homology extract is prepared by the following method:

[0100] Crush Dendrobium officinale and add 25 times the mass of water, decoct for 2 h, cool down, and filter to obtain the medicine and food homology extract;

[0101] (2)The activated Brevibacterium breve seed liquid (the viable count is 1.5×10 8 cfu / mL) at an inoculation amount of 13% (volume fraction), and the activated Limosilactobacillus reuteri seed liquid (the viable count is 1.5×10 8Inoculate into the fermentation medium at an inoculum size of 7% (volume fraction) at a concentration of cfu / mL, and perform fermentation culture at 35°C for 48 h to obtain a fermentation broth;

[0102] (3)Treat the fermentation broth at 110°C for 30 min, centrifuge to obtain the supernatant, and dry to obtain postbiotics.

[0103] Control Example 4:

[0104] (1)Preparation of the fermentation medium:

[0105] Add 0.5% (volume fraction) of the medicine and food homology extract to the MRS liquid medium (containing 0.5 g / L cysteine) to prepare the fermentation medium;

[0106] The medicine and food homology extract is prepared by the following method:

[0107] Crush honeysuckle and add 25 times its mass of water, decoct for 2 h, cool down, and filter to obtain the medicine and food homology extract;

[0108] (2)The activated Bifidobacterium breve seed liquid (viable cell count is 1.5×10 8 cfu / mL) is inoculated into the fermentation medium at an inoculum size of 13% (volume fraction), and the activated Limosilactobacillus reuteri seed liquid (viable cell count is 1.5×10 8 cfu / mL) is inoculated into the fermentation medium at an inoculum size of 7% (volume fraction), and perform fermentation culture at 35°C for 48 h to obtain a fermentation broth;

[0109] (3)Treat the fermentation broth at 110°C for 30 min, centrifuge to obtain the supernatant, and dry to obtain postbiotics.

[0110] Control Example 5:

[0111] (1)Preparation of the fermentation medium:

[0112] Add 0.5% (volume fraction) of the medicine and food homology extract to the MRS liquid medium (containing 0.5 g / L cysteine) to prepare the fermentation medium;

[0113] The medicine and food homology extract is prepared by the following method:

[0114] Crush kudzu root and add 25 times its mass of water, decoct for 2 h, cool down, and filter to obtain the medicine and food homology extract;

[0115] (2)The activated Bifidobacterium breve seed liquid (viable cell count is 1.5×10 8 cfu / mL) is inoculated into the fermentation medium at an inoculum size of 13% (volume fraction), and the activated Limosilactobacillus reuteri seed liquid (viable cell count is 1.5×10 8Inoculate (cfu / mL) into the fermentation medium at an inoculum size of 7% (v / v), and perform fermentation culture at 35 °C for 48 h to obtain the fermentation broth;

[0116] (3) Treat the fermentation broth at 110 °C for 30 min, centrifuge to collect the supernatant, and dry to obtain postbiotics.

[0117] Test Example 1:

[0118] Conduct experiments according to the method in 2.3 of the reference document "Study on the alleviating effect and mechanism of different Lactobacilli on diarrhea caused by enterotoxigenic Escherichia coli [D]. Jiangnan University, 2021.".

[0119] 1. Preparation of enterotoxigenic Escherichia coli suspension:

[0120] Inoculate the activated enterotoxigenic Escherichia coli into nutrient broth medium (NB), and culture it on a shaker at 37 °C for 12 - 18 hours until the culture broth becomes turbid, indicating that the bacteria have grown sufficiently. Subsequently, centrifuge the culture broth (4,000 rpm, 10 minutes), discard the supernatant, and collect the precipitated bacteria. Resuspend the bacteria with sterile physiological saline (0.85% NaCl), and adjust the concentration of the bacterial suspension to 1.2×10 11 cfu / mL.

[0121] 2. Grouping of experimental animals:

[0122] Take BALB / c mice. After the mice are purchased, adaptively raise them in the animal house for 5 days. During this period, they can drink water and eat freely, and all mice are fed with basal diet. After the adaptive feeding of the mice is completed, randomly divide them into 7 groups, with 9 mice in each group.

[0123] At the beginning of the experiment, add streptomycin (5 g / L) to the drinking water of all mice for 72 h to eliminate the normal intestinal flora. During this period, all mice are fed with basal diet. Then fast for 18 h, and perform challenge after 18 h. Each treatment group is intragastrically administered with enterotoxigenic Escherichia coli suspension, and the intragastric administration dose is 0.1 mL / kg of mouse body weight, twice a day for 4 days. Starting from the first day of challenge, each treatment group is fed in the following ways:

[0124] Model control group: Only feed basal diet;

[0125] Treatment 1: Feed basal diet and the postbiotics of Bifidobacterium breve prepared in Example 5, and add 1 g of Bifidobacterium breve postbiotics to every 1 kg of basal diet;

[0126] Treatment 2: Feed basal diet and the postbiotics prepared in Comparative Example 1, and add 1 g of postbiotics to every 1 kg of basal diet;

[0127] Treatment 3: Feed the basal diet and the postbiotics prepared in Comparative Example 2, adding 1 g of postbiotics per 1 kg of basal diet;

[0128] Treatment 4: Feed the basal diet and the postbiotics prepared in Comparative Example 3, adding 1 g of postbiotics per 1 kg of basal diet;

[0129] Treatment 5: Feed the basal diet and the postbiotics prepared in Comparative Example 4, adding 1 g of postbiotics per 1 kg of basal diet;

[0130] Treatment 6: Feed the basal diet and the postbiotics prepared in Comparative Example 5, adding 1 g of postbiotics per 1 kg of basal diet.

[0131] Feed continuously according to the above test method for 14 days, and then euthanize all mice.

[0132] Composition of the basal diet: 70% corn flour, 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.

[0133] 3. Test indicators:

[0134] 3.1 Mouse body weight change:

[0135] Record the body weight regularly every day, and statistically analyze the body weight changes of mice in each group during the challenge period, and take the average value.

[0136] 3.2 Determination of mouse fecal water content:

[0137] Collect the feces of each mouse at the end of the challenge, then perform freeze-drying to measure the water content in the feces, and take the average value.

[0138] Fecal water content (%) = (fecal wet weight - fecal dry weight) / (fecal wet weight) × 100.

[0139] 3.3 Detection of mouse heat-stable enterotoxin (ST):

[0140] After euthanizing the mice, open the abdominal cavity in a sterile environment, expose the intestine, take out the jejunum part, gently squeeze the intestinal contents into a sterile centrifuge tube with your finger, centrifuge at 3000 - 5000 rpm for 10 minutes at 4°C to obtain the serum and aliquot it, and then store it at -80°C. According to the manufacturer's instructions, use the ELISA kit from Shanghai Enzyme-linked Biotechnology to measure the content of heat-stable enterotoxin (ST) in the serum, and take the average value.

[0141] Table 2 Detection of heat-stable enterotoxin (ST) in mice of different treatment groups

[0142]

[0143] It can be seen from Figure 1 and Figure 2 that feeding the postbiotics of Bifidobacterium breve prepared in Example 5 of the present invention to Treatment 1 can alleviate the weight loss and the increase in fecal water content caused by infection with enterotoxigenic Escherichia coli in mice; Treatments 2-3 feed the postbiotics prepared by fermenting single Bifidobacterium breve and Lactobacillus mucosae roiae respectively, which can also significantly alleviate the weight loss and the increase in fecal water content caused by infection with enterotoxigenic Escherichia coli 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 mucosae roiae to prepare postbiotics has a synergistic effect on alleviating the weight loss and the increase in fecal water content caused by infection with enterotoxigenic Escherichia coli in mice. Treatments 4-6 feed the postbiotics prepared by adding a single medicine and food homology extract to MRS liquid medium (containing 0.5 g / L cysteine) respectively. The body weights of the mice are all lower than those of the postbiotics of Bifidobacterium breve prepared in Example 5, and the fecal water contents of the mice are all higher than those of the postbiotics of Bifidobacterium breve prepared in Example 5, indicating that the compounding of Dendrobium officinale, Lonicera japonica and Pueraria lobata in the present invention to prepare a medicine and food homology extract and adding it to the MRS medium for fermentation has a synergistic effect on alleviating the weight loss caused by infection with enterotoxigenic Escherichia coli in mice.

[0144] It can be seen from Table 2 that there are obvious differences in the content of heat-stable enterotoxin (ST) among the mice in different treatment groups. By comparing the ST contents (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 clearing ST, and its ST content is the lowest, only 0.03 pg / mL, indicating that the postbiotics have the strongest degradation ability for heat-stable enterotoxin (ST) and can effectively clear the heat-stable enterotoxin (ST) in the body and reduce the inflammatory reaction caused by heat-stable enterotoxin (ST). Treatments 2-3 feed the postbiotics prepared by fermenting single Bifidobacterium breve and Lactobacillus mucosae roiae respectively, and the heat-stable enterotoxin (ST) contents are 238.7 pg / mL and 512.8 pg / mL respectively. Although they are significantly lower than those of the model control group, they are still higher than those of Treatment 1, indicating that the postbiotics of single strains are not as effective as the combined ones in degrading ST. The heat-stable enterotoxin (ST) contents of Treatments 4-6 (adding a single medicine and food homology extract to the medium) are 487.2 pg / mL to 543.2 pg / mL, indicating that the postbiotics fermented only with a single medicine and food homology extract are not very effective in degrading heat-stable enterotoxin (ST), indicating that the compounding of Dendrobium officinale, Lonicera japonica and Pueraria lobata in the present invention to prepare a medicine and food homology extract and adding it to the MRS medium for fermentation has a synergistic effect on clearing the heat-stable enterotoxin (ST) in the body of mice.

[0145] The above are only the preferred embodiments of the present application and are not intended to limit the present application. For those skilled in the art, various changes and modifications can be made to the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included within 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 drug for removing toxins from an organism, characterized in that: The toxin is lipopolysaccharide, staphylococcal enterotoxin B or heat-stable enterotoxin.

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