A kind of bifidobacterium longum and its application
A technology of Bifidobacterium longum and Lactobacillus, applied in the direction of Bifidobacterium, Applied, Lactobacillus, etc., can solve the problems of side effects, allergies, liver diseases, and failure to achieve treatment, and achieve high expression level, lower expression level, no Toxic effects
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Embodiment 1
[0029] Example 1: Isolation method of Bifidobacterium longum YS108R
[0030] The main steps of the isolation method of Bifidobacterium longum are as follows:
[0031] (1) Preparation of culture medium: MRS culture medium supplemented with 0.05% cysteine hydrochloride and 100 mg / L mupirocin;
[0032] (2) After gradient dilution of the collected stool samples of healthy people, select appropriate 3 gradient dilutions and spread them on the solid plate of the above-mentioned medium, and place the plate upside down in an anaerobic incubator at 37°C for 48 hours;
[0033] (3) Pick the colonies with good wire drawing effect for streaking purification;
[0034] (4) Pick the purified colonies and inoculate them in liquid culture medium for anaerobic culture for 24 to 48 hours. Collect the fermentation broth for the determination of extracellular polysaccharides, while retaining a small amount of fermentation broth. After centrifugation to collect the bacteria, it is used for genome extraction...
Embodiment 2
[0040] Example 2: Determination of the properties of Bifidobacterium longum YS108R
[0041] Determination of the biological characteristics of Bifidobacterium longum:
[0042] (1) Observation method of bacterial characteristics: pick a small amount of colonies and apply them on a glass slide, perform Gram staining, and observe under an optical microscope. Bacterial characteristics: Gram staining is positive, it grows well under pH 3.0-8.0 environment, does not form spores, the bacterial cell is about 0.5-1.5μm×2-8μm, it is rod-shaped, rod-shaped or bifurcated Y-shaped, transmission The electron microscope observation results show that there are obvious extracellular polysaccharides on the surface of the bacteria (attached figure 1 ).
[0043] (2) Observation method of colony characteristics: streak the strain on the MRS solid medium, cultivate until a single bacteria grows, and observe its colony morphology.
[0044] Colony characteristics: round, convex colonies formed on MRS solid ...
Embodiment 3
[0051] Example 3: Relief effect of Bifidobacterium longum YS108R on the symptoms of DSS-induced colitis in mice
[0052] The main steps of mouse pathology experiment are as follows:
[0053] (1) Prepare 40 male C57 mice and randomly divide them into 5 groups: normal control group (Control), model group (Model), Bifidobacterium longum YS108R intervention group (YS108R+DSS), Bifidobacterium longum C11A10B intervention group (C11A10B+DSS) and the commercial strain Bifidobacterium lactis BB12 intervention group (BB12+DSS), each with 8 mice. The experimental protocol and the treatment of mice in each group are shown in Table 2.
[0054] (2) After culturing the above-mentioned strains for 24 hours respectively, collect the cells by centrifugation, resuspend the cells in physiological saline, and adjust the number of viable cells to 5×10 9 CFU / mL, made into bacterial suspension, used for gavage mice, the gavage dose is 0.2mL. On the 14th day, dextran sodium sulfate (DSS) was made into a 2....
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