Bifidobacterium longum and application thereof
A technology of Bifidobacterium longum and Lactobacillus, applied in the direction of Bifidobacterium, application, Lactobacillus, etc., can solve the problems of unreachable treatment, side effects, allergies and liver diseases, and achieve non-toxic side effects, high expression level, expression The effect of level reduction
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Embodiment 1
[0029] Embodiment 1: the separation method of Bifidobacterium longum YS108R
[0030] The method for isolating Bifidobacterium longum, the main steps are as follows:
[0031] (1) The preparation of medium: added the MRS medium of 0.05% cysteine hydrochloride and 100mg / L mupirocin;
[0032] (2) After gradient dilution of the feces samples collected from healthy people, select appropriate 3 gradient dilutions, spread on the solid plate of the above medium, and place the plate upside down in an anaerobic incubator at 37°C for 48 hours;
[0033] (3) pick colonies with good wire-drawing effect and carry out streaking purification;
[0034] (4) Pick the purified colonies and inoculate them in liquid medium for anaerobic culture for 24-48 hours, collect the fermentation broth for the determination of exopolysaccharides, and keep a small amount of fermentation broth, collect the bacteria by centrifugation, and use them for genome extraction and PCR and 16S rDNA sequencing;
[0035...
Embodiment 2
[0040] Example 2: Determination of the properties of Bifidobacterium longum YS108R
[0041] Determination of biological characteristics of Bifidobacterium longum:
[0042] (1) Observation method of bacterial characteristics: pick a small amount of colony and smear it on a glass slide, carry out Gram staining, and observe under an optical microscope. Bacteria characteristics: Gram staining is positive, grows well in the environment of pH 3.0-8.0, does not form spores, the bacteria is about 0.5-1.5μm×2-8μm, rod-shaped, rod-shaped or bifurcated Y-shaped, transmission The results of electron microscope observation showed that there were obvious exopolysaccharides on the surface of the bacteria (attached figure 1 ).
[0043] (2) Observation method of colony characteristics: Streak the strain on the MRS solid medium, culture until a single colony grows, and observe the colony morphology.
[0044] Colony characteristics: Form round and raised colonies on MRS solid medium, milky wh...
Embodiment 3
[0051] Embodiment 3: the alleviating effect of Bifidobacterium longum YS108R on DSS-induced colitis mouse symptoms
[0052] Mouse pathology experiment, the main steps 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), 8 mice in each group, the experimental scheme and the treatment methods of mice in each group are shown in Table 2.
[0054] (2) After the above strains were cultured for 24 hours, the bacteria were collected by centrifugation, and the bacteria were resuspended with normal saline, and the number of viable bacteria was adjusted to 5×10 9 CFU / mL, made into a bacterial suspension, used for gavage of mice, and the gavage dose was 0.2mL. On the 14th day, dextran s...
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