A common strain of Bacteroides vulgaris that relieves acute intestinal injury caused by lipopolysaccharide
A technology of common Bacteroides and bacterial agents, applied in the field of microorganisms, to achieve the effect of wide application prospects
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Embodiment 1
[0053] Example 1: Isolation and screening of Bacteroides vulgaris CCFM1152
[0054] 1. Sample collection
[0055] A fecal sample was collected from a 50-year-old middle-aged man in Dieqiao Apartment, Hedong District, Tianjin. The sample was placed in a stool tube filled with 30% glycerol, and kept in a thermal insulation box with an ice pack. After returning to the laboratory, it was quickly placed in- 80 ℃ refrigerator to be separated and screened.
[0056] 2. Isolation and purification of strains
[0057] (1) Dilution coating: take about 0.5 g of the content stored in 30% glycerol and add it to a 10 mL centrifuge tube containing 4.5 mL of normal saline in a sterile environment to obtain 10 -1 Diluent, repeat the above dilution steps to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 Diluent;
[0058] (2) Coating culture: Pipette 100 μL of 10 in step (1). -4 , 10 -5 , 10 -6 Three gradient dilutions were placed on the Bacteroides-specific screening medium, spread evenl...
Embodiment 2
[0073] Example 2: Regulation of regulatory T cell content in mesenteric lymph nodes of lipopolysaccharide-infected mice by Bacteroides vulgaris CCFM1152
[0074] Specific steps are as follows:
[0075] 1. Preparation of Bacteroides vulgaris CCFM1152 intragastric fluid
[0076] The Bacteroides vulgaris CCFM1152 obtained in step 2 of Example 1 was streaked in BHI solid medium, and cultivated under anaerobic conditions at 37°C for 48 hours to obtain a single colony; the single colony on the BHI solid medium was picked and inoculated into BHI liquid medium medium, enrichment culture under anaerobic conditions at 37°C for 18-20 hours for activation, and continuous activation for two generations to obtain an activated culture; the activated culture is inoculated into BHI liquid culture at an inoculum amount of 2% to 5% (v / v). In the base, enriched and cultured under anaerobic conditions at 37°C for 18 to 20 hours to obtain bacterial liquid, centrifuged at 8000 g for 10 min to obtai...
Embodiment 3
[0083]Example 3: Effect of Bacteroides vulgaris CCFM1152 on intestinal pathological state of inflammatory mice
[0084] Specific steps are as follows:
[0085] The specific implementation method is the same as that in Example 2. Mice were sacrificed and part of their colon tissues were taken and placed in 4% paraformaldehyde solution for observation of tissue sections. The hematoxylin-eosin staining method was used to evaluate the lipopolysaccharide stimulation-induced colon tissue damage in mice and the protective effect of Bacteroides vulgaris intervention on colon tissue.
[0086] The experimental results are as figure 2 The results showed that there were inflammatory infiltration areas and goblet cell apoptosis in the colon tissue sections of the mice in the modeling group; after the intervention of Bacteroides vulgaris CCFM1152, the intestinal intestinal status of the inflammatory mice was significantly improved, and there was almost no inflammatory infiltration and muc...
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