Anti-aging bifidobacterium animalis and application thereof
An animal bifidobacteria, anti-aging technology, applied in the field of bioengineering, can solve problems such as few other diseases, and achieve the effect of increasing peroxidase activity, reducing brain inflammatory factors, and reversing anxiety-like behaviors
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Embodiment 1
[0030] Example 1: Acquisition and Identification of Animal Bifidobacteria
[0031] 1. Acquisition and identification of bifidobacteria
[0032] The feces of centenarians are stored in 50% glycerol, and diluted in sterile phosphate buffered saline (PBS). Select 3-5 appropriate gradients and draw 0.1ml into MRS medium for streak separation. Each dilution The gradient was done in two replicates. Put the petri dish into an incubator at 37°C for anaerobic culture for 48 hours, and then select smooth, convex, neat edges, and white colonies for microscopic examination. The suspected colonies were isolated and purified again in MRS medium according to the above steps until the colonies were completely purified. The purified Bifidobacterium animalis was continuously passaged for 3 times, and then stored at -80°C with a 1:1 mixture of 50% glycerol and bacterial liquid.
[0033] 2. Identification of Animal Bifidobacteria
[0034] After the selected bacterial colonies were continuousl...
Embodiment 2
[0036] Embodiment 2: the acid resistance experiment of animal bifidobacteria
[0037] Take out Bifidobacterium animalis from the ultra-low temperature refrigerator at -80°C and inoculate it in MRS medium, culture it anaerobically overnight at 37°C, then inoculate it into fresh MRS medium at 2%, culture it at 37°C for 24 hours, and dilute it with PBS buffer for 101 , 102, 103, 104, 105 times, centrifuge at 2500rpm for 4min, and discard the supernatant. Add PBS buffer solution with pH of 2, 3, 4, 5, 7, let stand for 4 hours, mix well, take 10uL to smear on the plate, culture in 37°C incubator anaerobically overnight, and count viable bacteria.
[0038] The result is as figure 1 As shown, Bifidobacterium animalis can tolerate PBS incubation with pH=3 and above, and has good acid resistance.
Embodiment 3
[0039] Embodiment 3: bile salt resistance test of animal bifidobacteria
[0040] MRS liquid medium with bile salt concentration of 0%, 0.1%, 0.2%, 0.3% and 0.5% bile salt was prepared in advance. Inoculate the frozen bifidobacteria in MRS medium, culture anaerobically at 37°C overnight, then inoculate them in MRS medium with different concentrations of bile salts at 2% and culture them for 24 hours, dilute them in PBS gradients, shake and mix well, Take 10uL to smear the plate, culture it anaerobically in a 37°C incubator overnight, and count the viable bacteria.
[0041] The result is as figure 2 As shown, Bifidobacterium animalis can survive in various concentrations of bile salt medium, showing good tolerance to bile salt.
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