Complex probiotic and application thereof
A technology of compound probiotics and bifidobacteria, which is applied in the field of microorganisms, can solve the problems of few researches on the compound formula of probiotics
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[0043] The present invention also provides a preparation method for the composite probiotics, in which Bifidobacterium adolescentis CCFM8630, Lactobacillus reuteri CCFM8631, Lactobacillus rhamnosus CCFM1044 and Lactobacillus casei CCFM711 are respectively inoculated into the improved MRS medium, Cultivate under anaerobic conditions at 34-39°C for 18-24 hours, collect the bacteria; resuspend with freeze-drying protectant to make the content of each bacteria reach 10 10 CFU / mL or more, then the suspension was cultured under anaerobic conditions at 37°C for 40-60 minutes, dried to obtain freeze-dried bacterial powder of each bacterial cell, and the number of viable bacteria in the bacterial powder was confirmed by resuspension and dilution plate coating; The Bifidobacterium adolescentis CCFM8630 freeze-dried bacteria powder, Lactobacillus reuteri CCFM8631 freeze-dried bacteria powder, Lactobacillus rhamnosus CCFM1044 and Lactobacillus casei CCFM711 freeze-dried bacteria powder wer...
Embodiment 1
[0076] Example 1: Compound probiotics have good tolerance to simulated gastrointestinal fluid
[0077] Bifidobacterium adolescentis CCFM8630, Lactobacillus reuteri CCFM8631, Lactobacillus rhamnosus CCFM1044 and Lactobacillus casei CCFM711 stored at -80°C were inoculated in the modified MRS medium (mMRS) respectively. Cultivate at 37°C for 36 hours, then subculture twice with an inoculation amount of 2%-4%, adjust the concentration of viable bacteria in the culture solution of Bifidobacterium adolescentis CCFM8630, Lactobacillus reuteri CCFM8631, Lactobacillus rhamnosus CCFM1044 and Lactobacillus casei CCFM711 5×10 8 CFU / mL, take the concentration as 5×10 8 CFU / mL of Bifidobacterium adolescentis CCFM8630, Lactobacillus reuteri CCFM8631, Lactobacillus rhamnosus CCFM1044 and Lactobacillus casei CCFM711 culture solution 1mL each, mix well, take 1mL of mixed culture solution and 9.0mL pH 2.5 artificial gastric juice (mMRS medium containing 1% pepsin, pH=2.5) mixed, cultured in an...
Embodiment 2
[0084] Example 2: Compound probiotics have no toxic side effects on C57BL / 6J mice
[0085] Suspend the compound probiotic bacteria with the ratio of Bifidobacterium adolescentis CCFM8630, Lactobacillus reuteri CCFM8631, Lactobacillus rhamnosus CCFM1044 and Lactobacillus casei CCFM711 with a ratio of 1:1:1:1 viable bacteria in 3% sucrose solution , made at a concentration of 1.0×10 9 CFU / mL bacterial suspension. Eight healthy male C57BL / 6J mice with a body weight of about 20g-22g were taken, and after one week of adaptive feeding, they were fed with the bacterial suspension of this concentration once a day, observed for one week, and the death and body weight were recorded. The experimental results are shown in Table 3.
[0086] Table 3 Changes in body weight and death of mice
[0087] time (days)
1
2
3
4
5
6
7
weight(g)
21.1±0.9
21.3±1.1
21.5±1.3
21.6±1.2
21.9±0.8
22.2±0.8
22.5±0.9
Circumstances of death
-...
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