Bifidobacterium longum subsp. longum for relieving constipation and increasing total bile acid content of excrement and application of Bifidobacterium longum subsp. longum
A technology of Bifidobacterium longum and subspecies, applied in the field of microorganisms, can solve problems such as single function, difficult activation, and slow growth, and achieve good growth characteristics, avoid side effects, and obvious effects
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[0073] (4) Preparation of Bifidobacterium longum suspension
[0074] Inoculate the 3-generation activated bacterial solution into 1L of liquid MRS medium at an inoculum size of 2%, shake and mix well, and incubate in an anaerobic incubator at 37°C for 24h. Centrifuge at 8000g / min for 15min at 4°C, remove the supernatant, wash twice with sterile saline containing 0.05%-0.1% L-cysteine hydrochloride, and centrifuge under the same conditions. After the supernatant was removed, it was stored in 30% glycerol and stored in a -80°C refrigerator for one week. Before the animal experiment, centrifuge the bacterial solution at 6000r / min for 5min, wash it twice with sterile saline, resuspend the bacterial solution with 10% skimmed milk, shake it evenly, and use the plate pouring method to measure the initial and one week after freezing the number of live bacteria. The formula of MRS medium is: beef extract 10g; tryptone 10g; yeast powder 5g; glucose 20g; anhydrous sodium acetate 5g; ...
Embodiment 1
[0076] Example 1: Bifidobacterium longum CCFM1113 relieves constipation-related symptoms induced by loperamide
[0077] Bifidobacterium longum CCFM1113, Bifidobacterium bifidum CGMCC NO.13632 (the strain is disclosed in the patent document CN106834187B) and Bifidobacterium longum CCFM642 (the strain is disclosed in the paper DOI: 10.3389 / fmicb.2019.01721 Middle) strains were taken out from the -80°C refrigerator, streaked on the MRS plate, cultured at 37°C for 48 hours, picked a single colony in the MRS liquid tube, cultured at 37°C for 24 hours, and inoculated on a new MRS at a volume of 2%. In liquid culture medium, cultivate at 37°C for 24 hours, culture for another generation in the same way, then centrifuge the bifidobacterium suspension at 6000r / min, 4°C for 5min, and then resuspend with 10% skimmed milk, Bacterial suspensions were obtained for animal experiments.
[0078] Thirty 6-week-old healthy male Balb / c mice were taken, adapted to the environment for 1 week, and ...
Embodiment 2
[0087] Embodiment 2: Bifidobacterium longum CCFM1113 can significantly improve the content of total bile acid (TBA) in feces of mice with constipation
[0088] Balb / c mice grouping, modeling and treatment methods were the same as in Example 1. The feces collected and freeze-dried on the 37th day were pre-treated: take 20 mg of dried mouse feces, add 1 mL of tert-butanol solution (volume ratio 1:100), vortex and mix, and then vortex after 15 minutes in a 37°C water bath Mix well for 10 seconds, and finally centrifuge at 4°C, 10,000×g for 2 minutes to take the supernatant. The total TBA content in the supernatant was determined by using the total TBA kit according to the relevant content of the instructions.
[0089] Total bile acids (TBA) are a group of metabolites of cholesterol in the liver and entero-hepatic circulation, and are mainly divided into two categories: primary bile acids and secondary bile acids. Changes in total bile acid content indicate to a certain extent t...
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