A strain of Bifidobacterium longum and its application
A technology of bifidobacteria longum and bifidobacteria is applied in the field of microorganisms to achieve the effects of strong adhesion, improved intestinal propulsion rate, and good growth and acid production capacity.
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Embodiment 1
[0038]Example 1: Screening of Bifidobacterium longum CCFM 760
[0039] 1) Use a disposable sterile toilet to collect several parts of fresh feces from a two-year-old girl in Binhu District, Wuxi City, Jiangsu Province, put the feces samples in a modified MRS liquid test tube with fructooligosaccharides as the carbon source, and place them in an anaerobic workstation ( N 2 :CO 2 :H 2 =80:10:10) to carry out the enrichment of bifidobacterium strains within 12h;
[0040] 2) Perform gradient dilution with sterile normal saline, use a pipette to draw 100 μL of the corresponding dilution sample and spread it on the modified MRS solid plate containing 0.02g / mL calcium carbonate, and incubate anaerobically for 24-48h;
[0041] 3) Pick a single colony with calcium carbonate calcium dissolution circle, and single colony that conforms to the basic form of lactic acid bacteria is streaked and separated on the improved MRS solid medium, and the streak is repeated 3 times;
[0042] 4) P...
Embodiment 2
[0046] Embodiment 2: the preparation of Bifidobacterium longum CCFM760 bacterial suspension
[0047] Experimental method: Inoculate the 3-generation activated bacterial solution into 1L of the modified MRS medium at an inoculum size of 2%, shake and mix well, and culture anaerobically at 37°C for 20 hours. Centrifuge at 5000 rpm and 4°C for 15 minutes, discard the supernatant, wash twice with sterile saline, centrifuge under the same conditions, and resuspend with 15 mL of sterile 30% sucrose water. After oscillating evenly, measure the number of viable bacteria with the plate pouring method, and store in a -80°C refrigerator for 1 week. Among them, the formula of the improved MRS medium is: beef extract 10g; tryptone 10g; yeast powder 5g; glucose 20g; anhydrous sodium acetate 5g; 4 ·7H 2 O 0.1g; MnSO 4 ·H 2 O 0.05g; Diammonium hydrogen citrate 2g; K 2 HPO 4 ·3H 2 O 2.6g; Tween-801mL; L-cysteine hydrochloride 1g; adjust the pH to 6.8±0.2; dilute to 1L. Sterilize at 1...
Embodiment 3
[0049] Example 3: Relief effect of Bifidobacterium longum CCFM760 on constipation in mice
[0050] Experimental method: The CCFM760 bacterial suspension with the function of relieving constipation in Example 1 was used as the experimental group, and Bifidobacterium animalis BB12 and phenolphthalein were used as the positive controls, and a comparative experiment was carried out on normal mice and constipation mice respectively. The effect of Bifidobacterium longum CCFM760 on alleviating constipation in mice was evaluated by comparing the water content of feces, the time of first black stool discharge, the propulsion rate of small intestine, the content of constipation-related gastrointestinal peptide in colon tissue sections and serum of mice in each group.
[0051] Take 25 healthy BALB / c male mice (7 weeks old) weighing 22-26g, and divide them into 5 groups randomly (normal control group, constipation model group, CCFM760 experimental group, BB12 control group, phenolphthalein...
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