Lactobacillus fermentum zhao capable of regulating intestinal movement and preventing constipation
A technology of fermenting Lactobacillus and intestinal motility, applied in the field of microorganisms, to achieve the effect of increasing food intake and drinking water, reducing defecation volume, and speeding up the time of black stool discharge
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Embodiment 1
[0055] Example 1: Isolation, purification and preliminary identification of Lactobacillus fermentum zhao.
[0056] This embodiment is carried out according to the following steps:
[0057] (1) Separation and purification of Lactobacillus fermentum zhao.
[0058] Aseptically draw 500 μL of the sample and add it to 5 mL of sterilized physiological saline to make a uniform dilution of 1:10, and continue to make a certain proportion of dilution. Select an appropriate gradient dilution solution, pipette 100 μL each into the MRS solid plate medium with a sterile pipette tip, and incubate at 30°C for 48-72 hours, observe and record the colony morphology, such as figure 1 shown. Use an inoculation loop (or a sterilized toothpick) to pick different colonies from the surface and interior of the plate and inoculate them in the MRS liquid medium, and culture them on a shaker at 30°C and 300r / min for 24-48h; repeat the above steps for continuous activation for 2 generations before coatin...
Embodiment 2
[0063] Example 2: In vitro screening of Lactobacillus fermentum zhao.
[0064] (1) Screening of probiotics resistant to pH 3.0 artificial gastric juice.
[0065] Preparation of artificial gastric juice: 0.2% NaCl, 0.35% pepsin, adjust the pH value to 3.0 with 1M HCl, filter and sterilize with a vacuum pump in an aseptic operating table for later use.
[0066] Determination of the tolerance of probiotics to artificial gastric juice: take 5mL of the activated strain culture solution, pour it into a sterilized 10mL centrifuge tube in a sterile operating table, collect the bacteria by centrifugation at 3000r / min for 10min, add 5mL of sterilized Mix physiological saline to make bacterial suspension, take 1mL bacterial suspension and 9mL artificial gastric juice with pH3. Samples were taken, poured into MRS agar medium and cultured at 37°C for 48h. The number of viable bacteria was determined by plate counting method, and the survival rate (%) was calculated: survival rate (%)=num...
Embodiment 3
[0080] Example 3: Activated carbon induced constipation experiment.
[0081] Feed healthy Kunming mice, 6 weeks old, weighing 23±2g, 50 females. After feeding the basal diet for 5 days, the mice were randomly divided into 5 groups according to body weight, namely: normal group, constipation control group, bisacodyl group, Lactobacillus bulgaricus group, and high and low intragastric doses of Lactobacillus fermentum zhao group, 10 in each group. Raise them in stainless steel cages, keep room temperature at 24±2°C, relative humidity at 50±10%, and light-dark rotation for 12 hours (lighting at 8:00-20:00). During the whole experimental period, the normal group and the constipation control group were fed with the basal diet, and the mice in the experimental group were fed with 1.0×10 9 CFU / kg gavage probiotics. After 2 weeks, at 9:00 every morning, except the normal group, the other 5 groups of mice were gavaged with 10% activated carbon ice water at 2°C, 0.2 mL / only, once a da...
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