Microbial agent and application of microbial agent to prevention and treatment of constipation
A technology of microbial inoculum and microbial strains is applied in the field of microorganisms to achieve the effects of good application compliance, inhibiting intestinal peristalsis and reducing symptoms of constipation
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Embodiment 1
[0021] Embodiment 1 Preparation of microbial agent of the present invention (Bacillus amyloliquefaciens bacterium liquid or powder)
[0022] 1. Take out the strain preservation tube, scratch the plate with LB solid medium for recovery, and incubate at 37°C for 48 hours. Pick a single colony under the plate and inoculate it into 50 ml of LB medium, and culture it in an incubator with shaking at 37°C for 24 hours. The seeds are inoculated with 5% inoculation amount into a 5L large triangle flask with a liquid volume of 2LLB medium, shaken and cultured at 37°C for 20-28 hours, and the concentration of the bacteria solution is tested. The viable bacteria content exceeds 2 billion / ml, and it is ready As a liquid strain of Bacillus amyloliquefaciens;
[0023] 2. Use the liquid Bacillus amyloliquefaciens cultured in step 1 as seeds, inoculate into 600L fermentation medium, the inoculation amount is 10%, stir at 120r / min, the air volume per minute liquid-gas ratio is 1:1, and cultiva...
Embodiment 2
[0026] Example 2 The safety test of the microbial agent of the present invention (Bacillus amyloliquefaciens)
[0027]The Bacillus amyloliquefaciens liquid that the embodiment of the present invention 1 produces is through the rat acute toxicity test in Shaoyang College, and the conclusion result shows that the Bacillus amyloliquefaciens liquid is harmful to rats (feeding the Bacillus amyloliquefaciens liquid 14 of the present invention by 0.6ml / 10g body weight) day) Effect: All rats survived, moved freely, had smooth hair, normal diet, no secretions in the respiratory tract, eyes and mouth, no symptoms of poisoning were found, and their body weight increased. Compared with the control group of the blank group, the body weight of the rats fed with the Bacillus amyloliquefaciens solution increased significantly (P<0.001). At the end of the 14-day observation period, the rats were dislocated and dissected, and no obvious abnormalities were found in the heart, liver, spleen, lung...
Embodiment 3
[0028] Example 3 Bacillus amyloliquefaciens prevents and treats constipation in mice
[0029] Establishment of mouse constipation model
[0030] After adapting to the feeding environment for one week, the mice were subcutaneously injected with 5 mg / kg loperamide hydrochloride (Loperamide hydrochloride) physiological saline suspension, once a day at 9:00 and 18:00, for 7 consecutive days, and 20 of them were selected as The normal group was given the same dose of normal saline. Select mice with dry feces, reduced number, and reduced weight as constipated mice.
[0031] Grouping and Dosage
[0032] Select 80 mice with successful modeling and randomly divide them into model groups (Constipation, whole intestine Bacillus licheniformis group (after separating and purifying the whole intestine strain produced by Shenyang First Pharmaceutical Co., Ltd. of Northeast Pharmaceutical Group Company, centrifuge after pure-bred fermentation, The fermentation medium is the same as the...
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