Forage clostridium butyricum, microecological preparation with function of promoting livestock and poultry growth and preparation method of microecological preparation
A micro-ecological preparation, the technology of Clostridium butyricum, is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., which can solve the problem of tea fermentation probiotic strains lacking tea polyphenols, unable to achieve application effects, and excessive tea. Phenol oxidation loss and other problems, to achieve the effect of enhancing immunity, improving body immunity, and maintaining digestive function
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Embodiment 1
[0073] Embodiment 1--Isolation and identification of Clostridium butyricum RCM-2
[0074] 1. Isolation, enrichment and purification of Clostridium butyricum RCM-2
[0075] Under aseptic operation conditions, farmers in Xia Village, Zherong County, Ningde City, Fujian Province (coordinates: 119 。 52′04.1″E, 27 。 13′32.7″N) The cecal mucosal segment of a healthy adult rooster bred in a closed mountain forest, the mucosal mucus was scraped with a sterilized glass slide and placed in an EP tube, diluted with sterile normal saline for 10 2 Then heated to 60°C for 30min, took 0.1mL and spread it on the RCM agar plate containing bromocresol purple indicator (0.01%, g / mL), after coating, put the RCM agar plate in the anaerobic incubator , cultured at 37°C for 48 hours, selected the colonies that turned yellow in the medium, and purified them by streaking, and selected the rod-shaped strains that were positive for Gram staining and had spores in the microscopic examination, and conta...
Embodiment 2
[0086] Embodiment 2--clostridium butyricum RCM-2 anaerobic and aerobic fermentation in MRS liquid medium
[0087](1) Activation of Clostridium butyricum RCM-2: Wash the slant of the Clostridium butyricum RCM-2 test tube with sterile saline in a sterile environment, wash off the bacterial lawn on the surface of the medium, and then inoculate it into the MRS liquid In the culture medium, culture at 37°C for 48 hours;
[0088] (2) Anaerobic static fermentation of Clostridium butyricum RCM-2 in MRS liquid medium:
[0089] Inoculate the activation solution of Clostridium butyricum RCM-2 obtained in step (1) into an anaerobic serum bottle containing MRS liquid medium in an anaerobic box according to the inoculation amount of 3% (volume ratio), and incubate statically at 37°C for 48 hours. At the end of the culture, the number of live bacteria, spore rate and pH value of the fermentation broth were measured;
[0090] (3) Aerobic fermentation of Clostridium butyricum RCM-2 in MRS li...
Embodiment 3
[0094] Embodiment 3--The effect of Pseudostellaria ginseng extract on the growth and pH of Clostridium butyricum RCM-2
[0095] (1) Activation of Clostridium butyricum RCM-2: Same as step (1) in Example 2;
[0096] (2) Inoculate the activation solution of Clostridium butyricum RCM-2 obtained in step (1) into an anaerobic serum bottle containing MRS liquid medium in an anaerobic box according to the inoculation amount of 3% (volume ratio). The gradient of Radix Pseudostellariae Pseudostellaria extract in the MRS liquid medium in the serum bottle was successively set to (g / mL): 0%, 0.10%, 0.25%, 0.5%, cultured at 37°C for 48 hours, and the fermentation broth was measured at the end of the culture pH value and number of viable bacteria, the results are shown in Table 3;
[0097]
[0098] (3) Put the activation solution of Clostridium butyricum RCM-2 obtained in step (1) into a 500 mL Erlenmeyer flask filled with 100 mL of MRS liquid medium according to the inoculation amount ...
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