Microbial inoculant and application thereof in livestock breeding
A microbial agent, a technology for livestock and poultry breeding, applied in the field of microorganisms, can solve the problems of high price, insufficient effect, and inability to completely replace antibiotics, and achieve the effects of high safety, reduced breeding costs, and shortened breeding time
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Embodiment 1
[0017] The isolation of embodiment 1 bacillus licheniformis
[0018] In December 2014, the Bacillus licheniformis strain was isolated from the intestinal contents of wild wild boars in Tantou Town, Longhui County, Hunan Province. The specific method is: take the intestinal contents of wild wild boar and add it to a conical flask filled with 10 mL of sterilized water, shake and incubate for 30 min (200 r / min), then take 1 mL of the suspension and place it in 9 mL of sterile water to make 100 mL of sterile water. -2 Dilute the suspension, and so on, to make 10 -5 , 10 -6 Dilute the suspension; draw 100 μL each, add it to the nutrient broth medium plate, MRS medium, and PDA medium, and spread it evenly with a coating bar. Each treatment is repeated 3 times. Place the above culture dish in a constant temperature incubator at 37°C for 1-2 days (PDA medium is cultured at 30°C). Select a single colony and transfer it to the corresponding medium for culture. After the colony grows,...
Embodiment 2
[0020] Example 2 Bacillus licheniformis to the in vitro antibacterial effect of common pathogenic bacteria
[0021] The specific experimental method is as follows:
[0022] 1, YHSH-02 Preparation of bacterial suspension: get slant seeds, inoculate into 50ml sterilized TSB liquid culture medium, shake and cultivate at constant temperature for 18h (cultivation temperature 30°C, rotating speed 200rpm); put 8ml bacterial suspension in 10ml sterilized centrifuge tube, Reserved, all are made and used now.
[0023] 2. Preparation of bacterial suspensions of Escherichia coli and other pathogenic bacteria
[0024] (1) Pick a ring of bacterial lawn from the inclined surface and inoculate it into 50m1 TSB liquid medium, and culture it on a shaking table for 18h (cultivation temperature
[0025] 30 0C, speed 140rpm);
[0026] (2 Take 5m1 of the above-mentioned bacterial suspension and re-inoculate it into 50m1 of newly prepared TSB liquid medium, shake and cultivate for 18h
[0027]...
Embodiment 3
[0039] The preparation of embodiment 3 microorganism bacterial agent (bacillus licheniformis preparation)
[0040] 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 licheniformis;
[0041] 2. Use the Bacillus licheniformis liquid 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 1:0.8, and cultivate at 37°C for 24 -36 hours, wh...
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