Preparation technology of bacillus feed additive agent
A technology for feed additives and Bacillus, which is applied in the field of preparation of Bacillus feed additives, can solve the problems of limiting the wide application of microecological preparations, affecting the application effect, high product price, etc., and achieves good inhibition of pathogenic bacteria, stable application effect and good resistance. Inverse performance effect
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Embodiment 1
[0020] 1) inoculating the strains of Bacillus licheniformis into BPY seed medium for activation, culturing at 200 rpm at 37°C for 18 hours to obtain a culture with a spore rate above 95%;
[0021] 2) Determination of acid resistance: inoculate the cultures prepared above at 5% inoculum size into artificial simulated gastric juices with pH values of 2.0, 3.0, and 4.0, count at 0h as a control, and use phosphate buffer for sampling at 2h and 6h in a 10-fold series Dilute, count the viable bacteria on the plate, and calculate the survival rate.
[0022] Preparation of artificially simulated gastric juice: measure 16.4 ml of 9.5%-10.5% concentrated hydrochloric acid, add distilled water to 1000 ml, make basic artificial gastric juice, adjust pH value to 2.0, 3.0, 4.0 with hydrochloric acid or sodium hydroxide, take 10 mL (9 mL) each , divided into test tubes, steam sterilized at 100°C for 15 minutes, under sterile conditions, add 0.100g of pepsin to every 10mL of liquid.
[002...
Embodiment 2
[0033] 1) Cultivation of Bacillus licheniformis, primary culture: Inoculate the strains on the BPY plate medium, cultivate for 24 hours at 30°C, rejuvenate the Bacillus licheniformis, and form a single colony, pick a single colony and inoculate it on the slant medium, Cultivate at 30°C for 6 hours; Secondary cultivation: Inoculate Bacillus licheniformis on the first-grade BPY slant medium, and cultivate it at 30°C for about 16 hours to make it in the middle and late logarithmic period, and obtain first-grade seeds; Third-grade liquid cultivation: First-grade The first-grade seeds are made of bacterial suspension with sterile water, inoculated into BPY seed medium, the temperature is 30°C, the rotation speed is 200rpm, the tank pressure is 0.05Mpa, the ventilation ratio: 1:0.6, and cultivated for 14h to obtain the second-grade seed liquid and the seed liquid ; 2) liquid fermentation culture of Bacillus licheniformis, the inoculum size of the seed liquid is 3%, and the fermentati...
Embodiment 3
[0035] 1) Cultivation of Bacillus licheniformis, primary culture: inoculate the strains on BPY plate medium, culture at 37°C for 12 hours, rejuvenate Bacillus licheniformis and form a single colony, pick a single colony and inoculate it on the slant medium , cultured at 37°C for 24 hours; secondary culture: inoculate Bacillus licheniformis on the first-grade BPY slant medium, and cultivate it at 37°C for 12 hours to make it in the late logarithmic period, and obtain first-grade seeds; third-grade liquid culture: a The first-grade seeds are made of bacterial suspension with sterile water, inoculated into the BPY seed medium, the temperature is 37°C, the rotation speed is 250rpm, the tank pressure is 0.05Mpa, the ventilation ratio: 1:0.8, and cultivated for 10h to obtain the second-grade seed liquid; 2) For the liquid fermentation culture of Bacillus licheniformis, the inoculation amount of the seed liquid is 10%, and the fermentation is carried out until the number of bacteria r...
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