Bacillus subtilis preparation capable of being applicable to intestinal tract fixed implantation and improving digestion rate and immunity and preparation method thereof
A technology of Bacillus subtilis and digestibility, which is applied in the technical field of feed additives for poultry and livestock breeding, can solve problems such as excessive use of antibiotics, antibiotic residues in poultry and livestock products, hidden food safety hazards, etc., and achieve improved antioxidant capacity, good acid resistance and The effect of bile salt and good antibacterial activity
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Embodiment 1
[0023] Embodiment 1: the separation and identification of this Bacillus subtilis:
[0024] Collect bacterial cells: Take 1 ml of the mixed bacterial liquid from fermented plants, centrifuge at 3500 r / min for 10 min in a centrifuge, collect the supernatant in another clean centrifuge tube, discard the precipitate; put the collected supernatant in Centrifuge at 10,000 r / min for 10 min in a high-speed centrifuge, collect the precipitated bacteria, and discard the supernatant.
[0025] Collect spores: Add 1 ml of sterile water to the centrifuge tube used to collect bacteria, mix well on a vortex shaker, place in a constant temperature water bath at 80°C for 15 minutes, kill vegetative cells and other bacterial cells, and collect spores.
[0026] Spore recovery: Inoculate an appropriate amount of spore collection liquid into LB liquid medium in a clean bench, place on a constant temperature shaker at 37 °C and 180 r / min for enrichment and culture for 24-36 hours, and then take 1 ml...
Embodiment 2
[0043] Example 2: Cultivation and enrichment of Bacillus subtilis: The purified strain was added to LB liquid medium (per liter of water: tryptone 10g, yeast extract 5g, NaCl 10g pH 7.0) at a ratio of 1:100. 37°C, 180 r / min placed on a constant temperature shaker for enrichment culture for 24-36 h.
Embodiment 3
[0044] Example 3: Antioxidant activity detection of Bacillus subtilis on Drosophila
[0045] 1. Test materials: w1118 wild-type Drosophila; the Bacillus subtilis strain of the present invention, the composite bacterial solution of Bacillus pumilus (513-A) and Bacillus gobiensis (513-O) preserved in our laboratory.
[0046] 2. Preparation of Drosophila basal medium: Liquid A: Weigh 10g of agar powder, 70g of edible white sugar, add to 500ml of distilled water and boil. Liquid B: Weigh 85g of corn flour, add it to 500ml of distilled water and boil.
[0047] Slowly pour liquid B into liquid A, stir while heating, cool slightly after forming a paste, add 18g yeast powder to the mixture, then stir evenly, then add 5ml propionic acid, mix evenly, divide into sterilized containers In Drosophila culture tubes (3×10 cm), add 5 ml of medium to each tube, seal with a cotton plug, and store at 4°C for future use.
[0048] 3. Preparation of Drosophila culture medium containing Bacillus s...
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