A strain of Bacillus amyloliquefaciens and its application, aquatic feed and aquaculture method
A technology for dissolving starch spores and aquatic feed, which is applied in the directions of biochemical equipment and methods, applications, animal feed, etc., to protect the immune system of the body, improve the disease resistance of fish, and improve the utilization efficiency.
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Embodiment 1
[0034] This example provides a strain of Bacillus amyloliquefaciens SS1 (hereinafter referred to as "SS1"), which is deposited in the China Center for Type Culture Collection, the preservation number is: CCTCC M 2020024, and the preservation date is: January 2020 7th.
[0035] The above strain information is as follows:
[0036] 1. The source of the strain
[0037] This strain was isolated from the gut of healthy tilapia (Oreochromis mossambicus).
[0038] 2. Morphological characteristics
[0039] Gram-positive, such as figure 1 As shown, the colonies that were cultured upside down on LB medium plate at 28°C for 12 h were white irregular circles with dry surface and volcano-like white bumps in the center of the colonies, similar to fried eggs.
[0040] 3. Enzyme production characteristics
[0041] 1) Dip a single colony of SS1 with a toothpick, place it on the starch medium, invert at 28°C for 12 hours, drop Lugol's iodine solution on the culture plate, and let it stand f...
Embodiment 2
[0067] The present embodiment provides the separation method of Bacillus amyloliquefaciens SS1 (hereinafter "SS1") described in Example 1, which comprises the following steps:
[0068] S1. According to tryptone 5g / L, yeast extract 2g / L, KH 2 PO 4 2 g / L, MgSO 4 ·7H 2 0 2g / L, NaCl5g / L, soluble starch 10g / L, agarose 20g / L ratio preparation starch culture medium, after autoclaving, pour the plate in the ultra-clean bench to obtain the starch culture medium solid plate;
[0069] S2. Mix the intestinal contents of fish (such as tilapia) with PBS buffer at a ratio of 1:9 (w / v, g / ml), centrifuge at 1500 rpm / min, and take 100 μL of the supernatant for coating be distributed on the solid plate of the starch medium;
[0070] S3. After culturing upside down at 28°C for 24 hours, screen the colonies according to the size, shape and color of the colonies, and slant the screened colonies for preservation and numbering. At the same time, carry out 16S rRNA molecular identification on the ...
Embodiment 3
[0072] The present embodiment provides a method for aquaculture using the Bacillus amyloliquefaciens SS1 (hereinafter referred to as "SS1") described in Example 1, which specifically includes the following steps:
[0073] S1. Temporary cultivation of fry: buy 600 fish fry (such as tilapia fry), temporarily raise them with commercial feed for 2 weeks, and then put them into aerated water at 28°C for temporary cultivation. Change the water every two days until the fry are adapted to the growth environment. After the disease, the temporary culture was terminated; the fry were divided into five groups, normal group (CON), high starch group (HCD), probiotic group 1 (HCB1), probiotic group 2 (HCB3), and probiotic group 3 (HCB3). Three parallel groups, each with 30 tails, all weigh 1.6±0.1g;
[0074] S2 and SS1 bacterial liquid preparation: take the above-mentioned Bacillus amyloliquefaciens SS1 at 1:1000 (w / v, g / ml) and inoculate it into LB liquid medium for recovery, shake the bact...
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