A strain of Bacillus licheniformis fa6 that can improve the growth performance and immunity of grass carp and regulate the structure of intestinal flora and its application
A technology of Bacillus licheniformis and regulating intestinal flora, applied in the direction of application, microbe-based methods, bacteria, etc., to achieve strong tolerance, increase the length of intestinal villi, and improve the effect of non-specific immunity and antioxidant function
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Embodiment 1
[0041] The isolation and screening of embodiment 1 bacillus licheniformis FA6
[0042] The isolation and screening method of the above-mentioned bacillus licheniformis FA6 of the present invention mainly comprises the following steps:
[0043] (1) Sample collection
[0044] Grass carp samples (weight 536.9g) were collected from the Huanggang section of the middle reaches of the Yangtze River. The intestines of the grass carp were emptied and brought back to the experimental fish room for two months. During this period, no feed was fed and the water was changed once a week until the grass carp intestines were emptied. The flora tends to be stable. Put the fish used in the experiment on the dissection plate, wipe the body surface with 75% alcohol for 3 times, and cut it in an arc along the anus with dissecting scissors, and divide the grass carp intestine into foregut, The midgut and hindgut were ligated with a thin thread, the intestine was removed, the fat on the outer wall ...
Embodiment 2
[0053] Molecular Identification of Example 2 Bacillus licheniformis FA6
[0054] Use the bacterial universal primers to carry out PCR identification on the screened bacterial strains: extract the template DNA according to the operating instructions of the bacterial DNA extraction kit. Using the upstream primer 27F of the 16S rRNA conservative sequence: 5'-AGAGTTTGATCMTGGCTCAG-3' (as shown in SEQ ID No.2) and the downstream primer 1492R: 5'-TACGGYTACCTTGTTACGACTT-3' (as shown in SEQ ID No.3), to Bacillus licheniformis The 16s rRNA gene fragment of FA6 was amplified by PCR, and the amplified product was recovered with an agarose gel purification kit, and the recovered product was ligated and transformed into DH5α competent cells with pMD-18T vector, and spread on 57mg / ml ampicillin Incubate on penicillin LB medium at 37°C for 10 h. Randomly select transformants, use carrier universal primers M13R: 5'-CAGGAAACAGCTATGACC-3' (as shown in SEQ ID No.4) and M13F: 5'-TGTAAAACGACGGCCAG...
Embodiment 3
[0055] The carbon source utilization identification of embodiment 3 bacillus licheniformis FA6
[0056] 1 Test materials and methods
[0057] 1.1.1 Strains
[0058] Bacillus licheniformis FA6
[0059] 1.1.2 Experimental materials
[0060] API 50 CH identification system
[0061] 1.2 Test method
[0062] Inoculate Bacillus licheniformis FA6 into the API 50 CH identification system, and after incubation for 24 hours, use the software of the API 50 CH identification system to read the results.
[0063] 2.1 Test results
[0064] Table 1 Identification results of carbon source utilization of Bacillus licheniformis FA6
[0065]
[0066] +: Positive, -: Negative, w: Weak positive
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