Composite microbial preparation for fermented feed and fermented expanded pellet feed
A technology of extruded pellet feed and compound microorganisms, applied in animal feed, animal feed, application, etc., can solve the problems of poor probiotic effect and easy inactivation of probiotics, and achieve strong degradation ability, good feeding effect, and acid production capacity strong effect
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Embodiment 1
[0030] Example 1: Isolation, screening and gene sequencing of Lactobacillus salivarius LS strain
[0031] Source of strains: intestinal contents of healthy grass carp (aquaculture base of Jiangxi Agricultural University).
[0032] Method: The healthy grass carp was dissected in an ultra-clean workbench, and its intestinal contents were directly diluted 10 times in sterile saline to 10 -2 -10 -8 Concentration, take 0.1ml and spread on the MRS solid medium containing 1% calcium carbonate, and culture at 30°C for 24-48h. Pick a single colony that produces a calcilytic circle from the MRS solid medium containing 1% calcium carbonate and purify it on an ordinary MRS solid medium. After purification twice, carry out Gram staining and biochemical identification. The colony was inoculated in the corresponding liquid medium, and cultured at 30°C and 180r / min for 24-48h.
[0033] A single strain in the MRS liquid medium was planted on the MRS solid medium with 1% calcium carbonate by...
Embodiment 2
[0038] Example 2: Isolation, screening and gene sequencing of Bacillus subtilis B1 strain
[0039] Sources of strains: termite intestinal material (termites collected from the campus forest of Jiangxi Agricultural University); intestinal contents of healthy grass carp (aquaculture base of Jiangxi Agricultural University); a strain of Bacillus subtilis and a strain of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) , two strains have antibacterial effect on Aeromonas hydrophila (Aeromonas hydrophila), isolated and preserved by our laboratory.
[0040] Indicator bacteria: Aeromonas hydrophila AH-S1, Aeromonas sobria (Aeromonas sobria) AS-S2, isolated from diseased eel liver, preserved by our laboratory.
[0041] Medium:
[0042] Guaiacol primary screening medium: yeast extract 10g / L, glucose 20g / L, guaiacol 2.5g / L, agar 15g / L, adjust pH to 7.0, autoclave at 121°C for 20min.
[0043] RB brilliant blue medium: RB brilliant blue 0.03g / L, yeast extract 5g / L, tryptone 10g / L...
Embodiment 3
[0061] Embodiment 3: the composite ratio optimization of three strains of bacteria in the composite microbial preparation
[0062] Compound microbial preparations:
[0063] A strain of Bacillus subtilis (Bacillus subtilis) B1 capable of degrading lignin, isolated from termite guts, obtained through separation and screening in Example 2, with a preservation number of CGMCC NO.20405;
[0064] A Lactobacillus salivarius LS strain, isolated from the intestinal contents of healthy grass carp, was obtained by isolation and screening in Example 1, and the preservation number is CGMCC NO.14458;
[0065] A strain of Saccharomyces cerevisiae (Saccharomyces cerevisiae) J1 was purchased from the General Microorganism Center of China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC NO.2.388.
[0066] Feed formula:
[0067] Feed raw materials and processing and granulation were provided by Dayou Nong Biotechnology Co., Ltd. The test feed was extrude...
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