Bacillus coagulans and application thereof
A technology of Bacillus coagulans and uses, applied in the field of microorganisms, can solve problems such as few applications, and achieve the effects of simple environment, promotion of fish growth and development, and high production and application value.
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Embodiment 1
[0034] Embodiment 1, isolation and screening of probiotic strains
[0035] Put 5g of coffee powder in 200ml of water, heat at 80°C for 10min, and filter to obtain coffee grounds.
[0036] Add 50 ml of purified water to the above 5 g of coffee powder corresponding to the obtained coffee grounds, ferment at 50° C. for 24 hours, and take the supernatant.
[0037] Use a pipette with a sterile tip to pipette 5ml of the supernatant into a 10ml sterile disposable pipette and cap it. Dilute to an appropriate concentration in an ultra-clean bench (dilute 5ml of the supernatant to 50ml with purified water), and use a 200μl pipette to draw 100μl of the diluted solution into the MRS-CaCO 3 Incubate in an agar dish at 50°C for 24-48h. After the colony was formed, the colony was picked with an inoculation loop, inoculated in MRS liquid medium, and cultured at 50°C for 24 hours. After the strain grows well, it is streaked and inoculated on the MRS agar medium again, and placed in the same...
Embodiment 2
[0041] Embodiment 2, the identification of probiotic strain
[0042]The isolated and purified Bacillus coagulans (Bacillus coagulans) XY-4 was preliminarily identified as Bacillus coagulans according to the 16S rDNA sequencing identification method. Afterwards, according to the comprehensive analysis of the experimental data such as the cell morphology, physiological and biochemical characteristics, and 16S rRNA gene sequence of the strain, referring to "Bergey's Systematic Bacteriology Handbook" and related research papers in the International Journal of Systematic and Evolutionary Microbiology, the identification result is: Bacillus coagulans coagulans bacilli. Its microbiological characteristics are shown in Table 1 above.
[0043] The preservation information of bacterial strain of the present invention is as follows:
[0044] Strain XY-4, the preservation name is Bacillus coagulans (Bacillus coagulans), preservation unit: General Microbiology Center of China Microbiolog...
Embodiment 3
[0045] Embodiment 3, the preparation of Bacillus coagulans (Bacillus coagulans) XY-4 effective thallus:
[0046] Take Bacillus coagulans (Bacillus coagulans) XY-4 out of -80°C, place it in MRS agar medium at 50°C for 18h to activate the first generation, and pick a single colony of an inoculation loop into 5ml of MRS liquid medium 50°C static culture for 18h to reactivate, take the second activated liquid culture and add 2% (volume %) to the final liquid medium (that is, new MRS liquid medium), continue to stand at 50°C Cultivate for 18h. The final culture was placed in a sterile centrifuge tube and centrifuged (12000 RPM for 1 minute). Discard the supernatant, wash twice with pisciculture water to obtain pure bacteria, which need to be re-dissolved in pisciculture water and diluted to the optimum concentration before use.
[0047] Fish culture water is the water that meets the recommended conditions of the National Zebrafish Resource Center (CZRC).
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