Manure enterococcin strain and use thereof
A technology of Enterococcus faecium and strains, applied in bacteria, applications, animal feed, etc., can solve the problems of low viable bacteria count and inability to adsorb, and achieve the effects of fast growth, convenient cultivation and strong pertinence
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Embodiment 1
[0030] Embodiment 1, strain isolation and screening
[0031] Make a suspension of the intestinal contents of healthy pigs, enrich and culture them in nutrient broth for 24 hours (35-38°C), put them in an 80°C water bath and heat them for 5-10 minutes to kill bacteria that are not resistant to high temperatures . Take it out and let it stand for 10 minutes, spread the sample solution evenly with a sterile spatula after dilution, cover all the plates with melted SBA medium, and incubate in a biochemical incubator at 35-38°C for 48 hours. Pick iron-red colonies and dilute them slightly and observe under the microscope. For colonies whose cells are elliptical or club-shaped, extend in the direction of connection, have a diameter of 0.5-1.0 μm, and form pairs or short chains, continue to screen.
[0032] The catalase-negative and xylose-arginine dihydrolase-negative strains were further separated and purified by streaking, and the well-growing colonies were selected according to t...
Embodiment 2
[0033] Embodiment 2, strain identification
[0034] (1) Morphological characteristics
[0035]Strain ZD-0401 is Gram-positive, oval or club-shaped, without spores, and without flagella. It can grow on nutrient agar and MacConkey agar, and appears small (less than 1mm) smooth and round colonies. On the blood agar plate, a slightly larger (about 1mm) colony is formed, which is smooth, moist and easy to emulsify. It grows uniformly and turbidly in liquid medium, and it is also easy to form long chains.
[0036] (2) Biochemical properties
[0037] Strain ZD-0401 was insensitive to gentamicin and streptomycin, but sensitive to penicillin, fluoromycin, erythromycin, tetracycline, rifampicin, vancomycin and chloramphenicol. Can utilize (positive) sucrose, melibiose, glucose (gas production), lactose, maltose and mannitol; cannot utilize (negative) rhamnose, raffinose, sorbitol and melezitose.
[0038] (3) Culture characteristics
[0039] Strain ZD-0401 can grow well at a tempera...
Embodiment 3
[0040] Embodiment 3, making probiotics
[0041] All "%" in this example represent a weight ratio.
[0042] Method 1, liquid fermentation method:
[0043] Slant medium: nutrient agar medium and 10% bran extract, 2% agar, natural pH, autoclaved at 121°C for 30min.
[0044] Composition of nutrient salt solution (%): KH 2 PO 4 , 0.05; MgSO 4 .7H 2 O, 0.05; CaCl 2 , 0.01; NaCl, 0.10; FeSO 4 .7H 2 O, 0.005; MnSO 4 .H 2 O, 0.001; ZnSO 4 .7H 2 O, 0.0005; CoCl 2 , 0.0005; the rest is water.
[0045] Culture medium: Add 5% bran extract and 1% yeast extract to the nutrient salt solution to adjust the pH to 6.5.
[0046] Culture conditions: Fill 100ml of medium into a 250ml Erlenmeyer flask, inoculate 1.0ml of Enterococcus faecium ZD-0401 suspension, shake at 100r / min, and incubate at 37°C for 36h.
[0047] Method 2, solid fermentation method:
[0048] (1), first-class seeds (test tube slant seeds): inoculate the original strain into the test tube nutrient agar solid mediu...
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