Feed additive, premix and mixture containing like-bacteriocin and pediococcus acidilactici
A technology of Pediococcus lactis and feed additives, applied in the direction of bacteria, animal feed, animal feed, etc., can solve the problems of not being able to completely get rid of the dependence on antibiotic growth promoters, not being able to fully mobilize probiotics, and unsatisfactory use. Improve the intestinal microbial environment, prevent diarrhea, and eliminate drug residues
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Embodiment 1
[0033] 1) Preparation of Bacillus subtilis fermentation broth
[0034] MNB medium was used for fermentation at 30°C for 20 hours, the initial pH was 7, the inoculum size was 3%, and the shaker speed was 225rpm.
[0035] The formula of MNB medium is: glucose, 5.0g / L; peptone, 5.0g / L; meat extract, 3.0g / L; yeast extract powder, 1.0g / L; MgSO 4 7H2O, 0.5g / L; MnSO 4 ·H 2 O, 0.005g / L; pH, 7.0.
[0036] 2) Extraction of bacteriocinoids by ammonium sulfate salting-out method
[0037] Prepare the sterile filtrate from the cultivated Bacillus subtilis bacteria liquid, slowly add the ground ammonium sulfate powder into the sterile filtrate, and stir gently to ensure that the ammonium sulfate is completely dissolved, after stirring slowly for 1 hour, place it in a refrigerator at 4°C . Ammonium sulfate gradient treatment was 70%. After overnight, the solutions with different saturations were taken out and centrifuged separately (10000×g, 30min, 4°C) to collect the precipitate.
[0...
Embodiment 2
[0047] The preparation of embodiment 2 Pediococcus lactis bacteria powder
[0048] 1) Preparation of Pediococcus lactis CGMCC No.6500 seed solution
[0049] Inoculate the strains preserved on the slant of the test tube into an anaerobic tube containing 20mL of MRS liquid medium, and incubate at a constant temperature for 20 hours at 37°C until the number of viable bacteria reaches 10 7 CFU / mL, inoculate at 1% in a 250mL Erlenmeyer flask containing 100mL MRS liquid medium, culture anaerobically at 37°C for 20h, and wait for the number of viable bacteria to reach 107CFU / mL (MRS agar medium plate count) or more, as seeds liquid for use.
[0050] 2) Liquid submerged fermentation culture
[0051] The above-mentioned Pediococcus lactis liquid seed solution was added to a 50L fermenter with an inoculation amount of 1%. The culture parameters were: temperature, 37°C; stirring speed, 50rmp; pH value, 6.0±0.5; culture time, 20h. During the fermentation process, 0.5% of the total ferm...
Embodiment 3
[0059] Preparation of Example 3 Bacteriocins and Pediococcus lactis Mixed Feed Additive
[0060] Prepared by uniformly mixing 50% (wt) of the Bacillus bacteriocin dry powder prepared in Example 1 with 50% (wt) of the Pediococcus lactis powder prepared in Example 2. Its quality standards are: bacteriocinoid dry powder 102mg / g, viable lactic acid bacteria 12 billion / g, moisture ≤ 10%, arsenic ≤ 0.5mg / kg, lead ≤ 1.0mg / kg.
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