Bacillus subtilis and application thereof in biological feed additives
A Bacillus subtilis, biological feed technology, applied in animal feed, animal feed, microorganism-based methods, etc., can solve the problem of poor resistance to the physiological environment of the digestive tract, poor resistance to feed processing, unreliable safety, and insufficient bacteriostatic effect. Significant other problems, to achieve the effect of promoting intestinal microecological balance, no drug withdrawal period, increasing weight gain rate and feed conversion rate
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Embodiment 1
[0029] Primary screening and separation and purification of bacterial classification of embodiment 1
[0030] Slaughter healthy suckling pigs, respectively take a certain amount of the contents of the stomach, duodenum, jejunum, ileum, cecum and colon near the mucosa in sterile water, stir and make 10-fold dilution, inoculate in BPY liquid medium, in Incubate at 200 rpm in a constant temperature shaker at 35°C for 30 hours.
[0031] Boil the above-mentioned bacterial solution in boiling water for 10 minutes, then streak culture on the BPY medium plate, pick out various colonies with different shapes, streak culture and separate them, and check under the microscope whether there are spores and whether they are single colonies. Inoculate the isolated 40 single colonies and spores into the slant of BPY medium test tubes at 35°C for 30 hours, and store them in the refrigerator at 4°C for later use.
[0032]The composition of BPY medium is as follows: beef extract 0.5g, peptone 1g...
Embodiment 2
[0033] Example 2 Enzyme Production Ability Screening
[0034] The 40 strains of spores screened out in Example 1 were screened for excellent strains with high protease production using casein hydrolysis plates. Observe the size of the transparent circle of 40 strains of spores, and select 5 strains of spores with large transparent circles (that is, higher protease activity) to enter the next step of screening.
[0035] Casein plate medium: beef extract 8g, yeast powder 2g, peptone 5g, sodium chloride 2g, casein 4g, agar 20g, distilled water 1000ml, pH7.2~7.4.
Embodiment 3
[0036] Example 3 Screening of Anti-digestive Tract Physiological Environment Excellent Bacterial Strains
[0037] The 5 bacterial strains screened out in Example 2 were screened for excellent strains resistant to digestive tract physiological environment.
[0038] (1) The preserved slant strains were inoculated in a test tube containing 10ml of BPY liquid medium, cultured at 35°C for 30h, and then inoculated into 9.0ml of artificial simulated gastric juice with pH values of 2.0, 3.0, and 4.0 at 0h Count as a control, 2h, 6h sampling with PBS buffer (pH7.0, 10mmol / L) by 10-fold serial dilution, carry out plate live bacteria technology, calculate the survival rate, and screen the excellent strains with a survival rate of more than 3%.
[0039] Preparation of artificial simulated gastric juice: measure 16.4ml of concentrated hydrochloric acid, add distilled water to 1000ml, make basic artificial gastric juice, adjust pH value to 2.0, 3.0, 4.0 with hydrochloric acid or sodium hy...
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