A strain of Bacillus subtilis capable of inhibiting Clostridium perfringens and its application
A technology for Clostridium perfringens and Bacillus subtilis, which is applied to medical preparations containing active ingredients, bacteria, antibacterial drugs, etc., can solve the problems of large differences in the probiotic properties of Bacillus, and improve daily weight gain. , the effect of reducing mortality and reducing injuries
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Embodiment 1
[0030] Example 1: Isolation and identification of Bacillus subtilis with the effect of inhibiting Clostridium perfringens
[0031] 1 Materials and methods
[0032] 1.1 Materials
[0033] 1.1.1 Separation of sources
[0034] Branches, roots, leaves, bark, etc. of Eucommia ulmoides in Tai'an and surrounding areas.
[0035] 1.1.2 Isolation medium
[0036] LB solid medium: 0.5% of yeast extract, 1% of peptone, 1% of NaCl, 1.5% of agar powder, and the pH value is 7.0; calculated by mass percentage.
[0037] 1.1.3 Incline medium
[0038] Separation medium with 1.1.2;
[0039] 1.1.4 Fermentation medium
[0040]The fermentation medium is LB liquid medium, and its composition is: 0.5% of yeast extract, 1% of peptone, 1% of NaCl, and the pH value is 7.0; it is calculated by mass percentage.
[0041] 1.1.5 Medium for indicator bacteria
[0042] Escherichia coli and Staphylococcus aureus were cultured using LB medium, and the medium formula was: 0.5% yeast extract, 1% peptone, 1% ...
Embodiment 2
[0096] Embodiment 2: the preparation technology of probiotics
[0097] 1 material
[0098] 1.1 Strains: The strain BLCC1-0155 is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCCNO:M 2015278.
[0099] 1.2 Medium:
[0100] The formula of the seed liquid medium is: glucose 20g / L, peptone 10g / L, yeast extract 5g / L, adjust the pH to 7.0, and sterilize at 115°C for 20min;
[0101] The solid slant medium is the agar powder with a mass fraction of 1.5% added to the above-mentioned seed liquid medium;
[0102] The liquid fermentation medium formula is: glucose 20g / L, peptone 10g / L, yeast extract 5g / L, sodium chloride 5g / L, pH7.0.
[0103] 2 methods
[0104] 2.1 Preparation process of bacteria powder
[0105] (i) Bacteria: Bacillus subtilis BLCC1-0155, CCTCC NO: M2015278;
[0106] (ii) Slant culture: inoculate the lyophilized powder strain of Bacillus subtilis BLCC1-0155 on a solid slant medium, and culture at 37°C for 24 hours;
[0...
Embodiment 3
[0114] Embodiment 3: the establishment of chicken necrotic enteritis model
[0115] 1 Materials and methods
[0116] 1.1 Materials
[0117] AA broilers were purchased from Shandong Taian Daewoo Breeding Farm; 510 feed was purchased from Shandong Liuhe Group Co., Ltd. Clostridium perfringens type A CVCC49 was purchased from China Veterinary Microbiology Culture Collection Center.
[0118] 1.2 Method
[0119] Select 50 1-day-old AA broilers and randomly divide them into 5 groups, which are control group (no challenge), challenge 1 group (according to Clostridium perfringens 10 9 The dose of cfu / only to challenge the virus), challenge the poison 2 groups (according to Clostridium perfringens 10 8 The dose of cfu / only to challenge the poison), three groups of the challenge (according to Clostridium perfringens 10 7 The dose of cfu / only to challenge the virus), challenge 4 groups (according to Clostridium perfringens 10 6 The dose of cfu / only to attack poison). The chicks we...
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