Clostridium butyricum used for feeding, and application thereof
A technology of Clostridium butyricum and bacteria agent, applied in the direction of application, bacteria, animal feed, etc., to achieve broad application prospects, strong inhibitory effect, and the effect of enhancing animal disease resistance
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Embodiment 1
[0029] Example 1. Isolation, identification and preservation of Clostridium butyricum B1 CGMCC No. 4845
[0030] 1. Isolation of Clostridium butyricum B1 CGMCC No. 4845
[0031] Clostridium proliferation medium (RCM medium): yeast extract 3g, beef extract 10g, tryptone 10g, glucose 5g, soluble starch 1g, sodium chloride 5g, sodium acetate trihydrate 3g, cysteine hydrochloride 0.5g, 0.5% methylene blue 0.2mL, distilled water 1000mL; adjust the pH to 7.1±0.1 before use, cover the culture medium with 5mm liquid paraffin, and sterilize it for use at 115°C for 20 minutes.
[0032] Clostridium selective medium (TSN medium): tryptone 15g, yeast extract 10g, sodium sulfite 1g, iron citrate 0.5g, novobiocin 0.02g, polymyxin 0.05g, agar 0.5g (with solid Used for culture medium), pH 7.2, 115°C, 20min sterilization for later use.
[0033] Multi-point and mixed collection of cellar mud from Hubei Jinpai Wine Co., Ltd. Liuren Bazhen base wine base wine cellar, Hubei Zhijiang Wine Co., Ltd. wine ...
Embodiment 2
[0042] Example 2: Analysis of acid resistance properties of Clostridium butyricum B1 CGMCC No. 4845 spores
[0043] Strain activation, inoculate Clostridium butyricum B1 CGMCC No. 4845 into a test tube containing 9mL RCM medium at an inoculum of 1% (volume / volume percentage), and culture at 37°C for 48h under anaerobic static. Centrifuge and discard the supernatant, wash and suspend the bacteria. Take 0.2mL centrifugal washing bacteria suspension and inoculate respectively 2mL pH 1.0, pH 2.0, pH 3.0 sterile phosphate buffer solution (A solution: phosphoric acid solution, B solution: disodium hydrogen phosphate solution; by adjusting A solution, B solution Proportion, the pH was adjusted to 1.0, 2.0, 3.0) in an anaerobic test tube, placed in a 37°C incubator, and counted viable bacteria at 0h, 1h, 2h, and 3h to calculate the survival rate.
[0044] Results The spore survival rate was 60% after the phosphate buffer with pH value of 1 for 1 hour, and 32.7% after the phosphate buffer ...
Embodiment 3
[0045] Example 3 Analysis of the bile salt tolerance properties of Clostridium butyricum B1 CGMCC No. 4845
[0046] The strain is activated, and Clostridium butyricum B1 CGMCC No. 4845 is inoculated into a test tube containing 9mL RCM medium at an inoculum of 1% (volume / volume percentage), and cultured at 37°C for 48h under anaerobic static. Centrifuge to remove the supernatant, wash and suspend the bacteria. Take 0.2mL centrifugal washing bacteria suspension and inoculate it in 2mL RCM liquid medium containing 0.1%, 0.3%, 0.5% (mass percentage concentration) of porcine bile salt. Placed in a 37°C incubator, counted viable bacteria at 0h, 1h, 2h, 3h, and 20h, and calculated the survival rate.
[0047] The test results showed that the survival rate of the strain was 76.7% after 1h in a bile salt solution of 0.5% (mass percentage concentration) and 24.3% after 20h; in a bile salt solution of less than 0.3% (mass percentage concentration) for 20h The post-survival rate was 66.5%, an...
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