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Preparation and use of bacillus subtilis and clostridium butyricum composite bacterial preparation

A technology of Bacillus subtilis and Clostridium butyricum, applied in the direction of application, bacteria, and microbial-based methods, can solve the problems of reducing equipment utilization, difficulty in cultivating, and increasing energy consumption, so as to promote colonization, enhance the effect, The effect of strong competition

Active Publication Date: 2011-10-19
山东亚太海华生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the fermentation medium of the three bacteria needs to be sterilized separately, which increases energy consumption and reduces the utilization rate of equipment
[0005] Although the simultaneous culture of Bacillus subtilis and Clostridium butyricum is difficult due to the difference in oxygen requirements, this contradictory relationship provides the basis for the synergistic effect of the two in vivo

Method used

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  • Preparation and use of bacillus subtilis and clostridium butyricum composite bacterial preparation
  • Preparation and use of bacillus subtilis and clostridium butyricum composite bacterial preparation
  • Preparation and use of bacillus subtilis and clostridium butyricum composite bacterial preparation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Prepare the seed culture solutions of Bacillus subtilis and Clostridium butyricum respectively, in which the concentration of the bacteria solution is 5.0×10 8 CFU / ml.

[0026] 2. Make a joint medium, specifically: high-temperature bean cake powder 1.8%, urea 0.1%, corn steep liquor 0.1%, corn starch 0.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.1%, sulfuric acid Manganese 0.01%.

[0027] 3. Add the Bacillus subtilis seed liquid in step 1 to the joint culture medium at a ratio of 2%, and use 10% dilute hydrochloric acid or 10% sodium hydroxide to automatically adjust the pH value to 7.0, ventilate, stir, and control dissolved oxygen 20%, cultured at 37°C for 20 hours, then stopped aeration, and continued stirring for 2 hours.

[0028] 4. Add the Clostridium butyricum seed solution in step 1 to the joint medium at a ratio of 2% using a peristaltic pump, and culture at 35°C for 28 hours. Sampling was taken to det...

Embodiment 2

[0031] 1. Prepare the seed culture solutions of Bacillus subtilis and Clostridium butyricum respectively, and the concentration of the bacteria solution is 1.0×10 8 CFU / ml.

[0032] 2. Make a joint medium, specifically: high-temperature bean cake powder 2.5%, urea 0.2%, corn steep liquor 0.3%, corn starch 0.7%, dipotassium hydrogen phosphate 0.3%, potassium dihydrogen phosphate 0.3%, magnesium sulfate 0.2%, sulfuric acid Manganese 0.02%.

[0033] 3. Add the Bacillus subtilis seed liquid in step 1 to the joint culture medium at a ratio of 2%, use 10% dilute hydrochloric acid or 10% sodium hydroxide to automatically adjust the pH value to 6.8, ventilate, stir, and control dissolved oxygen 30%, cultured at 37°C for 20 hours, then stopped aeration, and continued stirring for 2 hours.

[0034] 4. Add the Clostridium butyricum seed solution in step 1 to the joint medium at a ratio of 2% using a peristaltic pump, and culture at 35°C for 26 hours. Sampling was taken to detect the t...

Embodiment 3

[0037] 1. Prepare the seed culture solutions of Bacillus subtilis and Clostridium butyricum respectively, and the concentration of the bacteria solution is 0.5×10 8 CFU / ml.

[0038] 2. Make a joint culture medium, specifically: high-temperature bean cake powder 3.5%, urea 0.3%, corn steep liquor 0.5%, corn starch 1.0%, dipotassium hydrogen phosphate 0.5%, potassium dihydrogen phosphate 0.5%, magnesium sulfate 0.3%, sulfuric acid Manganese 0.05%.

[0039] 3. Add the Bacillus subtilis seed liquid in step 1 to the joint culture medium at a ratio of 5%, and use 10% dilute hydrochloric acid or 10% sodium hydroxide to automatically adjust the pH value to 6.5, ventilate, stir, and control dissolved oxygen 20%, cultured at 37°C for 18 hours, then stopped aeration, and continued stirring for 2 hours.

[0040] 4. Add the Clostridium butyricum seed liquid in step 1 to the joint medium at a ratio of 2% using a peristaltic pump, and culture at 35°C for 24 hours. Sampling was carried out...

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Abstract

The invention discloses a method for preparing a bacillus subtilis and clostridium butyricum composite bacterial preparation, which is characterized by comprising the following steps: (1) preparing seed culture solution, namely preparing a seed solution containing clostridium butyricum and a seed solution containing bacillus subtilis respectively, wherein the concentration of the bacterial solution is 0.5*10<8> to 1.0*10<8>CFU / ml; (2) preparing a coculture medium; (3) adding the seed solution of the bacillus subtilis into the coculture medium prepared by the step (1) according to a ratio of 2 percent, automatically regulating the pH value to 6.5 to 7.0 by using 10-percent diluted hydrochloric acid or 10-percent sodium hydroxide, introducing gas, stirring, controlling dissolve oxygen content to be 20 to 50 percent, culturing for 18 to 20 hours at 37 DEG C, stopping introducing gas, and continuing to stir and culture for 2 to 4 hours; and (4) adding the seed solution of the clostridium butyricum prepared by the step (1) into the coculutre medium by using a peristaltic pump according a ratio of 2 percent, standing at 35 DEG C and culturing for 24 to 28 hours to obtain the bacillus subtilis and clostridium butyricum composite bacterial preparation. In the invention, a microecological preparation is mixed with feed according to a ratio of 0.5 percent, the feed is directly used to feed livestock, the diarrhea rate and death rate of the livestock are lowered effectively, yield is increased, and medicine administration cost is saved.

Description

technical field [0001] The present invention relates to the preparation of the compound bacterial preparation of Bacillus subtilis and Clostridium butyricum and its application in livestock and poultry breeding, specifically relates to a kind of two strains of Bacillus subtilis and Clostridium butyricum as raw materials, which are used in The synergistic relationship in the fermentation process, the joint preparation method of obtaining the compound probiotic preparation of Bacillus subtilis and Clostridium butyricum by one-time fermentation and its application in preventing livestock and poultry diarrhea. Background technique [0002] Bacillus subtilis is a gram-positive aerobic bacterium, which rapidly consumes free oxygen in the environment during the growth process, causing intestinal hypoxia, promoting the growth of beneficial anaerobic bacteria, and producing organic acids such as lactic acid, reducing the pH value of the intestinal tract , Indirectly inhibit the growt...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K1/16C12R1/125C12R1/145A23K10/18
Inventor 位宾张杰丛晓燕朱小玲
Owner 山东亚太海华生物科技有限公司
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