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Bacillus cereus as well as preparation and application thereof

A Bacillus cereus and preparation technology, applied in the field of microorganisms, can solve problems such as low oxytetracycline efficiency, and achieve the effects of shortening oxytetracycline degradation time, high degradation efficiency and high selection efficiency

Inactive Publication Date: 2015-06-10
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above defects or improvement needs of the prior art, the present invention provides a highly efficient oxytetracycline-degrading bacterium, the purpose of which is to degrade oxytetracycline, thus solving the technical problem of low efficiency of degradation of oxytetracycline by ordinary composting methods without adding bacteria

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  • Bacillus cereus as well as preparation and application thereof
  • Bacillus cereus as well as preparation and application thereof
  • Bacillus cereus as well as preparation and application thereof

Examples

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preparation example Construction

[0054] The preparation method of bacillus cereus preparation provided by the invention comprises the following steps:

[0055] (a) Put the Bacillus cereus seed liquid into the culture medium of the fermenter and cultivate it, the inoculum amount is 2%-5%, until the spore formation is no less than 90%, and the fermentation liquid is obtained.

[0056] The configuration of the fermentor culture medium is as follows: soybean meal 6-7%, corn flour 7-8%, bran 1%-2%, humic acid 0.5%-1%, NaH 2 PO 4 0.05%~0.1%, K 2 HPO 4 0.05%~0.1%, (NH 4 ) 2 SO 4 0.1% to 0.2%, adjust the pH value to 7.0 to 7.4, and sterilize at high temperature and high pressure for 15 to 30 minutes.

[0057] The Bacillus cereus seed liquid is prepared by multistage cultivation with the Bacillus cereus strain, and the specific method is as follows:

[0058] (I) First-class seeds: Inoculate the strains in a 500mL Erlenmeyer flask, the liquid volume is 150mL, the inoculum volume is 4-5%, 35-37°C, 120-180rpm shak...

Embodiment 1

[0071] A screening method for highly efficient oxytetracycline-degrading bacteria, comprising the following steps:

[0072] (1) Single colony culture:

[0073] The pig manure contaminated by oxytetracycline was taken to prepare a bacterial suspension, and the bacterial suspension was added to the basal medium. After the culture medium was solidified, the culture dish was inverted and cultured in a constant temperature incubator at 35°C for 3 days. After that, typical colonies with different shapes and clear colonies were selected for culture, and a single bacterial colony was cultivated.

[0074] The basal medium is prepared as follows: accurately weigh 10.0 g of peptone, 3.0 g of beef extract, 5.0 g of sodium chloride, and 15 g of agar, heat and dissolve in 1 L of distilled water, adjust the pH to 7.0, and sterilize at high temperature and high pressure for 15 minutes ; After the culture medium was cooled to 48° C., filter-sterilized cycloheximide was added so that the conce...

Embodiment 2

[0082] A screening method for highly efficient oxytetracycline-degrading bacteria, comprising the following steps:

[0083] (1) Single colony culture:

[0084] The pig manure contaminated by oxytetracycline was taken to prepare a bacterial suspension, and the bacterial suspension was added to the basal medium. After the culture medium was solidified, the culture dish was inverted and cultured in a constant temperature incubator at 35°C for 3 days. After that, typical colonies with different shapes and clear colonies were selected for culture, and a single bacterial colony was cultivated.

[0085]The basal medium is prepared as follows: accurately weigh 12.0 g of peptone, 5.0 g of beef extract, 6.0 g of sodium chloride, and 20 g of agar, heat and dissolve them in 1 L of distilled water, adjust the pH to 7.4, and sterilize at high temperature and high pressure for 30 minutes ; After the culture medium was cooled to 50° C., filter-sterilized cycloheximide was added so that the c...

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Abstract

The invention discloses a bacillus cereus as well as, a screening method, a preparation and application thereof. The bacillus cereus has efficient oxytetracycline degrading capability, and is screened according to the following method: (1) selecting pig manure bacterial suspension contaminated by oxytetracycline, and culturing single bacterial colonies in a culture medium containing 400-500mug / ml of cycloheximide; (2) continually improving the concentration of the oxytetracycline in the culture medium, to retain adaptive single bacterial colonies; and (3) culturing the single bacterial colonies in an inorganic salt culture medium, continually improving the concentration of the oxytetracycline, and preserving bacteria of which the oxytetracycline degrading rate is not less than 50% and the OD600 is not less than 0.2. The preparation comprises dried bacillus cereus powder and organic matrix in a mass ratio of 1:4 to 1:5. The bacillus cereus is applied to pig manure composting, has strong adaptability, high oxytetracycline degrading rate, low environmental influence and low cost, and is suitable for large-scale application.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and more specifically relates to a bacillus cereus, its screening method, preparation and application. Background technique [0002] Antibiotics are widely used in animal husbandry in my country, and are discharged into the environment with animal feces and urine in their bulk or transformed form, and accumulate in the environment, causing antibiotic pollution in the environment. When antibiotics enter the environment, they will destroy the balance of the original microbial species in the environment, which will lead to the imbalance of the corresponding ecosystem. In addition, oxytetracycline in the environment can also enter the human body through the biological chain, posing a threat to human health. To this end, the Ministry of Science and Technology of my country launched the National High-Tech Research and Development Program (863 Program), and set up a major special project of "Int...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F17/00C05F3/00C12R1/085
CPCC05F3/00C05F17/00C12N1/20C12N1/205C12R2001/085C05F11/08Y02W30/40Y02A40/20
Inventor 陈朱蕾汪佳席爽宫千惠俞瑛健
Owner HUAZHONG UNIV OF SCI & TECH
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