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B.subtilis spores strain and application thereof

A technology of Bacillus subtilis and its application method, which is applied to Bacillus subtilis strains and its application fields, can solve problems affecting the microecological balance of livestock and poultry intestines, increase of drug-resistant pathogenic bacteria, and excessive drug residues in livestock and poultry products, and achieve improvement. Production performance, high enzyme production capacity, and broad application prospects

Active Publication Date: 2013-08-28
山东蔚蓝生物科技有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the problems caused by the abuse of antibiotics have attracted increasing attention, such as the increase of drug-resistant pathogens, excessive drug residues in livestock and poultry products, affecting the micro-ecological balance in the intestinal tract of livestock and poultry and the safety of livestock and poultry food, etc.

Method used

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  • B.subtilis spores strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Example 1 Isolation, screening and identification of Bacillus subtilis BW2:

[0011] 1. Primary screening of strains

[0012] Collect 6 fresh feces samples from healthy piglets, weigh 5g samples respectively, add 45mL sterile water to prepare a suspension, treat non-spore vegetative bodies in an 80°C water bath, spread on LB plates, and incubate at 37°C for 48 Hours, 48 ​​strains of bacillus were obtained.

[0013] 2. Screening of Enzyme-producing Bacillus

[0014] 1) α-amylase detection:

[0015] Starch medium: peptone 10g, beef extract 5g, NaCl 5g, soluble starch 2g, distilled water 1000ml, agar powder 20g.

[0016] Spot the isolated strains on the medium, culture at 37°C for 24-48 hours, drop an appropriate amount of iodine solution on the surface of the medium to evenly cover the entire plate, if a colorless transparent circle appears around the colony, it means that the starch has been hydrolyzed .

[0017] 2) Protease detection:

[0018] Milk medium: skim mi...

Embodiment 2

[0025] Embodiment 2: Bacillus subtilis BW2 enzyme production experiment

[0026] 1. α-amylase detection:

[0027] Starch medium: peptone 10g, beef extract 5g, NaCl 5g, soluble starch 2g, distilled water 1000ml, agar powder 20g.

[0028] Spot the isolated strains on the medium, culture at 37°C for 24-48 hours, drop an appropriate amount of iodine solution on the surface of the medium to evenly cover the entire plate, if a colorless transparent circle appears around the colony, it means that the starch has been hydrolyzed . Measure the ratio of the diameter of the transparent circle to the diameter of the colony.

[0029] 2. Protease detection:

[0030] Milk medium: skim milk 50ml, distilled water 50ml, agar powder 1.5g.

[0031] Inoculate the isolated strains into milk medium and incubate at 37°C for 24-48 hours. If there is a transparent circle around the colony, it means that the casein has been hydrolyzed. Measure the ratio of the diameter of the transparent circle to t...

Embodiment 3

[0037] Embodiment 3: the preparation of bacillus subtilis BW2 bacterium powder

[0038] 1. Seed solution preparation: Pick a ring of fresh Bacillus subtilis BW2 sludge, inoculate it in 100mL of LB medium (100mL of 500mL triangular bottle solution), incubate at 37°C and 200rpm for 12-16h to prepare the seed solution;

[0039] 2. Fermentation culture: Inoculate the above-mentioned seed solution with 5-10% inoculation amount into the sterilized nutrient broth medium, culture at 37°C and 200 rpm for 28-32 hours, and stop the fermentation culture when the yield of spores under the microscope is above 95%;

[0040] 3. Bacteria powder preparation: Centrifuge the fermentation broth (8000rpm, 5min) to collect the bacteria, add dextrin to stir and mix, vacuum freeze-dry, and make Bacillus subtilis BW2 bacteria powder with a bacterial content of 50-100 billion / g.

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Abstract

The invention aims at providing b.subtilis spores and application of the b.subtilis spores to livestock and poultry breeding. The b.subtilis spores is screened from a fresh fecal sample of a healthy piglet, has strong capacity in producing amylase, protease and cellulase, can be applied to a livestock feed as a feed additive, can effectively improve the production performance of breading an animal, and is wide in application prospect.

Description

technical field [0001] The invention belongs to the technical field of microbial screening and application, and in particular relates to a bacillus subtilis strain and its application. Background technique [0002] In livestock and poultry breeding, antibiotics are widely used in livestock and poultry feed because they can effectively reduce the incidence of livestock and poultry and improve the growth efficiency of livestock and poultry. However, the problems caused by the abuse of antibiotics have attracted increasing attention, such as the increase of drug-resistant pathogens, excessive drug residues in livestock and poultry products, affecting the micro-ecological balance in the intestinal tract of livestock and poultry, and the safety of livestock and poultry food. In this regard, the EU has completely banned the use of antibiotic growth-promoting feed additives in feed. In order to deal with these problems, the use of probiotic microecological preparations to replace ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16C12R1/125
Inventor 陈琳王增元
Owner 山东蔚蓝生物科技有限公司
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