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Boron-resistant lysinibacillus sp. with bacteriostatic ability and application of boron-resistant lysinibacillus sp.

A technology of Bacillus lysine and ability, applied in the field of microorganisms, can solve the problems of poor clinical use effect, undeveloped and used, etc., and achieve the effects of low production cost, high safety and good application prospects.

Active Publication Date: 2021-09-21
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, Bacillus subtilis, Bacillus coagulans, and Bacillus licheniformis are commonly used in livestock and poultry breeding, but with the extensive use of these strains, the clinical application effect is not good, and it is urgent to discover and identify new strains to update and iterate to meet different clinical needs need
[0004] Boron-resistant lysine bacillus is an important type of bacillus, which is widely used in the field of environmental management. It can degrade the concentration of arsenic in the soil, promote calcium carbonate deposition by changing the structure of branched chain fatty acids in the cell wall, and quickly Harmless treatment of livestock and poultry manure, but has not been developed and used in livestock and poultry breeding and disease prevention and control

Method used

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  • Boron-resistant lysinibacillus sp. with bacteriostatic ability and application of boron-resistant lysinibacillus sp.
  • Boron-resistant lysinibacillus sp. with bacteriostatic ability and application of boron-resistant lysinibacillus sp.
  • Boron-resistant lysinibacillus sp. with bacteriostatic ability and application of boron-resistant lysinibacillus sp.

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Embodiment 1

[0031] The isolation of embodiment 1 boron-resistant lysine bacillus LBS-2018

[0032] Green onions selected from the forage breeding experiment field of Jiangsu Academy of Agricultural Sciences were used as screening samples, and Escherichia coli ATCC25922 and clinically isolated multidrug-resistant Escherichia coli 3001 were used as indicator bacteria to carry out the test.

[0033] Take 1-2g green onion sample, grind and pulverize, add 1mL sterile PBS, treat in water bath at 80°C for 20min, inoculate LB medium at 2% ratio, and incubate at 37°C, 200r / min for 12h. For gradient dilution, draw 100 μL of the suspension for each gradient and evenly spread it on a sterile solid LB plate, and culture overnight (generally more than 12 hours) in a 37°C incubator. After colonies grow out, bacteria are randomly selected for purification.

[0034] Using double-layer agar point inoculation method, the concentration of indicator bacteria was adjusted to 1×10 6 CFU / mL, pipette and add 10...

Embodiment 2

[0035] The identification of embodiment 2 boron-resistant lysine bacillus LBS-2018

[0036] 1. Observation of strain morphology Inoculate LBS-2018 three-section line on LB plate, culture at 37°C, and observe the colony morphology the next day, as shown in figure 1 ; Use an inoculation loop to pick an appropriate amount of bacterial lawn, spread it on a clean glass slide with a drop of distilled water, air-dry and fix it, then use crystal violet mixed solution, iodine solution, 95% ethanol and safranin solution to stain in turn, wash and air-dry, and mirror check, such as figure 2 .

[0037] 2. 16S rDNA sequence analysis Kit was used to extract the DNA of LBS-2018, using primers 27F: 5′-AGAGTTTGATCCTGGCTCAG-3′ (SEQ ID NO.2), 1492R: 5′-TACGGTACCTTGTTACGACTT-3′ (SEQ ID NO.3) Carry out PCR amplification, and the composition of the PCR reaction system: 12.5 μL of 2×Taq PCR Master Mix, 1 μL of primers 27F and 1492R, 1 μL of template DNA, and finally make up to 25 μL with steriliz...

Embodiment 3

[0049] Example 3 Preparation of LBS-2018 bacterial agent

[0050] Take LBS-2018 out of the low-temperature storage refrigerator, scrape an appropriate amount of culture from the inoculation loop, inoculate the LB agar plate in three sections, and culture it upside down in a 37°C constant temperature incubator for 12h-20h. Take it out the next day, pick up a single colony and transfer it to LB culture medium, culture it at 37°C for 12-20 hours, and use it as the activated LBS-2018 strain. The activated strains were inoculated into new LB culture medium at a ratio of 1-2%, and cultured at the optimal culture temperature of 32°C and 200rpm for 20-24h with shaking for subsequent experiments.

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Abstract

The invention discloses a boron-resistant lysinibacillus sp. with bacteriostatic ability and application of the boron-resistant lysinibacillus sp.. The invention relates to a boron-resistant lysinibacillus sp. LBS-2018 with good bacteriostatic ability , which is preserved in the China Center for Type Culture Collection on December 17, 2020, and the preservation number is CGMCC No.21392. The invention also discloses an application of the boron-resistant lysinibacillus sp. LBS-2018 in preparation of a preparation for inhibiting escherichia coli, preferably an application of the boron-resistant lysinibacillus sp. LBS-2018 in preparation of a preparation for inhibiting drug-resistant escherichia coli. The LBS-2018 has a remarkable antibacterial effect on gram-negative bacteria (typically escherichia coli), particularly drug-resistant strains, the growth speed is higher than that of bacillus subtilis, and the drug-resistant bacteria are inhibited in a competitive mode. Compared with other boron-resistant lysinibacillus sp. strains, the boron-resistant lysinibacillus sp. strain disclosed in the invention has a stronger effect of inhibiting escherichia coli adhesion cells, and the inhibition rate is 61-81%.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a strain of boron-lysine-resistant bacillus with antibacterial ability and application thereof. Background technique [0002] Antibiotic resistance has become a major threat to global public health, leading to the rapid emergence and widespread spread of multidrug-resistant bacteria, posing a huge threat to human health and animal disease control. In order to ensure the sustainable development of human health and animal husbandry, the discovery and development of new antibiotics or antibiotic substitute products is imminent. The research and development of new antibiotics has a large investment and a long cycle, which is far behind the growth rate of drug-resistant bacteria, and the research and development of antibiotic substitutes has become a rigid demand. [0003] Bacillus is a Gram-positive bacterium that can survive under both aerobic and anaerobic conditions. It ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/742A61P31/04C12R1/07
CPCC12N1/20A61K35/742A61P31/04Y02A50/30
Inventor 钟小仙夏学峰钱晨张雪寒吴娟子刘智微
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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