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A kind of Bacillus subtilis with antibacterial and virus hemagglutination functions and application thereof

A Bacillus subtilis, functional technology, applied to Bacillus subtilis and its application fields, can solve the problems of the destruction of the intestinal microecological balance of poultry, the decrease of the immune function of poultry, the imbalance of intestinal flora, etc., and the antiviral effect is obvious and reduced. The effect of improving blood coagulation value and vomiting symptoms

Active Publication Date: 2022-04-26
山东蔚蓝生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because poultry growth, especially early growth, is easily affected by factors such as environment and nutritional stress, the intestinal microecological balance of poultry is easily damaged, resulting in a decrease in the immune function of poultry, causing an imbalance in the intestinal flora and reducing its production performance, causing various disease, even death

Method used

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  • A kind of Bacillus subtilis with antibacterial and virus hemagglutination functions and application thereof
  • A kind of Bacillus subtilis with antibacterial and virus hemagglutination functions and application thereof
  • A kind of Bacillus subtilis with antibacterial and virus hemagglutination functions and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Strain source and identification

[0029] 1. Sample source: duck intestinal mucosa of Pingdu Farm in Qingdao City, Shandong Province.

[0030] 2. Strain isolation and screening:

[0031] Scrape the duck intestinal mucosa with a glass slide, place it in a sterile centrifuge tube, add sterilized saline, get the bacterial suspension, place the bacterial suspension in an 80 °C water bath pot, treat it for 10 min, remove it, and cool it rapidly; dilute the treated bacteria suspension to a suitable concentration in a 10-fold gradient, take 100uL on the nutrient agar plate, apply evenly, culture at 37 °C for 16-24h; pick up the different single colonies that have been cultivated on the nutrient agar plate, further draw the line 3 times, and purify the single strain. The applicant selected five monobacteria and named them A, B, C, D, and E.

[0032] Inoculate 5 strains of bacteria into nutritious broth medium at 37 °C, 220 r / min for 14 h, centrifuge the bacterial solution...

Embodiment 2

[0040] Example 2 Identification of DB2572 strains

[0041] 2.1 Colony morphological identification

[0042] as Figure 1 As shown, the DB2572 strain is slightly yellowish on the nutrient agar medium, with a smooth and transparent surface and mucus.

[0043] 2.2 16S rRNA Molecular Identification

[0044] The genome of strain DB2572 was extracted using a kit. The genome is then used as a template to amplify its 16S rRNA using specific primers. PCR systems include: 0.7 μl 27F, 0.7 μl 1492R, 4 μl template DNA, 17.5 μl SuperMiX, and 12.1 μl water. PCR reaction conditions are set to: (1) 94 °C 5 min, (2) 94 °C prenaturation 30s, (3) 55 °C 30s; (4) 72 °C 1 min; perform steps (2) to (4) 35 cycles; (5) 72 °C 10 min. The PCR product was detected by 1% agarose gel electrophoresis and sequenced to obtain the strain 16S rRNA sequence SEQ ID NO:1.

[0045] BY COMPARING SEQ ID NO:1 with BLAST in the NCBI database, it was found that it had the highest similarity with Bacillus subtilis. Therefore,...

Embodiment 3

[0061] Example 3 Bacillus subtilis DB2572 evaluation of the ability to reduce viral blood coagulation

[0062] 1. Preparation of centrifugal solution for test bacteria

[0063] Bacillus subtilis DB2572 1 inoculation loop was inoculated into LB medium at 37 °C, 220 r / min for 48 h, and the spore yield was above 90% by microscopic examination; the bacterial solution was centrifuged at 10000 r / min for 5 min, the bacterial solution was supernatanted, and the 0.22 μm filter was used for later.

[0064]2. Preparation of dilutions of avian influenza virus and Newcastle disease virus

[0065] H9N2 avian influenza seed poison, VGGA newcastle disease seed poison, with sterile PBS to make 5000 times dilution for backup.

[0066] 3. Evaluation of the effect of Bacillus subtilis DB2572 in reducing the HA of avian influenza virus and Newcastle disease virus

[0067] After the diluted avian influenza seed poison and Newcastle disease seed poison were mixed with the supernatant volume of Bacillus...

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Abstract

The invention relates to the technical field of functional microorganism screening and application, and provides a bacillus subtilis with functions of inhibiting bacteria and reducing virus hemagglutination and its application. The Bacillus subtilis has been deposited in the China Center for Type Culture Collection of Wuhan University, China on November 16, 2020, and the deposit number is CCTCC NO: M2020744. The bacillus subtilis can effectively inhibit various pathogenic bacteria, and at the same time can significantly reduce the hemagglutination value of viruses such as avian influenza and Newcastle disease, and can be widely used as a feed additive for the prevention and control of common diseases of livestock and poultry.

Description

Technical field [0001] The present invention relates to functional microbial screening and application technology, specifically to a bacillus subtilis having a bacteriostatic and reducing viral hemagglutination function and its applications. Background [0002] The irrational application of antibiotics in the aquaculture industry not only endangers animal health, but also brings many drawbacks, such as drug resistance, drug residues and food safety. Probiotics are a class of microorganisms with biological activity and beneficial to host health, and because of their non-polluting, non-toxic and non-adverse reactions, antibacterial health care and growth promotion, etc., they have become a new type of additive after antibiotics. In poultry production, the application of probiotics can play a variety of roles in the health of poultry intestinal microecology, the most direct of which is to optimize the intestinal flora and improve the intestinal microecological environment. Because ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/742A61P31/16A61P31/14A61P1/12A23K10/18A23K50/75C12R1/125
CPCA61K35/742A61P31/16A61P31/14A61P1/12A23K10/18A23K50/75
Inventor 李宁凌红丽彭虹旎周英俊陈琳车传燕郭宝帅
Owner 山东蔚蓝生物科技有限公司
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