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Bacillus velezensis as well as product and application thereof

A technology of bacillus and bacteria agent, applied in Bacillus velei and its products and application fields, can solve the problems of lack of biological control antagonistic bacteria, no application of prevention and control of aeromonas disease, etc., and achieve good application prospects, Good biosafety, effect of improving anti-infection ability

Inactive Publication Date: 2021-03-19
SHENZHEN INST OF GUANGDONG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology improves upon existing methods such as chemical or physical agents like formaldehydes (formaldehyde), antibiotics, enzyme catalysts, etc., but they are still being tested on animals due their negative sideings. By introducing Bacteria called Lysinsulphidium sppiesum into this process, researchers were able to demonstrate its potential use in controlling harmful microorganisms causing diseases caused by these organism's activity. These techniques could potentially lead to new ways to fight infectious airborne illnesses through immunization strategies targeted specifically specific strains within them.

Problems solved by technology

This patented technical problem addressed in this patents relates to controlling airborne infectious organism populations like Amanicola spirochetes through biocontrol agents targeting specific viruses called Aviridae while avoiding their resistance towards known pescides used worldwide due to concerns over safety issues associated with these substances.

Method used

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  • Bacillus velezensis as well as product and application thereof
  • Bacillus velezensis as well as product and application thereof
  • Bacillus velezensis as well as product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 The isolation of cultureable bacteria from Shenzhen mangrove deposits and the screening of antagonistic bacteria

[0053] (1) Isolation and culture of culturable bacteria from mangrove sediments in Shenzhen

[0054] On September 13, 2019, the sediment (5-10cm) was collected in the mangrove forest in Dongchong, Shenzhen, put into an incubator with ice packs and brought back to the laboratory for processing within 12 hours. Take 3g of sediment and add sterile seawater to dilute to 30mL, dilute the sample 10 times as the mother liquor, and then dilute the mother liquor 10 times to form 10 0 , 10 -1 , 10 -2 , 10 -3 , 10 -4 For a series of concentrations, take 100 μL of each concentration on 2216E solid medium (formula: 5g peptone, 1g yeast extract, 0.01g ferric phosphate, 1g beef extract, 1L seawater, pH7.4-7.6) and aseptically spread evenly. Each concentration was repeated 3 times. The coated medium plate was cultured upside down in a 28°C incubator for 24-...

Embodiment 2

[0059] Embodiment 2 Identification and growth characteristic analysis of antagonistic bacteria

[0060] (1) Identification of antagonistic bacteria

[0061] 16S rRNA sequence cloning and phylogenetic analysis: Genomic DNA of strain AQ1 was extracted using Ruizhen Biological Bacterial Genomic DNA Extraction Kit, and DNA was used as a template to amplify the target gene with 16S rRNA gene primers to obtain amplified products. The primer sequences are as follows:

[0062] Forward primer 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID No.1);

[0063] Reverse primer 1492R: 5'-GGTTACCTTGTTACGACTT-3' (SEQ ID No. 2).

[0064] The amplified product was observed and photographed by electrophoresis and gel imaging system, showing a single target band of about 1.5kb. Send the PCR product to Sangon Bioengineering Co., Ltd. for sequencing. The nucleotide sequence information is shown in SEQ ID No.3:

[0065]CTCTGTCACCTTCGGCGGCTGGCTCCTAAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTAC...

Embodiment 3

[0072] The antagonistic substance of embodiment 3 antagonistic bacteria first mentions its antibacterial effect evaluation

[0073] (1) Preliminary extraction of antagonistic substances

[0074] The antagonistic substance of AQ1 was initially extracted by concentrated hydrochloric acid precipitation method: the concentration was 1×10 8 The CFU / mL antagonistic bacteria AQ1 bacterial liquid was transferred to 200mL 2216E liquid medium at a ratio of 1:100 (V / V), and cultured on a constant temperature shaker at 28°C and 200r / min for 48-72h to obtain a fermentation broth. Take 50mL Bacteria solution was centrifuged at 8000rpm for 20min, and the supernatant was obtained after twice centrifugation. Slowly add 1 mol / L HCl solution to the supernatant until white flocs are precipitated, and the pH is about 2 at this time, and place at 4°C overnight. Centrifuge at 8000rpm for 20min, collect the precipitate and redissolve it with 3-5mL PBS (phosphate buffered saline), and filter through...

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Abstract

The invention provides bacillus velezensis as well as a product and application thereof, and relates to the technical field of biology. The bacillus velezensis is derived from mangrove forest sediments, and has obvious inhibiting effects on pathogenic bacteria including plesiomonas shigelloides, Edwardsiella, aeromonas hydrophila, aeromonas caviae, aeromonas veronii, aeromonas media, aeromonas schubertii and aeromonas salmonicida; and a fermentation liquid of the bacillus velezensis is used as a bait additive for animal experiments, it is found that the infection resistance of silver carp to the aeromonas can be remarkably improved, meanwhile, good biological safety is achieved, and therefore the bacillus velezensis has good application prospects in prevention and treatment of aeromonas diseases. The invention provides a biological preparation for preventing and treating aeromonas diseases. The preparation can be used for preventing and treating the aeromonas diseases.

Description

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Claims

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Application Information

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Owner SHENZHEN INST OF GUANGDONG OCEAN UNIV
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