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The Application of Calcium Ion in Promoting the Formation of Bdellovibrio Bdellovibrio Plastids

A technology of Bdellovibrio and calcium ions, which is applied in the direction of application, bacteria, and fungicides, can solve the problems of affecting product market development, low survival rate of viable bacteria, and low survival rate, and achieves simple and easy cultivation methods and broad cracking Spectrum, the effect of inhibiting Vibrio

Active Publication Date: 2020-09-22
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Our research also shows that at present, Bdellovibrio has the problems of extremely low survival rate and extremely short shelf life, which affects the market development of products
[0008] It can be seen that it has important economic value to solve the problems of low survival rate of live bacteria and short shelf life in current Bdellovibrio products

Method used

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  • The Application of Calcium Ion in Promoting the Formation of Bdellovibrio Bdellovibrio Plastids
  • The Application of Calcium Ion in Promoting the Formation of Bdellovibrio Bdellovibrio Plastids
  • The Application of Calcium Ion in Promoting the Formation of Bdellovibrio Bdellovibrio Plastids

Examples

Experimental program
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Effect test

Embodiment 1

[0047] The isolation and purification of embodiment 1 Bdellovibrio BDE-1

[0048] The samples were retrieved from a certain aquaculture seawater area in Hainan, Bacillus subtilis was used as the parasitic host bacteria, 10 mL of sample liquid was taken and added to 50 mL of aseptically treated DNB liquid, enriched at 200 r / min at 30 ° C for 30 h, and the bacteria Centrifuge the solution at 6000r / min for 10min, then centrifuge the supernatant at 20000r / min for 20min, suspend a small amount of bacteria at the bottom of the centrifuge tube with sterile water, and dilute the suspension to 10 -1 、10 -2 、10 -3 、10 -4 , pour each dilution gradient on a DNB double-layer solid medium plate in turn, culture at 30°C for 3-4 days, select a plate with 30-300 plaques, and pick the plate with the same size, round, transparent and neat edges with the inoculation needle The single phage plaques were cultured in 50mL DNB liquid medium, and then continued to centrifuge and dilute the DNB doub...

Embodiment 2

[0063] Embodiment 2 Calcium ions promote the culture action of Bdellovibrio BDE-1 leech plastid

[0064] (1) Inoculate the host and cultivate: Take a bottle of the latest cultured host Bacillus subtilis (Bacillus subtilisGIM 1.136) host seed solution, burn the mouth of the bottle under the outer flame of the alcohol lamp in the aseptic operation table, draw 3mL of the host bacteria seed solution and transfer to Put it into 250mL bottled NB liquid medium, culture it in a constant temperature shaker at about 31°C for about 15h, then divide it into four 100mL centrifuge tubes, put it into a refrigerated centrifuge at 6000r / min, 4°C Centrifuge for 10 minutes, then discard the supernatant, add 1mL sterile distilled water with a salinity of 5‰ to each tube to suspend, disperse and mix evenly, transfer it into a centrifuge tube, cover with a sterile cover, and bag the bag. The bag needs to be sterile, and then Store in a 4°C refrigerator for later use to obtain a concentrated solution ...

Embodiment 3

[0074] The application of the wide cleavage spectrum of embodiment 3 Bdellovibrio BDE-1 leech plastids

[0075] (1) Preparation of 39 strains of indicator bacteria

[0076] Inoculate a single colony of 39 strains of indicator bacteria (Table 2) into the nutrient broth liquid medium, culture at 37°C and 200r / min for 8h, and adjust its concentration to 1×10 6 CFU / mL, stored at 4°C, for later use;

[0077] Table 2 39 different experimental indicator strains

[0078]

[0079]

[0080] (2) Preparation of Bdellovibrio Bdeloidoplasts

[0081] Bdellovibrio sp. (Bdellovibrio sp.) BDE-1 single phage plaque identified in Example 1 was inoculated in 50mL DNB liquid medium, 30 ℃, 2000r / min, cultivated for 3 days, cultivated its concentration to 1 × 10 8 PFU / mL; 6000r / min, 4 ℃, 10min centrifugation Bdellovibrio culture fluid, the enrichment precipitate (Belichoplasma) is standby for bacterial lysing experiment, 4 ℃ preserves, standby (the Bdellovibrio concentration of the mixed sol...

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Abstract

The invention discloses an application of calcium ions in promoting the formation of marine bdellovibrio bdelloplasts, and belongs to the technical field of bdellovibrio. The present invention promotes the formation of Bdellovrio sp. Bdellovrio sp. BDE‑1 high-density Bdellovrio sp. Bdellovrio sp. by utilizing calcium ions, the factor that promotes the formation of Bdellovibrio leech plastids; The formation of hiruplasma all has obvious promoting effect, and the cracking performance of hiruplasma has not declined, and the shelf life of high-density hiruplasma group has been obviously prolonged compared with control group; The Vibrio that causes human infection and food poisoning can also inhibit the Vibrio that can cause biological diseases in aquaculture. At the same time, it also has a good antibacterial effect on other food-borne pathogens. It has a wide cracking spectrum and its cracking Compared with the mixture of Bdellovibrio BDE-1, the effect remains unchanged; the method for promoting the cultivation of the bdeloid plastids of the present invention is simple and easy, and can be popularized and used.

Description

technical field [0001] The invention belongs to the technical field of Bdellovibrio, and in particular relates to an application of calcium ions in promoting the formation of marine Bdellovibrio bdelloplasts. Background technique [0002] Vibrio (Vibrio) is one of the most common bacterial groups in the marine environment, widely present in seawater, seabed sediments, and seafood such as fish, shrimp, shellfish, crabs, sea cucumbers, and kelp. The 8th edition of "Bergey's Handbook of Bacteriology" recorded 35 species of Vibrio, a considerable part of which are important pathogenic bacteria of aquaculture organisms. In the aquaculture industry, the diseases of cultured organisms caused by Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio anguillarum, etc. are collectively called vibriosis, which causes huge economic losses to the aquaculture industry every year. At the same time, some Vibrios are also pathogenic bacteria of humans. They can not only cause gastroenteriti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/38A01N63/20A01P3/00A01P1/00C12R1/01
CPCA01N63/00A01N63/10C12N1/20C12N1/38C12N1/205C12R2001/01
Inventor 蔡俊鹏李敏佳
Owner SOUTH CHINA UNIV OF TECH
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