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Marine bdellovibrio and application thereof in promoting formation of bdelloplast under existence of ampicillin

A technology of ampicillin and leech vibrio, applied in the application, bacteria, fungicides and other directions, can solve the problems of low survival rate of viable bacteria and short shelf life, and achieve the effect of wide cracking spectrum, extended shelf life and improved cracking ability.

Active Publication Date: 2018-05-01
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the key to solve the problems of low survival rate and short shelf life of the current Bdellovibrio products is to transform the swimming body into the bructoria during the high-concentration fermentation process, and at the same time delay the development of the bdeltoplast.

Method used

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  • Marine bdellovibrio and application thereof in promoting formation of bdelloplast under existence of ampicillin
  • Marine bdellovibrio and application thereof in promoting formation of bdelloplast under existence of ampicillin
  • Marine bdellovibrio and application thereof in promoting formation of bdelloplast under existence of ampicillin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Isolation and purification of Bdellovibrio BDN-1

[0057] Retrieve the sample from an aquaculture seawater area in Fujian, take 10mL of the sample liquid and centrifuge it at 20000rpm for 20min to enrich, then suspend it with sterilized DNB liquid medium, add it into the aseptically treated 50mL bottled DNB liquid nutrient base, Cultivate at 200r / min at 30°C for 48h, centrifuge the bacterial solution at 6000rpm for 10min, take the supernatant and filter it through a 0.8μm cellulose acetate filter membrane, centrifuge the filtrate at 20000rpm for 20min, suspend the sediment with sterile water, and diluted to 10 -1 、10 -2 、10 -3 、10 -4 , Pour each dilution gradient onto the DNB double-layer solid medium plate to detect plaques, culture at 30°C for 3 to 4 days, pick a single phage plaque with the same size, round, transparent and neat edges from the inoculation loop, and culture it in 50mL DNB liquid Continue to culture in the culture medium, pour the DNB dou...

Embodiment 2

[0071] Example 2 The effect of ampicillin on promoting the cultivation of Bdellovibrio BDN-1 leech plastids

[0072] (1) Host culture: Take the newly cultivated host bacteria Bacillus subtilis GIM1.136, burn the mouth of the bottle under the outer flame of the alcohol lamp in the aseptic operation table, absorb 3mL of the host bacteria seed liquid and transfer it into a 250mL bottle of NB liquid In the culture medium, put the inoculated host bacteria in a constant temperature shaker at about 31°C for about 15 hours, then put them in a refrigerated centrifuge at 6000rpm and centrifuge at 4°C for 10min, then discard the supernatant, add to each tube 1mL of sterile distilled water with a salinity of 15‰, dispersed and mixed evenly, transferred to a centrifuge tube to obtain the host bacteria concentrate, bagged, and stored in a 4°C refrigerator for later use;

[0073] (2) BDN-1 seed liquid culture: Pick several round plaques on the isolated plate, put them into 50mL DNB liquid medi...

Embodiment 3

[0078] Example 3 Calcium ions promote the cultivation of Bdellovibrio BDN-1 leech plastids

[0079] (1) Inoculate the host and cultivate: the same as step (1) in Example 2;

[0080] (2) BDN-1 seed liquid culture: with embodiment 2 step (2);

[0081] (3) Calcium ion group inoculation and cultivation: with embodiment 2 step (3), the calcium chloride that final concentration is 4mM, 20mM and 40mM is changed into factor variable;

[0082] (4) Separation of leech plastids and double-layer inversion: with embodiment 2 step (4);

[0083] (5) result observation: with embodiment 2 step (5) (see Figure 4 ).

[0084] Results Compared with the control group, the calcium ion concentrations of 4mM, 20mM and 40mM decreased by 91%, 93% and 99% respectively. With the increase of calcium ions, the density of Bdellovibrio bruloglast decreased.

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Abstract

The invention discloses marine bdellovibrio and application thereof in promoting the formation of bdelloplast under existence of ampicillin, and belongs to the technical field of bdellovibrio. The marine bdellovibrio is (Bdellovibrio sp.) BDN-1, and was preserved into Guangdong Microorganism Center for Culture Collection in Guandong Microorganism Research Institute (No.5 Floor, No.59 Building, No.100 Central Xian Lie Middle Road, Guangzhou City) on November 27th, 2017, and the preservation number is GDMCC NO:60291. The marine bdellovibrio has the advantages that the factors for promoting the formation of the bdelloplast by the bdellovibrio, such as ampicillin or ampicillin and calcium and magnesium ions, are utilized, and the formation of the bdelloplast by the bdellovibrio BND-1 is promoted; the effect of obviously extending the quality warranty period of the bdellovibrio microorganism preparation is verified; the activity of the bacteria in the bdellovibrio microorganism preparationis effectively maintained for a long time; the culture method for promoting the formation of the bdelloplast is simple and feasible, and is suitable for being popularized and applied, and the contribution is expected to be made to the disease and pest controlling in the aquaculture industry.

Description

technical field [0001] The invention belongs to the technical field of Bdellovibrio, and in particular relates to a marine Bdellovibrio with a wide lysis spectrum which takes Bacillus subtilis (Bacillus subtilis) as a parasitic host and promotes high-density hirudine under ampicillin or ampicillin and calcium and magnesium ions. body forming applications. Background technique [0002] Vibrio (vibrios) 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/38A01N63/02A01N63/00A01P3/00A01P1/00C12R1/01
CPCA01N63/00A01N63/10C12N1/20C12N1/38C12N1/205C12R2001/01
Inventor 蔡俊鹏陈丹
Owner SOUTH CHINA UNIV OF TECH
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