A strain of marine Bdellovibrio and its application to promote the formation of hiruplasma under ampicillin

A technology of ampicillin and Bdellovibrio, which is applied in the fields of application, bacteria, and fungicides, can solve the problems of short shelf life and low survival rate of viable bacteria, and achieve the effects of extended shelf life, wide cleavage spectrum properties, and simple and feasible cultivation methods

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

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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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  • A strain of marine Bdellovibrio and its application to promote the formation of hiruplasma under ampicillin
  • A strain of marine Bdellovibrio and its application to promote the formation of hiruplasma under ampicillin
  • A strain of marine Bdellovibrio and its application to promote the formation of hiruplasma under ampicillin

Examples

Experimental program
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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 a strain of marine bdellovibrio and the application of promoting the formation of bdellovibrio under ampicillin, and belongs to the technical field of bdellovibrio. The strain is Bdellovibrio sp. BDN‑1, which was preserved on November 27, 2017 at the Guangdong Provincial Microbiological Culture Collection Center, 5th Floor, Building 59, Compound 59, No. 100 Xianlie Middle Road, Guangzhou City , deposit number: GDMCC NO: 60291. The present invention utilizes ampicillin or ampicillin and calcium and magnesium ions to promote the formation of the Bdellovibrio Bdellovibrio Bideloplasts; it has been confirmed that it has the effect of significantly prolonging the shelf life of Bdellovibrio microbial preparations effect; and can effectively maintain the vitality of the bacterium in the Bdellovibrio microbial preparation for a long time. The method for promoting leech plastid cultivation provided by the invention is simple and feasible, is suitable for popularization and use, and is expected to contribute to the prevention and control of diseases 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 Patents(China)
IPC IPC(8): C12N1/20C12N1/38A01N63/20A01P3/00A01P1/00C12R1/01
CPCA01N63/00A01N63/10C12N1/20C12N1/38C12N1/205C12R2001/01
Inventor 蔡俊鹏陈丹
Owner SOUTH CHINA UNIV OF TECH
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