Application of indol or indol and calcium-magnesium ions in promoting formation of marine bdellovibrio sp. leech plastid

A technology based on calcium and magnesium ions and Bdellovibrio, which is applied in the direction of bacteria, microorganism-based methods, and the addition of compounds to stimulate growth, etc., can solve the problems of low survival rate of live bacteria and short shelf life, and achieve broad cracking spectrum properties and shelf life. Excellent prolongation and cracking effect

Active Publication Date: 2018-04-20
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
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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 swi

Method used

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  • Application of indol or indol and calcium-magnesium ions in promoting formation of marine bdellovibrio sp. leech plastid
  • Application of indol or indol and calcium-magnesium ions in promoting formation of marine bdellovibrio sp. leech plastid
  • Application of indol or indol and calcium-magnesium ions in promoting formation of marine bdellovibrio sp. leech plastid

Examples

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Effect test

Embodiment 1

[0052] The isolation and purification of embodiment 1 Bdellovibrio BDN-1

[0053] 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 ...

Embodiment 2

[0067] The promoting effect of embodiment 2 indole to Bdellovibrio BDN-1 leech plastid

[0068] (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;

[0069] (2) BDN-1 seed liquid culture: Pick several round plaques on the isolated plate, put them into 50mL DNB liquid medium, and then add 25...

Embodiment 3

[0074] Embodiment 3 Calcium ion promotes the culture effect to Bdellovibrio BDN-1 leech plastid

[0075] (1) Inoculate the host and cultivate: the same as step (1) in Example 2;

[0076] (2) BDN-1 seed liquid culture: with embodiment 2 step (2);

[0077] (3) calcium ion group inoculation and cultivation: with embodiment 2 step (3), this variable of indole is changed into the calcium chloride that final concentration is 4mM, 20mM and 40mM;

[0078] (4) Separation of leech plastids and double-layer inversion: with embodiment 2 step (4);

[0079] (5) result observation: with embodiment 2 step (5) (see Figure 4 ).

[0080] 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 ion concentration, the density of Bdellovibrio bdruplasma decreased.

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Abstract

The invention discloses application of indol or indol and calcium-magnesium ions to promotion of the formation of marine bdellovibrio sp. leech plastid, belonging to the technical field of bdellovibrio sp.. In the application, the indol or the indol and calcium-magnesium ions are utilized to promote the formation of bdellovibrio sp. BND-1 leech plastid; The indol or the indol and calcium-magnesiumions are determined to have an obvious effect with respect to prolonging the expiration date of a bdellovibrio sp. microbial preparation; and the indol or the indol and calcium-magnesium ions can beused for maintaining the vitality of thalluses in the bdellovibrio sp. microbial preparation. The high-density leech plastid provided by the invention can be used for inhibiting vibrioes capable of triggering biological diseases in aquaculture and has broad lysis spectrum performance, and the splitting action of the leech plastid is more excellent compared with that of a bdellovibrio sp. BDN-1 mixture (containing both the leech plastid and swinging bodies). The cultivating method for promoting the leech plastid is simple and feasible, suitable for promotion and application, and is expected tomake a contribution to preventing diseases in the aquaculture industry.

Description

technical field [0001] The invention belongs to the technical field of Bdellovibrio, and in particular relates to an application of indole or indole and calcium and magnesium ions in promoting the formation of marine Bdellovibrio baludoplasm. 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 to the aquaculture industry every year. [0003] At present, in aquaculture, antibiotics and chemical drugs are sti...

Claims

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Application Information

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IPC IPC(8): C12N1/38C12N1/20C12R1/01
CPCC12N1/20C12N1/38
Inventor 蔡俊鹏陈丹
Owner SOUTH CHINA UNIV OF TECH
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