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Application of indole in promoting formation of marine Bdellovibrio sp. BDE-1 bdelloplast

A leech vibrio, indole technology, applied in bacteria, microorganism-based methods, and adding compounds to stimulate growth, etc., can solve the problems affecting product market development, low survival rate of viable bacteria, short shelf life, etc., and achieve the cultivation method. Simple and easy to operate, good antibacterial effect, and the effect of prolonging the shelf life

Active Publication Date: 2018-06-01
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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  • Application of indole in promoting formation of marine Bdellovibrio sp. BDE-1 bdelloplast
  • Application of indole in promoting formation of marine Bdellovibrio sp. BDE-1 bdelloplast
  • Application of indole in promoting formation of marine Bdellovibrio sp. BDE-1 bdelloplast

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0048] Retrieve samples from an aquaculture seawater area in Hainan, use Bacillus subtilis as the parasitic host bacteria, take 10mL of sample liquid and add it to aseptically treated 50mL DNB liquid medium, enrich culture at 200r / min, 30°C After 30 hours, centrifuge the bacterial 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 to 4 days, select a plate with 30 to 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 double-layer solid med...

Embodiment 2

[0063] The promoting effect of embodiment 2 indole to Bdellovibrio BDE-1 leech plastid

[0064](1) Inoculate the host and cultivate: take a bottle of newly cultivated host seed solution of Bacillus subtilis GIM1.136, burn the mouth of the bottle under the outer flame of the alcohol lamp in the aseptic operation table, and absorb 3mL of the host fungus seed Transfer the solution into 250mL bottled NB liquid medium, place the inoculated host bacteria in a constant temperature shaker at about 31°C for about 15 hours, then divide into four 100mL centrifuge tubes, and put them into a refrigerated centrifuge Centrifuge at 6000r / min at 4°C for 10min, then discard the supernatant, add 1mL of sterile distilled water with a salinity of 5‰ to each tube to suspend, mix well, transfer to a centrifuge tube, and cover with a sterile cap. Bagging, the bag needs to be sterile, and then put it in a refrigerator at 4°C for future use to obtain the host bacteria concentrate;

[0065] (2) BDE-1 s...

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 application of indole in promoting formation of a marine Bdellovibrio sp. BDE-1 bdelloplast, and belongs to the technical field of Bdellovibrio sp. BDE-1. According to the application of the indole in promoting the formation of the marine Bdellovibrio sp. BDE-1 bdelloplast, a factor, namely, the indole, is formed by utilizing a Bdellovibrio sp. BDE-1 promoting bdelloplast,so that the formation of high-density bdelloplast of the Bdellovibrio sp. BDE-1 is promoted; the research discovers that the indole has an obvious promoting effect on the formation of the bdelloplastof the Bdellovibrio sp. BDE-1; the cracking performance of the bdelloplast is not reduced; compared with a control group, the quality guarantee period of a high-density bdelloplast group is obviouslyprolonged; the high-density bdelloplast provided by the invention can inhibit vibrios which can cause human infection and food poisoning and can also inhibit the vibrios which can cause biological diseases in aquaculture, has a very good inhibiting effect on other food-borne pathogenic bacteria, and has wide cracking spectrum; the cracking effect of the bdelloplast is unchanged as compared with that of a BDE-1 mixture; and a bdelloplast promoting culturing method disclosed by the invention is simple and feasible, 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 indole in promoting the formation of marine Bdellovibrio bloclasts. 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 gastroenteritis in huma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/38C12N1/20C12N1/04C12R1/01
CPCC12N1/04C12N1/20C12N1/38
Inventor 蔡俊鹏李敏佳
Owner SOUTH CHINA UNIV OF TECH
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