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Marine bdellovibrio sp. and application of marine bdellovibrio sp. to facilitation of formation of leech plastid in presence of ampicillin

A technology of ampicillin and Bdellovibrio, which is applied in the field of marine Bdellovibrio, can solve problems such as short shelf life, low survival rate, and impact on product market development, and achieve good antibacterial effect, extended shelf life, and simple and easy cultivation methods Effect

Active Publication Date: 2018-05-04
SOUTH CHINA UNIV OF TECH
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  • Description
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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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  • Marine bdellovibrio sp. and application of marine bdellovibrio sp. to facilitation of formation of leech plastid in presence of ampicillin
  • Marine bdellovibrio sp. and application of marine bdellovibrio sp. to facilitation of formation of leech plastid in presence of ampicillin
  • Marine bdellovibrio sp. and application of marine bdellovibrio sp. to facilitation of formation of leech plastid in presence of ampicillin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The isolation and purification of embodiment 1 Bdellovibrio BDE-1

[0052] 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 into a double-layer plate in turn, culture at 30°C for 3-4 days, select a plate with 30-300 plaques, and pick a single phage that is basically the same size, round, transparent and with neat edges with the inoculation needle Put the plaques into 50mL DNB liquid to continue culturing, then continue to centrifuge and dilute and pour the d...

Embodiment 2

[0067] Embodiment 2 ampicillin promotes the culture effect to Bdellovibrio BDE-1 leech plastid

[0068] (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 the inoculated host bacteria 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, put them in 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, disperse and mix evenly, transfer to a centrifuge tube, cover with a sterile cap, and bag , the bag needs to be aseptic, and then put it in a 4°C refrigerator for late...

Embodiment 3

[0078] The application of the wide cleavage spectrum of embodiment 3 Bdellovibrio BDE-1 leech plastids

[0079] (1) Preparation of 39 strains of indicator bacteria

[0080] 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;

[0081] Table 2 39 different experimental indicator strains

[0082]

[0083]

[0084] (2) Preparation of Bdellovibrio Bdeloidoplasts

[0085] 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 marine bdellovibrio sp. and application of the marine bdellovibrio sp. to facilitation of formation of leech plastid in the presence of ampicillin. The bacterial strain is bdellovibrio sp. (BED-1) which is collected in Guangdong Microbiological Culture Collection Center of Guangdong Institute of Microbiology which is located at 5th floor, building 59, No. 100 Courtyard, Xianlie Middle Road, Guangzhou on November 27, 2017 with the collection number GDMCC NO:60292. The formation of high-density leech plastid of the bdellovibrio sp. BDE-1 is facilitated with the ampicillinserving as a factor for facilitating the formation of the leech plastid of the bdellovibrio sp., so that high-concentration leech plastid of the bdellovibrio sp. BDE-1 is provided. As found by the research, the ampicillin plays a role in remarkably facilitating the formation of the leech plastid of the bdellovibrio sp., and the cracking performance of the leech plastid does not decrease; comparedwith a control group, the shelf life of the high-density leech plastid is prolonged remarkably; the method for culturing the leech plastid 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 a marine Bdellovibrio with a wide lysis spectrum which uses Bacillus subtilis as a parasitic host and an application of promoting the formation of high-density bructorids under ampicillin. 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...

Claims

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Application Information

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IPC IPC(8): C12N1/38C12N1/20A61K35/741A61P31/04C12R1/63
CPCA61K2035/115C12N1/20C12N1/38Y02A50/30
Inventor 蔡俊鹏曹青青李敏佳
Owner SOUTH CHINA UNIV OF TECH
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