Application of Indole in Promoting the Formation of Marine Bdellovibrio Bacteroids
A technology of Bdellovibrio and indole, 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 affecting product market development, low survival rate of live bacteria, low survival rate, etc., and achieve the goal of cultivating The method is simple and easy to implement, has good antibacterial effect and the effect of inhibiting Vibrio
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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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