A kind of Pseudomonas bacterial strain and its application
A technology of Pseudomonas and Aeromonas, applied in the field of microorganisms, can solve problems such as high incidence, economic loss in aquaculture industry, and damage to the micro-ecological balance of water bodies
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Embodiment 1
[0037] The screening and identification method of described Pseudomonas (Pseudomonas protegens) IAS03, the steps are as follows:
[0038](1) Weigh 10g of sediment from a pond in Guangzhou into a Erlenmeyer flask, add 90mL of sterilized water, 28°C, 160r / min, oscillate on a constant temperature shaker for 30min, let it stand for 5min, take the supernatant, and use the gradient dilution method Dilute the supernatant by 10 3 times, take 0.15mL of the diluted solution and spread it evenly on the beef extract peptone solid medium plate, and incubate at a constant temperature of 28°C for 16h. Beef extract peptone solid medium: 10g peptone, 3g beef extract, 5g sodium chloride, 15-20g agar, add water to 1000mL, pH7.0, sterilize at 121°C for 20min. The liquid medium of the same formula was without agar.
[0039] (2) Pick a single colony on the plate in step (1) and put it in beef extract peptone liquid medium, shake at 28°C at 160r / min for 8 hours on a constant temperature shaker, an...
Embodiment 2
[0056] Toxicity test of Pseudomonas protegens IAS03 strain to fish
[0057] (1) Experimental materials: 270 grass carp fry (about 20g in weight)
[0058] (2) Preparation of culture medium:
[0059] Beef extract-peptone liquid medium: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, pH 7.0-7.2, sterilized at 121°C for 20min;
[0060] Beef extract peptone solid medium: add 15-20g / L agar to the beef extract peptone liquid medium, and sterilize at 121°C for 20min.
[0061] (3) Preparation of injection bacterial solution:
[0062] Use an inoculation loop to take the preserved Pseudomonas IAS03 strain and streak it on the beef extract peptone solid medium plate. After culturing at 28°C for 16 hours, use an inoculation loop to pick a single colony and inoculate it in the beef extract peptone liquid medium. Shake culture at 160r / min for 12h at ℃, then centrifuge (7500rpm, 5 minutes), and wash with physiological saline repeatedly (7500rpm, 5 minutes) for three times, and then...
Embodiment 3
[0067] Inhibition test of 10ppm Pseudomonas IAS03 strain fermentation liquid on Aeromonas hydrophila and Aeromonas sobria
[0068] (1) Preparation of medium:
[0069] Beef extract-peptone liquid medium: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, pH 7.0, sterilized at 121°C for 20min;
[0070] Beef extract peptone solid medium: Add 15g / L agar to the beef extract peptone liquid medium, sterilize at 121°C for 20min, pour 50mL culture base into a Petri dish to make a solid medium plate.
[0071] LB liquid medium: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, pH value 7.2, sterilized at 121°C for 20min.
[0072] Use an inoculation loop to take Aeromonas hydrophila or Aeromonas temperatus preserved at -80°C, streak on the solid medium plate of beef extract peptone, cultivate at 28°C for 10 hours, and then use an inoculation loop to pick a single colony for inoculation In the beef extract peptone liquid medium, shake culture at 160r / min at 28°C for 1...
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