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Pseudomonas strain and application thereof

A technology for Pseudomonas and Aeromonas, applied in the field of microorganisms, can solve the problems of polluted water environment, production of drug-resistant pathogens, hemorrhagic sepsis, etc.

Active Publication Date: 2016-02-24
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Aeromonas hydrophila (Aeromonas hydrophila) and temperate aeromonas (Aeromonassobria) exist in the aquaculture water environment, and often cause almost all freshwater cultured fish to suffer from hemorrhagic sepsis, with high morbidity and mortality. Huge economic losses caused by the farming industry
In the past, antibiotics were mainly used to prevent and treat the disease, and topical disinfectants were used to kill pathogenic bacteria in water bodies. In the long run, this would not only produce drug residues, induce drug-resistant pathogenic bacteria and pollute the water environment, but also disinfectants would not selectively kill beneficial bacteria in water. Destroying the micro-ecological balance of the water body, laying a hidden danger for the subsequent outbreak of bacterial sepsis

Method used

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  • Pseudomonas strain and application thereof
  • Pseudomonas strain and application thereof
  • Pseudomonas strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The screening and identification method of described pseudomonas (Pseudomonasprotegens) 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, shake on a constant temperature shaker at 28°C, 160r / min for 30min, let stand for 5min, take the supernatant, and use the gradient dilution method Dilute the supernatant by 10 3times, 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, and wait...

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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Abstract

The invention discloses a pseudomonas strain and application thereof. The strain is pseudomonas protegens IAS03, and is preserved in the China center for type culture collection on November 27th, 2015, and the preservation number is CCTCC NO:M 2015711. The strain is screened from pond bottom sludge and then a backwater body is used, so that ecological safety risks do not exist, and biological safety regulations are conformed. The strain has the specific antimicrobial function on aeromonas hydrophila and aeromonas sobria, has no inhibition effect on other bacteria and has no pathogenicity on fishes. Only one strain can be used for inhibiting aeromonas hydrophila and / or aeromonas sobria. Compared with traditional antibiotic and sterilizing agent prevention measures, the strain has the advantages of being safe, environmentally friendly and efficient. The strain can be developed into a microecological preparation to control aeromonas sobria and aeromonas hydrophila in the breeding environment and prevent hemorrhagic septicemia caused when the fishes are infected with aeromonas sobria and aeromonas hydrophila.

Description

technical field [0001] The invention belongs to the field of microorganisms, and relates to a pseudomonas strain, in particular to a pseudomonas strain and application thereof. Background technique [0002] Aeromonas hydrophila (Aeromonas hydrophila) and temperate aeromonas (Aeromonassobria) exist in the aquaculture water environment, and often cause almost all freshwater cultured fish to suffer from hemorrhagic sepsis, with high morbidity and mortality. The aquaculture industry has caused significant economic losses. In the past, antibiotics were mainly used to prevent and treat the disease, and topical disinfectants were used to kill pathogenic bacteria in water bodies. In the long run, this will not only produce drug residues, induce drug-resistant pathogens and pollute the water environment, but also disinfectants will not selectively kill beneficial bacteria in water. It destroyed the micro-ecological balance of the water body and laid a hidden danger for the subsequen...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/38
Inventor 张其中王飞飞
Owner JINAN UNIVERSITY
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