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Edwardsiella tarda attenuated live vaccine

A technology of slow Edwards and attenuated vaccines, applied in the field of attenuated vaccines, to achieve the effect of eliminating the danger of pathogens, clear attenuation mechanism and wide application range

Inactive Publication Date: 2009-01-14
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report on the development of attenuated live vaccine against Edwardsiella tarda using marker-free gene mutation technology at home and abroad

Method used

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  • Edwardsiella tarda attenuated live vaccine
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  • Edwardsiella tarda attenuated live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1 attenuated live vaccine preparation:

[0015] 1) TSB medium: tryptone 17.0g / L, soybean protein 3.0g / L, NaCl 5.0g / L, K 2 HPO42.5g / L, glucose 2.5g / L, pH 7.3.

[0016] Dissolve the above ingredients in 1000ml of distilled water, adjust the pH with 10% NaOH, sterilize at 121 lbs, 125°C for 15 minutes, cool down at room temperature for later use.

[0017] 2) TSA medium: tryptone 17.0g / L, soybean protein 3.0g / L, NaCl 5.0g / L, K 2 HPO42.5g / L, glucose 2.5g / L, agar powder 1.5g / L, pH 7.3.

[0018] Dissolve the above components in 1000ml of distilled water, adjust the pH with 10% NaOH, sterilize at 121 pounds, 125°C for 15 minutes, cool to 60°C to make a plate for use.

[0019] 3) PBS buffer: Na 2 HPO 4 2H 2 O 2.74g / L, NaH 2 PO 4 ·H 2 O 0.63g / L, the components were dissolved in 1000ml of distilled water, sterilized at 121 pounds, 125°C for 15 minutes, cooled and set at room temperature for later use.

[0020] Vaccine preparation: The seeds of MZLSE40esrB atte...

Embodiment 2

[0021] Embodiment 2 takes turbot (Scophthalmus maximus) as the median lethal dose LD of experimental animals 50 Determination of:

[0022] Healthy test fish with a body length of 8-10cm are temporarily raised in a 1000L aquarium, equipped with a flowing circulating water treatment and purification system, and the breeding water temperature is maintained at 16-18°C. After 4 weeks of temporary breeding, they were randomly divided into 20L experimental aquariums, and each group was set up with two parallel experimental aquariums, with 10 fish in each aquarium, and continued to be temporarily raised for 2 weeks. In the test aquarium, the water was changed once a day in the morning and afternoon, the amount of water changed was 1 / 2, and the water temperature was maintained at 16-18°C.

[0023] In the artificial infection test, each group of experimental fish was injected with different dilution concentrations of 10 2 -10 9 The attenuated vaccine strain and the wild-type bacteria...

Embodiment 3

[0027]Embodiment 3 takes the flounder (Paralichthys olivaceus) as the median lethal dose LD of the experimental animal 50 Determination of:

[0028] Healthy test fish with a body length of 8-10cm are temporarily kept in a 1000L aquarium, equipped with a flowing circulating water treatment and purification system, and the temperature of the breeding water is maintained at 18-20°C. After 4 weeks of temporary breeding, they were randomly divided into 20L experimental aquariums, and each group was set up with two parallel experimental aquariums, with 10 fish in each aquarium, and continued to be temporarily raised for 2 weeks. In the test aquarium, the water was changed once a day in the morning and afternoon, the amount of water change was 1 / 2, and the water temperature was maintained at 18-20°C.

[0029] In the artificial infection test, each group of experimental fish was injected with different dilution concentrations of 10 2 -10 9 The attenuated vaccine strain and the wild...

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Abstract

The present invention discloses a technique for preventing and curing the diseases of aquacultural animals, which relates to an attenuated vaccine for aquacultural bacterial diseases, in particular to an attenuated live vaccine for Edwardsiella tarda. The esrB gene of Edwardsiella tarda strain LSE40 loses an attenuated mutant strain, the Edwardsiella tarda mutant strain is MZLSE40esrB, which is preserved in China General Microbiological Culture Collection Center (CGMCC) and the accession number of which is CGMCC No.2087, and the mutant strain MZLSE40esrB does not contain exogenous antibiotic selectable markers and exogenous gene fragments. In respect to the wide type Edwardsiella tarda, the attenuated live vaccine has obviously low toxicity and immune protective rate and does not contain any exogenous antibiotic resistance marker and exogenous gene fragment. Experiments show that the attenuated live vaccine can effectively protect susceptible fishes from being infected with the pathogenic Edwardsiella tarda.

Description

technical field [0001] The invention relates to the disease prevention and control technology of aquaculture animals, and relates to attenuated vaccines for aquaculture bacterial diseases, in particular to a live attenuated vaccine for Edwardsiella tarda. Background technique [0002] Edwardsiella tarda is a Gram-negative pathogen that infects a wide range of hosts and can infect a variety of lower and higher animals. In aquaculture animals, the bacterium is an important pathogenic bacterium of fish, amphibians and reptiles. The Edwardsiella disease caused by it is a common infectious disease in aquaculture fish, which is very harmful to the health of farmed animals and the aquaculture industry. In my country, pathogenic Edwardsiella has been isolated from various animals such as bullfrog, soft-shelled turtle, eel, flounder, turbot, and red sea bream. [0003] Due to the use of various antibiotics for disease prevention and control in the production process, more and more ...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61K47/04A61P31/04C12N1/20
Inventor 莫照兰茅云翔肖鹏李杰王波杨佳银
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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