Edwardsiella tarda attenuated live vaccine
A technology for delayed Edwards, attenuated vaccines, applied in the field of attenuated vaccines
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Embodiment 1
[0015] Embodiment 1 attenuated live vaccine preparation:
[0016] 1) TSB medium: tryptone 17.0g / L, soybean protein 3.0g / L, NaCl 5.0g / L, K 2 HP42.5g / L, glucose 2.5g / L, pH7.3.
[0017] 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.
[0018] 2) TSA medium: tryptone 17.0g / L, soybean protein 3.0g / L, NaCl 5.0g / L, K 2 HP42.5g / L, glucose 2.5g / L, agar powder 1.5g / L, pH7.3.
[0019] 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.
[0020] 3) PBS buffer: Na 2 HPO 4 2H 2 O2.74g / L, NaH 2 PO 4 ·H 2 0.63g / L, said composition is dissolved in 1000ml distilled water, in 121 pounds, 125 ℃ of sterilization 15 minutes, cooling puts room temperature for subsequent use.
[0021] Vaccine preparation: The seeds of MZLSE40esrB attenuated v...
Embodiment 2
[0022] Embodiment 2 takes turbot (Scophthamus maximus) as the median lethal dose LD of experimental animals 50 Determination of:
[0023] 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 change was 1 / 2, and the water temperature was maintained at 16-18°C.
[0024] 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 bacterial ...
Embodiment 3
[0028] Embodiment 3 takes the flounder (Paralichthys olivaceus) as the median lethal dose LD of the experimental animal 50 Determination of:
[0029] 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.
[0030] 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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