Fish soaking vaccine multiple-combination adjuvant and application and using method thereof
A technology of soaking vaccines and adjuvants, which is applied in the field of multi-combination adjuvants for fish soaking vaccine immunization, which can solve the problems of low work efficiency, fish body damage, and few developed products, and achieves simple and convenient use, improved immune effect, Safe to use
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Embodiment 1
[0020] Embodiment 1: The application concentration of the present invention is respectively 0.1mg / L, 0.1mg / L, 50mg / L, 1mg / L NAC, TRITON X-100, saponin, 654-2 multi-combination adjuvant and inactivation Vibrio.Anguillarum Vaccine Immune to Soaked Flounder
[0021] 1 Material:
[0022] (1) Test fish and breeding management: choose healthy flounder with a weight of 5g, and temporarily raise them for 1 week before the test. The test seawater is stable at 20-23°C, PH8.1-8.2, salinity 30, and the compound bait is fed during the test. , fed twice a day, continuously inflated for 24 hours, running water culture.
[0023] (2) Vaccine preparation: inoculate V. anguillarum into 2216E liquid medium, incubate at 28°C for 24h; centrifuge at 4000rpm at 4°C for 20min to collect the bacteria; place the collected bacteria in sterilized PBS, pH 7.2, to prepare Vibrio suspension of 1.0×108cells / mL, inactivated by 0.5% formaldehyde at 28°C for 48 hours, then stored at 4°C; at the same time, the ...
Embodiment 2
[0034] Embodiment 2: The application concentration of the present invention is respectively 5mg / L, 5mg / L, 1mg / L, 30mg / L of NAC, TRITON X-100, saponin, 654-2 multi-combination adjuvant and inactivated point Aeromonas Vaccine Immune to Soaking Tilapia
[0035] 1 Material:
[0036] (1) Test fish and breeding management: select healthy tilapia with a weight of 3g, and temporarily raise them for 10 days before the test. The temperature of fresh water used in the test is 27-28°C, pH7.1-7.2. 2 times, 30% daily water change, 24 hours continuous inflation.
[0037] (2) Vaccine preparation: Inoculate Aeromonas punctatus into LB liquid medium, culture at 28°C for 24h; centrifuge at 4000rpm for 20min at 4°C, collect the bacteria; place the collected bacteria in sterilized normal saline, In pH 7.2, make a bacterial suspension of 1.0×109cells / mL, inactivate 0.5% formaldehyde at 28°C for 48 hours, then store it at 4°C; Cultivate at a constant temperature of 28°C for 24h and 48h, and confirm...
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