Fish soaking vaccine combination adjuvant and application and using method thereof
A technology for soaking vaccines and adjuvants, which is applied in the field of combined adjuvants for soaking vaccines for fish, can solve problems such as low work efficiency, fish body damage, and few products, and achieves the effects of simple and convenient use, improved immune effect, and safe use.
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Embodiment 1
[0016] Embodiment 1: application concentration of the present invention is respectively 1mg / L, 0.1mg / L, 60mg / L DTT, VD3, 654-2 combined adjuvant and inactivated Vibrio anguillarum (Vibrio.Anguillarum) vaccine to flounder immersion immunity
[0017] 1 Material:
[0018] (1) Test fish and breeding management: select healthy flounder with a weight of 5g, and raise them temporarily for 1 week before the test. The test seawater is stable at 20-23°C, pH8.1-8.2, salinity 30, and fed with compound bait during the test. , fed twice a day, continuously inflated for 24 hours, running water culture.
[0019] (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 inactivated bacte...
Embodiment 2
[0030] Embodiment 2: Application concentration of the present invention is respectively 50mg / L, 20mg / L, 1mg / L combined adjuvant of DTT, VD3, 654-2 and inactivated Aeromonas dot-shaped vaccine to tilapia immersion immunization
[0031] 1 Material:
[0032] (1) Test fish and breeding management: choose healthy tilapia with a weight of 3g, and temporarily raise them for 10 days before the test. Feed 2 times, change 30% of the water daily, and inflate continuously for 24 hours.
[0033] (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 that no colony grows.
[0034] (3) Immune adjuvants soaked: DTT, VD3, 65...
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