Vibrio anguilarum oil oral-emulsified bivalent vaccine and preparation thereof
A technology for oral vaccines and vaccines, applied in antibacterial drugs, emulsion delivery, bacterial antigen components, etc., can solve the problems of narrow immune protection spectrum and high material cost, achieve low material cost, broaden vaccine protection spectrum, and increase immune synergy The effect of action
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Embodiment 1
[0040] Listonella MacDonellet Collwell's Listonella anguillarum Vibrioanguillarum 01 serotype subtype strain M3 is preserved in the General Microorganism Center No. 1870 of China Committee for the Collection of Microorganisms.
[0041] Listonella MacDonellet Collwell's Listonella anguillarum Vibrioanguillarum 01 serotype subtype strain SMP1 is preserved in the General Microbiology Center No. 1871 of China Committee for the Collection of Microorganisms.
[0042] Morphological characteristics of strain M3:
[0043] The colony of the strain in 2216E medium for 24 hours was round, translucent, with smooth surface and neat edges, with a diameter of about 1.5mm; the colony in TCBS medium was yellow, with a diameter of about 3mm; Gram staining was negative and short rod-shaped. Can move. There is a very long polar single flagella and periphyme, and the size of the bacteria is 0.8~1.0×1.5~1.7μm (see figure 1 ).
[0044] Strain M3 is positive, sensitive to Vibrio inhibitor 0 / 129 (2,...
Embodiment 2
[0053] The difference from Example 1 is:
[0054] 1) Prepare the water phase of the emulsified vaccine: the concentration of formaldehyde inactivation after the rejuvenation culture is 4×10 9 cells / ml of Listonia eel 01 serotype two subtype strains were added to Tween-80, stirred and mixed to obtain the aqueous phase of the emulsified vaccine; the bacterial suspension was 18.8ml according to the volume percentage of 94%. , Tween-80 takes 1.2ml by volume percentage 6%.
[0055] 2) Prepare the oil phase of the emulsified vaccine: prepare the oil phase of the emulsified vaccine: get 27.6ml No. 10 white oil by volume percentage 92%, get 2.4ml Span-80 by volume percentage 8%, get by the total mass volume ratio of the oil phase 1.5% 0.45g aluminum stearate is mixed, heated to make the aluminum stearate dissolve completely, and sterilized to prepare the oil phase of the emulsified vaccine;
[0056] 3) Preparation of emulsified vaccine: 20ml of water phase was slowly added to 40ml o...
Embodiment 3
[0058] The difference from Example 1 is:
[0059] 1) Prepare the water phase of the emulsified vaccine: the concentration of formaldehyde inactivation after the rejuvenation culture is 6×10 9 cells / ml of Listonia eel 01 serotype two subtype strains were added to Tween-80, stirred and mixed to obtain the aqueous phase of the emulsified vaccine; wherein the bacterial suspension was 19.6ml according to the volume percentage of 98%. , Tween-80 takes 0.4ml by volume percentage 2%.
[0060] 2) Prepare the oil phase of the emulsified vaccine: get 28.8ml No. 10 white oil by volume percentage 92%, get 1.2ml Span-80 by volume percentage 4%, get 0.75g aluminum stearate by volume ratio 2.5% of the total mass of the oil phase , mix them, heat to make the aluminum stearate dissolve completely, and sterilize to prepare the oil phase of the emulsified vaccine;
[0061] 3) Preparation of emulsified vaccine: 20ml of water phase was slowly added to 36ml of oil phase at a volume percentage of 1...
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