High-efficiency non-injection type transmission method for live vaccine
An injection-type vaccine technology, applied in the field of vaccinology, can solve the problem of low immune effect and achieve the effect of simple application, low price and high protection rate
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Embodiment 1
[0018] Routine single injection delivery of vaccines
[0019] Step 1) Immunization injection of vaccine. Cultivate Edwardsiella tarda vaccine strain 15947 (purchased from ATCC, USA) at 28°C in LB medium to OD 600 0.8-1, and then centrifuged (5000g, 4°C) for 10min. Collect the cells and suspend them in PBS to a final concentration of 1x10 8 cfu / ml, that is, 15947 suspension. 50 flounder were randomly divided into 2 groups (25 in each group), named A and B respectively. The fish in groups A and B were injected intraperitoneally with 15947 suspension or PBS (control group), and the injection volume of each fish was 100ul.
[0020] The LB composition is by weight percentage: 1.0% peptone, 0.5% yeast powder, 1.0% sodium chloride, 97.5% distilled water; the PBS composition is by weight percentage: 0.8% NaCl, 0.02% KCl, 0.358% Na 2 HPO 4 .12H 2 O, 0.024% NaH 2 PO 4 , 98.798% distilled water.
[0021] Step 2) Detection of the immune protection effect of the vaccine. Cultiva...
Embodiment 2
[0024] oral delivery of vaccines
[0025] Step 1) Preparation of vaccine sodium alginate microsphere feed and control sodium alginate microsphere feed. Cultivate Edwardsiella tarda vaccine strain 15947 to OD in LB medium at 28 °C 600 0.8-1, and then centrifuged (5000g, 4°C) for 10min. Collect the cells and suspend them in normal seawater to a final concentration of 1x10 9 cfu / ml, that is, 15947 suspension. Mix 50ml of sodium alginate (3%) with 30ml of 15947 suspension or PBS (control), then add 2000ml of paraffin, 10ml of S-80 emulsifier (all purchased from Sigma, USA) and 50ml of calcium chloride (0.15M) ( purchased from Shanghai Sangon Company); the mixed solution containing 15947 and PBS contrast was centrifuged (1000g, 10min) respectively, and the precipitate was collected; ) and mixed with F(II)-26. Twin-screw extruder granulator (South China University of Technology, Guangzhou). Processed into pellet feed (1.5mm×2.0mm), namely vaccine sodium alginate microsphere feed...
Embodiment 3
[0029] Immersion transfer of vaccines
[0030] Step 1) Preparation of vaccine soaking solution. Grow vaccine strain 15947 to OD in LB medium at 28 °C 600 0.8-1, and then centrifuged (5000g, 4°C) for 10min. Collect the cells and suspend them in normal seawater to a final concentration of 1x10 8 cfu / ml is the vaccine soaking solution.
[0031] Step 2) Immunization delivery of the vaccine. Fifty flounder were randomly divided into 2 groups (25 in each group), named E (sample group) and F (control group). Soak the fish of group E in the vaccine soaking solution of the above step 1) for 1 hour, and soak the fish of group F in ordinary sea water for 1 hour.
[0032] Step 3) Detection of the immune protection effect of the vaccine. Cultivation of Edwardsiella tarda TX1 to OD in LB medium 600 0.7, and then centrifuged (5000g, 4°C, 10min). Collect the cells and suspend them in PBS to a final concentration of 5x10 8 cfu / ml is the suspension of Edwardsiella tarda. On the 30th d...
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