Alivibrio salmonicida vaccine and application thereof
A vaccine and vibrio technology, applied in the field of microbiology and immunology of aquaculture animals, can solve problems such as lack of vaccines, and achieve the effects of reducing losses, good immune protection, and improving safety
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Embodiment 1
[0023] Isolation of Heterovibrio salmonicida Strain PO-1 from Diseased Flounder
[0024] Pick out dying flounder with typical symptoms of hemorrhagic disease (see figure 1 ), disinfect the body surface of the diseased fish with 70% alcohol cotton balls, cut open the abdominal cavity with sterile scissors and take out small pieces of liver, kidney, spleen and other tissues of the diseased fish (about 0.2×0.2×0.2cm 3 ) in a 1mL sterile centrifuge tube, rinsed repeatedly with sterile PBS (pH7.2) on the ultra-clean workbench, dipped the above lesion tissue with an inoculation loop, and inoculated it in seawater 2216E medium supplemented with 2% agar ( Yeast extract 1g, tryptone 5g, FePO 4 0.1g, dilute seawater to 1L, pH 7.6-7.8), culture at 16°C for 5 days, pick a single colony among the dominant colonies from the plate, streak the plate again, and carry out purification culture. The purified bacterial colony can carry out bacterial morphological characteristics, physiological a...
Embodiment 2
[0026] Cultivation of I. salmonicida Strain PO-1
[0027] I. salmonicida strain PO-1 can be cultured in TSB medium (tryptone soybean broth 30g, NaCl 10g, deionized water to 1L, pH 7.0), or 2216E medium (yeast extract 1g, tryptone Peptone 5g, FePO 4 0.1g, seawater to 1L, pH 7.6-7.8), the preparation of the solid plate of the above medium needs to add 2% agar additionally. The strain cultivation method is as follows: pick a small amount of strains stored at -80°C and streak on the solid plate of the above-mentioned medium, and culture it at 10-25°C for 1 day to obtain monoclonal colonies on the plate. Take a single colony Inoculate in the culture solution of the above-mentioned medium, shake the flask at 10-25°C, 180r / min, and cultivate to OD 600 If it is 0.5-0.6, the fresh bacterial liquid of the strain can be obtained.
Embodiment 3
[0029] Physiological and biochemical characteristics of I. salmonicida strain PO-1
[0030] Physiological and biochemical tests of Alt. salmonicida strain PO-1 were identified by referring to the instructions of Biolog (BIOLOG, USA) automatic plate, and the results were consistent with Al. salmonicida.
[0031] Table 1 BIOLOG biochemical identification results of I. salmonicida PO-1 strain
[0032]
[0033]
[0034]Note: "+" means positive; "-" means negative; "w" means weak positive reaction.
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