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

Inactive Publication Date: 2018-12-18
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] To address the lack of existing flounder vaccines, we propose a Heterovibrio salmonicida vaccine and its application

Method used

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  • Alivibrio salmonicida vaccine and application thereof
  • Alivibrio salmonicida vaccine and application thereof
  • Alivibrio salmonicida vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

According to the invention, an Alivibrio salmonicida strain is separated from Paralichthys olivaceus cultured in China. The strain has strong pathogenicity to Paralichthys olivaceus and Atlantic salmon and is preserved under the accession number of CCTCC M 2018086. In experiments, an inactivated vaccine prepared from the strain can realize an immune protective rate of 80% or more, and the durationof the immunoprotection lasts for 6 months or more; so it is indicated that the vaccine prepared from Alivibrio salmonicida PO-1 has the immunoprotective effect on Alivibrio salmonicida. The Alivibrio salmonicida strain is applicable to preparation of the vaccine and has the application prospects of vaccine development.

Description

technical field [0001] The invention relates to the field of microbiology and immunology of aquaculture animals, in particular to an Alivibriosalmonicida vaccine and application thereof. Background technique [0002] Alivibriosalmonicida is an important pathogenic bacterium of marine fish, which can cause cold-water vibriosis in farmed Atlantic salmon (Salmo salar), Atlantic cod (Gadus morhua) and rainbow trout (Oncorhynchus mykiss). The resulting mortality of farmed fish reaches 50-90%. In the history of Norwegian aquaculture, more than 80% of the disease losses have been related to I. salmonicida. I. salmonicida is a Gram-negative short bacillus with a cell size of about 1 μm (diameter)×2-3 μm (length). At present, there is no case report of this bacteria infection in our country. I. salmonicida is widely present in seawater environment. It occurs frequently in winter when the water temperature is low. It occurs mostly in fish cultured in cages and coffered ponds. The i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61P31/04C12N1/20C12R1/63
CPCA61K39/107A61K2039/542A61P31/04C12N1/205C12R2001/63
Inventor 李杰莫照兰刘长琳李贵阳陈四清
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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