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Pseudomonas anguilliseptica vaccine and applications thereof
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A technology for pseudomonas and vaccines, applied in the fields of microbiology and immunology, can solve problems such as poor stability and poor vaccine adaptability, and achieve the effects of reducing losses, improving safety, and good immune protection.
Inactive Publication Date: 2018-02-16
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology
[0004] In order to solve the problem of poor adaptability and poor stability of existing vaccines, we propose a Pseudomonas eeliseptic vaccine and its application
Method used
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Embodiment 1
[0024] Isolation of Pseudomonas anguillarum septicum strain PLSH160511 from diseased star flounder
[0025] Dying flounder with symptoms typical of haemorrhagic diseases were selected (see figure 1 ), disinfect the body surface of the diseased fish with 70% alcoholcotton balls, cut open the abdominal cavity with sterile scissors, and take out small pieces of liver, kidney, spleen and other tissues (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 40.1g, dilute seawater to 1L, pH 7.6—7.8), culture at 20°C for 2 weeks, 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, physiolog...
Embodiment 2
[0027] Cultivation of Pseudomonas septicum strain PLSH160511
[0028] Pseudomonas anguli septica strain PLSH160511 can be cultured in TSB+MgCl 2 Medium (tryptone soybean broth 30g, NaCl 10g, MgCl 2 ·6H 2 O 0.2g, deionized water to 1L, pH 7.0), or 2216E medium (yeast extract 1g, tryptone 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 in addition. The culture method of the strain 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 16-28°C for 2 weeks to obtain monoclonal colonies on the plate. Take a single colony Inoculate in the culture medium of the above-mentioned culture medium, culture at 16-28°C, shake the flask at 180r / min until OD 600 If it is 0.5-0.6, the fresh bacterial liquid of the strain can be obtained.
Embodiment 3
[0030] Physiological and biochemical characteristics of Pseudomonas anguli septica strain PLSH160511
[0031] Physiological and biochemical tests of Pseudomonas septicum strain PLSH160511 were identified with reference to the API32E (Mérieux, France) automatic identification card instructions, and the results were analyzed with Apilab plus 3.3.3 software, showing that the strain was Pseudomonas, Uncommon identification results.
[0032] Table 1 Biochemical identification results of API32E of Pseudomonas eel septica PLSH160511 strain
[0033]
[0034]
[0035] Note: "+" means positive; "-" means negative
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Abstract
The invention discloses a strain PLSH160511 of pseudomonas anguilliseptica. The strain is separated from verasper variegates cultured in China. The strain is strongly pathogenic for verasper variegates and turbot. The preservation number of the strain is CCTCC M 2017269. In an experiment, an immune protective rate (80% or more) can be obtained by using an inactivated emulsified vaccine prepared from the strain. The immune protection duration is 6 months or more. Compared with a hydration vaccine, the immune protective rate is higher, the immune protective rate of the hydration vaccine is 60%;the immune protection duration is longer, and the immune protection duration of the hydration vaccine is 3 months. The vaccine prepared from the strain PLSH160511 of pseudomonas anguilliseptica has immune protective property of pseudomonas anguilliseptica. The strain PLSH160511 can be used to produce vaccines and applied to vaccine development.
Description
technical field [0001] The invention relates to the field of microbiology and immunology of aquaculture animals, in particular to a vaccine for Pseudomonas eel septica and its application. Background technique [0002] Pseudomonas anguilliseptica is an important pathogenic bacterium of marine and brackish water cultured fish. It was originally isolated from the Japanese eel (Anguilla japonica) where the "red spot disease" broke out. It is a Gram-negative short bacillus with a cell size of about 1 μm (diameter) × 2-3 μm (length). Pseudomonas eel septica is pathogenic to a variety of freshwater and brackish water fish, and its optimum infection temperature is 17-21°C, and it frequently occurs in winter when the water temperature is low. Among the pathogenic Pseudomonas, Pseudomonas anguillae is the greatest threat to marine fish, and cases of Pseudomonas anguillaris infection have been found in a variety of cultured fish, including European eel (Anguilla anguilla), black sea ...
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