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A Virulent Vaccine Strain of Edwardsiella tarda and Its Application

A technology of slow Edwards strains, applied in the field of microbiology and immunology, can solve the problem of low protection of vaccines, and achieve the effect of improving safety, reducing losses and increasing biosafety risks

Active Publication Date: 2015-08-05
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

[0007] Generally speaking, compared with some other aquatic animal pathogenic bacteria, the difficulties encountered by researchers in the study of Edwardsiella tarda vaccines seem to be much greater. etc.), protein expression and purification technology (subunit vaccine), or gene knockout attenuation technology, in the vaccine research of Edwardsiella tarda, the problem of low protection of large and small vaccines has been encountered

Method used

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  • A Virulent Vaccine Strain of Edwardsiella tarda and Its Application
  • A Virulent Vaccine Strain of Edwardsiella tarda and Its Application
  • A Virulent Vaccine Strain of Edwardsiella tarda and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Isolation of Edwardsiella tarda strain 1101 from turbot

[0017] Pick out the dying turbot with typical symptoms of hemorrhagic disease, 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 (approx. 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, pH7.6-7.8), culture at 28°C for 24-48h, pick a single colony among the dominant colonies from the plate, streak the plate again, and carry out purification culture. The purified colony can be used for bacterial morphological characteristics, physiological and biochemical characteris...

Embodiment 2

[0018] Example 2: Cultivation of Edwardsiella tarda strain 1101

[0019] Edwardsiella tarda strain 1101 can be cultured in LB medium (tryptone 10g, yeast extract 5g, NaCl 10g, deionized water to 1L, pH7.0), TSB medium (Beijing Land Bridge Technology Co., Ltd.) or 2216E medium (according to the presence or absence of seawater in the medium components, the medium is divided into two types, one: same as Example 1; two: yeast extract 1g, tryptone 5g, FePO 4 0.1g, NaCl 34g, deionized water to 1L, pH7.6-7.8), and the preparation of the solid plate of the above medium needs to add 2% agar. 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 28-37°C for 24-48 hours to obtain monoclonal colonies on the plate. Take a single The colony was inoculated in the culture medium of the above medium, 28-37°C, 180r / min shaking flask culture to OD 600 If it is 0.5-0.6, the fresh...

Embodiment 3

[0020] Example 3: Physiological and biochemical characteristics of Edwardsiella tarda strain 1101

[0021] The physiological and biochemical tests of Edwardsiella tarda strain 1101 were identified according to the instructions of API20E (Mérieux, France) automatic identification card, and the results were analyzed with Apilab plus3.3.3 software. An additional oxidase test was added, and supplementary identification was carried out by determining the physiological and biochemical methods described in Dong Xiuzhu et al. (2001).

[0022] The test results showed that the oxidase was negative, and the identification results of the API20E automatic identification card are shown in Table 1, which showed that the strain 1101 was Edwardsiella tarda with a reliability of 99%.

[0023] Table 1 Biochemical identification results of Edwardsiella tarda 1101 strain API20E

[0024]

[0025] Note: "+" is positive, "-" is negative

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Abstract

The invention relates to an Edwardsiella tarda strain and an application method thereof. The Edwardsiella tarda strain is separated from a turbot adult fish body and is a wild strain with strong virulence, and the preservation number of the Edwardsiella tarda strain is CGMCC No.7197. Preparation modes of an antigen of the Edwardsiella tarda strain comprise any one or more than one of an inactivated thallus, a bacteruak ghost ingredient, an attenuated strain, a protective antigen, an antigen subunit and an expression product of an antigen determinant or an antigen gene expression carrier; the produced vaccine can be a single ingredient of the antigen prepared by utilizing the Edwardsiella tarda strain and can also be a combined vaccine produced by mixing the antigen prepared by utilizing the Edwardsiella tarda strain with antigens of other bacteria, and the prepared single or combined vaccine antigen is added with an adjuvant to produce the vaccine; and an inoculation mode of the vaccine in immunization application can adopt injection immunization, wound immunization, immersion bath immunization or oral administration immunization.

Description

technical field [0001] The invention relates to the field of microbiology and immunology of aquaculture animals, and in particular relates to an Edwardsiella tarda strain 1101 and its application. Background technique [0002] Edwardsiella tarda was first isolated from Anguilla japanica by Hoshina (1962). It is a Gram-negative short bacillus with a cell size of about 1 μm (diameter) × 2-3 μm (length). Oxygen belongs to the genus Edwardsiella of the Enterobacteriaceae, together with E.ictaluri and E.hoshinae. Edwardsiella tarda is pathogenic to a variety of fish, and it has been reported that it can infect fish such as: channel catfish (Ictalurus punctatus), largemouth bass (Micopterus salmoides), eel (A.japonica), mullet (Mugil cephalus ), bream (Evynnis japonica), tilapia (Tilapia nilotica), chinook salmon (Oncorhynchus tshawytscha), red snapper (Chrysophrys major), five bream (Seriola quinqueradiata), flounder (Paralichthys olivaceus), common carp ( Cyprinus carpio), sea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
Inventor 黄倢谢国驷王秀华隋虎辰杨春志边慧慧史成银张庆利许华刘莉
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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