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Vibrio anguillarum exuviae vaccine and method of preparing the same

A technology of vibrio eel and empty shells, which is applied in the field of bacterial empty shell vaccines for aquaculture fish, which can solve the problems of unreported aquaculture fish and achieve significant protective effects, broad development prospects, and low production costs

Inactive Publication Date: 2008-01-23
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although bacterial empty-shell vaccines have been extensively studied and applied in human and veterinary medicine, only Edwardsiella empty-shell vaccines have been preliminarily studied in aquatic products, but no reports have been found in aquaculture fish

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1) Selection of recipient bacteria: Vibrio anguillaris strain M3 in flounder in which vibriosis broke out in seawater fish farms in the Yellow Sea of ​​my country was selected as the recipient bacteria.

[0019] 2) Construction of a plasmid vector system: subcloning the nucleotide fragments containing the bacteriophage PhiX174 cleavage gene E and its expression regulation system on the pGEMT vector;

[0020] 3) Transformation: The vector constructed in step 2) was transformed into a recipient strain of Vibrio anguillarum, and stored in glycerol at -80°C until use.

[0021] 4) Cultivate:

[0022] ① Pre-cultivation: Inoculate engineered Vibrio anguillarum preserved in glycerol into TSB medium containing ampicillin, the inoculum volume of each bottle is 10ul, in which: each bottle contains 10ml TSB medium and 500ug of ampicillin, each liter of TSB medium Contains: tryptone 17g, soybean peptone 5g, NaCl 5g, K 2 HPO 4 2.5g, glucose 2.5g, pH 7.6; placed in a shaker flask a...

Embodiment 2

[0027] 1) Selection of recipient bacteria: Vibrio anguillarum strain SMP1 in turbot that broke out in vibriosis outbreaks in seawater fish farms in the Yellow Sea of ​​my country was selected as the recipient bacteria.

[0028] 2) Construction of a plasmid vector system: subcloning the nucleotide fragments containing the bacteriophage PhiX174 cleavage gene E and its expression regulation system on the pGEMT vector;

[0029] 3) Transformation: The vector constructed in step 2) was transformed into a recipient strain of Vibrio anguillarum, and stored in glycerol at -80°C until use.

[0030] 4) Cultivate:

[0031] ①Pre-cultivation: inoculate the engineered Vibrio anguillarum preserved in glycerol into TSB medium containing ampicillin, with an inoculation volume of 30ul per bottle, of which: each bottle contains 15ml TSB medium and 500ug of ampicillin; placed at 200rpm, 28°C in shake flasks for 14 hours.

[0032] ②Empty shell preparation: first transfer the 14-hour-cultured stra...

Embodiment 3

[0036] 1) Selection of recipient bacteria: Vibrio anguillarum strain SMP in turbot that broke out of vibriosis in seawater fish farms in the Yellow Sea of ​​my country was selected as the recipient bacteria.

[0037] 2) Construction of a plasmid vector system: subcloning the nucleotide fragments containing the cleavage gene E of the phage PhiX174 and its expression regulation system on the pGEMT vector.

[0038] 3) Transformation: The vector constructed in step 2) was transformed into a recipient strain of Vibrio anguillarum, and stored in glycerol at -80°C until use.

[0039] 4) Cultivate:

[0040] ① Pre-cultivation: Inoculate engineered Vibrio anguillarum preserved in glycerol into TSB medium containing ampicillin, the inoculum volume of each bottle is 50ul, wherein: each bottle contains 20ml TSB medium and 1mg ampicillin; place at 200rpm, 30℃ in shake flasks for 16 hours.

[0041] ②Empty shell preparation: first transfer the 16-hour-cultured strain to the TSB medium conta...

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PUM

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Abstract

The invention relates to a bacterium coreless vaccine used for the fish aquaculture, in particular to a Vibiro anguillarum coreless vaccine and the preparation method thereof. The Vibiro anguillarum is an engineering bacteria construction carrier and then turn, foster, filter and purify the carrier to prepare the coreless vaccine. The adoption of the Vibiro anguillarum coreless vaccine retains the complete antigenic determinant of the Vibiro anguillarum cell membrane, overcomes the defect of antigenic damage caused by the traditional physical and chemical vaccine inactivation way. By immunizing the fish by oral administration and injection, the invention can better protect the fish compared with the traditional inactive cell vaccine. In addition, the invention has commercial development value due to low cost and broad prospect in developing the polyvalent vaccine and preventing various diseases as a vaccine carrier.

Description

technical field [0001] The invention relates to a bacterial empty shell vaccine for aquaculture fish, in particular to an empty shell vaccine of Vibiro anguillarum and a preparation method thereof. technical background [0002] Vibrio anguillarum is one of the important pathogens that cause sepsis in aquaculture animals, and the vibriosis caused by it is the main disease of industrially farmed fish in my country. The prevention and treatment of vibriosis mainly consists of drug therapy and vaccine immunization. Drug treatment involves the use of antibiotics and chemical drugs, which is likely to cause drug residues in aquatic products and bacterial resistance, causing harm to the environment and human health. Therefore, the use of antibiotics is greatly restricted worldwide. Vaccine immunization is superior to drug treatment such as antibiotics in terms of specificity, safety and protective effect, and is an effective means of disease prevention and control. There are dead...

Claims

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Application Information

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IPC IPC(8): A61K39/106A61P31/04
Inventor 莫照兰茅云翔邹玉霞肖鹏王波叶旭红李杰徐永立张培军
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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