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Triple inactivated vaccine, and preparation and application thereof

An inactivated vaccine and inactivated technology, applied in the field of aquatic vaccinology, can solve the problems of limited protective effect and achieve high immune protection, significant immune protective effect, and low cost

Inactive Publication Date: 2021-06-08
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 there have been reports in my country that vaccines for aquatic fish have obtained new veterinary drug certificates in recent years, these vaccines are mainly monovalent vaccines against one pathogen, and the protective effect is limited.

Method used

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  • Triple inactivated vaccine, and preparation and application thereof
  • Triple inactivated vaccine, and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Preparation of triple inactivated vaccine against Vibrio harveyi, Vibrio anguillarum, and Edwardsiella tarda.

[0022] step 1)

[0023] The Vibrio harveyi T4, Vibrio anguillarum C312, and Edwardsiella tarda TX1 are preserved in the General Microbiology Center CGMCC of the China Microbiological Culture Collection Management Committee. The address of the preservation unit is No. 13, North Zhongguancun, Haidian District, Beijing. The preservation numbers are : Vibrio harveyi T4 (CGMCC No.1985), Vibrio harveyi C312 (CGMCC No.6250), Edwardsiella tarda (CGMCC No.2330), named respectively as Vibrio harveyi (Vibrio harveyi), Vibrio harveyi Vibrio anguillarum Edwardsiella tarda.

[0024] Step 1) Proliferation and cultivation of bacteria. Vibrio harveyi, Vibrio anguillarum, and Edwardsiella tarda were resuscitated and inoculated on LB plates respectively, and a single colony was picked to continue passage twice, and a single colony was picked for Gram staining to observe its mo...

Embodiment 2

[0030] Immunization application of triple inactivated vaccine.

[0031] Step 1) Vaccine injection immunization. 1440 turbot (each weighing about 15g) were randomly divided into 6 groups, 240 in each group. These 6 groups were named as Groups A, B, C, D, E and F, respectively. Each fish in groups A-C was intraperitoneally injected with 100 μl of the triple vaccine mixture in step 3) of Example 1 above, and each fish in groups D-F was injected intraperitoneally with 100 ul of the control solution in step 3) of Example 1 above.

[0032] Step 2) Preparation of bacterial suspension for challenge. Single colonies of Vibrio harveyi, Vibrio anguillarum and Edwardsiella tarda were picked and inoculated in LB liquid medium respectively, and cultivated to OD 600 0.8-1.0, and then centrifuged (5000g, 4°C) for 5min. Bacteria were collected and suspended in PBS to a final concentration of Vibrio harveyi (5×10 7 cfu / ml), Vibrio anguillarum (5x10 7 cfu / ml), Edwardsiella tarda (1x10 4 ...

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Abstract

The invention relates to the field of aquatic product vaccinology, in particular to a vibrio harveyi, vibrio anguillarum and Edwardsiella tarda triple inactivated vaccine, and preparation and application thereof. The triple inactivated vaccine contains inactivated vibrio harveyi T4, vibrio anguillarum C312 and Edwardsiella tarda TX1. The highest relative percent survival (RPS) of the obtained inactivated vaccine for the vibrio harveyi, the vibrio anguillarum and the Edwardsiella tarda can be respectively 89.29%, 85.71% and 76.67%, and the RPS still can be about 50% in six months after immunization is carried out. The obtained vaccine has a simple preparation process and a low construction cost, does not have potential safety hazards, has a good immunization effect and can prevent and control three kinds of different-species pathogen infection through one-time immune inoculation.

Description

technical field [0001] The invention relates to the field of aquatic vaccinology, in particular to a triple inactivated vaccine of Vibrio harveyi, Vibrio anguillarum and Edwardsiella tarda and its preparation and application. Background technique [0002] my country is the world's largest aquaculture country, but with the expansion of industrial farming scale, aquatic diseases occur frequently, seriously restricting the development of aquaculture industry. At present, the control methods of marine aquaculture diseases in my country are still mainly based on the application of antibiotics and disinfectants, and the drug residues caused by them lead to food safety hazards, the increase of drug-resistant microorganisms, and ecological environment pollution. As a result, the aquatic application of antibiotics and disinfectants has been restricted in countries around the world. Vaccines, especially inactivated vaccines, are an internationally recognized green and pollution-free ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61P31/04
CPCA61K39/107A61K39/025A61P31/04A61K2039/521A61K2039/70A61K2039/552
Inventor 张健孙黎胡永华孙庆磊
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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