Novel edwardsiella attenuated vaccine strain as well as preparation method and application thereof

A technology of attenuated strains and uses, applied in the field of immunology, can solve the problems of endangering human health, declining disease resistance of the population, poor feeding of sick fish, etc.

Active Publication Date: 2021-09-17
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, chemically synthesized drugs or antibiotics are generally used in marine aquaculture production to control the large-scale outbreak of the disease, but there are many disadvantages in long-term drug use: 1) Sometimes the cause of fish diseases is complex, and there is blindness in a large number of drugs. At present, there is no targeted drug to treat; 2) The preventive effect of chemical drugs is very weak, and the sick fish eat poorly, and cannot ingest effective drug doses; 3) The drug...

Method used

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  • Novel edwardsiella attenuated vaccine strain as well as preparation method and application thereof
  • Novel edwardsiella attenuated vaccine strain as well as preparation method and application thereof
  • Novel edwardsiella attenuated vaccine strain as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0133]Embodiment 1, taking turbot as the toxicity test experiment of test animal

[0134] 1×10 4 CFU / tail and 1×10 7 Immune health turbot was intraperitoneally injected with CFU / tail dose, and the turbot used in the experiment was randomly divided into 4 groups, each group had 3 parallel tanks, 15 fish / tank. The prepared attenuated live vaccine and the like were immunized by intraperitoneal injection. The experimental group was injected with Edwardsiella EIB202 wild strain (WT), Edwardsiella 10-gene deletion strain (10Δ), and Edwardsiella 10-deficient overexpression esrB strain (10ΔesrB OE ), the control group was injected with an equal volume of sterile PBS buffer. The immunization period was 20 days, and the health status of the experimental animals was observed every day and the survival and death conditions were recorded.

[0135] Such as figure 1 As indicated, at 14 days of injection, 1×10 4 Nearly 70% of the CFU / tail WT group died, 10Δ and 10ΔesrB OE No fish died ...

Embodiment 2

[0137] Embodiment 2, the validity verification of animal level

[0138] 1. In vivo competition analysis experiment (CI) of turbot

[0139] Take 1ml of the secondary species of the related Edwardsiella strain cultured overnight at 37°C by shaking, centrifuge at 5000×g for 2 minutes to collect the bacteria, wash the bacteria three times with sterile PBS, and then dilute to 5×10 5 CFU / ml, mix each strain of EIB202 at a ratio of 1:1 as required. Inject the mixed bacterial solution into each fish with a challenge volume of 100 μl, 15 fish in each group. On the 5th day after challenge (DPI=5), the livers of 5 fishes in each group were grinded, and the dilution gradient plates were applied (to the DHL plate containing screening resistance and the blank DHL plate respectively). Incubate overnight at 37°C and count the colonies.

[0140] The result is as figure 2 As shown, the in vivo competition experiment was carried out with the EIB202Δp strain as a control, and it was found th...

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Abstract

The invention relates to a novel edwardsiella attenuated strain, wherein the expression of a target gene or a protein coded by the target gene is regulated and controlled. The edwardsiella attenuated strain can be used as a preventive drug for edwardsiella diseases, and has extremely obvious effect on inhibiting edwardsiella infection.

Description

technical field [0001] The invention belongs to the field of immunology, and more specifically, the invention relates to a novel attenuated vaccine strain of Edwardsiella, its preparation method and application. Background technique [0002] Edwardsiella can cause Edwardsiella, which is an important fish pathogen and can cause disease to more than 20 kinds of freshwater and seawater fish such as catfish, turbot and eel, causing gastrointestinal inflammation and hemorrhagic sepsis, resulting in A large number of fish die-offs seriously threaten the aquaculture industry. [0003] With the continuous expansion of aquaculture farming scale, large-scale outbreaks of aquaculture diseases often occur. Fish infectious diseases account for more than 20% of production losses every year, and large-scale disease outbreaks have become a major obstacle for the aquaculture industry. The main causes of fish infectious diseases include bacteria, parasites and viruses. [0004] Edwardsiell...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74A61K39/02A61P31/04C12R1/01
CPCC07K14/195C12N15/74A61K39/0208A61P31/04A61K2039/552A61K2039/522
Inventor 王启要殷开宇马佳宝马悦刘晓红刘琴张元兴徐荣静曲江波
Owner EAST CHINA UNIV OF SCI & TECH
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