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Prepn and usage of outer membrane protein subunit vaccine of seawater fish morbid vibrio

A subunit vaccine and outer membrane protein technology, applied in the field of preparation and use of marine fish pathogenic Vibrio outer membrane protein subunit vaccine, can solve the problems of large dosage, easy rebound, poor immune effect, etc. Stable specificity and easy-to-use effects

Inactive Publication Date: 2007-08-22
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Vibrio anguillarum vaccine has poor immune effect due to the virulence that is easy to rebound, the dosage is large, and the body’s immune response level is low.
The minimal vibrio conjugated subunit vaccine has a relatively specific anti-vibrio specificity in aquaculture production, and has poor defense against diseases caused by other vibrio pathogenic bacteria, which has obvious limitations in production and application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Vibrio alginolyticus and Vibrio harveyi were first activated with trypticase soytone liquid medium (TSB), cultured at 28°C with shaking at 150r / min for 18h, and then 1mL of the bacterial solution was applied to the culture medium containing 2% NaCl and The tryptone soy agar plate medium (TSA) with an initial pH of 7.2 was cultured statically at 28° C. for 18 hours. Then, the outer membrane proteins of Vibrio alginolyticus and Vibrio harveyi were extracted respectively. The method is as follows: wash the colonies separately with 0.01mol / L PBS with a pH of 7.2, centrifuge at 7000r / min at 4°C for 10 minutes, repeat the washing twice, take the sludge and suspend it in PBS, and ultrasonically break it in an ice bath for 7 minutes, at 4°C, Centrifuge at 7000r / min for 15min, remove the large bacterial fragments, and centrifuge at 35000r / min at 4°C for 1h. Collect the precipitate with the above PBS, which is the outer membrane protein extract, and store at -20°C. Next, the ou...

Embodiment 2

[0034] Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus and Vibrio anguillarum were first activated with trypticase soytone liquid medium (TSB), cultured at 28°C with shaking at 150r / min for 18h, and then 1 mL of the bacterial liquid was taken separately Spread on tryptone soy agar plate medium (TSA) containing 2% NaCl and the initial pH of 7.2, and culture at 28° C. for 18 hours. Then, the outer membrane proteins of Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus and Vibrio anguillarum were extracted respectively. The method is as follows: wash the colonies separately with 0.01mol / L PBS with a pH of 7.2, centrifuge at 7000r / min at 4°C for 10 minutes, repeat the washing twice, take the sludge and suspend it in PBS, and ultrasonically break it in an ice bath for 7 minutes, at 4°C, Centrifuge at 7000r / min for 15min, remove the large bacterial fragments, and centrifuge at 35000r / min at 4°C for 1h. Collect the precipitate with the above PBS, which is the outer memb...

Embodiment 3

[0036]Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus, Vibrio anguillarum, Vibrio parahaemolyticus and Vibrio mimicus were respectively activated by trypticase soy peptone liquid medium (TSB), at 28°C, shaking at 150r / min After culturing for 18 hours, 1 mL of the bacterial solution was spread on tryptone soy agar plate medium (TSA) containing 2% NaCl and an initial pH of 7.2, and cultured statically at 28° C. for 18 hours. Then, the outer membrane proteins of Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus, Vibrio anguillarum, Vibrio parahaemolyticus and Vibrio mimicus were extracted respectively. The method is as follows: wash the colonies separately with 0.01mol / L PBS with a pH of 7.2, centrifuge at 7000r / min at 4°C for 10 minutes, repeat the washing twice, take the sludge and suspend it in PBS, and ultrasonically break it in an ice bath for 7 minutes, at 4°C, Centrifuge at 7000r / min for 15min, remove the large bacterial fragments, and centrifuge at 35000r / mi...

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Abstract

The present invention is preparation and usage of outer membrane protein subunit morbid vibrio vaccine for seawater fish. The vaccine is prepared through extracting two or more of Vibrio alginolyticus, Vibrio Hrveyi, Vibrio vulnificus, Vibrio parahaemolyticus, etc, and adding immunostimulating complex. The vaccine preparing process includes culturing vibrio, extracting outer membrane protein and preparing vaccine. The vaccine containing partial pathogen and no infecting component has stable immunological specificity and no hidden danger of restoring toxici. The vaccine is used in preventing fishes' skin ulcer, eyeball turbidity and other vibrio caused diseases and may be used widely for immunizing seawater fish.

Description

Technical field: [0001] The invention belongs to a method for preparing and using an outer membrane protein subunit vaccine of vibriosis in seawater fish. Background technique: [0002] In recent years, with the rapid development of the mariculture industry, the scale of marine fish cage culture has also continued to expand. Due to the increase in the number of cages in the culture sea area and the increase in the density of culture, the environment of the culture waters has gradually deteriorated, and the occurrence of diseases has become more and more frequent. Bacteria Diseases have become one of the most common and most harmful diseases in aquaculture, often leading to a large number of deaths of farmed fish, among which Vibrosis (Vibrosis) is the most common and most harmful disease in marine fish farming. The aquaculture industry causes huge economic losses. [0003] For a long time, traditional control methods have generally used chemical drugs and antibiotics. Exten...

Claims

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Application Information

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IPC IPC(8): A61K39/106A61K47/24A61P31/04
Inventor 吴灶和庞欢瑛姚绍云简纪常
Owner GUANGDONG OCEAN UNIVERSITY
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