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Preparation technique of Vibrio vulnificus whole-cell immunity stimulated compound vaccine and use thereof in aquatic animal

A technique for Vibrio vulnificus and immune stimulation, applied in the field of preparation of pathogenic bacteria inactivated vaccines

Inactive Publication Date: 2009-02-11
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The research on fermentation technology mainly focuses on industrial microorganisms such as Escherichia coli, and there has been no report on the fermentation technology of Vibrio; the research reports on ISCOMs of aquatic animal pathogens can only be found in the ISCOMs research of Aeromonas hydrophila in our laboratory, and Vibrio vulnificus The research on the preparation process of ISCOM vaccine has not been reported in China

Method used

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  • Preparation technique of Vibrio vulnificus whole-cell immunity stimulated compound vaccine and use thereof in aquatic animal
  • Preparation technique of Vibrio vulnificus whole-cell immunity stimulated compound vaccine and use thereof in aquatic animal

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Embodiment Construction

[0007] Concrete processing steps of the present invention are as follows:

[0008] (1) Pre-amplification and expansion culture of Vibrio vulnificus

[0009] Select recognized TSB medium and improved TSB medium; inoculate Vibrio vulnificus strains that have passed routine testing into a 250ml Erlenmeyer flask containing 50ml TSB of TSB bacterial culture, keep the temperature at 25-31°C, and rotate at 100-200 Under the condition of vibration per minute for 14-18 hours, the culture solution is used as the seed solution; 0.5-10% pre-amplified culture is inoculated into the fresh autoclaved modified TSB culture solution in the fermenter for large-scale cultivation 36-60 hours; the composition of the improved TSB culture fluid is: glucose 0.18%-0.22% (W / V is the volume weight percentage of water, the same below.), tryptone 1.2%, 1.8% (W / V ), soybean peptone 0.2%-0.6% (W / V), corn steep liquor 0.2%-0.6% (V / V), dipotassium hydrogen phosphate 0.15%-0.25% (W / V), sodium chloride 1.0%-2.0...

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Abstract

The invention relates to a preparation technology of vibrio vulnificus whole-bacteria immunity stimulated compound vaccine and the application thereof in aquatic animals. The technology comprises the following steps: (1) pre-amplification and expansion cultivation of the vibrio vulnificus is carried out, (2) blanching is imposed on the vibrio vulnificus in zymogen liquid cultivated, (3) non-viable bacteria inspection is carried out on the bacteria liquid blanched, (4) the inactivated vibrio vulnificus is collected by a centrifuge, (5) an ultrasonic crushing is taken on the inactivated vibrio vulnificus collected, (6) immunity stimulated compound ingredients are prepared, and (7) the immunity stimulated compound is prepared by ultrasonic treatment. The virulence of the vibrio vulnificus selected is quite strong (LD50 is about 200cfu / g), the cultivation condition and optimized culture medium can double the biological content of viable bacteria in the fermentation broth, and the biomass of the vibrio vulnificus cultivated is more than 50mg / ml, thus providing energetic technological support for the industrialized fermentation production of the vibrio vulnificus.

Description

technical field [0001] The invention relates to the preparation and application of a pathogenic bacteria inactivated vaccine. Background technique [0002] Vibrio vulnificus (vibrio vulnificus) is a low-grade halophilic Vibrio, which is a zoonosis pathogen in humans and fish, and can cause sepsis and severe wound infection in humans and eels. It can be roughly divided into three types: biological I, biological II, and biological III. Among them, biological type I mainly endangers human health, and can cause a fatality rate of more than 60% after human infection; while biological type II is the main bacterial infection of eels. One of the sources of infectious diseases, in the 1970s and 1980s, the eel industry in Japan, Europe and the United States suffered huge economic losses; in the past five years, the disease caused by Vibrio vulnificus in coastal areas of my country has also been very serious. There are two kinds of processes, acute and protracted, and vibrio vulnificus ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61P31/04C12N1/20
Inventor 许斌福林天龙刘晓东林能锋
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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