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Oil-adjuvant vaccine of larvae membrane protein of cryptocaryon irritans, preparation method and applications thereof

A technology that stimulates Cryptocaryon and membrane proteins, applied in pharmaceutical formulas, medical preparations containing active ingredients, protozoan antigen components, etc., can solve problems such as economic loss, limited fish toxicity, etc., and achieve high safety, The effect of good vaccine quality and reduced mortality

Inactive Publication Date: 2010-05-12
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, cryptocystosis has been stimulated to break out frequently all over the world, and the damage is becoming more and more serious. "White spot disease" also breaks out in the mariculture areas of the South my country Sea every year, causing great economic losses.
However, the current methods of preventing and treating "white spot disease" in seawater fish are very limited. Although some physical and chemical methods can play a certain role, they all have some defects, such as toxicity to fish, uncertain effect or ineffectiveness. Suitable for large bodies of water, etc.

Method used

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  • Oil-adjuvant vaccine of larvae membrane protein of cryptocaryon irritans, preparation method and applications thereof
  • Oil-adjuvant vaccine of larvae membrane protein of cryptocaryon irritans, preparation method and applications thereof
  • Oil-adjuvant vaccine of larvae membrane protein of cryptocaryon irritans, preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 stimulates the preparation of Cryptocaryon larvae membrane protein oil adjuvant vaccine

[0025] 1. Stimulating Collection of Live Cryptocaryon larvae

[0026] When Cryptocaryon stimuli larvae were used to infect Pomfret ovata, visible trophozoites appeared on the skin and gills of the fish after 60 hours, and almost all the trophozoites were detached from the fish body to form cysts in the next 24 hours, and the detached trophozoites were collected. Cysts, after 60 hours of culture, the cysts began to release larvae. After the larvae hatched, collect the worm fluid, mix well and draw 5 parts (50ul each), add 10ul formalin to kill the larvae, and count them under a microscope. Calculate the concentration of larvae, then add 10% sucrose to the collected worm liquid, mix well, centrifuge at 3500rpm for 10min (KUBOTA 5100 type centrifuge), discard the supernatant, and the precipitate is the collected living larvae.

[0027] 2. Stimulation of Cryptocaryon larv...

Embodiment 2

[0051] The quality inspection of embodiment 2 vaccine

[0052] Get 1 drop of the vaccine prepared in Example 1 and drop it gently in water, observe at rest for 30 minutes and find no oil droplets to scatter; get the prepared vaccine and centrifuge at 3000rmp for 15 minutes without stratification; Coomassie Brilliant Blue R250 (water-soluble dye) and Sudan II ( Oil-soluble dye) detection, the vaccine is the color of Sudan II dye, and the vaccine is a water-in-oil formulation.

Embodiment 3

[0053] The safety detection of embodiment 3 vaccines

[0054] Simultaneously stimulate live Cryptocaryon larvae (2.0×10 4 larvae / tail fish) and pomfret (124.5±38.1g) infected with vaccine soaked with the same number of larvae, all fish infected with live Cryptocaryonia stimuli larvae died within 4 days, and there were a large number of irritants distributed on the body surface and gills. Cryptocaryon trophozoites, but the fish soaked by the stimulated Cryptocaryonia vaccine did not appear to die, and the distribution of Cryptocaryon trophozoites was not found on the body surface and gills, which proved that the vaccine was safe to use in production .

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Abstract

The invention discloses an oil-adjuvant vaccine of larvae membrane protein of cryptocaryon irritans, and a preparation method and applications thereof. The preparation method of the vaccine comprises the following steps: (1) collecting live larvae of the cryptocaryon irritans; (2) extracting the larvae membrane protein of the cryptocaryon irritans; and (3) adding PBS in the larvae membrane protein as water phase and mixing No.10 white oil, Span-80 and aluminum stearate as oil phase, emulsifying the larvae membrane protein in the volume ratio of the water phase to the oil phase, which is 3:7, thus obtaining the oil-adjuvant vaccine of the larvae membrane protein of the cryptocaryon irritans. The preparation method of the oil-adjuvant vaccine of the larvae membrane protein of the cryptocaryon irritant is simple and practicable, and the obtained oil-adjuvant vaccine of the larvae membrane protein of the cryptocaryon irritans has high safety, obvious immunoprotection effect and no toxicity to fish, can be used for preventing and treating the parasitic infection of sea fish and can absolutely meet the demand of actual production.

Description

technical field [0001] The invention relates to the field of vaccine preparation, in particular to a membrane protein oil adjuvant vaccine that stimulates Cryptocaryon larvae and its preparation method and application. Background technique [0002] Cryptocaryon irritans Brown (1951), also known as Ichthyophthirius marinus, is an obligate fish parasite that parasitizes in tropical and subtropical seawater fish and causes the host to suffer from "white spot disease". ". The host range of C. stimuli is very wide. Except for cartilaginous fishes which have strong resistance to C. stimuli, bony fishes in seawater can generally be infected by C. stimuli. Severe infection will lead to a large number of fish deaths. In recent years, cryptocystosis has frequently erupted all over the world, and the damage is becoming more and more serious. The "white spot disease" also breaks out in the mariculture areas of the South my country Sea every year, causing great economic losses. However...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/002A61K39/39A61P33/02
Inventor 李安兴但学明柏建山王芳华
Owner SUN YAT SEN UNIV
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