Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

An adjuvanted Riemeria anatipestifer shadow vaccine with chitosan oligosaccharide

A technology of Riemerella anatipestifer and shadow, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of secondary infection, unable to provide mucosal immune barrier against infection, unable to generate local immunity, etc. The effect of preventing viral infection and comprehensive immune protection

Inactive Publication Date: 2018-11-27
QINGDAO AGRI UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since Riemeria anatipestifer has basically lost its ability to colonize the respiratory tract after being prepared as a bacterium, it cannot produce local immunity and can only immunize animals by injection, so there are still problems in the following aspects: ① Immunization by injection, giving Animals cause great immune stress, often causing paramyxovirus and Escherichia coli to take advantage of the weak point, leading to severe secondary infection
② Inactivated vaccines can only induce humoral immunity, no or only slight local mucosal immunity, and cannot provide a reliable mucosal immune barrier against infection

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • An adjuvanted Riemeria anatipestifer shadow vaccine with chitosan oligosaccharide
  • An adjuvanted Riemeria anatipestifer shadow vaccine with chitosan oligosaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Implementation Example 1 Preparation of Riemerella anatipestifer bacterial shadow

[0018] 1.1 Preparation of protoplasts: After activation of the RA2 strain numbered WF1, culture to OD 600 When the value is 0.7, place the bacterial liquid in an ice water bath for 30 minutes, take 10 mL of the bacterial liquid, centrifuge at 4°C, 5000 rpm for 10 minutes, discard the supernatant, wash the bacterial mud with 10 mL of ice-water bath sterilized water, centrifuge again, and repeat the washing three times. Suspend with ice-water bath hypertonic buffer, add preheated lysozyme solution to 5mg / mL, 37°C water bath for 30min, then add preheated EDTA solution to 0.01mol / L, 37°C water bath for 20min. During the action, take the treated bacterial solution on a glass slide every 5 minutes, stain with red, and observe the formation of protoplasts under an oil microscope. When the formation rate of protoplasts reaches 95% or more, centrifuge at 5000 rpm for 20 minutes, discard the supernat...

Embodiment 2

[0022] Implementation example 2 Vaccine immune protection test

[0023] In order to verify the clinical protective effect of the fungus vaccine prepared by the present invention on ducks, commercial meat ducks from Cherry Valley were selected as experimental animals. The fungus vaccine was vaccinated by adding different adjuvants through different immunization routes. At the same time, traditional formaldehyde was used. Control test of oil adjuvant vaccine of homologous strain prepared by inactivation method. The immune protection effect was evaluated by the comparison test of antibody level detection and challenge protection rate.

[0024] 1. Materials and methods

[0025] 1.1 Test materials

[0026] Fungus shadow+chitooligosaccharide adjuvant: prepared according to the method described in Example 1.4.

[0027] Bacteria shadow + oil adjuvant: mix the prepared RA2 strain (WF2) with mineral oil adjuvant in a ratio of 1:1 to make the bacterial count reach 5×10 10 cfμ / mL.

[0028] Formald...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The purpose of the present invention is to provide a riemerella anatipestifer bacterial ghost vaccine loaded with polyinosinic acid-polycytidylic acid having interferon inducing activity and added with chitosan oligosaccharide as an adjuvant. According to the technical scheme, a temperature control expression vector is constructed and is electrotransformed into riemerella anatipestifer protoplast, positive clones are screened and then are subjected to bacterial amplification culture at a temperature of 28 DEG C, lysis gene expression is induced at a temperature of 42 DEG C, polylysine is added to continuously act when the OD600 is no longer be reduced so as to completely lyze the live bacteria, centrifugation is performed to separate the bacteria, the dried bacteria and a 6 mg / mL Poly I:C solution according to an equal ratio so as to make the Poly I:C be embedded into the bacterial ghosts, a 2% chitosan oligosaccharide aqueous solution is added, stirring is performed to form a uniform mixture, sub-packaging is performed into penicillin bottles, and freeze-drying is performed so as to obtain the novel riemerella anatipestifer bacterial ghost vaccine. According to the present invention, the prepared riemerella anatipestifer bacterial ghost vaccine is immunized through drinking water, such that the local immunity on the respiratory tract mucous membrane and the digestive tract mucous membrane can be irritated, the humoral immunity can be stimulated, and the protection effect on the human body is not different from the injection immunity.

Description

Technical field [0001] The invention belongs to the field of veterinary vaccine production, and relates to a preparation method of a bacterial vaccine for ducks, in particular to a preparation method of a Riemerella anatipestifer bacterial shadow vaccine using chitooligosaccharides as an adjuvant. Background technique [0002] Riemerella Anatipestifer (RA) mainly affects ducklings of 2-7 weeks of age. It is characterized by neurological symptoms and fibrinous pericarditis, perihepatitis and air sacculitis. It is currently the main bacterial infectious disease that harms duck production. one. The disease can be infected through respiratory tract, digestive tract and skin wounds, and often forms mixed infections with pathogens such as E. coli and paramyxovirus, causing a large number of ducks to die. [0003] Although Riemerella anatipestifer is sensitive to some antibiotics, it is difficult to completely eliminate it in the body. At the same time, due to the unreasonable use of ant...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/02A61K39/39C12N1/21C12N15/74A61P31/04
Inventor 刘文华刘宗柱任慧英王晓雷张灿
Owner QINGDAO AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com