An adjuvanted Riemeria anatipestifer shadow vaccine with chitosan oligosaccharide

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...

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

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