Preparation Of New Castle-Disease Nanoemulsion

In order to improve the disadvantages of conventional vaccine, we explored New Castle-Disease Nanoemulsion(NDV-NE). Through stability experiment, security experiment, as well as immunogenicity, we provided the basis for New Castle-Disease Nanoemulsion in clinical research and applications.Part 1: The preparation and quality evaluation of New Castle-Disease Nanoemulsion. This new vaccine was prepared by studying the pseudoternary phase diagrams. Tween-80, Span-80, absolute alcohol, ethyl acetate, Liquid paraffin, Levamisole hydrochloride and New Castle-Disease antigen were selected to prepare New Castle-Disease Nanoemulsion. Its structural type, appearance, particle size, viscosity, and physical stability were tested respectively. TEM and Nicomp388/Zeta PALS presented the nanoemulsion as spherical drops with uniform size distribution and mean diameter of 364 nm. The viscosity of compound propolis nanoemulsion was 5s?0.4 mL-1; There was no emulsion breaking in High-speed centrifuge test at 5000r/min in30min; Stability test showed the storage period for 6 moths at 4℃, 6 moths at 20℃, 1 moth at 40℃. New Castle-Disease Nanoemulsion met the requirements of quality inspection of biological products.Part 2: The security evaluation of New Castle-Disease Nanoemulsion. In this section,Through acute toxicity test, Cytotoxicity test and Targeting Animal test to evaluate the safety of New Castle-Disease Nanoemulsion and Nanoemulsion carrier. The results showed that Nanoemulsion carrier belongs to truly nontoxic, LD50>5000 mg/kg. Cytotoxicity test showed that there was no significant toxicity to L929 less than 5μg/mL. It is safe, without vaccine side effects to New Castle-Disease Nanoemulsion by oral or injection. This study provided New Castle-Disease Nanoemulsion is reliable and met safety requirements of biological productsPart 3: The effect of New Castle-Disease Nanoemulsion on antibody titer. In this section, we studied the effects of Levamisole hydrochloride concentration, antigen content and oral immunization dose on antibody titer. The results showed the optimal content of levamisole hydrochloride in NDV-NE was 1%, Newcastle disease antigen was 30%, and the optimal immunization dose was 0.75mL. In this study, we determined the optimal immunization method, and provided the basis for the future research. Part 4: The study on immunization effect of New Castle-Disease Nanoemulsion. Based on the preliminary studies, we studied the effect of New Castle-Disease Nanoemulsion on the immune function through HI test, lymphocyte transformation test, IgA content Determination in local mucosal immune tissues and Challenge Test. The results showed serum HI antibody titers of chickens by oral immunization NDV-NE, IgA levels and lymphocyte transformation rate were significantly increased, which not only induced humoral and cellular immunity, but also stimulated mucosal immunity to play a systemic immune response, forming a broad and effective immune protection. This showed that the nanoemulsion vaccine effect is remarkable.