Immune-enhancing Effects Of The Mixtures Of Taishan Pinus Massoniana Pollen Polysaccharide, Aloe Polysaccharide And Proplis On Immunized Chickens

Animal infectious diseases caused by bacterial and viral pathogens are increasingly complex and severe. Drugs and antibiotics play important roles in controlling these diseases, but the frequent use of drugs generally leaves residues and leads to the emergence of drug-resistant strains. Vaccination is the most effective method to prevent the emergence of the infectious diseases. Generally, immunologic adjuvants are applied in the preparation of vaccines, because they can enhance the immune response of vaccines and extend the duration of immunity-enhancing effects.Our previous study revealed that Taishan Pinus massoniana pollen polysaccharide, aloe polysaccharide and propolis were proved to have effective immune regulation.However, we are not clear whether the effects of these compounds are synergistic ? We used the best dose of Taishan Pinus massoniana pollen polysaccharide, propolis and aloe polysaccharide, investigated whether the combined adjuvants of plant polysaccharides and propolis could induce better effects on chickens inoculated with B.avium inactivated vaccine, ND attenuated vaccine immuning chickens as a breakthrough point, by measuring the humoral and cellular immunity related immune index, to explore different immune immunomodulators and their mixtures on the immune enhancement effect in chickens, select the best formula, for livestock exposed a new generation of vaccine adjuvant or immune enhancer provides theory basis for research and development.This research is divided into three aspects:1. Extraction of polysaccharides and their contents measureUsing the improved Water extract-alcohol method to extract Taishan Pinus massoniana pollen polysaccharide, aloe polysaccharide, and through the phenol-sulfuric acid method to determine the polysaccharide extraction respectively.This method under the Taishan Pinus massoniana pollen polysaccharide, aloe polysaccharide yield were 4.93%, 2.67%, polysaccharide content was 76.4%, 80.4%.2. Study on immune enhance effect of different immune adjuvants and their mixtures on B. avium inactivated vaccines.To study immune enhancement effects of the combined use of Taishan Pinus massoniana pollen polysaccharide, aloe polysaccharide and proplis on Bordetella avium inactivated vaccine. 160 3 days age SPF chickens were randomly divided into 8groups. At the age of 3, 7, and 12, In TPPPS, PP and AP group chickens were were inoculated subcutaneously with TPPPS, PP, and AP, respectively; In TPPPS-PP, PP-AP,TPPPS-AP group chickens were inoculated subcutaneously with TPPPS-PP, PP-AP,and TPPPS-AP, respectively; PBS group chickens were subcutaneously inoculated with B. avium inactivated vaccines, which contained an equal amount of PBS.Chickens in group Mock were only inoculated with equivalent physiological saline. At7, 14, 21, 28, 35, and 42 dpi, We take three chickens blood samples for each group randomly and detect serum antibodies, the blood content of IL-2, the lymphocyte transformation rates and peripheral blood CD4+, CD8+T-lymphocytes count,respectively. The results showed that all the immune indicators of group TPPPS, PP,TPPPS-PP at 14, 21, 28, 35 dpi were higher than that of the other groups, and were significantly higher than that of PBS, Mock group( P < 0.05), at 14, 21,28 dpi the group TPPPS-PP had the highest level(P < 0.05). The results revealed that Taishan Pinus massoniana pollen polysaccharide and propolis cound induce the highest level of humoral and cellular immune response. Therefore, There were the synergistic effect between Taishan Pinus massoniana pollen polysaccharide and propolis on enhancing immunity effect of B. avium inactivated vaccine.3. Study on immune enhance effect of different immune enhancer and their mixtures on NDV vaccines.To study immune enhancement effects of the combined use of Taishan Pinus massoniana pollen polysaccharide, aloe polysaccharide and proplis on ND attenuated vaccine. 160 3 days age SPF chickens were randomly divided into 8 groups. At the age of 12, NDV attenuated vaccines firstly were inoculated with eye and nasal dropping,and at the age of 28 NDV attenuated vaccines secondly were inoculated subcutaneously. In TPPPS, PP and AP group chickens were inoculated subcutaneously TPPPS, PP, and AP, respectively; In TPPPS-PP, PP- AP, TPPPS-AP group chickens inoculated subcutaneously with TPPPS-PP, PP-AP, TPPPS-AP; In PBS group chickenswere inoculated subcutaneously amount of PBS. In MOCK group, chickens were only inoculated subcutaneously with amount of saline solution. At 7, 14, 21, 28, 35, and 42 dpi, We take three chickens blood samples for each group randomly and detect serum antibodies, the blood content of IFN-γ, the lymphocyte transformation rates and peripheral blood CD4+, CD8+T-lymphocytes count, respectively. The results showed that all the immune indicators of 7 vaccine groups at 14,21,28,35,42 dpi were higher than that of MOCK group, at 14,21,28 dpi that of 6 immunoenhancement groups were significantly higher than that of PBS group( P < 0.05), and at peak value dpi the group TPPPS-PP had the highest level(P < 0.05). The results revealed that Taishan Pinus massoniana pollen polysaccharide and propolis cound induce the highest level of humoral and cellular immune response. Therefore, There were the synergistic effect between Taishan Pinus massoniana pollen polysaccharide and propolis on enhancing immunity effect of ND attenuated vaccine.