Studies On The Extraction And Separation Of Polysaccharides From Sargassum And Its Effect For Immune Response To PRRS Virus As Immunity Adjuvant

This paper has explored the method of extraction and separation and purition of polysaccharides from Sargassum , the specific Sargassum species in WeiZhouDao, BeiHai, GuangXi, and determinated its molecular weight and components. The polysaccharides was examined for the effect of immune response as adjuvant.Part 1. Studies on the extraction and separation and purification of polysaccharides from Sargassum.It took twenty minutes to pretreat the Sargassum with ultrasonic , in order to break the cell wall, then extracting 3h at 80℃, filtrating, certrifuging .condensing, depositing the sulphated polysaccharides with alohol, and drying . The extraction rate of polysaccharides was 13. 12%, and increased respectively by 8.73% and 1.39% comparing to traditional method and Colloid milling, and decreased by 300 minuts and 420 minuts for extraction time. Then the polysaccharides SP was made by filtrating and enzymolysis.By grading the Sargassum polysaccharides SP over DEAE-cellulosecolume and through separation , we found that the polysaccharides included three components (SP-I, SP-II, SP-III), the percentage were 31.4%, 23.8% and 8.1% respectively. Then further separation and purification steps that were done through chromato graphyon SephadexG-100 column were used to obtain a fraction named SSP-I, SSP-II, SSP-III. Them molecular weight were 53862,46536 and 49866 respectively by the same way .Theirchimical ingredients and rations shown as follows, SSP-I: Ara: Xyl: Glu: Gal =0.012: 0.18: 0.676: 0. 144;SSP-II: Ara: Xyl: Glu: Gal -0.015: 0. 178:0.685 :0. 122; SSP-III: Rha: Xyl : Glu:Ara: Gal =0.039: 0. 121:0.726:0. 114. Part 2. The effect for immune response to PRRS virus vaccine asadjuvantExperimental groups were vaccimated with normal adjuvant vaccine and three kinds of complete adjuvant vaccines which used 50mg, lOOmg, 200mg polysaccharides as adjuvant respectively +inactivity PRRS antigen; every group have five non-immune pigs. Blooding and detemining its cell-immunity and humoral-immunity standard in the ON 20> 40 ., 50 day after primary vaccinate.The results showed (1)T lymphocyte transformation rate of polysaccharides groups are :61.4%, 65%, 63.4% all higher than control group 52.8% and normal adjuvant group 59.6%. And there was a obviouse defference betwen polysaccharides groups and control group(P<0. 05);(2) Qutities of T lymphocyte subsets were all higher than the other groups,the increasion of CD3 was the most obvious. But comparing of CD8 and NK cell was not significant;(3)Antibody obsordance in serum from polysaccharides groups were 0.631,1.015,0.688 .which was higher than control group 0. 518 and normal group 0. 568 determinated by the method of indirectly ELISA, and the defference was significant (P<0. 05).The studies of result demonstrated that Sargassumpolysaccharides as adjuvant could improve humoral- immunity and enhance cell-immunity. Among the three polysaccharides groups, the total result of lOOmg group was the best.