Study Of Echinococcus Granulosus And The Compound Of Praziquantel-attenuated Rabies Vaccine

Echinococcosis granulosus, which is also called cystic echinococcosis, is a zoonotic parasitic disease of worldwide distribution. Inner Mongolia Autonomous Region is the focus areas of national hydatid disease prevention and control, but the effect is unsatisfactory, for the understanding and treatment of this disease has not been paid the attention to which it deserved. In this study, epidemiological investigations on Echinococcus granulosus and its main host was conducted in Inner Monglia, and the phylogenetic analysis was done to CO1 and ND1 gene of the Inner Mongolia strains. At the same time, complex formulation of E.granulosus repellent praziquantel and attenuated rabies vaccine was developed. The results were as follows:1. The antigen detection of Echinococcus granulosus in final host-dogs showed that, high incidence of Echinococcus granulosus were in Wulatezhongqi, Dongwuqi, Xiwuqi, Siziwangqi, and Xilinhot, and the positive rate of samples were 20%,13.8%,13.3%,8% and 3.8%, respectively.2. The infection results of intermediate host of sheep, cattle, and human indicated that adult sheep had low infection with Echinococcus granulosus, while young sheep and bovine were not infected by E.granulosus. Hydatids removed from six patients were proved to be Echinococcus granulosus by parasitology test.3. After being sequenced, the CO1 sequence length of human strains and sheep strains in Inner Mongolia were 936bp, and homology analysis indicated there was high homology between sheep strains and Mongolia strains, while the human strains had high homology with Iran strains. Similarities were both more than 99%, and the two strains in Inner Mongolia were all G1-type in genotype.4. The sequence length of ND1 gene of human strains and sheep strains in Inner Mongolia were all 895bp, and sequence of sheep strains and human strains were same. Similarities of ND1 of the Inner Mongolia strains and Iran strains were 100%, both of them were also G1-type genotype.5. Development process of mixtures of deworming medicine and vaccine displayed that, praziquantel and rabies attenuated vaccine were relatively stable in solution containing 1% Tween-80 ethyl lactate-isopropanol (ethyl lactate:isopropanol=8:2).6. Praziquantel content in praziquantel solution, and mixtures of praziquantel and rabies attenuated vaccine was determined by HPLC.The content was more than 90%, and it met the requirements of the drug content. Then the praziquantel solution and the mixtures were stored in preservation environment of the rabies vaccine. The results showed:no precipitate was saw in the two solutions, and drug content, pH, clarity could meet requirements.7. The abnormal toxicity experimental results of praziquantel- rabies attenuated vaccine compound in mice showed that:mice were in good health and no death came out. The local stimulation test also showed that there was no phenomenon of swelling and hyperaemia in injection site.8. Pre-treatment methods of determining praziquantel in rabbit plasm were screened by high performance liquid chromatography. The results showed that:acetonitrile was the ideal praziquantel extraction materia from the rabbit plasma. Then the HPLC method of determining opraziquantel by gradient elution praziquantel with acetonitrile and water was established.9. Pharmacokinetic studies of the mixtures of praziquantel-attenuated rabies vaccine presented a similar situation of praziquantel administrated only in test animals, and it increased rapidly at 15min, and up to peak at 1h, then it reached the low at 4h. From then on, metabolism was slow and only trace praziquantel was detected at 24h.