| Actinobacillu spleuropneumoniae is the primary pathogen of porcine contagious pleuropneumonia. It spreads widely around the world and is one of the infectious diseases which have greatly threatened swine industry.Actinobacillus pleuropneumoniae serotype type 1、3、5、7 which are epidemic in China.According to the situation, there are some number of drug evaluation model,but lack of experimental animal model of infection.So the Kunming mice of SPF level were chosen as experimental animals and the experimental App-S1, App-S7 animal models were established. The classic case of pleuropneumonia was got and drug sensitive tests were taken for the two strains of serotypes.The structure and function of autotransporter adhesions was analyzed by bioinformatics methods.The main research contents and results of this thesis are as follows :1. The establishment of infection model of App-S1 miceAccording to the epidemic bateria App-S1 of China, the Kunming mice of SPF level were chosen as modeling animal, which proved that this bacterial strain has strong pathogenicity for Kunming mice. The LD50 was4.48×105.85 cfu. The infected mice had the typical symptoms of pneumonia and pleurisy with the inflammatory changes in lung, spleen and kidneys and the severe bleeding from the perspective of histopathology. The result showed that the App-S1 is infective and pathogenic to the Kun Ming mice.2. The establishment of infection model of App-S7 miceAccording to the epidemic bacteria App-S7 of china, the Kunming mice of SPF level were chosen as modeling animal to verify the pathogenicity of App-s7 for Kunming mice. The infected mice didn’t have the typical symptoms of pneumonia and pleurisy and there were no clinical features and death. The result showed that App-S7 was the kind of low virulent strain and didn’t have pathogenicity for Kunming mice.3. The Cross-protection test between App-S1 and App-S7Choose virulence strong type 1 and virulence weaker type 7 cross immune protection test research.Firstly,prepare of antigen by App-S7,respectively using live bacteria immunization and inactivated bacteria immunity, immune to mice, after detecting antibodies.With 10 times the LD50 of App- S1 bacteria in mice injection, calculate the protection efficiency,verify whether exists the cross protection between type 1 and type 7 strains.The results showed that the vaccination 7 type of bacteria living bacterium immune antigen and inactivated bacteria immune antigen in mice are of type1 bacteria have immune protection, Recoveries were 80% and 60%respectively, live bacteria immune effectivity is better than that of inactivated bacteria immune.4. The optimization of the App culture and drug susceptibility testBased on the three kinds of mediums for App optimization, such as Chocolate, brain heart immersion, blood tablet mediums.The results showed the medium of Chocolate is the most suitable for App. Drug susceptibility test results show that, in view of the App-S1, medicine norfloxacin,norfloxacin, chloramphenicol, cephalothin Vc are extremely sensitive and complex new Ming and norfloxacin are highly sensitive for the bacteria. In view of the App-S7, Medicine norfloxacin is extremely sensitive and norfloxacin, cnorfloxacin, chloromycetin are highly sensitive.5. Analysis of the protein Adh genomeReported in Genbank App-S5 b from autotransporter adhesion Adh gene DNA sequence alignment and analysis of genetic evolution. At the same time,creatures with the biological software and online database analysis of the basic characteristics of the protein amino acid including physical and chemical properties, hydrophobicity, across the membrane area, curly spiral structure, signal peptide, molecular signaling pathways. The results showed,the protein Adh consists of 3154 amino acids, molecular weight is 316.70 KDa, and the p I is 4.71.No signal peptide, transmembrane area, structure characteristic is given priority to with beta spiral. In protein interactions and intracellular signaling pathway by other interactions of protein such as MKP protein and the Trk protein. |
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