| Content of the thesis | 第1-10页 |
| 中文摘要 | 第10-12页 |
| Abstract | 第12-15页 |
| List of abbreviations | 第15-17页 |
| 1. Introduction | 第17-26页 |
| ·Campylobacter jejuni | 第17-18页 |
| ·Campylobacteriosis | 第18-19页 |
| ·Macrolides | 第19-20页 |
| ·Antimicrobial resistance, fitness and virulence in Campylobacter | 第20-22页 |
| ·Aims and objectives of the study | 第22-26页 |
| ·Research contents | 第24-26页 |
| 2. Determination of the cost imposed by different macrolide resistance mutations in Campylobacter jejuni | 第26-47页 |
| ·Introduction | 第26-27页 |
| ·Materials and methods | 第27-35页 |
| ·Bacterial strain and growth conditions | 第27-28页 |
| ·In vitro induction of C.jejuni 1221 | 第28页 |
| ·Antimicrobial susceptibility testing | 第28-29页 |
| ·DNA extraction, primers and PCR reaction conditions | 第29-30页 |
| ·Competition experiments and fitness estimation | 第30页 |
| ·The growth curve of the competent strains | 第30-31页 |
| ·Pairwise competition experiment | 第31-32页 |
| ·Illustrative calculations of selection rate constant (R), Malthusian parameter (M), number of the doubling (D) and relative fitness (W) of each competitor | 第32-33页 |
| ·Heat shock tolerance assay | 第33-34页 |
| ·Colonization of balb/c mice | 第34-35页 |
| ·Statistical analysis | 第35页 |
| ·Results | 第35-44页 |
| ·Determination of mutations in macrolide resistant mutants | 第35-37页 |
| ·Determination of minimal inhibitory concentration (MIC) | 第37-38页 |
| ·Growth rate of S1221 strain compared to STY, SE and STI strains | 第38-39页 |
| ·Relative fitness of S1221 strain compared to STY, SE and STI strains | 第39-41页 |
| ·Heat shock tolerance assay | 第41-42页 |
| ·Colonization of balb/c mice model | 第42-44页 |
| ·Discussion | 第44-47页 |
| 3. Impact of erythromycin resistance on the virulence properties and fitness ofCampylobacter jejuni | 第47-71页 |
| ·Introduction | 第47-49页 |
| ·Materials and methods | 第49-55页 |
| ·Bacterial strain and culture conditions | 第49页 |
| ·In vitro induction | 第49页 |
| ·Antimicrobial susceptibility testing | 第49页 |
| ·C.jejuni genomic DNA extraction and determination of mutations in macrolide action sites | 第49-50页 |
| ·Resistance to bile salts (sodium deoxycholate) | 第50页 |
| ·In vitro cell lines and cells growth conditions | 第50-51页 |
| ·Adhesion and invasion of C.jejuni susceptible strain or Ery-resistant mutant to Caco-2 cell or macrophage RAW 264.7 cells | 第51-52页 |
| ·Intracellular survivability of C.jejuni susceptible strain or Ery-resistant mutant within macrophage RAW 264.7 cells | 第52页 |
| ·Fitness estimation | 第52页 |
| ·Total RNA isolation | 第52页 |
| ·Realtime RT-PCR | 第52-53页 |
| ·Construction of the C.jejuni DNA microarray | 第53-54页 |
| ·Statistical analysis | 第54-55页 |
| ·Results | 第55-68页 |
| ·The mutations in macrolide action sites | 第55页 |
| ·Minimum inhibitory concentration (MIC) | 第55-56页 |
| ·Tolerance to bile salts | 第56-57页 |
| ·Adhesion and invasion characteristics of C.jejuni susceptible strain and Ery-resistant mutant strain to Caco-2 cells | 第57页 |
| ·Adhesion and invasion characteristics of C.jejuni susceptible strain and Ery-resistant mutant strain to macrophage | 第57-58页 |
| ·Survivability of C.jejuni susceptible strain or Ery-resistant mutant strain within the murine macrophage | 第58-60页 |
| ·Fitness analysis of the parent and mutant strains | 第60-61页 |
| ·Total RNA isolation | 第61-62页 |
| ·DNA microarray | 第62-68页 |
| ·Discussion | 第68-71页 |
| 4. Macrolide inhibits flagellar filaments formation without affecting flagellin synthesiswithin Campylobacter jejuni | 第71-88页 |
| ·Introduction | 第71-73页 |
| ·Materials and methods | 第73-77页 |
| ·Bacterial strains and culture conditions | 第73页 |
| ·Determination of bacterial minimum inhibitory concentration | 第73页 |
| ·In vitro induction of C.jejuni 1221 | 第73页 |
| ·Natural transformation of C.jejuni 11168 | 第73-74页 |
| ·Exposure of transformed T strain to sub-inhibitory concentrations of macrolide | 第74页 |
| ·DNA extraction, primers and PCR reaction conditions | 第74-75页 |
| ·Motility test | 第75页 |
| ·Transmission electron microscope | 第75页 |
| ·Purification of secreted protiens and preparation of cell lysate | 第75-76页 |
| ·SDS-PAGE and immunoblotting | 第76-77页 |
| ·Results | 第77-86页 |
| ·Determination of the bacterial minimal inhibitory concentration (MIC) | 第77页 |
| ·The mutations in macrolide action sites | 第77页 |
| ·Characteristics of the transformed T strain | 第77-79页 |
| ·Effect of macrolide on the motility of C.jejuni | 第79页 |
| ·Effect of macrolide on the morphology of C.jejuni | 第79-82页 |
| ·Effect of macrolide on the protein synthesis within C.jejuni | 第82-83页 |
| ·Effect of macrolide on the flagellin synthesis within C.jejuni | 第83-84页 |
| ·Effect of macrolide on the secreted proteins from C.jejuni | 第84-85页 |
| ·Effect of macrolide on flagellin secretion from C.jejuni | 第85-86页 |
| ·Discussion | 第86-88页 |
| 5. General remarks | 第88-94页 |
| ·General discussion | 第88-92页 |
| ·Conclusion | 第92-93页 |
| ·Future work | 第93-94页 |
| 6. The food borne pathogen Campylobacter jejuni:should not remain as an unsung bug | 第94-129页 |
| ·Historical perspective of Campylobacter | 第94-95页 |
| ·Taxonomy | 第95-96页 |
| ·The genome of Campylobacter | 第96-98页 |
| ·Epidemiology of Campylobacter jejuni | 第98-100页 |
| ·Natural transformation in Campylobacter | 第100-102页 |
| ·Antimicrobial resistance in Campylobacter jejuni | 第102-107页 |
| ·Macrolide resistance | 第102-105页 |
| ·Fluoroquinolone Resistance | 第105-106页 |
| ·Tetracycline resistance | 第106-107页 |
| ·Pathogenesis of Campylobacter | 第107-120页 |
| ·Flagella | 第108-111页 |
| ·Chemotaxis | 第111-112页 |
| ·Protein glycosylation | 第112页 |
| ·Lipooligosaccharide and capsular polysaccharide | 第112-113页 |
| ·Adhesion and invasion | 第113-117页 |
| ·Toxins production | 第117页 |
| ·Iron acquisition | 第117-119页 |
| ·Stress reponses | 第119-120页 |
| ·Pathogenesis of diarrhea in C.jejuni infection | 第120-123页 |
| ·Post-infection Complications | 第123-125页 |
| ·Guillain-Barre Syndrome (GBS) | 第124-125页 |
| ·Reiter's syndrome or Reactive Arthritis (ReA) | 第125页 |
| ·Treatment and control of Campylobacter jejuni infection | 第125-129页 |
| 7. Bibliography | 第129-168页 |
| 8. Acknowledgement | 第168-170页 |
| 9. Curriculum Vitae | 第170-172页 |
| 10. Appendix | 第172-186页 |
| ·List of publications | 第172-173页 |
| ·Supplementary materials and methods | 第173-186页 |
