Jin Yongjie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 138-171). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iii / Acknowledgments --- p.iv / Table of Contents --- p.v / List of Tables --- p.x / List of Figures --- p.xi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1. --- Salmonella --- p.1 / Chapter 1.1 --- Morphology --- p.1 / Chapter 1.2 --- Antigenic structure --- p.1 / Chapter 1.3 --- Identification --- p.2 / Chapter 1.4 --- Nomenclature and classification --- p.2 / Chapter 1.5 --- Pathogenesis and virulence --- p.3 / Chapter 1.6 --- Infections --- p.4 / Chapter 1.6.1 --- Enteric fever --- p.4 / Chapter 1.6.2 --- Gastroenteritis --- p.4 / Chapter 1.7 --- Treatment --- p.5 / Chapter 1.7.1 --- Enteric fever --- p.5 / Chapter 1.7.2 --- Gastroenteritis --- p.5 / Chapter 1.8 --- Epidemiology and control --- p.6 / Chapter 1.8.1 --- Enteric fever --- p.6 / Chapter 1.8.2 --- Salmonella food poisoning --- p.6 / Chapter 2. --- Fluoroquinolones --- p.6 / Chapter 2.1 --- History of fluoroquinolones --- p.8 / Chapter 2.2 --- Mechanisms of action --- p.9 / Chapter 3. --- Antimicrobial resistance --- p.10 / Chapter 3.1 --- Microorganism-mediated resistance --- p.10 / Chapter 3.1.1 --- Intrinsic resistance --- p.11 / Chapter 3.1.2 --- Acquired resistance --- p.11 / Chapter 3.2 --- Resistance due to environmental factors --- p.12 / Chapter 3.3 --- Biological and clinical resistance --- p.12 / Chapter 3.4 --- Breakpoints --- p.13 / Chapter 4. --- Fluoroquinolone-resistance --- p.14 / Chapter 4.1 --- Increasing fluoroquinolone-resistance in bacteria --- p.14 / Chapter 4.2 --- Mechanisms of resistance to fluoroquinolones --- p.16 / Chapter 4.2.1 --- Mutations of target genes --- p.17 / Chapter 4.2.2 --- Active efflux systems and decreased membrane permeability --- p.20 / Chapter 5. --- Fluoroquinolone-resistance in Salmonella --- p.21 / Chapter 5.1 --- Prevalence of fluoroquinolone-resistant salmonellae in man and Animals --- p.21 / Chapter 5.1.1 --- Prevalence in the world --- p.21 / Chapter 5.1.2 --- Increasing resistance trend in Hong Kong --- p.24 / Chapter 5.2 --- Clinical outcome --- p.25 / Chapter 5.3 --- Mechanisms of fluoroquinolone-resistance in Salmonella --- p.25 / Chapter 5.3.1 --- Target gene mutations --- p.25 / Chapter 5.3.2 --- Other resistance mechanisms --- p.28 / Chapter 5.3.3 --- In vitro development of Salmonella resistant mutants --- p.29 / Chapter 6. --- Restricting the development of resistant mutants --- p.31 / Chapter 6.1 --- Mutant prevention concentration (MPC) --- p.31 / Chapter 6.1.1 --- Definition --- p.31 / Chapter 6.1.2 --- Development of MPC concept --- p.31 / Chapter 6.1.3 --- Significance --- p.34 / Chapter 6.2 --- Mutant selection window (MSW) --- p.35 / Chapter 6.2.1 --- The concept of mutant selection window (MSW) --- p.35 / Chapter 6.2.2 --- Significance --- p.36 / Chapter 7. --- Detection of gene mutations --- p.37 / Chapter 8. --- Objectives --- p.37 / Chapter Chapter 2 --- Materails and Methods --- p.39 / Chapter 1. --- Materials --- p.39 / Chapter 1.1 --- Bacterial strains --- p.39 / Chapter 1.1.1 --- Bacterial strains used for this study --- p.39 / Chapter 1.1.2 --- Storage of bacterial strains --- p.39 / Chapter 1.2 --- Materials --- p.40 / Chapter 2. --- Methods --- p.40 / Chapter 2.1 --- Identification --- p.40 / Chapter 2.2 --- Microbiological methods --- p.40 / Chapter 2.2.1 --- Determination of minimal inhibitory concentration (MIC) --- p.40 / Chapter 2.2.1.1 --- Preparation of antibiotic plates --- p.40 / Chapter 2.2.1.2 --- Preparation of inoculum --- p.43 / Chapter 2.2.1.3 --- Inoculation of antibiotic plates --- p.43 / Chapter 2.2.2 --- Determination of mutant prevention concentration (MPC) --- p.43 / Chapter 2.2.2.1 --- Preparation of bacterial suspension --- p.43 / Chapter 2.2.2.2 --- Inoculation of bacterial suspension --- p.43 / Chapter 2.2.2.3 --- Determination of the size of the inoculum --- p.43 / Chapter 2.2.2.4 --- Reading of results --- p.45 / Chapter 2.3 --- Molecular methods --- p.45 / Chapter 2.3.1 --- Polymerase chain reaction (PCR) --- p.45 / Chapter 2.3.2 --- Agarose gel electrophoresis --- p.47 / Chapter 2.3.3 --- Multiplex PCR amplimer conformation (MPAC) analysis --- p.47 / Chapter 2.3.3.1 --- Preparation of samples --- p.47 / Chapter 2.3.3.2 --- Electrophoresis --- p.49 / Chapter 2.3.3.3 --- Silver staining --- p.49 / Chapter 2.3.4 --- DNA Sequencing --- p.50 / Chapter 2.3.4.1 --- Purification of PCR products --- p.50 / Chapter 2.3.4.2 --- Sequencing reactions --- p.50 / Chapter 2.3.4.3 --- Electrophoresis --- p.50 / Chapter 2.3.4.4 --- Silver staining --- p.52 / Chapter 2.4 --- Stepwise selection and characterization of mutants --- p.53 / Chapter 2.4.1 --- In vitro selection of first-step strains --- p.53 / Chapter 2.4.2 --- Characterization of selected strains --- p.53 / Chapter 2.4.3 --- Subsequent selection of stepwise mutants --- p.54 / Chapter 3. --- Research plan --- p.54 / Chapter Chapter 3 --- Results --- p.55 / Chapter 1. --- Identification and fluoroquinolone MICs of Salmonella strains --- p.55 / Chapter 1.1 --- Identification of Salmonella strains --- p.55 / Chapter 1.2 --- Fluoroquinolone MICs for Salmonella strains --- p.55 / Chapter 1.2.1 --- Salmonella Typhimurium --- p.55 / Chapter 1.2.2 --- Salmonella Hadar --- p.57 / Chapter 2. --- Mutant prevention concentration (MPC) --- p.57 / Chapter 2.1 --- Salmonella Typhimurium --- p.57 / Chapter 2.1.1 --- MPC value --- p.57 / Chapter 2.1.2 --- MPC/MIC ratio --- p.57 / Chapter 2.1.3 --- Cmax/MPC ratio --- p.60 / Chapter 2.1.4 --- Frequencies of development of resistant strains --- p.60 / Chapter 2.2 --- Salmonella Hadar --- p.63 / Chapter 2.2.1 --- MPC value --- p.63 / Chapter 2.2.2 --- MPC/MIC ratio --- p.63 / Chapter 2.2.3 --- Cmax/MPC ratio --- p.63 / Chapter 2.2.4 --- Frequencies of development of resistant strains --- p.65 / Chapter 3. --- Stepwise selection of resistant mutants --- p.65 / Chapter 3.1 --- Salmonella Typhimurium --- p.70 / Chapter 3.1.1 --- Characterization of 1 St-step strains --- p.70 / Chapter 3.1.1.1 --- Antimicrobial susceptibilities --- p.70 / Chapter 3.1.1.2 --- Characterization of gene mutations --- p.75 / Chapter 3.1.1.3 --- Mutations and fluoroquinolone susceptibilities --- p.80 / Chapter 3.1.1.4 --- Mutations and nalidixic acid susceptibilities --- p.81 / Chapter 3.1.2 --- Characterization of 2nd-step mutants --- p.81 / Chapter 3.1.2.1 --- Antimicrobial susceptibilities --- p.82 / Chapter 3.1.2.2 --- Characterization of gene mutations --- p.85 / Chapter 3.1.2.3 --- Mutations and fluoroquinolone susceptibilities --- p.87 / Chapter 3.1.2.4 --- Nalidixic acid susceptibilities --- p.87 / Chapter 3.1.3 --- Characterization of 3rd-step mutants --- p.88 / Chapter 3.1.3.1 --- Antimicrobial susceptibilities --- p.88 / Chapter 3.1.3.2 --- Characterization of gene mutations --- p.92 / Chapter 3.1.3.3 --- Mutations and fluoroquinolone susceptibilities --- p.94 / Chapter 3.1.3.4 --- Nalidixic acid susceptibilities --- p.95 / Chapter 3.1.4 --- Characterization of 4th-step mutants --- p.95 / Chapter 3.1.4.1 --- Antimicrobial susceptibilities --- p.95 / Chapter 3.1.4.2 --- Characterization of gene mutations --- p.98 / Chapter 3.1.4.3 --- Nalidixic acid susceptibilities --- p.98 / Chapter 3.2 --- Salmonella Hadar --- p.98 / Chapter 3.2.1 --- Characterization of 1 St-step strains --- p.98 / Chapter 3.2.1.1 --- Antimicrobial susceptibilities --- p.99 / Chapter 3.2.1.2 --- Characterization of gene mutations --- p.102 / Chapter 3.2.1.3 --- Mutations and fluoroquinolone susceptibilities --- p.107 / Chapter 3.2.1.4 --- Mutations and nalidixic acid susceptibilities --- p.108 / Chapter 3.2.2 --- Characterization of 2nd-step mutants --- p.109 / Chapter 3.2.2.1 --- Antimicrobial susceptibilities --- p.109 / Chapter 3.2.2.2 --- Characterization of gene mutations --- p.112 / Chapter 3.2.2.3 --- Mutations and fluoroquinolone susceptibilities --- p.112 / Chapter 3.2.2.4 --- Nalidixic acid susceptibilities --- p.113 / Chapter Chapter 4 --- Discussion --- p.114 / Chapter 1. --- Susceptibility of salmonellae to fluoroquinolones --- p.114 / Chapter 2. --- The potential of fluoroquinolones to restrict the development of Salmonella resistant strains --- p.114 / Chapter 2.1 --- MPC and MPC/MIC of fluoroquinolones --- p.115 / Chapter 2.2 --- Cmax/MPC of fluoroquinolones --- p.117 / Chapter 2.3 --- Selection frequency of fluoroquinolones --- p.118 / Chapter 2.4 --- Effects of fluoroquinolones on the development of resistancein Salmonella Typhimurium and Salmonella Hadar --- p.119 / Chapter 3. --- Characterization of in vitro fluoroquinolone-resistant Salmonella mutants --- p.120 / Chapter 3.1 --- Development of resistance phenotype --- p.120 / Chapter 3.1.1 --- Microbiology --- p.120 / Chapter 3.1.2 --- Antimicrobial susceptibilities --- p.120 / Chapter 3.1.2.1 --- First-step strains --- p.120 / Chapter 3.1.2.2 --- Second-step mutants --- p.121 / Chapter 3.1.2.3 --- Third-step mutants --- p.121 / Chapter 3.1.2.4 --- Fourth-step mutants --- p.122 / Chapter 3.2 --- Contribution of target gene mutations to resistance development --- p.122 / Chapter 3.2.1 --- First-step mutants --- p.122 / Chapter 3.2.2 --- Second-step mutants --- p.125 / Chapter 3.2.3 --- Third-step mutants --- p.126 / Chapter 3.2.4 --- Fourth-step mutants --- p.128 / Chapter 3.3 --- The sequential development of gene mutations --- p.129 / Chapter 3.4 --- Other fluoroquinolone-resistance mechanisms --- p.130 / Chapter 4. --- Mutations and susceptibilities to fluoroquinolones and nalidixic acid --- p.132 / Chapter 4.1 --- Nalidixic acid - a marker for resistance to fluoroquinolones --- p.132 / Chapter 4.2 --- Breakpoint and clinical efficiency --- p.133 / Chapter 5. --- Strategies to reduce development of fluoroquinolone-resistance --- p.134 / Chapter 6. --- Conclusion --- p.136 / Chapter 7. --- Areas for future research --- p.136 / References --- p.138
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324634 |
| Date | January 2004 |
| Contributors | Jin, Yongjie., Chinese University of Hong Kong Graduate School. Division of Microbiology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xi, 171 leaves : ill. ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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