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Chio Weng Fan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 131-149). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / Contents --- p.iv / List of tables --- p.ix / List of figures --- p.x / List of abbreviation --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Biology of Staphylococci --- p.1 / Chapter 1.1.1 --- Taxonomy --- p.1 / Chapter 1.1.2 --- Characteristics of S. aureus --- p.2 / Chapter 1.2 --- Methicillin-resistant Staphylococcus aureus --- p.1 / Chapter 1.2.1 --- Description of MRSA --- p.7 / Chapter 1.2.2 --- Antibiotic Resistance of MRSA --- p.7 / Chapter 1.2.2.1 --- Resistance to β-lactam Antibiotics --- p.8 / Chapter 1.2.2.1.1 --- Production of β-lactamase --- p.8 / Chapter 1.2.2.1.2 --- Penicillin-binding Protein PBP2a --- p.8 / Chapter 1.2.2.1.3 --- Borderline Methicillin Resistance --- p.11 / Chapter 1.2.2.2 --- Resistance to Antibiotics other than β-lactams --- p.11 / Chapter 1.2.3 --- Epidemiology of MRSA --- p.15 / Chapter 1.2.3.1 --- Clinical Significance of MRSA --- p.15 / Chapter 1.2.3.1.1 --- Evolution of MRSA --- p.16 / Chapter 1.2.3.1.2 --- Epidemiology of MRSA Worldwide --- p.17 / Chapter 1.2.3.2 --- MRSA in Hong Kong --- p.21 / Chapter 1.3 --- Strain Typing for MRSA --- p.26 / Chapter 1.3.1 --- Phenotypic Methods --- p.27 / Chapter 1.3.1.1 --- Antibiotic Susceptibility Test --- p.27 / Chapter 1.3.1.2 --- Bacteriophage Typing --- p.28 / Chapter 1.3.1.3 --- Multilocus Enzyme Electrophoresis --- p.29 / Chapter 1.3.2 --- Genotypic Methods --- p.30 / Chapter 1.3.2.1 --- Analysis of Chromosomal DNA --- p.30 / Chapter 1.3.2.1.1 --- Pulsed-field Gel Electrophoresis (PFGE) --- p.30 / Chapter 1.3.2.1.2 --- Amplified Fragment Length Polymorphism (AFLP) --- p.31 / Chapter 1.3.2.2 --- Analysis of Gene Polymorphism --- p.32 / Chapter 1.3.2.2.1 --- PCR-Restriction Length Polymorphism (PCR-RFLP) --- p.32 / Chapter 1.3.2.2.2 --- Random Amplification of Polymorphic DNA (RAPD) --- p.33 / Chapter 1.3.2.2.3 --- Nucleotide Sequence Typing --- p.33 / Chapter 1.3.2.2.3.1 --- A Typing --- p.33 / Chapter 1.3.2.2.3.2 --- Multilocus Sequence Typing --- p.33 / Chapter 1.3.2.3 --- Hybridization by Southern Blotting --- p.34 / Chapter 1.3.2.3.1 --- MecA/Tn544 Probe Typing --- p.34 / Chapter 1.3.2.3.2 --- Binary Typing --- p.34 / Chapter 1.3.2.4 --- Analysis of Plasmid DNA --- p.35 / Chapter 1.4 --- Objectives of the Project --- p.37 / Chapter Chapter 2 --- Materials & Methods --- p.38 / Chapter 2.1 --- Bacterial Isolates & Culture Conditions --- p.38 / Chapter 2.1.1 --- Study Period & Sample Sources --- p.38 / Chapter 2.1.2 --- Selection of Bacterial Isolates --- p.38 / Chapter 2.1.3 --- Reference Strains --- p.38 / Chapter 2.1.4 --- Culture & Storage Conditions --- p.39 / Chapter 2.1.5 --- Identification of MRSA --- p.39 / Chapter 2.2 --- Antibiotic Susceptibility Testing --- p.40 / Chapter 2.3 --- PCR for MecA Gene --- p.43 / Chapter 2.3.1 --- DNA Preparation and Primer Design for mecA Gene --- p.43 / Chapter 2.3.2 --- PCR Amplification of mecA Gene --- p.45 / Chapter 2.3.2.1 --- Master Mix Preparation --- p.45 / Chapter 2.3.2.2 --- Polymerase Chain Reaction --- p.45 / Chapter 2.3.2.3 --- Analysis of PCR Products by Agarose Gel Electrophoresis --- p.45 / Chapter 2.3.2.4 --- Precautions to Avoid Cross-contamination --- p.46 / Chapter 2.4 --- Pulsed-field Gel Electrophoresis (PFGE) --- p.47 / Chapter 2.4.1 --- PFGE Protocol --- p.47 / Chapter 2.4.1.1 --- DNA Preparation for PFGE --- p.47 / Chapter 2.4.1.2 --- Restriction Enzyme Digestion --- p.48 / Chapter 2.4.1.3 --- Pulsed-field Gel Electrophoresis --- p.48 / Chapter 2.4.1.4 --- Standards & Markers for PFGE --- p.48 / Chapter 2.4.2 --- Optimization of PFGE --- p.49 / Chapter 2.4.3 --- Computer Analysis of PFGE Patterns --- p.49 / Chapter 2.5 --- Amplified Fragment Length Polymorphism (AFLP) --- p.52 / Chapter 2.5.1 --- Amplified Fragment Length Polymorphism Protocol --- p.52 / Chapter 2.5.1.1 --- DNA Preparation --- p.52 / Chapter 2.5.1.2 --- Enzyme Digestion & Ligation --- p.53 / Chapter 2.5.1.3 --- PCR for AFLP --- p.53 / Chapter 2.5.1.4 --- Gel Electrophoresis for AFLP --- p.54 / Chapter 2.5.1.5 --- Standards & Markers for AFLP --- p.55 / Chapter 2.5.2 --- Computer Analysis for AFLP Patterns --- p.55 / Chapter 2.6 --- Phage Typing --- p.58 / Chapter 2.6.1 --- Phages & Propagating Strains --- p.58 / Chapter 2.6.2 --- Phage Typing Protocol --- p.58 / Chapter 2.6.3 --- Data Analysis and Results Interpretation for Phage Tying --- p.62 / Chapter 2.7 --- Other Typing Methods --- p.63 / Chapter 2.7.1 --- PCR Restriction Fragment Length Polymorphism for the coa gene --- p.63 / Chapter 2.7.1.1 --- Primer Design --- p.63 / Chapter 2.7.1.2 --- DNA Preparation --- p.63 / Chapter 2.7.1.3 --- Optimization of PCR --- p.63 / Chapter 2.7.1.3.1 --- PCR Amplification --- p.64 / Chapter 2.7.1.3.2 --- Restriction Enzyme Digestion --- p.64 / Chapter 2.7.2 --- Multilocus Sequence Typing --- p.64 / Chapter 2.7.2.1 --- Preparation of Chromosomal DNA --- p.65 / Chapter 2.7.2.2 --- PCR for MLST Gene --- p.65 / Chapter 2.7.2.3 --- Purification of DNA for Sequencing --- p.67 / Chapter 2.7.2.4 --- PCR for Sequencing --- p.67 / Chapter 2.7.2.5 --- Sequencing by Automatic DNA Analyzer & Data Analysis --- p.68 / Chapter Chapter 3 --- Results --- p.69 / Chapter 3.1 --- Bacterial Isolates --- p.69 / Chapter 3.2 --- Antibiotic Susceptibility Testing --- p.72 / Chapter 3.3 --- PCR for MecA Gene --- p.78 / Chapter 3.4 --- Pulsed-field Gel Electrophoresis --- p.80 / Chapter 3.4.1 --- Optimization of PFGE --- p.80 / Chapter 3.4.2 --- Analysis of PFGE Profiles --- p.83 / Chapter 3.5 --- Amplified Fragment Length Polymorphism --- p.95 / Chapter 3.6 --- Phage Typing --- p.102 / Chapter 3.7 --- Other Typing Methods --- p.109 / Chapter 3.7.1 --- PCR Restriction Fragment Length Polymorphism for the Coa Gene --- p.109 / Chapter 3.7.1.1 --- Optimization of PCR conditions for the Coa Gene --- p.109 / Chapter 3.7.1.2 --- Analysis of MRSA by PCR-RFLP of the coa Gene --- p.109 / Chapter 3.7.2 --- Multilocus Sequence Typing --- p.115 / Chapter Chapter 4 --- Discussion --- p.116 / Chapter 4.1 --- Evaluation of Typing methods for MRSA --- p.116 / Chapter 4.1.1 --- Antibiotic Susceptibility Testing --- p.116 / Chapter 4.1.2 --- Phage Typing --- p.117 / Chapter 4.1.3 --- Pulsed-field Gel Electrophoresis --- p.118 / Chapter 4.1.4 --- Amplified Fragment Length Polymorphism --- p.119 / Chapter 4.1.5 --- PCR-Restriction Fragment Length Polymorphism for the Coa Gene --- p.120 / Chapter 4.1.6 --- Multilocus Sequence Typing --- p.121 / Chapter 4.1.7 --- Conclusion for Method Evaluation --- p.122 / Chapter 4.2 --- "Analysis of Results of Antibiotic Susceptibility Test, Phage Typing, PFGE, AFLP, PCR-RFLP (Coa) & MLST" --- p.124 / Chapter 4.2.1 --- Correlation between Methods --- p.124 / Chapter 4.2.2 --- Clinical Correlation --- p.125 / Chapter 4.2.3 --- Correlation between MRSA Isolates at PWH and Reference Strains --- p.128 / Chapter 4.3 --- Future Research --- p.129 / Chapter 4.4 --- Conclusion --- p.130 / Reference List --- p.131 / Appendix I Reagent & Material Lists for Methods --- p.150 / Appendix II Buffers & Media --- p.156 / Appendix III Coa Gene Sequences of Isolates from PWH and Reference Strains --- p.158 / Appendix IV Unique antibiotic resistance profiles --- p.165 / "Appendix V Details of MRSA iolates selected for AFLP, Phage typing and MLST" --- p.166

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 4 uppsatser.

A study to assess the potentials of some commensal bacteria that belong to Staphylococcus, Acinetobacter and Stenotrophomonas genera as reservoirs of antibiotic resistance determinants in the environment of Nkonkobe Municipality of the Eastern Cape Province, South Africa, was carried out using standard microbiological and molecular techniques. A total of 120 Staphylococcus isolates which consisted of Staphylococcus haemolyticus (30%), Staphylococcus aureus (23.3%) from pig; Staphylococcus capitis (15%) from goat; Staphylococcus heamolyticus (5%) and Staphylococcus xylosus (15%) from cattle and other Staphylococci (11%) from dead chicken and pigs were isolated. About 23.3% of these isolates were coagulase positive and 76.7% were coagulase negative. This difference in prevalence along coagulase production divide was statistically significant (p < 0.05). Eighty-six Acinetobacter species (Acinetobacter baumannii/calcoaceticus and Acinetobacter haemolyticus) were also isolated from Alice and Fort Beaufort towns samples, while 125 Stenotrophomonas maltophilia isolates were from grass root rhizosphere (96%) and soil butternut root rhizosphere (4%). Between 75-100% of the Staphylococccus species were resistant to Penicillin G, tetracycline, sulphamethaxole and nalidixic acid; about 38 % were methicillin resistant, consisting of 12.6% methicillin resistant Staphylococcus aureus (MRSA) from pig and a total of 12% vancomycin resistant were observed. Also, 12% of the isolates were erythromycin resistant while 40.2 % were resistant to the third generation cephalosporin, ceftazidime. The antibiotic resistance genes vanA, VanB, eryA, eryB, eryC were not detected in all the phenotypically resistant Staphylococccus species, but mec A gene and mph genes were detected. In the Acinetobacter species, a wide range of 30-100% resistance to penicillin G, ceftriazone, nitrofurantoin, erythromycin, and augmentin was observed. Polymerase chain reaction (PCR) revealed the presence of Tet(B) and Tet(39) genes in these species, while Tet (A), Tet(M) and Tet(H) were absent. Also, 9.3% of the Acinetobacter species showed phenotypic production of extended spectrum beta lactamases (ESBLs) while 3.5% were positive for the presence of blaCTX-M-1 genes. The Stenotrophomonas maltophilia isolates showed varying resistance to meropenem (8.9%), cefuroxime (95.6 %), ampicillin-sulbactam (53.9%), ceftazidime (10.7%), cefepime (29.3 %), minocycline (2.2%), kanamycin (56.9%), ofloxacin (2.9%), levofloxacin (1.3%), moxifloxacin (2.8%), ciprofloxacin (24.3%), gatifloxacin (1.3%), polymyxin B (2.9 %), cotrimoxazole (26.1%), trimethoprim (98.6%), aztreonam(58%) and Polymyxin B (2.9 %). The isolates exhibited significant susceptibility to the fluoroquinolones (74.3-94.7 %), polymycin (97.1%) and meropenem (88.1%). Only sul3 genes were the only sulphonamide resistance gene detected among the trimethoprim-sulphamethoxazole resistant isolates. The observed multiple antibiotic resistance indeces (MARI) of >2 for Staphylococcus species, Acinetobacter species and Stenotrophomonas maltophilia suggest that they have arisen from high-risk sources where antibiotics are in constant arbitrary use resulting in high selective pressure. The presence of tetracycline resistance genes in Acinetobacter species justifies the observed phenotypic resistance to oxytetracycline and intermediate resistance to minocycline. High phenotypic resistance and the presence of some resistance genes in Staphylococcus species is a possible threat to public health and suggests animals to be important reservoirs of antibiotic resistance determinants in the environment. Indiscriminate use of antibiotics induces this kind of antibiotic resistance and should be discouraged. Personal hygiene is encouraged as it reduces the load of Acinetobacter species contacted from the environment that may be difficult to control. Commensal Stenotrophomonas maltophilia are as important as their clinical counterparts due to their roles in opportunistic infection, antibiotic resistance and their associated genes, especially sul gene. Personal hygiene is hereby advocated especially when in contact with soil, plants and plants’ rhizospheric soil

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Thesis (D.P.H.)--University of Hawaii at Manoa, 1991. / Includes bibliographical references (leaves 157-168). / Microfiche. / xiii, 168 leaves, bound ill. 29 cm

Thesis (M.Sc.) -- Memorial University of Newfoundland. / Typescript. Bibliography: leaves 80-86. Also available online.

Thesis (doctoral)--Utrecht, 1979.

It is the aim of this work to study the role, if any, of C'4 in the immune response against Staphylococcus aureus infections.

No description available.