Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 6 uppsatser.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1980. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Chau Sze-lok. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 221-245). / Abstracts in English and Chinese. / ABSTRACT (English) --- p.i / ABSTRACT (Chinese) --- p.iii / ACKNOWLEDGMENT --- p.v / LIST OF CONTENTS --- p.vii / LIST OF TABLES --- p.xiv / LIST OF FIGURES --- p.xvii / ABBREVIATIONS --- p.xx / TERMS --- p.xxi / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / (PART A) / Chapter 1.1 --- Acinetobacter spp --- p.1 / Chapter 1.2 --- Clinical importance of Acinetobacter --- p.4 / Chapter 1.3 --- Resistance mechanisms / Chapter 1.3.1 --- Intrinsic resistance --- p.7 / Chapter 1.3.2 --- Acquired resistance --- p.15 / Chapter 1.4 --- Resistance in Acinetobacter --- p.21 / Chapter 1.4.1 --- The efflux system in Acinetobacter --- p.22 / Chapter 1.4.2 --- Other antibiotic resistance mechanisms in Acinetobacter --- p.23 / (PART B) / Chapter 1.5 --- Methods used in this study --- p.29 / Chapter 1.6 --- Rationale of this study --- p.35 / Chapter 1.7 --- Objectives --- p.37 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.39 / Chapter 2.1 --- Bacterial strains and isolates / Chapter 2.1.1 --- Isolates for studying blaIMP-4 --- p.39 / Chapter 2.1.2 --- Isolates for studying adeB --- p.39 / Chapter 2.1.3 --- Isolates for investigation of other efflux pump(s)in Acinetobacter GDG3 --- p.40 / Chapter 2.1.4 --- Isolates for studying the distribution of efflux pumps --- p.40 / Chapter 2.1.5 --- Reference strains --- p.41 / Chapter 2.2 --- Materials / Chapter 2.2.1 --- Sources of materials --- p.42 / Chapter 2.2.2 --- Buffers and solutions --- p.45 / Chapter 2.3 --- Instruments and software --- p.46 / Chapter 2.4 --- General bacteriological techniques / Chapter 2.4.1 --- Bacteriological dientification --- p.47 / Chapter 2.4.2 --- Stock isolates --- p.48 / Chapter 2.4.3 --- Retrieval of isolates --- p.48 / Chapter 2.5 --- General molecular biology techniques / Chapter 2.5.1 --- Agarose gel electrophoresis --- p.49 / Chapter 2.5.2 --- Polymerase chain reaction (PCR) --- p.50 / Chapter 2.5.3 --- Amplified Ribosomal Restriction DNA Analysis (ARDRA) --- p.51 / Chapter 2.5.4 --- Pulsed field gel electrophoresis (PFGE) --- p.53 / Chapter 2.5.5 --- Minimal inhibitory concentration (MIC) --- p.55 / Chapter 2.5.6 --- Antibiotic sensitivity test - disc diffusion test --- p.56 / Chapter 2.5.7 --- Detection of the presence of the common resistance mechanisms --- p.57 / Chapter 2.5.8 --- TA Cloning --- p.60 / Chapter 2.5.9 --- DNA Sequencing --- p.62 / Chapter 2.5.10 --- Sequence analysis --- p.64 / Chapter 2.5.11 --- CYBR Green Assay --- p.65 / Chapter 2.5.12 --- Complementary DNA (cDNA) preparation --- p.66 / Chapter 2.5.13 --- Real time RT-PCR --- p.67 / Chapter 2.5.14 --- Construction of Genome Walker Libraries --- p.69 / Chapter 2.6 --- "Selection of acinetobacters from ICU, blood culture and other clinical isolates" / Chapter 2.6.1 --- Isolates from existing stock cultures --- p.71 / Chapter 2.6.2 --- New isolates obtained for this study --- p.71 / Chapter 2.7 --- Study of expression level of the blaIMP-4 gene / Chapter 2.7.1a --- Verification of the specificity of primers for blaIMP-4 --- p.72 / Chapter 2.7.1b --- Verfication of the specificity of primers for 16S rRNA gene --- p.73 / Chapter 2.7.1c --- Construction of standard curve --- p.76 / Chapter 2.7.2 --- Expression levels of blaIMP-4 and meropenem MICin blaIMP-4+ blood culture isolates --- p.77 / Chapter 2.7.3 --- Intra-assay reproducibility --- p.11 / Chapter 2.7.4 --- Detection of the production of metallo-β-lactamase --- p.77 / Chapter 2.8 --- Study of adeABC expression / Chapter 2.8.1 --- Determination of the presence of the adeB gene --- p.78 / Chapter 2.8.2 --- Entirety of the adeABC operon --- p.79 / Chapter 2.8.3 --- Expression level of the adeB gene --- p.82 / Chapter 2.8.4 --- Expression levels of adeB in sets of serial isolates --- p.84 / Chapter 2.8.5 --- Intra-assay reproducibility --- p.84 / Chapter 2.8.6 --- Inter-assay reproducibility --- p.84 / Chapter 2.9 --- Investigation of other efflux pumps in acinetobacter genomic DNA group3 / Chapter 2.9.1 --- Detection of adeB homologue in a genomic DNA group 3isolate --- p.85 / Chapter 2.9.2 --- Chromosome walking of the adeB-like genes --- p.87 / Chapter 2.9.3 --- Sequences of AdeE and AdeY and their comparison --- p.105 / Chapter 2.9.4 --- Topology prediction of AdeE and AdeY --- p.105 / Chapter 2.9.5 --- The role of the putative pump AdeE --- p.106 / Chapter 2.10 --- Distribution of AdeB and the putative efflux pumps AdeE and AdeY in acinetobacters from different bacterial collections / Chapter 2.10.1 --- Distribution of adeB and the putative pumps (adeE and adeY) in blood cultures (1997-2000) --- p.113 / Chapter 2.10.2 --- Confirmation of the identity of the amplification products of adeE and ade Y in blood culture isolate (1997-2000) --- p.115 / Chapter 2.10.3 --- The presence of adeE in GDG 3 acinetobacters from different sources --- p.116 / Chapter 2.10.4 --- "The presence of adeB, adeE and adeY in antibiotic susceptibility" --- p.116 / Chapter 2.10.5 --- "adeB, adeE and adeY and the clonally and epidemiologically related sets of isolates" --- p.116 / Chapter 2.10.6 --- "adeB, adeE and adeY and the blaIMP-4+ isolates" --- p.116 / Chapter CHAPTER 3 --- "SELECTION OF ACINETOBACTERS FROM ICU, BLOOD CULTURE AND OTHER CLINICAL ISOLATES" --- p.119 / Chapter 3.1 --- Results / Chapter 3.1.1 --- Isolates from existing stock cultures --- p.119 / Chapter 3.1.2 --- New isolates obtained for this study --- p.127 / Chapter 3.2 --- Discussion / Chapter 3.2.1 --- Identification of clonally related isolates by PFGE --- p.129 / Chapter 3.2.2 --- Correlation between the presence of common resistance mechanisms and the changes in antimicrobial susceptibility --- p.129 / Chapter 3.2.3 --- Development of resistance in serial isolates --- p.131 / Chapter CHAPTER 4 --- STUDY OF blaIMP-4 EXPRESSION --- p.133 / Chapter 4.1 --- Results / Chapter 4.1.1 --- Study of expression level of the blaIMP-4 gene --- p.134 / Chapter 4.1.2 --- Expression levels of blaIMP-4 and meropenem MIC in blaIMP-4+ blood culture isolates --- p.136 / Chapter 4.1.3 --- Intra-assay reproducibility \ --- p.37 / Chapter 4.1.4 --- Detection of the production of metallo-β-lactamase --- p.140 / Chapter 4.2 --- Discussion / Chapter 4.2.1 --- Dissociation curve --- p.140 / Chapter 4.2.2 --- Reproducibility of real time RT-PCR --- p.140 / Chapter 4.2.3 --- Relationship between mRNA level of blaIMP-4 and the meropenem MIC --- p.142 / Chapter CHAPTER 5 --- STUDY OF adeABC EXPRESSION --- p.145 / Chapter 5.1 --- Results / Chapter 5.1.1 --- Determination of the presence of the adeB gene --- p.145 / Chapter 5.1.2 --- Entirety of the adeABC operon --- p.146 / Chapter 5.1.3 --- Expression level of the adeB gene --- p.148 / Chapter 5.1.4 --- Expression levels of adeB in sets of serial isolates --- p.151 / Chapter 5.1.5 --- Intra-assay reproducibility --- p.154 / Chapter 5.1.6 --- Inter-assay reproducibility --- p.154 / Chapter 5.2 --- Discussion / Chapter 5.2.1 --- Detection of adeB --- p.156 / Chapter 5.2.2 --- Entirety of the adeABC operon --- p.156 / Chapter 5.2.3 --- Reproducibility of real time RT-PCR --- p.157 / Chapter 5.2.4 --- Relationship between adeB-mRNA level and antimicrobial susceptibility --- p.157 / Chapter CHAPTER 6 --- INVESTIGATION OF OTHER EFFLUX PUMPS IN ACINETOBACTER GENOMIC DNA GROUP3 --- p.159 / Chapter 6.1 --- Results / Chapter 6.1.1 --- Detection of adeB homologue in a genomic DNA group3 isolate --- p.159 / Chapter 6.1.2 --- Chromosome walking of the adeB-like genes --- p.162 / Chapter 6.1.3 --- Sequences of AdeE and AdeY and their comparison --- p.173 / Chapter 6.1.4 --- Topology prediction of AdeE and AdeY --- p.175 / Chapter 6.1.5 --- The role of the putative pump AdeE --- p.177 / Chapter 6.2 --- Discussion / Chapter 6.2.1 --- The AdeE RND transporter --- p.181 / Chapter 6.2.2 --- The theoretical AdeY protein --- p.183 / Chapter CHAPTER 7 --- DISTRIBUTION OF AdeB AND THE PUTATIVE EFFLUX PUMPS AdeE and AdeY IN ACINETOBACTERS FROM DIFFERENT BACTERIAL COLLECTIONS --- p.184 / Chapter 7.1 --- Results / Chapter 7.1.1 --- Distribution of adeB and the putative pumps (adeE and ade Y) in blood cultures (1997-2000) --- p.184 / Chapter 7.1.2 --- Confirmation of the identity of the amplification products of adeE and adeY in blood culture isolates (1997-2000) --- p.187 / Chapter 7.1.3 --- The presence of adeE in GDG 3 acinetobacters from different sources --- p.195 / Chapter 7.1.4 --- "The presence of adeB, adeE and ade Y in antibiotic susceptibility" --- p.196 / Chapter 7.1.5 --- "adeB, adeE and adeY and the clonally and epidemiologically related sets of isolates" --- p.202 / Chapter 7.1.6 --- "adeB, adeE and adeY and the blaIMP-4+ isolates" --- p.202 / Chapter 7.2 --- Discussion / Chapter 7.2.1 --- PCR-RFLP typing --- p.205 / Chapter 7.2.2 --- Distribution of adeB --- p.205 / Chapter 7.2.3 --- Distribution of adeE --- p.206 / Chapter 7.2.4 --- Distribution of adeY --- p.207 / Chapter 7.2.5 --- Distribution of adeE and adeY in GDG 3 isolates --- p.207 / Chapter CHAPTER 8 --- GENERAL DISCUSSION --- p.209 / Chapter 8.1 --- Significance of adeB and the putative pumps (adeE and adeY) --- p.211 / Chapter CHAPTER 9 --- CONCLUSION --- p.218 / Chapter 9.1 --- Conclusion --- p.218 / Chapter 9.2 --- Future Plan --- p.219 / REFERENCES --- p.221 / APPENDIX --- p.246 / Appendix1 --- p.246 / Appendix2 --- p.247 / Appendix3 --- p.252 / Appendix4 --- p.253 / Appendix5 --- p.259

The Enterobacter are often found in the feces of man and other warm blooded animals but have limited use as indicators of fecal contamination of foods and water since they are also often found as natural flora of non-fecal environments such as water and plants. This study proposes a method that may be useful for differentiating between fecal and non-fecal Enterobacter. Coliform bacteria associated with the human fecal environment often have a high incidence of multiple resistance to antibiotics. By determining multiple drug resistance and indexing Enterobacter in reference to their environmental source, it was discovered that isolates from human feces, raw sewage and certain foods demonstrated a higher index than isolates from rural soils, wild animal feces, cereal grains and other non-human environments. An index value greater than 0.2 indicated isolates from environments contaminated with human feces representing a high health risk while those isolates with an index of less than 0.2 came from relatively safe environments. Traditionally, the assessment of food borne health hazards is determined by measuring the quantity and kind of bacteria present. Drug resistance indexing will compliment these tests and aid in identifying serious bacterial contamination of foods. / Graduation date: 1981

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Antimicrobials have been instrumental in the treatment of infectious disease: responsible for worldwide infection control and reductions in disease-induced morbidity, and mortality. However, in every case where new chemotherapeutic agents have been introduced, resistance to them has eventually evolved. Principally, the current strategy for dealing with this problem is to invest heavily in drug development, with the hope that new drugs become available before all existing drugs lose their efficacy. Instead of focusing on the ‘development side’ of the problem, another possible strategy is to invest in methods of slowing evolution of resistance. We use a novel application of queueing theory to demonstrate that, when comparing equivalent changes in drug development versus evolution management, the latter has a much greater effect on ensuring a continued supply of effective antimicrobial agents. Our results therefore call for a reappraisal of the current emphasis on enhancing drug development as a means of managing resistance. / Thesis (Master, Biology) -- Queen's University, 2013-07-02 15:23:55.906

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