UCC Library and UCC researchers have made this item openly available. Please let us know how this has helped you. Thanks! Title Epidemiology and comparative analysis of Yersinia in Ireland Author(s) Ringwood, Tamara Publication date 2013 Original citation Ringwood, T. 2013. Epidemiology and comparative analysis of Yersinia in Ireland. PhD Thesis, University College Cork. Type of publication Doctoral thesis Rights © 2013, Tamara Ringwood http://creativecommons.org/licenses/by-nc-nd/3.0/ Item downloaded http://hdl.handle.net/10468/1294 from Downloaded on 2021-10-07T12:07:10Z Epidemiology and Comparative Analysis of Yersinia in Ireland by Tamara Ringwood A thesis presented for the Degree of Doctor of Philosophy National University of Ireland, Cork University College Cork Coláiste na hOllscoile Corcaigh Department of Microbiology Head of Department: Prof. Gerald F. Fitzgerald Supervisor: Prof. Michael B. Prentice April 2013 Contents List of Tables .............................................................................................................................................. iv List of figures ............................................................................................................................................. vi Declaration ............................................................................................................................................... viii Acknowledgements ................................................................................................................................. ix Abbreviations .............................................................................................................................................. x Abstract ...................................................................................................................................................... xiii CHAPTER 1 LITERATURE REVIEW 15 1.1 Epidemiology ............................................................................................................................ 16 1.1.1 Incidence rates of enteric yersiniosis in humans ................................................................. 16 1.1.2 Risk factors and transmission of enteropathogenic Yersinia .......................................... 18 1.2 Isolation and characterization of Y. enterocolitica and Y. pseudotuberculosis . 21 1.2.1 Yersinia selective enrichment procedures .............................................................................. 21 1.2.2 Alkali treatment .................................................................................................................................. 22 1.2.3 Yersinia selective agar medium .................................................................................................... 22 1.2.4 Biochemical identification of enteropathogenic Yersinia species ................................. 24 1.2.5 Biotyping of Y. enterocolitica ......................................................................................................... 26 1.2.6 Biotyping of Y. pseudotuberculosis .............................................................................................. 27 1.2.7 Serotyping of Y. enterocolitica and Y. pseudotuberculosis................................................. 27 1.3 Molecular detection and characterization of Yersinia ............................................... 27 1.3.1 Molecular detection of pathogenic Yersinia species ........................................................... 27 1.3.2 Molecular identification of Yersinia species ........................................................................... 29 1.4 Clinical manifestations of yersiniosis .............................................................................. 32 1.4.1 Gastrointestinal manifestations of yersiniosis ...................................................................... 32 1.4.2 Extraintestinal manifestations of yersiniosis ........................................................................ 33 1.4.3 Yersinia and blood transfusion ..................................................................................................... 34 1.4.4 Post infectious complications ....................................................................................................... 34 1.5 Immunology of yersiniosis ................................................................................................... 35 1.5.1 Immune response during Yersinia infection .......................................................................... 35 1.5.2 Anti-Yersinia seroprevalence ........................................................................................................ 35 1.6 Virulence factors of enteropathogenic Yersinia species ........................................... 38 1.6.1 Chromosome encoded virulence factors ................................................................................. 38 1.6.2 Plasmid encoded virulence factors ............................................................................................. 44 1.6.3 Novel virulence factors .................................................................................................................... 48 1.7 Genotyping of Yersinia species ........................................................................................... 49 1.7.1 Multilocus Enzyme Electrophoresis (MLEE) ......................................................................... 49 1.7.2 Multi Locus Sequence Typing (MLST) ....................................................................................... 50 1.7.3 Repetitive elements PCR ................................................................................................................. 51 1.7.4 Ribotyping ............................................................................................................................................. 51 1.7.5 Pulse Field Gel Electrophoresis (PFGE) .................................................................................... 52 i 1.7.6 Amplified Fragment Length Polymorphism (AFLP) ........................................................... 53 1.7.7 Multiple Locus Variable number tandem repeat Analysis (MLVA) ............................. 54 1.7.8 Microarray Genotyping .................................................................................................................... 54 1.8 Evolution of Yersinia species ............................................................................................... 56 1.9 Aims and Objectives ............................................................................................................... 59 CHAPTER 2 CURRENT EVIDENCE FOR HUMAN YERSINIOSIS IN IRELAND 61 2.1 Introduction .............................................................................................................................. 62 2.2 Materials and methods .......................................................................................................... 63 2.2.1 Clinical specimens screening methodology ............................................................................ 63 2.2.2 Recovery of Y. enterocolitica from experimentally inoculated faecal specimens .. 65 2.2.3 Yersinia identification procedures .............................................................................................. 66 2.2.4 Human sewage and pig slurry screening for pathogenic Yersinia ................................ 66 2.2.5 Serological screening ........................................................................................................................ 69 2.2.6 Statistical analysis .............................................................................................................................. 70 2.3 Results ......................................................................................................................................... 70 2.3.1 Clinical specimens screening ........................................................................................................ 70 2.3.2 Human sewage and pig slurry screening for pathogenic Yersinia ................................ 73 2.3.3 Serology .................................................................................................................................................. 78 2.4 Discussion .................................................................................................................................. 79 CHAPTER 3 YERSINIA INSECT TOXICITY 83 3.1 Introduction .............................................................................................................................. 84 3.1.1 Genetic organisation of Yersinia insecticidal toxin complexes ...................................... 84 3.1.2 Insect models ....................................................................................................................................... 86 3.1.3 Biological activity of insecticidal toxins ................................................................................... 86 3.1.4 Cytolethal distending toxin ............................................................................................................ 89 3.1.5 Toxicity of Y. enterocolitica strains isolated in Ireland for G. mellonella ................... 90 3.2 Materials and Methods .......................................................................................................... 90 3.2.1 Yersinia strains ...................................................................................................................................