Arthropod-Borne Infections in the United Kingdom and Saudi Arabia

Arthropod-Borne Infections in the United Kingdom and Saudi Arabia

Arthropod-borne infections in the United Kingdom and Saudi Arabia Bassam Alharbi Supervised by: Dr. Kevin Bown School of Environment and Life Sciences University of Salford, Salford, UK Submitted In Partial Fulfilment of the Requirement of the Degree of Doctor of Philosophy, March 2018. TABLE OF CONTENTS Table of tables. .................................................................................................................... VI Table of Figures. ................................................................................................................ VII Declaration............................................................................................................................ X Acknowledgment. ................................................................................................................ XI Abstract. ............................................................................................................................. XII Chapter One ........................................................................................................................... 1 Introduction and the aims of this thesis ................................................................................. 1 1. Introduction. ...................................................................................................................... 2 1.2. Protozoan parasites. .................................................................................................... 3 1.2.1 .Trypanosomiasis................................................................................................... 3 1.2.1.1. Host range of Trypanosoma. ......................................................................... 3 1.2.1.2. Importance of Trypanosomes. ....................................................................... 4 1.2.1.3. Transmission of human African trypanosomiasis and Chagas disease. ........ 6 1.2.1.4. The life cycle of Trypanosoma cruzi. ............................................................ 7 1.2.1.5. Animal Trypanosomiasis. .............................................................................. 8 1.2.1.6. Important trypanosomes in the United Kingdom. ......................................... 8 1.2.2.Theileria. ............................................................................................................. 12 1.2.2.1. Theileria life cycle. ...................................................................................... 14 1.2.2.2. Clinical manifestations of Theileriosis. ....................................................... 15 1.2.3. Babesia. .............................................................................................................. 16 1.2.3.1. Human Babesiosis. ...................................................................................... 17 1.2.3.2. Babesia life cycle......................................................................................... 18 1.2.3.3. Babesiosis in animals. ................................................................................. 19 1.2.3.4. Symptoms of Babesia infection. .................................................................. 21 1.2.4. Bartonella. .......................................................................................................... 21 1.2.4.1. Animal Bartonellosis. .................................................................................. 23 1.2.4.2. Host specificity. ........................................................................................... 27 1.2.5. Candidatus midichloria ...................................................................................... 28 1.3. Vector transmission. ................................................................................................. 29 1.3.1. Ticks. .................................................................................................................. 29 1.3.1.1. Hard Ticks. .................................................................................................. 29 1.3.1.2. The importance of the tick population on vertebrate health. ....................... 34 II 1.4. Investigated host. ...................................................................................................... 35 1.4.1. Bank vole (Myodes glareolus). .......................................................................... 35 1.4.2. Wood mice (Apodemus sylvaticus). ................................................................... 36 1.4.3. Libyan jird (Meriones libycus) ........................................................................... 37 1.4.4. Desert hedgehogs (Paraechinus aethiopicus). ................................................... 38 1.4.5. Red foxes (Vulpes vulpes). ................................................................................. 39 1.4.6. Greater white toothed shrew (Crocidura russula). ............................................ 40 1.4.7. Pygmy shrew (Sorex minutus) ................................................................................... 41 1.5. Aims and Objectives ................................................................................................. 42 Chapter Two ........................................................................................................................ 43 Materials and methods ......................................................................................................... 43 General Methods. ................................................................................................................ 44 2.1. Sample collection. ..................................................................................................... 44 2.2. DNA extraction. ........................................................................................................ 45 2.2.1. DNA extraction from rodent and red fox blood samples using a genomic DNA kit. ................................................................................................................................. 45 2.2.3. DNA extraction from FTA cards for Saudi jirds and hedgehogs. ...................... 45 2.3. Polymerase chain reaction. ....................................................................................... 46 2.3.1. Mammalian Tubulin PCR. ................................................................................. 46 2.3.2. Polymerase chain reaction (PCR). ..................................................................... 47 2.3.3. Real time PCR. ................................................................................................... 51 2.4. Gel electrophoresis. .................................................................................................. 53 2.5. Purification of PCR products. ................................................................................... 53 2.6. DNA concentration. .................................................................................................. 54 2.7. Bioinformatics software. ........................................................................................... 54 2.7.1. Finish TV............................................................................................................ 54 2.7.2. Blast Tool. .......................................................................................................... 55 2.7.3. Clustal omega. .................................................................................................... 55 2.7.4. Statistical analysis. ............................................................................................. 55 2.7.5. Phylogenetic tree. ............................................................................................... 55 2.8. Next generation sequencing for tick samples. .......................................................... 56 2.8.1. DNA extraction. ................................................................................................. 56 2.8.2. 16S rRNA Amplicon PCR conditions. ............................................................... 56 2.8.3. First PCR Clean-up. ........................................................................................... 57 III 2.8.4. Labelling of purified PCR products. .................................................................. 58 2.8.5. Purification of labelled 16SrRNA libraries (second clean up). .......................... 59 2.8.6. Quantification of 16S rRNA libraries. ............................................................... 59 2.8.7. Library Denaturing and Mi-seq sample loading. ............................................... 60 2.8.7.1. Denaturation and dilution of DNA. ............................................................. 60 2.8.7.2. Denaturation of diluted library. ................................................................... 61 2.8.7.3 .Denaturation and dilution of PhiX control and Amplicon Library combination. ............................................................................................................. 61 2.9. Bioinformatics analysis. ............................................................................................ 61 Chapter Three ...................................................................................................................... 62 Comparison of vector-borne protozoan infections in rodents from England and Ireland: the effect of invasion. ...............................................................................................................

View Full Text

Details

  • File Type
    pdf
  • Upload Time
    -
  • Content Languages
    English
  • Upload User
    Anonymous/Not logged-in
  • File Pages
    197 Page
  • File Size
    -

Download

Channel Download Status
Express Download Enable

Copyright

We respect the copyrights and intellectual property rights of all users. All uploaded documents are either original works of the uploader or authorized works of the rightful owners.

  • Not to be reproduced or distributed without explicit permission.
  • Not used for commercial purposes outside of approved use cases.
  • Not used to infringe on the rights of the original creators.
  • If you believe any content infringes your copyright, please contact us immediately.

Support

For help with questions, suggestions, or problems, please contact us