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Eastern Bat Bug (Cimex Adjunctus), and Swallow Bug (Oeciacus Vicarius)

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Comparison of the Mitochondrial Genomes of the Common Bed Bug (Cimex lectularius), Eastern Bat Bug (Cimex adjunctus), and Swallow Bug (Oeciacus vicarius) A thesis submitted to the College of Graduate Studies and Research in Partial Fulfillment of the Requirements for the degree of Master of Science in the Department of Biology University of Saskatchewan Saskatoon, Saskatchewan, Canada By James S. Armstrong © Copyright James S. Armstrong, July 2015. All rights reserved PERMISSION TO USE In presenting this thesis/dissertation in partial fulfillment of the requirements for a Postgraduate degree from the University of Saskatchewan, I agree that the Libraries of this University may make it freely available for inspection. I further agree that permission for copying of this thesis/dissertation in any manner, in whole or in part, for scholarly purposes may be granted by the professor or professors who supervised my thesis/dissertation work or, in their absence, by the Head of the Department or the Dean of the College in which my thesis work was done. It is understood that any copying or publication or use of this thesis/dissertation or parts thereof for financial gain shall not be allowed without my written permission. It is also understood that due recognition shall be given to me and to the University of Saskatchewan in any scholarly use which may be made of any material in my thesis/dissertation. i DISCLAIMER Reference in this thesis to any specific commercial products, process, or service by trade name, trademark, manufacturer, or otherwise, does not constitute or imply its endorsement, recommendation, or favoring by the University of Saskatchewan. The views and opinions of the author expressed herein do not state or reflect those of the University of Saskatchewan, and shall not be used for advertising or product endorsement purposes. Requests for permission to copy or to make other uses of materials in this thesis/dissertation in whole or part should be addressed to: Head of the Department of Biology University of Saskatchewan 112 Science Place Saskatoon, Saskatchewan S7N 5E2 Canada ii ABSTRACT Species within the family Cimicidae (bed bugs) are hematophagous ectoparasites of mammals and birds. Many cimicids are of socio-economic importance. Despite the global resurgence of these pests, there is currently a paucity of information regarding the mitochondrial (mt) DNA sequences of cimicids. Therefore, I used a PCR-based primer walking strategy to amplify and sequence the near complete mitogenome of the common bed bug (Cimex lectularius), and several mitochondrial gene regions of the Eastern bat bug (Cimex adjunctus) and swallow bug (Oeciacus vicarius). I compared the mitochondrial genetic variability between C. lectularius from two populations to look for molecular markers useful for population genetic studies. Furthermore, the mt DNA sequences of these species of medical and veterinary importance were compared to those of other heteropterans to infer the evolutionary relationships of species in the family Cimicidae. iii ACKNOWLEDGEMENTS I would like to thank my supervisor, Dr. Neil Chilton for his guidance and support through both my B.Sc and M.Sc research. His expertise and mentorship during the past four years has been greatly appreciated. I would also like to thank Dr. Art Davis, Dr. Jose Andres, Dr. Dwayne Hegedus for acting as my advisory committee and external examiner. Within the department I would like to acknowledge Dr. Carlos Carvalho and Kunal Baxi for assisting me with photographs. Lastly, I would like to thank my lab mates, particularly Dr. Clare Anstead, who oversaw my training in lab techniques. iv TABLE OF CONTENTS PERMISSION TO USE ................................................................................................. i DISCLAIMER ................................................................................................................ ii ABSTRACT .................................................................................................................... iii ACKNOWLEDGEMENTS .......................................................................................... iv TABLE OF CONTENTS .............................................................................................. v LIST OF TABLES ......................................................................................................... viii LIST OF FIGURES ........................................................................................................ x LIST OF ABBREVIATIONS ....................................................................................... xii 1. INTRODUCTION .................................................................................................... 1 1.1 Biology of Cimex lectularius and other cimicids .............................................. 1 1.1.1 Overview ................................................................................................... 1 1.1.2 Finding and feeding from hosts ................................................................ 2 1.1.3 Reproduction and life cycle ...................................................................... 3 1.2 Significance of Cimex lectularius ...................................................................... 5 1.3 Global resurgence of Cimex lectularius ............................................................. 6 1.4 Population dynamics and genetics of Cimex lectularius ................................... 9 1.4.1 Population composition ............................................................................. 9 1.4.2 Dispersal .................................................................................................... 9 1.4.3 Population genetics.................................................................................... 10 1.5 Cimex adjunctus and Oeciacus vicarius ............................................................ 11 v 1.6 Systematics of the family Cimicidae ................................................................. 12 1.7 Mitochondrial DNA and mitogenomics ............................................................ 13 1.8 Objectives of this thesis ...................................................................................... 14 2. MATERIALS AND METHODS ............................................................................. 1 7 2.1 Cimex lectularius ................................................................................................ 17 2.1.1 Collection and identification of specimens .............................................. 17 2.1.2 DNA extraction ......................................................................................... 20 2.1.3 Molecular identification of bed bugs ........................................................ 23 2.1.4 Amplification and characterization of the mt genome ............................. 23 2.1.5 DNA sequence analyses ............................................................................ 30 2.2 Cimex adjunctus.................................................................................................. 33 2.2.1 Collection and identification of specimens .............................................. 33 2.2.2 Amplification and characterization of the mt genome ............................. 35 2.3 Oeciacus vicarius................................................................................................ 37 2.3.1 Collection and identification of specimens .............................................. 37 2.3.2 Amplification and characterization of the mt genome ............................. 39 3. RESULTS ................................................................................................................. 41 3.1 Mitochondrial DNA sequence of Cimex lectularius ........................................ 41 3.1.1 Mitogenome composition and organization ............................................. 41 3.1.2 Protein coding genes ................................................................................ 46 3.1.3 Ribosomal RNA and transfer RNA genes ................................................ 47 3.1.4 Gene regions with intraspecific variation ................................................. 49 3.1.5 Phylogenetic relationships of Cimex lectularius ...................................... 50 3.2 Mitochondrial DNA sequence of Cimex adjunctus ........................................... 54 3.2.1 Mitogenome composition and organization ............................................. 54 3.2.2 Protein coding genes ................................................................................ 54 3.2.3 Ribosomal RNA and transfer RNA genes ................................................ 54 3.2.4 Phylogenetic relationships of Cimex adjunctus ........................................ 57 vi 3.3 Mitochondrial DNA sequence of Oeciacus vicarius ......................................... 59 3.3.1 Mitogenome composition and organization ............................................. 59 3.3.2 Protein coding genes ................................................................................ 59 3.3.3 Ribosomal RNA and transfer RNA genes ................................................ 59 3.3.4 Phylogenetic relationships of Oeciacus vicarius ...................................... 64 3.4 Phylogenetic relationships of Cimex lectularius, Cimex adjunctus Oeciacus vicarius ............................................................................................... 66 4. DISCUSSION .........................................................................................................
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