BIODIVERSITY OF THE MUSCIDAE (DIPTERA) FROM CHURCHILL, MANITOBA, CANADA, WITH TAXONOMIC ISSUES REVEALED OR RESOLVED BY DNA BARCODING by Anaïs Krystel Renaud A thesis submitted to the Faculty of Graduate Studies of The University of Manitoba in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Entomology University of Manitoba Winnipeg, MB Copyright © December 2011 by Anaïs Krystel Renaud TABLE OF CONTENTS ACKNOWLEDGMENTS ................................................................................................... i DEDICATION ................................................................................................................... iii ABSTRACT ....................................................................................................................... iv LIST OF FIGURES ............................................................................................................ v LIST OF TABLES ........................................................................................................... viii LIST OF APPENDICES ..................................................................................................... x CHAPTER 1 - GENERAL INTRODUCTION .............................................................. 1 CHAPTER 2 - LITERATURE REVIEW ...................................................................... 3 THE ARCTIC AND SUBARCTIC ECOREGIONS ...................................................... 3 Hudson Bay Lowlands ................................................................................................ 4 Churchill ..................................................................................................................... 4 ENVIRONMENTAL CHANGES IN THE NORTH: A FOCUS ON CHURCHILL (MB) ................................................................................................................................ 5 Environmental impacts of human activities in Churchill ........................................... 6 INSECTS OF CHURCHILL IN A CONTEXT OF ENVIRONMENTAL CHANGE .. 8 Responses to climate change ...................................................................................... 9 Responses to habitat modifications ........................................................................... 10 MUSCIDAE .................................................................................................................. 11 Ecology and natural history ...................................................................................... 13 Medical and economic importance ........................................................................... 16 Dispersal ................................................................................................................... 16 Additional information.............................................................................................. 17 INVENTORIES OF NORTHERN INSECTS .............................................................. 17 Historical records of the Muscidae of Churchill ....................................................... 18 Current diversity of the Muscidae of Churchill ........................................................ 19 SAMPLING MUSCIDAE ............................................................................................ 19 The sweep net ........................................................................................................... 20 The pan trap .............................................................................................................. 20 The Malaise trap ....................................................................................................... 21 SPECIMEN PREPARATION ...................................................................................... 22 SPECIMEN IDENTIFICATION .................................................................................. 23 MEASURING DIVERSITY ......................................................................................... 27 APPLICATIONS AND LIMITATIONS OF MUSEUM AND LITERATURE DATA FOR COMPARATIVE PURPOSES ............................................................................ 30 CHAPTER 3 - MUSCIDAE (DIPTERA) DIVERSITY IN CHURCHILL, MANITOBA, CANADA, BETWEEN TWO TIME PERIODS: EVIDENCE FOR LIMITED CHANGES SINCE 1965 .............................................................................. 33 INTRODUCTION ........................................................................................................ 35 MATERIALS & METHODS ....................................................................................... 39 2007 data set ............................................................................................................. 39 Pre-1965 data set ....................................................................................................... 41 Taxonomic identification .......................................................................................... 42 Data analyses ............................................................................................................ 43 RESULTS ..................................................................................................................... 47 2007 survey ............................................................................................................... 47 Pre-1965 data set ....................................................................................................... 48 Historical vs contemporary data sets comparisons ................................................... 49 DISCUSSION ............................................................................................................... 52 CHAPTER 4 - DNA BARCODING OF THE MUSCIDAE (DIPTERA) OF CHURCHILL, MANITOBA, CANADA, REVEALS HIGH CORRESPONDENCE BETWEEN MORPHOLOGICAL AND MOLECULAR SPECIES LIMITS .......... 71 ABSTRACT .................................................................................................................. 72 INTRODUCTION ........................................................................................................ 73 What is DNA barcoding............................................................................................ 73 Species identification with DNA barcoding ............................................................. 75 Muscidae ................................................................................................................... 77 Objectives ................................................................................................................. 80 MATERIAL & METHODS ......................................................................................... 81 Specimen selection.................................................................................................... 81 DNA processing and alignment ................................................................................ 81 Data analysis ............................................................................................................. 84 RESULTS ..................................................................................................................... 87 Taxonomic reassessment .......................................................................................... 90 DISCUSSION ............................................................................................................... 93 DNA amplification and sequencing process ............................................................. 93 DNA barcoding as an efficient tool for identification of muscid flies ..................... 94 Intraspecific distance ................................................................................................ 96 Interspecific distance ................................................................................................ 99 Conclusion .............................................................................................................. 100 CHAPTER 5 - GENERAL DISCUSSION ................................................................. 109 RECOMMENDATIONS FOR FUTURE INVENTORIES ....................................... 109 LIMITATIONS IN THE COMPARISON OF HISTORICAL VS CONTEMPORARY ASSEMBLAGES ........................................................................................................ 112 OVERLAP IN SPECIES RICHNESS BETWEEN TIME PERIODS: THE RESILIENCE OF MUSCIDAE .................................................................................. 113 NON-OVERLAPPING SPECIES .............................................................................. 115 DNA BARCODING ................................................................................................... 116 CONSERVATION ..................................................................................................... 118 LITERATURE CITED ................................................................................................... 119 APPENDICES ................................................................................................................ 156 ACKNOWLEDGMENTS The completion of this thesis was only possible because of the invaluable support and constant guidance of my co-supervisor Dr. Jade Savage who devoted her time and energy to ensure the achievement of the project. I would also like to acknowledge the support of my two other co-supervisors, Dr. Rob Roughley and Dr. Terry Galloway (interim),