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Floral and honeydew foraging ecology of select mosquito species by Daniel Peach B.Sc. (Dist.), Simon Fraser University, 2012 Associate of Arts, Douglas College, 2008 Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the Department of Biological Sciences Faculty of Science © Daniel Peach 2019 SIMON FRASER UNIVERSITY Spring 2019 Copyright in this work rests with the author. Please ensure that any reproduction or re-use is done in accordance with the relevant national copyright legislation. Approval Name: Daniel Peach Degree: Doctor of Philosophy Title: Floral and honeydew foraging ecology of select mosquito species Examining Committee: Chair: Jim Mattsson Associate Professor Gerhard Gries Senior Supervisor Professor Carl Lowenberger Supervisor Professor Margo Moore Supervisor Professor Emerita Sheila Fitzpatrick Examiner Research Scientist Agriculture and Agri-Food Canada Woodbridge Foster External Examiner Faculty Emeritus Department of Entomology The Ohio State University Date Defended/Approved: April 26, 2019 ii Abstract Both male and female mosquitoes exploit a wide variety of plant sugar resources, including floral nectar and aphid honeydew, as important sources of carbohydrates. Mosquitoes are generally considered nectar thieves that do not pollinate the flowers they visit, and volatile semiochemicals are believed to be the primary driver of mosquito attraction to plant sugar sources. Using the northern house mosquito, Culex pipiens, and its nectar host the common tansy, Tanacetum vulgare, we showed mosquito-induced seed-set. We found that semiochemicals from T. vulgare flowers are attractive to Cx. pipiens and the yellow fever mosquito, Aedes aegypti, that visual and olfactory inflorescence cues in combination attract more mosquitoes than olfactory cues alone, and that plant CO2 enhances the attractiveness of a 20-component synthetic blend of tansy inflorescence odourants. This blend included 9 odourants found in human odour, which are also attractive. Electroretinograms revealed that Cx. pipiens eyes can sense ultra-violet (UV) wavelengths, with peak sensitivity at 335 nm. Experiments found that UV inflorescence cues of T. vulgare and the common hawkweed, Hieracium lachenalii, enhance the attractiveness of inflorescence odour to female Cx. pipiens through floral patterns of UV-absorption and UV-reflection. We then established the attraction of Ae. aegypti to honeydew odourants from the green peach aphid, Myzus persicae, and the pea aphid, Acyrthosiphon pisum, feeding on fava bean, Vicia faba. We collected and analyzed headspace odourants from honeydew of A. pisum feeding on V. faba. An 8- component synthetic blend of these odourants and synthetic odourant blends of crude and sterile honeydew we prepared from literature data all attracted female Ae. aegypti. The synthetic blend containing microbial odour constituents proved more effective than the blend without these constituents. Our data support the hypotheses that mosquitoes are pollinators, that the entire inflorescence Gestalt of olfactory, CO2 and UV cues is more attractive to mosquitoes than floral odourants alone, that olfactory cues attract mosquitoes to honeydew, and that microbe-emitted volatiles play a role in mosquito attraction to honeydew. Keywords: Mosquitoes; Aedes aegypti; Culex pipiens; Pollination; Chemical Ecology; Sensory Ecology iii Acknowledgements I give my deepest gratefulness and appreciation to my advisor, Dr. Gerhard Gries, for support, guidance, and a shared fantastic sense of humour. He is an amazing person and mentor who has immensely aided me in my development as a scientist, teacher, and communicator. Thank you for your encouragement, collegiality, being open to unorthodox ideas, the passion you enkindle in others, and the opportunities you have provided me with. I also express sincere gratitude to Dr. John Borden and to Regine Gries for so much lively and productive discussion, mentorship, and constructive criticism. Your guidance, whether solicited or unsolicited, is always welcome. I extend thanks to my committee members for their support and advice. Dr. Carl Lowenberger for his insightful (and often humbling) questions and critiques, which have kept me on my toes with a smile on my face. Dr. Margo Moore for her piercing questions, unwavering optimism, and encouragement and embracement of my zany ideas. I thank Dr. Jim Mattsson for chairing the committee. I also thank Dr. Sheila Fitzpatrick for agreeing to serve as my internal examiner, and Dr. Woodbridge Foster for agreeing to serve as my external examiner. My research was assisted by the contributions of several undergraduate researchers that worked on many of the projects associated with this dissertation. I thank them all for their hard work! Specifically, I thank Elton Ko (now a graduate student), Nathan Young, Sanam Meraj, Max Almond, Sujani Gomes, April Balcita, and Leon Li. I would also like to thank my mentors and friends Dr. Peter Belton, Elspeth Belton, Karen Needham, Tammy McMullan, and Dr. Sean McCann for their guidance over the years and for always listening to, and encouraging, my rants about mosquitoes and other topics. The members of the Gries lab have contributed immensely to my professional development. Thank you all for providing thought-provoking conversations, amazing collaborations, and for such a cheerful and supportive research environment! Regine Gries provided invaluable guidance and tutorship on the chemistry-related aspects of my research. Dr. Huiman Zhai and Dr. Santosh Kumar Alamsetti synthesized many of the iv chemicals needed for my research. I would also specifically like to thank current and former lab-mates Adam Blake, Elton Ko, Catherine Scott, Dr. Bekka Brodie, Dr. Sean McCann, Josh Pol, Asim Renyard, Yonathan Uriel, and Jaime Chalissery. I thank my friend, colleague, and co-author Adam Blake for statistical and visual ecology advice, for amazing photography, for sharing his excellent taste in beer, and for being such an all-around fantastic person. I thank my family for their patience and support throughout my entire graduate career. Thank you for always being there for me and embracing my wackiness. My most heartfelt thanks to my beautiful and talented fiancé (and informal research assistant), Grace, for her love, support, encouragement, and the sacrifices she has made on my behalf during the pursuit of my PhD. v Table of Contents Approval .......................................................................................................................... ii Abstract .......................................................................................................................... iii Acknowledgements ........................................................................................................ iv Table of Contents ........................................................................................................... vi List of Tables ................................................................................................................... x List of Figures................................................................................................................. xi List of Acronyms ........................................................................................................... xvi Chapter 1. Mosquito phytophagy – sources exploited, ecological function, and evolutionary transition to haematophagy1 ......................................................... 1 Introduction ..................................................................................................................... 1 1.1. Is the term “sugar-feeding” appropriate? ................................................................ 2 1.1.1. Floral nectar .................................................................................................. 2 1.2. Plant-derived food sources sought and consumed by mosquitoes ......................... 5 1.2.1. Floral nectar .................................................................................................. 5 1.2.2. Extrafloral/extrasoral nectaries ...................................................................... 7 1.2.3. Fruit and seedpods ........................................................................................ 8 1.2.4. Plant tissues .................................................................................................. 9 1.2.5. Honeydew ................................................................................................... 10 1.2.6. Ant regurgitate ............................................................................................. 11 1.2.7. Sweet food waste ........................................................................................ 11 1.2.8. Summary ..................................................................................................... 12 1.3. Do mosquitoes have a functional role as pollinators? .......................................... 12 1.3.1. Instances of nectar-theft .............................................................................. 12 1.3.2. Evidence for pollination ................................................................................ 13 1.3.3. Summary ..................................................................................................... 14 1.4. Predation risk of plant-foraging mosquitoes ......................................................... 14 1.5. The role of microbes in attracting mosquitoes