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Viral Prevalence Among Social Bees in Different Landscapes Tugce Karacoban University of Nebraska - Lincoln

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Viral Prevalence Among Social Bees in Different Landscapes Tugce Karacoban University of Nebraska - Lincoln University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Dissertations and Student Research in Entomology Entomology, Department of 12-2018 Viral prevalence among social bees in different landscapes Tugce Karacoban University of Nebraska - Lincoln Follow this and additional works at: http://digitalcommons.unl.edu/entomologydiss Part of the Entomology Commons, and the Immunology and Infectious Disease Commons Karacoban, Tugce, "Viral prevalence among social bees in different landscapes" (2018). Dissertations and Student Research in Entomology. 56. http://digitalcommons.unl.edu/entomologydiss/56 This Article is brought to you for free and open access by the Entomology, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Dissertations and Student Research in Entomology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. VIRAL PREVALENCE AMONG SOCIAL BEES IN DIFFERENT LANDSCAPES by Tugce Karacoban A THESIS Presented to the Faculty of Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science Major: Entomology Under the Supervision of Professor Judy Wu-Smart Lincoln, Nebraska December, 2018 VIRAL PREVALENCE AMONG SOCIAL BEES IN DIFFERENT LANDSCAPES Tugce Karacoban, M.S. University of Nebraska, 2018 Advisor: Judy Wu-Smart Honey bees and wild bees provide important pollination services to numerous crops and native plants. In recent years, declines in bee populations have highlighted the importance of the ecological services they provide and the need for more research into the reasons for their decline. Currently, many conservation efforts to mitigate bee losses include increasing forage and habitat, however, there is growing concern over the role interspecific pathogen transmission plays in bee decline. Viruses commonly found in honey bees may be transmitted and pose a threat to other bee species when bees come together at foraging sites. To elucidate the impact of viruses in bee health decline, I examined the roles flowers, bee management, land type, and foraging activity play in viral prevalence. Bees, pollen (collected from foraging bees), in-hive pollen stores, flowers, and other insects on flowers were analyzed for the presence of four common honey bee viruses using RT-PCR sequencing techniques. To further examine the role bee management and life history traits, such as sociality, may play in the transmission and or persistence of viruses, we compared viral profiles from two species of managed social bees (Apis mellifera, Bombus impatiens) and two species of wild social bees (B. griseocollis, Halictus ligatus). Bees were also collected from different landscapes (urban, agricultural, roadsides and conservation parks) and from short, medium, or long blooming plants to determine how floral traits and land management practices may impact viral profiles among bees. Acknowledgements I have so many people to thank that it’s difficult to know where to start. Some contributed directly and some indirectly, but all helped to make this possible. My path to graduate school may not have been as straightforward as it is for most people, but that is a topic for a whole other book. I did not know I wanted to become a Researcher/Scientist. I like to work hard, to read and to learn new things. I was working in veterinary medicine at an Animal hospital (Veterinary Surgical Orthopedic Center). I have always loved animals so it was a big honor to make life better for animals in need. Even if I was feeling that I was doing the best thing in my life, I could not help but feel that I needed to help animals and people at the same time. It is at this point, the importance of bees in the world captured my attention. So I applied for a grant offered by the Republic of Turkey Ministry to complete a graduate degree abroad for the Apiculture Research Institute of Republic of Turkey Ministry of Food, Agriculture and Livestock. I decided to do my master’s degree in USA and approached Judy Wu-Smart to be one of her graduate students. Although I did not have prior knowledge about bees and beekeeping, I was provided opportunities to learn everything from my advisor and help maintain honey bee colonies. I am very honored and grateful to have had Judy Wu-Smart as my advisor and academic mentor. I cannot find enough words to thank her for all her support, advices, mentorship, friendship, and of course, patience. Judy believed on my potential when I did not even know I had any. And for that I will always thank her. I want to thank the support from members of the Bee Lab. The list may be long, but I would be remiss if I did not mention all of them: Kayla Mollet, Katie Lamke, Surabhi Gupta ii Vakil, Natalia Bjorklund, Dustin Scholl, Sheldon Brummel, Courtney Brummel, Jennifer Mai Albrecht. I would like to express my special thanks to two of them. In every point of my struggle, Kayla Mollet helped and encouraged me. Thank you Kayla for all your help and good friendship. You are the BEST! Another special thank you to Katie Lamke, who always had time to help with identification of my bees and made time to answer all of my questions. I would also like to extend my sincere gratitude to my committee members, Drs. Troy Anderson and Sydney Everhart. I could not have had a better group of faculty. Thank you for all your support, invaluable advice and contributions to my research. My family, a big thank you for the countless words of support. It wasn’t easy, but without them it would have been much harder. Thanks to Hayriye Karacoban, Orhan Karacoban, Tolga Karacoban, Zeynep Karacoban and Sema Olgun, my family in Turkey. My partner, best friend, and confidant, Turugsan Olgun. He is my strong shoulder. There are not enough words for me to properly thank you. You made me feel like you were always near me even if we have thousands of miles between us. Thank you so much for your unconditional love. And last, but never least, my mom and dad. I always felt their love just with a phone call or just a smile at the back of a computer screen. They taught me that there is no limitation to show your love and support. They will always be my angels who just want my favor. iii Table of Contents Acknowledgements .............................................................................................................. i List of Figures & Tables .................................................................................................... vi Chapter 1. Literature review 1.1 The importance of bees ..................................................................................................1 1.2 Status of bee decline and contributing stress factors .....................................................3 1.3 Honey bee stressors ........................................................................................................3 1.4 Viruses in Apis bees, other Hymenopterans, and non-Hymenopteran arthropods ........5 1.5 Pathogen transmission ...................................................................................................7 1.6 Pathogen transmission in honey bees ............................................................................9 References cited ................................................................................................................15 Chapter 2. An examination of viruses in four different species of bees collected from agricultural, urban, and natural landscapes 2.1 Abstract ........................................................................................................................19 2.2 Introduction ..................................................................................................................20 2.3 Materials & Methods ...................................................................................................24 2.3.1 Target bee species and site selection .....................................................................24 2.3.2 Collection method of bees .....................................................................................25 2.3.3 Honey bee colonies and apiaries ............................................................................26 2.3.4 Bee tissue dissection ..............................................................................................27 2.3.5 Sample collection of pollen ...................................................................................27 2.3.6 Total RNA extraction from bees and pollen ..........................................................28 2.3.7 RT-PCR method ....................................................................................................29 iv 2.3.8 Target virus selection and assessment ...................................................................29 2.3.9 Statistical analysis ..................................................................................................31 2.3.9.1 Proportion of infected bees across different landscapes ...................................31 2.3.9.2 Distance-based redundancy analysis of virus type ...........................................31 2.3.9.3 Mixed virus infections in honey bee colonies ...................................................32 2.4 Result ...........................................................................................................................32 2.4.1 Virus prevalence in foraging bees collected from different landscapes ................32 2.4.2 Dissimilarities
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