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Ph.D. Dissertation MASARYK UNIVERSITY FACULTY O F S CIENCE DEPARTMENT OF BOTANY AND ZOOLOGY Ph.D. Dissertation BRNO 2017 Msc. Willems Kamila MASARYK UNIVERSITY FACULTY O F S CIENCE DEPARTMENT OF BOTANY AND ZOOLOGY Intraspecific variability and dispersal propensity of the palaearctic bugs of the genus Cimex (Heteroptera: Cimicidae) M.Sc. Willems Kamila Supervisor : doc. Mgr. Tomáš Bartoni čka, Ph.D Consultant : Mgr. Ond řej Balvín, Ph.D Bibliografický záznam Autor: Mgr. Kamila Willems, Přírodov ědecká fakulta, Masarykova Univerzita, Ústav botaniky a zoologie Vnitrodruhová variabilita a schopnost ší ření u palearktických št ěnic rodu Cimex Název práce: (Heteropetra: Cimicidae) Studijní program: Biologie Studijní obor: Zoologie Vedoucí práce: doc. Mgr. Tomáš Bartoni čka, Ph.D Akademický rok: 2017/2018 Po čet stran: 124 Klí čová slova: Ektoparazit, št ěnice, Cimex, k oevoluce Bibliographic Entry Author: M.Sc. Kamila Willems, Faculty of Science, Masaryk University, Department of Botany and Zoology Title of Thesis: Intraspecific variability and dispersal propensity of the palaearctic bugs of the genus Cimex (Heteroptera: Cimicidae) Field of Study Zoology Supervisor: doc. Mgr. Tomáš Bartoni čka, Ph.D Academic Year: 2017/2018 Number of Pages: 124 Ectoparasites, bed bugs, bat bugs, Cimex , Keywords: coevolution ABSTRACT Main research was focus on two haematophagous ectoparasitic species from family Cimicidae. First directly related to human - bed bug Cimex lectularius which expands widely last years and is more and more often found in human dwellings. As ectoparasite that can leave freely and use host only during taking a blood meal, and can starve for a very long period of the time, it is hard to exterminate. Knowledge about its biology, ecology and morphology can be very useful in solving many problems connected with its infestation. Especially that it was found to create associations both with humans and other crucial host, namely bats. Regarding those facts our research focused on investigation of sucking abilities of both bat and human associated bugs to estimate level of association with both hosts. We have found that both human and bat associated bed bugs can suck on non-specific for them host. Further observations of successful survival, reproduction and development abilities shown us clearly that in cross-feeding experiment type of the host, specific/non-specific, seems to have an impact on survival, moulting and development rate in both cross-feedings conducted in vitro. Because bed bugs were never found on some bat species, we investigated blood composition with focus on RBC (red blood cells) size and density. We did not find significant differenced in RBC measurements between specific and non- specific bat host species. Following the fact that both human and bat associated bugs can suck on non- specific host, it was important to see if both lineages in case of meeting are being able to mate, reproduce and produce successfully progeny. Further hybridization experiments between two lineages human and bat associated bed bug ( Cimex lectularius ) from Central Europe (Czech Republic) were conducted under laboratory conditions. Couples mix of both lineages (interspecific mating) were compared with couples from same lineage, same locality and same lineage, different localities (intraspecific mating). In case of mixed couples from different lineages creating hybrids was unsuccessful, no eggs no progeny was produced. Second species, indirectly connected with human and closely associated with bat hosts - C. pipistrelli was investigated in terms of transmission by its host. We do not posses much information about bat bugs sucking on humans, nevertheless in many cases bats and humans share same shelter which increases risk of infestation. Transmission experiments were conducted on 2 most common bat bugs vectors – noctule bat ( Nuctalus noctula ) and greater mouse – eared bat ( Myotis myotis ) as well on non common bat species – daubenton´s bat (Myotis daubentonii ). Results clearly confirmed that bat bugs prefer transmission on bigger bat species such as noctule or greater mouse-eared bat than daubenton´s bat. What is more from both bigger bats they shows clear preferences towards noctule bat which in fact is mostly mist-netted bat which carries bugs on his body. Research proved also that bugs that mainly departed from bat box on bats body were mated females not virgins, as a second one has no possibility to start new infestation. Following bat bug transmission possibilities, we conducted genetic research at local and regional scale in Central Europe. Developed polymorphic loci has shown homogenous character in populations of bat bugs from localities across Czech Republic, Slovakia and Ukraine. © Kamila Willems, Masarykova Univerzita, 2017 Acknowledgements I would like to express my special appreciation and thanks to my advisor and supervisor, doc. Mgr. Tomáš Bartoni čka, Ph.D who has a tremendous mentor and help for me. I would like to thank him for encouraging me in my research and for allowing me to grow as a research scientist. I would also like to thank also my consultant Mgr. Ond řej Balvín, Ph.D, doc. Mgr. et Mgr. Josef Bryja, Ph.D and Mgr. Adam Kone čný, Ph.D. for all advices and sharing with me their experience and knowledge. And to all the other people that made my PhD time at Masaryk University unforgettable experience. A special thanks to my family. Words cannot express how grateful I am to my mother and sister for all of the sacrifices that you’ve made on my behalf. Your words of support, encouraging and your patience. I would also like to thank all of my friends all over the Europe, who supported me in writing, and incented me to strive towards my goal. At the end I would like to express appreciation to my beloved husband Wout Willems who spent sleepless nights with me and was always my support in the hardest moments. Prohlášení Prohlašuji, že jsem svoji diserta ční práci vypracovala samostatn ě s využitím informa čních zdroj ů, které jsou v práci citovány. Brno ……………………………… Kamila Willems 1. Introduction .................................................................................................................................................... 2 1.1 Host specificity ...................................................................................................................................... 2 1.2 Reproductive isolation in insects – inter- and intraspecific matings..................................................... 5 1.3 Host transmission and genetic structure of bat ectoparasites............................................................ 11 1.4 Traumatic insemination as mating strategy ........................................................................................ 14 2. Model organisms........................................................................................................................................... 16 2.1 Bed bug – Cimex lectularius ................................................................................................................ 16 2.2 Bat bug – Cimex pipistrelli................................................................................................................... 17 2.3 Human and bats as bed bug hosts....................................................................................................... 18 2.4 Bat hosts.............................................................................................................................................. 20 2.5 Some hematological aspects of hosts – specific versus non-specific bat host.................................... 22 3. Main aims of the thesis................................................................................................................................. 26 4. Summary of main results .............................................................................................................................. 28 4.1 Two different lineages of bed bug (Cimex lectularius) reflected in host specificity............................ 29 4.2 Hybridization experiments on bed bugs from two lineages (bat and human associated).................. 31 4.3 Erythrocyte size as potential explanation of host specificity in bed bugs........................................... 33 4.4 Development of multiplex panels of polymorphic microsatellite loci for bat bug (Cimex pipistrelli): a pilot methodological study. .............................................................................................................................. 34 4.4.1 Abstract........................................................................................................................................... 34 4.4.2 Microsatelites as genetic tools in insects genetic ........................................................................... 35 4.4.3 Co-evolution of host and ectoparasite genetic variability .............................................................. 37 4.4.4 Material and methods..................................................................................................................... 40 4.4.5 Results............................................................................................................................................. 42 4.4.6 Discussion........................................................................................................................................ 44 4.5 Bat bugs (Cimex pipistrelli) transmission propensity in three bat species.........................................
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