@phdthesis{Hoecherl2015, author = {H{\"o}cherl, Nicole}, title = {Nesting behaviour of the paper wasp Polistes dominula - with special focus on thermoregulatory mechanisms}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-132681}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2015}, abstract = {Wasps of the genus Polistes comprise over 200 species and are nearly cosmopolitan. They show a lack of physiological caste differentiation and are therefore considered as primitively eusocial. Furthermore, paper wasps are placed between the solitary living Eumenidae and the highly social organized Vespinae. Hence, they are often called a "key genus" for understanding the evolution of sociality. Particularly, Polistes dominula, with its small easy manageable nests and its frequent occurrence and wide distribution range is often the subject of studies. In Europe, the invasion of this species into northern regions is on the rise. Since little was known about the nesting behaviour of P. dominula in Central Europe, the basic principles about nesting were investigated in W{\"u}rzburg, Germany (latitude 49°) by conducting a comprehensive field-study spanning three consecutive years. Furthermore, the thermoregulation of individual wasps in their natural habitat had not yet been investigated in detail. Therefore, their ability to respond to external hazards with elevated thorax temperatures was tested. In addition, different types of nest thermoregulation were investigated using modern methods such as infrared thermography and temperature data logger. In the present work, the investigation of basic nesting principles revealed that foundress groups (1-4 foundresses) and nests are smaller and that the nesting season is shorter in the W{\"u}rzburg area than in other regions. The mean size of newly founded nests was 83 cells and the average nesting season was around 4.6 months. The queens neither preferred single (54\%) nor multiple founding (46\%) in this study. The major benefit of multiple founding is an increased rate of survival. During the three years of observation, only 47\% of single-foundress colonies survived, whereas 100\% of colonies that were built by more than two queens, survived. However, an influence of the number of foundresses on the productivity of colonies in terms of number of cells and pupae per nest has not shown up. However, the length of the nesting season as well as the nest sizes varied strongly depending on the climatic conditions of the preceding winter during the three consecutive years. In order to investigate the thermoregulatory mechanisms of individual adult P. dominula wasps, I presented artificial threats by applying smoke or carbon dioxide simulating fire and predator attacks, respectively, and monitored the thorax temperature of wasps on the nest using infrared thermography. The results clearly revealed that P. dominula workers recognized smoke and CO2 and reacted almost instantaneously and simultaneously with an increase of their thorax temperature. The maximal thorax temperature was reached about 65 s after the application of both stressors, but subsequently the wasps showed a different behaviour pattern. They responded to a longer application of smoke with moving to the exit and fled, whereas in case of CO2 the wasps started flying and circling the nest without trying to escape. No rise of the thorax temperature was detectable after an air blast was applied or in wasps resting on the nest. Additionally, the thorax temperatures of queens were investigated during dominance battles. I found that the thorax temperature of the dominant queens rose up to 5°C compared to that of subordinate queens that attacked the former. The study of active mechanisms for nest thermoregulation revealed no brood incubation or clustering behaviour of P. dominula. Furthermore, I found out that wing fanning for cooling the nest was almost undetectable (4 documented cases). However, I could convincingly record that water evaporation is most effective for nest cooling. By the direct comparison of active (with brood and adults) and non-active (without brood and adults) nests, the start of cooling by water evaporation was detected above maximum outside temperatures of 25°C or at nest temperatures above 35°C. The powerful role of water in nest cooling was manifested by an average decrease of temperature of a single cell of about 8°C and a mean duration of 7 min until the cell reached again its initial temperature. The investigation of passive thermoregulatory mechanisms revealed that the nest site choice as well as nest orientation appears to be essential for P. dominula wasps. Furthermore, I was able to show that the architecture of the nests plays an important role. Based on the presented results, it can be assumed that the vertical orientation of cells helps maintaining the warmth of nests during the night, whereas the pedicel assists in cooling the nest during the day.}, subject = {Franz{\"o}sische Feldwespe}, language = {en} } @phdthesis{Hartmann2014, author = {Hartmann, Michael}, title = {Charakterisierung inaktivierender posttranslationaler Modifikationen des GC-A-Rezeptors f{\"u}r das atriale natriuretische Peptid (ANP)}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-97959}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2014}, abstract = {Das atriale natriuretische Peptid (ANP) wird infolge einer Zunahme des atrialen Drucks aus den Myozyten des Atriums sezerniert. Es spielt lokal eine bedeutende, protektive Rolle und wirkt der Entstehung von Herzhypertrophie und Fibrose entgegen. Dar{\"u}ber hinaus kommt ANP vor allem eine wichtige Rolle als endokrines Hormon zu, das den arteriellen Blutdruck und das Blutvolumen regelt. Diese physiologischen Effekte vermittelt das Herzhormon durch seinen Rezeptor, das Transmembranprotein Guanylatzyklase A (GC-A). Durch Bindung von ANP an die extrazellul{\"a}re Dom{\"a}ne der GC-A wird intrazellul{\"a}r, durch die katalytische Dom{\"a}ne des Rezeptors, der sekund{\"a}re Botenstoff cGMP gebildet. Patienten mit einer, durch Bluthochdruck verursachten Herzhypertrophie und Herzinsuffizienz weisen erh{\"o}hte ANP-Konzentrationen im Plasma auf. Die durch ANP vermittelten, protektiven Effekte sind allerdings vermindert. Zahlreiche Studien haben in vitro gezeigt, dass die chronische Inkubation der GC-A mit ihrem Liganden, sowie die Behandlung von GC-A exprimierenden Zellen mit Hormonen wie Angiotensin II, zur Desensitisierung des Rezeptors f{\"u}hren. Der Verlust der Funktionsf{\"a}higkeit geht einher mit der Dephosphorylierung des Rezeptors an spezifischen, intrazellul{\"a}r lokalisierten Aminos{\"a}uren. Durch die Erforschung dieses Mechanismus und Identifizierung m{\"o}glicher Interaktionspartner in vivo k{\"o}nnte der Grundstein f{\"u}r neue oder verbesserte Therapieformen gelegt werden. Im ersten Teil der vorliegenden Arbeit wurde eine k{\"u}rzlich identifizierte Isoform des GC-A-Rezeptors identifiziert, die durch alternatives Spleißen des Exons 4 entsteht und in einer Vielzahl untersuchter Gewebe der Maus vorkommt. Die Deletion umfasst 51 Basenpaare und resultiert in einem um 17 Aminos{\"a}uren verk{\"u}rzten GC-A-Rezeptor (GC-AΔLys314-Gln330). Molekulare Modellierungen der extrazellul{\"a}ren Dom{\"a}nen des wildtypischen GC-A-Rezeptors und der Isoform zeigten, dass sich die Deletion im membrannahen Bereich der extrazellul{\"a}ren Dom{\"a}ne und damit deutlich entfernt von der ANP-Bindungsdom{\"a}ne befindet. Oberfl{\"a}chenbiotinylierungs- und Zellfraktionierungsversuche zeigten, dass die Isoform des GC-A-Rezeptors an der Oberfl{\"a}che von Zellmembranen transient transfizierter HEK 293-Zellen pr{\"a}sentiert wird. Jedoch zeigten die ANP-Stimulationsexperimente unter Anwendung von cGMP-Radioimmunassay (cGMP-RIA) und F{\"o}rster-Resonanzenergietransfer (FRET)-Messungen, dass die Isoform nicht zur ANP-vermittelten intrazellul{\"a}ren cGMP-Bildung stimuliert werden kann. Im Rahmen von ANP-Bindungsstudien mit 125I-ANP wurde gezeigt, dass GC-AΔLys314-Gln330 die F{\"a}higkeit zur Bindung des Liganden ANP verloren hat. Jedoch zeigten die Koimmunpr{\"a}zipitationsversuche, dass die Isoform des GC-A-Rezeptors Heterodimere mit dem wildtypischen GC-A-Rezeptor bilden und dadurch die ligandeninduzierte Bildung von cGMP reduzieren kann. In vivo konnte gezeigt werden, dass unter Angiotensin II-induzierter Hypertonie die mRNA-Expression f{\"u}r GC-AΔLys314-Gln330 in der Lunge gesteigert, und gleichzeitig die ANP-vermittelte cGMP-Bildung deutlich reduziert ist. Daher kann davon ausgegangen werden, dass das alternative Spleißen ein regulierender Mechanismus ist, der auf den ANP/GC-A-Signalweg Einfluss nimmt. Angiotensin II-induziertes alternatives Spleißen des GC-A-Gens kann daher einen neuen Mechanismus f{\"u}r die Verringerung der Sensitivit{\"a}t des GC-A-Rezeptors gegen{\"u}ber ANP darstellen. Im zweiten Teil der vorliegenden Arbeit wurden transgene Tiere mit kardiomyozytenspezifischer {\"U}berexpression eines Epitop-getaggten GC-A-Rezeptors generiert. Durch dieses Modell sollte es erm{\"o}glicht werden, den Rezeptor aus murinem Gewebe anreichern und aufreinigen zu k{\"o}nnen um danach Analysen zu posttranslationalen Ver{\"a}nderungen und m{\"o}glichen Interaktionspartnern durchzuf{\"u}hren. Zun{\"a}chst wurde in eine FLAG-Epitop-getaggte GC-A zus{\"a}tzlich ein HA-tag, sowie eine Erkennungssequenz f{\"u}r die Protease des tobacco etch virus (TEV) eingef{\"u}gt. Die Expression und Funktionsf{\"a}higkeit des modifizierten Rezeptors wurde durch ANP-Stimulationsexperimente unter Anwendung von cGMP-RIA und FRET-Messungen verifiziert. Die Funktionsf{\"a}higkeit der TEV-Erkennungssequenz wurde durch die Elution mittels TEV-Protease nach Immunpr{\"a}zipitation (IP) nachgewiesen. In vivo wurde an M{\"a}usen die Expression und Lokalisation der GC-A auf Proteinebene, unter Anwendung von Zellfraktionierungsexperimenten und Immunpr{\"a}zipitationen, {\"u}berpr{\"u}ft. Die entstandenen transgenen Tiere zeigten eine deutliche, in den Zellmembranen von Kardiomyozyten lokalisierte, {\"U}berexpression des Rezeptors. Dieser konnte {\"u}ber das HA-tag angereichert und aufgereinigt werden. Um die Funktionsf{\"a}higkeit des modifizierten Rezeptors in vivo nachzuweisen, wurde in zwei Versuchsreihen kardiale Hypertrophie durch chronische Applikation von Angiotensin II induziert. Es wurde postuliert, dass die {\"U}berexpression funktionsf{\"a}higer GC-A im Herzen die Tiere vor Herzhypertrophie sch{\"u}tzt. Die Ergebnisse der Studien zeigen allerdings, dass die generierten transgene Tiere trotz kardiomyozytenspezifischer {\"U}berexpression des Rezeptors nicht den erwarteten Schutz vor Herzhypertrophie aufwiesen, sondern {\"a}hnlich wie ihre wildtypischen Geschwistertiere reagieren. Jedoch gelang es mit Hilfe des {\"U}berexpressionsmodells zusammen mit anderen Mitarbeitern der AG Kuhn eine zuvor in vitro beschriebene Interaktion des GC-A-Rezeptors mit den Kationenkan{\"a}len TRPC3 und TRPC6 in vivo nachzuweisen. Somit besteht die M{\"o}glichkeit die Epitope und das murine {\"U}berexpressionsmodell auch zuk{\"u}nftig zu nutzen, um Interaktionspartner der GC-A zu identifizieren.}, subject = {Guanylatzyklase}, language = {de} } @phdthesis{Rutschmann2023, author = {Rutschmann, Benjamin}, title = {Occurrence and population density of wild-living honey bees in Europe and the impact of different habitat types on their foraging and overwintering success}, doi = {10.25972/OPUS-28673}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-286732}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {The original habitat of native European honey bees (\(Apis\) \(mellifera\)) is forest, but currently there is a lack of data about the occurrence of wild honey bee populations in Europe. Prior to being kept by humans in hives, honey bees nested as wild species in hollow trees in temperate forests. However, in the 20th century, intensification of silviculture and agriculture with accompanying losses of nesting sites and depletion of food resources caused population declines in Europe. When the varroa mite (Varroa destructor), an invasive ectoparasite from Asia, was introduced in the late 1970s, wild honey bees were thought to be eradicated in Europe. Nevertheless, sporadic, mostly anecdotal, reports from ornithologists or forest ecologists indicated that honey bee colonies still occupy European forest areas. In my thesis I hypothesize that near-natural deciduous forests may provide sufficient large networks of nesting sites representing refugia for wild-living honey bees. Using two special search techniques, i.e. the tracking of flight routes of honey bee foragers (the "beelining" method) and the inspection of known cavity trees, I collected for the first time data on the occurrence and density of wild-living honey bees in forest areas in Germany (CHAPTER 3). I found wild-living honey bee colonies in the Hainich national park at low densities in two succeeding years. In another forest region, I checked known habitat trees containing black woodpecker cavities for occupation by wild-living honey bee colonies. It turned out that honey bees regularly use these cavities and occur in similar densities in both studied forest regions, independent of the applied detection method. Extrapolating these densities to all German forest areas, I estimate several thousand wild-living colonies in Germany that potentially interact in different ways with the forest environment. I conclude that honey bees regularly colonize forest areas in Germany and that networks of mapped woodpecker cavities offer unique possibilities to study the ecology of wild-living honey bees over several years. While their population status is ambiguous and the density of colonies low, the fact that honey bees can still be found in forests poses questions about food supply in forest environments. Consequently, I investigated the suitability of woodlands as a honey bee foraging habitat (CHAPTER 4). As their native habitat, forests are assumed to provide important pollen and nectar sources for honey bee colonies. However, resource supply might be spatially and temporally restricted and landscape-scale studies in European forest regions are lacking. Therefore, I set up twelve honey bee colonies in observation hives at locations with varying degree of forest cover. Capitalizing on the unique communication behaviour, the waggle dance, I examined the foraging distances and habitat preferences of honey bees over almost an entire foraging season. Moreover, by connecting this decoded dance information with colony weight recordings, I could draw conclusions about the contribution of the different habitat types to honey yield. Foraging distances generally increased with the amount of forest in the surrounding landscape. Yet, forest cover did not have an effect on colony weight. Compared to expectations based on the proportions of different habitats in the surroundings, colonies foraged more frequently in cropland and grasslands than in deciduous and coniferous forests, especially in late summer when pollen foraging in the forest is most difficult. In contrast, colonies used forests for nectar/honeydew foraging in early summer during times of colony weight gain emphasizing forests as a temporarily significant source of carbohydrates. Importantly, my study shows that the ecological and economic value of managed forest as habitat for honey bees and other wild pollinators can be significantly increased by the continuous provision of floral resources, especially for pollen foraging. The density of these wild-living honey bee colonies and their survival is driven by several factors that vary locally, making it crucial to compare results in different regions. Therefore, I investigated a wild-living honey bee population in Galicia in north-western Spain, where colonies were observed to reside in hollow electric poles (CHAPTER 5). The observed colony density only in these poles was almost twice as high as in German forest areas, suggesting generally more suitable resource conditions for the bees in Galicia. Based on morphometric analyses of their wing venation patterns, I assigned the colonies to the native evolutionary lineage (M-lineage) where the particularly threatened subspecies \(Apis\) \(mellifera\) \(iberiensis\) also belongs to. Averaged over two consecutive years, almost half of the colonies survived winter (23 out of 52). Interestingly, semi-natural areas both increased abundance and subsequent colony survival. Colonies surrounded by more semi-natural habitat (and therefore less intensive cropland) had an elevated overwintering probability, indicating that colonies need a certain amount of semi-natural habitat in the landscape to survive. Due to their ease of access these power poles in Galicia are, ideally suited to assess the population demography of wild-living Galician honey bee colonies through a long-term monitoring. In a nutshell, my thesis indicates that honey bees in Europe always existed in the wild. I performed the first survey of wild-living bee density yet done in Germany and Spain. My thesis identifies the landscape as a major factor that compromises winter survival and reports the first data on overwintering rates of wild-living honey bees in Europe. Besides, I established methods to efficiently detect wild-living honey bees in different habitat. While colonies can be found all over Europe, their survival and viability depend on unpolluted, flower rich habitats. The protection of near-natural habitat and of nesting sites is of paramount importance for the conservation of wild-living honey bees in Europe.  }, subject = {Biene}, language = {en} }