Made available in DSpace on 2014-07-29T15:13:38Z (GMT). No. of bitstreams: 1 Tese_Kennedy_2010.pdf: 1273886 bytes, checksum: d53179ca59a4af096e34e7c640527b2d (MD5) Previous issue date: 2010-08-13 / Amblyomma cajennense, commonly known as Cayenne tick, is a three-host tick widely distributed in Brazil and in the rest of Latin American countries. Owing to its low host specificity, it plays an important role in the transmission of pathogens in animals and man. Chemical communication is a vital component in tick biology whereby pheromones are largely responsible in the regulation of mating, recognition of sexual partner and copulation. By understanding the behaviour of A. cajennense and the pheromones involved, there exists practical possibility for the use of these substances as an additional method of control thereby reducing the use of acaricides. Based on this, the aims of this work were to quantify and elucidate behaviours induced by pheromones emitted by females A. cajennense as well as identify and characterize qualitatively and biologically the pheromone produced by males. Engorged females were collected from naturally-infested horses for the establishment of laboratory colonies for use in this study. The pheromones were obtained by extraction in hexane and sonication by ultra-sound of fed males and females. The female sex pheromone, 2,6-dichlorophenol (2,6-DCP), was quantified by GC/MS using internal standard and standard addition methods through construction of calibration curves using 5-bromine-4-hydroxy-3-methoxybenzaldehyde and 2,6-DCP, respectively. The male extract was analysed chemically by GC/MS in comparison with standards. As concerns biological tests, glass beads were treated with two femaleequivalents of the female pheromone and 2,6-DCP in an equivalent concentration and exposed to males to observe the induced behaviour(s). Attraction to the male pheromone was evaluated by release of individual unfed, virgin females to three concentrations (1, 5 and 10 male-equivalents) of the extract in olfactometer bioassays. Orientations towards the odour source were analysed by Kruskal-Wallis and displacement angles by Circular Statistics. Aggregation was evaluated by releasing the tick stages in Petri dish bioassays shortly followed by introduction of filter paper strips impregnated with 10 male-equivalents of the extract and recording tick responses on camera. Data analysis was by Chi-square test. Tests for attachment were carried out in vivo in rabbits, Oryctolagus cuniculus, and the released ticks observed in determined intervals between 20min and 24hrs. The data was analysed by Wilcoxon signed-ranked test and Friedman s repeated measures ANOVA. The results indicated that each female emits 2.15 ng ml-1 of pheromone on average. The male extract was positive for benzoic acid, nonanoic acid, salicylic acid, benzaldehyde, 2,6-DCP, limonene and methyl salicylate. The female extract and 2,6- DCP stimulated mounting in up to 80% and 73% of all cases, respectively, confirming that 2,6-DCP alone mediates mounting behaviour in A. cajennense. There was no statistically significant attraction or aggregation in males or females induced by the male pheromone. However, significantly faster and greater attachment was recorded in both. The behavioural responses corresponded with the pheromone s composition. It was concluded that the male pheromone promotes neither attraction nor aggregation but is responsible for faster fixation of A. cajennense adults. The pheromone also contains different proportions and differs from other Amblyomma species as far as the presence or absence of certain components is concerned. The results serve as a first step in advanced studies aimed at development of novel strategies for the management of this species using these pheromones. / Amblyomma cajennense, vulgarmente conhecido como carrapato-estrela, é um carrapato trioxeno que está amplamente difundido no território brasileiro e nos demais países da América Latina. Dada a sua baixa especificidade, assume um importante papel na transmissão de patógenos entre os animais e o homem. A comunicação química é um componente vital na biologia dos carrapatos, onde feromônios são majoritariamente responsáveis na regulação da busca pelo parceiro e a cópula. Compreendendo o comportamento de A. cajennense e os feromônios envolvidos, existem possibilidades práticas do uso destas substâncias como uma forma alternativa e satisfatória de controlálos minimizando o uso de acaricidas. Diante disso, procurou-se neste trabalho, quantificar e elucidar comportamentos instigados por feromônio produzido por fêmeas bem como identificar e caracterizar qualitativa e biologicamente feromônio de machos. Fêmeas ingurgitadas de A. cajennense foram colhidas de eqüinos naturalmente infestados para estabelecimento de colônias no laboratório. Os feromônios foram obtidos por extração e sonicação em hexano de machos ou fêmeas de carrapatos alimentados. Quantificou-se o feromônio sexual, o 2,6-diclorofenol (2,6-DCF), em fêmeas pelas técnicas de padrão interno, com 5-bromo-4-hidróxi-3-metóxi-benzaldeido como referência interna, e adição padrão por meio de cromatografia gasosa acoplada à espectrometria de massas (CG/EM) através de construção de curvas de calibração. O extrato dos machos foi analisado quimicamente por CG/EM diante de padrões sintéticos. No que concerne aos testes biológicos, pérolas de vidro foram tratadas com dois equivalentes-fêmea de amostra de feromônio extraído de fêmeas e com 2,6-DCF em uma dose equivalente e expostas aos machos para observar comportamento(s) provocado(s). A atração diante do feromônio de machos foi avaliada pela liberação individual de fêmeas, machos e ninfas virgens, não alimentadas, frente a três concentrações (1, 5 e 10 equivalentes-macho) do extrato de machos, em um olfatômetro. As orientações à fonte de odor foram analisadas por teste de Kruskal-Wallis e os ângulos por Estatística Circular. A agregação foi avaliada pela aplicação de 10 equivalentes-macho do extrato em arenas de placas de Petri e os dados analisados por Chi-quadrado. Testes de fixação foram feitos in vivo em coelhos, Oryctolagus cuniculus, onde os carrapatos foram liberados e observados em intervalos determinados entre 20min e 24hs e suas respostas analisadas por testes de Wilcoxon e ANOVA Friedman. Os resultados mostraram que uma fêmea libera em média 2,15 ng mL-1 de feromônio. O extrato de machos apresentou ácido benzóico, ácido nonanóico, ácido salicílico, benzaldeido, 2,6-DCF, limoneno e salicilato de metila. O extrato de fêmeas e o 2,6-DCF sintético estimularam a monta em até 73% de casos, respectivamente, confirmando que o composto é responsável por mediar à monta em A. cajennense. Não houve atração nem agregação estatisticamente maior dos machos e fêmeas diante do feromônio procedente de machos. Entretanto, uma fixação significativamente maior e mais rápida foi registrada dentro de 24 h em ambos os sexos. Os achados de respostas biológicas corresponderam com a composição química de feromônio de machos. Concluiu-se que o feromônio de machos não promove atração nem agregação, mas é responsável pela fixação de adultos de A. cajennense. O feromônio também possui diferenças na proporção e na presença de determinados compostos descritos em outras espécies do gênero Amblyomma. Os resultados estão discutidos como um ponto de partida para estudos avançados visando o desenvolvimento de novas estratégias de controle e de manejo desta espécie utilizando esses feromônios.

A critical mechanism underpinning current biological diversity is the extent to which one species mates with, or avoids mating with, another. However, little is known about the factors that mediate hybridisation, especially during the initial and rarely observed stages of secondary contact when interspecific interactions have not responded to selection. In particular, whilst hybridisation is ultimately a behavioural phenomenon, the role of behaviour in mediating hybridisation and how it is influenced by environmental and circumstantial factors is rarely investigated. Recently introduced species provide us with unequalled opportunities to study these factors. In this thesis I examine the role of behavioural mechanisms, in particular male-male competition and mate choice, in mediating mating patterns between two genetically and phenotypically distinct lineages of European wall lizard (Podarcis muralis) that have come into recent secondary contact through human introductions. In Chapter Two, I investigated how sexual selection during allopatry is responsible for creating stark differences in phenotypic traits such as body size and weapon performance evident in the two lineages today, ultimately explaining the strong biases in dominance during territorial disputes between males. However, I also show that even given this asymmetry in male competitive ability, the extent to which it extrapolates into greater access to females in naturalistic, outdoor enclosures depends strongly on the spatial clustering of basking sites, a critically important resource for many ectotherms. In contrast to initial predictions suggested by asymmetries in male competition outlined in the previous chapter, in Chapter Three I show that both paternity and courtship behaviour was strongly assortative in the outdoor enclosures. Further investigation through staged experiments on olfactory mate choice, mating trials and analyses on specific behavioural data obtained in an enclosure experiment, I show that lineage based dominance actually contributes to assortative mating patterns in conjunction with weak conspecific male choice. In contrast, female choice seems to play no role in mediating the mating patterns observed between the two lineages. In Chapter Four I had the rare opportunity to examine the morphological and behavioural factors that predict why animals should hybridise in the first place, using the data obtained in the enclosure experiment above. I found that hybridisation was particularly common between small individuals of the larger lineage and large individuals of the smaller lineage; a result that corroborates the mechanisms determining the assortative patterns uncovered in Chapter Three. Additionally, hybridisation rates were particularly high in less dominant individuals, which I suggest is due to subordinate males having reduced opportunities for courting conspecific females due to male-male competition, requiring them to become less ‘choosy’ and therefore more likely to mate with heterospecifics. Finally, secondary contact cannot occur without at least one lineage coming into a new environment, and yet relatively little attention is paid to how this environmental change can affect the signals involved in intraspecific communication and mate choice. In Chapter Five I show that a change in the amount of time male lizards spend thermoregulating (a likely consequence of arriving in a new environment) significantly changes the chemical composition of their scent marks. However, whilst female lizards were able to detect these effects, they did not seem to base their mating decisions on them. Nevertheless, this result raises interesting questions about the potential function and consequences of this plasticity, and highlights the importance of considering plasticity in chemical communication in heterogeneous environments. Overall, this thesis shows the critically important role of behaviour in mediating intra- and interspecific mating patterns during recent secondary contact. In particular, it highlights how the direction and extent of hybridisation and competition are influenced by the degree to which differing morphological and behavioural phenotypes interact over a heterogeneous environment, particularly during the initial stage of secondary contact when mate choice has not had the chance to respond to the selective pressures of hybridisation.

Eine sichere Bindung zwischen Mutter und Kind in den ersten Lebensjahren ist prägend für die Entwicklung eines Kindes. Die Qualität dieser Bindung ist ein wichtiger Prädiktor für langfristige physische und psychische Gesundheit. Für den Aufbau einer starken Bindung ist die Investition von Ressourcen seitens der Fürsorgeperson auf zeitlicher, physischer und emotionaler Ebene notwendig. Multimodale biologische Hinweisreize seitens des Kindes fördern dieses Engagement. Zunächst dienen solche Signale der Identifikation des eigenen Nachwuchses (kin recognition), um nachfolgend gezielt Ressourcen zu investieren. Darüber hinaus können infantile Stimuli affektive Reaktionen vermitteln, die den Bindungsaufbau erleichtern. In diesem Zusammenhang sind auch olfaktorische Signale, z. B. Körpergerüche, wirksam, bislang gibt es jedoch nur wenig systematische Forschung zu ihrem Einfluss auf die Eltern-Kind-Beziehung. Einzelne Studien zeigen, dass Mütter ihre Kinder am Geruch erkennen können und dass kindliche Körpergerüche auch auf neuronaler Ebene positive Reaktionen vermitteln, wobei jedoch unklar ist, wie spezifisch die neuronale Aktivität für den Geruch des eigenen Kindes ist. In der vorliegenden Arbeit soll der Einfluss von kindlichen Körpergerüchen in der Mutter-Kind- Beziehung über die kindliche Entwicklungsspanne unter Berücksichtigung genetischer, hormoneller und neurobiologischer Faktoren untersucht werden. In Veröffentlichung 1 wurde geprüft, ob Mütter ihre Kinder am Geruch identifizieren können, ob sie diesen präferieren und wie beides mit genetischen und hormonellen Faktoren sowie dem kindlichen Entwicklungsstatus interagiert. Dafür wurden N = 164 Müttern mit ihren biologischen Kindern (N = 226 Kinder zwischen 0 und 18 Jahren) in die Studie eingeschlossen. Die Mütter bewerteten die Körpergerüche des eigenen und fremder Kinder, die sich im Entwicklungsstatus sowie der genetischen Ähnlichkeit unterschieden. Die genetische Ähnlichkeit wurde über das Humane Leukozytenantigen(HLA)-Profil abgebildet, der Entwicklungsstatus wurde anhand der Steroidhormonkonzentration (Testosteron, Estradiol) und einer standardisierten Einschätzung des pubertären Status erfasst. Es zeigte sich, dass die Mütter den Geruch ihres eigenen Kindes über dem Zufallsniveau identifizieren konnten und diesen Geruch präferierten. Dies galt für alle Altersgruppen, mit Ausnahme der frühen Pubertät. In diesem Alter (9-13 Jahre) konnten die Mütter den Geruch ihres Kindes weder identifizieren, noch bevorzugten sie ihn im Vergleich zu fremden Körpergerüchen. Bei den eigenen Söhnen war die Abnahme der Präferenz mit dem Anstieg des Testosteronlevels assoziiert. Mit zunehmendem Alter des Kindes (14-18 Jahre) ähnelte das Bewertungsverhalten der Mütter wieder dem vor Pubertätsbeginn, was vermuten lässt, dass die Mütter sich in diesem Zeitraum an den veränderten Geruch des Kindes gewöhnen und somit die Vertrautheit des Geruchs eine wichtige Rolle für die Wahrnehmung spielt. Zusätzlich legen die Ergebnisse nahe, dass genetische Ähnlichkeit über Körpergerüche transportiert wird: Der Geruch des eigenen Kindes wurde zwar global bevorzugt, im paarweisen Vergleich zeigte sich jedoch, dass sich die Bewertung für den Geruch des eigenen Kindes nicht signifikant von der Bewertung des gleichaltrigen und HLA-ähnlichen Kindes unterschied. Dies lässt darauf schließen, dass sich genetische Ähnlichkeit positiv auf die Geruchsbewertung im Kontext der Eltern-Kind-Bindung auswirkt. Für die zweite Veröffentlichung wurde anhand derselben Stichprobe getestet, ob Mütter den Entwicklungsstatus des Kindes anhand von Körpergerüchen klassifizieren können und welche Prädiktoren für die Klassifikation entscheidend sind. Dafür wurden sie gebeten, die jeweilige Altersgruppe des Kindes einzuschätzen, von dem der Geruch stammte. Die Ergebnisse demonstrieren, dass Mütter den kindlichen Entwicklungsstatus (prä- bzw. postpubertär) mit einer Genauigkeit von 64 % detektieren können und insgesamt dazu tendieren, kindliche Körpergerüche als präpubertär zu klassifizieren. Die mütterliche Klassifikationsleistung war besser, wenn die Probandinnen Geruchsproben aus der gleichen Altersgruppe wie der des eigenen Kindes beurteilten. Die subjektive Bewertung der Proben hinsichtlich Angenehmheit und Intensität sowie die Einschätzung des pubertären Status waren signifikante Prädiktoren für die entwicklungsbedingte Klassifikation eines Geruchs, während sich der Steroidhormonstatus des Kindes nicht auf die mütterliche Einschätzung auswirkte. Die dritte Veröffentlichung dieser Doktorarbeit diente als methodische Pilotstudie für die spezifische Untersuchung des Einflusses von Babygerüchen auf die neuronale Verarbeitung im mütterlichen Gehirn. Aus anderen Modalitäten ist bekannt, dass kindliche Stimuli Niedlichkeit vermitteln, welche mit belohnungsspezifischer neuronaler Aktivität einhergeht. Dies ist für Babygerüche bisher jedoch kaum erforscht. Die Präsentation von Körpergerüchen zur Ableitung neuronaler Korrelate im Rahmen von funktioneller Magnetresonanztomographie (fMRT) ist aufgrund von Stimuluseigenschaften sowie methodischen Schwierigkeiten herausfordernd. Bislang existieren nur wenige Studien zur neuronalen Verarbeitung von Körpergerüchen ohne einheitliche Konvention über eine geeignete Stimuluspräsentation. Im Rahmen dieser Doktorarbeit sollte daher ein effizientes Design entwickelt werden, welches neuronale Aktivität in Reaktion auf Babygerüche optimal abbildet. Dafür wurden zwei Stimuluspräsentationen verglichen, die sich in Art, Dauer und Frequenz unterschieden. Die kurze, kontinuierliche Reizdarbietung rief im Vergleich zu einer langen, gepulsten Präsentation global stärkere Aktivierungen hervor, weshalb diese als Design empfehlenswert ist, um robuste neuronale Korrelate zu erhalten. Allerdings zeigten sich differentielle Effekte in Abhängigkeit der Hirnregionen, weshalb je nach interessierendem Areal spezifisch zwischen Länge, Dauer und Art der Stimuluspräsentation abgewogen werden sollte. Das kurze Präsentationsdesign wurde im Rahmen der weiterführenden fMRT-Studie verwendet. Diese veranschaulichte, dass Babygerüche Belohnungsareale sowie Netzwerke aktivieren, die Angehmheit, Niedlichkeit und Motivation zur Fürsorge (Pleasure-Netzwerk) kodieren. Die Aktivierungsstärke des Netzwerks sagte dabei vorher, wie angenehm die Mütter den Geruch des eigenen Babys bewerteten. Im Gegensatz zu den Verhaltensdaten aus Veröffentlichung 1, in denen sich eine klare Präferenz für das eigene Kind zeigte, konnte kein Unterschied zwischen der neuronalen Reaktion auf den Geruch des eigenen im Vergleich zu einem fremden Baby gefunden werden. Daher gilt es, die Universalität des Babygeruchs als einen Stimulus, der Niedlichkeit vermittelt, in nachfolgenden Studien systematisch zu überprüfen. Zusammenfassend stellt diese Arbeit dar, dass kindliche Körpergerüche als Chemosignale in der Mutter-Kind-Beziehung wirken und sowohl zur Identifikation des eigenen Kindes beitragen als auch affektive Komponenten vermitteln. Außerdem wurde herausgefunden, dass Körpergerüche Informationen über genetische Ähnlichkeit und den Entwicklungsstatus des Kindes transportieren. Es bleibt offen, welche Faktoren auf molekularer Ebene tatsächlich die Veränderung des Körpergeruchs ausmachen. Chemosensorische Profilanalysen können in zukünftigen Untersuchungen Aufschluss darüber geben. Darüber hinaus sind Langzeitstudien notwendig, um die hier dargestellten assoziativen Zusammenhänge auch über den individuellen Entwicklungsverlauf abzubilden und somit Mechanismen der olfaktorisch vermittelten Eltern-Kind-Beziehung ableiten zu können. Langfristig sollen diese Informationen dazu beitragen, Strategien zur Förderung der Eltern-Kind-Beziehung zu generieren und bisher bestehende Interventionen (wie z. B. Neurofeedbacktraining) auf olfaktorische Stimuli auszuweiten.:Inhaltsverzeichnis Danksagung 4 1 Zusammenfassung 6 2 Summary 9 3 Einführung in die Thematik 12 4 Studienziele: Abgeleitete Forschungsfragen und Hypothesen 21 5 Methodik der Untersuchungen 23 6 Zusammenfassung der Ergebnisse 26 7 Diskussion und Ausblick 29 8 Literaturverzeichnis 40 Anhang I. Verzeichnis der wissenschaftlichen Veröffentlichungen, Konferenzbeiträge und andere Leistungen A Veröffentlichungen der Dissertation und dazugehörige Angaben B Weitere Veröffentlichungen während der Promotionsphase C Konferenzbeiträge und andere Leistungen während der Promotionsphase II. Letters of Acceptance III. Erklärung zur Eröffnung des Promotionsverfahrens IV. Bestätigung der Einhaltung der folgenden aktuellen gesetzlichen Vorgaben / A secure bond between mother and child in the first years of life is crucial for the development of a child. The quality of this bond is an important predictor of long-term physical and mental health. To create such a bond, the caregiving person has to invest resources at a temporal, physical and emotional level. Multimodal biological infantile cues facilitate this commitment. Initially, such signals serve to identify one's own offspring (kin recognition) in order to invest resources in a targeted manner. In addition, infantile stimuli can mediate affective reactions that support bonding. In this context, olfactory signals, e.g. body odors, are also effective, but so far there is little systematic research on their influence on the parent-child relationship. Individual studies show that mothers can recognize their children by their body odor and that infantile body odors also mediate positive reactions at the neural level, although it is unclear how specific they are for their own child. The present study investigates the influence of children ́s body odors in the mother-child relationship over the developmental span, integrating genetic, hormonal and neurobiological factors. Publication 1 addressed the question of whether mothers can identify their children by body odor, whether they prefer this odor and how it interacts with genetic, hormonal factors and the child's developmental status. For this purpose, N = 164 mothers with their biological children (N = 226 children between 0 and 18 years) were included in the study and evaluated the body odors of their own and unfamiliar children, which differed in their developmental stage and genetic similarity. Genetic similarity was mapped via the human leukocyte antigen (HLA) profile, the developmental status was determined on the basis of the steroid hormone concentration (testosterone, estradiol) and a standardized assessment of the pubertal status. The results showed that the mothers were able to identify their own child's odor above chance level and preferred this odor. This was true for all age groups with the exception of early puberty. At this age (9-13 years), mothers could neither identify the odor of their child nor preferred it to unfamiliar body odors. For the body odor ratings of their own sons, the decrease in preference was associated with an increase in testosterone level. In older children (14-18 years), maternal ratings resembled those before puberty suggesting that the mothers get used to the altered body odor of their child during this period and thus, the familiarity of the odor plays an important role for perception. In addition, the results demonstrated that genetic similarity is transported via body odors: Although the preference for the odor of one's own child was globally observed, pairwise comparisons showed that the ratings for the own child ́s odor did not differ significantly from the evaluation of a same-aged and HLA-similar child. This suggests that genetic similarity has a positive effect on odor assessment in the context of parent-child bonding. In the second publication, in the same sample it was examined whether mothers are able to classify the child's developmental status on the basis of body odors and which predictors are decisive for the classification. Therefore, the mothers were asked to assess the age group of the child who was the odor donor. The results revealed that mothers are able to detect the developmental status (pre- vs. postpubertal) with an accuracy of 64% and tend to classify body odors as prepubertal. The maternal classification performance was better when they rated odor samples from the same age group as their own child. The perceptual evaluation of the samples (pleasantness, intensity) as well as the assessed pubertal status predicted the development-related classification of an odor, while the child ́s steroid hormone concentration had no effect on it. The third publication of this doctoral thesis served as a methodical pilot study for the specific examination of the influence of baby odors on neural processing in the maternal brain. From other modalities, it is known that infantile stimuli transport cuteness leading to reward-related neural correlates. However, this has scarcely been investigated for baby odors so far. Body odor presentation in functional magnetic resonance imaging (fMRI) is challenging due to stimulus properties and methodological difficulties. To date, only a few studies exist on the neural processing of body odors without a uniform convention on a suitable stimulus presentation. The aim of this thesis was to develop an efficient design that optimally maps neural activity in response to baby body odors. For that reason, two stimulus presentations were compared which differed in presentation mode, duration and frequency. The short, continuous stimulus presentation revealed stronger global activations compared to a long, pulsed presentation, thus it is recommended as a design to obtain robust neuronal correlates. However, differential effects were observed depending on the brain regions, which is why the design should be specifically adapted to the regions of interest, and length, duration and type of stimulus presentation should be considered carefully. The short presentation design was used in the follow-up fMRI study. This illustrated that baby body odors activate reward areas and a network encoding cuteness and motivation to care (pleasure network). The recruitment of this network predicted how pleasantly mothers rated their own baby's odor. In contrast to the behavioral data from publication 1, which showed a clear preference for one's own child, the neural responses did not differ between one's own or an unfamiliar baby ́s odor. Therefore, the universality of baby odor as a stimulus conveying cuteness must be systematically examined in subsequent studies. In summary, this doctoral thesis reveals that children ́s body odors function as chemosignals in the mother-child relationship and mediate both the identification of the own child and affective components. In addition, it was observed that information about genetic similarity and the child's developmental status are transcribed in body odors. It remains to be explored which factors at the molecular level actually determine changes in body odor. Future investigations using chemosensory profile analyses may clarify this question. Beyond that, longitudinal studies are necessary in order to depict the associations presented here over the course of individual development and thus enabling the derivation of mechanisms of the olfactory mediated parent-child relationship. In the long term, this information should help to generate strategies for promoting the parent-child relationship and to extend existing interventions (such as neurofeedback training) to olfactory stimuli.:Inhaltsverzeichnis Danksagung 4 1 Zusammenfassung 6 2 Summary 9 3 Einführung in die Thematik 12 4 Studienziele: Abgeleitete Forschungsfragen und Hypothesen 21 5 Methodik der Untersuchungen 23 6 Zusammenfassung der Ergebnisse 26 7 Diskussion und Ausblick 29 8 Literaturverzeichnis 40 Anhang I. Verzeichnis der wissenschaftlichen Veröffentlichungen, Konferenzbeiträge und andere Leistungen A Veröffentlichungen der Dissertation und dazugehörige Angaben B Weitere Veröffentlichungen während der Promotionsphase C Konferenzbeiträge und andere Leistungen während der Promotionsphase II. Letters of Acceptance III. Erklärung zur Eröffnung des Promotionsverfahrens IV. Bestätigung der Einhaltung der folgenden aktuellen gesetzlichen Vorgaben

[ES] La presente tesis doctoral titulada "Nanodispositivos mesoporosos híbridos funcionalizados con enzimas para detección, liberación controlada y comunicación molecular" se centra en el diseño, preparación, caracterización y evaluación de distintos nanodispositivos híbridos orgánico-inorgánicos utilizando como soporte nanopartículas tipo Janus de oro y sílice mesoporosa, que se equipan con enzimas, especies fluorescentes y puertas moleculares. Como conclusión general, los estudios realizados muestran que la incorporación de enzimas sobre nanopartículas permite introducir funciones de reconocimiento con alta especificidad y diseñar nanodispositivos avanzados para distintas finalidades. La combinación de nanopartículas híbridas con grupos orgánicos como puertas moleculares, efectores enzimáticos y especies cromo- fluorogénicas o fármacos puede resultar muy versátil; y se espera que los resultados obtenidos puedan inspirar el desarrollo de nuevos materiales inteligentes con aplicación en distintas áreas como la nanomedicina y la detección de moléculas de interés. / [CAT] La present tesi doctoral titulada "Nanodispositius mesoporosos híbrids funcionalitzats amb enzims per a detecció, alliberació controlada i comunicació molecular" es centra en el disseny, preparació, caracterització i avaluació de distints nanodispositius híbrids orgànic-inorgànics utilitzant com a suport nanopartícules tipus Janus d'or i sílice mesoporosa, que s'equipen amb enzims, espècies fluorescents i portes moleculars. Com a conclusió general, els estudis realitzats mostren que la incorporació d'enzims sobre nanopartícules permeten introduir funcions de reconeixement amb alta especificitat i dissenyar nanodispositius avançats per a distintes finalitats. La combinació de nanopartícules híbrides amb grups orgànics com portes moleculars, efectors enzimàtics i espècies cromo-fluorogèniques o fàrmacs pot resultar molt versàtil; i s'espera que els resultats obsessos inspiren el desenvolupament de nous materials intel·ligents amb aplicació en distintes àrees com la nanomedicina i la detecció de molècules d'interés. / [EN] This PhD thesis entitled "Enzyme-functionalized hybrid mesoporous nanodevices for sensing, controlled release and molecular communication" is focused on the design, synthesis, characterization and evaluation of several hybrid organic-inorganic nanodevices using Janus gold-mesoporous silica nanoparticles as scaffolds, equipped with enzymes, fluorescent species and molecular gates. In conclusion, these studies show that the incorporation of enzymes on nanoparticles allows to introduce recognition capabilities with high specificity and to design advanced nanodevices for different purposes. The combination of hybrid nanoparticles with organic groups such as molecular gates, enzymatic effectors and chromo-fluorogenic species or drugs can be very versatile; and we hope that the obtained results inspire the development of new smart materials with application in different areas such as nanomedice and sensing. / Llopis Lorente, A. (2019). Enzyme-functionalized hybrid mesoporous nanodevices for sensing, controlled release and molecular communication [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/117612 / TESIS

The thesis consists of two parts. The first part introduces the topic of human chemical communication and reviews current evidence on individual variation in human body odour and its perception. This part is framed by sexual selection theory. In the first chapter, the concept of the theory of communication is introduced followed by a discussion on the specifics of chemical communication. Next, the formation of individually specific body odour signatures with reference to skin glands, their volatile products and the subsequent metabolization by skin microflora is described. The next chapters are dedicated to selected interindividual body odour cues such as sex and kin recognition, genetic compatibility in genes of Major Histocompatibility Complex, and health and reproductive status in a mate choice context. Furthermore, interactions between perfumes and body odours are discussed. Finally, methods of body odour sampling are introduced and a rationale behind presenting individual samples or body odour blends is discussed. The second part is comprised of six scientific papers, specifically three reviews and three empirical studies. Review papers summarize factors affecting human body odour quality with emphasis on diet and affective states. The first text shows that human body odours contain cues to...

The traditional assumption that perfumes are used only for masking of the body odour was doubted by the results of recent researches. In spite of the fact that the biological role of human body odour is very important, it is likely that perfumes are not in conflict with the body odour but there is mutual cooperation. Individuals could choose the scent which goes with their body odour instead of hiding it. In the theoretical part of this thesis I describe a dual inheritance theory, which is a basic concept for our research. We have been concerned with the interaction between biological effects of body odour and social effects of perfumes. Moreover, we have worked on the assumption that the body odour of relatives is similar. We have tested if people are better in their choice of the appropriate perfume for their relatives than for anyone else, in this case partners. Which perfume fits into which body odour was assessed by the independent evaluators of opposite sex because one of the main objectives of chemical signalling is to attract a potential partner. Surprisingly, the results of our research have shown that in the case of couples, the samples of the body odour and perfumes chosen by female partners were slightly better evaluated than the perfumes chosen by men themselves. Moreover, there were...

Changement climatique, urbanisation, invasions biologiques et hybridation interspécifique constituent des menaces majeures pour la biodiversité. L'objectif de cette thèse a été d'évaluer leurs effets interactifs sur les fourmis du genre Tetramorium de la vallée du Rhône, France. Près de 2000 colonies de 5 espèces de Tetramorium ont été étudiées. Les résultats démontrent que le climat et l'urbanisation limitent leurs aires de répartition et que l'urbanisation impacte leurs distributions simultanément à plusieurs échelles spatiales. Les probabilités d'occurrence de T. immigrans dépendent de l'interaction entre facteurs climatiques et urbains. Quatre populations génétiques de T. immigrans résultent de plusieurs introductions de sources externes, ce qui signifie que cette espèce n’est pas indigène dans les zones urbaines du nord. T. immigrans et T. caespitum produisent des hybrides dans les zones de sympatrie. La détection de rétrocroisements et de discordances entre ADN mitochondrial et nucléaire suggère que cette hybridation conduit à une descendance fertile, donc de l’introgression. Les profils d'hydrocarbures cuticulaires sont bien différenciés entre les deux espèces et sont corrélés à une agression forte contre les hétérospécifiques. Ces espèces présentent un système d'accouplement monogyne polyandre, dans lequel les reines hybrides mais pas les pères hybrides contribuent à la descendance hybride. Les études sur l'hybridation interspécifique n'ont qu'occasionnellement concerné les changements globaux ou invasions biologiques. Ces concepts eux-mêmes sont peu étudiés ensemble et leurs effets interactifs sont négligés dans la littérature actuelle. La prise en compte de ces interrelations et l'exploration de plusieurs échelles sont essentielles pour mieux comprendre les processus qui génèrent des modèles d'échange génétique / Climate change, urbanization, biological invasions and interspecific hybridization are major current threats to biodiversity. The objective of this thesis was to assess their interactive effects in Tetramorium ant species from the Rhône-Saône valley in France. Using a broad array of tools from several fields of biology, I worked on almost two thousand colonies from five Tetramorium species including T. immigrans and T. caespitum. Both climate and urbanization limited the ranges of these species, and urbanization impacted their distributions simultaneously at several spatial scales. The occurrence probabilities of T. immigrans depended on the interaction between climatic and urban factors. Several introductions from external sources seemed to have given rise to four genetically distinct populations of T. immigrans, making it likely that it is not native to the northernmost urban areas. Hybridization and introgression between T. immigrans and T. caespitum was frequent in zones of sympatry, and the detection of nuclear DNA backcrosses and mitochondrial-nuclear DNA discordance between the two species suggested that hybridization lead to fertile offspring (i.e. introgression). Cuticular hydrocarbon profiles were clearly differentiated between the two species and were correlated to heightened interspecific aggression against heterospecifics. Both species presented a monogyne polyandrous mating system, in which hybrid queens but no hybrid fathers contributed to hybrid offspring. Few studies on interspecific hybridization have addressed global change or biological invasions. These concepts are infrequently studied together, and their interactive effects are overlooked in the current literature. Taking these interrelationships into account and exploring multiple scales are essential to better understand the processes that generate patterns of genetic exchange

Made available in DSpace on 2015-04-22T22:12:47Z (GMT). No. of bitstreams: 1 Erika Portela de Lima Silva.pdf: 770664 bytes, checksum: b6b83b117ec0ee88c1214d68278a1c6f (MD5) Previous issue date: 2012-03-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The study of the interaction between pairs of species is the basis for understanding the coexistence in biological communities. The ability to select habitats may be important in the coexistence of competing species and in the interactions between predators and prey. This selection is often associated with the capacity of perception of chemical cues of prey and structural components of the habitat. The perception and response to chemical cues have been demonstrated for wandering spiders, but only for the family Lycosidae in temperate regions. The soil is among the structural factors that affect habitat selections in spiders, but this has been demonstrated only for burrowing spiders. In tropical forests, high biological diversity and the rapid degradation may hinder the recognition of cues from multiple species of predators and prey. Habitat selection based on different soil types may be important in this ecosystem where drastic soil alternations are common between nearby sites, especially if the soil type is related to some important resource. We assessed the response of two species of wandering spiders in a forest in central Amazonia, Ctenus amphora Melo-Leitão 1930 and Ctenus crulsi Melo-Leitão 1930 (Araneae, Ctenidae) (1) to chemical cues from potential prey, crickets Gryllus assimilis (Fabricius, 1775) (Orthoptera, Gryllidae) and termites Syntermes Holmgren, 1910 (Isoptera, Termitidae), (2) to chemical cues from predators, spiders Phoneutria Perty, 1833 (Araneae, Ctenidae) and army ants Labidus praedator (Fr. Smith, 1858 ) (Formicidae, Ecitonini), (3) and two soil types, sandy and clay soils on which the distribution and abundance of these species diverge. For the first two evaluations we compared the proportion of time on filter paper with and without the chemical cues, to the third evaluation we compared the proportion of time on the two types of soil. These observations were performed over 09 hours with 54 observations per spider at intervals of 10 minutes. A total of 65 spiders were observed in the experiment with prey (termites cues, crickets cues and control), 36 in the experiment with predators Phoneutria (Phoneutria cues and control), 15 in the experiment with army ants (army ants cues and control) and 67 spiders in the soil experiment. The two species spent significantly longer time on the filter paper with chemical cues from the 9 prey, which indicates that the perception of cues can be important for the foraging of the two species. Although C. crulsi consumes far more Syntermes termites in natural conditions, the two species did not differ in the proportion of time on the cues from termites in comparison to the time on cues from crickets, sugesting that they do not differ in the ability to detect the chemical from these prey. No antipredator response was detected for the two species of Ctenus. Therefore, there is no evidence that the coexistence of these species is facilitated by differences in the perception of chemical cues. Only C. crulsi presented preference to a type of soil, the clay soil, which coincides with the type of soil where it is found in abundance in nature. This response of C. crulsi can influence the distribution of C. amphora, due to interactions of these species. Therefore, the perception of soil type is probably an important proximal factor in determining the distribution patterns and dynamics of coexistence of these species. / O estudo da interação entre pares de espécies é uma das bases para a compreensão da coexistência em comunidades biológicas. A capacidade de selecionar habitats pode ser importante tanto na coexistência de espécies que competem entre si como nas interações entre predadores e presas. Esta seleção está muitas vezes associada à capacidade de percepção de pistas químicas de presas e de componentes estruturais do habitat. A percepção e resposta a pistas químicas já foram demonstradas para aranhas errantes, mas somente para a família Lycosidae em regiões temperadas. O solo está entre os fatores estruturais que influem na seleção de habitat em aranhas, mas isto apenas foi demonstrado para aranhas construtoras de toca. Em florestas tropicais, a alta diversidade biológica e a rápida degradação podem dificultar o reconhecimento de pistas das múltiplas espécies de predadores e de presas. A seleção de habitat com base nos diferentes tipos de solo pode ser importante neste ecossistema em que são comuns alternâncias drásticas de solos entre locais próximos, especialmente se o tipo de solo estiver relacionado com algum recurso importante. Avaliamos a resposta de duas espécies de aranhas errantes de uma floresta na Amazônia central, Ctenus amphora Melo-Leitão 1930 e Ctenus crulsi Melo-Leitão 1930 (Araneae, Ctenidae) (1) às pistas químicas de potenciais presas, grilos Gryllus assimilis (Fabricius, 1775) (Orthoptera, Gryllidae) e cupins Syntermes Holmgren, 1910 (Isoptera, Termitidae); (2) às pistas químicas de predadores, aranhas Phoneutria Perty, 1833 (Araneae, Ctenidae) e formigas de correição Labidus praedator (Fr. Smith, 1858) (Formicidae, Ecitonini); (3) e dois tipos de solo, solo arenoso e argiloso, sobre os quais a distribuição e abundância destas espécies diverge. Para as duas primeiras avaliações, comparamos a proporção de tempo sobre recortes de papel filtro com e sem as pistas químicas, para a terceira avaliação comparamos a proporção de tempo sobre os dois tipos de solo. Estas observações foram realizadas ao longo de 09 horas com 54 observações por aranha realizadas em intervalos de 10 minutos. Foram observadas 65 aranhas no experimento com as presas (pistas de cupins, grilos e controle), 36 aranhas no experimento com Phoneutria (pistas de Phoneutria e controle), 15 no experimento com formigas de correição (pistas de 7 formiga e controle), e 67 aranhas no experimento de solos. As duas espécies permaneceram significativamente mais tempo sobre as pistas de presas, indicando que a percepção de pistas pode ser importante para o forrageio das duas espécies. Apesar de C. crulsi consumir muito mais cupins Syntermes em condições naturais, as duas espécies não diferiram na proporção do tempo sobre as pistas de cupins em comparação ao tempo sobre pistas de grilos, indicando que não diferem na capacidade de detectar quimicamente estas presas. Não foi detectada resposta antipredatória pelas duas espécies de Ctenus. Portanto, não encontramos evidências que a coexistência destas duas espécies seja facilitada por diferenças na percepção de pistas químicas. Apenas para C. crulsi foi encontrada resposta de preferência a um dos tipos de solo, o argiloso, que coincide com o tipo de solo onde ela é encontrada em maior abundância na natureza. A resposta de C. crulsi pode influir na distribuição de C. amphora, em função das interações destas duas espécies. Portanto, a percepção do tipo de solo provavelmente é um fator proximal importante a determinar os padrões de distribuição e a dinâmica da coexistência destas espécies.

Il est plus que nécessaire aujourd'hui d'apporter de nouvelles avancées dans le domaine des traitements anti-cancéreux. Il est également bien établi que dans la famille des produits bioactifs d'origine naturelle, les composés azotés sont largement représentés et ce dans la plupart des classes thérapeutiques. Aussi, ces travaux se sont concentrés sur l'obtention de composés azotés bioactifs d'origine naturelle en ciblant des produits cytotoxiques. Ce travail est divisé en trois parties. Tout d'abord, nous avons choisi d'étudier les souches bactériennes associées aux lichens. En effet, le réseau symbiotique complexe que représente le lichen est d'un grand intérêt pour l'identification de nouvelles souches bactériennes avec des potentiels de production de composés d'intérêt et notamment d'un point de vue biotechnologique. Dans un premier temps, un travail d'optimisation des conditions de culture d'une souche d'Actinobactérie, Nocardia sp, isolée à partir d'un lichen, Collema auriforme, a été réalisé dans le but de favoriser la production de composés cytotoxiques. Dans un second temps, l'étude bactériochimique de cette souche a permis l'isolement de 13 composés azotés, dont deux dicétopipérazines bromées qui sont nouvellement décrites ainsi que des dérivés puriques et pyrimidiques. Enfin, un travail de ciblage des composés actifs, par réseaux moléculaires et analyses multivariées de données de spectrométrie de masse, a mis en évidence des molécules potentiellement responsables de l'activité. La seconde partie de ce travail apporte des éléments de réponse sur la nature de la relation bactérie-lichen en observant les effets de certains composés lichéniques sur la croissance et le métabolisme bactériens ainsi qu'en étudiant le potentiel de biotransformation de trois bactéries associées aux lichens (une Firmicutes, une beta-protéobactérie et une Actinobactérie). Les souches sélectionnées ont toutes montré des capacités de biotransformation des composés lichéniques testés (notamment acide usnique et méthyl-beta-orcinolcarboxylate). Enfin, la troisième partie de ce travail décrit une nouvelle méthode d'obtention de composés azotés bioactifs d'origine naturelle par la voie synthétique. Ainsi, une nouvelle méthode de synthèse de la bgugaine, la coniine et la tylophorine a été étudiée et a permis d'obtenir la N-Boc-2-heptylpyrrolidine, démontrant la possibilité de synthétiser des alcaloïdes de type pyrrolidine et d'envisager la synthèse des cibles sélectionnées. / New advances in anti-cancer therapeutic are today more than necessary. It is also well establish that amongst bioactive natural compounds, nitrogen compounds are widely represented and that, in all therapeutic classes. Thus, these works are focused on obtaining bioactive nitrogen compounds from natural sources targeting cytotoxic products. This work is divided in three parts. First of all, we chose to study bacterial strains associated with lichen. Indeed, lichens are a complex symbiotic network of great interest for the discovery of new bacterial strains. Especially for strains with a great biotechnological potential particularly in the production of compounds of interest. At first, culture conditions of an Actinobacteria, Nocardia sp, isolated from a lichen, Collema auriforme, were optimised in order to produce cytotoxic compounds. Secondly, the study of the secondary metabolites patterns of this strain allows the isolation of 13 nitrogen compounds, including 2 brominated diketopiperazines which are newly described, along with purine and pyrimidine derivatives. Finally, a targeting work of bioactive compounds was realised by establishing molecular networking and multivariate analysis of mass spectrometry data, highlighting molecules potentially responsible for activity. The second part of this work bring some answers about the nature of the relationship between bacteria and lichen, by observing effect of some lichen compounds on bacterial growth and metabolism as well as studying the biotransformation potential of three lichen-associated bacteria (a Firmicutes, an beta-proteobacteria and an Actinobacteria). All selected strains showed ability for the biotransformation of tested lichen compounds (usnic acid and methyl-beta-orcinolcarboxylate). Finally, the third part of this work describes a new method for obtaining bioactive nitrogen compounds by a synthetic way. Thus, a novel synthesis method of bgugaine, coniine and tylophorine was studied and led to N-Boc-2-heptylpyrrolidine, showing the possibility to synthesize pyrrolidine alkaloids and to consider the synthesis of selected targets.

La colonie d’abeille (Apis mellifera L.) est une société complexe où les individus interagissent entre eux, notamment par le biais de phéromones. L’étude de cette communication chimique est indispensable à la compréhension des régulations sociales mises en place dans la colonie. Chez l’abeille, plus de 50 substances chimiques avec des effets incitateurs ou modificateurs sur la colonie ont été identifiées. Malgré ces découvertes, de nombreux travaux sont à accomplir pour mieux comprendre ce système de communication particulier.La problématique de cette thèse vise à caractériser l’histoire de vie d’une phéromone majeure l’Oléate d’Ethyle (EO), qui permet d’optimiser l’équilibre nourrices / butineuses dans la colonie. Parallèlement, d’autres recherches ont été entreprises, notamment l’étude de la communication chimique de la reine et du couvain, chez qui seulement deux phéromones ont été identifiées avec des effets pléiotropiques dans la colonie.Nos résultats ont mis en évidence une production variable d’EO par les ouvrières, en fonction de l’environnement de la colonie. La production de cette molécule chimique dans la colonie peut également être modifiée par un stress : des abeilles parasitées par du Nosema spp. ont une production anormalement élevée d’EO. En outre, cette molécule phéromonale est transmise des butineuses vers les nourrices par contact cuticulaire et par le pollen.Pour la compréhension de la communication entre la reine et les ouvrières, nos résultats montrent que la reine utilise d’autres composés phéromonaux puissant en redondance de la QMP pour orienter la construction de cire, le phénomène de cour et l’inhibition des ovaires des ouvrières.Chez le couvain, nous avons identifié un composé phéromonal volatil, le E-ß-ocimène, produit majoritairement par les jeunes larves, inhibant le développement des ovaires des ouvrières et accélérant leur maturation comportementale.Ces études nous ont permis d’avoir une connaissance plus précise de la communication chimique au sein de la colonie. Ainsi nous expliquons par deux théories le rôle de la complexité et de la redondance phéromonale de la colonie d’abeilles / In the honeybee colony (Apis mellifera L.) studies of the chemical communication are essential to understand social regulations. In the honey bee colony more than 50 chemical substances with releaser and primer effects have been identified. Despite years of research on this type of communication, significant work remains to be done.In this thesis, the aim is to characterize the dynamics of a major pheromone: ethyl oleate (EO), which optimizes the balance between nurses and foragers in the colony. In addition, we initiated research on the queen and brood chemical communication in which only two pheromones have been identified in the colony.We have demonstrated that EO production by workers varies under different colony environment. EO production can also be modified by stress; honey bees parasitized by the Nosema spp. have abnormally high EO production. In addition, we identified that EO is transmitted from foragers to nurses by contact (cuticle and pollen).For the queen, our results indicate that the queen uses multiple redundant pheromones (QMP and other unknown compounds), that affect wax construction, retinue behaviour and worker ovary inhibition.For the brood we have identified a volatile pheromone E-ß-ocimene produced mostly by the young larvae to inhibit the development of workers ovaries and accelerate workers’behavioural maturation.With these studies we clarify some aspects of what is known about chemical communication in the honey bee colony. Then we try to explain the role of complexity and redundancy of pheromones in the honey bee colony by two theories