Calcified teeth comprise the hardest vertebrate tissues (enamel and dentine) and they undoubtedly represent a key evolutionary advantage for their users. Nevertheless, teeth have been lost many times and they were often replaced by keratinised structures, such as rhamphoteca of birds or of turtle beaks, or labial tooth structures and scratching laminae of many aquatic vertebrates. This thesis is intended to analyse levels of similarities among keratinised teeth and structures of three phylogenetically distinct lineages of amniotes and to compare them to morphogenesis of true teeth. Detailed developmental and structural characteristics of oral apparatus possessing keratinous teeth of lampreys, Loricarid cathfishes and frog tadpoles were identified and distinguished. Keratinous structures of cathfishes include only horny projections from single cell surface (unculi), whereas in frog tadpoles both horny jaw sheet and unicellular labial teeth composed by cell columns were found; the lamprey oral apparatus consists of multicellular teeth and laminae instead. Disparate phenotypes of these keratinous structures can, however, developmentally derive from a general process of keratinization: morphological disparities arise by distinct gradual accretion of keratinous layers and collumns during development. My...

The work presented herein is a method optimization for biomolecule detection and identification using Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry Imaging (MALDI-MSI). MALDI-MSI is a unique form of mass spectrometry that is highly multiplexed; it can simultaneously retain location information of the mass of multiple ions, allowing for correlation of morphology or pathology to reconstructed ion heat maps. There were three main objectives for the research - 1) A method optimization of sample preparation techniques for bottom-up proteomic MALDI-MSI was performed. This included the optimization of tissue wash steps, trypsin digestion incubation times, and matrix deposition techniques. The results included identifying the appropriate pH for the wash steps to optimize trypsin digestion, an overnight trypsin incubation to allow for complete digestion, and the inclusion of MCAEF – Matrix Coating Assisted by an Electric Field – during matrix coating for enhanced spectra. 2) An unbiased statistical data processing workflow for simultaneous processing of multiple datasets was performed. This was done using a thyroid hormone treated tadpole dataset to gain insight into the metabolism of anuran metamorphosis. Results found included a finalized data processing workflow that detected 5000 metabolite features from five organs were detected in pre-metamorphic tadpoles. Of these detected metabolites, 136 were significantly affected upon exposure to thyroid hormone and 64 metabolites were putatively identified. 3) A sample preparation technique for metabolomic analysis of formalin-fixed paraffin embedded (FFPE) colorectal liver metastasis samples was performed. Results included the importance of using a high mass resolution mass spectrometer while emphasizing more appropriate use of fresh-frozen tissue sections for metabolomic analysis. / Graduate

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Cannibalism is a seemingly aberrant interaction, appearing counter to the fitness of individuals. Yet cannibalism is not overly uncommon, and naturally occurs among aquatic organisms, including larval amphibians. In temporary wetlands larval amphibians are in a race to complete metamorphosis before their aquatic habitat disappears. When intraspecific competition intensifies, eating conspecifics may represent a beneficial if not necessary strategy. The research presented within this thesis aims to characterize factors that influence cannibalism within populations of larval amphibians. Wood frog tadpoles (Lithobates sylvaticus) were used to test potential benefits of cannibalism as a diet, determine if dietary quality and nutritional stress influence cannibalism, and investigate the roles of competition and chemical cues in influencing cannibalism. Larval long-toed salamanders (Ambystoma macrodactylum), and ringed salamanders (A. annulatum) were used to investigate a functional link between trophic polymorphism and cannibalism in natural populations. Results suggest that perceived increases in competition may stimulate some individuals to become less risk averse, and more aggressive, which may in turn facilitate cannibalistic behaviour. Cannibalism itself provided only conditional benefits to larval wood frogs, rather than the optimal growth that would be expected from an ideal diet. However, this may have been the result of individual variation in response to the diet and/or conspecific cues as opposed to a nutritional deficit. In conditions where tadpoles could perceive increased competition they altered their behaviour and morphology in ways that may improve their foraging success and potentially promote cannibalism. Finally, a functional link appears to exist between head morphology and cannibalism in natural wetlands. However, the appearance of this morphology appears related to conditions that may facilitate increased population densities through rapid pond drying.

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Un des avantages des lasers femtosecondes de haute puissance (TW-PW) est de pouvoir obtenir, au foyer d'une optique focalisante, des intensités très élevées atteignant jusqu'à 10^22W.cm^-2 soit un champ électrique de 2.7 PV.m^-1. Pour cela, ces chaînes lasers délivrent nécessairement des faisceaux de grands diamètres (jusqu'à 40 cm) et des impulsions très courtes (de l'ordre de la dizaine de femtosecondes). En conséquence, les propriétés spatiales et temporelles de l'impulsion ne sont généralement pas indépendantes. Ce type de dépendance, appelée couplage spatio-temporel, a pour conséquence d'augmenter la durée d'impulsion et la taille de la tache focale, ce qui peut conduire à une diminution notable de l'intensité maximale au foyer. Les dispositifs de métrologie couramment utilisés sur ces chaînes lasers femtosecondes de haute puissance ne permettent de mesurer les profils spatial et temporel de l'impulsion que de façon indépendante.L'objectif de cette thèse était de développer des techniques permettant de mesurer les couplages spatio-temporels afin de pouvoir quantifier leur effet et de les corriger dans l'optique d'obtenir l'intensité maximale au foyer. Ainsi, nous avons tout d'abord adapté une technique de caractérisation spatio-temporelle existante à la mesure de lasers TW. Afin d'éviter les contraintes induites au foyer, comme celles liées aux fluctuations de pointé, les mesures ont été réalisées sur le faisceau collimaté. Ajouter une source de référence en parallèle du dispositif initial, nous a aussi permis de prendre en compte les artéfacts de mesure dus aux variations thermiques et mécaniques affectant l'interféromètre. Grâce à cette amélioration, il est possible de reconstruire le profil spatio-temporel complet du faisceau, en particulier son front d'onde.Cependant, les limitations induites par cette technique, nous ont conduit à développer un nouveau dispositif de mesure. Basé sur une corrélation croisée, cette technique consiste à faire interférer le faisceau laser à caractériser avec une partie de ce dernier, suffisamment petite pour ne pas être distordue spatio-temporellement. Nous avons également mis en œuvre une variante de ce dispositif permettant une mesure mono-coup selon une dimension transverse de l'impulsion.A l'aide de ces différentes techniques, nous avons pu caractériser, pour la première fois, plusieurs chaînes lasers TW. Les mesures réalisées ont mis en lumière l'existence de couplages spatio-temporels résiduels conduisant à une baisse significative de l'intensité pic au foyer. Ces résultats montrent qu'il est indispensable de caractériser spatio-temporellement des chaînes lasers femtosecondes de haute puissance dans l'optique d'obtenir l'intensité maximale au foyer. / One of the advantages of high-power femtosecond lasers (TW-PW) is to obtain, at the focus of a focusing optic, very high intensities up to 10^22W.cm^-2 (i.e. an electric field of 2.7 PV.m^-1. Therefore, these lasers chains necessarily deliver beams with large diameter (up to 40 cm) and very short pulses (of the order of tens of femtoseconds). As a consequence, the spatial and temporal properties of the pulse are generally not independent. Such dependence, called spatial-temporal coupling has the effect of increasing the pulse duration and the size of the focal spot, which can lead to a significant reduction of the maximum intensity at the focus. Metrology devices commonly used on these high-power femtosecond lasers allow retriving the spatial and temporal profiles of the pulse only in an independent manner. The aim of this thesis was to develop techniques for measuring spatiotemporal couplings in order to quantify their effect and correct them in order to obtain the maximum intensity at focus. First of all, we adapted an existing technique of spatio-temporal characterization to the measurement of TW lasers. To avoid the issues induced at the focus, such as those related to jittering, measurements were performed on the collimated beam. By adding a reference source to the original device, we managed to take into account the measurement artifacts due to thermal and mechanical variations affecting the interferometer. With this improvement, it was possible to reconstruct the complete spatio-temporal profile of the beam, particularly its wavefront. However, the limitations imposed by this technique led to the development of a new measurement device. Based on a cross-correlation, this technique consists of making the laser beam to interfere with a part of itself, small enough not to be spatiotemporally distorted. We have also implemented a variant of this device for a single-shot measurement along one transverse dimension of the pulse. Using these techniques, we performed the very first characterization of several TW laser chains. The measurements have revealed the existence of residual space-time couplings leading to a significant decrease of the peak intensity at focus. These results show that it is essential to spatio-temporally characterize high power femtosecond laser chains to obtain the maximum intensity at focus.

Amphibien sind global von einem Artenrückgang betroffen, wobei Ausbeutung bestimmter Froscharten einer der Gründe hierfür ist. Die vorliegende Arbeit gibt erstmals Einblick in Form, Umfang und Auswirkungen der extensiven Nutzung von Fröschen in Westafrika. Interviews mit Konsumenten und in den Froschhandel Involvierten ermöglichten die Nutzung und den Handel von Fröschen in Burkina Faso, Benin und Nigeria zu quantifizieren und zu bewerten. Während der Froschhandel in Burkina Faso eher auf lokaler Ebene abläuft, ließ sich im Norden Benins und in Nigeria ein intensiver grenzüberschreitender Handel nachweisen. Um einen möglichen anthropogenen Einfluss auf natürliche Amphibien Gemeinschaften zu ermitteln wurden natürliche Gewässer untersucht. Auf Kaulquappenebene wurden Artenverluste und die sich daraus ergebenen Konsequenzen studiert. Temporäre Savannengewässer wurden in zwei Untersuchungsgebieten in Burkina Faso untersucht, wobei die Gewässer jeweils in Gebieten mit unterschiedlichem Störungsgrad lagen: in Dörfern mit Froschfang, gegenüber in geschützten Gebieten mit Froschfangverbot. Generell konnte in den anthropogen gestörten Gebieten ein niedrigerer Artenreichtum mit gleichzeitig veränderter Zusammensetzung der Artengemeinschaften konstatiert werden. Für diese Unterschiede waren primär anthropogen veränderte Habitatfaktoren verantwortlich. Aber auch das Froschfangen kann Grund für veränderte Kaulquappengemeinschaften sein. Um Voraussagen für mögliche resultierende Konsequenzen treffen zu können ist es wichtig die ökologische Rolle betroffener Arten im System zu kennen. In künstlichen Gewässern wurde die trophische Position von vier Kaulquappenarten mit jeweils unterschiedlicher Nahrungsstrategie und die Konsequenzen ihres Verlustes für die restlichen Kaulquappen und bestimmter Ökosystemfaktoren, wie Moskitolarven, ermittelt. Die vier Kaulquappenarten unterschieden sich in ihrer trophischen Position. Zudem konnten komplexe Interaktionen zwischen den vier Fokusarten aufgedeckt werden, wobei die trophische Position je nach Gemeinschaft wechseln konnte. So konnte ich mit meiner Arbeit zeigen, dass der Verlust nur einer Art essentielle Konsequenzen für Amphibien Gemeinschaften und ökosystemische Prozesse haben kann. Chapter 1 Summary II I / Amphibian populations are declining world wide, with overexploitation being one of the main reasons. In Africa frog exploitation has not been in the focus of scientific research yet. Herein I report first investigations of the frogs use from West Africa. Interviews with consumers and participants in the frog trade facilitated to quantify and evaluate the use and trade of frogs in Burkina Faso, Benin and Nigeria. In Burkina Faso the frog trade mainly took place on a local scale, whereas in northern Benin and Nigeria, I detected an intensive cross-border trade. In an empiric approach I examined if a possible anthropogenic impact could be found in natural tadpole communities. In two study regions in Burkina Faso I compared natural freshwater ponds situated of different disturbance regimes: ponds in and around villages, where frogs were collected for consumption, and in protected areas, where frog harvest was prohibited. Species richness was lower and species composition differed in anthropogenic disturbed areas. Anthropogenic altered habitat factors were mainly responsible for these differences. However, harvesting of adult amphibians could have affected tadpole composition and occurrences of particular species as well. To evaluate the effect of the potential loss of a species a better understanding of its ecological role is important. Therefore, I conducted an experiment to investigate the trophic position of four tadpole species and the consequences of species exclusion for the remaining species and on certain ecosystem parameters such as mosquito larvae. Isotopic analyses revealed that tadpole species differed in their trophic position. Various complex interactions between species occurred including shifts in trophic positions due to tadpole species composition. Thus the loss of just one species can have crucial consequences for the remaining amphibian community and furthermore for ecosystem processes. Chapter 1 Summary II I

Triops cancriformis (Crustacea: Notostraca) occurs in ephemeral habitats like rain pools or floodplain pools distributed over a large geographical range. The named habitats are disturbed by human impacts and, consequently, T. cancriformis is endangered throughout its distribution range. In the present thesis the populated habitats and threats are characterised and further morphological and genetic variations detected among and within European populations are reported. On the basis of recent investigations it is shown that T. cancriformis subspecies separation is hampered by an individual variability which points to the necessity of species revision. The analysis of mitochondrial gene sequence data suggests that the species has colonised most of Europe very recently. The advantage of a complex reproductive strategy in T. cancriformis in this process is discussed. The population structure resolved with nuclear DNA markers highlights that there is low allelic diversity among and within populations compared to other Branchiopoda (Daphnia). By means of the present study it can be shown that habitat conservation is most important to protect T. cancriformis.

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