Die Geruchswahrnehmung von Insekten ist an die spezifischen Anforderungen der Ökosysteme angepasst, in denen sie leben. Von Insekten wahrgenommene Volatile können Informationen über den Ort eines geeigneten Ovipositionsplatzes oder den physiologischen Zustand einer Wirtspflanze geben. Da der Geruchssinn der Insekten sich seit Millionen von Jahren an die Wahrnehmung bestimmter Volatile angepasst hat, können die wahrgenommenen Volatile als Markerstoffe angesehen werden, die zuverlässig einen Ökosystemzustand angeben können. Die Identifikation dieser Markerzustände und die Evaluation ihrer Nützlichkeit für spurenanalytische Anwendungen ist das Ziel dieser Arbeit. Es wurden drei Themen ausgewählt um die Verwendungsmöglichkeit des Insektengeruchssinns für Sensoranwendungen zu überprüfen: Fleischfrische, Todeszeitbestimmung (post mortem Intervall, PMI) und frühzeitige Feuererkennung. Spurenanalytik (GC-MS), Elektrophysiologie (EAG, GC-MS/EAD), Verhaltensversuche und Feldtests wurden angewandt, um die chemoökologische Interaktion der Schmeißfliege Calliphora vicina und der „Feuerkäfer“ Melanophila cuspidata, Merimna atrata and Acanthocnemus nigricans mit ihrer natürlichen Umgebung zu untersuchen. Die Resultate aus diesen Methoden erlaubten die Selektion von Volatilen, die drei Kriteria erfüllen: hohe Quantität, zuverlässige Emission und die ausschliessliche Emission von der untersuchten und keiner anderen Quelle. Diese drei Auswahlkriterien wurden aufgestellt um zu gewährleisten, dass die ausgewählten Markervolatile nicht nur zuverlässig mit dem physiologischen Status der Substrate (Alter von verderbendem Fleisch, PMI, Temperatur von erhitzten Spänen) korrelieren, sondern auch den Ansprüchen technischen Sensorsysteme genügen. Im Falle des alternden Fleisches unter warmen und trockenen Bedingungen ist Nonanal ein Zeiger für die korrekte Reifung des Fleisches zu Schinken. Dimethyl Trisulfid, Phenol und Indol zeigen das Verderben des Fleisches unter warmen und feuchten Bedingungen an. Bei niedrigen Temperaturen sind zunehmende 2,3 Butandiol Emission und abnehmende Nonanal Emission ein Zeiger für zunehmendes Verderben des Fleisches, sowohl unter trockenen als auch feuchten Bedingungen. Allerdings wurde 2,3 Butandiol nicht von C. vicina wahrgenommen, da die Fliege während der Vegetationszeit aktiv ist und unter diesen Bedingungen Dimethyl Trisulfid, Phenol und Indol emittiert werden. Nonanal, Hexanal, Dimethyl Disulfid, Dimethyl Trisulfid, Butan-1-ol und Phenol wurden als nützliche Volatile zur Eingrenzung der Leichenliegezeit ausgewählt. Die genannten Aldehyde sind typisch für die frühen Stadien der Verwesung, gefolgt von Butan-1-ol und den Schwefelsulfiden. Phenol wird hauptsächlich in den späteren Verwesungsstadien emittiert. Allerdings werden Phenol und 1-Butanol nicht von der Fliege wahrgenommen, da diese ein Generalist ist und nicht zwischen dem geblähten und aktivem Stadium der Verwesung unterscheiden muss. Daher wird zusätzlich die Geruchswahrnehmung des Speckkäfers Dermestes maculans betrachtet. Die Untersuchung anderer necrophager Insekten können weitere Einblicke in die Zeitabhängigkeit der Volatilemissionen während der Vertebratenverwesung geben. Die im Rahmen dieser Arbeit gewonnenen Ergebnisse erlauben eine Korrelation zwischen Emissionen und Leichenliegezeit und können eine Grundlage für eine volatilenbasierte post mortem Zeitbestimmung für Polizeikräfte sein. Terpene, aliphatische Aldehyde, Furfural und Methoxyphenole werden von erhitztem Holz emittiert. Komponenten aus all diesen Stoffklassen werden von M. cuspidata und M. atrata wahrgenommen, da diese Käfer den Erhitzungszustand von Baumstämmen nach einem Feuer erkennen können, um einen geeigneten Ovipositionsplatz zu finden. A. nigricans nimmt vor allem Methoxyphenole wahr, wohingegen M. atrata und M. acuminata Furfural zur Orientierung verwenden. Diese Unterschiede komplementieren das bekannte Verhalten dieser Käfer in ihrer natürlichen Umgebung, da A. nigricans auf verkohlten Stämmen, die große Mengen von Methoxyphenolen emittieren, seine Eier ablegt, und M. atrata und M. cuspidata auf Stämmen ihre Eier ablegen, die vom Feuer zwar äußerlich verkohlt, aber innerlich nur erhitzt wurden und daher Furfural emittieren. Halbleitergassensoren, die Furfural mit einer ausreichenden Selektivität vermessen können, können Holzbrände schon während ansteigender Temperatur und damit vor der Entzündung detektieren. Derartige Sensoren können sowohl als neuartige Frühwarnsysteme für die holzverarbeitende Industrie als auch für Waldbrandwarnsysteme oder den Haushalt dienen.

Frugivorous insects, specialised herbivores that consume fruit and seeds, are considered detrimental to host plant fitness. Their direct link to genetic fitness via consumption of plant reproductive tissue, and their negative socioeconomic association with agriculture exacerbates their harmful status. However, empirical testing of insect frugivore effects on host plants, and ecological research on the contribution of insect frugivores to multitrophic frugivory systems, is lacking. In the current study, direct effects of a non-mutualistic, insect frugivore/host plant system were tested and results showed variable effects. Beneficial, detrimental, but predominantly neutral effects on germination and seed production were observed between the Queensland fruit fly (Bactrocera tryoni) and tomato and capsicum plants. Significant effects on seed production were unexpected because infestation occurs after seed set. It was also found that eggplant, although a recorded host of B. tryoni, is inconsistent in its ability to sustain B. tryoni larvae through to its final instar. These results confirmed a simplification and presumption associated with insect frugivore (specifically fruit fly)/host plant interactions. Larval movement, infestation-induced fruit decay, pulp removal and germination were then investigated. For all hosts (tomato, apple and paw paw), treatments infested by B. tryoni decayed significantly quicker and to a greater extent than uninfested treatments, with obvious but variable changes to the texture and appearance. The movement of B. tryoni larvae, pattern of infestation-induced decay and pulp removal was unique and host dependent for all hosts. Only seeds from infested tomato were shown to germinate during the experiment. This indicated that host fruit characteristics are responsible, in part, for variable direct effects on host plant fitness by insect frugivores. Variable direct effects between insect frugivores and host plants, combined with the more rapid decay of infested fruits is likely to have implications for seed dispersal and seed predation by a third trophic level. The characteristics of fruit that are changed by infestation by an insect frugivore were then tested for their effect on a vertebrate frugivore, to illustrate the importance of recognising multitrophic interactions and indirect effects in frugivory. Specifically, seed predating rodents were incorporated into the study and their response to infested and uninfested fruits were recorded, as well as their reaction to the changes in fruit caused by insect frugivores (i.e. texture, smell, larvae presence and sound). Apple and pear infested with B. tryoni larvae were found to attract rodents, while infested tomato and paw paw had a neutral effect on the native rats. This differed from the predominant finding in the literature, which was a deterrent effect on avian seed dispersers. Vertebrate response to fruit infested with insect frugivores therefore, is variable. Assessing the indirect effect of insect frugivores on host plant fitness by attracting or deterring another trophic level requires knowledge of the direct effect between the introduced trophic level and the host plant. For example, the attraction of a seed predator may be as detrimental to host plant fitness as the deterrence of a seed disperser. This illustrates the complexity associated with assessing insect frugivore effects on host plant fitness. Results also indicated that differences in pulp texture, caused by infestation, have a significant effect on rodent preference for infested or uninfested treatments. Pulp texture is likely to effect rodent foraging efficiency, whereas the presence of B. tryoni larvae was observed to be inconsequential to rodent response to fruits. For rodents, and indeed any trophic level motivated by foraging efficiency, this finding raises the issue that for long lived fruiting plants, outside factors such as food abundance and competition for food, may cause a variable response to fruits infested by insect frugivores. From these investigations it has become apparent that insect frugivores are not consistently harmful to host plant fitness, as suggested by their negative stigma, but are likely to contribute variable effects, directly and indirectly, on multiple components of plant fitness and multitrophic frugivory systems.

During zebrafish gastrulation, highly coordinated cellular rearrangements lead to the formation of the three germ layers, ectoderm, mesoderm and endoderm. Recent studies have identified silberblick (slb/wnt11) as a key molecule that regulates gastrulation movement through a conserved pathway, which shares significant similarity with a signalling pathway that establishes epithelial planar cell polarity (PCP) in Drosophila (Heisenberg et al., 2000; Veeman et al., 2003), suggesting a role for cell polarity in regulating gastrulation movements. However, the cellular and molecular mechanisms by which slb/wnt11 functions during zebrafish gastrulation are still not fully understood. In the first part of the thesis, the three-dimensional movement and morphology of individual cells in living embryos during the course of gastrulation were recorded and analysed using high resolution confocal microscopy. It was shown that in slb/wnt11 mutant embryos, hypoblast cells within the forming germ ring display slower, less directed migratory movements at the onset of gastrulation, which are accompanied by defects in the orientation of cellular processes along the individual movement directions of these cells. The net movement direction of the cells is not changed, suggesting that slb/wnt11-mediated orientation of cellular processes serves to facilitate and stabilize cell movements during gastrulation. By using an in vitro reaggregation assay on mesendodermal cells, combined with an analysis of the endogenous expression levels and distribution of E-cadherin in zebrafish embryos at the onset of gastrulation, E-cadherin mediated adhesion was found to be a downstream mechanism regulating slb/wnt11 function during gastrulation. Interestingly, the effects of slb/wnt11 on cell adhesion appear to be dependent on Rab5-mediated endocytosis, suggesting endocytic turnover of cell-cell contacts as one possible mechanism through which slb/wnt11 mediates its effects on gastrulation movements. - Die Druckexemplare enthalten jeweils eine CD-ROM als Anlagenteil: QuickTimeMovies (ca. 23 MB)- Übersicht über Inhalte siehe Dissertation S. 92 - 93"

A plethora of naturally-produced steroid hormones, or artificial homologues of them, are being introduced into the aquatic and terrestrial environments each year. Two examples of these are the natural oestrogen 17-oestradiol (E2) and the oestrogen receptor antagonist, Bisphenol A (BPA), both of which target the ribonucleoprotein telomerase through upregulation of its telomerase reverse transcriptase component, TERT. The main objectives of this study were firstly to isolate and characterize the actual mRNA sequence for the telomerase catalytic subuninit, Tert, in rainbow trout (Oncorhynchus mykiss) (Walbaum, 1792) and European purple sea urchin (Paracentrotus lividus) (Lamarck, 1816), with the aim of developing qPCR assays for the amplification and quantification of Tert. Further objectives were to use these assays in controlled exposure studies to establish whether and to what extent the aforementioned chemicals regulate Tert transcription and by doing so further understand the mechanism of Telomerase gene expression and the extent to which environmental oestrogen can interfere. The initial step of sequence characterization and assay devlopment was successful in the case of rainbow trout where two possible splice variants of Tert mRNA are identified, omTertShort and omTertLong. Two qPCR assays were developed for the relative quantification of both of these splice variants in rainbow trout samples, the latter of these successfully amplifying its target in test samples. In order to demonstrate in vitro and in vivo modulation of telomerase activity and mRNA expression, early life-stages of rainbow trout and purple sea urchin, as well as rainbow trout hepatocytes, were exposed to a range of concentrations of E2 and BPA. Purple sea urchin embryos were exposed to 200, 20 and 2 ng E2/ml for 28 hours until they had reached the stage of pluteus larvaes. Rainbow trout embryos were exposed to 500, 20 and 0.1 ng E2/ml and 600 and 150 ng BPA/ml for 167 days from immediately after fertilization. Rainbow trout hepatocytes were exposed to 20 and 2 ng E2/ml for 48 hours. The results from this study show that telomerase activity as well as TERT mRNA expression can be significantly modulated by exposure to oestrogens and other oestrogenic chemicals. E2 concentrations as low as 20 ng/ml lead to an increase in telomerase activity early-life stages of purple sea urchin and upregulation in the transcription of Tert mRNA in unhatched rainbow trout embryos. BPA induced similar response (600 ng/ml) in hatched rainbow trout alevins larvae. Very high exposures to E2 (500 ng/ml) do however lead to downregulation of Tert mRNA in hatched alevins larvae. Differential regulatory response can be observed between different tissue types of 167 day old fry, with an upregulatory response observed at 0.1 ng E2/ml in liver and muscle tissues, but not in brain. Similarly, brain tissues were observed expressing significantly less mRNA than liver and muscle samples when exposed to BPA (150 ng/ml). It is evident that the previously observed link between environmental oestrogens and telomerase is also present in the two test species examined; purple sea urchin and rainbow trout.

Visual pigments are light sensitive receptors in the eye that form the basis of sensory visual transduction. This thesis presents three studies that explore visual pigment proteins in vertebrates using a number of computational and experimental methods in an evolutionary framework. The objective is not only to identify, but also to experimentally investigate the functional consequences of genetic variation in vertebrate visual pigments. The focus is on great bowerbirds (Chlamydera nuchalis), which are a model system in visual ecology due to their spectacular behaviour of building and decorating courtship bowers. There are 4 chapters: Chapter 1 introduces background information on visual pigments and vision in birds. Among visual pigment types, the short-wavelength-sensitive (SWS1) pigments have garnered particular interest due to the broad spectral range among vertebrates and the importance of UV signals in communication. Chapter 2 investigates the evolutionary history of SWS1 in vertebrates with a view toward its utility as a phylogenetic marker. Chapter 3 investigates SWS1 evolution and short-wavelength vision in birds, with particular focus on C. nuchalis and its SWS1. The evolution of spectral tuning mechanisms mediating UV/violet vision in passerines and parrots is elucidated in this chapter using site-directed mutagenesis, protein expression, and phylogenetic recreation of ancestral opsins. While cone opsins mediate colour vision in bright light, the rhodopsin visual pigment contained in rod photoreceptors is critical for dim light vision. Detailed characterization of rhodopsin function has only been conducted on a few model systems. Chapter 4 examines C. nuchalis RH1 using a number of functional assays in addition to absorbance spectra, including hydroxylamine sensitivity and the rate of retinal release. This chapter includes an investigation into the role of amino acid mutations typical of dim-light adapted vertebrates, D83N and A292S, in regulating functional properties of bovine and avian RH1s using site-directed mutagenesis. Together these chapters describe naturally occurring mutations in visual pigments and explore the way they can influence visual perception. These represent one of the few investigations of visual pigments from a species that is not a model lab organism and form a significant contribution to the field of visual pigment biochemistry and evolution.

Aerial control using 1080 (sodium monofluoroacetate) baits is widely used in New Zealand for the control of introduced brushtail possums (Trichosurus vulpecula), with the aim of protecting national conservation and agricultural values from these damaging pests. This thesis integrates research, completed over 25 years, that was motivated by growing recognition in the 1970s of the extent of possum impacts and the need to improve the effectiveness and efficiency of the control operation. Field research assessed the palatability of three types of cereal-based pellet baits and carrot baits in different regions, habitat types and seasons. Palatability was assessed by the consumption of the different bait types presented independently of each other on 15-30 plots, with rotation of bait types at plots on successive nights to provide equal exposure to each bait type. There was regional variation in possums' bait preferences, possibly reflecting genotypic differences, whereas seasonal variation was less evident. Carrot bait was preferred or equally preferred to cereal bait in 14 out of 20 field trials. The proportion of possums eating baits was then investigated by, firstly, developing a technique for tracing bait acceptance using rhodamine B, a UV-fluorescent dye. In four field trials, more than 95% of possums accepted three types of dye-marked bait, eliminating bait refusal as a major reason for low kills in winter control operations. In a fifth trial, conducted in summer, only 68% of possums accepted bait suggesting that seasonal availability of favoured foods may influence bait acceptance. Since possums must encounter baits before deciding whether to eat them, field studies were undertaken to assess the coverage achieved in normal aerial baiting operations. Large gaps, up to 400 m in width, were often found between baiting swaths; these could allow some possums to survive. A controlled field experiment, using acceptance of rhodamine-dyed bait as a measure of effectiveness, showed that bait distribution was least accurate where flight paths were not marked. Where gaps of 100 m between flight paths were deliberately created, bait acceptance was slower and less than where coverage was complete. Sowing baits at 3 kg/ha was as effective as at 10 kg/ha, indicating the potential for substantially reducing operational costs by using machinery capable of faultlessly distributing baits at low rates. Navigational guidance systems were evaluated and found to improve the accuracy of bait distribution. During 1993-1997, when a lower sowing rate of 5 kg/ha was adopted operationally by regional managers, control effectiveness was unchanged but annual savings of around $9 million accrued. Because of the lack of suitable sowing machinery, a bucket was developed to permit faultless distribution of baits at lower rates, demonstrating the possibility of yet further cost-savings. The possibility of seasonal food availability affecting bait acceptance was investigated in three different forest habitats. Dyed baits were aerially distributed on 100 ha at each site in each season over two years. In each trial, fat-based condition indices of possums were calculated and the abundance of possum-preferred plant foods described. Bait acceptance was consistently high (85-100%) in the 24 trials, and was not influenced by either condition or availability of preferred foods. It seems likely that seasonal variation in operational effectiveness is caused by either the availability of sharply seasonal, scarce foods that possums may feed on intensively for brief periods, or by warmer temperatures that render 1080 less effective. The influence of 1080 on acceptance of (rhodamine-dyed) baits was investigated in a field trial. Examination of possums for dye-marking showed that 25% of possums refused to eat either a lethal quantity of bait or any bait at all, compared with 98% of possums eating non-toxic bait. This indicated that 1080 is aversive to possums, which is a potential major reason for their surviving control operations. Pen trials were therefore conducted to further examine the problem and to seek solutions. Toxic carrot baits were rejected by 27.5% of possums, equally by smell and taste aversion, whereas toxic cereal pellets were rejected by 34%, mainly by taste aversion. Orange and cinnamon were shown to be among the most preferred of 42 flavours tested and, when applied to toxic baits, 1080 was effectively masked. Bait refusal was reduced to ≤7%, the same as that recorded for possums presented with flavoured non-toxic baits. For long-term control of possum populations, aerial 1080 baiting can be used sequentially with other poisoning methods. However, the compatibility of these methods is dependent on the likelihood of possums developing bait shyness if sublethally dosed. Studies were therefore conducted to characterise and compare the four main toxicants used (1080, cyanide, cholecalciferol and brodifacoum) for induction and mitigation of bait shyness. Shyness was induced in approximately 80% of possums sublethally dosed with cyanide, 60% with 1080, 20% with cholecalciferol, and 0% with brodifacoum. Cyanide and 1080 shyness were found to persist in many possums for at least 12 and 24 months, respectively. Use of alternative bait types, and of baits containing an alternative slow-acting toxin (brodifacoum) were shown to be effective ways of overcoming shyness. This, and other related research, is reviewed to provide operational specifications that maximise the likelihood that all targeted possums will (i) encounter bait, (ii) eat it, and (iii) die. The likely future use of aerial 1080 baiting is described and the technological, economic, environmental and social constraints on its sustainability are discussed. Finally, the uptake of the research by possum managers is considered, and areas identified in the thesis where information is incomplete are summarised as prioritised topics for further research.

L'étude des mécanismes endocriniens est particulièrement intéressante du fait du rôle majeur des hormones dans la régulation des interactions entre la physiologie d'un organisme, son comportement, et les modifications de son environnement. Cette thèse s'est intéressée aux relations entre le statut hormonal, les performances de reproduction et le succès reproducteur d'un oiseau marin longévif, le manchot Adélie Pygoscelis adeliae, dans un contexte environnemental soumis à des changements. Le statut endocrinien de manchots mâles a été manipulé en utilisant des implants dégradables sous-cutanés diffusant l'hormone d'intérêt ou un inhibiteur de sa sécrétion. Les effets d'une modification des niveaux d'hormones sur l'investissement parental pendant l'incubation ont été mesurés à l'aide d'observations directes et d'oeufsfactices enregistrant les paramètres d'incubation. Les niveaux de corticostérone - hormone dite de stress, de prolactine - hormone des soins parentaux, et de testostérone - hormone liée aux comportements sexuels et à l'agressivité, ont été manipulés. Les effets d'une augmentation des niveaux de corticostérone sur les performances et le succès reproducteur pendant la période de l'élevage des poussins ont également été mesurés. Enfin, les conséquences d'une légère élévation des niveaux de corticostérone pendant l'ensemble de la saison de reproduction en termes de comportement et de succès reproducteur ont été examinées. Une augmentation des niveaux de corticostérone a globalement diminué les performances et le succès de reproduction. D'autre part, une modification des niveaux de prolactine ou de testostérone a affecté la durée etles paramètres d'incubation, suggérant une implication de ces deux hormones dans le contrôle de la phénologie de la reproduction. Les résultats présentés dans cette thèse mettent l'accent sur le fait que la relation entre statut endocrinien et performances de reproduction est dose, état et contexte dépendante. Nos résultats illustrent le rôle majeur des hormones étudiées dans la régulation de l'effort reproducteur, et soulignent également l'importance de considérer les interactions entre les organismes et leur environnement.

Visual pigments are light sensitive receptors in the eye that form the basis of sensory visual transduction. This thesis presents three studies that explore visual pigment proteins in vertebrates using a number of computational and experimental methods in an evolutionary framework. The objective is not only to identify, but also to experimentally investigate the functional consequences of genetic variation in vertebrate visual pigments. The focus is on great bowerbirds (Chlamydera nuchalis), which are a model system in visual ecology due to their spectacular behaviour of building and decorating courtship bowers. There are 4 chapters: Chapter 1 introduces background information on visual pigments and vision in birds. Among visual pigment types, the short-wavelength-sensitive (SWS1) pigments have garnered particular interest due to the broad spectral range among vertebrates and the importance of UV signals in communication. Chapter 2 investigates the evolutionary history of SWS1 in vertebrates with a view toward its utility as a phylogenetic marker. Chapter 3 investigates SWS1 evolution and short-wavelength vision in birds, with particular focus on C. nuchalis and its SWS1. The evolution of spectral tuning mechanisms mediating UV/violet vision in passerines and parrots is elucidated in this chapter using site-directed mutagenesis, protein expression, and phylogenetic recreation of ancestral opsins. While cone opsins mediate colour vision in bright light, the rhodopsin visual pigment contained in rod photoreceptors is critical for dim light vision. Detailed characterization of rhodopsin function has only been conducted on a few model systems. Chapter 4 examines C. nuchalis RH1 using a number of functional assays in addition to absorbance spectra, including hydroxylamine sensitivity and the rate of retinal release. This chapter includes an investigation into the role of amino acid mutations typical of dim-light adapted vertebrates, D83N and A292S, in regulating functional properties of bovine and avian RH1s using site-directed mutagenesis. Together these chapters describe naturally occurring mutations in visual pigments and explore the way they can influence visual perception. These represent one of the few investigations of visual pigments from a species that is not a model lab organism and form a significant contribution to the field of visual pigment biochemistry and evolution.

<p>Management of predation on sea turtle nesting beaches is vital to conservation efforts for the vulnerable loggerhead turtles (<i>Caretta caretta</i>) and olive ridley turtles (<i>Lepidochelys olivacea</i>). Sea turtles increasingly face threats from invasive and human-tolerant mammalian predators as human disturbances on nesting beaches rises. The intensity of mammalian predation has increased in Las Baulas National Park in Costa Rica which is an important nesting site for several species of threatened and endangered sea turtles. I analyzed loggerhead and olive ridley nest predation on four beaches in the United States and Costa Rica that were chosen for variations in degree of human disturbance and management strategies. My objectives were to 1) determine if egg predation rates differ at the four sites, 2) determine the most destructive predators at each location, and 3) suggest management options to alleviate mammalian threats to turtle clutches on Playa Grande and Playa Cabuyal in Costa Rica. My results show that the beaches without a nest protection or predator control program had very high rates of predation. Invasive mammalian predators and mammalian predators associated with human disturbance were the most destructive at the four sites. I recommend that regulations regarding dogs and the take of eggs from the beach are enforced at Playa Cabuyal and that physical nest protection is rapidly implemented at Playa Grande. I also recommend that the National Park consider managing raccoon predation by removing problem individuals, but caution that they do so in a way that maintains the animals’ role in the ecosystem.</p>