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Holography has the ability to render three-dimensional information of a recorded scene by capturing both the amplitude and phase of light incident on the recording medium. The application of digital camera technology and high-speed computing means digital holograms can be analysed numerically and novel applications can be found for this technology. This thesis explores the potential for both inline and off-axis digital holographic microscopy to study the three-dimensional swimming behaviour of bacteria. A high-magnification (225x) digital holographic microscope was designed and constructed with the ability to switch easily between inline and off-axis imaging modalities. Hardware aspects, in particular the illumination source, the choice of camera and data transfer rates, were considered. Novel strategies for off-axis holography combining dark field microscopy were designed and implemented. The localisation accuracy of the inline imaging modality was assessed by studying samples of polystyrene microspheres. The microscope is sensitive to stage drift on the order of angstroms per second and can successfully localise microspheres in dilute suspensions at least 100μm from the objective specimen plane. As a simple test of the capabilities of the microscope, the diffusion coefficient of a 0.5μm microsphere was found to be isotropic and consistent with the theoretical value. Amplitude and phase image reconstructions from the off-axis modality are demonstrated. High-magnification dark field off-axis holographic microscopy is shown to be superior to inline microscopy in localising 100nm gold nanoparticles. An artifact from our method of dark-field imaging, however, restricts the depth range to 15μm. A lower-magnification (45x) configuration of the microscope was used to study the 3D swimming behaviour of wild type Escherichia coli as a qualitative demonstration of the potential for this instrument in microbiological applications.

Nos últimos anos, a preparação de substâncias enantiomericamente puras tornou-se uma preocupação constante e um desafio para a indústria. Tal fato é consequência da relação tênue entre a estrutura espacial das substâncias e os diferentes sabores, odores, processos farmacocinéticos e farmacodinâmicos e toxicidade que elas desempenham no organismo. Nesse contexto, a síntese assimétrica de compostos carbonílicos ?-alquil substituídos opticamente ativos tem recebido grande atenção já que estes são importantes blocos de construção na síntese de fármacos quirais e produtos naturais. Assim, o presente trabalho buscou o desenvolvimento de metodologias que permitissem a preparação desses compostos enantiomericamente puros através do controle na formação de centros estereogênicos usando a biocatálise. Para isso, foram preparadas diversas olefinas ativadas por grupos retiradores de elétrons (carbonila) utilizando dois procedimentos, a saber, reações de condensação aldólica e esterificação. Esses compostos carbonílicos ?,?-insaturados foram obtidos com bons rendimentos (48-94%) e foram submetidos à reduções biocatalíticas mediadas por células íntegras de micro-organismos. Inicialmente, utilizou-se a olefina (E)-4-fenil-3-metil-3-buten-2-ona (51a) como substrato modelo para a otimização das condições experimentais e determinação daquela que fornecesse o produto biorreduzido com a melhor conversão e pureza enantiomérica. Nesses testes, avaliaram-se diversos parâmetros reacionais, tais como, composição do meio reacional (aquoso, bifásico e tamponante), presença de aditivos (glicose e catalisador de transferência de fase), diferentes micro-organismos e efeito do uso de resina adsorvente Amberlite XAD-7. Dessa forma, a biorredução em meio aquoso, sem aditivos, mediada pela levedura LSC-II e com a utilização da resina Amberlite XAD-7 foi a condição reacional que apresentou o melhor excesso enantiomérico (81%) e foi a selecionada para um estudo metodológico que avaliou os efeitos estérico e eletrônico dos substituintes. Observou-se que a posição do substituinte no ?-aril, o alongamento do ?-substituinte e a adição de um grupamento éster apresentaram um importante papel na estereosseletividade da reação. Por fim, avaliou-se a aplicação da metodologia na síntese enantiosseletiva da muguesia, uma fragrância floral. Os resultados obtidos revelaram o grande potencial da metodologia na biorredução quimiosseletiva de compostos carbonílicos ?,?-insaturados e preparação de substâncias opticamente ativas. / In recent years, the preparation of enantiomerically pure compounds has become a concern and a challenge to the industry. This fact is a consequence of the tenuous relationship between the spatial structure of substances and the different flavors, odors, pharmacokinetic and pharmacodynamic processes and toxicity that they play in the body. In this context, the asymmetric synthesis of ?-alkyl substituted optically active carbonyl compounds has received great attention because they are important building blocks in the synthesis of chiral drugs and natural products. Thus, this study aimed to develop methodologies that allow the preparation of this enantiomerically pure compounds by controlling the formation of stereogenic centers using biocatalysis. To this end, various activated olefins substituted with electron withdrawing groups (carbonyl) were synthesized using two procedures, aldol condensation and esterification reactions. These ?,?-unsaturated carbonyl compounds were obtained in good yields (48-94%) and subjected to the biocatalytic reduction mediated by whole cells of microorganisms. Initially, we used the olefin (E)-3-methyl-4-phenyl-3-buten-2-one (51a) as a model substrate for the optimization of the experimental conditions and determine wich one provide the product with better conversion and enantiomeric purity. In these tests, we evaluated various reaction parameters such as the composition of the reaction medium (aqueous, biphasic and buffer), the presence of additives (glucose and phase transfer catalyst), different microorganisms and adsorbing effect of using Amberlite XAD-7. Thus, the biorreduction in aqueous medium, without additives, mediated by LSC-II yeast and with the application of Amberlite XAD-7 was the reaction condition that presented the best enantiomeric excess (81%) and was selected for a methodological study that evaluated the steric and electronic effects of the substituents. It was observed that the substituent in the ?-position of the aryl group, the size increase of the ?-substituent and the presence of an ester group had an important role in the stereoselectivity of the reaction. Finally, application of the methodology on muguesia enantioselective synthesis, a floral fragrance, was evaluated. The results obtained showed the high potential of the methodology in chemoselective bioreduction of ?,?-unsaturated carbonyl compounds and preparation of optically active substances.

Potential vegetative and reproductive increases in yield, as well as the biological efficacy against Fusarium verticillioides and F. proliferatum causing ear and stem rot in maize crops of commercially-formulated micro-organism formulation T-Gro (Trichoderma harzianum isolate DB103 WP) combined with Spartacus (Beauveria bassiana isolate DB 105 WP), T-Gro combined with Armenius (Bacillus subtilis isolate DB 109 WP), T-Gro combined with Maximus (Bacillus subtilis isolate DB 108 WP), T-Gro combined with Shelter (Bacillus subtilis isolate DB 101), T-Gro combined with Bismarck (Microbacterium maritypicum isolate DB 107 WP), as well as individual treatments of T-Gro, Armenius, Bismarck, Maximus and Shelter, were investigated when applied to maize seed and soil under field conditions. All the micro-organism treatments were compared with Thiram 750WP (750g/kg thiram WP) and an untreated control. The micro-organism treatments showed an increase in vegetative as well as reproductive yields when compared to the reference product Thiram 750 WP and the untreated control. There were no observations of adverse effects on the germination of maize seed in all the treatments that were applied. The three isolates B. subtilis, T. harzianum, and M. maritypicum, showed a significant reduction in vascular tissue discolouration of the main and ear stems, indicating a potential to be used in the reduction and control of diseases caused by Fusarium spp. Results also showed poor to very good increases of stem and foliage biomass as well as cob yield per plant produced by the micro-organism treatments when compared to the untreated control. The highest cob yield per plant that differed significantly from the untreated control was produced by T-Gro and Shelter. No phytotoxicity of any kind was observed with the application of the micro-organism formulations and they could therefore be deemed suitable to be used for the treatment of maize seed. The micro-organism formulations containing fungal and bacterial biological control agents have the potential to be used in commercial maize production to increase vegetative and reproductive yields and reduce the severity of ear and stem rot in maize. / Agriculture Animal Health & Human Ecology / M.Sc. (Agriculture)

Background There is a transition towards minimally invasive restorative techniques in restorative dentistry based upon reducing bacterial viability and encouraging remineralization of caries infected tissue. To improve the predictability of the antibacterial and remineralization potential of carious dentine by either the application of medicaments or placement of restorative materials that encourage remineralization would be a significant benefit in disease management. Materials and Methods An experimental model was developed using a chemostat for in vitro analysis of the effects of silver fluoride followed by potassium iodide (AgF/KI) and ozone treatment on non demineralized and demineralized dentine. Electron Probe Micro Analysis (EPMA) and Scanning Electron Microscopy (SEM) on the treated dentine were conducted to investigate ion transfer, and biofilm formation. Bacteria growth was measured by optical density. An in vitro caries model using a chemostat was developed to determine the ability of glass ionomer cement and composite resin to inhibit dentinal degradation in adjacent dentine and to measure ion exchange at the restorative interface. Tests were made to determine the bond strength between dentine and glass ionomer cement after application of silver fluoride to the surface of the dentine. Results S. mutans migrated through all dentine samples. Samples treated with AgF/KI had significantly lower optical densities than the corresponding controls. Optical density readings were significantly lower in demineralized dentine treated with AgF/KI than non demineralized dentine. There were lower but not significant differences in the optical density readings between ozonated and non ozonated dentine. An S. mutans biofilm covered all control discs. No biofilm was detected on discs treated with AgF/KI and these discs were significantly more resistant to further demineralization than the control discs. Detectable amounts of silver and fluoride were found up to 450 μm in the AgF and AgF/KI sections. Ozone infusion prevented S. Mutans and L. acidophilus biofilm formation on all the treated dentine samples, biofilm was present on all control specimens. There was calcium and phosphorus present in all auto cure glass ionomer cements to a depth beyond 50 microns. Aluminium and strontium ions were also present in dentine except strontium subjacent to Ketac Molar restorations. Fluoride uptake was significantly higher under glass ionomer cement restorations where the dentine was pretreated with AgF/KI compared to non treated specimens. Silver and iodine deposits were present in demineralized dentine treated with AgF/KI. Calcium and phosphorus levels up to 130 microns from the restorative interface were similar to non demineralized dentine adjacent to auto cure glass ionomer cements and half that adjacent to composite resin. There was significant surface degradation in auto cure glass ionomer cements compared to composite resin. Washing away the AgF/KI precipitate produced higher bond strengths to dentine than samples where the precipitate remained. Conclusions Under the conditions of these in vitro studies, the application of AgF/KI and ozone pharmacologically reduces the initiation and rate of dentine caries. Glass ionomer cements were shown to protect dentine from experimental carious degradation and assist with remineralization. AgF/KI application enhances remineralization beneath glass ionomers and does not interfere with bond strengths. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1344616 / Thesis (Ph.D.) - University of Adelaide, Dental School, 2008

Potential vegetative and reproductive increases in yield, as well as the biological efficacy against Fusarium verticillioides and F. proliferatum causing ear and stem rot in maize crops of commercially-formulated micro-organism formulation T-Gro (Trichoderma harzianum isolate DB103 WP) combined with Spartacus (Beauveria bassiana isolate DB 105 WP), T-Gro combined with Armenius (Bacillus subtilis isolate DB 109 WP), T-Gro combined with Maximus (Bacillus subtilis isolate DB 108 WP), T-Gro combined with Shelter (Bacillus subtilis isolate DB 101), T-Gro combined with Bismarck (Microbacterium maritypicum isolate DB 107 WP), as well as individual treatments of T-Gro, Armenius, Bismarck, Maximus and Shelter, were investigated when applied to maize seed and soil under field conditions. All the micro-organism treatments were compared with Thiram 750WP (750g/kg thiram WP) and an untreated control. The micro-organism treatments showed an increase in vegetative as well as reproductive yields when compared to the reference product Thiram 750 WP and the untreated control. There were no observations of adverse effects on the germination of maize seed in all the treatments that were applied. The three isolates B. subtilis, T. harzianum, and M. maritypicum, showed a significant reduction in vascular tissue discolouration of the main and ear stems, indicating a potential to be used in the reduction and control of diseases caused by Fusarium spp. Results also showed poor to very good increases of stem and foliage biomass as well as cob yield per plant produced by the micro-organism treatments when compared to the untreated control. The highest cob yield per plant that differed significantly from the untreated control was produced by T-Gro and Shelter. No phytotoxicity of any kind was observed with the application of the micro-organism formulations and they could therefore be deemed suitable to be used for the treatment of maize seed. The micro-organism formulations containing fungal and bacterial biological control agents have the potential to be used in commercial maize production to increase vegetative and reproductive yields and reduce the severity of ear and stem rot in maize. / Agriculture Animal Health and Human Ecology / M.Sc. (Agriculture)

Le procédé de friture utilise les huiles usagées comme fluide de chauffe pour sécher les boues humides par contact direct tout en incorporant une fraction. Le produit obtenu est un combustible solide à fort pouvoir calorifique, hygiènisé, stockable et transportable sans risque. Une étude expérimentale et théorique a permis d'apporter les éléments nécessaires à la conception du procédé, tant sur le plan énergétique que sur les plans environnemental et sanitaire. Un modèle de cinétique de friture, est proposé pour le séchage et la dynamique d'absorption d'huile, basé sur une loi de diffusion de l'eau dans la boue et un remplacement de l'eau par l'huile volume pour volume. Le modèle est ajusté aux données expérimentales obtenues par méthode gravimétrique sur des cylindres de boues (4-12 mm). Les températures employées varient entre 110°C et 140°C. Le temps de séchage, compris entre 10 et 20 min est réalisable dans une unité de séchage en continu. Un bilan d'énergie a permis de déterminer l'évolution du coefficient de transfert convectif en mettant en évidence trois phases successives. Une corrélation adimensionnelle est proposée, regroupe le flux évaporatoire normé, les caractéristiques thermiques d'huile et la géométrie de la boue. Une étude de la cinétique de destruction thermique des micro-organismes pathogènes a permis de construire un modèle estimant le temps de friture nécessaire pour hygiéniser une boue dont la température est connue dans le temps. Les conditions de friture sous vide à une température d'ébullition de 80°C ont été étudiées et comparées à d'autres procédés de séchage. Enfin, pour évaluer l'insertion du procédé dans une filière de valorisation énergétique par combustion, une étude expérimentale des émissions atmosphériques a été réalisée pour une combustion en lit fixe (850°C). Une attention particulière a été accordée aux émissions des hydrocarbures aromatiques polycycliques vu leurs impacts cancérigènes pour la santé humaine et leur toxicité pour les écosystèmes. Les résultats ont montrés que les émissions de boues frites sont d'environ 5000 microgramme/gramme soit 10 fois plus importantes que celles du bois et 10 fois moins importantes que celles des huiles usagées. / Fry-drying is an alternative for heat and mass transfer intensification. The process re-uses waste oil as a heating medium for drying by contact with the wet sludge. The fry-dried product is a granular solid fuel with high heating value. The product is sterile, stored, and transported safely. Experimental procedure and theoretical investigation have to provide basic information for the process design, in terms of energy and environment. A kinetic model of frying is proposed for water loss and oil intake. The model has been fitted to the data obtained by a weighing method on sludge cylinders (4-12 mm diameter) at an oil temperature ranging from 110°C to 140°C. The drying time between 10 and 20 min is effective in a continuous dryer. The global energy balance has enabled the calculation of a convective heat transfer coefficient. The results show that convection is boiling flow dependent. The study suggests a dimensionless correlation involving the normalized boiling flux, thermal oil property, and the sludge size. An estimation of kinetics of thermal destruction of pathogens during vacuum frying (80°C boiling point) was made. The calculations were compared with the current thermal dryer performance. Finally, to evaluate the integration of the process in a sequence of energy recovery by combustion, an experimental study of air emissions was achieved for fixed-bed combustion (850°C). Particularly, the chemical characterization was focused on the 16 Polycyclic Aromatic Hydrocarbons a group of highly classified carcinogens. The results showed that emissions of fry-dried sludge are approximately 5000 microgramm/gramm thus 10 times higher than those of wood and 10 times smaller than those of oils.

Les populations naturelles sont confrontées à des changements environnementaux. Pour y faire face, différentes réponses ont été sélectionnées au cours de l'évolution. Parmi elles se trouvent la plasticité phénotypique et l'adaptation génétique. Etudier les liens existants entre elles est une manière de comprendre les dynamiques des populations et de prévoir leurs réponses à un environnement changeant. Dans la présente étude, je me suis attaché à étudier ces liens à plusieurs échelles (intra- et interspécifique), chez le complexe d'espèces cryptiques de la micro-algue Alexandrium minutum, et ce à la fois in vitro et in situ. En ce qui concerne la plasticité phénotypique, ces deux espèces proches montrent de profondes différences, soulignant les liens entre divergence génétique et écologique. Au niveau intraspécifique, il apparaît que face à des variations de facteurs abiotiques, les populations ajustent les niveaux d'expression de certains gènes (notamment impliqués dans des fonctions de motilité et d'interactions intercellulaires dans des environnements froids à faible salinité). D'autre part, les populations montrent de la différentiation génétique à la fois à faible échelle spatiale, au cours du temps, et lorsque la communauté change. Pour conclure, il existe une interaction directe entre divergence génétique et changements d'expression de gènes. En plus de poser de nombreuses questions quant aux capacités de réponse des populations, ces résultats soulignent comment plasticité phénotypique et changements génétique sont liés et interagissent. Ils offrent une perspective nouvelle sur les mécanismes qui sous-tendent les réponses des populations à leur environnement. / Natural populations face environmental changes. In this context, different responses were evolutionnary selected. Among them are phenotypic plasticity and genetic adaptation. Studying the links between these two types of response is a way to understand population dynamics and to predict how they may respond to a changing environment. In the present Ph.D thesis, I focused on studying these links at several scales (intra- and interspecific), in the cryptic species complex of the microalga Alexandrium minutum, both in vitro and in situ. With respect to phenotypic plasticity, these two closely related species show profound differences, highlighting the links between genetic and ecological divergence. At the intraspecific level, it appears that, when facing abiotic factors variations, populations adjust the expression levels of certain genes (notably involved in motility related functions and intercellular interactions under low-salinity and cold environments). On the other hand, populations show genetic differentiation at both small spatial scale, over time, and when the community changes. To conclude, there is a direct interaction between genetic divergence and changes in gene expression. In addition to asking many questions about the response capabilities of populations, these results highlight how phenotypic plasticity and genetic changes are linked and interact. They offer new perspectives on the mechanisms underlying population responses to their environment.

<p>The objective of the dissertation was to study the cabin environment, and identify personal and environmental risk factors, associated with symptoms, and perception of cabin air quality. Another objective was to study if ban of smoking, and increased relative air humidity on intercontinental flights, could have a beneficial health effect. The studies were performed among Scandinavian cabin crew in one Airline Company. Office workers from the same company served as controls. Exposure differed between cruise and non-cruise conditions. Air humidity was very low during intercontinental flights (3-8%). Concentration of moulds, bacteria, formaldehyde, and ozone was low. Tobacco smoking increased respirable particles in the cabin air, from 3 to 49 mg/m3, and increased cotinine in urine. The ETS-exposure was highest in the aft part of the cabin. Symptoms and environmental complaints were more common among flight crew than office workers. We could identify personal factors of importance, and certain conditions that could be improved, to achieve a better cabin environment. There was an association between symptoms and environmental perceptions and work stress, lack of influence on working condition, and a history of atopy. After ban on smoking in aircraft, there was a decrease of ocular and general symptoms, and increased tear-film stability in aircrew. Air humidification reduced headache and ocular, nasal, and dermal dryness symptoms, increased tear-film stability, and increased nasal patency. Our result indicates that ETS and low air humidity are important environmental factors in aircraft, and that atopy, and work stress could be significant risk factors for symptoms and environmental perceptions.</p>