Tesi sul tema "Algae Physiology"

Les microalgues ont développé des approches distinctes pour moduler l'absorption de la lumière et son utilisation par leurs photosystèmes en réponse à des stimuli environnementaux. Dans ce rapport de Thèse je présente les différentes stratégies employées par une algue d'eau douce (Chlamydomonas reinhardtii) et une algue marine (Phaeodactylum tricornutum) pour optimiser leur acclimatation à l'environnement.Dans la première partie de ce rapport, je propose un modèle de cellules entières de la diatomée marine Phaeodactylum tricornutum obtenue par analyses spectroscopiques et biochimiques ainsi que par l’obtention d’images par microscopie électronique et reconstitution 3-D. Ce modèle a été utilisé pour répondre aux questions suivantes i. comment est structuré un chloroplaste secondaire pour faciliter les échanges avec le cytosol à travers les quatre membranes qui le délimitent ii. comment sont structurées les membranes photosynthétiques afin d’optimiser l'absorption de lumière et le flux d'électrons et iii. comment les chloroplastes et les mitochondries sont organisés pour optimiser l'assimilation du CO2 par échange ATP / NADPH.La deuxième partie de ce rapport porte sur la régulation de la collection de la lumière et de sa dissipation chez Chlamydomonas grâce à l'étude d'une part du rôle de la perception de la couleur de la lumière et d'autre part du métabolisme sur la dissipation de l'excès de lumière par quenching non photochimique (NPQ). En utilisant des approches biochimiques et spectroscopiques, j'ai mis en évidence un lien moléculaire entre la photoréception, la photosynthèse et la photoprotection chez Chlamydomonas via le rôle du photorécepteur phototropine, démontrant ainsi que le métabolisme, en plus de la lumière, peut aussi affecter ce processus d'acclimatation.En conclusion, ce travail de thèse révèle l'existence et l'intégration des différentes voies de signalisation dans la régulation des réponses photoprotectrices mises en place chez les microalgues marines et d'eau douce<br>Microalgae have developed distinct approaches to modulate light absorption and utilization by their photosystems in response to environmental stimuli. In this Ph.D Thesis, I characterised different strategies employed by freshwater (Chlamydomonas reinhardtii) and marine algae (Phaeodactylum tricornutum) to optimise their acclimation to the environment.In the first part of this work, I used spectroscopic, biochemical, electron microscopy analysis and 3-dimentional reconstitution to generate a model of the entire cell of the marine diatom Phaeodactylum tricornutum. This model has been used to address the following questions: i. how is a secondary chloroplast structured to facilitate exchanges with the cytosol via its four membranes envelope barrier ii. how have diatoms shaped their photosynthetic membranes to optimise light absorption and downstream electron flow and iii. how the cellular organelles interact to optimise CO2 assimilation via ATP/NADPH exchanges.In the second part, I have focused on the regulation of light harvesting and dissipation in Chlamydomonas by studying the role of perception of light colour and metabolism on excess light dissipation via the Non-Photochemical Quenching of energy (NPQ). Using biochemical and spectroscopic approaches, I found a molecular link between photoreception, photosynthesis and photoprotection in Chlamydomonas via the role of the photoreceptor phototropin on excess absorbed energy dissipation (NPQ) and also demonstrated that besides light, downstream metabolism can also affect this acclimation process.Overall this Ph.D work reveals the existence and integration of different signal pathways in the regulation of photoprotective responses by microalgae living in the ocean and in the land

With increasing global populations and demand for energy, greater strain is placed on the limited supply of fossil derived fuels, which in turn drives the need for development of alternative energy sources. The discovery of biophotolysis in Chlamydomonas reinhardtii and the development of a spectral-selective photosystem I activating/photosystem II deactivating light (PSI-light) method provides a promising platform for commercial hydrogen production systems. The PSI-light method allows electrons to pass through the photosynthetic electron transport chain while reducing radiation available for photosynthetic oxygen evolution that inactivates hydrogenase. Exploring the physiology of photohydrogen production using the PSI-light method can provide insight on how to optimize conditions for maximum hydrogen production. Through the use of photosynthetic mutant strains of C. reinhardtii, it was possible to suppress photosynthetic oxygen evolution further than using photosystem I light alone to extend photohydrogen production longevity and total yield. A preliminary investigation of an iterating light treatment revealed that longevity and yield could be increased further by providing a period of darkness to allow cells to consume evolved oxygen and resynthesize hydrogenase. Work with these mutants provided understanding that a balance of radiation was required to provide electrons to hydrogenase while limiting oxygen evolution, and that when no light was provided, fermentation of stored starch was the major contributor of electrons to hydrogen production. To determine the role of starch during hydrogen production, wild type cells were exposed to different media and light treatments and monitored for starch consumption and hydrogen production. The results indicated that starch was required for hydrogen production in the dark, but for photohydrogen production, starch likely played a minor role in contributing electrons to hydrogenase. The experiments also showed the importance of acetate in the medium during the hydrogen production phase to allow any significant photohydrogen production. The role of acetate was further investigated as a growth medium constituent that stimulates metabolic activity while reducing photosynthetic oxygen evolution when added to cells grown auto- or mixotrophically. By exposing cells to CO₂ during growth, photohydrogen production was significantly increased over cells grown only in the presence of acetate.

Includes bibliographical references (leaves 235-258). Estimates the primary production and investigates the photosynthetic performance of temperate turfs at West Island, off the coast of South Australia. These communities play a fundamental role in reef ecology, being the main source of food for grazers, both fishes and invertebrates. Turfs also have an important function in benthic algal community dynamics, being the first colonizers on disturbed and bare substratum.

A prevailing view exists in the literature which suggests that macroalgae growing in slow-flow environments (<4 cm s⁻�) are less productive because of "mass-transfer" limitation compared to fast-flow environments. Macroalgae in slow-flow environments are thought to have thicker diffusion boundary-layers which limit the flux of essential molecules to and from the algal thallus. However nutrient demand of a macroalga can also influence nutrient flux. The main objective of this research was to determine the relative importance of physical (mainstream velocity) and physiological (nutrient demand) factors influencing the growth rate of Adamsiella chauvinii, a small (<20 cm) red algal species, that grows within the benthic boundary-layer in a soft sediment habitat. To establish the influence of water velocity, the growth rate of A. chauvinii was measured in situ each month (March 2003 to March 2004) at three sites with varying degrees of water velocity (slow, intermediate and fast) at which all other environmental parameters (photon flux density, seawater temperature and nutrients) were similar. To determine the metabolic demand and nutrient uptake rate of A. chauvinii, the internal nutrient status (C:N, soluble tissue nitrate, ammonium and phosphate), uptake kinetics (V[max] and K[s]) and nutrient uptake rate at a range of mainstream velocities were also determined on a seasonal basis. The hydrodynamic environment around A. chauvinii canopies was characterised in situ and compared with controlled laboratory experiments. Growth rates of Adamsiella chauvinii thalli at the slow-flow site were significantly lower in winter (June) to summer (February) than the intermediate- and fast-flow sites, while in autumn growth rates were similar between sites. However, A. chauvinii at the slow-flow site had similar or higher tissue N content compared to thalli at the other two sites during winter, spring and summer suggesting that growth rates of A. chauvinii were not mass-transfer limited. Nitrogen uptake rates of A. chauvinii were similar between sites in summer and winter, however uptake rates were lower in summer compared to winter even though thalli were nitrogen limited in summer. Water velocity had no effect on nitrate uptake in either summer or winter and uptake of ammonium increased with increasing water velocity during summer only. Two hydrodynamically different environments were distinguished over a canopy of A. chauvinii, with both the laboratory and field velocity profiles in good agreement with each other. In the top half of the canopy, the Turbulent Kinetic Energy (TKE) and Reynolds stresses were greatest while in the bottom half of the canopy flow rates were less than 90 % of mainstream velocity (< 1 cm s⁻�). When considered together, the influence of water velocity on the growth rates of A. chauvinii was not completely clear. Results suggest that mainstream velocity had little influence on nutrient availability to A. chauvinii because of the unique hydrodynamic environment created by the canopy. Nutrients, especially ammonium and phosphate, derived from the sediment and invertebrates, may provide enough nitrogen and phosphate to saturate the metabolic demand of Adamsiella chauvinii, consequently, A. chauvinii is well adapted to this soft-sediment environment.

Microalgae are a rich, largely untapped source of lipids. Algae are underutilized, in part, because lipid formation generally is stimulated by stress, such as nutrient deficiency. Nutrient deficiencies reduce growth, resulting in a tradeoff between elevated cellular lipids and abundant cell division. This tradeoff is not well understood. We also have a poor understanding of the physiological drivers for this lipid formation. Here we report on three sets of research: 1) Assessment of species differences in growth and lipid content tradeoffs with high and low level nitrogen deficiency; 2) Investigation of physiological drivers of lipid formation, by mass balance accounting of cellular nitrogen with progressing deficiency; 3) Examination of the effects of sodium chloride and silicon on lipid production in a marine diatom. 1) Nitrogen deficiency typically had disproportionate effects on growth and lipid content, with profound differences among species. Optimally balancing the tradeoff required a wide range in the rate of nitrogen supply to species. Some species grew first and then accumulated lipids, while other species grew and accumulated lipids concurrently--a characteristic that increased lipid productivity. High lipid content generally resulted from a response to minimal stress. 2) Commonalities among species in cellular nitrogen at the initiation of lipid accumulation provided insight into the physiological drivers for lipid accumulation in nitrogen deficient algae. Total nitrogen uptake and retention differed widely among species, but the ratio of minimum retained nitrogen to nitrogen at the initiation of lipid accumulation was consistent among species at 0.5 ± 0.04. This suggests that lipid accumulation was signaled by a common magnitude of nitrogen deficiency. Among the cellular pools of nitrogen at the initiation of lipid accumulation, the concentration of RNA and the protein to RNA ratio were most similar among species with averages of 3.2 ± 0.26 g L-1 (8.2% variation) and 16 ± 1.5 (9.2% variation), respectively. This implicates critical levels of these parameters as potential signals initiating the accumulation of lipids. 3) In a marine diatom, low levels of either sodium chloride or silicon resulted in at least 50% increases in lipid content. The synergy of simultaneous, moderate sodium chloride and silicon stress resulted in lipid content up to 73%. There was a strong sodium chloride/silicon interaction in total and ash-free dry mass densities that arose because low sodium chloride was inhibitory to growth, but the inhibition was overcome with excessive silicon supply. This suggests that low sodium chloride may have affected metabolism of silicon.

Several species of Bryopsidales (Chlorophyta) are known for displaying functional absence of the xanthophyll cycle, a common photoprotection mechanism responsible for qE component of NPQ. To cope with the extreme variability of their natural environment, these algae must be able to avoid photodamage. Previous works reported significant accumulation of all-trans-neoxanthin and violaxanthin under high light acclimation in different Bryopsidales, and speculated that these xanthophylls might control the amount of energy that reaches the photosystems, causing photoprotection. In this work, we investigated photoacclimation and photoprotection strategies in two species of Bryopsidales (Codium tomentosum and Bryopsis plumosa). We first characterised the acclimation state of algae exposed for 7 days to low light or high light (respectively 20 and 1000 μmol photons m2 s−1) in terms of pigment content (HPLC) and chlorophyll a variable fluorescence (PAM). We confirmed that high light triggers significant alteration of pigment content with accumulation of trans-Neoxanthin and Violaxanthin, and for the first time we characterised thoroughly how the pigment pool is altered during acclimation. We also confirmed that no evidence of any xanthophyll cycle is present in high light acclimated cultures. On a second note we tried to answer another major question: are trans-neoxanthin and violaxanthin photoprotective? Using a novel chlorophyll a variable fluorescence approach (pNPQ assessment) and performing quantification of PSII repair capacity (via lincomycin treatment) we were not able to give a clear answer to this question. Nonetheless, we concluded that despite trans-Neoxanthin and Violaxanthin might contribute to photoprotection, this process in Bryopsidales algae is likely given by the coordination between different mechanisms that deserve to be further investigated, including chloroplast movement, PSII repair/modulation, state transitions, and PSI cyclic electron transport.

Orientador: Orlando Necchi Júnior<br>Banca: Antonio Fernando Monteiro Camargo<br>Banca: Ricardo Moreira Chaloub<br>Resumo: Estudos sobre efeitos da manipulação de nutrientes inorgânicos (especialmente nitrogênio e fósforo) em macroalgas continentais são escassos, especialmente com espécies de regiões tropicais. Foram analisadas as respostas fisiológicas (crescimento, fotossíntese e conteúdo de pigmentos) ao empobrecimento de nitrogênio e fósforo inorgânicos (nas formas de nitrato e fosfato, respectivamente) em condições de cultura de duas populações de Compsopogon coeruleus (denominadas isolados 79 e 58) e em duas populações de Batrachospermum delicatulum (isolados 43 e 111), provenientes de ambientes com distintos níveis de saprobidade e trofia. O objetivo central foi avaliar as respostas das espécies (diminuição ou aumento na resposta fisiológica) conforme decréscimo nas concentrações de nitrogênio e fósforo inorgânicos. Três diluições na concentração original de nitrogênio e fósforo do Meio Básico de Bold foram empregadas: Bold (Controle), -10xN (0,21mM), -25xN(0,09mM), -50xN(0,04mM), - 10xP(0,11mM), -25xP(0,04mM) e -50xP(0,02mM). Nos tratamentos com nitrogênio, o isolado 79 de C. coeruleus respondeu com diminuição mais acentuada da performance geral em relação ao isolado 58, isto é, menores valores de rendimento quântico efetivo (RQE) e das concentrações das ficobilinas em todas as diluições de nitrogênio. A degradação de ficobilinas é uma resposta típica e amplamente descrita de Rhodophyta sob escassez de nitrogênio. O isolado 43 de B. delicatulum reduziu a síntese de clorofila com o decréscimo de nitrogênio no meio, enquanto que no isolado 111 o conteúdo de ficobilinas aumentou, sugerindo que estas populações possuem estratégias distintas de respostas da concentração de pigmentos em relação ao nitrogênio.<br>Abstract: Studies on the effects of inorganic nutrients manipulation (specially nitrogen and phosphorus) in freshwater macroalgae are scarce. Physiological responses (growth, photosynthesis and pigment contents) to nitrogen and phosphorus impoverishment (nitrate and phosphate, respectively) were analyzed under culture conditions in two populations of Compsopogon coeruleus (designated isolates 79 and 58), and of Batrachospermum delicatulum (isolates 43 and 111), coming from environments with distinct levels of saprobity and trophy. The essential aim was to evaluate the species' responses (decrease or increase in physiological performance) to decrease in inorganic nitrogen and phosphorus concentrations. Three dilutions in original concentration of nitrogen and phosphorus of Bold Basic Medium were tested: Bold (Control), -10xN (0,21mM), -25xN(0,09mM), -50xN(0,04mM), -10xP(0,11mM), -25xP(0,04mM) e - 50xP(0,02mM). For the nitrogen treatments, the isolate 79 of C. coeruleus responded with a more pronounced decrease in general performance in comparison to isolate 58, that is, lower values of effective quantum yield (EQY) and of phycobiliprotein concentrations in all nitrogen dilutions. Phycobiliprotein degradation is a typical and widely reported response of red algae under nitrogen scarcity. Isolate 43 of B. delicatulum reduced the chlorophyll synthesis according to nitrogen decrease in the medium, whereas in isolate 111 the phycobiliprotein content increased, suggesting that these populations have distinct strategies of pigment concentration responses to nitrogen impoverishment. For the phosphorus experiments, the isolate 58 responded with a more pronounced decrease in general performance in comparison to isolate 79: lower values of maximum photosynthetic rate (Pmáx) and photosynthetic efficiency (α), besides lower phycobiliprotein concentrations in all phosphorus dilutions.<br>Mestre

The in origin intertidal marine brown alga Fucus vesiculosus L. grow permanently sublittoral in the brackish Bothnian Sea, side by side with the recently discovered F. radicans L. Bergström et L. Kautsky. Environmental conditions like salinity, light and temperature are clearly different between F. vesiculosus growth sites in the Bothnian Sea (4-5 practical salinity units, psu; part of the Baltic Sea) and the tidal Norwegian Sea (34-35 psu; part of the Atlantic Ocean). The general aims of this thesis were to compare physiological aspects between the marine ecotype and the brackish ecotype of F. vesiculosus as well as between the two Bothnian Sea species F. vesiculosus and F. radicans. The result in the study indicates a higher number of water soluble organic compounds in the marine ecotype of F. vesiculosus compared to the brackish ecotype. These compounds are suggested to be compatible solutes and be due to an intertidal and sublittoral adaptation, respectively; where the intertidal ecotype needs the compounds as a protection from oxygen radicals produced during high irradiation at low tide. The sublittoral ecotype might have lost the ability to synthesize these compound/compounds due to its habitat adaptation. The mannitol content is also higher in the marine ecotype compared to the brackish ecotype of F. vesiculosus and this is suggested to be due to both higher level of irradiance and higher salinity at the growth site. 77 K fluorescence emission spectra and immunoblotting of D1 and PsaA proteins indicate that both ecotypes of F. vesiculosus as well as F. radicans have an uneven ratio of photosystem II/photosystem I (PSII/PSI) with an overweight of PSI. The fluorescence emission spectrum of the Bothnian Sea ecotype of F. vesiculosus however, indicates a larger light-harvesting antenna of PSII compared to the marine ecotype of F. vesiculosus and F. radicans. Distinct differences in 77 K fluorescence emission spectra between the Bothnian Sea ecotype of F. vesiculosus and F. radicans confirm that this is a reliable method to use to separate these species. The marine ecotype of F. vesiculosus has a higher photosynthetic maximum (Pmax) compared to the brackish ecotype of F. vesiculosus and F. radicans whereas both the brackish species have similar Pmax. A reason for higher Pmax in the marine ecotype of F. vesiculosus compared to F. radicans is the greater relative amount of ribulose-1.5-bisphosphate carboxylase/oxygenase (Rubisco). The reason for higher Pmax in marine ecotype of F. vesiculosus compare to the brackish ecotype however is not due to the relative amount of Rubisco and further studies of the rate of CO2 fixation by Rubisco is recommended. Treatments of the brackish ecotype of F. vesiculosus in higher salinity than the Bothnian Sea natural water indicate that the most favourable salinity for high Pmax is 10 psu, followed by 20 psu. One part of the explanation to a high Pmax in 10 psu is a greater relative amount of PsaA protein in algae treated in 10 psu. The reason for greater amount of PsaA might be that the algae need to produce more ATP, and are able to have a higher flow of cyclic electron transport around PSI to serve a higher rate of CO2 fixation by Rubisco. However, studies of the rate of CO2 fixation by Rubisco in algae treated in similar salinities as in present study are recommended to confirm this theory.<br>Fucus vesiculosus L. (Blåstång) är en brunalg som i huvudsak växer i tidvattenzonen i marint vatten men arten klarar också att växa konstant under ytan i det bräckta Bottenhavet. Norska havet och den del av Bottenhavet, där algerna är insamlade i denna studie, har salthalterna 34-35 psu (praktisk salthaltsenhet) respektive 4-5 psu. F. radicans L. Bergström et L. Kautsky (Smaltång) är en nyligen upptäckt art (2005) som har utvecklats i Bottenhavet. F. radicans och Bottenhavets ekotyp av F. vesiculosus växer sida vid sida och har tidigare ansetts vara samma art. Sett till hela Östersjön, så ändras ytans salthalt från 25 till 1-2 psu mellan Östersjöns gräns mot Kattegatt och norra Bottenviken. Den låga salthalten i Östersjön beror på det höga flödet av sötvatten från älvarna och på ett litet inflödet av saltvatten i inloppet vid Kattegatt. Salthaltsgradienten är korrelerad med antalet arter som minskar med minskad salthalt. Östersjön är ett artfattigt hav och de arter som finns är till stor del en blandning av söt- och saltvattenarter. Det finns bara ett fåtal arter som är helt anpassade till bräckt vatten och F. radicans är en av dem. Exempel på miljöskillnader för F. vesiculosus i Norska havet och i Bottenhavet är salthalten, tidvattnet, ljuset och temperaturen. Tidvattnet i Norska havet gör att algerna växlar mellan att vara i vattnet och på land, vilket utsätter algerna för stora ljusskillnader, snabba och stora temperaturväxlingar samt även torka. De alger som växer i Bottenhavet har däremot en jämnare och lägre temperatur, istäcke på vintern och mindre tillgång på ljus eftersom de alltid lever under vattenytan. Skillnaderna i miljön mellan växtplatserna leder till skillnader i fysiologiska anpassningar. Anledningen till att F. vesiculosus och F. radicans valdes som studieobjekt i denna avhandling är att de är viktiga nyckelarter i Bottenhavet. F. vesiculosus och F. radicans är de enda större bältesbildande alger som finns i det artfattiga ekosystemet och de används därför flitigt som mat, gömställe, parningsplats och barnkammare för t.ex. fisk. Att de är nyckelarter gör det angeläget att försöka förstå hur algerna är anpassade och hur de reagerar på miljöförändringar för att få veta hur de kan skyddas och bevaras. F. radicans inkluderades även för att se hur en naturlig art i Bottenhavet är anpassad i jämförelse med den invandrade F. vesiculosus. Marin F. vesiculosus inkluderades för att vara en artreferens från artens naturliga växtplats. Studien visar att det finns fler vattenlösliga organiska substanser (finns vissa organiska substanser som har en proteinskyddande funktion) i den marina ekotypen av of F. vesiculosus än i Bottenhavets ekotyp. Anledningen till detta föreslås vara en anpassning till att växa i tidvattenzonen. Vid lågvatten utsätts F. vesiculosus från Norska havet för starkt ljus, uttorkning, och snabba temperatur- växlingar vilket gör att den kan behöva dessa organiska substanser som skydd mot fria syreradikaler som bildas under lågvattenexponeringarna. F. vesiculosus från Bottenhavet har troligen mist förmågan att syntetisera dessa substanser på grund av anpassning till att hela tiden växa under ytan. Mängden mannitol (socker) är högre i den marina ekotypen av of F. vesiculosus än i Bottenhavets ekotyp. Detta föreslås bero på högre fotosyntetiskt maximum i F. vesiculosus från Norska havet jämfört med ekotypen från Bottenhavet. Skillnaden i fotssyntetiskt maximum är bland annat kopplat till ljus- och salthaltskillnaden på algernas växtplatser. Denna teori styrks av att både fotosyntesen och halten av mannitol ökar i Bottenhavets ekotyp när den behandlas i högre salthalt. Studien visar även att båda ekotyperna av F. vesiculosus samt F. radicans har ett ojämnt förhållande mellan fotosystem II och I (PSII och PSI) med en dominans av PSI. Denna slutsats är baserad på fluorescens emissions mätningar vid 77 K (-196 °C) och mätning av den relativa mängden D1 protein (motsvarar PSII) och PsaA protein (motsvarar PSI). F. vesiculosus från Bottenhavet visar ett emission spektrum som pekar mot en jämnare fördelning av PSII och PSI jämfört med den marina ekotypen och F. radicans. Detta stämmer dock inte med förhållandet mellan D1/PsaA som indikerar att alla tre har mer PSI än PSII. Förklaringen till avvikelsen mellan metoderna antas vara att F. vesiculosus från Bottenhavet har större ljus-infångande antennpigment än marin F. vesiculosus och F. radicans. De tydliga skillnaderna i 77 K fluorescens emission spektra mellan Bottenhavets F. vesiculosus och F. radicans visar att denna metod kan användas som säker artidentifiering. Den marina ekotypen av F. vesiculosus har högre fotosyntetiskt maximum än de båda arterna från Bottenhavet. Mätningar av den relativa mängden av enzymet Rubisco, viktigt för upptaget av koldioxid hos växter och alger, visar att mängden enzym är en sannolik förklaring till skillnaden i fotosyntetiskt maximum mellan den marina ekotypen av F. vesiculosus och F. radicans och detta är troligen en normal artskillnad. Mängden Rubisco kan dock inte förklara skillnaden i fotosyntetiskt maximum mellan de båda ekotyperna av F. vesiculosus. För att undersöka vad skillnaden mellan dessa två beror på så föreslås istället mätningar av Rubisco’s koldioxidfixeringshastighet. Det är en ökning av fotosyntetiskt maximum i Bottenhavets ekotyp av F. vesiculosus när den behandlas i högre salthalt (10, 20 och 35 psu) och det högsta fotosyntetiska maximumet uppmättes i alger som behandlats i 10 psu. Denna ökning beror inte på ökning i den relativa mängden av Rubisco. Ökningen i fotosyntesen speglas dock av en ökning av den relativa mängden PsaA. Detta antas bero på att det behövs mer energi i form av ATP och att en ökning av detta kan ske på grund av att mer PsaA kan driva den cykliska elektrontransporten i fotosyntesreaktionen. Ökat behov av ATP antas bero på en ökning av Rubisco aktiviteten men mätning av aktiviteten krävs för att bekräfta detta.

Le suivi de la qualité de l'eau a été d'une grande importance depuis ces dernières décennies afin de trouver des solutions de contrôler la contamination de l'eau, induite en grande partie par les activités agricoles et industrielles. Bien que les méthodes conventionnelles, comme la chromatographie, sont des outils très précis et sensibles, un intérêt grandissant a été placé sur des techniques prometteuses qui peuvent être utilisées sur site, sont bas coût, et offrent la possibilité d'effectuer des analyses rapides. Le travail présenté ici est dédié au développement de composant Laboratoire sur Puce pour l'analyse de la toxicité de l'eau. Il consiste en un système portable pour la détection sur site et offre la possibilité d'une double détection complémentaire : optique et électrochimique. Comme la partie dédiée au capteur électrochimique a préalablement été validée, cette étude est focalisée sur l'implémentation d'un biocapteur électrochimique basé sur l'utilisation d'une algue, pour la détection de polluants dans l'eau. Le principe basique de détection consiste au suivi de changements de l'activité métabolique d'algues induits par la présence d'herbicides. La réponse de l'algue est différente pour chaque concentration d'herbicide dans un échantillon examiné. Deux herbicides sélectionnés affectent l'activité photosynthétique de l'algue et par conséquent, induisent des modifications dans la quantité des espèces électroactives produites par l'algue : O2, H2O2 et H3O+/OH-. Avant le développement du composant final type Laboratoire sur Puce, les principes de détection aussi bien que les matériaux d'électrode qui vont être intégrés, ont été validés en utilisant un type de composant plus simple, qui a été réalisé grâce aux technologies de fabrication silicium et qui a été caractérisé par des procédures plus simples. Une puce sur silicium contenant un microsystème électrochimique intégrant trois électrodes a été mis en place. Une fois validés, les matériaux de détection et les configurations choisis précédemment ont été utilisés pour la fabrication des composants Laboratoire sur Puce. Les composants Laboratoire sur Puce ont été ensuite utilisés pour des tests biologiques afin de détecter les herbicides d'intérêt. Une attention spéciale a été placée sur le suivi de O2 comme indicateur de la présence d'herbicide, étant donné que cet élément est le plus représentatif de modifications de l'activité métabolique. Un effet d'inhibition sur la photosynthèse, dépendant de la concentration de l'herbicide a été démontré. La détection de l'herbicide a été réalisée avec une grande sensibilité et sur une gamme couvrant la limite de concentration maximale acceptable imposé par le gouvernement canadien<br>Water quality assessment has attracted wide attention during the last decades in order to find ways to control contamination of water bodies induced, in a big part, by agricultural and industrial activities. Although conventional techniques, such as chromatography are highly accurate and sensitive tools, increasing interest has been placed lately to powerful alternative techniques that can be used on field, are cost-effective and offer the possibility of conducting rapid analysis. The present work was therefore dedicated to the development of a lab-on-chip device for water toxicity analysis. It consists in a portable system for on-site detection and aims at offering the possibility of conducting double complementary detection: optical and electrochemical. Since the optical sensor is already validated, this study focused on the implementation of the algal-based, electrochemical biosensor for detection water contaminants. The basic detection principle consists in monitoring disturbances in metabolic activities of algae induced by the presence of the herbicides. Algal response is different for each herbicide concentration in the examined sample. The two selected herbicides affect algal photosynthetic activity and consequently induce modifications in the quantity of electroactive species, O2, H2O2 and H3O+/OH- ions related to pH, produced by algae. Prior to the development of the final lab-on-chip device, the detection principle as well as the electrode materials that were going to be integrated were validated using a simpler device that was implemented using a silicon-based fabrication technology and was characterized using simpler procedures. A silicon chip containing the integrated three-electrode electrochemical microsystem was fabricated. The performance of the microsystem was evaluated through electrochemical characterization and calibration was performed. Once validated, the aforementioned materials and configurations were used for the fabrication of the lab-on-chip devices. The lab-on-chip devices were further used in bioassays to detect the herbicides of interest. Special emphasis was placed on O2 monitoring as indicator of the presence of herbicide, as it is the element the most representative of variations in metabolic activities. A concentration-dependent inhibition effect of the herbicide on photosynthesis was demonstrated. Herbicide detection was achieved with a greater sensitivity and a range covering the limit of maximum acceptable concentration imposed by Canadian government

Made available in DSpace on 2014-06-11T19:23:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-25Bitstream added on 2014-06-13T19:49:37Z : No. of bitstreams: 1 bautista_ain_me_rcla.pdf: 374564 bytes, checksum: 1e7cf9c5da15f9f6f26f1fe790157e7c (MD5)<br>Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)<br>Estudos sobre efeitos da manipulação de nutrientes inorgânicos (especialmente nitrogênio e fósforo) em macroalgas continentais são escassos, especialmente com espécies de regiões tropicais. Foram analisadas as respostas fisiológicas (crescimento, fotossíntese e conteúdo de pigmentos) ao empobrecimento de nitrogênio e fósforo inorgânicos (nas formas de nitrato e fosfato, respectivamente) em condições de cultura de duas populações de Compsopogon coeruleus (denominadas isolados 79 e 58) e em duas populações de Batrachospermum delicatulum (isolados 43 e 111), provenientes de ambientes com distintos níveis de saprobidade e trofia. O objetivo central foi avaliar as respostas das espécies (diminuição ou aumento na resposta fisiológica) conforme decréscimo nas concentrações de nitrogênio e fósforo inorgânicos. Três diluições na concentração original de nitrogênio e fósforo do Meio Básico de Bold foram empregadas: Bold (Controle), -10xN (0,21mM), -25xN(0,09mM), -50xN(0,04mM), - 10xP(0,11mM), -25xP(0,04mM) e -50xP(0,02mM). Nos tratamentos com nitrogênio, o isolado 79 de C. coeruleus respondeu com diminuição mais acentuada da performance geral em relação ao isolado 58, isto é, menores valores de rendimento quântico efetivo (RQE) e das concentrações das ficobilinas em todas as diluições de nitrogênio. A degradação de ficobilinas é uma resposta típica e amplamente descrita de Rhodophyta sob escassez de nitrogênio. O isolado 43 de B. delicatulum reduziu a síntese de clorofila com o decréscimo de nitrogênio no meio, enquanto que no isolado 111 o conteúdo de ficobilinas aumentou, sugerindo que estas populações possuem estratégias distintas de respostas da concentração de pigmentos em relação ao nitrogênio.<br>Studies on the effects of inorganic nutrients manipulation (specially nitrogen and phosphorus) in freshwater macroalgae are scarce. Physiological responses (growth, photosynthesis and pigment contents) to nitrogen and phosphorus impoverishment (nitrate and phosphate, respectively) were analyzed under culture conditions in two populations of Compsopogon coeruleus (designated isolates 79 and 58), and of Batrachospermum delicatulum (isolates 43 and 111), coming from environments with distinct levels of saprobity and trophy. The essential aim was to evaluate the species’ responses (decrease or increase in physiological performance) to decrease in inorganic nitrogen and phosphorus concentrations. Three dilutions in original concentration of nitrogen and phosphorus of Bold Basic Medium were tested: Bold (Control), -10xN (0,21mM), -25xN(0,09mM), -50xN(0,04mM), -10xP(0,11mM), -25xP(0,04mM) e - 50xP(0,02mM). For the nitrogen treatments, the isolate 79 of C. coeruleus responded with a more pronounced decrease in general performance in comparison to isolate 58, that is, lower values of effective quantum yield (EQY) and of phycobiliprotein concentrations in all nitrogen dilutions. Phycobiliprotein degradation is a typical and widely reported response of red algae under nitrogen scarcity. Isolate 43 of B. delicatulum reduced the chlorophyll synthesis according to nitrogen decrease in the medium, whereas in isolate 111 the phycobiliprotein content increased, suggesting that these populations have distinct strategies of pigment concentration responses to nitrogen impoverishment. For the phosphorus experiments, the isolate 58 responded with a more pronounced decrease in general performance in comparison to isolate 79: lower values of maximum photosynthetic rate (Pmáx) and photosynthetic efficiency (α), besides lower phycobiliprotein concentrations in all phosphorus dilutions.

Les populations de bivalves exploités subissent régulièrement des épizooties qui affaiblissent voire déciment les stocks, et qui peuvent avoir des conséquences majeures pour l’aquaculture. Ces maladies, dues à des virus, bactéries, ou parasites, se développent particulièrement au printemps et en été. Ces périodes de l’année offrent également des conditions propices aux efflorescences de micro-algues toxiques, dont des dinoflagellés du genre Alexandrium. Ainsi, le risque de co-occurrence d’efflorescences d’Alexandrium sp. et de maladies infectieuses chez les bivalves est élevé. Or, ces micro-algues synthétisent et excrètent des neurotoxines et des composés cytotoxiques responsables d’altérations physiologiques chez les bivalves. L’objectif de cette thèse est d’évaluer les effets combinés d’une exposition à Alexandrium sp. et d’une infection par des agents pathogènes sur la physiologie des bivalves, à travers l’étude de différentes interactions tripartites bivalve – pathogène – Alexandrium sp. Les résultats de ce travail indiquent que différents profils de réponse existent en fonction des espèces impliquées dans ces interactions. Ainsi, une exposition à Alexandrium sp. peut augmenter le taux d’infection par des agents pathogènes chez des bivalves ou au contraire le diminuer. Les réponses hémocytaires associées peuvent traduire l’implication des défenses immunitaires dans ces modulations hôte-pathogène. De plus, l’exposition à des agents pathogènes peut interférer avec le processus d’accumulation de toxines algales dans les tissus des bivalves, illustrant la complexité de ces interactions. Ces résultats, associés à l’observation de lésions tissulaires chez les bivalves peuvent traduire l’altération des activités de nutrition (filtration, digestion…). Ce travail de thèse apporte une meilleure compréhension de l’implication des efflorescences toxiques dans le développement des maladies touchant les bivalves d’intérêt commercial, mais également de l’implication de l’environnement biotique des bivalves sur l’accumulation de phycotoxines réglementées<br>Bivalve populations undergo regular epidemics that weaken or decimate exploited stocks and thus limit aquaculture. These diseases are caused mainly by viruses, bacteria or parasites, and occur primarily during spring and summer. This period of the year also provides favorable conditions for toxic dinoflagellate blooms, including species of the genus Alexandrium. Thus, the risk of Alexandrium sp. blooms and infectious diseases co-occurring in bivalves is high. However, these micro-algae synthesize and excrete toxins and cytotoxic compounds responsible for physiological changes in bivalves and could lead to an immuno-compromised status.The objective of this thesis is to evaluate the combined effects on bivalve physiology of exposure to the toxic dinoflagellate, Alexandrium sp., and infection by pathogens, through the study of different bivalve - pathogen - Alexandrium sp. tripartite interactions. The results of this work highlight the species-specific nature of these impacts.Thus, exposure to Alexandrium catenella reduces the herpesviruses infection in oyster Crassostrea gigas, whereas the dinoflagellate A. fundyense increases the susceptibility of C. virginica oyster to the parasite Perkinsus marinus, probably via immuno-suppression, as suggested by the partial inhibition of hemocyte responses. Additionally, the effect of a toxic algal bloom on oyster susceptibility to opportunistic diseases when exposed to a new microbial environment (simulating a transfer) was evaluated. Hemocyte responses to a changing microbial environment were suppressed by exposure to A. catenella, although no new bacterial infection was detected.Finally, exposure to pathogens or to a new microbial environment interferes with the processes by which oysters exposed to A. catenella accumulate algal toxins, illustrating the complexity of these interactions. These results provide a better understanding of the involvement of toxic algal blooms in the development of diseases affecting commercial bivalve species, but also of the involvement of the bivalve biotic environment in the accumulation of regulated toxins

Harmful algal blooms (HABs) are found worldwide, particularly in places where warm, well-lit, and stagnant waters are common. HABs can have negative effects on aquatic plants and wildlife due to the reduction in light availability associated with turbidity, decrease in O2 availability, and the production of secondary metabolites that can harm or even prove lethal. Aquatic ecosystems are regularly being affected by elevated salinity because of recent water management strategies, episodes of drought, and salt water intrusion. This research focused on how salinity levels ranging from 0-10ppt affected physiological attributes such as cellular growth and abundance, cell mortality, toxin release, and oxidative stress in the toxigenic cyanobacterium, Microcystis aeruginosa. It was determined that salinity treatments of 7ppt and above caused a decrease in both cellular growth and abundance, as well as an increase in toxin release due to cell mortality. M. aeruginosa was able to survive in salinities up to 7ppt. A pattern of caspase activity in response to elevated salinity was shown, but whether cellular mortality was due solely to programmed cell death (PCD) was not definitive. A strong antioxidant response, measured through catalase activity, was noted when salinity was enhanced to 7ppt. Above this value, the damaging effects of salinity caused elevated levels of reactive oxygen species (ROS) production and cell death. It was determined that the maximum amount of hydrogen peroxide that M. aeruginosa could withstand without significant impact to growth and abundance was below 250µM. Salinities of 7ppt and above had a negative impact on the physiology of M. aeruginosa, leading to cell death and an increase in microcystin release into the environment. These two factors can lead to fish kills, poor drinking water, and other recreational and commercial problems for an aquatic ecosystem. By determining the precise salinity that HAB cellular mortality is imminent, predictive models can be employed to predict the impacts of salt intrusion and groundwater management.

Orientadores: Maria Regina Wolf Maciel, Reinaldo Gaspar Bastos<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química<br>Made available in DSpace on 2018-08-22T08:48:28Z (GMT). No. of bitstreams: 1 Klein_BrunoColling_M.pdf: 2562214 bytes, checksum: f891de86d253786cf5d2101fec1f3eba (MD5) Previous issue date: 2013<br>Resumo: A busca por uma maior sustentabilidade tem levado a uma mudança em direção à utilização de fontes renováveis para geração de energia em detrimento do uso de combustíveis fósseis, visando a uma modificação na matriz energética global. A utilização da biomassa de microalgas para produção de biocombustíveis vem sendo vista como uma alternativa promissora, uma vez que o seu cultivo proporciona produtividades em carboidratos e lipídios superiores às matérias-primas vegetais convencionalmente utilizadas na obtenção de etanol e biodiesel. Neste contexto, o objetivo da presente dissertação de mestrado foi avaliar a produção de biomassa da microalga clorofícea Chlorella vulgaris em fotobiorreator de placa plana em diferentes condições de fluxo luminoso, concentração de CO2 na alimentação gasosa e concentração de NaNO3 no meio de cultivo, visando o acúmulo de carboidratos para obtenção de bioetanol de terceira geração. As influências das variáveis nutricionais e de processo sobre a eficiência fotossintética das microalgas também foram estimadas para determinação do estado fisiológico das culturas. A produtividade média de biomassa e a concentração máxima final das microalgas foram significativamente afetadas pela incidência de radiação luminosa e pela suplementação de CO2 gasoso, obtendo-se maiores produtividades de carboidratos em cultivos com alto fluxo luminoso e concentrações de CO2 intermediárias (7,5%). Também foi observado o efeito positivo do aumento do fotoperíodo sobre o crescimento das microalgas. Através de hidrólise ácida foi possível atingir concentrações de até 2 g L-1 de açúcares fermentescíveis no hidrolisado a partir de biomassa de microalgas cultivadas em meio com baixo teor de nitrogênio. A fermentação etanólica foi então conduzida com a levedura Dekkerabruxellensis capaz de converter diferentes hexoses e pentoses em bioetanol, dada a presença de ambos os tipos de açúcares no hidrolisado<br>Abstract: The search for industrial processes with higher sustainability has led to a change towards the utilization of renewable sources for energy generation in substitution of fossil fuels, aiming the modification of the global energy matrix. The utilization of microalgal biomass for the production of biofuels is viewed as a promising alternative, since its cultivation yields carbohydrate and lipid productivities superior to those of conventional sources used in the obtention of bioethanol and biodiesel. In this context, the goal of this master thesis was to evaluate the biomass production of the chlorophycean microalga Chlorella vulgaris in a flat plate photobioreactor under different conditions of light flux, CO2 concentration in the gas feed and NaNO3 concentration in the culture medium, aiming carbohydrate accumulation for the production of third generation bioethanol. The influences of both process and nutritional variables on the photosynthetic efficiency of microalgae were estimated for the determination of the physiological condition of the cultures. The mean biomass productivity and the maximum final microalgae concentration were significantly affected by the incidence of light radiation and by the supplementation of gaseous CO2, the highest carbohydrate productivities being obtained in cultivations with high light flux and intermediate CO2 concentrations (7,5%). It was also observed the positive effect of increasing the photoperiod over microalgae growth. Through acid hydrolysis, it was possible to attain fermentable sugar concentration of up to 2 g L-1 from biomass of microalgae cultivated in low-nitrogen medium. The ethanolic fermentation was then carried out with the Dekkerabruxellensis yeast, capable of converting different hexoses and pentoses into ethanol, due to the presence of both sugar types in the hydrolysate<br>Mestrado<br>Desenvolvimento de Processos Químicos<br>Mestre em Engenharia Química

Pour produire l’énergie nécessaire au métabolisme cellulaire, les eucaryotes photosynthétiques se servent de deux organites : le chloroplaste et la mitochondrie. Le premier est capable de convertir l’énergie lumineuse en énergie chimique et la deuxième est le lieu de la phosphorylation oxydative. Puisque les deux organites partagent la même fonction de production d’énergie, leurs activités doivent être régulées. Chez les plantes et les algues vertes ces deux « moteurs » cellulaires travaillent principalement de façon indépendante selon les conditions environnementales, comme la présence de sucre ou de lumière. Au contraire, chez les diatomées, l’échange direct de ATP et de NAPDH entre ces organites est essentiel pour assurer la survie de la cellule. Bien que cette interaction énergétique ait été caractérisée d’un point de vue physiologique, les acteurs moléculaires responsables de ce processus restent encore inconnus. Dans le cadre de mon projet de thèse, quatre protéines candidates ont été sélectionnées et étudiées pour déterminer leur implication dans les échanges énergétiques entre ces deux organites. Ces protéines sont des transporteurs localisés au niveau de l’enveloppe chloroplastique et de la membrane interne de la mitochondrie. Pour comprendre leur rôle physiologique, les souches mutantes correspondantes ont été générées. La capacité photosynthétique et la respiration cellulaire de ces mutants ont été évaluées par des approches de fluorescence, de spectroscopie, ou de mesure du taux d’évolution de l’oxygène. Ces mesures suggèrent que les transporteurs sélectionnés contrôlent en partie les mécanismes des échanges énergétiques entre le chloroplaste et la mitochondrie bien que d’autres protéines (non identifiées) semblent aussi impliquées dans cette régulation. Une caractérisation plus avancée de ces transporteurs pourrait permettre d’augmenter la production de biomasse des microalgues dans le cadre d’applications biotechnologiques, en favorisant l’utilisation simultanée de la respiration et de la photosynthèse (mixotrophie)<br>To produce the energy needed for cell metabolism, eukaryotic photosynthetic organisms rely on two organelles: the chloroplast and the mitochondrion. The former converting light energy into chemical energy, the latter performing cell respiration. Since both organelles have overlapping function, their activities need to be regulated. While in plants and green algae they seem to work overall independently according to environmental conditions, like light and sugar availability, in Diatoms the direct exchange of ATP and NADPH between these two organelles are essential for the cell’s survival. Although the physiology of this energetic crosstalk is well established, the molecular actors of this process are still unknown. During this PhD project, I have selected four candidate proteins, which turned out to play a role the organelles’ cross talk mechanisms. These are transporters predicted to be located within the chloroplast envelope and the inner membrane of the mitochondrion. To understand their physiological role, we compared the photosynthetic performances of the wildtype and the mutant strains with spectroscopic and fluorescence approaches, while the respiration was quantified measuring the oxygen evolution rates.The observed differences suggest that the selected transporters play a role the chloroplast-mitochondrion crosstalk and that other proteins might be involved in this regulative process.The further characterization of these transporters might also validate them as possible targets to improve algal biomass productivity for biotech, promoting the simultaneous use of respiration and photosynthesis (mixotrophy)

Les euglenes cultivees en presence du substrat glycolate utilisent le systeme chloroplastique pour synthetiser leurs reserves carbonees et s'apparentent aux euglenes photoautotrophes par leur photochimie du psii, leurs activites carboxylasiques et leur ultrastructure

2006/2007<br>RIASSUNTO Il recente Codice dell’Ambiente (D.lgs 152/2006) prevede il raggiungimento di ‘standard’ di qualità dei corpi idrici con particolare riferimento ai metalli pesanti (sostanze pericolose prioritarie, PP). Il passato piano di monitoraggio (2001-2004) del Ministero dell’Ambiente ha messo in luce il frequente superamento dei limiti di legge nelle stazioni del Golfo di Trieste. I metodi chimico/fisici utilizzati per determinare le concentrazioni di metalli in acqua o nei sedimenti non sono in grado di determinare l’effettiva biodisponibilità degli elementi, da cui dipende la reale tossicità, né un preciso rapporto causa/effetto. Tuttavia, è risaputo che anche basse concentrazioni possono innescare fenomeni di bioaccumulo e di biomagnificazione, alterando gli equilibri naturali soprattutto negli ambienti marini costieri. Tali processi interessano principalmente la componente macroalgale, molto importante dal punto di vista ecologico ed economico in quanto è alla base della catena trofica. Lo studio dell’impatto delle sostanze tossiche su questa matrice risulta, perciò, fondamentale per preservare da una parte la biodiversità degli ecosistemi dall’altra proporre sistemi alternativi/innovativi per il mantenimento della capacità auto-depurativa. Per le alghe e per le Cianoficee, in particolare, è stata dimostrata un’elevata capacità di difesa contro la tossicità di diversi metalli attraverso vari e complessi meccanismi, le cui modalità sono abbastanza conosciute. E’ oggi noto che le risposte difensive possono dipendere dalla natura del metallo, dalle dosi e dai tempi di esposizione, dai differenti stadi di crescita, e, non ultimo, dalla diversa ‘sensibilità’ delle alghe. Tuttavia, negli studi in campo ed in laboratorio, i diversi approcci sperimentali rendono la comparazione, sia nell’ambito della stessa specie che tra specie diverse, estremamente difficile e possono portare alla determinazione di dosi subletali (LC50) molto diverse. Da qui la necessità di un protocollo unificato che, a differenza delle micro-alghe, non è ancora standardizzato per le macro-alghe. Tale mancanza ha comportato, infatti, risultati molto diversi senza discriminazione delle risposte adattative da quelle difensive. Inoltre, raramente vengono analizzati l’eventuale ripristino della funzionalità cellulare durante il recupero in assenza del metallo ed i possibili effetti a livello ultrastrutturale. Scopo, percorso del progetto e metodi utilizzati Il progetto si basa principalmente sull’approfondimento delle conoscenze dei meccanismi di adattamento/tolleranza/difesa contro il Cadmio di micro- e macro-alghe marine a livello fisiologico, biochimico ed ultrastrutturale. L’obiettivo principale è rivolto alla possibilità di mettere a punto un protocollo standardizzato per le macroalghe in grado di discriminare specie ‘sensibili’, utilizzabili come ‘bioindicatori’ nei saggi ecotossicologici, da quelle utili come bioaccumulatori. Nel corso dei tre anni il materiale bibliografico, selezionato e continuamente aggiornato, è stato inserito in un archivio digitale con i dati relativi alle specie algali, ai metalli, alle dosi/tempi di esposizione, ai parametri saggiati con le rispettive metodiche e alle dosi soglia di tossicità. Una parte dei lavori può essere consultata a video (PDF) attraverso un collegamento ipertestuale (dati non presentati). L’attività di ricerca è stata suddivisa in tre parti: Parte I. Studio su micro-alghe. Sono stati approfonditi i meccanismi di adattamento/difesa ed i principali ‘siti bersaglio’ a livello fisiologico ed ultrastrutturale della Cyanoficea coloniale (Leptolyngbya sp.) mantenuta in coltura. Lo studio è mirato soprattutto alla valutazione del metodo fluorimetrico (PAM), di nuova acquisizione nel nostro laboratorio, scelto per la minor invasività rispetto alle altre analisi (ossimetria e pigmenti). Parte II. Studio su macro-alghe. La ricerca è stata condotta in tre fasi per individuare e scegliere specie sensibili/tolleranti, valutare la potenzialità delle specie considerabili buoni ‘bioaccumulatori’ e verificarne le potenzialità d’uso come ‘biofiltri riciclabili’, attraverso la stima del recupero funzionale in assenza del tossico. Parte III. Studio su macro-alghe in situ. E’ stato valutato il bioaccumulo dei popolamenti presenti nella Baia di Punta Olmi (Località ‘Boa’ – Muggia), area inquinata da metalli pesanti. Le specie macroalgali sono state prelevate in due in due siti costieri (Baia Punta Olmi – Muggia e Porto Franco Vecchio - Trieste). Dopo la stima delle curve di saturazione i campioni sono stati adattati in cella di coltura in condizioni controllate. Al mezzo di coltura sono state aggiunte dosi di cadmio [Cd(NO3)2] e le analisi, differenziate per le diverse fasi sperimentali, sono state effettuate dopo 1-2-7 giorni di ‘stress’ e dopo 7 giorni di recupero. Sono state stimate la fotosintesi (ossimetria e fluorimetria PAM), la respirazione, il contenuto di pigmenti liposolubili e condotte osservazioni al TEM. Su alcuni campioni è stata effettuata la microanalisi ai Raggi-X (SEM-EDS) variando la modalità di allestimento dei preparati. Conclusioni La stima delle condizioni di coltura ed il controllo incrociato dei risultati di questa ricerca forniscono le basi per interessanti sviluppi a livello interpretativo e sembrano essere un buon punto di partenza per valutare in modo più analitico le informazioni sulle possibili alterazioni del processo fotosintetico. La tecnica fluorimetrica offre, perciò, indubbi vantaggi in quanto non invasiva/distruttiva, ma soprattutto in relazione al numero dei parametri disponibili. Per l’applicazione corretta del metodo fluorimetrico è risultato fondamentale verificare i vantaggi ed i limiti dello strumento. L’analisi contemporanea di più parametri (ecologici, fisiologici, biochimici, ultrastrutturali), con dosi e tempi fissi, dipendendo da quest’ultimi l’ampiezza della risposta, è un approccio valido per individuare e descrivere in maniera più completa le risposte di adattamento/difesa delle alghe contro gli agenti tossici. Per quanto attiene ai tempi di esposizione, gli intervalli delle analisi (1S-2S-7S) sono risultati sufficienti per la valutazione delle risposte fisiologiche, biochimiche ed ultrastrutturali allo ‘stress’, mentre i 7 giorni di recupero in assenza del metallo (7R) sono risultati sufficienti per valutare le diverse capacità di ripresa funzionale delle specie. Le tecniche ultrastrutturali si confermano indispensabili per la localizzazione dei siti di sequestro ed accumulo del metallo, per lo studio delle modifiche cellulari, e per l’interpretazione dei risultati fisiologici. Diversamente, la microanalisi ai Raggi-X con il programma attualmente in dotazione (SEM-QUANT, ZAF), non è una tecnica valida per stimare il grado di bioaccumulo in popolamenti naturali, poiché le basse concentrazioni di metalli nei campioni biologici risultano spesso inferiori al limite di rilevabilità del sistema. Le diverse fasi sperimentali hanno permesso di identificare le specie più sensibili al cadmio, che potrebbero essere utilizzate in laboratorio come specie ‘test’ nei saggi ecotossicologici e in campo come ‘biosensori’. Quelle tolleranti/resistenti sarebbero, invece, utili come bioaccumulatori. In relazione ai tempi di esposizione, alle dosi applicate e sulla base dei risultati fisiologici ed ultrastrutturali mediante i quali sono stati messi in evidenza i principali siti di accumulo, sono state individuate specie con potenzialità diverse: a) P. pavonica, S. lomentaria, R. pseudopalmata, Z. typus e C.multifida come specie ‘sensibili’; b) R. thysanorhizans come specie ‘sensibile’ di ambiente naturale; c) H. musciformis, C. fragile, D. dichotoma come specie tolleranti/resistenti; d) F. virsoides e S. scoparium come specie tolleranti/resistenti con pronto recupero della piena funzionalità in assenza del tossico. Sono queste, quindi, le potenziali candidate da utilizzare come ‘biofiltri’ nel biorimedio. Al fine di poter utilizzare micro- e soprattutto macro-alghe (per l’abbondante biomassa), come biofiltri nel biorimedio, senza depauperamento delle risorse naturali, ed evitare nel contempo problemi di smaltimento del materiale contaminato, appare necessario non solo selezionare ‘buoni bioaccumulatori’ (specie tolleranti e/o resistenti), anche attraverso stime quantitative degli elementi per via chimica (spettrometria di massa), ma anche valutare le loro capacità di detossificarsi e di recuperare la piena funzionalità fisiologica in assenza del tossico. Solo in questo modo le alghe possono essere riutilizzate più volte per assorbire nuovo metallo.<br>XX Ciclo<br>1972

Les fortes intensités lumineuses, et notamment l’augmentation actuelle de l’intensité des radiations UV-B due à l’amincissement de la couche d’ozone, peuvent être des facteurs stressants majeurs pour la plupart des organismes phototrophes peuplant les écosystèmes terrestres et aquatiques. L’algue brune Pelvetia canaliculata et l’angiosperme des marais salés littoraux Salicornia ramosissima sont deux végétaux marins peuplant les hauts niveaux de l’estran, zone où ils sont soumis à de longues périodes d’exondation, exposés de manière directe au rayonnement solaire et susceptibles de posséder des mécanismes de photoprotection convergents efficaces. Dans cette étude, grâce à la mise au point d’un système de co-culture, leurs réponses métaboliques contre l’exposition â différentes niveaux de radiations UV-B ont été comparées en conditions contrôlées, notamment la synthèse de composés photoprotecteurs impliqués dans le maintien de la photosynthèse par la filtration des radiations UV et la défense contre le stress oxydatif. Les résultats montrent que les composés phénoliques semblent être la classe majoritaire de composés photoprotecteurs impliquée chez les deux espèces, avec un rôle écran préférentiel des flavonoïdes chez Salicornia ramosissima et une forte activité antioxydante des phlorotannins chez Pelvetia canaliculata. Cependant, les deux espèces pourraient moduler la composition de leur pool de composés phénoliques en jouant sur une de ces activités protectrices ou sur les deux, en fonction des conditions lumineuses. La fonction spécifique de filtration des radiations UV des composés photoprotecteurs a été mise en évidence après le développement d’une nouvelle méthode de mesure in vitro de l’activité écran d’extraits végétaux<br>High light irradiances, especially enhanced UV-B radiations due to present stratospheric ozone depletion, may be major stress factors for many phototrophic organisms in both aquatic and terrestrial ecosystems. The brown seaweed Pelvetia canaliculata and the coastal salt marsh angiosperm Salicornia ramosissima are two marine plants growing in the upper intertidal zone, where they are subjected to long emersion periods, to direct solar exposure and are likely to have convergent efficient photoprotective mechanisms. In this study, thanks to the development of a coculture system, their metabolic responses against various UV-B radiation levels have been compared in controlled conditions, especially the synthesis of photoprotective compounds implied in the photosynthesis maintaining by either a UV-absorbing function or a defence capacity against oxidative stress. Results show that phenols seem to be the main class of photoprotective compounds in both species, with a preferential sunscreen function of flavonoids in Salicornia ramosissima and a strong antioxidant activity of phlorotannins in Pelvetia canaliculata. However, both species could modulate the composition of their phenol pool through either 011e protective function or both depending on light conditions. The specific UV-absorbing function of these compounds has been evidenced thanks to the development of a new in vitro method for evaluating the sunscreen activity of plant extracts

Etude de la composition osidique d'alginate des genres pelvetia, fucus, arcophyllum, sargassum et laminaria. Preparation et caracterisation des alginates-lyases a partir de divers mollusques marins et d'une bacterie marine. Etude comparee de la regeneration de la paroi du protoplaste et de la mise en place normale de la paroi du zygote de fucus distichus par marquage avec des anticorps monoclonaux

Influence de differentes intensites d'eclairement sur la ramification du thalle, l'organisation et la differenciation du chloroplaste. Sur la croissance avec action du phytochisme, sur la photosynthese, le metabolisme du glycolate et glucides solubles et insolubles

Dans le cadre de la conservation du patrimoine, les micro-organismes se développant sous forme de biofilm sont souvent considérés comme des agents entraînant des problèmes esthétiques et de détérioration du support colonisé. L’objectif général de cette thèse est l’utilisation de la lumière UV-C comme méthode de traitement des biofilms. Dans le but de comprendre la diversité des biofilms, les communautés bactériennes, fongiques et des protistes autotrophes ont été étudiées à l’aide d’une technique de séquençage haut débit. Puis, afin de comprendre les effets des UV-C sur les micro-algales (Chlorella sp.), des essais en laboratoire ont permis de caractériser les réponses physiologiques algales (mort cellulaire, dégradation de la chlorophylle, état de photosystème II, etc.) suite à différents traitements aux UV-C. Tous les dommages observés au niveau cellulaire et moléculaire (dont la mort cellulaire et le « bleaching » de la chlorophylle) ont permis d’optimiser la méthode de traitement. Dans le but de s’assurer de l’innocuité du traitement aux UV-C sur le support des biofilms (peintures pariétales), des pigments et des liants, utilisés à la préhistoire, ont été fortement irradiés. Les résultats ont montré que les composés inorganiques sont insensibles aux UV-C au contraire des molécules organiques. Enfin, le traitement UV-C a démontré son efficacité dans une grotte touristique en permettant d’éradiquer tous les biofilms. De plus, aucune recolonisation n’a été observée deux ans après le traitement, démontrant que la lumière UV-C est le traitement le plus écologique à ce jour<br>Development of biofilm on monument heritage are often considered as agents leading to aesthetic issues and biodeterioration of their support. The aim of this study is the use of UV-C light as alternative treatment against Lampenflora. Bacterial, algal and fungal communities were studied using new generation sequencing approach. Following UV-C treatments, physiological responses of Chlorella sp. were studied in laboratory (cell death, chlorophyll degradation, photosystem II status, etc.). Damages at cellular and molecular levels (cell death, bleaching of the chlorophyll) were taken into consideration to optimize the UV-C treatment. Then, to ensure the safety of UV-C for biofilms support (parietal paintings), several pigments and binders, used by prehistoric human, were highly irradiated. The results showed that UV-C radiations were not deleterious for inorganic compounds, while a color change was observed on organic binders. Finally, UV-C treatment has proven its effectiveness in La Glacière show cave by eradicating all biofilms. In addition, no recolonization was observed two years after treatment, showing that this treatment, which is the most environmental friendly to date, is effective over time

The Huon Estuary is a micro-tidal estuary in south-east Tasmania that is an important area for salmon aquaculture. In 2008 the salmon aquaculture industry in Australia is worth $260M (AUD) per year. Salmon aquaculture began in the Huon Estuary in the 1980's and production has since increased significantly. The Huon Estuary is nitrogen (N) limited and salmon farming is a significant input of N to this ecosystem. Both industry and government regulators are alert to the potential for eutrophication and increased harmful algal blooms if the assimilative capacity for N of the estuary is exceeded. As part of a larger project on the ecology of the Huon Estuary, this PhD research has two main objectives; firstly to determine whether phytoplankton in the Huon Estuary are using nitrogen that had, primarily, an oceanic source (e.g. nitrate) or was more locally supplied or regenerated (e.g. ammonium and urea) and secondly to examine the physiology of G. catenatum a toxic dinoflagellate that dominates the summer and autumn Huon Estuary phytoplankton biomass in many years. Uptake rates of NO\(_3\)\(^-\), NH\(_4\)\(^+\) and urea were measured on four occasions (28-29 May 2003, 23-24 Sept 2003, 18-19 Nov 2003, and 24-25 Feb 2004) in the Huon Estuary using a \(^{15}\)N tracer technique. Uptake rates were measured at Garden Island and Hideaway Bay in the lower estuary and at 5 and 20 m during the day and also at 5 and 20 m during the night. The mean uptake rates (mean across time of year, site, time of day and depth) for NH\(_4\)\(^+\) (0.13μg N μg chl α h\(^{-1}\) and urea (0.09 μg N μg chl α h\(^{-1}\)1) were 4.5 and 3.2 times higher than the uptake of NO\(_3\)\(^-\) ( 0.3 μtg N μg ch la h\(^{-1}\)). Overall NH\(_4\)\(^+\), NO\(_3\)\(^-\) and urea were responsible for 52, 37.5 and 10.5% of N uptake respectively. Gymnodinium catenatum is a toxic dinoflagellate that blooms periodically in the Huon Estuary and in years that it blooms it dominates the phytoplankton biomass and has caused closure of shellfish farms in the area. Laboratory experiments on effect of temperature and irradiance on growth rate, effect of different nitrogen species on growth rate and preferential uptake of different nitrogen species by G. catenatum were undertaken to better understand the physiology of this species and to test a hypothesis that G. catenatum vertically migrates to access NH\(_4\)\(^+\) at depth during summer. The effect of 12 different temperatures ranging from 11.9-25.2 °C and irradiances from 5-283 gmol photons m\(^{-2}\)s\(^{-1}\) on growth and biochemical composition of G. catenatum. The highest predicted growth rates (>0.2 d\(^{-1}\)) occurred during summer and autumn as might be expected based on observations of bloom dynamics of this species in the Huon Estuary which supports both a summer and autumn bloom in many years. G. catenatum was able to grow using NO\(_3\)\(^-\), NH\(_4\)\(^+\) or urea as its sole nitrogen source. There was no significant difference in growth on any of these nitrogen sources. Preferential uptake of NH\(_4\)\(^+\), NO\(_3\)\(^-\) or urea was examined by growing G. catenatum on a mixture of NO\(_3\)\(^-\), NH\(_4\)\(^+\) and urea. The results clearly showed that NH\(_4\)\(^+\) was taken up first, followed by NO\(_3\)\(^-\) and finally urea. Maximum mean uptake rates were 170, 98 and 30 pg cell\(^{-1}\) hour\(^{-1}\) respectively for NH\(_4\)\(^+\), NO\(_3\)\(^-\) and urea. In addition to the laboratory experiments the nitrogen uptake characteristics of a bloom of G. catenatum was examined at Pelican Island, Southport (30-31/03/2004) nearby the Huon Estuary. Mean urea uptake was greatest (0.045 μgN μg chl α h\(^{-1}\)) followed by NH\(_4\)\(^+\) (0.029 μgN μg chl α h\(^{-1}\)) and the lowest uptake was of NO\(_3\)\(^-\) (0.025 μgN μg chl α h\(^{-1}\)). For G. catenatum growing in the Huon Estuary it seems increasingly apparent that it functions as a nitrogen scavenger. When N concentrations are exhausted, it is able to migrate through the water column seeking whatever form of nitrogen is available.

by Li Yung Yung.<br>Thesis (M.Phil.)--Chinese University of Hong Kong, 2000.<br>Includes bibliographical references (leaves 105-121).<br>Abstracts in English and Chinese.<br>ACKNOWLEDGEMENTS --- p.i<br>ABSTRACT (ENGLISH VERSION) --- p.ii<br>ABSTRACT (CHINESE VERSION) --- p.iv<br>TABLE OF CONTENTS --- p.v<br>LIST OF TABLES --- p.x<br>LIST OF FIGURES --- p.xi<br>LIST OF ABBREVIATIONS --- p.xiii<br>Chapter CHAPTER ONE --- INTRODUCTION<br>Chapter 1.1 --- Seaweed --- p.1<br>Chapter 1.1.1 --- General Introduction --- p.1<br>Chapter 1.1.2 --- Classification and Use of Seaweed --- p.1<br>Chapter 1.1.3 --- Phycocolloids --- p.2<br>Chapter 1.1.4 --- Hong Kong Seaweed --- p.3<br>Chapter 1.1.4.1 --- Sargassum Species --- p.3<br>Chapter 1.1.4.2 --- Padina Species --- p.5<br>Chapter 1.2 --- Source and Production of Alginate --- p.8<br>Chapter 1.2.1 --- Function of Alginate in Seaweed --- p.8<br>Chapter 1.2.2 --- Chemical Structure of Alginate --- p.8<br>Chapter 1.2.3 --- Alginate Production --- p.9<br>Chapter 1.2.4 --- Isolation of Alginate --- p.13<br>Chapter 1.2.5 --- Commercial Methods --- p.13<br>Chapter 1.3 --- Application of Alginate --- p.14<br>Chapter 1.3.1 --- Industrial Application --- p.14<br>Chapter 1.3.2 --- Pharmaceutical Application --- p.16<br>Chapter 1.3.3 --- Food Application --- p.17<br>Chapter 1.3.3.1 --- Uses of Alginate in Food --- p.17<br>Chapter 1.3.3.2 --- Safety --- p.19<br>Chapter 1.4 --- Structure and Function Relationship of Alginate --- p.19<br>Chapter 1.4.1 --- Physico-Chemical Properties --- p.21<br>Chapter 1.4.1.1 --- M/G ratio --- p.21<br>Chapter 1.4.1.2 --- Solution Properties --- p.21<br>Chapter 1.4.1.3 --- Viscosity --- p.23<br>Chapter 1.4.1.4 --- Molecular Weight --- p.27<br>Chapter 1.4.2 --- Functional Properties --- p.27<br>Chapter 1.4.2.1 --- Emulsion --- p.27<br>Chapter 1.4.2.2 --- Gel Properties --- p.27<br>Chapter 1.4.2.3 --- Mechanism of Gelation --- p.29<br>Chapter 1.4.2.4 --- Gel Strength and Syneresis --- p.30<br>Chapter 1.5 --- Physiological Effects --- p.32<br>Chapter 1.5.1 --- Dietary Fibre --- p.32<br>Chapter 1.5.2 --- Minerals --- p.32<br>Chapter 1.6 --- Significance of the Present Study --- p.33<br>Chapter CHAPTER TWO --- MATERIALS AND METHODS<br>Chapter 2.1 --- Seaweed Collection --- p.36<br>Chapter 2.2 --- Sample Preparation --- p.36<br>Chapter 2.3 --- Alginate Extraction --- p.38<br>Chapter 2.3.1 --- Method A --- p.38<br>Chapter 2.3.2 --- Method B --- p.38<br>Chapter 2.3.3 --- Commercial Alginate --- p.39<br>Chapter 2.4 --- Chemical Composition of Alginate --- p.41<br>Chapter 2.4.1 --- Alginate Content --- p.41<br>Chapter 2.4.2 --- Moisture Content --- p.41<br>Chapter 2.4.3 --- Crude Protein Content --- p.41<br>Chapter 2.4.4 --- Ash Content --- p.42<br>Chapter 2.4.5 --- Monosaccharide Composition --- p.42<br>Chapter 2.4.5.1 --- Acid Deploymerisation --- p.42<br>Chapter 2.4.5.2 --- Neutral and Amino Sugar Derivatization --- p.42<br>Chapter 2.4.5.3 --- Determination of Neutral Sugars by Gas Chromatography --- p.43<br>Chapter 2.4.5.4 --- Uronic Acid Content --- p.44<br>Chapter 2.4.6 --- Uronic Acid Block Composition --- p.44<br>Chapter 2.4.6.1 --- "MG, MM and GG Block Determination" --- p.44<br>Chapter 2.4.6.2 --- M/G Ratio Determination --- p.45<br>Chapter 2.4.6.3 --- Phenol-Sulfuric Acid Method --- p.45<br>Chapter 2.5 --- Physico-Chemical Properties of Alginate --- p.46<br>Chapter 2.5.1 --- Viscosity --- p.46<br>Chapter 2.5.1.1 --- Ostwald Viscometer --- p.46<br>Chapter 2.5.1.2 --- Brookfield Viscometer --- p.47<br>Chapter 2.5.2 --- Molecular Weight --- p.47<br>Chapter 2.5.2.1 --- From Intrinsic Viscosity --- p.47<br>Chapter 2.5.2.2 --- Gel Permeation Chromatography-Laser Light Scattering (GPC-LLS) --- p.48<br>Chapter 2.6 --- Functional Properties of Alginate --- p.49<br>Chapter 2.6.1 --- Emulsifying Activity (EA) and Emulsion Stability (ES) --- p.49<br>Chapter 2.6.2 --- Gel Formation --- p.49<br>Chapter 2.6.3 --- Gel Strength and Syneresis --- p.50<br>Chapter 2.6.4 --- Application in Food ´ؤ Fruit Jelly --- p.52<br>Chapter 2.7 --- Data Analysis --- p.53<br>Chapter CHAPTER THREE --- RESULTS AND DISCUSSION<br>Chapter 3.1 --- Proximate Composition of Selected Seaweed --- p.54<br>Chapter 3.1.1 --- Moisture Content --- p.54<br>Chapter 3.1.2 --- Ash Content --- p.56<br>Chapter 3.1.3 --- Crude Protein Content --- p.57<br>Chapter 3.1.4 --- Carbohydrate Content --- p.58<br>Chapter 3.2 --- Chemical Composition of Alginate Extracted from Two Different Methods --- p.58<br>Chapter 3.2.1 --- Percentage Yield --- p.59<br>Chapter 3.2.2 --- Alginate Content --- p.61<br>Chapter 3.2.3 --- Moisture Content --- p.62<br>Chapter 3.2.4 --- Ash Content --- p.62<br>Chapter 3.2.5 --- Residual Protein Content --- p.63<br>Chapter 3.2.6 --- Monosaccharide Composition of Alginate --- p.63<br>Chapter 3.2.7 --- M/G Ratio --- p.66<br>Chapter 3.2.8 --- Summary --- p.69<br>Chapter 3.3 --- Comparative Studies of Physico-Chemical Composition of Alginate from Sargassum and Padina Species --- p.71<br>Chapter 3.3.1 --- Block Composition and M/G Ratio --- p.71<br>Chapter 3.3.2 --- Viscosity --- p.75<br>Chapter 3.3.2.1 --- Intrinsic Viscosity ´ؤ Capillary Viscometer --- p.75<br>Chapter 3.3.2.2 --- Solution Viscosity - Brookfield Viscometer --- p.79<br>Chapter 3.3.2.2.1 --- Effect of Temperature --- p.79<br>Chapter 3.3.2.2.2 --- Effect of Concentration --- p.81<br>Chapter 3.3.2.2.3 --- Shear Thinning and Time Independent Effect --- p.82<br>Chapter 3.3.3 --- Molecular Weight --- p.88<br>Chapter 3.3.3.1 --- From Intrinsic Viscosity --- p.88<br>Chapter 3.3.3.2 --- Gel Permeation Chromatograph-Laser Light Scattering (GPC-LLS) --- p.90<br>Chapter 3.4 --- Comparative Studies of the Functional Properties of Extracted Alginate with Commercial Alginate --- p.93<br>Chapter 3.4.1 --- Emulsifying Activity (EA) and Emulsifying Stability (ES) --- p.93<br>Chapter 3.4.2 --- Gelling Properties --- p.95<br>Chapter 3.4.2.1 --- Effect of Calcium Concentrations --- p.95<br>Chapter 3.4.2.2 --- Gel Strength and Syneresis --- p.97<br>Chapter 3.4.3 --- Application in Food --- p.99<br>Chapter CHAPTER FOUR --- CONCLUSIONS --- p.103<br>REFERENCES --- p.105<br>RELATED PUBLICATION --- p.120

Includes bibliographical references (leaves 235-258).<br>xxi, 274 leaves, [8] leaves of plates : ill. (chiefly col.), map 30 cm.<br>Estimates the primary production and investigates the photosynthetic performance of temperate turfs at West Island, off the coast of South Australia. These communities play a fundamental role in reef ecology, being the main source of food for grazers, both fishes and invertebrates. Turfs also have an important function in benthic algal community dynamics, being the first colonizers on disturbed and bare substratum.<br>Thesis (Ph.D.)--University of Adelaide, Dept. of Environmental Biology, 2002

This study investigated the effects of nitrogen and phosphorus on the growth and ecophysiology of a number of dominant species of macroalgae at a site in Port Phillip Bay (PPB), a large shallow water marine embayment located on the central southern coast of Victoria, Australia. This thesis investigated the physiological processes (i.e. photosynthesis, growth, nutrient uptake) of three species of macroalgae, Hincksia sordida (Harvey) Clayton (Phaeophyta), Polysiphonia decipiens Montague (Rhodophyta) and Ulva sp. (Chlorophyta) in response to a range of environmental regimes.