published_or_final_version / abstract / Biological Sciences / Doctoral / Doctor of Philosophy

A busca por segurança energética tem feito com que a maioria dos países empenhe-se na busca por fontes alternativas de energia, procurando mitigar problemas econômicos, sociais e ambientais. Espera-se que a biomassa, disponível de forma geograficamente dispersa, venha a tornar-se um dos principais recursos renováveis na produção de alimentos, materiais, produtos químicos, combustíveis e energia. Nesse cenário, o desenvolvimento de biorrefinarias representa a chave para uma produção integrada, combinando rotas de conversões químicas, bioquímicas e termoquímicas, no processamento da biomassa, visando à utilização otimizada dos recursos disponíveis. As biorrefinarias com estruturas descentralizadas são uma alternativa à centralização de produções em grandes plantas industriais e à monocultura, pois utilizam biomassas disponíveis regionalmente, integram sistemas de produção, potencializando os recursos locais, reduzem custos com logística e impactos ambientais, além de melhorarem a distribuição da renda. O presente trabalho apresenta uma revisão da disponibilidade de biomassa no Brasil e no mundo, com especial interesse no aproveitamento de microalgas e resíduos orgânicos, dos principais produtos de interesse e conceitos de biorrefinarias existentes. O objetivo é discutir qual é o conceito que melhor se adapta às necessidades do cenário brasileiro, bem como propor um modelo com estrutura descentralizada em duas ou três etapas, visando à otimização de um processamento sustentável de biomassa para obtenção de vários produtos comerciáveis e energia, além de delinear diretrizes para investimentos na área. É apresentada uma metodologia para a tomada de decisões na concepção e análise de viabilidade do projeto conceitual de uma biorrefinaria, considerando-se também as restrições de ordem ecológica, econômica e tecnológica. Como estudo de caso, é proposta uma biorrefinaria a partir de microalgas. A escolha por microalgas como matéria-prima é baseada nas vantagens que sua utilização apresenta frente à de outras biomassas, dentre elas cita-se a capacidade de produção rápida e durante todo o ano, a captura do CO2 necessário ao seu crescimento, a necessidade de menos água do que plantas terrestres, são cultiváveis em água salobra e terras não aráveis, apresentam elevado teor de óleo, seus nutrientes podem ser obtidos a partir de águas residuais, sua composição bioquímica pode ser modulada por diferentes condições de crescimento e são capazes de produção fotobiológica de bio-hidrogênio. Das alternativas de rotas possíveis para seu processamento, são apresentadas duas que apontam como sendo as mais promissoras: o uso da microalga como substrato de algum outro microorganismo, visando à obtenção de compostos com maior valor agregado, tais como biopolímeros, e o uso do processo de pirólise rápida para obtenção de bio-óleo, que deve ser processado posteriormente, visando à especificação em biocombustível. As vantagens da otimização do cultivo, colheita, rotas viáveis de processamento e a análise do potencial econômico desse modelo, indicam uma excelente oportunidade para obtenção de um espectro de produtos de alto valor agregado e energia e um grande potencial de aplicação. / The search for energy security has been doing with that most of the countries strive to seek alternative sources of energy that allows mitigating economic, social and environmental problems. It is expected that biomass, available on a geographically dispersed way, will become one of the major renewable resources for food, materials, chemicals, fuels and energy production. Additionally, the development of biorefineries represents the key to an integrated production, combining chemical, biochemical and thermochemical conversion routes for biomass processing, aiming the optimized use of available resources. Biorefineries with decentralized structure are an alternative to the centralization of production in large industrial plants and to the monoculture because it uses biomass regionally available, integrates production systems potentiating local resources, reduces logistics costs and environmental impacts, as well as improves income distribution. This work presents a review of: i) biomass availability in Brazil and in the world, focus on the use of microalgae and organic wastes; ii) the main products of interest; and; iii) concepts of existing biorefineries. The objective is to discuss which one is the concept that better fits the needs of the Brazilian scenario and propose a new model with decentralized structure in two or three stages, seeking to an optimized and sustainable biomass processing to obtain various marketable products and energy, and delineate guidelines for investment in the area. It is presented a methodology for making the conceptions decisions and feasibility analysis of the conceptual design of a generic refinery, considering also the ecological, economic and technological constraints. As a case study, it is proposed a biorefinery from microalgae. The choice of microalgae as a raw material is based on the advantages that their use offers over the others biomasses. Among the advantages it can be cited the ability for rapid production and during all the year, the capture of CO2 for its growth, the need for less water than land plants, they can be cultivated in brackish water and non-arable land, have a high oil content, nutrients can be obtained from waste water, their biochemical composition can be modulated by different growth conditions and are capable of producing photobiology bio-hydrogen. Of the possible alternative routes for microalgae processing, are presented two as the most promising: the use of microalgae as a substrate for some other microorganism in order to obtain compounds with high added value, biopolymers as an example, and use the process of fast pyrolysis for obtaining bio-oil, which will be processed later in order to specify biofuels. The advantages of the cultivation optimization, harvesting, processing viable routes and analyzing the economic potential of this model indicate an excellent opportunity to obtain a spectrum of value-added products and energy and its great potential for application.

Memoria para optar al Título Profesional de Ingeniero Forestal / Didymosphenia geminata (Didymo) es una microalga bentónica declarada plaga en el año 2013 por la Subsecretaria de Pesca. Esta tiene una amplia distribución en Chile, encontrándose desde la Región del Biobío hacia el sur del país. Las floraciones de esta especie, ocupan una alta proporción de la superficie de los ríos, alcanzando incluso un 100% de cobertura. Debido a los altos costos ambientales y económicos que provoca la presencia de la especie, es necesario determinar su potencial de expansión en el país y, de esta manera poder enfocar los esfuerzos para su control en las zonas más críticas. Es así como el objetivo de esta memoria fue estudiar la distribución actual y potencial de la microalga D. geminata en Chile. De este modo, mediante el uso de distintas variables bioclimáticas, ambientales y geográficas fue posible determinar la distribución potencial de la especie, encontrando que ésta tiene alta probabilidad de expandir su presencia por varias regiones del país. Este procedimiento se llevó a cabo con el algoritmo MAXENT, el cual fue escogido gracias a las buenas predicciones que ha presentado para esta especie con respecto a otros métodos. Por otro lado, se determinó que las variables que más incidencia tienen en la presencia de la especie son la variabilidad estacional, flujo acumulado y la precipitación del periodo más seco del año. / Didymosphenia geminata (Didymo) is a benthic microalgae, declared as a plague in 2013 by the Subsecretaria de Pesca. This species has a wide distribution in Chile, from Biobío Region towards the south of the country. The bloom of this species, occupy a high proportion of the rivers surface, reaching even a 100% of coverage. Due to the high environmental and economic costs caused by the presence of the species, it is necessary to determine its potential expansion in the country and, in this way, it being able to focus all efforts for controlling it in the most critical areas. Thus, the objective of this report was studying the current and potential distribution of D. geminata microalgae in Chile. Therefore, through the use of various bioclimatic, environmental and geographic variables it was possible to define the potential distribution of the species, concluding that it has a high probability of expanding its presence in several regions of the country. This procedure was carried out using the MAXENT algorithm, which was chosen due to the good predictions presented for this species with respect to others methods. On the other hand, it was determined that the variables which are most incidental in the presence of the species are seasonal variability, and the flow and precipitation accumulation in the driest quarter of the year. / Diciembre 2018

Graduation date: 1999

Graduation date: 1995

Microalgae are an important potential feedstock for biodiesel production. Understanding the physiology of lipid biosynthesis in microalgae is pivotal to microalgal aquaculture management. A freshwater green microalga strain, UTEX 2219-4, was isolated from UTEX 2219 which was reported containing two strains. Its ITS sequences are closely related to those in the family of Scenedesmaceae in the GenBank. Nitrogen starvation, salt stress and osmotic stress greatly enhanced lipid biosynthesis in this strain, while combination of nitrogen deficiency and osmotic stress had the most dramatic effect. Chloroplast was condensed and photosynthesis efficiency declined about 50% after 3 days of nitrogen starvation. Chlorophyll degradation followed the same trend but was more severe than the reduction of photosynthesis efficiency. Oil body formation was not observed in the cells kept in the dark under nitrogen starvation, suggesting photosynthesis rather than autophagy is the major player in oil body formation. Under non-saturation levels of light intensities coupled with nitrogen starvation, the oil body formation under 150 £gmol/m2s light intensity was more efficient than that under 75 £gmol/m2s. DCMU blocked photosynthesis as well as oil body formation, supporting that the energy for oil body formation was mostly from photosynthesis rather than autophagy during nitrogen starvation.

A new species of reddish-orange pigment-producing microalga was isolated from a shallow pond in tropical Taiwan. Morphological and molecular evidence including meridional ribs on the cell wall, pigment production, and 18S rDNA sequence analysis suggest that this microalga is a species in the genus Coelastrella. Salt stress accelerated biosynthesis of the reddish-orange pigments, and large quantity of oil accumulated as the cells stressed under nutrient deficiency. This microalga could sustain 45 ¢XC for more than 8 hours indicated by the stability of its chlorophylls, which is a necessary trait for large scale outdoor cultivation using photobioreactors in tropical areas. The reddish-orange pigments could be separated into many fractions by HPLC, and signals from carotenoids were detected in a few fractions using NMR, suggesting these pigments may function as antioxidants among other roles.

Benthic microalgae (BMA) living within the surface sediment of salt marshes are highly productive organisms that provide a significant proportion of organic carbon inputs into estuarine systems. BMA secrete extracellular carbohydrates in the form of low molecular weight carbohydrates and extracellular polymeric substances (EPS) as they migrate within the sediment. EPS plays an important role in the structure and function of BMA biofilms in shallow-water systems as EPS affects habitat structure, stabilizes the sediment, reduces sediment erosion, and is a carbon source for organisms. This study looked at the effect of nutrients and carbohydrate additions on BMA biomass, bacterial biomass, carbohydrate production, and glycosidase activity in the surface 5 mm of intertidal sediment in a subtropical salt marsh (Galveston Bay, Texas). Nitrogen and phosphorus were added to cores collected from the salt marsh and incubated in the lab over four days. Very little change was seen in the biomass of the benthic microalgae or in the different carbohydrate fractions with the added nutrients. The mean chlorophyll a concentration was 13 +/- 5 ug g-1 sediment, the mean saline extractable carbohydrate concentration was 237 +/- 113 ug g-1 sediment, and the mean EPS concentration was 48 +/- 25 ug g-1 sediment. The chlorophyll a and saline extractable carbohydrate concentrations initially decreased over the first 24 hours, but then increased over the rest of the experiment, indicating a possible species compositional shift in the BMA. With no major response with nutrient additions, it is likely that a different environmental factor is limiting for the growth of the benthic microalgae, and therefore the production of sEPS, in this salt marsh. A series of experiments was conducted in situ by adding glucose, alginic acid, and phosphorus to sediment within experimental plots. Samples were taken periodically over three to seven days to determine the biomass of the microbial community, enzyme activities and kinetics, and changes in the concentrations of several sediment carbohydrate pools. u-glucosidase activities (15 +/- 3 nmol g-1 h-1) were significantly higher than u-xylosidase (6 +/- 2 nmol g-1 h-1) and u-galactosidase (8 +/- 2 nmol g-1 h-1) activities within the sediment, and there was no suppression of u-glucosidase activity measured with the glucose addition. These data represent the first measurement of u- xylosidase and u-galactosidase activity in intertidal sediment dominated by BMA. Although preliminary experiments suggested a possible phosphorus limitation within the sediment, there was little change in the bacteria abundance or the benthic microalgae biomass when phosphorus was added in situ. This study begins to illustrate the dynamics of carbohydrate production and loss in this salt marsh, and the ability for the microbial community in the salt marshes of Galveston Bay to adjust to the nutrient and carbohydrate treatments.

Plankton communities are important members of the food web in coastal systems and are regulated by top-down and bottom-up controls. This study examined the influence of bottom-up controls, such as physicochemical factors, and top-down controls, such as predation, on the plankton communities in South Bay, Texas. Microalgal photopigments were ascertained by high-performance liquid chromatography (HPLC) to determine the relative abundances of major algal classes. Zooplankters were identified to the lowest possible taxon and enumerated. No spatial trends were observed for the physicochemical factors. The northern bay sections exhibited significantly higher phytoplankton and microphytobenthic diatom biomass, probably due to their proximity to the bay inlet. Copepod, gastropod veliger and brachyuran zoea abundances were also higher in this area, albeit insignificantly. The southern bay sections experienced significantly higher cyanobacterial, euglenophyte and chlorophyte biomass, and polychaete larval abundances. Total zooplankton and nauplii abundances were also higher in the southern areas, albeit insignificantly. Sampling the inaccessible areas of the bay in the future may reveal spatial variability among the physicochemical factors which could be influencing the distribution of plankton. Temporal variation for the physicochemical factors followed a typical trend for subtropical climates and influenced the seasonality of the plankton communities. Phytoplankton biomass peaked in February, August and October but these maximums were not significantly different from the other months sampled. Microphytobenthic biomass peaked during the summer months, while diatom biomass also peaked in February. Zooplankton abundances peaked in October, while nauplii and polychaete larvae also peaked in February. Relationships between wind speed, turbidity and the microalgal pigments were assessed to determine if wind-induced resuspension influenced the location of the major algal classes within the water column compared to the sediments. Wind speed and turbidity were directly related to each other, albeit insignificantly. Some phytoplankton and microphytobenthos were considered tychopelagic because wind-induced resuspension increased their biomass in the water column compared to the sediments. The physicochemical factors exerted bottom-up control of plankton community dynamics in this study, while top-down controls, such as predation, require further investigation. Future studies should focus on which of these controls have more influence on plankton community dynamics in South Bay.

Microalgae have been proposed as a potential feedstock for biofuel production, and as a result, interest in the biology of these organisms has intensified. These organisms also synthesize lipids that are vital to human health and nutrition. Stress has been shown to have an effect on lipid composition and gene expression in microalgae, but many studies have focused on the effects of abiotic stressors. The purpose of this study was to investigate the effect of biotic stress on lipid-related gene expression in Phaeodactylum tricornutum, a model species of microalgae. The source of biotic stress used in this study was 2E,4E/Z-decadienal, a diatom-derived oxylipin that has been shown to function as a stress signal among diatoms. Real-time RT-qPCR analysis revealed that expression of a patatin-like phospholipase was significantly decreased in decadienal-treated cultures as compared to a solvent control. The expression of a delta-9 desaturase gene believed to be responsible for production of 16:1 fatty acids was increased by a factor of 12. FabI, a gene involved in fatty acid biosynthesis, and PtD5a, which codes for an ER-localized desaturase, were both down-regulated in cells exposed to decadienal. However, changes in expression were only shown to be significant for the patatin-like phospholipase gene. Increased expression of the delta-9 desaturase gene may be a protective mechanism against infection from pathogens, since 16:1 fatty acids have been shown to have antibacterial properties. Regulation of membrane desaturation may also serve to stabilize photosynthetic membranes during times of stress. The down-regulation of the phospholipase gene was surprising, since the release of fatty acids from membrane lipids for oxylipin production is a common response to stress. It is recommended that this experiment be improved upon and expanded in order to determine whether the results obtained are reproducible and how these changes in gene expression correlate with physiological effects. / text