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No. of bitstreams: 1 DissECC.pdf: 1201673 bytes, checksum: c96ddf2834cfea8ddcc495673a274880 (MD5) Previous issue date: 2016-03-14 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / The cement industry, closely associated with the global warming question, accounts for significant emissions of CO2 and other air pollutants, such as SO2 and NO2 in the atmosphere. In search for ways to mitigate the atmospheric CO2, we performed semicontinuous cultures of Chlorella sorokiniana under phototrophic conditions to test the effect of a flue gas simulation (18% CO2, 9% O2, 300 ppm NO2 and 140 ppm SO2). This was provided once a day in six serial experiments, in which the exposure to the gas was increased through the increase of bubbling time. A constant flow rate allowed us to calculate the total volume of gas introduced into the system each day (0.1, 0.3, 0.8, 1.5, 6 and 48 L). Air-CO2 (18%) was used as control and its bubbling time was pHregulated. Culture medium acidification led to suboptimal growth conditions that affected cell density, photosynthetic activity, cell viability and the biochemical composition of C. sorokiniana. Compared to control, the specific growth rate decreased by 17 and 3,9% in cultures that received 6 and 48 L gas d-1, respectively. The pulseamplitude modulated (PAM) fluorometry was used for culture evaluation. It revealed low maximum quantum yield (ΦM 0.40) and operational quantum yield (Φ'M 0.47) values one day after 48 L gas bubbling. Light saturation curves confirmed the negative effects of long-time gas simulation stress. On the other hand, quenching analysis indicated an increase in photochemical light use and low values of non hotochemical quanching (qN and NPQ). Exposure of the cells to the flue gas simulation resulted in lower cell viability compared to control. Biochemical analysis showed that 6 and 48 L gas d-1 significantly increased protein content by 75% and 154%, respectively; total carbohydrates also increased in the presence of the gas, 148% and 195%, respectively. Despite the physiological changes, C. sorokiniana resisted suboptimal growth conditions imposed by the gas, supporting its vigorous nature and relevance in biotechnological aplications with flue gases. / Inserida na problemática do aquecimento global, a indústria de cimento é uma das que mais contribui para emissão de CO2 e de poluentes como SO2 e NO2 na atmosfera. Visando alternativas para mitigação desses gases, que são oriundos principalmente de processos de combustão, foram testados seis cultivos fototróficos semicontínuos de Chlorella sorokiniana, para avaliar o efeito de diferentes volumes (0,1; 0,3; 0,8; 1,5; 6 e 48 L d-1) de uma simulação gasosa composta por CO2 (18%), O2 (9%), NO2 (300 ppm) e SO2 (140 ppm). Os volumes variaram conforme o aumento do tempo de borbulhamento diário dos gases nos cultivos. O tratamento controle, composto por ar sintético e CO2 (18%), teve seu tempo de borbulhamento definido pela variacão de pH do meio. O fornecimento da simulação gasosa resultou na acidificação do meio de cultura e afetou a densidade celular, a atividade fotossintética, a viabilidade celular e a composição bioquímica de C. sorokiniana. Comparadas ao controle, as exposições diárias de 6 e 48 L gás d-1 reduziram a taxa específica de crescimento em 17 e 39%, respectivamente. Por meio da fluorescência de amplitude modulada (PAM), verificamos baixos valores de rendimento quântico máximo (ΦM 0,40) e operacional (Φ’M 0,47) um dia após o primeiro borbulhamento de 48 L gás. Curvas de saturação de luz confirmaram os efeitos negativos do estresse prolongado à mistura gasosa. A análise de decaimento da fluorescência da clorofila, por sua vez, indicou um aumento da energia luminosa direcionada à fotoquímica da fotossíntese (qP) e baixos valores de dissipação não fotoquímica da energia luminosa (qN e NPQ). A simulação gasosa resultou, ainda, em menor viabilidade celular, se comparada ao controle. Pelas análises bioquímicas, constatamos que 6 e 48 L gás d-1 levaram a um aumento significativo do conteúdo proteico de 75% e 154%, respectivamente; os mesmos tratamentos também aumentaram a quantidade de carboidratos totais em 148% e 195%. Apesar das alterações fisiológicas, C. sorokiniana resistiu às condições subótimas de crescimento, o que comprova sua robustez e relevância em aplicações biotecnológicas envolvendo gases de combustão.

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<p>Owing to the rise in fossil fuel prices, overall energy security concerns, and the current push towards green engineering; renewable and green fuels have seen an increase in interest in recent years. Two notable technologies in this green movement are the production of biodiesel from microalgae and the production of biogas from anaerobic digestion of waste biomass. Production of biodiesel from microalgae was studied extensively in the 80s through the early 90s and found to be economically infeasible given the technology of the time. However, recent literature has suggested that one possible method to improve the feasability of the process would be to combine it with an anaerobic digestor to provide nutrient and biomass recycling. For such a system, having accurate models of each process would be highly advantageous for optimal design and control. To this end this thesis moves towards this overall goal by examining and modelling the anaerobic digestion of the microalgae <em>Chlorella vulgaris</em>.</p> <p>Starting with a set of experimental data (anaerobic digestion of <em>Chlorella vulgaris</em>) provided by LBE-INRA, the minimum number of kinetic equations needed to predict the data are found using principal component style analysis. This number is found to be two to three reactions. Using this as a basis for model development, a mass balance model is developed around both two and three reaction cases. To date there is very little literature on the modelling of anaerobic digestion of microalgae and so kinetic laws are selected from the general anaerobic digestion models ``Anaerobic Digestor Model 1'' (ADM1) and ``Acidogenesis/Methanogenisis Model'' (AM2). Given that the kinetic laws were derived from general literature, model fitting is a must. To faciliate this process a novel systematic parameter identification procedure to locate identifiable parameter subsets within each model is presented. Applying this novel procedure to the provided data is seen to lead to promising identification results. Through these identification trials it is shown that the three reaction model best captures the dynamics of the system. This three reaction model serves as the basis for subsequent steady state optimality and sensitivity analysis. From these efforts it is shown that the predicted optimal curves match literature data very well but uncertainty in certain key parameter estimates lead to highly sensitive model predictions (and therefore low confidence). This leads to the conclusion that the developed model is capable of predicting the kinetics of <em>Chlorella</em> digestion but additional trials are needed to further refine the model fitting results.</p> <p>Coupling an anaerobic digester to a microalgal culture is currently considered one of the most promising avenues towards the production of renewable bioenergy, either in the form of biodiesel or biogas. Accurate mathematical models are crucial tools to assess the potential of such coupled biotechnological processes and help optimize their design, operation and control. This paper focuses on the compartment of anaerobic digestion of microalgae. Using experimental data for the anaerobic digestion of <em>Chlorella vulgaris</em>, a grey-box model is developed that allows good prediction capabilities and retains low complexity. The proposed methodology proceeds in two steps, namely a structural and a parametric identification steps. The fitted model is then used to conduct preliminary optimization for the production of biogas from <em>Chlorella vulgaris</em>. The results provide some insight into the potential for bioenergy production from the digestion of microalgae and, more generally, the coupled process.</p> / Master of Applied Science (MASc)

Dans le cadre de la protection de l'environnement, l'eau et l'atmosphère sont deux milieux particulièrement surveillés. La réglementation impose la détection et le dosage d'une liste de produits mais les besoins analytiques sont considérables. Les méthodes biotechnologiques présentent l'avantage d'indiquer l'impact du produit sur la matière vivante. Ces méthodes sont mises en œuvre au laboratoire ou sur site. La détection d'une toxicité globale ou ciblée, <i>in situ</i> et en temps réel, est privilégiée. Les biocapteurs sont des outils répondant à cette attente. Au cours de l'étude, deux biocapteurs, l'un ampérométrique, l'autre fluorimétrique sont conçus. Tous deux intègrent le même biorécepteur, <i>Chlorella vulgaris</i>. Une nouvelle méthode d'immobilisation des micro-algues est mise au point. Elle permet d'obtenir des lots de membranes reproductibles, fonctionnelles 7 jours, donnant une réponse <i>in vivo</i> à valeur statistique, réutilisables et conservables. Des essais conduits sur des thylakoïdes extraits des cellules végétales ne donnent pas d'aussi bons résultats. L'association des membranes algales avec les deux types de transducteur montre que <i>Chlorella vulgaris</i> est sensible à des produits divers : herbicides, métaux, solvants. Les limites de détection pour des herbicides sont particulièrement basses, inférieures aux normes. Pour plusieurs produits testés, des courbes d'étalonnage sont données. Les deux types de biocapteur fonctionnent en milieu aqueux, en mode batch et en mode flux. Le biocapteur de fluorescence algale à fibres optiques possède des qualités de reproductibilité et de maniabilité plus intéressantes que le biocapteur ampérométrique. Le biocapteur de fluorescence est testé sur des lixiviats de bois, solutions naturelles complexes ; le biocapteur ampérométrique est adapté à l'utilisation en phase gazeuse, il détecte alors le méthanol vapeur et le perchloroéthylène en aérosol. Une autre partie de l'étude consiste à caractériser l'impact des toxiques sur <i>Chlorella vulgaris</i>. L'analyse est conduite en microscopie électronique à balayage et en microscopie optique couplée à l'analyse d'images. Ces deux méthodes originales restent à perfectionner. Les résultats de nos essais préliminaires semblent encourageants pour la détection du produit toxique et l'accès à son mécanisme d'action.

Tesis por compendio / [ES] Las microalgas son organismos unicelulares fotosintéticos microscópicos que se encuentran en gran variedad de ambientes y son muy eficientes en la transformación de energía solar en biomasa. Los estudios realizados hasta el momento hacen referencia a posibles beneficios de la incorporación de microalgas en la dieta, por la mejora del sistema cardiovascular, las propiedades adelgazantes y energizantes, capacidad antioxidante, o la reducción del colesterol y los triglicéridos. La forma más habitual de consumir las microalgas es como suplemento dietético en forma de tabletas, cápsulas o polvo. La incorporación de biomasa de microalgas en productos tradicionales se ha enfrentado al reto de la aparición de colores verdes fuertes, así como su consistencia pulverulenta que puede afectar la textura y percepción del producto. Todos estos aspectos constituyen las principales áreas de mejora para conseguir un mayor grado de aceptación de productos con microalgas, y son la base del reto de este proyecto. El objetivo de la presente tesis doctoral fue el desarrollo de nuevos productos alimentarios incorporando las propiedades nutricionales de la biomasa de microalgas, incrementando o mejorando con ello, las propiedades nutricionales del alimento original. Para conseguir este objetivo se evaluaron a nivel fisicoquímico, reológico y textural, la incorporación de diferentes especies de microalgas (Arthrospira platensis (Spirulina), Chlorella vulgaris, Dunaliella salina y Nannochloropsis gaditana) en distintas matrices alimentarias (productos horneados, emulsiones y extrusionados). Por otra parte, se evaluó y caracterizó la incorporación de las microalgas utilizando diferentes tecnologías como la impresión 3D o la extrusión. Además de cómo afecta la incorporación de las microalgas a los productos obtenidos, se evaluaron los aspectos nutricionales de su incorporación, en cuanto al aporte de minerales y su biodisponibilidad. Las propiedades reológicas de las masas y emulsiones enriquecidas con microalgas (Spirulina, Chlorella y Dunaliella) indicaron que su comportamiento viscoelástico fue modificado y mejorado, mostrando características aptas para este tipo de productos. La adición de microalgas (Spirulina y Chlorella) a las masas utilizadas para la impresión 3D de galletas y snacks, permitió una mejor extrusión o impresión de éstas, obteniendo muestras impresas en 3D de forma cilíndrica, más precisas en cuanto a sus dimensiones con respecto a la estructura cilíndrica diseñada. Además, las muestras impresas presentaron mayor estabilidad y resistencia, antes y después del proceso de horneado comparadas con la muestra control. Para los productos horneados, tanto para las rosquilletas como los snacks impresos en 3D, la adición de microalgas (Spirulina y Chlorella) permitió mayor estabilidad en términos de textura. Ligeros cambios en los parámetros fisicoquímicos y de expansión se produjeron por la adición de Spirulina y Chlorella en los productos extrusionados. Además, los extrusionados enriquecidos con Nannochloropsis, mostraron parámetros similares a los de la muestra de control. Todos los productos presentaron colores luminosos y apariencias innovadoras y atractivas. En cuanto a los minerales, se observó un aumento de P, K, Ca, Na, Mg, Fe y Se con la adición de Spirulina y Chlorella, junto con el aumento de la concentración adicionada de microalgas. Siguiendo la normativa sobre etiquetado nutricional de los alimentos, el enriquecimiento con microalgas en rosquilletas se puede clasificar como un alimento "rico en hierro (Fe)". De igual forma, las rosquilletas y galletas enriquecidas con microalgas pueden considerarse un alimento "alto en selenio (Se)". Además, la incorporación de Spirulina y Chlorella en las formulaciones de galletas, permitió una mayor bioaccesibilidad del contenido de P, K, Ca, Mg, Fe, Zn y Se para la absorción en el cuerpo comparado con las muestras control. / [CAT] Les microalgues són organismes unicellulars fotosintètics microscòpics. Són molt eficients a l'hora de transformar l'energia solar en biomassa. Els estudis realitzats fins ara fan referència a possibles beneficis de la incorporació de microalgues en la dieta per produir una millora del sistema cardiovascular, per presentar propietats per aprimar i donar energia, per mostrar capacitat antioxidant o per afavorir una reducció del colesterol i els triglicèrids. La forma més habitual de consumir microalgues és com a suplement dietètic en forma de tauleta, càpsula o en pols. La incorporació de biomassa de microalgues en productes tradicionals s'ha afrontat al repte de l'aparició d'un color verd fosc i d'una consistència polsosa que pot afectar a la textura i, per tant, a la percepció del producte. Aquests aspectes constituïxen les principals àrees de millora per aconseguir un major grau d'acceptació de productes amb microalgues i són la base del repte d'aquest projecte. L'objectiu d'aquesta tesi doctoral és el desenvolupament de nous productes alimentaris que incorporen les propietats nutricionals de la biomassa de microalgues, de manera que s'incrementen o es milloren les propietats nutricionals de l'aliment original. Per aconseguir aquest objectiu s'avaluaren a escala fisicoquímica, reològica i de textura la incorporació de diferents espècies de microalgues (Arthrospira platensis (Spirulina), Chlorella vulgaris, Dunaliella salina i Nannochloropsis gaditana) en diferents matrius alimentàries (productes fornejats, emulsions i extrudits). D'altra banda, s'avaluà i caracteritzà la incorporació de les microalgues utilitzant diferents tecnologies com la impressió en 3D o l'extrusió. A banda de valorar com afecta la incorporació de microalgues als productes elaborats, s'avaluaren els aspectes nutricionals, pel que fa a l'aportació i biodisponibilitat de minerals. Les propietats reològiques de les masses i emulsions enriquides amb microalgues (Spirulina, Chlorella i Dunaliella) indicaren que el seu comportament viscoelàstic fou modificat i millorat, de tal manera que mostrà característiques aptes per aquest tipus de productes. L'addició de microalgues (Spirulina i Chlorella) en les masses utilitzades per a la impressió 3D de galetes i snacks permeté una millor impressió, ja que s'obtingueren mostres impreses de forma cilíndrica amb unes dimensions més precises respecte a l'estructura cilíndrica dissenyada. A més, les mostres impreses presentaren una major estabilitat i resistència abans i després del procés de fornejat en comparació amb la mostra control. Respecte als productes fornejats, l'addició de microalgues (Spirulina i Chlorella) a les rosquilletes i els snacks impresos en 3D permeté una major estabilitat en termes de textura. Lleugers canvis als paràmetres fisicoquímics i d'expansió es produïren per l'addició d'Spirulina i Chlorella en els productes extrudits. A més, els extrudits que foren enriquits amb Nannochloropsis mostraren paràmetres similars als de la mostra control. Tots els productes presentaren colors lluminosos i aparences innovadores i atractives. Pel que fa als minerals, s'observà un augment de P, K, Ca, Na, Mg, Fe i Se quan s'afegí Spirulina i Chlorella, directament relacionat amb l'augment de la concentració de microalgues. Seguint la normativa sobre etiquetatge nutricional dels aliments, l'enriquiment amb microalgues en rosquilletes ens permet classificar-les com a aliment "ric en ferro (Fe)". De la mateixa manera, les rosquilletes i galetes enriquides amb microalgues poden considerar-se un aliment "alt en seleni (Se)". A més a més, la incorporació de Spirulina i Chlorella en les formulacions de galetes, permeté una major bioaccessibiltat del contingut de P, K, Ca, Mg, Fe, Zn i Se comparat amb les mostres control. / [EN] Microalgae are microscopic unicellular and photosynthetic organisms that can be found in a wide variety of environments. These microorganisms are very efficient when transforming solar energy into biomass, due to their cellular structure, which is completely submerged in an aqueous medium, forming an adequate surface for the exchange of nutrients and gases. Microalgae compounds are now known to exhibit cardioprotective, immunomodulatory, anti-proliferative, anti-inflammatory, cognitive, neurobehavioral and antimicrobial properties, amongst others. Researchers have shown possible benefits of the incorporation of microalgae in the diet so far. The most common way to consume microalgae is as a dietary supplement in the form of tablets, capsules or powder. The incorporation of microalgae biomass in traditional products has faced the challenge of the appearance of strong green colours, as well as its powdery consistency that can affect the texture and perception of the product. All these aspects constitute the main areas for improvement the development of microalgae-based products, and they are the challenges faced of this project. The main objective of this PhD Thesis was the development of novel food products incorporating the nutritional properties of microalgae biomass, thereby increasing or improving the nutritional properties of the original food matrix. To achieve this goal, the effect of the addition of different species of microalgae (Arthrospira platensis (Spirulina), Chlorella vulgaris, Dunaliella salina and Nannochloropsis gaditana) on the physicochemical, rheological and textural properties of different food matrices (baked products, emulsions and extrudates) was evaluated. Furthermore, the effect of the incorporation of microalgae using different technologies such as 3D printing or extrusion to obtain food products was studied. In addition, how the incorporation of microalgae affects the nutritional aspects of the food products in terms of the contribution of minerals and their bioavailability was also evaluated. The rheological properties of doughs, batters and emulsions enriched with microalgae (Spirulina, Chlorella and Dunaliella) indicated that their viscoelastic behaviour was modified and improved, showing characteristics suitable for this type of products. The addition of microalgae (Spirulina and Chlorella) to the doughs and batters used for the 3D printing of cookies and snacks, allowed a better extrusion or printing behaviour. This allowed obtaining cylindrical 3D printed samples, more precise in terms of their dimensions with respect to the designed cylindrical structure. In addition, the 3D microalgae-printed sample structures presented greater stability and resistance, before and after the baking process compared to the control sample. For baked products, both for breadsticks and 3D printed snacks, the addition of microalgae (Spirulina and Chlorella) allowed greater stability in terms of texture. Slight changes in the physicochemical and expansion parameters were produced by the addition of Spirulina and Chlorella in the extruded products. In addition, the extrudates enriched with Nannochloropsis, showed similar parameters to those of the control sample. Microalgae-enriched obtained products showed bright colours with appealing appearances. Regarding minerals, an increase in P, K, Ca, Na, Mg, Fe and Se was observed with the addition of Spirulina and Chlorella, along the increase of concentration of microalgae addition. Following the regulations on nutrition labelling for food stuffs, breadstick enrichment with microalgae are a food "high in iron (Fe)" In the same way, breadsticks and cookies enriched with microalgae can be considered a "high in selenium (Se)" food. Going a step further, Spirulina and Chlorella vulgaris incorporation in cookie formulations allowed for greater bioaccessibility of P, K, Ca, Mg, Fe, Zn, and Se content for absorption in the body than control cookies. / Uribe Wandurraga, ZN. (2020). Microalgae as novel ingredients for the formulation of food products [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/158743 / Compendio