Realizou-se um estudo do processo descontínuo alimentado de fermentação, buscando encontrar as melhores condições de produção do ácido graxo gama-linolênico. Empregou-se como fonte de nitrogênio no meio de fermentação a uréia, que foi adicionada de forma exponencialmente crescente e como inóculo usou-se uma cepa de Spirulina platensis. Utilizou-se planejamento experimental e aplicou-se a metodologia de superfície de resposta, o qual permitiu a obtenção de equações que relacionaram as variáveis dependentes, como produtividade em células, conversão de nitrogênio em células, concentração máxima atingida de biomassa, porcentagem relativa de produção de ácido graxo gama-linolênico. com as variáveis independentes tempo de alimentação (Ta) e massa total de uréia adicionada (MUT). Os melhores resultados obtidos foram os seguintes: Px=117,26 mg/L.dia; Yx/n= 4,9; Xmax= 1759 mg/L e porcentagem de ácido graxo γ-linolênico na fração lipídica de 23,5 %, com 14 dias (Ta) e 1750 mg (MUT). / A Study on the fed-batch fermentation process was made seeking for the best Gamma-linolenic production condition. Mineral media was employed with urea as nitrogen source that was add with exponentially increasing feeding rates and Spirulina platensis as inoculum. Experimental design and a Response Surface Methodology were applied. It was composed by an experimental design which allowed a reduced number of the experiments and by the analysis of the results using equations which combined the interest variables dependent as: Productivity, best Gamma-linolenic production condition, Nitrogen convertion in Biomass, Highest Concentration and the study variables independent (Ta-feeding time and add urea-MUT). The best results were with (Ta-feeding time=14 days; add urea-MUT=1750 mg). In this conditions we had: Productivity=117,26 mg/L.day Best Gamma-linolenic production condition=23,5 per cent Nitrogen convertion in Biomass=4,9 Biggest Concentration.=1759 mg/L.

Realizou-se um estudo do processo descontínuo alimentado de fermentação, buscando encontrar as melhores condições de produção do ácido graxo gama-linolênico. Empregou-se como fonte de nitrogênio no meio de fermentação a uréia, que foi adicionada de forma exponencialmente crescente e como inóculo usou-se uma cepa de Spirulina platensis. Utilizou-se planejamento experimental e aplicou-se a metodologia de superfície de resposta, o qual permitiu a obtenção de equações que relacionaram as variáveis dependentes, como produtividade em células, conversão de nitrogênio em células, concentração máxima atingida de biomassa, porcentagem relativa de produção de ácido graxo gama-linolênico. com as variáveis independentes tempo de alimentação (Ta) e massa total de uréia adicionada (MUT). Os melhores resultados obtidos foram os seguintes: Px=117,26 mg/L.dia; Yx/n= 4,9; Xmax= 1759 mg/L e porcentagem de ácido graxo γ-linolênico na fração lipídica de 23,5 %, com 14 dias (Ta) e 1750 mg (MUT). / A Study on the fed-batch fermentation process was made seeking for the best Gamma-linolenic production condition. Mineral media was employed with urea as nitrogen source that was add with exponentially increasing feeding rates and Spirulina platensis as inoculum. Experimental design and a Response Surface Methodology were applied. It was composed by an experimental design which allowed a reduced number of the experiments and by the analysis of the results using equations which combined the interest variables dependent as: Productivity, best Gamma-linolenic production condition, Nitrogen convertion in Biomass, Highest Concentration and the study variables independent (Ta-feeding time and add urea-MUT). The best results were with (Ta-feeding time=14 days; add urea-MUT=1750 mg). In this conditions we had: Productivity=117,26 mg/L.day Best Gamma-linolenic production condition=23,5 per cent Nitrogen convertion in Biomass=4,9 Biggest Concentration.=1759 mg/L.

Phosphorus is the main limiting nutrient for plant growth in large areas of the world and the availability of phosphorus to plants and microbes can be strongly affected by soil properties. Even though the phosphorus cycle has been studied extensively, much remains unknown about the key processes governing phosphorus availability in different environments. In this thesis the complex dynamics of soil phosphorus and its availability were studied by relating various phosphorus fractions and soil characteristics to microbial respiration kinetics. The soils used represent a range of aluminium, iron, carbon and total phosphorus content, and were located in four different biomes: subtropical forest, warm temperate forest, boreal forest and tundra. The results showed that NaOH extractable phosphorus, a fraction previously considered to be available to plants only over long time scales, can be accessed by microbes in days or weeks. Microbial phosphorus availability was not related to aluminium or iron content in any of the studied systems, not even in highly weathered soils with high aluminium and iron content. This is in contrast with other studies of soils with high sorption capacity and shows the variability of factors that govern phosphorus availability in different environments. In the boreal forest chronosequence, no difference could be seen with age in total phosphorus content or concentrations of occluded phosphorus forms. However, there were lower concentrations of labile phosphorus forms in older systems, which were correlated with a decrease in microbial respiration. This was most likely related to organic matter quality in the system, and not to geochemical factors. Phosphorus availability was linked to differences in topography (water regime) and vegetation in the tundra ecosystems. The results suggest that the availability of phosphorus, both for microbes and plants, was lower on the meadow vegetation sites compared to the two types of heath vegetation. Many factors are important for phosphorus availability in soils, but these results suggest that microbes can access less available phosphorus if not restricted by carbon, and this may be important in regard to forest management practices as well as effects of environmental change.

Zur Beschreibung von realen Prozessen werden oft Differentialgleichungen herangezogen. Liegen nun Messdaten von diesen Prozessen vor, so sollen auch die Parameter im mathematischen Modell so gewählt werden, dass diese den Messungen entsprechen. Dieser Vortrag zeigt, wie dies in Mathcad mit der Funktion genfit realisiert werden kann.

Submitted by Priscila Nascimento (pnascimento@icict.fiocruz.br) on 2012-11-30T12:31:09Z No. of bitstreams: 1 beatriz-cyranka.pdf: 1720638 bytes, checksum: c436adb348584d5a5a8213fccdef6ea6 (MD5) / Made available in DSpace on 2012-11-30T12:31:09Z (GMT). No. of bitstreams: 1 beatriz-cyranka.pdf: 1720638 bytes, checksum: c436adb348584d5a5a8213fccdef6ea6 (MD5) Previous issue date: 2011 / Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil. / Novas tecnologias, como o sistema isolador para ensaios de esterilidade vieram a contribuir com a incorporação de um melhor desempenho destes ensaios no controle de qualidade das indústrias farmacêuticas.Este trabalho teve como objetivo a verificação do processo de biodescontaminação dentro do sistema isolador de Bio-Manguinhos, utilizado como agente esterilizante o gás de peróxido de hidrogênio. Foram utilizadas como biocarga para redução de contaminação microbiológica três concentrações dos microrganismos, Candida albicansATCC 10231, Clostridium sporogenesATCC 11437 eMicrococcus luteusATCC 9341, impregnadas em discos de filtro de celulose. Os estudos de cinética de crescimento dos microrganismos foram realizados para um melhor entendimento do seumetabolismo, bem como aspectos gerais de crescimento que contribuíram para ressaltar que a Candida albicans inicia sua fase exponencial de crescimento na segunda hora do cultivo e finaliza esta etapa na sexta hora do cultivo, com produção máxima de células viáveis, fato observado também no microrganismo Micrococccus luteus. Para o cultivo doClostridium sporogenes o crescimento foi mais lento com uma curva de crescimento com 60 horas de cultivo total. A produção de maior número de células para o Clostridiumfoi alcançada na vigésima quarta hora de cultivo, assim como a maior produção de esporos. Foi estabelecida, ao longo da curva de crescimento, a relação entre densidade ótica e número de células viáveis, relação essa importante para o estabelecimento das condições do estudo em relação à biocarga empregada de cada microrganismo no momento do desafio no sistema isolador. A capacidade de descontaminação avaliada dentro do sistema isolador com o biocida peróxido de hidrogênio revelou o tempo de exposição ao gás de 10 minutos, como resultado final satisfatório apresentando redução total da carga microbiana com destruição total das células viáveis, assim como as formas esporuladas do Clostridium sporogenes. Desta forma conclui-se que o peróxido de hidrogênioé um biocida de eficácia comprovada, nas variáveis deste estudo e o processode descontaminação no sistema isolador de Bio-Manguinhos é compatível comsua atividade finalística na produção de insumos para a saúde. / New technologies such as isolator system for sterility tests came to help with the incorporation of a better performance of these kinds of tests in quality control of pharmaceutical companies. This study aimed to verify the decontamination process within the system isolator of Bio-Manguinhos, usinghydrogen peroxide gas as a sterilizing agent. Three concentrations of microorganisms were used as bioburden for the microbiological contamination reduction, Candida albicansATCC 10231, Clostridium sporogenesATCC 11437 and Micrococcus luteusATCC 9341, impregnated in cellulose filter discs. Studies of growth kinetics of microorganisms were carried out to a better understanding of theirmetabolism, as well as general aspects of growth that contributed to emphasize that the Candida albicansbegins its exponential growth phase in the second hour of cultivation and this step ends at the sixth hour cultivation, with maximum yield of viable cells, this was also observed in the Micrococcus luteusmicroorganism. For the Clostridium sporogenescultivation, growth was slower with a 60 hours growth curve of total culture. The production of more cells for Clostridiumwas achieved in the twenty-fourth hour of cultivation, as well as the maximum spores production. It was established along the growth curve the relationship between optical density and numberof viable cells, this relationship was important to establish the conditions of the study related to the bioburden of each microorganism used to challenge the isolator system. The decontamination capacity evaluated within the isolator system with the hydrogen peroxide biocide showed that the gas exposure time of 10 minutes wassatisfactory demonstrating total reduction of the microbial load with total destruction of viable cells, as well as the sporulated forms of Clostridium sporogenes. Thus it is concluded that hydrogen peroxide is a proved effective biocide, in the variables of this study and decontamination process in the Bio-Manguinhos insulator system is compatible with its main activity in the production of health supplies.

Ces travaux s'appuient sur un contexte de santé lié à la dégradation de la qualité de l'air intérieur induite par la présence de micro-organismes. Dans les environnements intérieurs humides, les matériaux de construction sont des cibles de contamination et de prolifération microbienne importantes. La photocatalyse est un procédé de dépollution qui présente une action contre une large gamme de polluants organiques (aqueux, gazeux ou biologiques). Son principe repose sur l'excitation d'un photocatalyseur par une irradiation lumineuse, généralement située dans les UV, qui va permettre de dégrader les polluants environnants par une succession de réactions d'oxydo-réduction. Le photocatalyseur le plus courant est le dioxyde de titane (TiO2). Outre la production d'espèces oxydo-réductrices agressives, le TiO2 illuminé présente également un caractère super-hydrophile qui lui confère un caractère autonettoyant intéressant. Une autre solution est envisagée pour lutter contre la prolifération microbienne sur matériaux de construction : l'utilisation de molécule bio-sourcées d'ester de glycérol aux propriétés naturellement antimicrobienne. L'objectif de cette étude est de développer des revêtements pour matériaux de construction intérieurs et d'étudier leur propriété de résistance à la prolifération microbienne et les modes d'action de ces dispositifs passifs formulés soit à base de particules de TiO2, soit à base d'ester de gycérol. Dans un premier temps, un travail de développement et de mise en œuvre de dispositifs expérimentaux a été nécessaire afin d'adapter des méthodes d'évaluation microbiologiques sur ces matériaux particuliers (particules, lasures, matériaux cimentaires). Ainsi, plusieurs essais ont été adaptés afin d'évaluer les performances antimicrobiennes en terme (i) d'activité antibactérienne, (ii) d'effet bactéricide, (iii) de résistance à la formation de biofilm et (iv) de résistance à la prolifération par contamination " naturelle ". Les premiers essais visent à mettre en évidence l'impact des facteurs méthodologiques sur l'activité antibactérienne des particules de TiO2 utilisées seules comme agent désinfectant. Dans un deuxième temps, des lasures photocatalytiques sont formulées sur la base de travaux antérieurs ayant montrés de bonnes efficacités de dépollution de l'air contre les NOx, NO et différents COV. Une fois les paramètres d'influence de l'activité du TiO2 mis en exergue, les lasures ont été testées dans des conditions optimales. Le développement d'un essai de résistance à la prolifération de biofilm montre l'importance de coupler différentes méthodes d'évaluation microbiologique (dénombrement des UFC et observation au microscope à épifluorescence). La dernière partie de cette étude explore le potentiel antimicrobien de la molécule d'ester de glycérol, sous-produit de la synthèse de biocarburant. La molécule montre des propriétés antibactériennes et bactéricides puissantes en quelques minutes de contact seulement. Elle présente également une protection très efficace contre la prolifération microbienne une fois recouverte sur matériaux de construction (placo-plâtre). Ces performances remarquables encouragent la poursuite des études sur cette molécule. / This work is included in a health-related context: the degradation of the indoor air quality induced by the presence of microorganisms. In damp environments, indoor building materials are among the main proliferation substrates for microorganisms. Photocatalysis is a decontamination process which is active against a wide range of organic pollutants (aqueous, gaseous or biological). The principle is based on the excitation of a photocatalyst by light irradiation, usually located in the UV-range, which leads to the degradation or mineralization of surrounding pollutants through a series of oxidative reactions. The most common photocatalyst is titanium dioxide (TiO2). In addition to produce aggressive redox species, the illuminated TiO2 also shows super-hydrophilicity, which has an impact on the first step of microbial biofilm formation: the adhesion of microorganisms. Another technology to protect indoor building materials is explored: the use of glycerol esters, which are bio-based molecules with inherent antimicrobial properties. The main objective of this study is to develop semi-transparent coatings for indoor building materials and to study the resistance to microbial growth conferred by these passive devices, which are formulated using TiO2 nanoparticles or glycerol esters. Initially, the development and implementation of experimental devices has been necessary to adapt microbiological evaluation methods to these particular materials (nanoparticles, surface coatings, cementitious materials). Thus, several tests have been carried out in order to assess the antimicrobial performances in terms of (i) antibacterial activity, (ii) bactericidal effect, (iii) resistance to biofilm formation and (iv) resistance to proliferation by "natural" contamination. First tests aimed to underline the influence of methodological parameters on the efficiency of TiO2 particles used alone as antimicrobial agent. Then, semi-transparent coatings were formulated on the basis of previous works which have shown good efficiencies on the depollution of ambient air from NOx, NO and VOC. Once the parameters that influence TiO2 activity were identified, coatings were tested in optimum conditions. The evaluation of the resistance to biofilm formation shows the importance of overlapping different microbiological evaluation methods (e.g. CFU counting and epifluorescence observations). The last part was an exploratory work on the antimicrobial properties of a glycerol ester molecule, by-product from the synthesis of biofuels. The molecule shows potent antibacterial and bactericidal properties, several log of inactivation within only few minutes of contact. It also provides very effective protection against microbial growth once covered on building materials (plasterboard). These remarkable performances encourage further studies on this molecule.

Made available in DSpace on 2015-03-26T13:55:09Z (GMT). No. of bitstreams: 1 texto completo.pdf: 813429 bytes, checksum: 8b1f7f1d9b802a3b69cd3247045c7ac3 (MD5) Previous issue date: 2012-02-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study evaluated the effects of replacing soybean meal (SBM) by slow-release urea (SRU) in beef cattle diets containing two concentrate levels on intake, partial and total digestibility, nutrients flow, ruminal parameters, microbial protein production, protein efficiency and nitrogen balance in finishing beef steers. Eight crossbred steers were used (average BW of 418 kg) fitted with ruminal and abomasal cannulae. The animals were distributed in two groups of four steers, each group received one of the two levels of the concentrate (40 or 80% of the total DM). The experiment was designed as a Latin square in 2 x 4 factorial scheme, with two energy levels in the diet, and within each square, four levels of replacement of soybean meal protein by slow-release urea: 0, 33, 66 and 100 %. The interaction of concentrate level x SRU levels was not significant (P>0.05) for any variables measure. Thus, the effects of concentrate level and SRU inclusion will be discussed independently. Dry matter (DM) intake, as well as organic matter (OM) intake and crude protein (CP) intake decreased linearly (P<0,05) as SBM was replaced by SRU. For each percentage unit of SRU added to the diet there was a reduction of 5.80, 5.17 and 0.58 g in DMI, OMI and CPI. The intake of non-fiber carbohydrates (NFC) was higher (P <0.05) in the diet containing 80% concentrate when compared to the 40 % concentrate diet, whereas neutral detergent fiber (NDF) intake was greater (P <0.05) on the low concentrate diet when compared to the high concentrate diet. Ruminal digestibility coefficient of OM tented to be greater (P <0.074) for the 40 % concentrate diet, whereas all other ruminal, intestinal and total digestibility parameters evaluated were not statistically different (P> 0.10) among treatments. DM and OM passage rates (kp) were greater (P<0.05) on the 80% concentrate diet than on the 40 % concentrate diet, with averages of 4.37 and 4.35%. Replacing SBM by SRU up to 100 % on the diet did not lead to any modification (P>0.05) on DM, OM, CP and NDF passage rate. Digestion rates (kd) were also not affected by any of the treatments (P>0.05). The average kd for DM, OM, CP and NDF were 3.82, 4.21, 1.13 and 2.20%. A cubic effect (P <0.10) of SBM replacement by SRU on ruminal ammonia (N-NH3) concentration in relation to the hours of collection was detected. A quadratic effect was observed (P <0.10) by replacing SBM with SRU on pH and on the excretion of urea N in urine, expressed both as g/day or mg/kg of BW. The animals had higher (P <0.10) total N excretion in urine, in g/day, as the slow-release urea levels were increased in the diet, but no significant effects were observed (P>0.10) for total N in urine in mg/kgBW (average of 148.56 mg/kgBW) and in the concentrations of urea N in the serum (average value of 11.53 mg/dL). Nitrogen intake, N excreted in the feces, nitrogen balance and efficiency of nitrogen use decreased linearly (P <0.10) as the levels of SRU increased in the diet, whereas the amount of nitrogen compounds excreted in urine increased linearly (P <0.10) as SRU replaced SBM in the diet. The production of microbial nitrogen and microbial efficiency were not significantly affected by the experimental treatment (P> 0.10) obtaining average values of 152.02 g/day and 120.18 g/kg TDN, respectively. / Objetivou-se avaliar os efeitos da substituição do farelo de soja por uréia de liberação lenta (0, 33, 66 e 100%) em dietas com dois níveis de concentrado (40 e 80%) sobre o consumo, digestibilidade parcial e total, fluxo de nutrientes, parâmetros ruminais, produção de proteína microbiana, eficiência proteica e balanço de nitrogênio em novilhos de corte em terminação. Foram utilizados oito novilhos de corte cruzados (Bos taurus x Bos indicus), com peso corporal médio (PC) de 418 ± 40 kg e fistulados no rúmen e abomaso, que foram divididos em dois grupos de quatro, em que cada grupo recebeu um nível de concentrado. A alimentação basal foi composta de silagem de milho e o concentrado foi formulado à base de milho moído fino, farelo de soja, uréia de liberação lenta e mistura mineral. O experimento foi avaliado segundo delineamento em quadrado latino, com agrupamento de dois quadrados simultâneos. Dentro de cada quadrado, foram implementados quatro tratamentos, relativos à substituição da proteína do farelo de soja por uréia de liberação lenta (256 % de PB): 0, 33, 66 e 100 %. Verificou-se efeito linear decrescente (P<0,10) do nível de inclusão da uréia de liberação lenta sobre o consumo de matéria seca (MS), de proteína bruta (PB), de matéria orgânica (MO), de MS em relação ao peso corporal e dos teores de concentrado sobre o consumo de fibra em detergente neutro corrigida para cinzas e proteína (FDNcp), carboidratos não fibrosos (CNF) e FDNcp em relação ao peso corporal. Somente o coeficiente de digestibilidade ruminal da MO foi influenciado (P<0,10) pelo nível de concentrado na dieta, em que o menor nível de concentrado apresentou o maior valor para esta variável. Para todos os outros parâmetros de digestibilidade não houve diferença estatística (P>0,10) entre os tratamentos avaliados. As dietas com 80% de concentrado promoveram maior taxa de passagem (kp) da matéria seca e da matéria orgânica, não havendo diferenças estatísticas (P>0,10) para as kps da PB e da FDNcp. As variáveis relacionadas à taxa de digestão não foram influenciadas (P>0,10) por nenhum dos tratamentos avaliados. Verificou-se efeito cúbico (P<0,10) do nível de substituição do farelo de soja por uréia de liberação lenta sobre as concentrações de N-NH3 e quadrático para o pH em relação às horas de coleta para cada nível de concentrado testado Encontrou-se efeito quadrático (P<0,10) para as excreções de nitrogênio uréico na urina em gramas por dia e em miligramas por quilo de peso corporal. Os animais apresentaram maiores excreções de nitrogênio total na urina em gramas por dia conforme foram aumentados os níveis de uréia de liberação lenta, porém não foram encontrados efeitos significativos para o nitrogênio total na urina em mg/kg de PC, nas concentrações de nitrogênio uréico no soro e na relação NUU/NU. Os compostos nitrogenados ingeridos, excretados nas fezes, o balanço dos compostos nitrogenados e a eficiência de uso do nitrogênio decresceram linearmente (P<0.10) conforme se aumentava os níveis de ULL nas dietas, ao passo que os compostos nitrogenados excretados na urina cresceram linearmente (P<0.10) com o aumento de ULL. Já o nitrogênio microbiano e a eficiência microbiana não foram afetados significativamente (P>0,10) pelos tratamentos avaliados. A ULL diminui o consumo de MS, MO e PB de bovinos em terminação, mantendo, entretanto, o consumo de nutrientes digestíveis totais inalterado. O uso de ULL em substituição ao farelo de soja em dietas para bovinos em terminação não altera a eficiência e a produção de proteína microbiana, a digestibilidade e nem as taxas de digestão e passagem da MS, MO, PB e FDNcp. A substituição do arelo de soja por uréia de liberação lenta permite melhor recuperação do pH ruminal após atingir o valores mínimos em dietas com alto concentrado quando comparada ao farelo de soja.

Aktivt slam är en biologisk process som används i stor utsträckning för rening av avloppsvatten, där bakterier som odlas under luftiga förhållanden avlägsnar organiska ämnen och näringsämnen från förorenat vatten. För industriell verksamhet modelleras denna process för att representera och förutsäga biologiska fenomen. De modeller som används för närvarande (främst modellen för aktiverat slam) ger acceptabla resultat men har vissa begränsningar, t.ex. lång kalibreringstid och beroende av många modellparametrar som är svåra att förutsäga. Under detta projekt som genomfördes på företaget SUEZ bedömdes det om sådana modeller kunde förbättras genom att lägga till ett termokemiskt perspektiv i deras arkitektur, antingen genom att minska antalet parametrar eller genom att teoretiskt förutsäga deras värden. Det termokemiska bidraget bedömdes på två olika nivåer: de stökiometriska förhållanden som beskriver systemet och de biologiska processernas kinetik. På grundval av en omfattande litteraturstudie valdes två metoder ut för att föra in termodynamiska överväganden i dessa delar av modellen. Termodynamiska beräkningar som krävs för att förbättra specifikationen av de förorenande molekyler som finns i vattenmatrisen. Arkitekturen i den ursprungliga modellen för aktiverat slam ändrades för att möjliggöra en differentiering mellan substrat av olika slag. När det gäller den stökiometriska studien visade det sig att den dynamiska förutsättningen av termodynamiska begränsningar för reaktionerna inte var av något större intresse med tanke på de stora mängder energi som finns tillgängliga. Användningen av olika organiska substrat gav dock intressanta resultat, eftersom det gjorde det möjligt att modellera de biologiska beteendena på ett mer detaljerat sätt och att förutsäga tillväxten av specifika typer av mikroorganismer. Den kinetiska studien gav måttliga resultat, med adekvata simuleringar men utan någon ny större förbättring av modellen. Den mest lovande utvecklingen av denna studie tycks vara en modell som omfattar en mängd olika organiska substrat med motsvarande biomassapopulationer, för att kunna rikta in sig på den specifika förekomsten av intressanta bakterier. / Activated sludge is a biological process that is widely used for wastewater treatment, where bacteria grown in aerated conditions remove organic substances and nutrients from polluted waters. For industrial operations, this process is modeled to represent and predict biological phenomena. Currently used models (mainly, the Activated Sludge Model) give acceptable results but suffer from some limitations such as long calibration time and dependency on an many model parameters that are difficult to predict. During this project conducted at the company SUEZ, it was assessed whether such models could be improved by adding a thermochemical perspective to their architecture, either by reducing the number of parameters involved or by predicting theoretically their values. The contribution of thermochemistry was assessed on two different levels: the stoichiometric relations that describe the system, and the kinetics of the biological processes. Based on an extensive literature study, two methods were selected to inject thermodynamical considerations to these parts of the model. Thermodynamic calculations required to improve the specification of the pollutant molecules present in the water matrix. The architecture of the initial Activated Sludge Model was modified to allow for a differentiation between substrates of different nature. Regarding the stoichiometric study, it appeared that the dynamic prevision of thermodynamical constraints over the reactions did not show a major interest considering the high amounts of energy available. However, the use of different organic substrates gave interesting results because it allowed to model more finely the biological behaviors and to predict the growth of specific types of microorganisms. The kinetic study gave mitigated results, with adequate simulations but no new major improvement to the model. The most promising development of this study appears to be a model involving a multiplicity of organic substrates, with corresponding biomass populations, to target the specific occurrence of bacteria of interest.

La sécrétion de lipase, la lipolyse extracellulaire et l’absorption des acides gras (AGL) ont été étudiés chez Yarrowia lipolytica (YL) en présence d’huile d’olive et/ou de sucrose. Des mesures d’activité lipase et d’immuno-révélation ont montré que l’activité lipase présente dans le milieu de culture provenait principalement de la lipase YLLIP2. L’huile d’olive induit la production de lipase qui est principalement associée aux cellules pendant les premières heures de cultures. YLLIP2 est ensuite libérée dans le milieu de culture avant d’être totalement dégradée par les protéases. Les triglycérides (TG) sont dégradés alors que la lipase est encore attachée aux cellules. Les produits de lipolyse présents dans le milieu de culture et à l’intérieur des cellules ont été quantifiés par chromatographie TLC-FID et GC. Les niveaux intracellulaires d’AGL et de TG augmentent transitoirement et dépendent de la source de carbone utilisée. Une accumulation maximum de 37,8 % w/w de lipides est observée avec l’huile d’olive seule. Cette étude montre que la levure YL est un modèle intéressant pour étudier la lipolyse extracellulaire et l’absorption des acides gras par les cellules / Lipase secretion, extracellular lipolysis and fatty acid (FFA) uptake were quantified in Yarrowia lipolytica (YL) grown in the presence of olive oil and/or sucrose. Lipase assays and western blot analysis indicated that the lipase activity measured in YL cultures mainly resulted from YLLIP2 lipase. Lipase production was triggered by olive oil and YLLIP2 remained associated with the yeast cells during the first hours of culture. It was then released in the culture medium before it was totally degraded by the alkaline protease. Olive oil triglycerides (TG) were degraded when the lipase was still attached to the cell wall. The fate of lipolysis products in the culture medium and inside the yeast cell were investigated by quantitative TLC-FID and GC analysis. Intracellular levels of FFA and TG increased transiently and were dependent on the carbon sources. A maximum fat storage of 37.8% w/w was observed with olive oil alone. The present study shows that yeasts are interesting models for studying extracellular lipolysis and fat uptake by the cell

Novos sistemas biocompatíveis para aplicações biotecnológicas são relevantes não só do ponto de vista tecnológico, mas também para avanços científicos. A presente dissertação visou à criação e caracterização de filmes finos (espessura de até 100 nm) ou micrométricos (espessura de 50 &#181;m a 100 &#181;m) formados por goma xantana (GXT), poli(cloreto de dimetil dialil amônio), (PDDA), albumina bovina sérica (BSA), e nanopartículas de magnetita (NPM). A incorporação de amoxicilina (Amox), um antibiótico amplamente utilizado contra muitas infecções, nos filmes foi realizada durante a formação dos sistemas. Os filmes finos foram caracterizados através de elipsometria e microscopia de força atômica. Em solução, as interações favoráveis entre o Amox e a BSA foram evidenciadas por mudanças substanciais na estrutura secundária da BSA, como revelado pelo dicroísmo circular. Os filmes micrométricos (patches) de GXT e GXT/NPM/BSA foram imersos em Amox 2 g/L, levando a incorporação de 10 ± 3 e 17 ± 4 &#181;g/cm³ de Amox, respectivamente. Sua caracterização compreendeu espectroscopia vibracional no infravermelho por transformada de Fourier no modo de refletância total atenuada (FTIR-ATR), microscopia eletrônica de varredura (MEV), medidas de sorção e espectrometria de emissão óptica com plasma indutivamente acoplado (ICP-OES). A incorporação de 0,2% em massa de Fe3O4 nos \"patches\" e sua exposição a um campo magnético externo, viabilizou a liberação total in vitro de Amox, em pH 5,5 e em 0,02 mol/L de NaCl, seguindo o comportamento quasi-Fickiano. A difusão da Amox dos \"patches\" de GXT/NPM/BSA em ágar contendo Staphylococcus aureus e Escherichia coli, levou halos de inibição consideráveis. A inibição do crescimento de E. coli foi particularmente eficiente sob efeito de campo magnético externo. / New biocompatible systems for biotech applications are relevant not only from a technological point of view, but also for scientific advances. The present project aimed at the creation and characterization of thin films (thickness up to 100 nm) or micrometric patches (thickness of 50 &#181;m to 100 &#181;m) formed by the deposition of xanthan gum (GXT), poly (dimethyl diallyl ammonium chloride) (PDDA), bovine serum albumin (BSA), and magnetite nanoparticles (NPM). The incorporation of amoxicillin (Amox), a widely used antibiotic against many infections, in the films was performed during the formation of the systems. Thin films were characterized by means of ellipsometry and atomic force microscopy. In solution, favorable interactions between Amox and BSA were evidenced by substantial changes in the secondary structure of BSA, as revealed by circular dichroism spectra. Patches of GXT and GXT/NPM/BSA were immersed into Amox solution at 2 g/L, leading to the Amox incorporation of 10 ± 3 and 17 ± 4 &#181;g/cm3, respectively. The patches characterization included Fourier Transform Infrared vibration spectroscopy in the attenuated total reflectance mode (FTIR-ATR), scanning electron microscopy (MEV), sorption measurements and inductively coupled plasma optical emission spectrometry (ICP-OES). The incorporation of 0.2 wt% of Fe3O4 in the patches and their exposure to an external magnetic field, allowed the total in vitro release of Amox, at pH 5.5 and 0.02 mol/L NaCl, following the behavior quasi-Fickian. Amox diffusion from GXT/NPM/BSA patches in agar containing Staphylococcus aureus and Escherichia coli led to a considerable zone of inhibition. Inhibition of E. coli growth was particularly efficient under the effect of external magnetic field.