P-aminobenzoic acid (PABA) is a polymorphic compound with two known polymorphs - alpha with a needle morphology and β with a rhombic morphology. It is an enantiotropic compound with a transition temperature at 13.8oC, where alpha is more thermodynamically stable above transition temperature and β is more thermodynamically stable below. At the beginning of this project, crash-cooling crystallisation experiments were conducted to determine the effect of solvent, temperature and supersaturation on the nucleating polymorphs of PABA. Three solvents were tested (water, ethanol and isopropyl alcohol) over a range of supersaturations and temperatures. The results suggested that polymorph appearance of PABA was heavily influenced by kinetics, as opposed to thermodynamics of the system, disagreeing with Ostwald's rule of stages. The project then focussed on the ability of tailor-made additives to select the crystallising polymorph of PABA from supersaturated solutions of PABA in isopropyl alcohol. Crash-cooling crystallisation experiments were performed using two additives: 4-amino-3-nitrobenzoic acid, and 4-amino-3-methoxybenzoic acid. Results showed that alpha PABA crystallised below a critical concentration of either additive, and above that critical concentration, β PABA would crystallise. To determine whether the additives were affecting the nucleation and/or growth kinetics of alpha PABA and β PABA, a series of nucleation and growth experiments were conducted using a Crystal16 multiple stirred reactor and a crystal growth cell respectively. The results showed that both additives greatly reduced the attachment frequency of growth units to alpha PABA nuclei, and inhibited the growth rate of alpha PABA seed crystals. Nucleation data could not be obtained for β PABA, but in terms of crystal growth, both additives did not affect growth rate of β PABA to a noticeable degree. Gravimetric and HPLC experiments were also employed to measure the solubility effects of both additives on PABA in isopropyl alcohol. Results showed that both additives did not appear to affect PABA's solubility to a noticeable degree.

Two different aluminium alloys, AA6111 (Al-Mg-Si) and AA7055 (Al-Mg-Zn), were chosen as the aluminium alloys to be welded with DC04, and two welding methods (USW and FSSW) were selected to prepare the welds. Selected pre-welded joints were then annealed at T=400 - 570oC for different times. Kinetics growth data was collected from the microstructure results, and the growth behaviour of the IMC layer was found to fit the parabolic growth law. A grain growth model was built to predict the grain size as a function of annealing time. A double-IMC phase diffusion model was applied, together with grain growth model, to predict the thickness of each phase as a function of annealing time in the diffusion process during heat treatment. In both material combinations and with both welding processes a similar sequence of IMC phase formation was observed during the solid state welding. η-Fe2Al5 was found to be the first IMC phase to nucleate. The IMC islands then spread to form a continuous layer in both material combinations. With longer welding times a second IMC phase, θ-FeAl3, was seen to develop on the aluminium side of the joints. Higher fracture energy was received in the DC04-AA6111 joints than in the DC04-AA7055 joints. Two reasons were claimed according to the microstructure in the two joints. The thicker IMC layers were observed in the DC04-AA7055 joints either before or after heat treatment, due to the faster growth rate of the θ phase. In addition, pores were left in the aluminium side near the interface as a result of the low melting point of AA7055.The modelling results for both the diffusion model and grain growth model fitted very well with the data from the static heat treatment. Grain growth occurred in both phases in the heat treatment significantly, and was found to affect the calculated activation energy by the grain boundary diffusion. At lower temperatures in the phases with a smaller grain size, the grain boundary diffusion had a more significant influence on the growth rate of the IMC phases. The activation energies for the grain boundary diffusion and lattice diffusion were calculated as 240 kJ/mol and 120 kJ/mol for the η phase, and 220 kJ/mol and 110 kJ/mol for the θ phase, respectively. The model was invalid for the growth of the discontinuous IMC layers in USW process. The diffusion model only worked for 1-Dimensional growth of a continuous layer, which was the growth behaviour of the IMC layer during heat treatment. However, due to the highly transient conditions in USW process, the IMC phases were not continuous and uniform even after a welding time of 2 seconds. Therefore, the growth of the island shaped IMC particles in USW was difficult to be predicted, unless the nucleation stage was taken into consideration.

No período de 1999 a 2002, uma linhagem de Salmonella Enteritidis esteve envolvida em mais de 90% das salmoneloses ocorridas no RS. Este trabalho teve por objetivo avaliar a cinética de crescimento, a resistência ácida e a resistência térmica dessa linhagem e compará-la com S. Typhimurium e S. Bredeney não envolvidas em surtos alimentares, porém isoladas na mesma região. Em uma primeira etapa, a cinética de crescimento foi avaliada semeando-se cada sorovar em caldo nutriente (CN) e em salada de batata com maionese caseira (SMC), os quais foram mantidos a 30°C e 9,5°C. Em CN, a cinética de crescimento a 30°C foi semelhante para todos os sorovares, porém, em SMC a S. Enteritidis apresentou maior quantidade de células nas primeiras 6 horas de crescimento, sendo que somente depois de 12 horas todos os sorovares atingiram quantidades semelhantes de células. Em CN e em SMC, na temperatura de 9,5°C, não foi detectado crescimento de nenhum dos sorovares de Salmonella durante as primeiras 24 horas, sugerindo que essa temperatura foi suficiente para controlar a multiplicação desses microrganismos. Em uma segunda etapa, avaliou-se a resistência ácida e térmica dos diferentes sorovares de Salmonella. Para tanto, os três sorovares foram inoculados separadamente em CN e CN enriquecido com 1% de glicose (CNG), este último utilizado para produção de culturas ácido-adaptadas. Em seguida, os microrganismos foram submetidos a diferentes pH (3,5; 4,0 e 4,5) e temperaturas (52, 56 e 60ºC). Os resultados demonstraram que a S. Bredeney apresentou maior resistência para os pH 3,5 e 4,0, porém a S. Enteritidis demonstrou maior capacidade de adaptação ácida do que S. Typhimurium e S. Bredeney. Em pH 4,5 todos os sorovares, tanto não adaptados quanto ácido-adaptados, mantiveram a mesma quantidade de células viáveis durante 300 minutos. Quando expostas a 52ºC, S. Bredeney apresentou maior resistência, entretanto somente a S. Enteritidis foi protegida com a adaptação ácida. Para 56 e 60ºC, a S. Enteritidis, não adaptada e ácido-adaptada, apresentou maior resistência. A análise por SDS-PAGE demonstrou diferenças no perfil protéico de células não adaptadas e ácidoadaptadas para todos os sorovares testados. Com base nestes resultados, a capacidade de multiplicação mais rápida nas primeiras horas de cultivo em SMC, a maior capacidade de adaptação ácida e a maior resistência térmica demonstradas pela S. Enteritidis podem estar relacionadas ao freqüente envolvimento desse sorovar nas salmoneloses do RS. / During the period of 1999 to 2002, a strain of Salmonella Enteritidis was involved in more than 90 % of foodborne salmonellosis occurred in Rio Grande do Sul (RS) State. This work aimed to evaluate the growth kinetics, and the acid and the thermal resistance of this strain, comparing with S. Typhimurium and S. Bredeney, which were not involved in foodborne outbreaks, but were isolated in the same region. In the first stage of this study, the growth kinetics was assessed. Each serovar was inoculated separately in nutrient broth (CN) and in potato salad prepared with homemade mayonnaise (SMC), and then incubated at 30 ºC and 9.5 ºC. In CN, at 30 ºC, similar growing characteristics were found for all serovars, however in SMC S. Enteritidis demonstrated higher counts at the first 6 hours. Only after 12 hours of incubation, all serovars reached similar counts. In CN and in SMC, at 9.5 ºC, during the first 24 hours, there was no detectable growth of any Salmonella serovar, suggesting that such temperature was adequate to control the multiplication of tested Salmonella serovars. In the second stage of the study, the acid and the thermal resistances of Salmonella serovars were evaluated. The three serovars were cultivated separately in Nutrient Broth and Nutrient Broth supplemented with 1 % glucose (NBG). The latter medium was used to induce acid-adapted cells. Then, the three serovars were exposed to different pH (3.5, 4.0, and 4.5) and temperatures (52, 56, and 60 ºC). Results indicated that S. Bredeney presented higher resistance to pH 3.5 and 4.0, but S. Enteritidis presented a better capacity of acid adaptation than S. Typhimurium and S. Bredeney. At pH 4.5, all serovars demonstrated a similar behavior, remaining at same levels of viable cells until 300 minutes. At 52 ºC, S. Bredeney presented greater survivor rates, however acid adaptation protected only S. Enteritidis. At 56 ºC and 60 ºC, non-adapted and acidadapted S. Enteritidis were more thermally resistant than other serovars tested. SDS-PAGE analysis demonstrated differences in protein profile of non-adapted and acid-adapted cells of all serovars. The capacity of rapid multiplication in the first hours of cultivation, the greater acid adaptation and thermal resistance presented by S. Enteritidis, may be related to the frequent involvement of this strain in salmonellosis cases in the RS.

No período de 1999 a 2002, uma linhagem de Salmonella Enteritidis esteve envolvida em mais de 90% das salmoneloses ocorridas no RS. Este trabalho teve por objetivo avaliar a cinética de crescimento, a resistência ácida e a resistência térmica dessa linhagem e compará-la com S. Typhimurium e S. Bredeney não envolvidas em surtos alimentares, porém isoladas na mesma região. Em uma primeira etapa, a cinética de crescimento foi avaliada semeando-se cada sorovar em caldo nutriente (CN) e em salada de batata com maionese caseira (SMC), os quais foram mantidos a 30°C e 9,5°C. Em CN, a cinética de crescimento a 30°C foi semelhante para todos os sorovares, porém, em SMC a S. Enteritidis apresentou maior quantidade de células nas primeiras 6 horas de crescimento, sendo que somente depois de 12 horas todos os sorovares atingiram quantidades semelhantes de células. Em CN e em SMC, na temperatura de 9,5°C, não foi detectado crescimento de nenhum dos sorovares de Salmonella durante as primeiras 24 horas, sugerindo que essa temperatura foi suficiente para controlar a multiplicação desses microrganismos. Em uma segunda etapa, avaliou-se a resistência ácida e térmica dos diferentes sorovares de Salmonella. Para tanto, os três sorovares foram inoculados separadamente em CN e CN enriquecido com 1% de glicose (CNG), este último utilizado para produção de culturas ácido-adaptadas. Em seguida, os microrganismos foram submetidos a diferentes pH (3,5; 4,0 e 4,5) e temperaturas (52, 56 e 60ºC). Os resultados demonstraram que a S. Bredeney apresentou maior resistência para os pH 3,5 e 4,0, porém a S. Enteritidis demonstrou maior capacidade de adaptação ácida do que S. Typhimurium e S. Bredeney. Em pH 4,5 todos os sorovares, tanto não adaptados quanto ácido-adaptados, mantiveram a mesma quantidade de células viáveis durante 300 minutos. Quando expostas a 52ºC, S. Bredeney apresentou maior resistência, entretanto somente a S. Enteritidis foi protegida com a adaptação ácida. Para 56 e 60ºC, a S. Enteritidis, não adaptada e ácido-adaptada, apresentou maior resistência. A análise por SDS-PAGE demonstrou diferenças no perfil protéico de células não adaptadas e ácidoadaptadas para todos os sorovares testados. Com base nestes resultados, a capacidade de multiplicação mais rápida nas primeiras horas de cultivo em SMC, a maior capacidade de adaptação ácida e a maior resistência térmica demonstradas pela S. Enteritidis podem estar relacionadas ao freqüente envolvimento desse sorovar nas salmoneloses do RS. / During the period of 1999 to 2002, a strain of Salmonella Enteritidis was involved in more than 90 % of foodborne salmonellosis occurred in Rio Grande do Sul (RS) State. This work aimed to evaluate the growth kinetics, and the acid and the thermal resistance of this strain, comparing with S. Typhimurium and S. Bredeney, which were not involved in foodborne outbreaks, but were isolated in the same region. In the first stage of this study, the growth kinetics was assessed. Each serovar was inoculated separately in nutrient broth (CN) and in potato salad prepared with homemade mayonnaise (SMC), and then incubated at 30 ºC and 9.5 ºC. In CN, at 30 ºC, similar growing characteristics were found for all serovars, however in SMC S. Enteritidis demonstrated higher counts at the first 6 hours. Only after 12 hours of incubation, all serovars reached similar counts. In CN and in SMC, at 9.5 ºC, during the first 24 hours, there was no detectable growth of any Salmonella serovar, suggesting that such temperature was adequate to control the multiplication of tested Salmonella serovars. In the second stage of the study, the acid and the thermal resistances of Salmonella serovars were evaluated. The three serovars were cultivated separately in Nutrient Broth and Nutrient Broth supplemented with 1 % glucose (NBG). The latter medium was used to induce acid-adapted cells. Then, the three serovars were exposed to different pH (3.5, 4.0, and 4.5) and temperatures (52, 56, and 60 ºC). Results indicated that S. Bredeney presented higher resistance to pH 3.5 and 4.0, but S. Enteritidis presented a better capacity of acid adaptation than S. Typhimurium and S. Bredeney. At pH 4.5, all serovars demonstrated a similar behavior, remaining at same levels of viable cells until 300 minutes. At 52 ºC, S. Bredeney presented greater survivor rates, however acid adaptation protected only S. Enteritidis. At 56 ºC and 60 ºC, non-adapted and acidadapted S. Enteritidis were more thermally resistant than other serovars tested. SDS-PAGE analysis demonstrated differences in protein profile of non-adapted and acid-adapted cells of all serovars. The capacity of rapid multiplication in the first hours of cultivation, the greater acid adaptation and thermal resistance presented by S. Enteritidis, may be related to the frequent involvement of this strain in salmonellosis cases in the RS.

Submitted by LUCAS PORTILHO DA CUNHA null (lucasportilhodacunha@gmail.com) on 2016-04-21T16:42:26Z No. of bitstreams: 1 DISSERTAÇÃO_LUCAS PORTILHO DA CUNHA VERSÃO FINAL.pdf: 2114741 bytes, checksum: 6122b7305d3f81a525845b852fac3200 (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-05-02T16:39:31Z (GMT) No. of bitstreams: 1 cunha_lp_me_sjrp.pdf: 2114741 bytes, checksum: 6122b7305d3f81a525845b852fac3200 (MD5) / Made available in DSpace on 2016-05-02T16:39:31Z (GMT). No. of bitstreams: 1 cunha_lp_me_sjrp.pdf: 2114741 bytes, checksum: 6122b7305d3f81a525845b852fac3200 (MD5) Previous issue date: 2016-03-28 / O presente trabalho teve como objetivo principal o desenvolvimento de um biorreator de bandeja capaz de produzir esporos do fungo entomopatogênico Metarhizium anisopliae. Para esse fim, foi utilizado um biorreator de bandeja para a produção de esporos, que por sua vez foi avaliada em função da carga de substrato empregada. As variações de temperatura no biorreator foram simuladas e observadas experimentalmente. Para o desenvolvimento do biorreator, inicialmente foram realizados ensaios em embalagens plásticas contendo 10g e 500g de substrato, de modo a avaliar como o aumento da carga de meio fermentativo poderia influenciar na produção de esporos. Nestes ensaios, foram medidas as temperaturas no centro geométrico das embalagens e a elevação de temperatura foi relacionada à produção de esporos. Realizou – se uma análise de área superficial do arroz tipo 1 e da quirera de arroz para avaliar a interferência desta propriedade na produção de esporos. A partir dos resultados em embalagens plásticas foram realizados experimentos em um biorreator de 40 cm de comprimento, 29 cm de largura por 12 cm de altura, escoando ar sobre as partículas no sentido da maior dimensão. Foram empregadas cargas de 1, 2 e 3kg de arroz tipo 1, correspondente à altura de leito de 2, 4 e 6 cm, respectivamente, e 1 kg de quirera, correspondendo a 2 cm de altura. A temperatura nas posições de entrada e saída de ar e no meio geométrico do meio de cultivo foram medidas ao longo dos ensaios. A fermentação nas embalagens plásticas e no biorreator foram realizadas a 28oC. Para estimar a geração de calor metabólico do meio de cultivo, foram coletados dados de consumo de O2 e liberação de CO2 durante o período de incubação. Para tal, foi realizada a fermentação em um biorreator de leito empacotado cilíndrico, sendo os gases provenientes do leito e conduzidos a um analisador de gases. Foi proposto um modelo matemático unidimensional, capaz de prever os perfis de temperatura e o crescimento fúngico ao longo do processo em qualquer posição do biorreator. Os resultados experimentais em embalagens plásticas e no biorreator mostraram que menores quantidades de substrato são favoráveis à maior estabilidade da temperatura do meio de cultivo, resultando em maiores quantidades de esporos. No teste de área superficial, foi possível observar um aumento na área da quirera de arroz e consequentemente aumento da quantidade de esporos. Nos testes de cinética de crescimento, pode se observar que o consumo de O2 é baixo comparado a outros fungos. Os dados de simulação não demonstraram sobreaquecimento do meio de cultivo no biorreator com o aumento de carga, o que não afetaria a produção de esporos. Também pode se observar com os dados de simulação que a temperatura da superfície do meio de cultivo não sofreu sobreaquecimento. Porém, com os dados experimentais, foi possível observar um sobreaquecimento no meio de cultivo, em virtude da geração de calor metabólico. Os dados experimentais demonstraram que os melhores resultados obtidos no biorreator, para produção de esporos do fungo Metarhizium anisopliae, foram com cargas de 1 Kg, correspondente a 2 cm de altura de substrato e que com cargas maiores os perfis de temperatura apresentaram elevado aquecimento no meio de cultivo. / This study aimed to develop a bioreactor tray capable of producing spores of the fungus Metarhizium anisopliae entomopathogenic. To this end, we used a tray bioreactor for the production of spores, which in turn was evaluated according to the employed substrate load. Temperature variations in the bioreactor were simulated and observed experimentally. For the development of bioreactor assays were carried out in plastic bags containing substrate 10g and 500g in order to evaluate how increasing fermentative medium loading could influence the production of spores. In these tests we measured the temperature at the geometric center of the packages and the temperature rise was related to spore production performed -. A surface area analysis of rice Type 1 and broken rice to evaluate the interference at this property in the production of spores. From the results in plastic containers experiments were performed in a bioreactor of 40 cm long, 29 cm wide by 12 cm, air flowing onto the particles in the direction of the largest dimension. Loads were applied to 1, 2 and 3 kg of rice type 1, corresponding to the bed height of 2, 4 and 6 cm, respectively, and 1 kg of grits, corresponding to 2 cm. The temperature at the inlet position and the air outlet and the geometric mean of the culture medium were measured throughout the tests. Fermentation in plastic packaging and the bioreactor were carried out at 28 C. To estimate the generation of metabolic heat of culture medium were collected consumption data The O2 and CO2 release during the incubation period. To this end, the fermentation was performed in a bioreactor cylindrical packed bed, and the gases from the bed and driven to a gas analyzer. A one-dimensional mathematical model capable of predicting the temperature profile and fungal growth during the process at any position of the bioreactor is proposed. The experimental results in plastic bottles and bioreactor showed that smaller amounts of substrate are conducive to increased stability of the temperature of the medium, resulting in larger amounts of spores. In the test surface area, it was possible to observe an increase in the area of rice grits and therefore increasing the amount of spores. In the growth kinetics tests, it can be seen that the O 2 consumption is low compared to other fungi. Simulation data showed no overheating of the culture medium in the bioreactor with the increased load, which would not affect the production of spores. It can also be observed with the simulation data as the temperature of the surface of the medium has not suffered overheating. However, with the experimental data, it was possible to observe overheating in the culture medium by virtue of the generation of metabolic heat. The experimental data show that the best results obtained in the bioreactor for the production of Metarhizium anisopliae spores of the fungus were to loads of 1 kg, corresponding to 2 cm substrate height and with larger loads temperature profiles showed high heat in the middle cultivation.

No período de 1999 a 2002, uma linhagem de Salmonella Enteritidis esteve envolvida em mais de 90% das salmoneloses ocorridas no RS. Este trabalho teve por objetivo avaliar a cinética de crescimento, a resistência ácida e a resistência térmica dessa linhagem e compará-la com S. Typhimurium e S. Bredeney não envolvidas em surtos alimentares, porém isoladas na mesma região. Em uma primeira etapa, a cinética de crescimento foi avaliada semeando-se cada sorovar em caldo nutriente (CN) e em salada de batata com maionese caseira (SMC), os quais foram mantidos a 30°C e 9,5°C. Em CN, a cinética de crescimento a 30°C foi semelhante para todos os sorovares, porém, em SMC a S. Enteritidis apresentou maior quantidade de células nas primeiras 6 horas de crescimento, sendo que somente depois de 12 horas todos os sorovares atingiram quantidades semelhantes de células. Em CN e em SMC, na temperatura de 9,5°C, não foi detectado crescimento de nenhum dos sorovares de Salmonella durante as primeiras 24 horas, sugerindo que essa temperatura foi suficiente para controlar a multiplicação desses microrganismos. Em uma segunda etapa, avaliou-se a resistência ácida e térmica dos diferentes sorovares de Salmonella. Para tanto, os três sorovares foram inoculados separadamente em CN e CN enriquecido com 1% de glicose (CNG), este último utilizado para produção de culturas ácido-adaptadas. Em seguida, os microrganismos foram submetidos a diferentes pH (3,5; 4,0 e 4,5) e temperaturas (52, 56 e 60ºC). Os resultados demonstraram que a S. Bredeney apresentou maior resistência para os pH 3,5 e 4,0, porém a S. Enteritidis demonstrou maior capacidade de adaptação ácida do que S. Typhimurium e S. Bredeney. Em pH 4,5 todos os sorovares, tanto não adaptados quanto ácido-adaptados, mantiveram a mesma quantidade de células viáveis durante 300 minutos. Quando expostas a 52ºC, S. Bredeney apresentou maior resistência, entretanto somente a S. Enteritidis foi protegida com a adaptação ácida. Para 56 e 60ºC, a S. Enteritidis, não adaptada e ácido-adaptada, apresentou maior resistência. A análise por SDS-PAGE demonstrou diferenças no perfil protéico de células não adaptadas e ácidoadaptadas para todos os sorovares testados. Com base nestes resultados, a capacidade de multiplicação mais rápida nas primeiras horas de cultivo em SMC, a maior capacidade de adaptação ácida e a maior resistência térmica demonstradas pela S. Enteritidis podem estar relacionadas ao freqüente envolvimento desse sorovar nas salmoneloses do RS. / During the period of 1999 to 2002, a strain of Salmonella Enteritidis was involved in more than 90 % of foodborne salmonellosis occurred in Rio Grande do Sul (RS) State. This work aimed to evaluate the growth kinetics, and the acid and the thermal resistance of this strain, comparing with S. Typhimurium and S. Bredeney, which were not involved in foodborne outbreaks, but were isolated in the same region. In the first stage of this study, the growth kinetics was assessed. Each serovar was inoculated separately in nutrient broth (CN) and in potato salad prepared with homemade mayonnaise (SMC), and then incubated at 30 ºC and 9.5 ºC. In CN, at 30 ºC, similar growing characteristics were found for all serovars, however in SMC S. Enteritidis demonstrated higher counts at the first 6 hours. Only after 12 hours of incubation, all serovars reached similar counts. In CN and in SMC, at 9.5 ºC, during the first 24 hours, there was no detectable growth of any Salmonella serovar, suggesting that such temperature was adequate to control the multiplication of tested Salmonella serovars. In the second stage of the study, the acid and the thermal resistances of Salmonella serovars were evaluated. The three serovars were cultivated separately in Nutrient Broth and Nutrient Broth supplemented with 1 % glucose (NBG). The latter medium was used to induce acid-adapted cells. Then, the three serovars were exposed to different pH (3.5, 4.0, and 4.5) and temperatures (52, 56, and 60 ºC). Results indicated that S. Bredeney presented higher resistance to pH 3.5 and 4.0, but S. Enteritidis presented a better capacity of acid adaptation than S. Typhimurium and S. Bredeney. At pH 4.5, all serovars demonstrated a similar behavior, remaining at same levels of viable cells until 300 minutes. At 52 ºC, S. Bredeney presented greater survivor rates, however acid adaptation protected only S. Enteritidis. At 56 ºC and 60 ºC, non-adapted and acidadapted S. Enteritidis were more thermally resistant than other serovars tested. SDS-PAGE analysis demonstrated differences in protein profile of non-adapted and acid-adapted cells of all serovars. The capacity of rapid multiplication in the first hours of cultivation, the greater acid adaptation and thermal resistance presented by S. Enteritidis, may be related to the frequent involvement of this strain in salmonellosis cases in the RS.

abstract: Photosynthesis converts sunlight to biomass at a global scale. Among the photosynthetic organisms, cyanobacteria provide an excellent model to study how photosynthesis can become a practical platform of large-scale biotechnology. One novel approach involves metabolically engineering the cyanobacterium Synechocystis sp. PCC 6803 to excrete laurate, which is harvested directly. This work begins by defining a working window of light intensity (LI). Wild-type and laurate-excreting Synechocystis required an LI of at least 5 µE/m2-s to sustain themselves, but are photo-inhibited by LI of 346 to 598 µE/m2-s. Fixing electrons into valuable organic products, e.g., biomass and excreted laurate, is critical to success. Wild-type Synechocystis channeled 75% to 84% of its fixed electrons to biomass; laurate-excreting Synechocystis fixed 64 to 69% as biomass and 6.6% to 10% as laurate. This means that 16 to 30% of the electrons were diverted to non-valuable soluble products, and the trend was accentuated with higher LI. How the Ci concentration depended on the pH and the nitrogen source was quantified by the proton condition and experimentally validated. Nitrate increased, ammonium decreased, but ammonium nitrate stabilized alkalinity and Ci. This finding provides a mechanistically sound tool to manage Ci and pH independently. Independent evaluation pH and Ci on the growth kinetics of Synechocystis showed that pH 8.5 supported the fastest maximum specific growth rate (µmax): 2.4/day and 1.7/day, respectively, for the wild type and modified strains with LI of 202 µE/m2-s. Half-maximum-rate concentrations (KCi) were less than 0.1 mM, meaning that Synechocystis should attain its µmax with a modest Ci concentration (≥1.0 mM). Biomass grown with day-night cycles had a night endogenous decay rate of 0.05-1.0/day, with decay being faster with higher LI and the beginning of dark periods. Supplying light at a fraction of daylight reduced dark decay rate and improved overall biomass productivity. This dissertation systematically evaluates and synthesizes fundamental growth factors of cyanobacteria: light, inorganic carbon (Ci), and pH. LI remains the most critical growth condition to promote biomass productivity and desired forms of biomass, while Ci and pH now can be managed to support optimal productivity. / Dissertation/Thesis / Doctoral Dissertation Civil and Environmental Engineering 2015

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-03-09T20:25:06Z No. of bitstreams: 1 BRUNA DA SILVEIRA GUIMARÃES - DISSERTAÇÃO PPGEQ 2016..pdf: 3845611 bytes, checksum: 4e146e7310b6d3800c9b8ead6562e7a6 (MD5) / Made available in DSpace on 2018-03-09T20:25:06Z (GMT). No. of bitstreams: 1 BRUNA DA SILVEIRA GUIMARÃES - DISSERTAÇÃO PPGEQ 2016..pdf: 3845611 bytes, checksum: 4e146e7310b6d3800c9b8ead6562e7a6 (MD5) Previous issue date: 2016-04-15 / Capes / As microalgas são microrganismos unicelulares que podem crescer tanto em água doce como salgada. São clorofilados e estão em praticamente todos os tipos de água do planeta. Seu cultivo vem cada vez mais despertando o interesse, já que a aplicabilidade vem sendo observada em vários segmentos tais como no tratamento de água, nutrição e na geração de energia. A microalga Scenedesmus sp., é clorofilada, unicelular e uninucleada e pertencem à família Scenedesmaceae, gênero Scenedesmus. Tal microalga é mais comum em água doce, podendo ser encontrada com menor freqüência em águas salobras, com salinidade entre 0,05 e 3% de sais (500 a 30000 ppm). Possuem forma elipsoidal, colônias lineares e agrupadas em números de 4, 8, 16 ou 32. Este trabalho teve como objetivo o estudo da cinética de crescimento da microalga Scenedesmus sp. em meios salobros, através produção de dois meios de cultivo sintéticos com adição dos sais NaCl e MgCl2, visando a possibilidade de posteriores aproveitamentos de águas salobras para a produção de microalgas. Foi verificado se a presença dos sais tem influência no crescimento de tais microrganismos. Analisando as curvas de crescimento microbiano da microalga Scenedesmus sp., observou-se que nos meios contendo o cloreto de magnésio a fase Lag ou de latência praticamente não ocorreu, com exceção do meio com concentração de 8000 mg.L-1, indicando que a microalga demorou menos tempo para se adaptar ao meio com adição de MgCl2, se comparado com os meios contendo NaCl. Constatou-se também que a influência do íon cloreto no crescimento da microalga foi diminuída devido à presença dos íons sódio e magnésio, que são nutrientes importantes no metabolismo celular. Analisando os parâmetros cinéticos calculados (velocidade máxima de crescimento, produtividade em biomassa e tempo de geração), observou-se que os melhores resultados foram obtidos nos meios contendo cloreto de magnésio, indicando uma melhor cinética de crescimento nestes meios, se comparado com os que continham NaCl. Isto ocorreu devido ao fato de que o Mg é classificado como um macronutriente ou nutriente essencial, importante para o metabolismo celular da microalga e o sódio um micronutriente (necessário em menores quantidades), sendo mais observada a influência daquele íon no crescimento da microalga do que dos íons de sódio. Baseado nos resultados cinéticos obtidos, constatou-se a possibilidade de crescimento da microalga Scenedesmus sp. em meios salobros. / Microalgae are unicellular microorganisms that can grow in both freshwater and saltwater. They are chlorophyll and are practically in all kinds of water on the planet. Its cultivation is increasingly attracting interest since the applicability has been observed in several segments, such as in the treatment of water, nutrition and energy generation. The microalgae Scenedesmus sp. is chlorophyllated, unicellular and uninucleate and belongs to Scenedesmaceae family, genus Scenedesmus. This microalgae is more common in fresh water and can be found less frequently in brackish water with salinity between 0.05 and 3% salts (500-30000 ppm). It has ellipsoidal shape, linear colonies and grouped in numbers 4, 8, 16 or 32. This study aimed growth kinetics study of microalgae Scenedesmus sp. in brackish means, by producing two synthetic culture media with the addition of NaCl and MgCl2 salts, with a view to possible subsequent utilizations of brackish water for the production of microalgae. It has been found that the presence of salts influences the growth of such microorganisms. Analyzing the microbial growth curves of the microalgae Scenedesmus sp., it was noted that, in the means containing magnesium chloride, the Lag phase or latency hardly occurred, with the exception of media with concentration of 8000 mg.L-1, indicating that the microalgae took less time to adapt to the medium with addition of MgCl2, compared with media containing NaCl. It was also found that the influence of chloride ion in the growth of microalgae was decreased due to the presence of sodium and magnesium ions, nutrients that are important in cellular metabolism. By analyzing the calculated kinetic parameters (maximum growth rate, productivity of biomass and generation time), it was observed that the best results were obtained in media containing magnesium chloride, indicating a better growth kinetics in these media compared to those containing NaCl. This was due to the fact that Mg is classified as a macronutrient or essential nutrient, important for cell metabolism of microalgae, and sodium is classified as a micronutrient (required in smaller amounts). The influence of that ion (Mg) in microalgae growing has been more observed than sodium ions influence. Based on the kinetic results, the possibility of the microalga Scenedesmus sp growth. in brackish means has been verified.

Mesenchymal stem cells (MSCs) derived from human adipose tissue and umbilical cord (Wharton’s jelly, UCB) represent a useful source of adult stem cells for cellular therapy and tissue engineering. The biggest concern with the use of MSCs therapeutically relates to their isolation and growth/manipulation ex vivo. This study aimed to establish methods for the routine isolation and characterization of MSCs from human tissues. The objectives were (1) to show that MSCs could be isolated from different human tissues, namely adipose tissue, Wharton’s jelly, and UCB; (2) to confirm the MSC phenotypic profile over at least 10 passages; and (3) to show the multilineage differentiation capacity of the isolated cells. The minimal criteria as defined by the International Society for Cellular Therapy (ISCT) were used to determine whether MSCs were successfully isolated from various human tissues. Two different techniques involving enzymatic digestion or explant cultures were utilized, and compared for isolating MSCs from Wharton’s jelly. Umbilical cord blood has been suggested as another source of MSCs. However, we were unable to grow MSCs from UCB. Proliferation kinetics of isolated MSCs revealed that cords, either from digested cords or cord pieces had a mean PDT from passage 1 to 4 that was approximately 3 fold lower than for the ASCs. Mesenchymal stem cells from adipose tissue and Wharton’s jelly expressed the classical MSC phenotype (CD73+, CD90+, CD105+, CD34-, and CD45-). The cells from Wharton’s jelly showed a more uniform MSC profile over passages, with higher levels of marker expression when compared to ASCs. Variability in phenotype was observed in early ASC passages, whereas WJ-MSCs seemed to attain the MSC phenotype as early as passage 0 for both isolation techniques. Low levels of CD34 positive cells remained in the ASCs. Oil red O staining was used for identifying the lipid droplets in adipogenic differentiation cultures. A colorimetric assay as well as image analysis was used to quantify the differentiation. For the cord samples, both assays produced positive results. Histological examination, however, revealed that the cords did not form lipid droplets. The ASCs showed a statistically significantly greater differentiation capacity into adipocytes compared with the cords (pooled digested and pieces data). Alizarin red S staining was used for identifying calcium deposition during matrix mineralization in osteogenic differentiation cultures. No significant differences in osteogenic differentiation were observed between ASCs and WJ-MSCs. Chondrogenic differentiation was observed for both MSC sources by positive staining of glycosaminoglycans using toluidine blue O. The main findings of the study showed that MSCs, according to the ISCT guidelines, were successfully harvested from adipose tissue. However, due to the lack of adipogenic differentiation of WJ-derived cells, they did not meet the ISCT guidelines to be classified as MSCs, and were referred to as MSC-like cells. Regardless of the isolation technique used, Wharton’s jelly yielded cells with similar proliferation capacity, phenotype, and differentiation capacity. This study did, however, reveal that biological differences do exist between stem cells from different sources. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Immunology / unrestricted

The crystal growth of nanoporous materials is different from most other classes of material in that their framework architectures contain periodic arrangement of pores or voids in which there is no direct bonding between adjacent units of the framework. This poses a variety of questions as to how such parts of framework develop during the crystallization process, atomistically and energetically. Here we use the nanoporous metal-organic framework, ZIF-8 as a prototypical material to obtain a basic understanding of the growth of a nanoporous material. The crystals of ZIF-8 produced in the N,N-dimethylformamide solvent [ZIF-8(DMF)] and methanol-co-N,N-dimethylformamide solvent [ZIF-8(MeOH)] are both rhombic dodecahedron in shape with a much smaller crystal size in the latter. In the study of the kinetics of ZIF-8(DMF) crystallization, we get a good agreement in the values of activation energies using both Avrami-Erofe’ev-Hancock-Sharp and Gualtieri’s models, i.e. about 120 kJ mol-1 for nucleation, and 95 kJ mol-1 for crystal growth process. The study of kinetics of ZIF-8 surface growth, by in situ AFM, with ZIF-8(DMF) as seed crystal that are grown in the methanolic growth solution we see faster rate in the <100> directions than the <110> directions, with the most probable activation energy of about 80 kJ mol-1 in both directions. This is the first example of in situ AFM being used to obtain activation energy for a surface growth in MOF. We also reveal here that growth process of ZIF-8 occurs through the nucleation and spreading of successive metastable unenclosed sub-steps to eventually form stable terrace steps of the enclosed framework structure in which this process is reliant on the presence of nonframework species to connect the framework species that have voids between them. The experiments also enable identification of some of the fundamental units in the growth process and the stable crystal surface plane. Further, the spreading of terraces at high supersaturation condition (early state) is fairly isotropic as is seen through the formation of almost-rounded terraces on the surface of ZIF-8. The growth direction becomes clear as the supersaturation condition nears to equilibrium (later stage) by the formation of rhombohedral terraces with pointy ends growing along the <100>, and <110> directions and straight edges growing perpendicular to the <111> direction. Formation of this rhombohedral morphology is explained by a coarse grain approach similar to that used in the Kossel model by making assumptions that the sodalite cage is the growth unit and attachment of one sodalite cage in each growth direction is the rate determining step for the formation of a new row of sodalite cages in each direction. Finally, based on the profiles of growth spirals formed from screw dislocations on the ZIF-8 surface obtained from the ex situ AFM images and ICE theory, plausible screw dislocations with Burgers’ vector of 1/2 <111> and <100>, but not <110>, are deduced. Some of the findings in this work will be applicable to numerous nanoporous materials, and the work in general will support efforts to synthesize and design new framework materials and to control the crystal properties of these materials.