La cristallisation de la cire dans les huiles pétrolières est un processus qui se produit à la suite de changements dans les conditions de pression et de température dans lesquelles ils sont sur le gisement, et qui diminuent progressivement le long de ses processus de production, transport et de stockage. Sa précipitation et son dépôt sur les surfaces intérieures des pipelines nécessitent la mise en œuvre des techniques de guérison ou d'amélioration des propriétés des fluides, afin de réduire les pertes causées par leur accumulation, qui dans certains cas pourrait complètement paralyser le production d'une plante. L'étude du processus de précipitation des paraffines est basée sur la détermination des conditions de pression et de température dans lesquelles il se produit, et la caractérisation des phases coexistent en équilibre. Parmi les principales limites de ce processus, nous trouvons de la disponibilité et de la conception des appareils expérimentaux qui permettent la détection des températures de fusion finissantes et la séparation de phases sous pression, et le développement des méthodes chromatographiques pour l'identification et la quantification des paraffines lourdes. Le développement de modèles thermodynamiques qui décrivent le processus de précipitation des paraffines sont l'un des principaux outils pour l'anticipation et la prévention des processus de cristallisation des cires. Son efficacité dans l'estimation des conditions dans lesquelles les cristaux commencent à apparaître, ainsi que la quantité totale de solides qui pourrait être formée, dépendent de la conception des techniques expérimentales qui reproduisent les conditions réelles d'opération, et de recueillir des informations fiables du comportement de phase dans des mélanges des hydrocarbures. Dans ce travail de recherche s'est développé une méthodologie complète pour la détection et filtration des cristaux dans une gamme s'étendant à partir des conditions normales jusqu'à 800 bars et une méthode chromatographique d’haute température qui ont permis d'effectuer une évaluation du comportement de phase en fonction de la composition, la pression et la température, ainsi que l'identification et la quantification des paraffines. Le comportement de phase des mélanges de paraffines étudiés indiquent que les variations de pression et de température ont une grande influence sur la cristallisation des cires: les processus de compression isotherme refroidissement isobare diminuent la solubilité des cires et entraîner non seulement dans une augmentation de la quantité de solides précipités, mais dans une variante de sa nature. De plus, l'augmentation de la pression du système provoque une augmentation linéaire des températures de changement de phase des mélanges étudiés. Pour sa part, les différences de composition des mélanges étudiés ont généré des différences significatives dans leur température de changement de phase, mais pas dans la quantité de matières solides cristallisées. En général, des mélanges enrichis en paraffines lourdes ont produit une augmentation de leur WAT. Les résultats obtenus ont montré que l'effet prédominant sur le processus de cristallisation des cires correspond à la diminution de la température du système, ce qui conduit à la formation de quantités importantes de déchets solides. De plus, elles sont consistant avec le comportement de phase des mélanges de cires synthétiques, ce qui permet de conclure que la méthode expérimentale développé, constitué par le couplage de la technique de détection et de séparation de phase avec chromatographie en phase gazeuse à haute température, représente outil précis et fiable pour l'étude des processus de précipitation de la paraffine dans des conditions réelles d'opération, et qui a surmonté les limites expérimentales typiques de ces analyses. / The wax crystallization in oil fluids is a process that occurs as a result of changes in pressure and temperature conditions found in the site, and gradually decreasing along the production process, transport and storage thereof. Precipitation and deposition on the inner surfaces of pipelines require the implementation of healing techniques or to improve the properties of fluids, in order to reduce losses caused by their accumulation, which in some cases could come to a complete standstill production plant. The study of paraffin precipitation process is based on determining the pressure and temperature conditions in which it occurs, and the characterization of the phases coexist in equilibrium. The main constraints of this process, we found the availability and experimental design devices that allow detection of the onset temperatures of crystals and phase separation at high pressure and the development of chromatographic methods for the identification and quantification of paraffins high molecular weight. The development of thermodynamic models describing the precipitation of paraffins is one of the main tools for anticipating and preventing wax crystallization process. Its effectiveness in estimating the conditions that will start appearing crystals and the total amount of solid that could be formed, depends on the design of experimental techniques to reproduce the real operating conditions, and gather reliable information of phase behavior in mixtures hydrocarbon. In the present investigation a complete methodology for detecting and filtering crystals in a range extending to 800 bar normal conditions and a high temperature chromatographic method that allowed to carry out the assessment phase behavior developed in Depending on the composition, pressure and temperature as well as the identification and quantification of heavy paraffins. Phase behavior of mixtures of paraffins studied indicates that changes in pressure and temperature have a great influence on the crystallization of waxes: isothermal compression processes isobaric cooling and decrease the solubility of the wax and result not only in an increase in the amount of precipitated solids but in a variation of the nature thereof. Additionally, increasing the system pressure generated an increase in temperature of the phase change mixtures studied. For its part, the differences in composition of the mixtures studied generated significant differences in their phase change temperature but not in the amount of solid crystallized. In general, enriching the mixture to heavy paraffins produced an increase in their respective WAT. The results obtained showed that the predominant effect in the wax crystallization process corresponds to the temperature decrease of systems, leading to the formation of large quantities of solid waste. Also, these are consistent with the phase behavior of mixtures of synthetic waxes, which allows to conclude that the experimental methodology developed, consisting of the coupling and detection techniques phase separation gas chromatography with high temperature, represents accurate and reliable tool for the study of paraffin precipitation process in real operating conditions, and who overcame the typical experimental limitations of these analyzes.

<p>The interest for miniaturisation in chemical and biological analysis has increased in recent years. In this work, the design, fabrication and characterisation of tools for microanalysis have been studied. The focus is set on polymer microchips for applications in chemical analysis. The work consists of three parts: design and fabrication of paraffin microactuators, design and fabrication of polymer microchips as interfaces in electrospray ionisation mass spectrometry (ESI-MS), and characterisation of conducting films for fused silica capillaries as interfaces in ESI-MS.</p><p>The principle of the paraffin actuators is based on the volume increase resulting from paraffin melting. Paraffin expansion is utilised to cause membrane deflection. The first plastic microactuator using paraffin as the actuator material was successfully demonstrated.</p><p>The microchips as interfaces in ESI-MS have been designed with the objective that the interface should be as much a part of the microchip as possible, and as to as large extent as possible, be fabricated in the same step as the microchannels. Sheathless electrospray from microchips was demonstrated for the first time. In addition a simplified fabrication process for ESI-MS interfaces in poly(dimethyl siloxane) (PDMS) was developed.</p><p>The degradation of conductive coatings for sheathless ESI-MS on fused silica capillaries was studied. It was shown that electrochemical experiments could successfully be used to simulate the electrospray conditions and predict the failure of different gold coatings.</p><p>It was concluded that a common issue in the fabrication of thermoplastic microchips is the crucial sealing of microchannels and cavities. From this point of view, PDMS is a more advantageous material in microfluidics.</p>

When paraffin wax melts it exhibits a large, relatively incompressible volume expansion. This can be used in microactuators for strong and large displacements, a rare combination among actuators. Furthermore, paraffin is inexpensive, inert and environmentally friendly, as well as easily processed and actuated. Together, these properties give paraffin actuators great potential for use in both low-cost and high-performance applications. In microfluidics, the miniaturization of various analysis systems decreases the volumes of samples and reagents needed, as well as the analysis throughput time. Using on-chip micropumps increases the efficiency of the microfluidic system, but a challenge for such pumps is the high back-pressure associated with separation, filtration or narrower channels. The objective of this thesis is to increase the understanding of paraffin in microactuators, as well as to further explore its possibilities and limitations. The main application area has been on-chip micropumps. For low-cost applications, actuators, pumps and dispensers have been fabricated in plastics and then evaluated. The dispenser is intended for on-chip storage and dispensing of liquids in a lab-on-a-chip that could be used in, e.g., point-of-care testing (POCT). For high-performance applications, metallic actuators, pumps and dispensers have been accomplished. The micropump is the world’s strongest mechanical micropump in sub-cubic centimetre size, capable of pressures of above 5 MPa. Possible applications are strong microhydraulics, on-chip chromatography, or medical microdosage systems. A limitation of paraffin is the relatively slow thermal actuation. In this thesis the thermal properties have also been turned into an advantage: Directional solidification is used to accomplish multiple stable states of the actuator displacement, withheld without any power consumption. For the future, the high-pressure capability may be improved by new designs. Optimization of speed and power consumption can be made by further work on modelling as well as on drive and control of the heating.

The interest for miniaturisation in chemical and biological analysis has increased in recent years. In this work, the design, fabrication and characterisation of tools for microanalysis have been studied. The focus is set on polymer microchips for applications in chemical analysis. The work consists of three parts: design and fabrication of paraffin microactuators, design and fabrication of polymer microchips as interfaces in electrospray ionisation mass spectrometry (ESI-MS), and characterisation of conducting films for fused silica capillaries as interfaces in ESI-MS. The principle of the paraffin actuators is based on the volume increase resulting from paraffin melting. Paraffin expansion is utilised to cause membrane deflection. The first plastic microactuator using paraffin as the actuator material was successfully demonstrated. The microchips as interfaces in ESI-MS have been designed with the objective that the interface should be as much a part of the microchip as possible, and as to as large extent as possible, be fabricated in the same step as the microchannels. Sheathless electrospray from microchips was demonstrated for the first time. In addition a simplified fabrication process for ESI-MS interfaces in poly(dimethyl siloxane) (PDMS) was developed. The degradation of conductive coatings for sheathless ESI-MS on fused silica capillaries was studied. It was shown that electrochemical experiments could successfully be used to simulate the electrospray conditions and predict the failure of different gold coatings. It was concluded that a common issue in the fabrication of thermoplastic microchips is the crucial sealing of microchannels and cavities. From this point of view, PDMS is a more advantageous material in microfluidics.

Um dos grandes desafios enfrentados pela indústria do petróleo é reduzir o impacto causado pela cristalização indesejável de hidrocarbonetos parafínicos de elevada massa molar em tubulações e equipamentos de produção. A cristalização de parafinas em petróleo é normalmente detectada através da determinação da temperatura inicial de aparecimento de cristais (TIAC), que pode ser estimada através de modelagem termodinâmica com base na composição do petróleo. Os objetivos deste trabalho são: estudar os principais modelos termodinâmicos adotados para descrever a precipitação de parafinas; verificar a validade desses modelos para os petróleos brasileiros e determinar qual modelo é o mais adequado para esses óleos. Para tanto, três formas de cálculo da razão entre as fugacidades das fases sólida e líquida e cinco modelos para calcular os coeficientes de atividade dos componentes em cada fase são aplicados aos dados de composição de vinte e três petróleos brasileiros. Os resultados mostram que o modelo ideal de múltiplas fases sólidas e o modelo de Escobar-Remolina geram valores bastante abaixo da TIAC experimental. Para os modelos de única fase sólida ideal, de Won e de Coutinho, foi possível observar que: a) a grande maioria dos erros é negativa; b) que estes se distribuem melhor em torno de zero quando se utiliza a correlação de Coutinho para o cálculo dos valores de &#61529;i; c) os valores de erro médio para os modelos de Coutinho, de Won e ideal com única fase sólida se equivalem, qualquer que seja o modelo utilizado para o cálculo de &#61529;i, exceto para o caso em que todos os compostos presentes na fase líquida podem precipitar; d) os valores obtidos através do modelo de Coutinho apresentam erro sistemático em relação ao modelo ideal de única fase sólida; e) as diferentes formas de se calcular a razão entre as fugacidades da fase sólida e líquida (&#61529;i) influenciam fortemente a capacidade preditiva dos modelos, o que não era esperado; f) o perfil do primeiro cristal formado nos petróleos é influenciado pelas moléculas mais pesadas presentes nos resíduos, o que mostra a necessidade de se desenvolver metodologias precisas e robustas de caracterização de resíduos; g) a inclusão de uma estimativa para a composição dos resíduos efetivamente melhorou o desempenho dos modelos em petróleos médio; h) em petróleos pesados, houve um aumento do erro de previsão da TIAC devido à pouca ou nenhuma quantidade de parafinas nos resíduos desses óleos. A necessidade de uma melhor caracterização dos resíduos de petróleos é corroborada pelo fato da TIAC calculada pelos modelos ser, via de regra, mais baixa que a TIAC experimental e pela melhora no desempenho dos modelos quando se estimou a composição dos resíduos, em petróleos médios / A major challenge faced by the oil industry is to reduce the impact of undesirable crystallization of high molecular weight paraffins in pipelines and production equipments. The paraffin crystallization is usually detected by measuring the wax appearance temperature (WAT), which can be estimated from composition based thermodynamic modeling. The goals of this work are: to study the main thermodynamic models adopted for paraffin crystallization description, to check models performances for Brazilian crudes and to determine the best model for these oils. For this, composition data from twenty-three Brazilian oils were used. For the thermodynamic modeling of wax precipitation in these crude oils, three forms of calculating the ratio between the solid and the liquid phase fugacities and five models to calculate the activity coefficients of components in each phase were used. The results showed that the ideal model for multiple solid phases and the Escobar-Remolina model give results far below the experimental WAT. For the single solid ideal, Won and Coutinho models, the points to be mentioned are described as follows: a) most of the errors is negative; b) the errors distribution is better when the correlation used for &#61529;i calculation is the number 2; c) the average errors for ideal, Won and Coutinho models are equivalent, except when all the oil components are considered to precipitate; d) the results obtained from Coutinho model presents systematic error in relation to the ideal model; e) the different ways to calculate the ratio between the solid and the liquid phase fugacities strongly influence the predictive ability of models, which was not expected; f) the composition profile of the first crystal formed in the oil is influenced by the heavier molecules in the residue, which shows the need of developing robust and accurate methods of heavy fractions characterization; g) the inclusion of an estimate for the residue composition effectively improved the performance of models for intermediate crude; h) for heavy oil, there was an increase in prediction error due to the little amount of paraffinic compounds in these residues. The need for better characterization of the residues is corroborated by the fact that the calculated WAT is normally lower than the experimental WAT and by the improvement on the models performance when the estimation of the residue composition was done for intermediate crudes

Um dos grandes desafios enfrentados pela indústria do petróleo é reduzir o impacto causado pela cristalização indesejável de hidrocarbonetos parafínicos de elevada massa molar em tubulações e equipamentos de produção. A cristalização de parafinas em petróleo é normalmente detectada através da determinação da temperatura inicial de aparecimento de cristais (TIAC), que pode ser estimada através de modelagem termodinâmica com base na composição do petróleo. Os objetivos deste trabalho são: estudar os principais modelos termodinâmicos adotados para descrever a precipitação de parafinas; verificar a validade desses modelos para os petróleos brasileiros e determinar qual modelo é o mais adequado para esses óleos. Para tanto, três formas de cálculo da razão entre as fugacidades das fases sólida e líquida e cinco modelos para calcular os coeficientes de atividade dos componentes em cada fase são aplicados aos dados de composição de vinte e três petróleos brasileiros. Os resultados mostram que o modelo ideal de múltiplas fases sólidas e o modelo de Escobar-Remolina geram valores bastante abaixo da TIAC experimental. Para os modelos de única fase sólida ideal, de Won e de Coutinho, foi possível observar que: a) a grande maioria dos erros é negativa; b) que estes se distribuem melhor em torno de zero quando se utiliza a correlação de Coutinho para o cálculo dos valores de &#61529;i; c) os valores de erro médio para os modelos de Coutinho, de Won e ideal com única fase sólida se equivalem, qualquer que seja o modelo utilizado para o cálculo de &#61529;i, exceto para o caso em que todos os compostos presentes na fase líquida podem precipitar; d) os valores obtidos através do modelo de Coutinho apresentam erro sistemático em relação ao modelo ideal de única fase sólida; e) as diferentes formas de se calcular a razão entre as fugacidades da fase sólida e líquida (&#61529;i) influenciam fortemente a capacidade preditiva dos modelos, o que não era esperado; f) o perfil do primeiro cristal formado nos petróleos é influenciado pelas moléculas mais pesadas presentes nos resíduos, o que mostra a necessidade de se desenvolver metodologias precisas e robustas de caracterização de resíduos; g) a inclusão de uma estimativa para a composição dos resíduos efetivamente melhorou o desempenho dos modelos em petróleos médio; h) em petróleos pesados, houve um aumento do erro de previsão da TIAC devido à pouca ou nenhuma quantidade de parafinas nos resíduos desses óleos. A necessidade de uma melhor caracterização dos resíduos de petróleos é corroborada pelo fato da TIAC calculada pelos modelos ser, via de regra, mais baixa que a TIAC experimental e pela melhora no desempenho dos modelos quando se estimou a composição dos resíduos, em petróleos médios / A major challenge faced by the oil industry is to reduce the impact of undesirable crystallization of high molecular weight paraffins in pipelines and production equipments. The paraffin crystallization is usually detected by measuring the wax appearance temperature (WAT), which can be estimated from composition based thermodynamic modeling. The goals of this work are: to study the main thermodynamic models adopted for paraffin crystallization description, to check models performances for Brazilian crudes and to determine the best model for these oils. For this, composition data from twenty-three Brazilian oils were used. For the thermodynamic modeling of wax precipitation in these crude oils, three forms of calculating the ratio between the solid and the liquid phase fugacities and five models to calculate the activity coefficients of components in each phase were used. The results showed that the ideal model for multiple solid phases and the Escobar-Remolina model give results far below the experimental WAT. For the single solid ideal, Won and Coutinho models, the points to be mentioned are described as follows: a) most of the errors is negative; b) the errors distribution is better when the correlation used for &#61529;i calculation is the number 2; c) the average errors for ideal, Won and Coutinho models are equivalent, except when all the oil components are considered to precipitate; d) the results obtained from Coutinho model presents systematic error in relation to the ideal model; e) the different ways to calculate the ratio between the solid and the liquid phase fugacities strongly influence the predictive ability of models, which was not expected; f) the composition profile of the first crystal formed in the oil is influenced by the heavier molecules in the residue, which shows the need of developing robust and accurate methods of heavy fractions characterization; g) the inclusion of an estimate for the residue composition effectively improved the performance of models for intermediate crude; h) for heavy oil, there was an increase in prediction error due to the little amount of paraffinic compounds in these residues. The need for better characterization of the residues is corroborated by the fact that the calculated WAT is normally lower than the experimental WAT and by the improvement on the models performance when the estimation of the residue composition was done for intermediate crudes

Durante as atividades de produção de petróleo offshore são utilizadas tubulações que transportam o petróleo bruto do fundo do poço até à plataforma ou até à costa marítima. No fundo mar as tubulações são submetidas a temperaturas da ordem de 4 oC. Nessas condições, quando a atividade de produção é interrompida para manutenções ou por paradas de emergência, o óleo bruto, com alto teor de parafinas, tende a se gelificar. Para reiniciar o escoamento são necessárias pressões acima da pressão usual de trabalho para promover a quebra do gel. A literatura mostra que não só o histórico de térmico, mas também o histórico de cisalhamento afetam a gelificação de óleos parafínicos e, portanto, a pressão de reinício do escoamento. O objetivo deste trabalho é avaliar experimentalmente as variáveis que influenciam o reinício do escoamento de petróleo parafínico. Para isso, uma unidade experimental foi construída de modo a reproduzir experimentalmente o reinício do escoamento de petróleo parafínico gelificado. A unidade permite o controle da temperatura, da taxa de resfriamento, da taxa de cisalhamento e do tempo de repouso na formação do gel. A pressão máxima necessária para o início do escoamento foi observada, os valores coletado e analisado. A investigação realizada mostra que todos os parâmetros testados apresentam alguma influência na pressão durante o reinício do escoamento de óleo parafínicos gelificados. / In offshore production, pipelines are used to transport crude oil from the well bore to the platform and from the plataform to the seacoast. When production is interrupted for maintenance or emergency, waxy crude oils tend to form gel at seabed low temperatures. At flow start-up, an increase in pressure above the usual operating pressure is required to break the gel. The literature has reported that not only the temperature but also the cooling rate, the shear rate during cooling and the material aging time affect the gelation of waxy crude oils, and, therefore, the start-up pressure. The objective of the current work is to evaluate experimentally the start-up flow of waxy crude oils in pipelines. In order to accomplish that, an experimental lab loop was build to reproduce the flow start-up of gelled waxy crude oils. The loop allows the control of temperature, cooling rate, shear rate and aging time in the gel formation. The pressure peaks measured during the flow restart were presented and analized as a function of the controlled parameters. The research shows that all parameters tested have some influence on the picks of pressure during the start up flow of paraffinic oil gelled.

Made available in DSpace on 2014-12-17T15:01:17Z (GMT). No. of bitstreams: 1 ErikaaAS.pdf: 744693 bytes, checksum: 969bbaf028a15425563a34bdced20b1e (MD5) Previous issue date: 2005-07-21 / The formation of paraffin deposits is common in the petroleum industry during production, transport and treatment stages. It happens due to modifications in the thermodynamic variables that alter the solubility of alkanes fractions present in petroleum. The deposition of paraffin can provoke significant and growing petroleum losses, arriving to block the flow, hindering to the production. This process is associated with the phases equilibrium L-S and the stages and nucleation, growth and agglomeration the crystals. That process is function of petroleum intrinsic characteristics and temperature and pressure variations, during production. Several preventive and corrective methods are used to control the paraffin crystallization, such as: use of chemical inhibitors, hot solvents injection, use of termochemistry reactions, and mechanical removal. But for offshore exploration this expensive problem needs more investigation. Many studies have been carried through Wax Appearance Temperature (WAT) of paraffin; therefore the formed crystals are responsible for the modification of the reologics properties of the oil, causing a lot off operational problems. From the determination of the WAT of a system it is possible to affirm if oil presents or not trend to the formation of organic deposits, making possible to foresee and to prevent problems of wax crystallization. The solvent n-paraffin has been widely used as fluid of perforation, raising the production costs when it is used in the removal paraffin deposits, needing an operational substitute. This study aims to determine the WAT of paraffin and the interference off additives in its reduction, being developed system paraffin/solvent/surfactant that propitiates the wax solubilization. Crystallization temperatures in varied paraffin concentrations and different solvents were established in the first stage of the experiments. In the second stage, using the methodology of variation of the photoelectric signal had been determined the temperature of crystallization of the systems and evaluated the interferences of additives to reduction of the WAT. The experimental results are expressed in function of the variations of the photoelectric signals during controlled cooling, innovating and validating this new methodology to determine WAT, relatively simple with relation the other applied that involve specific equipments and of high cost. Through the curves you differentiate of the results had been also identified to the critical stages of growth and agglomeration of the crystals that represent to the saturation of the system, indicating difficulties of flow due to the increase of the density / A forma??o de dep?sito paraf?nico ? comum na ind?stria do petr?leo durante as etapas de produ??o, movimenta??o e tratamento e ocorre em decorr?ncia de modifica??es nas vari?veis termodin?micas que alteram a solubilidade das fra??es de alcanos presentes no petr?leo. A cristaliza??o de parafinas pode provocar significativas e crescentes perdas de petr?leo, chegando a bloquear o fluxo, impedindo ? produ??o. Esse processo est? associado ao equil?brio de fases L-S e as etapas de nuclea??o, crescimento e aglomera??o dos cristais. Existem v?rios m?todos preventivos e corretivos para controlar a cristaliza??o de parafinas, dentre os quais se destacam: o uso de inibidores qu?micos, a inje??o de solventes aquecidos, o emprego de rea??es termoqu?micas e a remo??o mec?nica, por?m quanto ? explora??o offshore pouco se conhece, tornando-se indispens?vel investigar este custoso problema. Muitos estudos t?m sido realizados quanto ? temperatura de in?cio do aparecimento dos cristais (TIAC) da parafina, pois os cristais formados s?o respons?veis pela modifica??o das propriedades reol?gicas do petr?leo, causando diversos problemas operacionais. A partir da determina??o da TIAC de um sistema ? poss?vel afirmar se um petr?leo apresenta ou n?o tend?ncia ? forma??o de dep?sitos org?nicos, possibilitando prever e evitar problemas de cristaliza??o paraf?nica. O solvente n-parafina tem sido amplamente utilizado como fluido de perfura??o, elevando os custos de produ??o quando ? empregado na remo??o de dep?sitos paraf?nicos, necessitando de um substituto operacional. Este estudo visa determinar a TIAC da parafina e ? interfer?ncia de aditivos na sua redu??o, desenvolvendo um sistema parafina/solvente/tensoativo que propicie a solubiliza??o paraf?nica. Na primeira etapa dos experimentos, foram determinadas as temperaturas de cristaliza??o em concentra??es de parafina variadas e diferentes solventes. Na segunda etapa, utilizando a metodologia de varia??o do sinal fotoel?trico foi determinada a temperatura de cristaliza??o dos sistemas e avaliadas as interfer?ncias dos aditivos quanto ? redu??o da TIAC. Os resultados s?o expressos em fun??o das varia??es do sinal fotoel?trico, durante resfriamento controlado, inovando e validando esta metodologia de determina??o da TIAC, relativamente simples com rela??o ?s outras aplicadas que envolvem equipamentos espec?ficos e de alto custo. Atrav?s das curvas diferencias dos resultados tamb?m foram identificadas ?s etapas cr?ticas de crescimento e aglomera??o dos cristais que representam ? satura??o do sistema, indicando dificuldades de fluxo devido ao aumento da densidade

Bovine tuberculosis is a global cause for concern in livestock, free-ranging wildlife, zoological collections and the human population. Large amount of time, effort and resources are spent on its diagnosis and control methods. This study was aimed at determining the sensitivity and specificity of the IS6110 specific PCR test on formalin fixed, paraffin embedded (FFPE) tissue blocks, compared to that of the gold standard method culture and to differentiate M. bovis from other members of the M. tuberculosis complex using the RD4 region of difference specific PCR test. A total of 141 FFPE tissue blocks of wild animals from game reserves, the National Zoological Gardens and routine tuberculosis (TB) surveys in Kruger National Park were tested. Among the 50 known TB positive samples (35 M. bovis culture positive, twelve M. tuberculosis culture positive and three diagnosed tuberculosis positive on histopathology examination) the IS6110 PCR had an overall sensitivity of 22%. The positive predictive value of the IS6110 test (91.67%) was quite high implying that although sensitivity was low, one can be highly confident that a positive test result is a true reflection of the positive disease status. The overall sensitivity of the RD4 PCR was 20%. The positive predictive value of the RD4 test (41.67%) was low, implying that a positive test result may be unreliable. The sensitivities of the M. tuberculosis and M. bovis culture positive samples were compared and a significant difference was noted. Sensitivities of the IS6110 and RD4 assays in M. tuberculosis culture positive samples were 66.67% and 33.33%, respectively; sensitivities of the IS6110 and RD4 assays in M. bovis culture positive samples were 8.57% and 17.14%, respectively. Difference in bacterial load in tissues infected with the two mycobacterial species may account for this finding (i.e. M. bovis infections have a lower bacteria load). Of the 91 known TB negative samples, the specificity of the IS6110 (98.90%) and RD4 (84.62%) PCR tests were high, but the negative predictive values of 69.67% and 65.81%, respectively, suggest that the probability of negative test results being incorrect still exists. The resultant sensitivity was increased when parallel interpretation was applied to histopathology examination and the IS6110 or RD4 PCR tests and when applied to the IS6110 and RD4 PCR tests. Both histopathology examination and PCR tests produce rapid results and their combination can be used in routine diagnostics. The RD4 PCR assay was unable to distinguish M. bovis from other members of the MTB complex and based on the findings of this study the RD4 PCR cannot add value to the diagnosis of suspect tuberculosis samples at this stage, but successful troubleshooting relating to 1) extraction method, 2) DNA inhibitors, 3) contamination and 4) multisampling protocol, may enable its use in future. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Veterinary Tropical Diseases / Unrestricted

Philosophiae Doctor - PhD / Tumour-specific protein markers are usually present at elevated concentrations in patient biopsy tissue; therefore tumour tissue is an ideal biological material for studying cancer proteomics and biomarker discovery studies. To understand and elucidate cancer pathogenesis and its mechanisms at the molecular level, the collection and characterisation of a large number of individual patient tissue cohorts are required. Since most pathology institutes routinely preserve biopsy tissues by standardised methods of formalin fixation and paraffin embedment, these archived, FFPE tissues are important collections of pathology material, often accompanied by important metadata, such as patient medical history and treatments. FFPE tissue blocks are conveniently stored under ambient conditions for decades, while retaining cellular morphology due to the modifications induced by formalin. / 2022