Recuperação e purificação do antígeno 503 de Leishmania i. chagasi expresso em E. coli e remoção de endotoxina utilizando adsorção em leito expandido

The growing interest and applications of biotechnology products have increased the development of new processes for recovery and purification of proteins. The expanded bed adsorption (EBA) has emerged as a promising technique for this purpose. It combines into one operation the steps of clarification, concentration and purification of the target molecule. Hence, the method reduces the time and the cost of operation. In this context, this thesis aim was to evaluate the recovery and purification of 503 antigen of Leishmania i. chagasi expressed in E. coli M15 and endotoxin removal by EBA. In the first step of this study, batch experiments were carried out using two experimental designs to define the optimal adsorption and elution conditions of 503 antigen onto Streamline chelating resin. For adsorption assays, using expanded bed, it was used a column of 2.6 cm in diameter by 30.0 cm in height coupled to a peristaltic pump. In the second step of study, the removal of endotoxin during antigen recovery process was evaluated employing the non-ionic surfactant Triton X-114 in the washing step ALE. In the third step, we sought developing a mathematical model able to predict the 503 antigen breakthrough curves in expanded mode. The experimental design results to adsorption showed the pH 8.0 and the NaCl concentration of 2.4 M as the optimum adsorption condition. In the second design, the only significant factor for elution was the concentration of imidazole, which was taken at 600 mM. The adsorption isotherm of the 503 antigen showed a good fit to the Langmuir model (R = 0.98) and values for qmax (maximum adsorption capacity) and Kd (equilibrium constant) estimated were 1.95 mg/g and 0.34 mg/mL, respectively. Purification tests directly from unclarified feedstock showed a recovery of 59.2% of the target protein and a purification factor of 6.0. The addition of the non-ionic surfactant Triton X-114 to the washing step of EBA led to high levels (> 99%) of LPS removal initially present in the samples for all conditions tested. The mathematical model obtained to describe the 503 antigen breakthrough curves in Streamline Chelanting resin in expanded mode showed a good fit for both parameter estimation and validation steps. The validated model was used to optimize the efficiencies, achieving maximum values of the process and of the column efficiencies of 89.2% and 75.9%, respectively. Therefore, EBA is an efficient alternative for the recovery of the target protein and removal of endotoxin from an E. coli unclarified feedstock in just one step.

Análise da estrutura e transporte do zooplâncton no estuário do Rio Caravelas (Bahia-Brasil) através do uso do zooscan

Spatial-temporal dynamics of zooplankton in the Caravelas river estuary (Bahia, Brazil). The survey was conducted in order to describe the zooplankton community of the estuary Caravelas (Bahia, Brazil), to quantify and relate the patterns of horizontal and vertical transport with the type of tide (neap and spring) and tidal phase (flood and ebb). Zooplankton samples were collected with the aid of a suction pump (300L), filtered in plankton nets (300μm) and fixed in saline formalin 4%. Samples were collected at a fixed point (A1), near the mouth of the estuary, with samples taken at neap tides and spring tides during the dry and rainy seasons. Samples were collected for 13 hours, at intervals of 1 hour in 3 depths: surface, middle and bottom. Simultaneous collection of biological, we measured the current velocity, temperature and salinity of the water through CTD. In the laboratory, samples were selected for analysis in estereomicroscope, with 25 groups identified, with Copepoda getting the highest number of species. The 168 samples obtained from temporal samples were subsampled and processed on equipment ZooScan, with the aid of software ZooProcess at the end were generated 458.997 vingnettes. 8 taxa were identified automatically, with 16 classified as a semi-automatic. The group Copepoda, despite the limited taxonomic refinement ZooScan, obtained 2 genera and 1 species identified automatically. Among the seasons dry and wet groups Brachyura (zoea), Chaetognatha, and the Calanoid copepods (others), Temora spp., Oithona spp. and Euterpina acutifrons were those who had higher frequency of occurrence, appearing in more than 70% of the samples. Copepoda group showed the largest percentage of relative abundance in both seasons. There was no seasonal variation of total zooplankton, with an average density of 7826±4219 org.m-3 in the dry season, and 7959±3675 org.m-3 in the rainy season, neither between the types and phases of the tides, but seasonal differences were significant recorded for the main zooplankton groups. Vertical stratification was seen for the major zooplankton groups (Brachyura, Chaetognatha, Calanoida (other), Oithona spp, Temora spp. e Euterpina acutifrons). The scale of this stratification varied with the type (square or tide) and tidal phase (flood or ebb). The instantaneous transport was more influenced by current velocity, with higher values observed in spring tides to the total zooplankton, however, there was a variation of this pattern depending on the zooplankton group. According to the data import and export of total zooplankton, the outflow of organisms of the estuary was higher than the input. The results suggest that the estuary of Caravelas may influence the dynamics of organic matter to the adjacent coast, with possible consequences in National Marine Park of Abrolhos

Sistema de sensoriamento eletromagnético utilizado para detecção da contaminação do óleo isolante do motor no método de elevação artificial do tipo bombeio centrífugo submerso

In the artificial lift method by Electrical Submersible Pump (ESP), the energy is transmitted for the well´s deep through a flat electric handle, where it is converted into mechanical energy through an engine of sub-surface, which is connected to a centrifugal pump. This transmits energy to the fluid under the pressure form, bringing it to the surface In this method the subsurface equipment is basically divided into: pump, seal and motor. The main function of the seal is the protect the motor, avoiding the motor´s oil be contaminated by oil production and the consequent burning of it. Over time, the seal will be wearing and initiates a contamination of motor oil, causing it to lose its insulating characteristics. This work presents a design of a magnetic sensor capable of detecting contamination of insulating oil used in the artificial lift method of oil-type Electrical Submersible Pump (ESP). The objective of this sensor is to generate alarm signal just the moment when the contamination in the isolated oil is present, enabling the implementation of a predictive maintenance. The prototype was designed to work in harsh conditions to reach a depth of 2000m and temperatures up to 150°C. It was used a simulator software to defined the mechanical and electromagnetic variables. Results of field experiments were performed to validate the prototype. The final results performed in an ESP system with a 62HP motor showed a good reliability and fast response of the prototype.

Controle aplicado a poços com método de elevação e bombeio centrífugo submerso

The Electrical Submersible Pumping is an artificial lift method for oil wells employed in onshore and offshore areas. The economic revenue of the petroleum production in a well depends on the oil flow and the availability of lifting equipment. The fewer the failures, the lower the revenue shortfall and costs to repair it. The frequency with which failures occur depends on the operating conditions to which the pumps are submitted. In high-productivity offshore wells monitoring is done by operators with engineering support 24h/day, which is not economically viable for the land areas. In this context, the automation of onshore wells has clear economic advantages. This work proposes a system capable of automatically control the operation of electrical submersible pumps, installed in oil wells, by an adjustment at the electric motor rotation based on signals provided by sensors installed on the surface and subsurface, keeping the pump operating within the recommended range, closest to the well s potential. Techniques are developed to estimate unmeasured variables, enabling the automation of wells that do not have all the required sensors. The automatic adjustment, according to an algorithm that runs on a programmable logic controller maintains the flow and submergence within acceptable parameters avoiding undesirable operating conditions, as the gas interference and high engine temperature, without need to resort to stopping the engine, which would reduce the its useful life. The control strategy described, based on modeling of physical phenomena and operational experience reported in literature, is materialized in terms of a fuzzy controller based on rules, and all generated information can be accompanied by a supervisory system