Problem-based learning supported by semantic techniques

Problem-based learning has been applied over the last three decades to a diverse range of learning environments. In this educational approach, different problems are posed to the learners so that they can develop different solutions while learning about the problem domain. When applied to conceptual modelling, and particularly to Qualitative Reasoning, the solutions to problems are models that represent the behaviour of a dynamic system. The learner?s task then is to bridge the gap between their initial model, as their first attempt to represent the system, and the target models that provide solutions to that problem. We propose the use of semantic technologies and resources to help in bridging that gap by providing links to terminology and formal definitions, and matching techniques to allow learners to benefit from existing models.

Ultraintense lasers applied to laser fusion material testing: production of ions, X rays and neutrons

Due to the particular characteristics of the fusion products, i.e. very short pulses (less than a few μs long for ions when arriving to the walls; less than 1 ns long for X-rays), very high fluences ( 10 13 particles/cm 2 for both ions and X rays photons) and broad particle energy spectra (up to 10 MeV ions and 100 keV photons), the laser fusion community lacks of facilities to accurately test plasma facing materials under those conditions. In the present work, the ability of ultraintese lasers to create short pulses of energetic particles and high fluences is addressed as a solution to reproduce those ion and X-ray bursts. Based on those parameters, a comparison between fusion ion and laser driven ion beams is presented and discussed, describing a possible experimental set-up to generate with lasers the appropriate ion pulses. At the same time, the possibility of generating X-ray or neutron beams which simulate those of laser fusion environments is also indicated and assessed under current laser intensities. It is concluded that ultraintense lasers should play a relevant role in the validation of materials for laser fusion facilities.

Robust regression applied to fractal/multifractal analysis.

Fractal and multifractal are concepts that have grown increasingly popular in recent years in the soil analysis, along with the development of fractal models. One of the common steps is to calculate the slope of a linear fit commonly using least squares method. This shouldn?t be a special problem, however, in many situations using experimental data the researcher has to select the range of scales at which is going to work neglecting the rest of points to achieve the best linearity that in this type of analysis is necessary. Robust regression is a form of regression analysis designed to circumvent some limitations of traditional parametric and non-parametric methods. In this method we don?t have to assume that the outlier point is simply an extreme observation drawn from the tail of a normal distribution not compromising the validity of the regression results. In this work we have evaluated the capacity of robust regression to select the points in the experimental data used trying to avoid subjective choices. Based on this analysis we have developed a new work methodology that implies two basic steps: ? Evaluation of the improvement of linear fitting when consecutive points are eliminated based on R pvalue. In this way we consider the implications of reducing the number of points. ? Evaluation of the significance of slope difference between fitting with the two extremes points and fitted with the available points. We compare the results applying this methodology and the common used least squares one. The data selected for these comparisons are coming from experimental soil roughness transect and simulated based on middle point displacement method adding tendencies and noise. The results are discussed indicating the advantages and disadvantages of each methodology.

E-MIS validity applied to TELMA enhanced learning environment

Enhanced learning environments are arising with great success within the field of cognitive skills training in minimally invasive surgery (MIS) because they provides multiple benefits since they avoid time, spatial and cost constraints. TELMA [1,2] is a new technology enhanced learning platform that promotes collaborative and ubiquitous training of surgeons. This platform is based on four main modules: an authoring tool, a learning content and knowledge management system, an evaluation module and a professional network. TELMA has been designed and developed focused on the user; therefore it is necessary to carry out a user validation as final stage of the development. For this purpose, e-MIS validity [3] has been defined. This validation includes usability, contents and functionality validities both for the development and production stages of any e-Learning web platform. Using e-MIS validity, the e-Learning is fully validated since it includes subjective and objective metrics. The purpose of this study is to specify and apply a set of objective and subjective metrics using e-MIS validity to test usability, contents and functionality of TELMA environment within the development stage.

New Intermediate band sulphide nanoparticles acting in the full visible light range spectra as an active photocatalyst

Nowadays one of the challenges of materials science is to find new technologies that will be able to make the most of renewable energies. An example of new proposals in this field are the intermediate-band (IB) materials, which promise higher efficiencies in photovoltaic applications (through the intermediate band solar cells), or in heterogeneous photocatalysis (using nanoparticles of them, for the light-induced degradation of pollutants or for the efficient photoevolution of hydrogen from water). An IB material consists in a semiconductor in which gap a new level is introduced [1], the intermediate band (IB), which should be partially filled by electrons and completely separated of the valence band (VB) and of the conduction band (CB). This scheme (figure 1) allows an electron from the VB to be promoted to the IB, and from the latter to the CB, upon absorption of photons with energy below the band gap Eg, so that energy can be absorbed in a wider range of the solar spectrum and a higher current can be obtained without sacrificing the photovoltage (or the chemical driving force) corresponding to the full bandgap Eg, thus increasing the overall efficiency. This concept, applied to photocatalysis, would allow using photons of a wider visible range while keeping the same redox capacity. It is important to note that this concept differs from the classic photocatalyst doping principle, which essentially tries just to decrease the bandgap. This new type of materials would keep the full bandgap potential but would use also lower energy photons. In our group several IB materials have been proposed, mainly for the photovoltaic application, based on extensively doping known semiconductors with transition metals [2], examining with DFT calculations their electronic structures. Here we refer to In2S3 and SnS2, which contain octahedral cations; when doped with Ti or V an IB is formed according to quantum calculations (see e.g. figure 2). We have used a solvotermal synthesis method to prepare in nanocrystalline form the In2S3 thiospinel and the layered compound SnS2 (which when undoped have bandgaps of 2.0 and 2.2 eV respectively) where the cation is substituted by vanadium at a ?10% level. This substitution has been studied, characterizing the materials by different physical and chemical techniques (TXRF, XRD, HR-TEM/EDS) (see e.g. figure 3) and verifying with UV spectrometry that this substitution introduces in the spectrum the sub-bandgap features predicted by the calculations (figure 4). For both sulphide type nanoparticles (doped and undoped) the photocatalytic activity was studied by following at room temperature the oxidation of formic acid in aqueous suspension, a simple reaction which is easily monitored by UV-Vis spectroscopy. The spectral response of the process is measured using a collection of band pass filters that allow only some wavelengths into the reaction system. Thanks to this method the spectral range in which the materials are active in the photodecomposition (which coincides with the band gap for the undoped samples) can be checked, proving that for the vanadium substituted samples this range is increased, making possible to cover all the visible light range. Furthermore it is checked that these new materials are more photocorrosion resistant than the toxic CdS witch is a well know compound frequently used in tests of visible light photocatalysis. These materials are thus promising not only for degradation of pollutants (or for photovoltaic cells) but also for efficient photoevolution of hydrogen from water; work in this direction is now being pursued.

New control architecturebased on PXI for a 3-finger haptic device applied to virtual manipulation

To perform advanced manipulation of remote environments such as grasping, more than one finger is required implying higher requirements for the control architecture. This paper presents the design and control of a modular 3-finger haptic device that can be used to interact with virtual scenarios or to teleoperate dexterous remote hands. In a modular haptic device, each module allows the interaction with a scenario by using a single finger; hence, multi-finger interaction can be achieved by adding more modules. Control requirements for a multifinger haptic device are analyzed and new hardware/software architecture for these kinds of devices is proposed. The software architecture described in this paper is distributed and the different modules communicate to allow the remote manipulation. Moreover, an application in which this haptic device is used to interact with a virtual scenario is shown.

Survey on integration of Applied Geology and Geomorphology in Civil Engineering curricula in the framework of the European Higher Education Area (EHEA) by using practical trips

The engineer must have sufficient theoretical knowledge to be applied to solve specific problems, with the necessary capacity to simplify these approaches, and taking into account factors such as speed, simplicity, quality and economy. In Geology, its ultimate goal is the exploration of the history of the geological events through observation, deduction, reasoning and, in exceptional cases by the direct underground exploration or experimentation. Experimentation is very limited in Geology. Reproduction laboratory of certain phenomena or geological processes is difficult because both time and space become a large scale. For this reason, some Earth Sciences are in a nearly descriptive stage whereas others closest to the experimental, Geophysics and Geochemistry, have assimilated progress experienced by the physics and chemistry. Thus, Anglo-Saxon countries clearly separate Engineering Geology from Geological Engineering, i.e. Applied Geology to the Geological Engineering concepts. Although there is a big professional overlap, the first one corresponds to scientific approach, while the last one corresponds to a technological one. Applied Geology to Engineering could be defined as the Science and Applied Geology to the design, construction and performance of engineering infrastructures in and field geology discipline. There has been much discussion on the primacy of theory over practice. Today prevails the exaggeration of practice, but you get good workers and routine and mediocre teachers. This idea forgets too that teaching problem is a problem of right balance. The approach of the action lines on the European Higher Education Area (EHEA) framework provides for such balance. Applied Geology subject represents the first real contact with the physical environment with the practice profession and works. Besides, the situation of the topic in the first trace of Study Plans for many students implies the link to other subjects and topics of the career (tunnels, dams, groundwater, roads, etc). This work analyses in depth the justification of such practical trips. It shows the criteria and methods of planning and the result which manifests itself in pupils. Once practical trips experience developed, the objective work tries to know about results and changes on student’s motivation in learning perspective. This is done regardless of the outcome of their knowledge achievements assessed properly and they are not subject to such work. For this objective, it has been designed a survey about their motivation before and after trip. Survey was made by the Unidad Docente de Geología Aplicada of the Departamento de Ingeniería y Morfología del Terreno (Escuela Técnica Superior de Ingenieros de Caminos, Canales y Puertos, Universidad Politécnica de Madrid). It was completely anonymous. Its objective was to collect the opinion of the student as a key agent of learning and teaching of the subject. All the work takes place under new teaching/learning criteria approach at the European framework in Higher Education. The results are exceptionally good with 90% of student’s participation and with very high scores in a number of questions as the itineraries, teachers and visited places (range of 4.5 to 4.2 in a 5 points scale). The majority of students are very satisfied (average of 4.5 in a 5 points scale).

Maximum likelihood estimation of new state space models for operational modal analysis

The modal analysis of a structural system consists on computing its vibrational modes. The experimental way to estimate these modes requires to excite the system with a measured or known input and then to measure the system output at different points using sensors. Finally, system inputs and outputs are used to compute the modes of vibration. When the system refers to large structures like buildings or bridges, the tests have to be performed in situ, so it is not possible to measure system inputs such as wind, traffic, . . .Even if a known input is applied, the procedure is usually difficult and expensive, and there are still uncontrolled disturbances acting at the time of the test. These facts led to the idea of computing the modes of vibration using only the measured vibrations and regardless of the inputs that originated them, whether they are ambient vibrations (wind, earthquakes, . . . ) or operational loads (traffic, human loading, . . . ). This procedure is usually called Operational Modal Analysis (OMA), and in general consists on to fit a mathematical model to the measured data assuming the unobserved excitations are realizations of a stationary stochastic process (usually white noise processes). Then, the modes of vibration are computed from the estimated model. The first issue investigated in this thesis is the performance of the Expectation- Maximization (EM) algorithm for the maximum likelihood estimation of the state space model in the field of OMA. The algorithm is described in detail and it is analysed how to apply it to vibration data. After that, it is compared to another well known method, the Stochastic Subspace Identification algorithm. The maximum likelihood estimate enjoys some optimal properties from a statistical point of view what makes it very attractive in practice, but the most remarkable property of the EM algorithm is that it can be used to address a wide range of situations in OMA. In this work, three additional state space models are proposed and estimated using the EM algorithm: • The first model is proposed to estimate the modes of vibration when several tests are performed in the same structural system. Instead of analyse record by record and then compute averages, the EM algorithm is extended for the joint estimation of the proposed state space model using all the available data. • The second state space model is used to estimate the modes of vibration when the number of available sensors is lower than the number of points to be tested. In these cases it is usual to perform several tests changing the position of the sensors from one test to the following (multiple setups of sensors). Here, the proposed state space model and the EM algorithm are used to estimate the modal parameters taking into account the data of all setups. • And last, a state space model is proposed to estimate the modes of vibration in the presence of unmeasured inputs that cannot be modelled as white noise processes. In these cases, the frequency components of the inputs cannot be separated from the eigenfrequencies of the system, and spurious modes are obtained in the identification process. The idea is to measure the response of the structure corresponding to different inputs; then, it is assumed that the parameters common to all the data correspond to the structure (modes of vibration), and the parameters found in a specific test correspond to the input in that test. The problem is solved using the proposed state space model and the EM algorithm. Resumen El análisis modal de un sistema estructural consiste en calcular sus modos de vibración. Para estimar estos modos experimentalmente es preciso excitar el sistema con entradas conocidas y registrar las salidas del sistema en diferentes puntos por medio de sensores. Finalmente, los modos de vibración se calculan utilizando las entradas y salidas registradas. Cuando el sistema es una gran estructura como un puente o un edificio, los experimentos tienen que realizarse in situ, por lo que no es posible registrar entradas al sistema tales como viento, tráfico, . . . Incluso si se aplica una entrada conocida, el procedimiento suele ser complicado y caro, y todavía están presentes perturbaciones no controladas que excitan el sistema durante el test. Estos hechos han llevado a la idea de calcular los modos de vibración utilizando sólo las vibraciones registradas en la estructura y sin tener en cuenta las cargas que las originan, ya sean cargas ambientales (viento, terremotos, . . . ) o cargas de explotación (tráfico, cargas humanas, . . . ). Este procedimiento se conoce en la literatura especializada como Análisis Modal Operacional, y en general consiste en ajustar un modelo matemático a los datos registrados adoptando la hipótesis de que las excitaciones no conocidas son realizaciones de un proceso estocástico estacionario (generalmente ruido blanco). Posteriormente, los modos de vibración se calculan a partir del modelo estimado. El primer problema que se ha investigado en esta tesis es la utilización de máxima verosimilitud y el algoritmo EM (Expectation-Maximization) para la estimación del modelo espacio de los estados en el ámbito del Análisis Modal Operacional. El algoritmo se describe en detalle y también se analiza como aplicarlo cuando se dispone de datos de vibraciones de una estructura. A continuación se compara con otro método muy conocido, el método de los Subespacios. Los estimadores máximo verosímiles presentan una serie de propiedades que los hacen óptimos desde un punto de vista estadístico, pero la propiedad más destacable del algoritmo EM es que puede utilizarse para resolver un amplio abanico de situaciones que se presentan en el Análisis Modal Operacional. En este trabajo se proponen y estiman tres modelos en el espacio de los estados: • El primer modelo se utiliza para estimar los modos de vibración cuando se dispone de datos correspondientes a varios experimentos realizados en la misma estructura. En lugar de analizar registro a registro y calcular promedios, se utiliza algoritmo EM para la estimación conjunta del modelo propuesto utilizando todos los datos disponibles. • El segundo modelo en el espacio de los estados propuesto se utiliza para estimar los modos de vibración cuando el número de sensores disponibles es menor que vi Resumen el número de puntos que se quieren analizar en la estructura. En estos casos es usual realizar varios ensayos cambiando la posición de los sensores de un ensayo a otro (múltiples configuraciones de sensores). En este trabajo se utiliza el algoritmo EM para estimar los parámetros modales teniendo en cuenta los datos de todas las configuraciones. • Por último, se propone otro modelo en el espacio de los estados para estimar los modos de vibración en la presencia de entradas al sistema que no pueden modelarse como procesos estocásticos de ruido blanco. En estos casos, las frecuencias de las entradas no se pueden separar de las frecuencias del sistema y se obtienen modos espurios en la fase de identificación. La idea es registrar la respuesta de la estructura correspondiente a diferentes entradas; entonces se adopta la hipótesis de que los parámetros comunes a todos los registros corresponden a la estructura (modos de vibración), y los parámetros encontrados en un registro específico corresponden a la entrada en dicho ensayo. El problema se resuelve utilizando el modelo propuesto y el algoritmo EM.

Estudio y desarrollo de sensores de fibra óptica para detección de vibraciones en ductos ascendentes submarinos

En esta tesis doctoral se describe el trabajo de investigación enfocado al estudio y desarrollo de sensores de fibra óptica para la detección de presión, flujo y vibraciones en ductos ascendentes submarinos utilizados en la extracción y transporte de hidrocarburos, con el objetivo de aplicarlos en los campos de explotación de aguas profundas en el Golfo de México pertenecientes a la Industria Petrolera Mexicana. El trabajo se ha enfocado al estudio y desarrollo de sensores ópticos cuasi distribuidos y distribuidos. En especial se ha profundizado en el uso y aplicación de las redes de Bragg (FBG) y de reflectómetros ópticos en el dominio del tiempo sensible a la fase (φ-OTDR). Los sensores de fibra óptica son especialmente interesantes para estas aplicaciones por sus ventajosas características como su inmunidad a interferencias electromagnéticas, capacidad de multiplexado, fiabilidad para trabajar en ambientes hostiles, altas temperaturas, altas presiones, ambientes salino-corrosivos, etc. Además, la fibra óptica no solo es un medio sensor sino que puede usarse como medio de transmisión. Se ha realizado un estudio del estado del arte y las ventajas que presentan los sensores ópticos puntuales, cuasi-distribuidos y distribuidos con respecto a los sensores convencionales. Se han estudiado y descrito los interrogadores de redes de Bragg y se ha desarrollado un método de calibración útil para los interrogadores existentes en el mercado, consiguiendo incertidumbres en la medida de la longitud de onda menores de ± 88 nm e incertidumbres relativas (la mas interesante en el campo de los sensores) menores de ±3 pm. Centrándose en la aplicación de las redes de Bragg en la industria del petróleo, se ha realizado un estudio en detalle del comportamiento que presentan las FBGs en un amplio rango de temperaturas de -40 ºC a 500 oC. Como resultado de este estudio se han evaluado las diferencias en los coeficientes de temperatura en diversos tramos de mas mismas, así como para diferentes recubrimientos protectores. En especial se ha encontrado y evaluado las diferencias de los diferentes recubrimientos en el intervalo de temperaturas entre -40 ºC y 60 ºC. En el caso del intervalo de altas temperaturas, entre 100 ºC y 500 ºC, se ha medido y comprobado el cambio uniforme del coeficiente de temperatura en 1pm/ºC por cada 100 ºC de aumento de temperatura, en redes independientemente del fabricante de las mismas. Se ha aplicado las FBG a la medición de manera no intrusiva de la presión interna en una tubería y a la medición del caudal de un fluido en una tubería, por la medida de diferencia de presión entre dos puntos de la misma. Además se ha realizado un estudio de detección de vibraciones en tuberías con fluidos. Finalmente, se ha implementado un sistema de detección distribuida de vibraciones aplicable a la detección de intrusos en las proximidades de los ductos, mediante un φ-OTDR. En este sistema se ha estudiado el efecto negativo de la inestabilidad de modulación que limita la detección de vibraciones distribuidas, su sensibilidad y su alcance. ABSTRACT This thesis describes the research work focused for the study and development of on optical fiber sensors for detecting pressure, flow and vibration in subsea pipes used in the extraction and transportation of hydrocarbons, in order to apply them in deepwater fields in the Gulf of Mexico belonging to the Mexican oil industry. The work has focused on the study and development of optical sensors distributed and quasi distributed. Especially was done on the use and application of fiber Bragg grating (FBG) and optical reflectometers time domain phase sensitive (φ-OTDR). The optical fiber sensors especially are interesting for these applications for their advantageous characteristics such as immunity to electromagnetic interference, multiplexing capability, reliability to work in harsh environments, high temperatures, high pressures, corrosive saline environments, etc. Furthermore, the optical fiber is not only a sensor means it can be used as transmission medium. We have performed a study of the state of the art and the advantages offered by optical sensors point, quasi-distributed and distributed over conventional sensors. Have studied and described interrogators Bragg grating and has developed a calibration method for interrogators useful for the existing interrogators in the market, resulting uncertainty in the measurement of the wavelength of less than ± 0.17 nm and uncertainties (the more interesting in the field of sensors) less than ± 3 pm. Focusing on the application of the Bragg gratings in the oil industry, has been studied in detail the behavior of the FBGs in a wide range of temperatures from -40 °C to 500 oC. As a result of this study we have evaluated the difference in temperature coefficients over various sections of the same, as well as different protective coatings. In particular evaluated and found the differences coatings in the range of temperatures between -40 º C and 60 º C. For the high temperature range between 20 ° C and 500 ° C, has been measured and verified the uniform change of the temperature coefficient at 1pm / ° C for each 100 ° C increase in temperature, in networks regardless of manufacturer thereof. FBG is applied to the non-intrusive measurement of internal pressure in a pipeline and measuring flow of a fluid in a pipe, by measuring the pressure difference between two points thereof. Therefore, has also made a study of detecting vibrations in pipes with fluids. Finally, we have implemented a distributed sensing system vibration applied to intrusion detection in the vicinity of the pipelines, by φ-OTDR. In this system we have studied the negative effect of modulation instability limits the distributed vibration detection, sensitivity and scope.

Estrategias de monitorización de CO2 y otros gases en los estudios de análogos naturales

La Universidad Politécnica de Madrid (UPM) y la Università degli Studi di Firenze (UniFi), bajo la coordinación técnica de AMPHOS21, participan desde 2009 en el proyecto de investigación “Estrategias de Monitorización de CO2 y otros gases en el estudio de Análogos Naturales”, financiado por la Fundación Ciudad de la Energía (CIUDEN) en el marco del Proyecto Compostilla OXYCFB300 (http://www.compostillaproject.eu), del Programa “European Energy Program for Recovery - EEPR”. El objetivo principal del proyecto fue el desarrollo y puesta a punto de metodologías de monitorización superficiales para su aplicación en el seguimiento y control de los emplazamientos donde se realice el almacenamiento geológico de CO2, analizando técnicas que permitan detectar y cuantificar las posibles fugas de CO2 a la atmósfera. Los trabajos se realizaron tanto en análogos naturales (españoles e italianos) como en la Planta de Desarrollo Tecnológico de Almacenamiento de CO2 de Hontomín. Las técnicas analizadas se centran en la medición de gases y aguas superficiales (de escorrentía y manantiales). En cuanto a la medición de gases se analizó el flujo de CO2 que emana desde el suelo a la atmósfera y la aplicabilidad de trazadores naturales (como el radón) para la detección e identificación de las fugas de CO2. En cuanto al análisis químico de las aguas se analizaron los datos geoquímicos e isotópicos y los gases disueltos en las aguas de los alrededores de la PDT de Hontomín, con objeto de determinar qué parámetros son los más apropiados para la detección de una posible migración del CO2 inyectado, o de la salmuera, a los ambientes superficiales. Las medidas de flujo de CO2 se realizaron con la técnica de la cámara de acúmulo. A pesar de ser una técnica desarrollada y aplicada en diferentes ámbitos científicos se estimó necesario adaptar un protocolo de medida y de análisis de datos a las características específicas de los proyectos de captura y almacenamiento de CO2 (CAC). Donde los flujos de CO2 esperados son bajos y en caso de producirse una fuga habrá que detectar pequeñas variaciones en los valores flujo con un “ruido” en la señal alto, debido a actividad biológica en el suelo. La medida de flujo de CO2 mediante la técnica de la cámara de acúmulo se puede realizar sin limpiar la superficie donde se coloca la cámara o limpiando y esperando al reequilibrio del flujo después de la distorsión al sistema. Sin embargo, los resultados obtenidos después de limpiar y esperar muestran menor dispersión, lo que nos indica que este procedimiento es el mejor para la monitorización de los complejos de almacenamiento geológico de CO2. El protocolo de medida resultante, utilizado para la obtención de la línea base de flujo de CO2 en Hontomín, sigue los siguiente pasos: a) con una espátula se prepara el punto de medición limpiando y retirando el recubrimiento vegetal o la primera capa compacta de suelo, b) se espera un tiempo para la realización de la medida de flujo, facilitando el reequilibrio del flujo del gas tras la alteración provocada en el suelo y c) se realiza la medida de flujo de CO2. Una vez realizada la medición de flujo de CO2, y detectada si existen zonas de anomalías, se debe estimar la cantidad de CO2 que se está escapando a la atmósfera (emanación total), con el objetivo de cuantificar la posible fuga. Existen un amplio rango de metodologías para realizar dicha estimación, siendo necesario entender cuáles son las más apropiadas para obtener el valor más representativo del sistema. En esta tesis se comparan seis técnicas estadísticas: media aritmética, estimador insegado de la media (aplicando la función de Sichel), remuestreo con reemplazamiento (bootstrap), separación en diferentes poblaciones mediante métodos gráficos y métodos basados en criterios de máxima verosimilitud, y la simulación Gaussiana secuencial. Para este análisis se realizaron ocho campañas de muestreo, tanto en la Planta de Desarrollo Tecnológico de Hontomón como en análogos naturales (italianos y españoles). Los resultados muestran que la simulación Gaussiana secuencial suele ser el método más preciso para realizar el cálculo, sin embargo, existen ocasiones donde otros métodos son más apropiados. Como consecuencia, se desarrolla un procedimiento de actuación para seleccionar el método que proporcione el mejor estimador. Este procedimiento consiste, en primer lugar, en realizar un análisis variográfico. Si existe una autocorrelación entre los datos, modelizada mediante el variograma, la mejor técnica para calcular la emanación total y su intervalo de confianza es la simulación Gaussiana secuencial (sGs). Si los datos son independientes se debe comprobar la distribución muestral, aplicando la media aritmética o el estimador insesgado de la media (Sichel) para datos normales o lognormales respectivamente. Cuando los datos no son normales o corresponden a una mezcla de poblaciones la mejor técnica de estimación es la de remuestreo con reemplazamiento (bootstrap). Siguiendo este procedimiento el máximo valor del intervalo de confianza estuvo en el orden del ±20/25%, con la mayoría de valores comprendidos entre ±3,5% y ±8%. La identificación de las diferentes poblaciones muestrales en los datos de flujo de CO2 puede ayudar a interpretar los resultados obtenidos, toda vez que esta distribución se ve afectada por la presencia de varios procesos geoquímicos como, por ejemplo, una fuente geológica o biológica del CO2. Así pues, este análisis puede ser una herramienta útil en el programa de monitorización, donde el principal objetivo es demostrar que no hay fugas desde el reservorio a la atmósfera y, si ocurren, detectarlas y cuantificarlas. Los resultados obtenidos muestran que el mejor proceso para realizar la separación de poblaciones está basado en criterios de máxima verosimilitud. Los procedimientos gráficos, aunque existen pautas para realizarlos, tienen un cierto grado de subjetividad en la interpretación de manera que los resultados son menos reproducibles. Durante el desarrollo de la tesis se analizó, en análogos naturales, la relación existente entre el CO2 y los isótopos del radón (222Rn y 220Rn), detectándose en todas las zonas de emisión de CO2 una relación positiva entre los valores de concentración de 222Rn en aire del suelo y el flujo de CO2. Comparando la concentración de 220Rn con el flujo de CO2 la relación no es tan clara, mientras que en algunos casos aumenta en otros se detecta una disminución, hecho que parece estar relacionado con la profundidad de origen del radón. Estos resultados confirmarían la posible aplicación de los isótopos del radón como trazadores del origen de los gases y su aplicación en la detección de fugas. Con respecto a la determinación de la línea base de flujo CO2 en la PDT de Hontomín, se realizaron mediciones con la cámara de acúmulo en las proximidades de los sondeos petrolíferos, perforados en los ochenta y denominados H-1, H-2, H-3 y H-4, en la zona donde se instalarán el sondeo de inyección (H-I) y el de monitorización (H-A) y en las proximidades de la falla sur. Desde noviembre de 2009 a abril de 2011 se realizaron siete campañas de muestreo, adquiriéndose más de 4.000 registros de flujo de CO2 con los que se determinó la línea base y su variación estacional. Los valores obtenidos fueron bajos (valores medios entre 5 y 13 g•m-2•d-1), detectándose pocos valores anómalos, principalmente en las proximidades del sondeo H-2. Sin embargo, estos valores no se pudieron asociar a una fuente profunda del CO2 y seguramente estuvieran más relacionados con procesos biológicos, como la respiración del suelo. No se detectaron valores anómalos cerca del sistema de fracturación (falla Ubierna), toda vez que en esta zona los valores de flujo son tan bajos como en el resto de puntos de muestreo. En este sentido, los valores de flujo de CO2 aparentemente están controlados por la actividad biológica, corroborado al obtenerse los menores valores durante los meses de otoño-invierno e ir aumentando en los periodos cálidos. Se calcularon dos grupos de valores de referencia, el primer grupo (UCL50) es 5 g•m-2•d-1 en las zonas no aradas en los meses de otoño-invierno y 3,5 y 12 g•m-2•d-1 en primavera-verano para zonas aradas y no aradas, respectivamente. El segundo grupo (UCL99) corresponde a 26 g•m-2•d- 1 durante los meses de otoño-invierno en las zonas no aradas y 34 y 42 g•m-2•d-1 para los meses de primavera-verano en zonas aradas y no aradas, respectivamente. Flujos mayores a estos valores de referencia podrían ser indicativos de una posible fuga durante la inyección y posterior a la misma. Los primeros datos geoquímicos e isotópicos de las aguas superficiales (de escorrentía y de manantiales) en el área de Hontomín–Huermeces fueron analizados. Los datos sugieren que las aguas estudiadas están relacionadas con aguas meteóricas con un circuito hidrogeológico superficial, caracterizadas por valores de TDS relativamente bajos (menor a 800 mg/L) y una fácie hidrogeoquímica de Ca2+(Mg2+)-HCO3 −. Algunas aguas de manantiales se caracterizan por concentraciones elevadas de NO3 − (concentraciones de hasta 123 mg/l), lo que sugiere una contaminación antropogénica. Se obtuvieron concentraciones anómalas de of Cl−, SO4 2−, As, B y Ba en dos manantiales cercanos a los sondeos petrolíferos y en el rio Ubierna, estos componentes son probablemente indicadores de una posible mezcla entre los acuíferos profundos y superficiales. El estudio de los gases disueltos en las aguas también evidencia el circuito superficial de las aguas. Estando, por lo general, dominado por la componente atmosférica (N2, O2 y Ar). Sin embargo, en algunos casos el gas predominante fue el CO2 (con concentraciones que llegan al 63% v/v), aunque los valores isotópicos del carbono (<-17,7 ‰) muestran que lo más probable es que esté relacionado con un origen biológico. Los datos geoquímicos e isotópicos de las aguas superficiales obtenidos en la zona de Hontomín se pueden considerar como el valor de fondo con el que comparar durante la fase operacional, la clausura y posterior a la clausura. En este sentido, la composición de los elementos mayoritarios y traza, la composición isotópica del carbono del CO2 disuelto y del TDIC (Carbono inorgánico disuelto) y algunos elementos traza se pueden considerar como parámetros adecuados para detectar la migración del CO2 a los ambientes superficiales. ABSTRACT Since 2009, a group made up of Universidad Politécnica de Madrid (UPM; Spain) and Università degli Studi Firenze (UniFi; Italy) has been taking part in a joint project called “Strategies for Monitoring CO2 and other Gases in Natural analogues”. The group was coordinated by AMPHOS XXI, a private company established in Barcelona. The Project was financially supported by Fundación Ciudad de la Energía (CIUDEN; Spain) as a part of the EC-funded OXYCFB300 project (European Energy Program for Recovery -EEPR-; www.compostillaproject.eu). The main objectives of the project were aimed to develop and optimize analytical methodologies to be applied at the surface to Monitor and Verify the feasibility of geologically stored carbon dioxide. These techniques were oriented to detect and quantify possible CO2 leakages to the atmosphere. Several investigations were made in natural analogues from Spain and Italy and in the Tecnchnological Development Plant for CO2 injection al Hontomín (Burgos, Spain). The studying techniques were mainly focused on the measurements of diffuse soil gases and surface and shallow waters. The soil-gas measurements included the determination of CO2 flux and the application to natural trace gases (e.g. radon) that may help to detect any CO2 leakage. As far as the water chemistry is concerned, geochemical and isotopic data related to surface and spring waters and dissolved gases in the area of the PDT of Hontomín were analyzed to determine the most suitable parameters to trace the migration of the injected CO2 into the near-surface environments. The accumulation chamber method was used to measure the diffuse emission of CO2 at the soil-atmosphere interface. Although this technique has widely been applied in different scientific areas, it was considered of the utmost importance to adapt the optimum methodology for measuring the CO2 soil flux and estimating the total CO2 output to the specific features of the site where CO2 is to be stored shortly. During the pre-injection phase CO2 fluxes are expected to be relatively low where in the intra- and post-injection phases, if leakages are to be occurring, small variation in CO2 flux might be detected when the CO2 “noise” is overcoming the biological activity of the soil (soil respiration). CO2 flux measurements by the accumulation chamber method could be performed without vegetation clearance or after vegetation clearance. However, the results obtained after clearance show less dispersion and this suggests that this procedure appears to be more suitable for monitoring CO2 Storage sites. The measurement protocol, applied for the determination of the CO2 flux baseline at Hontomín, has included the following steps: a) cleaning and removal of both the vegetal cover and top 2 cm of soil, b) waiting to reduce flux perturbation due to the soil removal and c) measuring the CO2 flux. Once completing the CO2 flux measurements and detected whether there were anomalies zones, the total CO2 output was estimated to quantify the amount of CO2 released to the atmosphere in each of the studied areas. There is a wide range of methodologies for the estimation of the CO2 output, which were applied to understand which one was the most representative. In this study six statistical methods are presented: arithmetic mean, minimum variances unbiased estimator, bootstrap resample, partitioning of data into different populations with a graphical and a maximum likelihood procedures, and sequential Gaussian simulation. Eight campaigns were carried out in the Hontomín CO2 Storage Technology Development Plant and in natural CO2 analogues. The results show that sequential Gaussian simulation is the most accurate method to estimate the total CO2 output and the confidential interval. Nevertheless, a variety of statistic methods were also used. As a consequence, an application procedure for selecting the most realistic method was developed. The first step to estimate the total emanation rate was the variogram analysis. If the relation among the data can be explained with the variogram, the best technique to calculate the total CO2 output and its confidence interval is the sequential Gaussian simulation method (sGs). If the data are independent, their distribution is to be analyzed. For normal and log-normal distribution the proper methods are the arithmetic mean and minimum variances unbiased estimator, respectively. If the data are not normal (log-normal) or are a mixture of different populations the best approach is the bootstrap resampling. According to these steps, the maximum confidence interval was about ±20/25%, with most of values between ±3.5% and ±8%. Partitioning of CO2 flux data into different populations may help to interpret the data as their distribution can be affected by different geochemical processes, e.g. geological or biological sources of CO2. Consequently, it may be an important tool in a monitoring CCS program, where the main goal is to demonstrate that there are not leakages from the reservoir to the atmosphere and, if occurring, to be able to detect and quantify it. Results show that the partitioning of populations is better performed by maximum likelihood criteria, since graphical procedures have a degree of subjectivity in the interpretation and results may not be reproducible. The relationship between CO2 flux and radon isotopes (222Rn and 220Rn) was studied in natural analogues. In all emissions zones, a positive relation between 222Rn and CO2 was observed. However, the relationship between activity of 220Rn and CO2 flux is not clear. In some cases the 220Rn activity indeed increased with the CO2 flux in other measurements a decrease was recognized. We can speculate that this effect was possibly related to the route (deep or shallow) of the radon source. These results may confirm the possible use of the radon isotopes as tracers for the gas origin and their application in the detection of leakages. With respect to the CO2 flux baseline at the TDP of Hontomín, soil flux measurements in the vicinity of oil boreholes, drilled in the eighties and named H-1 to H-4, and injection and monitoring wells were performed using an accumulation chamber. Seven surveys were carried out from November 2009 to summer 2011. More than 4,000 measurements were used to determine the baseline flux of CO2 and its seasonal variations. The measured values were relatively low (from 5 to 13 g•m-2•day-1) and few outliers were identified, mainly located close to the H-2 oil well. Nevertheless, these values cannot be associated to a deep source of CO2, being more likely related to biological processes, i.e. soil respiration. No anomalies were recognized close to the deep fault system (Ubierna Fault) detected by geophysical investigations. There, the CO2 flux is indeed as low as other measurement stations. CO2 fluxes appear to be controlled by the biological activity since the lowest values were recorded during autumn-winter seasons and they tend to increase in warm periods. Two reference CO2 flux values (UCL50 of 5 g•m-2•d-1 for non-ploughed areas in autumn-winter seasons and 3.5 and 12 g•m-2•d-1 for in ploughed and non-ploughed areas, respectively, in spring-summer time, and UCL99 of 26 g•m-2•d-1 for autumn-winter in not-ploughed areas and 34 and 42 g•m-2•d-1 for spring-summer in ploughed and not-ploughed areas, respectively, were calculated. Fluxes higher than these reference values could be indicative of possible leakage during the operational and post-closure stages of the storage project. The first geochemical and isotopic data related to surface and spring waters and dissolved gases in the area of Hontomín–Huermeces (Burgos, Spain) are presented and discussed. The chemical and features of the spring waters suggest that they are related to a shallow hydrogeological system as the concentration of the Total Dissolved Solids approaches 800 mg/L with a Ca2+(Mg2+)-HCO3 − composition, similar to that of the surface waters. Some spring waters are characterized by relatively high concentrations of NO3 − (up to 123 mg/L), unequivocally suggesting an anthropogenic source. Anomalous concentrations of Cl−, SO4 2−, As, B and Ba were measured in two springs, discharging a few hundred meters from the oil wells, and in the Rio Ubierna. These contents are possibly indicative of mixing processes between deep and shallow aquifers. The chemistry of the dissolved gases also evidences the shallow circuits of the Hontomín– Huermeces, mainly characterized by an atmospheric source as highlighted by the contents of N2, O2, Ar and their relative ratios. Nevertheless, significant concentrations (up to 63% by vol.) of isotopically negative CO2 (<−17.7‰ V-PDB) were found in some water samples, likely related to a biogenic source. The geochemical and isotopic data of the surface and spring waters in the surroundings of Hontomín can be considered as background values when intra- and post-injection monitoring programs will be carried out. In this respect, main and minor solutes, the isotopic carbon of dissolved CO2 and TDIC (Total Dissolved Inorganic Carbon) and selected trace elements can be considered as useful parameters to trace the migration of the injected CO2 into near-surface environments.

Mejora de la eficacia de Penicillium oxalicum como agente de biocontrol en enfermedades de plantas hortícolas

Actualmente, la reducción de materias activas (UE) y la implantación de la nueva Directiva comunitaria 2009/128/ que establece el marco de actuación para conseguir un uso sostenible de los plaguicidas químicos y la preferencia de uso de métodos biológicos, físicos y otros no químicos, obliga a buscar métodos de control menos perjudiciales para el medio ambiente. El control biológico (CB) de enfermedades vegetales empleando agentes de control biológico (ACB) se percibe como una alternativa más segura y con menor impacto ambiental, bien solos o bien como parte de una estrategia de control integrado. El aislado 212 de Penicillium oxalicum (PO212) (ATCC 201888) fue aislado originalmente de la micoflora del suelo en España y ha demostrado ser un eficaz ACB frente a la marchitez vascular del tomate. Una vez identificado y caracterizado el ACB se inició el periodo de desarrollo del mismo poniendo a punto un método de producción en masa de sus conidias. Tras lo cual se inició el proceso de formulación del ACB deshidratando las conidias para su preservación durante un período de tiempo mayor mediante lecho fluido. Finalmente, se han desarrollado algunos formulados que contienen de forma individual diferentes aditivos que han alargado su viabilidad, estabilidad y facilitado su manejo y aplicación. Sin embargo, es necesario seguir trabajando en la mejora de su eficacia de biocontrol. El primer objetivo de esta Tesis se ha centrado en el estudio de la interacción ACB-patógeno-huésped que permita la actuación de P.oxalicum en diferentes patosistemas. Uno de los primeros puntos que se abordan dentro de este objetivo es el desarrollo de nuevas FORMULACIONES del ACB que incrementen su eficacia frente a la marchitez vascular del tomate. Las conidias formuladas de PO212 se obtuvieron por la adición conjunta de distintos aditivos (mojantes, adherentes o estabilizantes) en dos momentos diferentes del proceso de producción/secado: i) antes del proceso de producción (en la bolsa de fermentación) en el momento de la inoculación de las bolsas de fermentación con conidias de PO212 o ii) antes del secado en el momento de la resuspensión de las conidias tras su centrifugación. De las 22 nuevas formulaciones desarrolladas y evaluadas en plantas de tomate en ensayos en invernadero, seis de ellas (FOR22, FOR25, FOR32, FOR35, FOR36 y FOR37) mejoran significativamente (P=0,05) el control de la marchitez vascular del tomate con respecto al obtenido con las conidias secas de P.oxalicum sin aditivos (CSPO) o con el fungicida Bavistin. Los formulados que mejoran la eficacia de las conidias secas sin aditivos son aquellos que contienen como humectantes alginato sódico en fermentación, seguido de aquellos que contienen glicerol como estabilizante en fermentación, y metil celulosa y leche desnatada como adherentes antes del secado. Además, el control de la marchitez vascular del tomate por parte de los formulados de P. oxalicum está relacionado con la fecha de inicio de la enfermedad. Otra forma de continuar mejorando la eficacia de biocontrol es mejorar la materia activa mediante la SELECCIÓN DE NUEVAS CEPAS de P. oxalicum, las cuales podrían tener diferentes niveles de eficacia. De entre las 28 nuevas cepas de P. oxalicum ensayadas en cámara de cultivo, sólo el aislado PO15 muestra el mismo nivel de eficacia que PO212 (62-67% de control) frente a la marchitez vascular del tomate en casos de alta presión de enfermedad. Mientras que, en casos de baja presión de enfermedad todas las cepas de P. oxalicum y sus mezclas demuestran ser eficaces. Finalmente, se estudia ampliar el rango de actuación de este ACB a OTROS HUÉSPEDES Y OTROS PATÓGENOS Y DIFERENTES GRADOS DE VIRULENCIA. En ensayos de eficacia de P. oxalicum frente a aislados de diferente agresividad de Verticillium spp. y Fusarium oxysporum f. sp. lycopersici en plantas de tomate en cámaras de cultivo, se demuestra que la eficacia de PO212 está negativamente correlacionada con el nivel de enfermedad causada por F. oxysporum f. sp. lycopersici pero que no hay ningún efecto diferencial en la reducción de la incidencia ni de la gravedad según la virulencia de los aislados. Sin embargo, en los ensayos realizados con V. dahliae, PO212 causa una mayor reducción de la enfermedad en las plantas inoculadas con aislados de virulencia media. La eficacia de PO212 también era mayor frente a aislados de virulencia media alta de F. oxysporum f. sp. melonis y F. oxysporum f. sp. niveum, en plantas de melón y sandía, respectivamente. En ambos huéspedes se demuestra que la dosis óptima de aplicación del ACB es de 107 conidias de PO212 g-1 de suelo de semillero, aplicada 7 días antes del trasplante. Además, entre 2 y 4 nuevas aplicaciones de PO212 a la raíces de las plantas mediante un riego al terreno de asiento mejoran la eficacia de biocontrol. La eficacia de PO212 no se limita a hongos patógenos vasculares como los citados anteriormente, sino también a otros patógenos como: Phytophthora cactorum, Globodera pallida y G. rostochiensis. PO212 reduce significativamente los síntomas (50%) causados por P. cactorum en plantas de vivero de fresa, tras la aplicación del ACB por inmersión de las raíces antes de su trasplante al suelo de viveros comerciales. Por otra parte, la exposición de los quistes de Globodera pallida y G. rostochiensis (nematodos del quiste de la patata) a las conidias de P. oxalicum, en ensayos in vitro o en microcosmos de suelo, reduce significativamente la capacidad de eclosión de los huevos. Para G. pallida esta reducción es mayor cuando se emplean exudados de raíz de patata del cv. 'Monalisa', que exudados de raíz del cv. 'Desirée'. No hay una reducción significativa en la tasa de eclosión con exudados de raíz de tomate del cv. 'San Pedro'. Para G. rostochiensis la reducción en la tasa de eclosión de los huevos se obtiene con exudados de la raíz de patata del cv. 'Desirée'. El tratamiento con P. oxalicum reduce también significativamente el número de quistes de G. pallida en macetas. Con el fin de optimizar la aplicación práctica de P. oxalicum cepa 212 como tratamiento biológico del suelo, es esencial entender cómo el entorno físico influye en la capacidad de colonización, crecimiento y supervivencia del mismo, así como el posible riesgo que puede suponer su aplicación sobre el resto de los microorganismos del ecosistema. Por ello en este segundo objetivo de esta tesis se estudia la interacción del ACB con el medio ambiente en el cual se aplica. Dentro de este objetivo se evalúa la INFLUENCIA DE LA TEMPERATURA, DISPONIBILIDAD DE AGUA Y PROPIEDADES FÍSICO-QUÍMICAS DE LOS SUELOS (POROSIDAD, TEXTURA, DENSIDAD...) SOBRE LA SUPERVIVENCIA Y EL CRECIMIENTO DE PO212 en condiciones controladas elaborando modelos que permitan predecir el impacto de cada factor ambiental en la supervivencia y crecimiento de P. oxalicum y conocer su capacidad para crecer y sobrevivir en diferentes ambientes. En las muestras de suelo se cuantifica: i) la supervivencia de Penicillium spp. usando el recuento del número de unidades formadoras de colonias en un medio de cultivo semi-selectivo y ii) el crecimiento (biomasa) de PO212 mediante PCR en tiempo real. En los resultados obtenidos se demuestra que P. oxalicum crece y sobrevive mejor en condiciones de sequía independientemente de la temperatura y del tipo de suelo. Si comparamos tipos de suelo P. oxalicum crece y sobrevive en mayor medida en suelos areno-arcillosos con un bajo contenido en materia orgánica, un mayor pH y una menor disponibilidad de fósforo y nitrógeno. La supervivencia y el crecimiento de P. oxalicum se correlaciona de forma negativa con la disponibilidad de agua y de forma positiva con el contenido de materia orgánica. Sólo la supervivencia se correlaciona también positivamente con el pH. Por otro lado se realizan ensayos en suelos de huertos comerciales con diferentes propiedades físico-químicas y diferentes condiciones ambientales para ESTUDIAR EL ESTABLECIMIENTO, SUPERVIVENCIA Y DISPERSIÓN VERTICAL Y MOVILIDAD HORIZONTAL DE PO212. P. oxalicum 212 puede persistir y sobrevivir en esos suelos al menos un año después de su liberación pero a niveles similares a los de otras especies de Penicillium indígenas presentes en los mismos suelos naturales. Además, P. oxalicum 212 muestra una dispersión vertical y movilidad horizontal muy limitada en los diferentes tipos de suelo evaluados. La introducción de P. oxalicum en un ambiente natural no sólo implica su actuación sobre el microorganismo diana, el patógeno, si no también sobre otros microorganismos indígenas. Para EVALUAR EL EFECTO DE LA APLICACIÓN DE P. oxalicum SOBRE LAS POBLACIONES FÚNGICAS INDIGENAS PRESENTES EN EL SUELO de dos huertos comerciales, se analizan mediante electroforesis en gradiente desnaturalizante de poliacrilamida (DGGE) muestras de dichos suelos a dos profundidades (5 y 10 cm) y a cuatro fechas desde la aplicación de P. oxalicum 212 (0, 75, 180 y 365 días). El análisis de la DGGE muestra que las diferencias entre las poblaciones fúngicas se deben significativamente a la fecha de muestreo y son independientes del tratamiento aplicado y de la profundidad a la que se tomen las muestras. Luego, la aplicación del ACB no afecta a la población fúngica de los dos suelos analizados. El análisis de las secuencias de la DGGE confirma los resultados anteriores y permiten identificar la presencia del ACB en los suelos. La presencia de P. oxalicum en el suelo se encuentra especialmente relacionada con factores ambientales como la humedad. Por tanto, podemos concluir que Penicillium oxalicum cepa 212 puede considerarse un óptimo Agente de Control Biológico (ACB), puesto que es ecológicamente competitivo, eficaz para combatir un amplio espectro de enfermedades y no supone un riesgo para el resto de microorganismos fúngicos no diana presentes en el lugar de aplicación. ABSTRACT Currently, reduction of active (EU) and the implementation of the new EU Directive 2009/128 which establishing the framework for action to achieve the sustainable use of chemical pesticides and preference of use of biological, physical and other non-chemical methods, forces to look for control methods less harmful to the environment. Biological control (CB) of plant diseases using biological control agents (BCA) is perceived as a safer alternative and with less environmental impact, either alone or as part of an integrated control strategy. The isolate 212 of Penicillium oxalicum (PO212) (ATCC 201888) was originally isolated from the soil mycoflora in Spain. P. oxalicum is a promising biological control agent for Fusarium wilt and other tomato diseases. Once identified and characterized the BCA, was developed a mass production method of conidia by solid-state fermentation. After determined the process of obtaining a formulated product of the BCA by drying of product by fluid-bed drying, it enables the preservation of the inoculum over a long period of time. Finally, some formulations of dried P. oxalicum conidia have been developed which contain one different additive that have improved their viability, stability and facilitated its handling and application. However, further work is needed to improve biocontrol efficacy. The first objective of this thesis has focused on the study of the interaction BCA- pathogen-host, to allow P.oxalicum to work in different pathosystems. The first point to be addressed in this objective is the development of new FORMULATIONS of BCA which increase their effectiveness against vascular wilt of tomato. PO212 conidial formulations were obtained by the joint addition of various additives (wetting agents, adhesives or stabilizers) at two different points of the production-drying process: i) to substrate in the fermentation bags before the production process, and (ii) to conidial paste obtained after production but before drying. Of the 22 new formulations developed and evaluated in tomato plants in greenhouse tests, six of them (FOR22 , FOR25 , FOR32 , FOR35 , FOR36 and FOR3) improved significantly (P = 0.05) the biocontrol efficacy against tomato wilt with respect to that obtained with dried P.oxalicum conidia without additives (CSPO) or the fungicide Bavistin. The formulations that improve the efficiency of dried conidia without additives are those containing as humectants sodium alginate in the fermentation bags, followed by those containing glycerol as a stabilizer in the fermentation bags, and methylcellulose and skimmed milk as adherents before drying. Moreover, control of vascular wilt of tomatoes by PO212 conidial formulations is related to the date of disease onset. Another way to further improve the effectiveness of biocontrol is to improve the active substance by SELECTION OF NEW STRAINS of P. oxalicum, which may have different levels of effectiveness. Of the 28 new strains of P. oxalicum tested in a culture chamber, only PO15 isolate shows the same effectiveness that PO212 (62-67 % of control) against tomato vascular wilt in cases of high disease pressure. Whereas in cases of low disease pressure all strains of P. oxalicum and its mixtures effective. Finally, we study extend the range of action of this BCA TO OTHER GUESTS AND OTHER PATHOGENS AND DIFFERENT DEGREES OF VIRULENCE. In efficacy trials of P. oxalicum against isolates of different aggressiveness of Verticillium spp. and Fusarium oxysporum f. sp. lycopersici in tomato plants in growth chambers, shows that the efficiency of PO212 is negatively correlated with the level of disease caused by F. oxysporum f. sp. lycopersici. There is not differential effect in reducing the incidence or severity depending on the virulence of isolates. However, PO212 cause a greater reduction of disease in plants inoculated with virulent isolates media of V. dahlia. PO212 efficacy was also higher against isolates of high and average virulence of F. oxysporum f. sp. melonis and F. oxysporum f. sp. niveum in melon and watermelon plants, respectively. In both hosts the optimum dose of the BCA application is 107 conidia PO212 g-1 soil, applied on seedlings 7 days before transplantation into the field. Moreover, the reapplication of PO212 (2-4 times) to the roots by irrigation into the field improve efficiency of biocontrol. The efficacy of PO212 is not limited to vascular pathogens as those mentioned above, but also other pathogens such as Oomycetes (Phytophthora cactorum) and nematodes (Globodera pallida and G. rostochiensis). PO212 significantly reduces symptoms (50 %) caused by P. cactorum in strawberry nursery plants after application of BCA by dipping the roots before transplanting to soil in commercial nurseries. Moreover, the exposure of G. pallida and G. rostochiensis cysts to the conidia of P. oxalicum, in in vitro assays or in soil microcosms significantly reduces hatchability of eggs. The reduction in the rate of G. pallida juveniles hatching was greatest when root diffusates from the `Monalisa´ potato cultivar were used, followed by root diffusates from the `Désirée´ potato cultivar. However, no significant reduction in the rate of G. pallida juveniles hatching was found when root diffusates from the ‘San Pedro” tomato cultivar were used. For G. rostochiensis reduction in the juveniles hatching is obtained from the root diffusates 'Desirée' potato cultivar. Treatment with P. oxalicum also significantly reduces the number of cysts of G. pallida in pots. In order to optimize the practical application of P. oxalicum strain 212 as a biological soil treatment, it is essential to understand how the physical environment influences the BCA colonization, survival and growth, and the possible risk that can cause its application on other microorganisms in the ecosystem of performance. Therefore, the second objective of this thesis is the interaction of the BCA with the environment in which it is applied. Within this objective is evaluated the INFLUENCE OF TEMPERATURE, WATER AVAILABILITY AND PHYSICAL-CHEMICAL PROPERTIES OF SOILS (POROSITY, TEXTURE, DENSITY...) ON SURVIVAL AND GROWTH OF PO212 under controlled conditions to develop models for predicting the environmental impact of each factor on survival and growth of P. oxalicum and to know their ability to grow and survive in different environments. Two parameters are evaluated in the soil samples: i) the survival of Penicillium spp. by counting the number of colony forming units in semi-selective medium and ii) growth (biomass) of PO212 by real-time PCR. P. oxalicum grows and survives better in drought conditions regardless of temperature and soil type. P. oxalicum grows and survives more in sandy loam soils with low organic matter content, higher pH and lower availability of phosphorus and nitrogen. Survival and growth of P. oxalicum negatively correlates with the availability of water and positively with the organic content. Only survival also correlated positively with pH. Moreover, trials are carried out into commercial orchards soils with different physic-chemical properties and different environmental conditions TO STUDY THE ESTABLISHMENT, SURVIVAL, VERTICAL DISPERSION AND HORIZONTAL SPREAD OF PO212. P. oxalicum 212 can persist and survive at very low levels in soil one year after its release. The size of the PO212 population after its release into the tested natural soils is similar to that of indigenous Penicillium spp. Furthermore, the vertical dispersion and horizontal spread of PO212 is limited in different soil types. The introduction of P. oxalicum in a natural environment not only involves their action on the target organism, the pathogen, but also on other indigenous microorganisms. TO ASSESS THE EFFECT OF P. oxalicum APPLICATION ON SOIL INDIGENOUS FUNGAL COMMUNITIES in two commercial orchards, soil samples are analyzed by Denaturing Gradient Gel Electrophoresis polyacrylamide (DGGE). Samples are taken from soil at two depths (5 and 10 cm) and four dates from the application of P. oxalicum 212 (0, 75, 180 and 365 days). DGGE analysis shows that differences are observed between sampling dates and are independent of the treatment of P. oxalicum applied and the depth. BCA application does not affect the fungal population of the two soil analyzed. Sequence analysis of the DGGE bands confirms previous findings and to identify the presence of BCA on soils. The presence of P. oxalicum in soil is especially related to environmental factors such as humidity. Therefore, we conclude that the 212 of strain Penicillium oxalicum can be considered an optimum BCA, since it is environmentally competitive and effective against a broad spectrum of diseases and does not have any negative effect on soil non-target fungi communities.

Localization and Tracking in Energy-Constrained Networks with Dynamic Connectivity

Systems used for target localization, such as goods, individuals, or animals, commonly rely on operational means to meet the final application demands. However, what would happen if some means were powered up randomly by harvesting systems? And what if those devices not randomly powered had their duty cycles restricted? Under what conditions would such an operation be tolerable in localization services? What if the references provided by nodes in a tracking problem were distorted? Moreover, there is an underlying topic common to the previous questions regarding the transfer of conceptual models to reality in field tests: what challenges are faced upon deploying a localization network that integrates energy harvesting modules? The application scenario of the system studied is a traditional herding environment of semi domesticated reindeer (Rangifer tarandus tarandus) in northern Scandinavia. In these conditions, information on approximate locations of reindeer is as important as environmental preservation. Herders also need cost-effective devices capable of operating unattended in, sometimes, extreme weather conditions. The analyses developed are worthy not only for the specific application environment presented, but also because they may serve as an approach to performance of navigation systems in absence of reasonably accurate references like the ones of the Global Positioning System (GPS). A number of energy-harvesting solutions, like thermal and radio-frequency harvesting, do not commonly provide power beyond one milliwatt. When they do, battery buffers may be needed (as it happens with solar energy) which may raise costs and make systems more dependent on environmental temperatures. In general, given our problem, a harvesting system is needed that be capable of providing energy bursts of, at least, some milliwatts. Many works on localization problems assume that devices have certain capabilities to determine unknown locations based on range-based techniques or fingerprinting which cannot be assumed in the approach considered herein. The system presented is akin to range-free techniques, but goes to the extent of considering very low node densities: most range-free techniques are, therefore, not applicable. Animal localization, in particular, uses to be supported by accurate devices such as GPS collars which deplete batteries in, maximum, a few days. Such short-life solutions are not particularly desirable in the framework considered. In tracking, the challenge may times addressed aims at attaining high precision levels from complex reliable hardware and thorough processing techniques. One of the challenges in this Thesis is the use of equipment with just part of its facilities in permanent operation, which may yield high input noise levels in the form of distorted reference points. The solution presented integrates a kinetic harvesting module in some nodes which are expected to be a majority in the network. These modules are capable of providing power bursts of some milliwatts which suffice to meet node energy demands. The usage of harvesting modules in the aforementioned conditions makes the system less dependent on environmental temperatures as no batteries are used in nodes with harvesters--it may be also an advantage in economic terms. There is a second kind of nodes. They are battery powered (without kinetic energy harvesters), and are, therefore, dependent on temperature and battery replacements. In addition, their operation is constrained by duty cycles in order to extend node lifetime and, consequently, their autonomy. There is, in turn, a third type of nodes (hotspots) which can be static or mobile. They are also battery-powered, and are used to retrieve information from the network so that it is presented to users. The system operational chain starts at the kinetic-powered nodes broadcasting their own identifier. If an identifier is received at a battery-powered node, the latter stores it for its records. Later, as the recording node meets a hotspot, its full record of detections is transferred to the hotspot. Every detection registry comprises, at least, a node identifier and the position read from its GPS module by the battery-operated node previously to detection. The characteristics of the system presented make the aforementioned operation own certain particularities which are also studied. First, identifier transmissions are random as they depend on movements at kinetic modules--reindeer movements in our application. Not every movement suffices since it must overcome a certain energy threshold. Second, identifier transmissions may not be heard unless there is a battery-powered node in the surroundings. Third, battery-powered nodes do not poll continuously their GPS module, hence localization errors rise even more. Let's recall at this point that such behavior is tight to the aforementioned power saving policies to extend node lifetime. Last, some time is elapsed between the instant an identifier random transmission is detected and the moment the user is aware of such a detection: it takes some time to find a hotspot. Tracking is posed as a problem of a single kinetically-powered target and a population of battery-operated nodes with higher densities than before in localization. Since the latter provide their approximate positions as reference locations, the study is again focused on assessing the impact of such distorted references on performance. Unlike in localization, distance-estimation capabilities based on signal parameters are assumed in this problem. Three variants of the Kalman filter family are applied in this context: the regular Kalman filter, the alpha-beta filter, and the unscented Kalman filter. The study enclosed hereafter comprises both field tests and simulations. Field tests were used mainly to assess the challenges related to power supply and operation in extreme conditions as well as to model nodes and some aspects of their operation in the application scenario. These models are the basics of the simulations developed later. The overall system performance is analyzed according to three metrics: number of detections per kinetic node, accuracy, and latency. The links between these metrics and the operational conditions are also discussed and characterized statistically. Subsequently, such statistical characterization is used to forecast performance figures given specific operational parameters. In tracking, also studied via simulations, nonlinear relationships are found between accuracy and duty cycles and cluster sizes of battery-operated nodes. The solution presented may be more complex in terms of network structure than existing solutions based on GPS collars. However, its main gain lies on taking advantage of users' error tolerance to reduce costs and become more environmentally friendly by diminishing the potential amount of batteries that can be lost. Whether it is applicable or not depends ultimately on the conditions and requirements imposed by users' needs and operational environments, which is, as it has been explained, one of the topics of this Thesis.

Improving the low-wind performance of the AERMOD atmospheric dispersion model for predicting short-range impacts of livestock ammonia emissions.

Short-range impacts to sensitive ecosystems as a result of ammonia emitted by livestock farms are often assessed using atmospheric dispersion modelling systems such as AERMOD. These assessments evaluate mean annual atmospheric concentrations of ammonia and nitrogen deposition rates at the ecosystem location for comparison with ecosystem damage thresholds. However, predictions of mean annual atmospheric concentrations can be dominated by periods of stable night-time conditions, which can contribute significantly to mean concentrations. AERMOD has been demonstrated to overestimate concentrations in certain stable low-wind conditions and so the model could potentially overestimate the short-range impacts of livestock ammonia emissions. This paper tests several modifications to the parameterisation of AERMOD (v12345) that aim to improve model predictions in low-wind conditions. The modifications are first described and then are applied to three pig farm case studies in the USA, Denmark and Spain to assess whether the modifications improve long-term mean ammonia concentration predictions through improved model performance. For these three case studies, most of the modifications tested improved model performance as a result of reducing the long-term mean concentration predictions, with the largest effect for low- or ground-level sources (e.g. slurry lagoons or naturally ventilated housing).

Rule Representation in Distributed Environments with Accepting Networks of Splicing Processors.

This paper presents the model named Accepting Networks of Evolutionary Processors as NP-problem solver inspired in the biological DNA operations. A processor has a rules set, splicing rules in this model,an object multiset and a filters set. Rules can be applied in parallel since there exists a large number of copies of objects in the multiset. Processors can form a graph in order to solve a given problem. This paper shows the network configuration in order to solve the SAT problem using linear resources and time. A rule representation arquitecture in distributed environments can be easily implemented using these networks of processors, such as decision support systems, as shown in the paper.

Traffic sign detection and tracking using robust 3D analysis

In this paper we present an innovative technique to tackle the problem of automatic road sign detection and tracking using an on-board stereo camera. It involves a continuous 3D analysis of the road sign during the whole tracking process. Firstly, a color and appearance based model is applied to generate road sign candidates in both stereo images. A sparse disparity map between the left and right images is then created for each candidate by using contour-based and SURF-based matching in the far and short range, respectively. Once the map has been computed, the correspondences are back-projected to generate a cloud of 3D points, and the best-fit plane is computed through RANSAC, ensuring robustness to outliers. Temporal consistency is enforced by means of a Kalman filter, which exploits the intrinsic smoothness of the 3D camera motion in traffic environments. Additionally, the estimation of the plane allows to correct deformations due to perspective, thus easing further sign classification.