Performance data of the coral Pocillopora verrucosa and environmental conditions along the Saudi Arabian Red Sea coast as assessed during 3 expeditions in the years 2011 and 2012

Large scale patterns of ecologically relevant traits may help identify drivers of their variability and conditions beneficial or adverse to the expression of these traits. Antimicrofouling defenses in scleractinian corals regulate the establishment of the associated biofilm as well as the risks of infection. The Saudi Arabian Red Sea coast features a pronounced thermal and nutritional gradient including regions and seasons with potentially stressful conditions to corals. Assessing the patterns of antimicrofouling defenses across the Red Sea may hint at the susceptibility of corals to global change. We investigated microfouling pressure as well as the relative strength of 2 alternative antimicrofouling defenses (chemical antisettlement activity, mucus release) along the pronounced environmental gradient along the Saudi Arabian Red Sea coast in 2 successive years. Microfouling pressure was exceptionally low along most of the coast but sharply increased at the southernmost sites. Mucus release correlated with temperature. Chemical defense tended to anti-correlate with mucus release. As a result, the combined action of mucus release and chemical antimicrofouling defense seemed to warrant sufficient defense against microbes along the entire coast. In the future, however, we expect enhanced energetic strain on corals when warming and/or eutrophication lead to higher bacterial fouling pressure and a shift towards putatively more costly defense by mucus release.

Survey and brain storming studies about machines, constructions, human and environmental risk consideration in the careers of the Universidad Politécnica of Madrid

The Universidad Politécnica of Madrid (UPM) includes schools and faculties that were for engineering degrees, architecture and computer science, that are now in a quick EEES Bolonia Plan metamorphosis getting into degrees, masters and doctorate structures. They are focused towards action in machines, constructions, enterprises, that are subjected to machines, human and environment created risks. These are present in actions such as use loads, wind, snow, waves, flows, earthquakes, forces and effects in machines, vehicles behavior, chemical effects, and other environmental factors including effects of crops, cattle and beasts, forests, and varied essential economic and social disturbances. Emphasis is for authors in this session more about risks of natural origin, such as for hail, winds, snow or waves that are not exactly known a priori, but that are often considered with statistical expected distributions giving extreme values for convenient return periods. These distributions are known from measures in time, statistic of extremes and models about hazard scenarios and about responses of man made constructions or devices. In each engineering field theories were built about hazards scenarios and how to cover for important risks. Engineers must get that the systems they handle, such as vehicles, machines, firms or agro lands or forests, obtain production with enough safety for persons and with decent economic results in spite of risks. For that risks must be considered in planning, in realization and in operation, and safety margins must be taken but at a reasonable cost. That is a small level of risks will often remain, due to limitations in costs or because of due to strange hazards, and maybe they will be covered by insurance in cases such as in transport with cars, ships or aircrafts, in agro for hail, or for fire in houses or in forests. These and other decisions about quality, security for men or about business financial risks are sometimes considered with Decision Theories models, using often tools from Statistics or operational Research. The authors have done and are following field surveys about risk consideration in the careers in UPM, making deep analysis of curricula taking into account the new structures of degrees in the EEES Bolonia Plan, and they have considered the risk structures offered by diverse schools of Decision theories. That gives an aspect of the needs and uses, and recommendations about improving in the teaching about risk, that may include special subjects especially oriented for each career, school or faculty, so as to be recommended to be included into the curricula, including an elaboration and presentation format using a multi-criteria decision model.

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.

Exploring new roles for the rpoS gene in the survival and virulence of the fire blight pathogen Erwinia amylovora

Erwinia amylovora causes fire blight in economically important plants of the family Rosaceae. This bacterial pathogen spends part of its life cycle coping with starvation and other fluctuating environmental conditions. In many Gram-negative bacteria, starvation and other stress responses are regulated by the sigma factor RpoS. We obtained an E. amylovora rpoS mutant to explore the role of this gene in starvation responses and its potential implication in other processes not yet studied in this pathogen. Results showed that E. amylovora needs rpoS to develop normal starvation survival and viable but nonculturable (VBNC) responses. Furthermore, this gene contributed to stationary phase cross-protection against oxidative, osmotic, and acid stresses and was essential for cross-protection against heat shock, but nonessential against acid shock. RpoS also mediated regulation of motility, exopolysaccharide synthesis, and virulence in immature loquats, but not in pear plantlets, and contributed to E. amylovora survival in nonhost tissues during incompatible interactions. Our results reveal some unique roles for the rpoS gene in E. amylovora and provide new knowledge on the regulation of different processes related to its ecology, including survival in different environments and virulence in immature fruits.

Paisajes de la sal en Andalucía

Los paisajes protagonistas de esta investigación han sido creados gracias a la actividad de las salinas, explotaciones “donde se beneficia la sal de las aguas del mar o de ciertos manantiales, cuando se ha evaporado el agua.” (RAE). Son lugares de características únicas, tanto por sus valores naturales y medioambientales, como por los valores culturales, patrimoniales, históricos, sociales e identitarios, “resultado de la acción e interacción de factores naturales y humanos en el territorio” (CEP, 2000). Existen salinas en muchos lugares del mundo, pero es en la Península Ibérica donde se encuentra la mayor concentración de estas explotaciones en toda Europa y, por consiguiente, de los singulares paisajes asociados y generados por ellas. Es también la única región europea donde existen salinas de interior que utilizan métodos de evaporación para la obtención de sal: las energías y circunstancias naturales aprovechadas en ellas, sol, viento, gravedad y humedad relativa, propician la evaporación del agua salada, una de las estrategias fundamentales para su funcionamiento. Esta circunstancia determina su posición geográfica en el territorio y su dependencia de los ciclos climáticos estacionales, del régimen hidrológico, la estructura geológica del subsuelo, la geomorfología y la acción humana para potenciar la eficiencia de los procesos productivos, creando para ello un extenso e interesante patrimonio natural y cultural en torno a ellas. Por tanto, son también paisajes culturales, resultado de un proceso artesanal de producción y comercialización de la sal, que se articula a través de la construcción de una serie de instalaciones preindustriales diseminadas y conectadas en puntos estratégicos del territorio costero e interior. La presente tesis aborda la grave situación a la que se enfrentan en la actualidad los paisajes ibéricos de la sal, concretamente los ubicados en la Comunidad Autónoma de Andalucía, que están sufriendo un ávido proceso de abandono y desaparición desde mediados del s. XX, con la consiguiente pérdida para la sociedad de este valioso patrimonio cultural y natural, que se ha ido construyendo durante siglos, prácticamente desde el inicio de la humanidad. De las diversas tipologías de salinas que existen, se han seleccionado las explotaciones, tanto marítimas como de interior, que utilizan o han utilizado en su origen técnicas de explotación artesanal basadas en la evaporación. El interés de esta acotación tipológica se ha basado en las relaciones de dependencia que estas explotaciones de sal establecen con el entorno físico y climático donde se ubican para poder existir y funcionar. El objetivo principal de esta investigación ha perseguido el establecimiento y definición de un marco territorial, global e integrador, para los paisajes culturales salineros andaluces, a través de la observación directa realizada durante el trabajo de campo, el análisis de textos, imágenes, documentos, gráficos, palabras y entrevistas abiertas. Lo anterior ha permitido identificar, clasificar, analizar y valorar sus principales características y su evolución en el tiempo, identificando las causas del deterioro y desaparición del patrimonio salinero, evaluando la efectividad de las iniciativas, públicas y/o privadas, y de las figuras de protección, así como analizar las relaciones que establecen con su entorno próximo, con los núcleos de población y habitantes a los que sirven, la red de carreteas y caminos que las conectan y los ríos y mares que las proveen de materia prima. Finalmente, se ha establecido una sistemática integral de análisis, con el fin de poder actuar, a partir del conocimiento, a favor de la gestión, salvaguarda y pervivencia de los excepcionales valores culturales y naturales, que definen el carácter e identidad de estos singulares paisajes. ----------------------ABSTRACT----------------- The landscapes of this research have been created by the activity of saltworks, sites “where salt from sea water or certain springs is obtained, when water evaporates.” (RAE). They are places with unique characteristics, because of their natural and environmental values, as well as their cultural, historic and social values, their heritage and identity, “the result of the action and interaction of natural and human factors on the territory” (European Landscape Convention, 2000). There are many saltworks in many places around the world, but it is in the Iberian Peninsula where we find the highest concentration of salt industries within Europe and, therefore, the particular landscapes associated and created by them. It is as well the only European region where we can find inland saltworks with evaporation methods to obtain salt: the energies and natural circumstances used in them are sun, wind, gravity and relative humidity, favouring salt water evaporation, one of the main strategies of their functioning. This circumstance determines their geographic position in the territory and its dependence on seasonal climatic cycles, hydrological regimes, geological subsoil structure, geomorphology and human action to strengthen productive processes efficiency, creating an extensive and interesting natural and cultural heritage around them. Therefore, they are also cultural landscapes, result of a traditional salt production process and marketing, organized through the construction of certain preindustrial buildings scattered and connected in strategic inland and seaside spots. This thesis deals with the plight today’s salt landscapes are facing, particularly those within the Andalucian Region, since the middle of the 20th century undergoing an avid process of abandonment and disappearance, meaning a great loss for society of a cultural and natural heritage constructed along centuries, almost since the beginning of mankind. Among the diverse typologies of saltworks, the selection made consists on those inland and seaside ones, which use or have originally used traditional production techniques based on evaporation. The interest in this particular saltwork typology is based on the dependency relationships these industries establish with the environment and climate where they are located, in order to be able to exist and function. The main goal of this research has pursued to establish and define a global and inclusive territorial framework for Andalucian cultural saltworks landscapes, through direct observation carried out during fieldwork, analysis of texts, images, documents, graphs, words and open interviews. All of it has allowed to identify, classify, analyze and evaluate their main characteristics and evolution over time, identifying the causes of deterioration and disappearance of the saltworks heritage, assessing the effectiveness of public and private initiatives, and protection projects, as well as analyzing the relationships with their surroundings, population centers and residents they serve, road networks connecting them and rivers and seas supplying the raw material. Finally, a comprehensive analysis systematic has been established, in order to be able to take action, with knowledge as starting point, for the management, preservation and survival of the unique cultural and natural values that define the character and identity of these singular landscapes.

New agri-environmental policies for the sustainability of mountainous olive farming. Evidences from Andalusia, Spain

La sostenibilidad de los sistemas olivareros situados en zonas de pendiente y montaña (SMOPS) en Andalucía se encuentra actualmente amenazada por las elevadas tasas de abandono que afectan a estos sistemas productivos. Así, la consumación de este proceso de abandono, no sólo pondría en peligro a las propias explotaciones, sino a todo el conjunto de bienes y servicios no productivos y al patrimonio cultural generado por este sistema productivo. En este contexto, la búsqueda de alternativas políticas enfocadas a revertir este proceso se erige como una necesidad categórica en aras de garantizar en el largo plazo la sostenibilidad de los olivares de montaña. Esta tesis pretende hacer frente a esta necesidad a través de la construcción de un marco político alternativo para los SMOPS, que permita la integración simultánea de todas las dimensiones que pueden influir en su desarrollo; esto es: el marco político actual, principalmente determinado por la Política Agraria Común (PAC) de la Unión Europea (UE); las preferencias de la sociedad hacia la oferta de bienes y servicios públicos generados por los SMOPS; y las preferencias y voluntad de innovación hacia nuevos manejos y sistemas de gestión de los agricultores y propietarios de las explotaciones. Para ello, se emplea una metodología de investigación mixta, que abarca la realización de cuatro encuestas (personales y online) llevadas a cabo a los agentes o grupos de interés involucrados directa o indirectamente en la gestión de los SMOPS –ciudadanos, agricultores y propietarios y expertos-; una profunda revisión de las herramientas de política agroambiental actuales y posibles alternativas a las mismas; y el desarrollo de nuevas estrategias metodológicas para dotar de mayor precisión y fiabilidad las estimaciones obtenidas a partir del Método del Experimento de Elección (MEE) en el campo de la valoración medioambiental. En general, los resultados muestran que una estrategia de política agroambiental basada en la combinación de los Contratos Territoriales de Zona Rural (CTZR) y el manejo ecológico supondría una mejora en la sostenibilidad de los sistemas olivareros de montaña andaluces, que, al mismo tiempo, propiciaría una mejor consideración de las necesidades y demandas de los agentes implicados en su gestión. Asimismo, los hallazgos obtenidos en esta investigación demandan un cambio de paradigma en los actuales pagos agroambientales, que han de pasar de una estrategia basada en la implementación de acciones, a otra enfocada al logro de objetivos, la cual, en el caso del olivar, se podría centrar en el aumento del secuestro de carbono en el suelo. Desde un punto de vista metodológico, los resultados han contribuido notablemente a mejorar la fiabilidad y precisión de las conclusiones estimadas a partir del MEE, mediante el diseño de un novedoso proceso iterativo para detectar posibles comportamientos inconsistentes por parte de los entrevistados con respecto a su máxima Disposición al Pago (DAP) para lograr la situación considerada como “óptima” en los olivares ecológicos de montaña andaluces. En líneas generales, el actual marco institucional favorece la puesta en práctica de la mayoría de las estrategias propuestas en esta tesis; sin embargo son necesarios mayores esfuerzos para reconducir los actuales Pagos Agroambientales y Climáticos de la PAC, hacia una estrategia de política agroambiental adaptada a las necesidades y requisitos del territorio en el que se aplica, enfocada al logro de objetivos y que sea capaz de integrar y coordinar al conjunto de agentes y grupos de interés involucrados -directa o indirectamente- en la gestión de los olivares de montaña. En este contexto, se espera que la puesta en práctica de nuevas estructuras y acuerdos de gobernanza territorial juegue un importante papel en el desarrollo de una política agroambiental realmente adaptada a las necesidades de los sistemas olivareros de montaña andaluces. ABSTRACT The long-term sustainability of Andalusian sloping and mountainous olive production systems (SMOPS) is currently threatened by the high abandonment rates that affect these production systems. The effective occurrence of this abandonment process is indeed menacing not only farms themselves, but also the wide array of public goods and services provided by SMOPS and the cultural heritage held by this production system. The search of policy alternatives aimed at tackling this process is thus a central necessity. This thesis aims to undertake this necessity by building an alternative policy framework for SMOPS that simultaneously integrates the several dimensions that are susceptible to influence it, namely: the current policy framework, mainly determined by the European Union’s (EU) Common Agricultural Policy (CAP); the social preferences toward the supply of SMOPS’ public goods and services; and farmers’ preferences and willingness to innovate toward new management practices in their farms. For this purpose, we put into practice a mixed-method strategy that combines four face-to-face and online surveys carried out with SMOPS’ stakeholders -including citizens, farmers and experts-; in-depth analysis of current and alternative agrienvironmental policy (AEP) instruments; and the development of novel methodological approaches to advance toward more reliable Discrete Choice Experiment’s (DCE) outcomes in the field of environmental valuation. Overall, results show that a policy strategy based on the combination of Territorial Management Contracts (TMC) and organic management would further enhance Andalusian SMOPS’ sustainability by simultaneously taking into account stakeholders’ demands and needs. Findings also call for paradigm shift of the current action-oriented design of Agri-Environmental-Climate Schemes (AECS), toward a result-based approach, that in the case of olive orchards should particularly be focused on enhancing soil carbon sequestration. From a methodological perspective, results have contributed to improve the accuracy and feasibility of DCE outcomes by designing a novel and iterative procedure focused in ascertaining respondents’ inconsistent behaviour with respect to their stated maximum WTP for the attainment of an ideal situation to be achieved in organic Andalusian SMOPS. Generally, the present institutional framework favours the implementation of the main policy strategies proposed in this thesis, albeit further efforts are required to better conduct current CAP’s agri-environmental instruments toward a territorially targeted result-oriented strategy capable to integrate and coordinate the whole set of stakeholders involved in the management of SMOPS. In this regard, alternative governance structures and arrangements are expected to play a major role on the process of tackling SMOPS’ agri-environmental policy challenge.

Paisajes de la sal en Andalucía

Los paisajes protagonistas de esta investigación han sido creados gracias a la actividad de las salinas, explotaciones “donde se beneficia la sal de las aguas del mar o de ciertos manantiales, cuando se ha evaporado el agua.” (RAE). Son lugares de características únicas, tanto por sus valores naturales y medioambientales, como por los valores culturales, patrimoniales, históricos, sociales e identitarios, “resultado de la acción e interacción de factores naturales y humanos en el territorio” (CEP, 2000). Existen salinas en muchos lugares del mundo, pero es en la Península Ibérica donde se encuentra la mayor concentración de estas explotaciones en toda Europa y, por consiguiente, de los singulares paisajes asociados y generados por ellas. Es también la única región europea donde existen salinas de interior que utilizan métodos de evaporación para la obtención de sal: las energías y circunstancias naturales aprovechadas en ellas, sol, viento, gravedad y humedad relativa, propician la evaporación del agua salada, una de las estrategias fundamentales para su funcionamiento. Esta circunstancia determina su posición geográfica en el territorio y su dependencia de los ciclos climáticos estacionales, del régimen hidrológico, la estructura geológica del subsuelo, la geomorfología y la acción humana para potenciar la eficiencia de los procesos productivos, creando para ello un extenso e interesante patrimonio natural y cultural en torno a ellas. Por tanto, son también paisajes culturales, resultado de un proceso artesanal de producción y comercialización de la sal, que se articula a través de la construcción de una serie de instalaciones preindustriales diseminadas y conectadas en puntos estratégicos del territorio costero e interior. La presente tesis aborda la grave situación a la que se enfrentan en la actualidad los paisajes ibéricos de la sal, concretamente los ubicados en la Comunidad Autónoma de Andalucía, que están sufriendo un ávido proceso de abandono y desaparición desde mediados del s. XX, con la consiguiente pérdida para la sociedad de este valioso patrimonio cultural y natural, que se ha ido construyendo durante siglos, prácticamente desde el inicio de la humanidad. De las diversas tipologías de salinas que existen, se han seleccionado las explotaciones, tanto marítimas como de interior, que utilizan o han utilizado en su origen técnicas de explotación artesanal basadas en la evaporación. El interés de esta acotación tipológica se ha basado en las relaciones de dependencia que estas explotaciones de sal establecen con el entorno físico y climático donde se ubican para poder existir y funcionar. El objetivo principal de esta investigación ha perseguido el establecimiento y definición de un marco territorial, global e integrador, para los paisajes culturales salineros andaluces, a través de la observación directa realizada durante el trabajo de campo, el análisis de textos, imágenes, documentos, gráficos, palabras y entrevistas abiertas. Lo anterior ha permitido identificar, clasificar, analizar y valorar sus principales características y su evolución en el tiempo, identificando las causas del deterioro y desaparición del patrimonio salinero, evaluando la efectividad de las iniciativas, públicas y/o privadas, y de las figuras de protección, así como analizar las relaciones que establecen con su entorno próximo, con los núcleos de población y habitantes a los que sirven, la red de carreteas y caminos que las conectan y los ríos y mares que las proveen de materia prima. Finalmente, se ha establecido una sistemática integral de análisis, con el fin de poder actuar, a partir del conocimiento, a favor de la gestión, salvaguarda y pervivencia de los excepcionales valores culturales y naturales, que definen el carácter e identidad de estos singulares paisajes. ----------------------ABSTRACT----------------- The landscapes of this research have been created by the activity of saltworks, sites “where salt from sea water or certain springs is obtained, when water evaporates.” (RAE). They are places with unique characteristics, because of their natural and environmental values, as well as their cultural, historic and social values, their heritage and identity, “the result of the action and interaction of natural and human factors on the territory” (European Landscape Convention, 2000). There are many saltworks in many places around the world, but it is in the Iberian Peninsula where we find the highest concentration of salt industries within Europe and, therefore, the particular landscapes associated and created by them. It is as well the only European region where we can find inland saltworks with evaporation methods to obtain salt: the energies and natural circumstances used in them are sun, wind, gravity and relative humidity, favouring salt water evaporation, one of the main strategies of their functioning. This circumstance determines their geographic position in the territory and its dependence on seasonal climatic cycles, hydrological regimes, geological subsoil structure, geomorphology and human action to strengthen productive processes efficiency, creating an extensive and interesting natural and cultural heritage around them. Therefore, they are also cultural landscapes, result of a traditional salt production process and marketing, organized through the construction of certain preindustrial buildings scattered and connected in strategic inland and seaside spots. This thesis deals with the plight today’s salt landscapes are facing, particularly those within the Andalucian Region, since the middle of the 20th century undergoing an avid process of abandonment and disappearance, meaning a great loss for society of a cultural and natural heritage constructed along centuries, almost since the beginning of mankind. Among the diverse typologies of saltworks, the selection made consists on those inland and seaside ones, which use or have originally used traditional production techniques based on evaporation. The interest in this particular saltwork typology is based on the dependency relationships these industries establish with the environment and climate where they are located, in order to be able to exist and function. The main goal of this research has pursued to establish and define a global and inclusive territorial framework for Andalucian cultural saltworks landscapes, through direct observation carried out during fieldwork, analysis of texts, images, documents, graphs, words and open interviews. All of it has allowed to identify, classify, analyze and evaluate their main characteristics and evolution over time, identifying the causes of deterioration and disappearance of the saltworks heritage, assessing the effectiveness of public and private initiatives, and protection projects, as well as analyzing the relationships with their surroundings, population centers and residents they serve, road networks connecting them and rivers and seas supplying the raw material. Finally, a comprehensive analysis systematic has been established, in order to be able to take action, with knowledge as starting point, for the management, preservation and survival of the unique cultural and natural values that define the character and identity of these singular landscapes.

Mode of action of the COR15a gene on the freezing tolerance of Arabidopsis thaliana

Constitutive expression of the cold-regulated COR15a gene of Arabidopsis thaliana results in a significant increase in the survival of isolated protoplasts frozen over the range of −4.5 to −7°C. The increased freezing tolerance is the result of a decreased incidence of freeze-induced lamellar-to-hexagonal II phase transitions that occur in regions where the plasma membrane is brought into close apposition with the chloroplast envelope as a result of freeze-induced dehydration. Moreover, the mature polypeptide encoded by this gene, COR15am, increases the lamellar-to-hexagonal II phase transition temperature of dioleoylphosphatidylethanolamine and promotes formation of the lamellar phase in a lipid mixture composed of the major lipid species that comprise the chloroplast envelope. We propose that COR15am, which is located in the chloroplast stroma, defers freeze-induced formation of the hexagonal II phase to lower temperatures (lower hydrations) by altering the intrinsic curvature of the inner membrane of the chloroplast envelope.

VanX, a bacterial d-alanyl-d-alanine dipeptidase: Resistance, immunity, or survival function?

The zinc-containing d-alanyl-d-alanine (d-Ala-d-Ala) dipeptidase VanX has been detected in both Gram-positive and Gram-negative bacteria, where it appears to have adapted to at least three distinct physiological roles. In pathogenic vancomycin-resistant enterococci, vanX is part of a five-gene cluster that is switched on to reprogram cell-wall biosynthesis to produce peptidoglycan chain precursors terminating in d-alanyl-d-lactate (d-Ala-d-lactate) rather than d-Ala-d-Ala. The modified peptidoglycan exhibits a 1,000-fold decrease in affinity for vancomycin, accounting for the observed phenotypic resistance. In the glycopeptide antibiotic producers Streptomyces toyocaensis and Amylocatopsis orientalis, a vanHAX operon may have coevolved with antibiotic biosynthesis genes to provide immunity by reprogramming cell-wall termini to d-Ala-d-lactate as antibiotic biosynthesis is initiated. In the Gram-negative bacterium Escherichia coli, which is never challenged by the glycopeptide antibiotics because they cannot penetrate the outer membrane permeability barrier, the vanX homologue (ddpX) is cotranscribed with a putative dipeptide transport system (ddpABCDF) in stationary phase by the transcription factor RpoS (σs). The combined action of DdpX and the permease would permit hydrolysis of d-Ala-d-Ala transported back into the cytoplasm from the periplasm as cell-wall crosslinks are refashioned. The d-Ala product could then be oxidized as an energy source for cell survival under starvation conditions.

Action spectra for phytochrome A- and B-specific photoinduction of seed germination in Arabidopsis thaliana.

We have examined the seed germination in Arabidopsis thaliana of wild type (wt), and phytochrome A (PhyA)- and B (PhyB)-mutants in terms of incubation time and environmental light effects. Seed germination of the wt and PhyA-null mutant (phyA) was photoreversibly regulated by red and far-red lights of 10-1,000 micromol m-2 when incubated in darkness for 1-14 hr, but no germination occurred in PhyB-null mutant (phyB). When wt seeds and the phyB mutant seeds were incubated in darkness for 48 hr, they synthesized PhyA during dark incubation and germinated upon exposure to red light of 1-100 nmol m-2 and far-red light of 0.5-10 micromol m-2, whereas the phyA mutant showed no such response. The results indicate that the seed germination is regulated by PhyA and PhyB but not by other phytochromes, and the effects of PhyA and PhyB are separable in this assay. We determined action spectra separately for PhyA- and PhyB-specific induction of seed germination at Okazaki large spectrograph. Action spectra for the PhyA response show that monochromatic 300-780 nm lights of very low fluence induced the germination, and this induction was not photoreversible in the range examined. Action spectra for the PhyB response show that germination was photoreversibly regulated by alternate irradiations with light of 0.01-1 mmol m-2 at wavelengths of 540-690 nm and 695-780 nm. The present work clearly demonstrated that PhyA photoirreversibly triggers the germination upon irradiations with ultraviolet, visible and far-red light of very low fluence, while PhyB controls the photoreversible effects of low fluence.

Jasmonic acid distribution and action in plants: regulation during development and response to biotic and abiotic stress.

Jasmonic acid (JA) is a naturally occurring growth regulator found in higher plants. Several physiological roles have been described for this compound (or a related compound, methyl jasmonate) during plant development and in response to biotic and abiotic stress. To accurately determine JA levels in plant tissue, we have synthesized JA containing 13C for use as an internal standard with an isotopic composition of [225]:[224] 0.98:0.02 compared with [225]:[224] 0.15:0.85 for natural material. GC analysis (flame ionization detection and MS) indicate that the internal standard is composed of 92% 2-(+/-)-[13C]JA and 8% 2-(+/-)-7-iso-[13C]JA. In soybean plants, JA levels were highest in young leaves, flowers, and fruit (highest in the pericarp). In soybean seeds and seedlings, JA levels were highest in the youngest organs including the hypocotyl hook, plumule, and 12-h axis. In soybean leaves that had been dehydrated to cause a 15% decrease in fresh weight, JA levels increased approximately 5-fold within 2 h and declined to approximately control levels by 4 h. In contrast, a lag time of 1-2 h occurred before abscisic acid accumulation reached a maximum. These results will be discussed in the context of multiple pathways for JA biosynthesis and the role of JA in plant development and responses to environmental signals.

Dominant-negative action of the jimpy mutation in mice complemented with an autosomal transgene for myelin proteolipid protein.

Mutations in genes encoding membrane proteins have been associated with cell death of unknown cause from invertebrate development to human degenerative diseases. A point mutation in the gene for myelin proteolipid protein (PLP) underlies oligodendrocyte death and dysmyelination in jimpy mice, an accurate model for Pelizaeus-Merzbacher disease. To distinguish the loss of PLP function from other effects of the misfolded protein, we took advantage of the X chromosomal linkage of the gene and have complemented jimpy with a wild-type PLP transgene. In this artificial heterozygous situation, the jimpy mutation emerged as genetically dominant. At the cellular level oligodendrocytes showed little increase in survival although endogenous PLP gene and autosomal transgene were truly coexpressed. In surviving oligodendrocytes, wild-type PLP was functional and immunodetectable in myelin. Moreover, compacted myelin sheaths regained their normal periodicity. This strongly suggests that, despite the presence of functional wild-type PLP, misfolded jimpy PLP is by itself the primary cause of abnormal oligodendrocyte death.

Intermediate hosts of Toxoplasma gondii and food safety:epidemiology, genetics and parasite survival in meat-producing animals in Italy

Toxoplasma gondii is a coccidian parasite with a global distribution. The definitive host is the cat (and other felids). All warm-blooded animals can act as intermediate hosts, including humans. Sexual reproduction (gametogony) takes place in the final host and oocysts are released in the environment, where they then sporulate to become infective. In intermediate hosts the cycle is extra-intestinal and results in the formation of tachyzoites and bradyzoites. Tachyzoites represent the invasive and proliferative stage and on entering a cell it multiplies asexually by endodyogeny. Bradyzoites within tissue cysts are the latent form. T. gondii is a food-borne parasite causing toxoplasmosis, which can occur in both animals and humans. Infection in humans is asymptomatic in more than 80% of cases in Europe and North-America. In the remaining cases patients present fever, cervical lymphadenopathy and other non-specific clinical signs. Nevertheless, toxoplasmosis is life threatening if it occurs in immunocompromised subjects. The main organs involved are brain (toxoplasmic encephalitis), heart (myocarditis), lungs (pulmonary toxoplasmosis), eyes, pancreas and parasite can be isolated from these tissues. Another aspect is congenital toxoplasmosis that may occur in pregnant women and the severity of the consequences depends on the stage of pregnancy when maternal infection occurs. Acute toxoplasmosis in developing foetuses may result in blindness, deformation, mental retardation or even death. The European Food Safety Authority (EFSA), in recent reports on zoonoses, highlighted that an increasing numbers of animals resulted infected with T. gondii in EU (reported by the European Member States for pigs, sheep, goats, hunted wild boar and hunted deer, in 2011 and 2012). In addition, high prevalence values have been detected in cats, cattle and dogs, as well as several other animal species, indicating the wide distribution of the parasite among different animal and wildlife species. The main route of transmission is consumption of food and water contaminated with sporulated oocysts. However, infection through the ingestion of meat contaminated with tissue cysts is frequent. Finally, although less frequent, other food products contaminated with tachyzoites such as milk, may also pose a risk. The importance of this parasite as a risk for human health was recently highlighted by EFSA’s opinion on modernization of meat inspection, where Toxoplasma gondii was identified as a relevant hazard to be addressed in revised meat inspection systems for pigs, sheep, goats, farmed wild boar and farmed deer (Call for proposals -GP/EFSA/BIOHAZ/2013/01). The risk of infection is more highly associated to animals reared outside, also in free-range or organic farms, where biohazard measure are less strict than in large scale, industrial farms. Here, animals are kept under strict biosecurity measures, including barriers, which inhibit access by cats, thus making soil contamination by oocysts nearly impossible. A growing demand by the consumer for organic products, coming from free-range livestock, in respect of animal-welfare, and the desire for the best quality of derived products, have all led to an increase in the farming of free-range animals. The risk of Toxoplasma gondii infection increases when animals have access to environment and the absence of data in Italy, together with need for in depth study of both the prevalence and genotypes of Toxoplasma gondii present in our country were the main reasons for the development of this thesis project. A total of 152 animals have been analyzed, including 21 free-range pigs (Suino Nero race), 24 transhumant Cornigliese sheep, 77 free-range chickens and 21 wild animals. Serology (on meat juice) and identification of T. gondii DNA through PCR was performed on all samples, except for wild animals (no serology). An in-vitro test was also applied with the aim to find an alternative and valid method to bioassay, actually the gold standard. Meat samples were digested and seeded onto Vero cells, checked every day and a RT-PCR protocol was used to determine an eventual increase in the amount of DNA, demonstrating the viability of the parasite. Several samples were alos genetically characterized using a PCR-RFLP protocol to define the major genotypes diffused in the geographical area studied. Within the context of a project promoted by Istituto Zooprofilattico of Pavia and Brescia (Italy), experimentally infected pigs were also analyzed. One of the aims was to verify if the production process of cured “Prosciutto di Parma” is able to kill the parasite. Our contribution included the digestion and seeding of homogenates on Vero cells and applying the Elisa test on meat juice. This thesis project has highlighted widespread diffusion of T. gondii in the geographical area taken into account. Pigs, sheep, chickens and wild animals showed high prevalence of infection. The data obtained with serology were 95.2%, 70.8%, 36.4%, respectively, indicating the spread of the parasite among numerous animal species. For wild animals, the average value of parasite infection determined through PCR was 44.8%. Meat juice serology appears to be a very useful, rapid and sensitive method for screening carcasses at slaughterhouse and for marketing “Toxo-free” meat. The results obtained on fresh pork meat (derived from experimentally infected pigs) before (on serum) and after (on meat juice) slaughter showed a good concordance. The free-range farming put in evidence a marked risk for meat-producing animals and as a consequence also for the consumer. Genotyping revealed the diffusion of Type-II and in a lower percentage of Type-III. In pigs is predominant the Type-II profile, while in wildlife is more diffused a Type-III and mixed profiles (mainly Type-II/III). The mixed genotypes (Type-II/III) could be explained by the presence of mixed infections. Free-range farming and the contact with wildlife could facilitate the spread of the parasite and the generation of new and atypical strains, with unknown consequences on human health. The curing process employed in this study appears to produce hams that do not pose a serious concern to human health and therefore could be marketed and consumed without significant health risk. Little is known about the diffusion and genotypes of T. gondii in wild animals; further studies on the way in which new and mixed genotypes may be introduced into the domestic cycle should be very interesting, also with the use of NGS techniques, more rapid and sensitive than PCR-RFLP. Furthermore wildlife can become a valuable indicator of environmental contamination with T. gondii oocysts. Other future perspectives regarding pigs include the expansion of the number of free-range animals and farms and for Cornigliese sheep the evaluation of other food products as raw milk and cheeses. It should be interesting to proceed with the validation of an ELISA test for infection in chickens, using both serum and meat juice on a larger number of animals and the same should be done also for wildlife (at the moment no ELISA tests are available and MAT is the reference method for them). Results related to Parma ham do not suggest a concerning risk for consumers. However, further studies are needed to complete the risk assessment and the analysis of other products cured using technological processes other than those investigated in the present study. For example, it could be interesting to analyze products such as salami, produced with pig meat all over the Italian country, with very different recipes, also in domestic and rural contexts, characterized by a very short period of curing (1 to 6 months). Toxoplasma gondii is one of the most diffuse food-borne parasites globally. Public health safety, improved animal production and protection of endangered livestock species are all important goals of research into reliable diagnostic tools for this infection. Future studies into the epidemiology, parasite survival and genotypes of T. gondii in meat producing animals should continue to be a research priority.

Discursive constructions of social and environmental accounting in Brazil: the case of Petrobras.

In the 29 years following \"Our Common Future\" by the United Nations, there is considerable debate among governments, civil society, interest groups and business organisations about what constitutes sustainable development, which constitutes evidence for a contested discourse concerning sustainability. The purpose of this study is to understand this debate in the developing economic context of Brazil, and in particular, to understand and critique the social and environmental accounting [SEA] discursive constructions relating to the State-owned, Petrobras as well as to understand the Brazilian literature on SEA. The discourse theory [DT]-based analysis employs rhetorical redescription to analyse twenty-two reports from Petrobras from 2004-2013. I investigate the political notions by employing the methodological framework of the Logics of Critical Explanation [LCE]. LCE engenders five methodological steps: problematisation, retroduction, logics (social, political and fantasmatic), articulation and critique. The empirical discussion suggests that the hegemony of economic development operates to obfuscate, rhetorically, the development of sustainability, so as to maintain the core business of Petrobras conceived as capital accumulation. Equally, these articulations also illustrate how the constructions of SEA operate to serve the company\'s purpose with few (none) profound changes in integration of sustainability. The Brazilian literature on SEA sustains the status quo of neo-liberal market policies that operate to protect the dominant business case approach to maintain an agenda of wealth-creation in relation to social and environmental needs. The articulations of the case manifested in policies regarding, for example, corruption, which involved over-payments for contracts and unsustainable practices relating to the use of fossil fuels and demonstrated that there was antagonism between action and disclosure. The corruption scandal that emerged after SEA disclosures highlighted the rhetorical nature of disclosure when financial resources were subtracted from the company for political parties and engineering contractors hid facts through incomplete disclosures. The articulations of SEA misrepresent a broader context of the meanings associated with sustainability, which restricted the constructions of SEA to principally serve and represent the intention of the most powerful groups. The significance of SEA, then is narrowed to represent particular interests. The study argues for more critical studies as limited Brazilian literature concerning SEA kept a \'safe distance\' from substantively critiquing the constructions of SEA and its articulations in the Brazilian context. The literature review and the Petrobras\' case illustrate a variety of naming, instituting and articulatory practices that endeavour to maintain the current hegemony of development in an emerging economy, which allows Petrobras to continue to exercise significant profit at the expense of the social and environmental. The constructed idea of development in Petrobras\' discourses emphasises a rhetoric of wider development, but, in reality, these discourses were the antithesis of political, social and ethical developmental issues. These constructions aim to hide struggles between social inequalities and exploitation of natural resources and constitute excuses about a fanciful notion of rhetorical and hegemonic neo-liberal development. In summary, this thesis contributes to the prior literature in five ways: (i) the addition of DT to the analysis of SEA enhances the discussion of political elements such as hegemony, antagonism, logic of equivalence/difference, ideology and articulation; (ii) the analysis of an emerging economy such as Brazil incorporates a new perspective of the discussion of the discourses of SEA and development; (iii) this thesis includes a focus on rhetoric to discuss the maintenance of the status quo; (iv) the holistic structure of the LCE approach enlarges the understanding of the social, political and fantasmatic logics of SEA studies and; (v) this thesis combines an analysis of the literature and the case of Petrobras to characterise and critique the state of the Brazilian academy and its impacts and reflections on the significance of SEA. This thesis, therefore, argues for more critical studies in the Brazilian academy due to the persistence of idea of SEA and development that takes-for-granted deep exclusions and contradictions and provide little space for critiques.

Coastal Erosion--Causes, Erosion Control Methods, Economic and Environmental Impacts, and Best Management Practices

Coastal erosion is an important and constant issue facing coastal areas all over the world today. The rate of coastal development over the years has increased, in turn requiring that action be taken to protect structures from the threat of erosion. A review of the causes of coastal erosion and the methods implemented to control it was conducted in order to determine the best course of action in response to coastal erosion issues. The potential positive and negative economic and environmental impacts are key concerns in determining whether or not to restore an eroding beach and which erosion control method(s) to implement. Results focus on providing a comparison of these concerns as well as recommendations for addressing coastal erosion issues.