Mordenite from Spain and its application as pozzolana

In Spain, natural zeolites have been only found in the volcanic areas of Canary Islands (San Gil, M.M., 1959). However, several years ago the study of the South Eastern region of continental Spain (Cabo de Gata) revealed the presence of a deposit of mordenite being industrially commercialized as bentonite. The deposit San José-Los Escullos is located in the concession registered as Los Murcianos, managed by Bentonitas Especiales, S.A. (BENESA) who was extracting bentonite while ignoring the presence of the zeolite. The name San José-Los Escullos refers to the discovery of mordenite in this deposit. The deposit is located in a volcanic area, constituted by pyroxenic andesites, breccias, tuffs, pyroclasts with andesitic and dacitic composition, largely altered by the hydrothermal solutions. The recent discovery of this zeolite with mordenite content up to 97%, has changed the initial vision about that deposit, and new possible uses of this mineral in the pozzolanic cement industry are foreseen. This work intends to give preliminary data on the characterization of this zeolite according to X-ray diffraction (XRD), ICP analyses, and scanning electron microscopy (SEM), as well as to emphasize the pozzolanic properties of this material, using both chemical and mechanical tests.

Drivers in adopting adaptation strategies to face increased water scarcity in agriculture. A case study in Doñana (Spain) and Makueni (Kenya)

La agricultura es uno de los sectores más afectados por el cambio climático. A pesar de haber demostrado a lo largo de la historia una gran capacidad para adaptarse a nuevas situaciones, hoy en día la agricultura se enfrenta a nuevos retos tales como satisfacer un elevado crecimiento en la demanda de alimentos, desarrollar una agricultura sostenible con el medio ambiente y reducir las emisiones de gases de efecto invernadero. El potencial de adaptación debe ser definido en un contexto que incluya el comportamiento humano, ya que éste juega un papel decisivo en la implementación final de las medidas. Por este motivo, y para desarrollar correctamente políticas que busquen influir en el comportamiento de los agricultores para fomentar la adaptación a estas nuevas condiciones, es necesario entender previamente los procesos de toma de decisiones a nivel individual o de explotación, así como los efectos de los factores que determinan las barreras o motivaciones de la implementación de medidas. Esta Tesis doctoral trata de profundizar en el análisis de factores que influyen en la toma de decisiones de los agricultores para adoptar estrategias de adaptación al cambio climático. Este trabajo revisa la literatura actual y desarrolla un marco metodológico a nivel local y regional. Dos casos de estudio a nivel local (Doñana, España y Makueni, Kenia) han sido llevados a cabo con el fin de explorar el comportamiento de los agricultores hacia la adaptación. Estos casos de estudio representan regiones con notables diferencias en climatología, impactos del cambio climático, barreras para la adaptación y niveles de desarrollo e influencia de las instituciones públicas y privadas en la agricultura. Mientras el caso de estudio de Doñana representa un ejemplo de problemas asociados al uso y escasez del agua donde se espera que se agraven en el futuro, el caso de estudio de Makueni ejemplifica una zona fuertemente amenazada por las predicciones de cambio climático, donde adicionalmente la falta de infraestructura y la tecnología juegan un papel crucial para la implementación de la adaptación. El caso de estudio a nivel regional trata de generalizar en África el comportamiento de los agricultores sobre la implementación de medidas. El marco metodológico que se ha seguido en este trabajo abarca una amplia gama de enfoques y métodos para la recolección y análisis de datos. Los métodos utilizados para la toma de datos incluyen la implementación de encuestas, entrevistas, talleres con grupos de interés, grupos focales de discusión, revisión de estudios previos y bases de datos públicas. Los métodos analíticos incluyen métodos estadísticos, análisis multi‐criterio para la toma de decisiones, modelos de optimización de uso del suelo y un índice compuesto calculado a través de indicadores. Los métodos estadísticos se han utilizado con el fin de evaluar la influencia de los factores socio‐económicos y psicológicos sobre la adopción de medidas de adaptación. Dentro de estos métodos se incluyen regresiones logísticas, análisis de componentes principales y modelos de ecuaciones estructurales. Mientras que el análisis multi‐criterio se ha utilizado con el fin de evaluar las opciones de adaptación de acuerdo a las opiniones de las diferentes partes interesadas, el modelo de optimización ha tenido como fin analizar la combinación óptima de medidas de adaptación. El índice compuesto se ha utilizado para evaluar a nivel regional la implementación de medidas de adaptación en África. En general, los resultados del estudio ponen de relieve la gran importancia de considerar diferentes escalas espaciales a la hora de evaluar la implementación de medidas de adaptación al cambio climático. El comportamiento de los agricultores es diferente entre lugares considerados a una escala local relativamente pequeña, por lo que la generalización de los patrones del comportamiento a escalas regionales o globales resulta relativamente compleja. Los resultados obtenidos han permitido identificar factores determinantes tanto socioeconómicos como psicológicos y calcular su efecto sobre la adopción de medidas de adaptación. Además han proporcionado una mejor comprensión del distinto papel que desempeñan los cinco tipos de capital (natural, físico, financiero, social y humano) en la implementación de estrategias de adaptación. Con este trabajo se proporciona información de gran interés en los procesos de desarrollo de políticas destinadas a mejorar el apoyo de la sociedad a tomar medidas contra el cambio climático. Por último, en el análisis a nivel regional se desarrolla un índice compuesto que muestra la probabilidad de adoptar medidas de adaptación en las regiones de África y se analizan las causas que determinan dicha probabilidad de adopción de medidas. ABSTRACT Agriculture is and will continue to be one of the sectors most affected by climate change. Despite having demonstrated throughout history a great ability to adapt, agriculture today faces new challenges such as meeting growing food demands, developing sustainable agriculture and reducing greenhouse gas emissions. Adaptation policies planned on global, regional or local scales are ultimately implemented in decision‐making processes at the farm or individual level so adaptation potentials have to be set within the context of individual behaviour and regional institutions. Policy instruments can play a formative role in the adoption of such policies by addressing incentives/disincentives that influence farmer’s behaviour. Hence understanding farm‐level decision‐making processes and the influence of determinants of adoption is crucial when designing policies aimed at fostering adoption. This thesis seeks to analyse the factors that influence decision‐making by farmers in relation to the uptake of adaptation options. This work reviews the current knowledge and develops a methodological framework at local and regional level. Whilst the case studies at the local level are conducted with the purpose of exploring farmer’s behaviour towards adaptation the case study at the regional level attempts to up‐scale and generalise theory on adoption of farmlevel adaptation options. The two case studies at the local level (Doñana, Spain and Makueni, Kenya) encompass areas with different; climates, impacts of climate change, adaptation constraints and limits, levels of development, institutional support for agriculture and influence from public and private institutions. Whilst the Doñana Case Study represents an area plagued with water‐usage issues, set to be aggravated further by climate change, Makueni Case study exemplifies an area decidedly threatened by climate change where a lack of infrastructure and technology plays a crucial role in the uptake of adaptation options. The proposed framework is based on a wide range of approaches for collecting and analysing data. The approaches used for data collection include the implementation of surveys, interviews, stakeholder workshops, focus group discussions, a review of previous case studies, and public databases. The analytical methods include statistical approaches, multi criteria analysis for decision‐making, land use optimisation models, and a composite index based on public databases. Statistical approaches are used to assess the influence of socio‐economic and psychological factors on the adoption or support for adaptation measures. The statistical approaches used are logistic regressions, principal component analysis and structural equation modelling. Whilst a multi criteria analysis approach is used to evaluate adaptation options according to the different perspectives of stakeholders, the optimisation model analyses the optimal combination of adaptation options. The composite index is developed to assess adoption of adaptation measures in Africa. Overall, the results of the study highlight the importance of considering various scales when assessing adoption of adaptation measures to climate change. As farmer’s behaviour varies at a local scale there is elevated complexity when generalising behavioural patterns for farmers at regional or global scales. The results identify and estimate the effect of most relevant socioeconomic and psychological factors that influence adoption of adaptation measures to climate change. They also provide a better understanding of the role of the five types of capital (natural, physical, financial, social, and human) on the uptake of farm‐level adaptation options. These assessments of determinants help to explain adoption of climate change measures and provide helpful information in order to design polices aimed at enhancing societal support for adaptation policies. Finally the analysis at the regional level develops a composite index which suggests the likelihood of the regions in Africa to adopt farm‐level adaptation measures and analyses the main causes of this likelihood of adoption.

Housing plans and urban rehabilitation in Spain, 1992-2012

This paper explores the urban rehabilitation projects promoted by the Spanish Government between 1992 and 2012 through housing plans. The analysis is based on the comparison of programmes and estimations gathered in these plans with actual housing production within this period in order to find the connection between sectoral housing planning and real estate cycles in these last twenty years. During the period under review, six state housing plans, that were mainly focused on the promotion of newly-constructed state-subsidised housing, were developed, including the Areas of Integrated Rehabilitation programmes (ARI programmes). In spite of the relevance and growing complexity of these programmes, these played a subsidiary role in the government housing policy and were insignificant regarding the whole real estate production in this period.

CO2and Rn degassing from the natural analog of Campo de Calatrava(Spain): Implications for monitoring of CO2storage sites

Natural analogs offer a valuable opportunity to investigate the long-term impacts associated with thepotential leakage in geological storage of CO2.Degassing of CO2and radon isotopes (222Rn?220Rn) from soil, gas vents and thermal water dischargeswas investigated in the natural analog of Campo de Calatrava Volcanic Field (CCVF; Central Spain) todetermine the CO2?Rn relationships and to assess the role of CO2as carrier gas for radon. Furthermore,radon measurements to discriminate between shallow and deep gas sources were evaluated under theperspective of their applicability in monitoring programs of carbon storage projects.CO2flux as high as 5000 g m?2d?1and222Rn activities up to 430 kBq m?3were measured;220Rn activi-ties were one order of magnitude lower than those of222Rn. The222Rn/220Rn ratios were used to constrainthe source of the Campo de Calatrava soil gases since a positive correlation between radon isotopic ratiosand CO2fluxes was observed. Thus, in agreement with previous studies, our results indicate a deepmantle-related origin of CO2for both free and soil gases, suggesting that carbon dioxide is an efficientcarrier for Rn. Furthermore, it was ascertained that the increase of222Rn in the soil gases was likely pro-duced by two main processes: (i) direct transport by a carrier gas, i.e., CO2and (ii) generation at shallowlevel due to the presence of relatively high concentrations of dissolved U and Ra in the thermal aquiferof Campo de Calatrava.The diffuse CO2soil flux and radon isotopic surveys carried out in the Campo de Calatrava VolcanicFields can also be applicable to geochemical monitoring programs in CCS (Carbon Capture and Storage)areas as these parameters are useful to: (i) constrain CO2leakages once detected and (ii) monitor both theevolution of the leakages and the effectiveness of subsequent remediation activities. These measurementscan also conveniently be used to detect diffuse leakages.

Evaluation of the competitiveness of agri-food sector in the región of the Alcarria Conquense(Spain)

The agrifood industry, like other sectors, faces the ongoing challenge of improving their competitiveness in order to strengthen its market presence and cater to the growing global population. This research measures the competitiveness of the agrifood industry in the region of La Alcarria Conquense (Spain), in the framework of the evaluation of programs in the territory that have aimed at improving and enhancing this sector. Through building the competitiveness profiles (Porter, 1990) and cluster analysis we have identified six competitive strategy patterns in food companies in the region. In addition, we have analyzed each of the areas of competitiveness and we can identify the strengths and weaknesses of the sector, and identify recommendations for increasing the responsiveness of the territory. Among the defining characteristics are the lack of association, the limitation on payment systems or virtual absence of training and innovation. However, programs to support the sector are highly valued and reverse in the long-term viability of these companies.

Una arquitectura en el territorio. Naturaleza de los tipos de la edificación vernácula española: la casa tradicional soriana

La presente Tesis Doctoral trata de ofrecer el primer estudio monográfico global que se dedica a cierta arquitectura vernácula, de un territorio situado en el centro–norte de la Península Ibérica. Asentada a horcajadas entre la Meseta septentrional castellana y la depresión mediterránea del río Ebro, y repartida en comarcas vinculadas a la España del norte y áreas de aspecto más meridional, la provincia de Soria presenta una notable superposición de tradiciones culturales, que se reúnen para generar desarrollos edificatorios diversos. En relación con esa variedad esencial, algunos datos contenidos en este trabajo, y muchos resultados de sus análisis, podrían extrapolarse, sin excesivos forzamientos, a fenómenos constructivos paralelos, correspondientes a amplias regiones del país. Tras enunciar brevemente las razones y objetivos de la investigación, junto con sus planteamientos de partida y determinados principios metodológicos, se han presentado unos elementos preliminares: apartados en los que se describe el medio físico y el marco humano, dentro de los cuales la arquitectura tradicional se desarrolla. Seguidamente, da comienzo la monografía referida a una realidad construida compleja, cuya explicación se ha dividido, por causas instrumentales, en dos partes diferenciadas. En el primer desarrollo, de género esencialmente analítico, se presentan los objetos edificados desde la perspectiva de la realidad material y constructiva de sus partes componentes; o desde el punto de vista de las técnicas que, en su día, sirvieron para definir elementos edificatorios, y sistemas generales de ellos. En la sección segunda, de naturaleza algo más sintética, se muestran otros aspectos de las arquitecturas, tomadas ya globalmente, o asociadas en grandes agrupaciones y conjuntos. Describiendo, de modo conciso, los territorios y ambientes urbanos tradicionales, en los que unos u otros tipos comparecen. Y pasando después a exponer, ya in extenso, los rasgos comunes o más específicos de estos y aquellos modelos arquitectónicos, así como sus recíprocas vinculaciones o procesos tipológicos corrientes. A modo de discusión final, se incluyen determinadas conclusiones, deducidas de cuanto se ha expuesto a lo largo del texto. Tales resultados, de índole algo más teórica, nos ayudan a comprender cuál es la efectiva naturaleza de los tipos individuados en las construcciones populares sorianas, particularmente en aquéllas de uso doméstico. Carácter compartido, sin duda, con otros modelos que, para territorios de mayor dimensión, podrían llegar a describirse. Así, el fenómeno constructivo referenciado se contempla, más allá de su concreto alcance local, como una muestra representativa de realidades más vastas, de orden real y también conceptual, que se regirían principios análogos. ABSTRACT The present Dissertation brings the first monographic study dedicated to certain vernacular architecture located in the territory of Soria, at the Center-North of the Iberian Peninsula. Seated between the Northern Castilian Plateau and the Ebro River Mediterranean Depression, and spread into regions linked to Northern Spain and areas of more southern characteristics, the province of Soria presents a remarkable overlap of cultural traditions providing several building developments. That essential variety is widely shown throughout this work, where the information recovered and the analysis’ reports allow a feasable extrapolation that explain the constructive phenomena all over broad regions of the Spanish country. After stating briefly the aim of the research and its background, along with its methodological process, this work starts its task introducing some preliminary elements: the physical environment and the human context where traditional architecture takes place. Afterwards the text focuses on the different vernacular constructions in all their complexity, which exposure has been divided in two parts: the first one, analytic, shows the built objects from the point of view of the materials and each constructive element; or, in another way, the constructive techniques that define building elements in the overall constructive systems. The second section, more synthetic, shows other aspects of this kind of architecture considering all cases, attempting to a theoretical classification and describing the territory and the urban environments where these architectures are placed. As follows, common and specific features of the architectural types studied are widely shown, as well as reciprocal connections, or typological processes underlying. The final discussion extracts conclusions that help us to understand the effective nature of the types found in Soria’s vernacular construction, particularly those for domestic use. There is no doubt that these buildings share a common character with another ones belonging to different territories of larger dimensions, that could be concisely studied in other place. Thus, these studied constructive phenomena can be considered, beyond the specific local scope, as a sample for more complex architectures, both in conceptual and physical terms, which seem to be guided by similar principles.

Building resilience to water scarcity in Southern Spain: A case study of rice farming in Doñana protected wetlands

Agricultural water management needs to evolve in view of increased water scarcity, especially when farming and natural protected areas are closely linked. In the study site of Don?ana (southern Spain), water is shared by rice producers and a world heritage biodiversity ecosystem. Our aim is to contribute to defining adaptation strategies that may build resilience to increasing water scarcity and minimize water conflicts among agricultural and natural systems. The analytical framework links a participatory process with quantitative methods to prioritize the adaptation options. Bottom-up proposed adaptation measures are evaluated by a multi-criteria analysis (MCA) that includes both socioeconomic criteria and criteria of the ecosystem services affected by the adaptation options. Criteria weights are estimated by three different methods?analytic hierarchy process, Likert scale and equal weights?that are then compared. Finally, scores from an MCA are input into an optimization model used to determine the optimal land-use distribution in order to maximize utility and land-use diversification according to different scenarios of funds and water availability. While our results show a spectrum of perceptions of priorities among stakeholders, there is one overriding theme that is to define a way to restore part of the rice fields to natural wetlands. These results hold true under the current climate scenario and evenmore so under an increased water scarcity scenario.

Analysis and Potential of Use of Biomass Energy in Canary Islands, Spain

Biomass has always been associated with the development of the population in the Canary Islands as the first source of elemental energy that was in the archipelago and the main cause of deforestation of forests, which over the years has been replaced by forest fossil fuels. The Canary Islands store a large amount of energy in the form of biomass. This may be important on a small scale for the design of small power plants with similar fuels from agricultural activities, and these plants could supply rural areas that could have self-sufficiency energy. The problem with the Canary Islands for a boost in this achievement is to ensure the supply to the consumer centers or power plants for greater efficiency that must operate continuously, allowing them to have a resource with regularity, quality and at an acceptable cost. In the Canary Islands converge also a unique topography with a very rugged terrain that makes it greater difficult to use and significantly more expensive. In this work all these aspects are studied, giving conclusions, action paths and theoretical potentials.

Challenges for future of Natural Spaces of Canary Islands, Spain

The preservation of biodiversity is a fundamental objective of a ll policies related to a more sustainable development in any modern society. The rain forest and pine forests are two unique Canarian ecosystems with high importance to global biodiversity, holding a large number of endemic species and subspecies that is a priority to preserve. In this work the challenges that will face the natural areas of the Canary Islands are studied, as well as their fundamental value for economic and environmental development of the islands.

Air quality modeling and mortality impact of fine particles reduction policies in Spain

Background: In recent years, Spain has implemented a number of air quality control measures that are expected to lead to a future reduction in fine particle concentrations and an ensuing positive impact on public health. Objectives: We aimed to assess the impact on mortality attributable to a reduction in fine particle levels in Spain in 2014 in relation to the estimated level for 2007. Methods: To estimate exposure, we constructed fine particle distribution models for Spain for 2007 (reference scenario) and 2014 (projected scenario) with a spatial resolution of 16x16 km2. In a second step, we used the concentration-response functions proposed by cohort studies carried out in Europe (European Study of Cohorts for Air Pollution Effects and Rome longitudinal cohort) and North America (American Cancer Society cohort, Harvard Six Cities study and Canadian national cohort) to calculate the number of attributable annual deaths corresponding to all causes, all non-accidental causes, ischemic heart disease and lung cancer among persons aged over 25 years (2005-2007 mortality rate data). We examined the effect of the Spanish demographic shift in our analysis using 2007 and 2012 population figures. Results: Our model suggested that there would be a mean overall reduction in fine particle levels of 1mg/m3 by 2014. Taking into account 2007 population data, between 8 and 15 all-cause deaths per 100,000 population could be postponed annually by the expected reduction in fine particle levels. For specific subgroups, estimates varied from 10 to 30 deaths for all non-accidental causes, from 1 to 5 for lung cancer, and from 2 to 6 for ischemic heart disease. The expected burden of preventable mortality would be even higher in the future due to the Spanish population growth. Taking into account the population older than 30 years in 2012, the absolute mortality impact estimate would increase approximately by 18%. Conclusions: Effective implementation of air quality measures in Spain, in a scenario with a short-term projection, would amount to an appreciable decline infine particle concentrations, and this, in turn, would lead to notable health-related benefits. Recent European cohort studies strengthen the evidence of an association between long-term exposure to fine particles and health effects, and could enhance the health impact quantification in Europe. Air quality models can contribute to improved assessment of air pollution health impact estimates, particularly in study areas without air pollution monitoring data.

Cross Ventilation CFD Modeling: Characterization and Study of Different Façade Openings Configurations at the Refurbishment of a Residential Building in Málaga (Spain)

The opening of new windows on the façade is proposed as a refurbishment strategy in an existing building in Málaga to facilitate cross ventilation of dwellings. The building is a residential block of 140 public housing units for rent for people with low income in Málaga (Spain), property of the City Council. By modeling with Computational Fluid Dynamics (CFD), eleven configurations of openings are studied in two different areas of the main housing type of the building. The quantity of introduced/extracted air into/from the room and the generated airflow patterns are obtained. The modeling allows comparing the different openings configurations to determine the most appropriate ventilation option for every room.

Development and assessment of traffic-related emission abatement measures for the Madrid city (Spain) through the WRF-SMOKE-CMAQ modelling system

The achievement of the limit values established in the European legislation pose an important handicap for large urban areas with intense road traffic, such as Madrid (Spain). Despite permanent measures included in air quality plans it is important to assess additional measures that may be temporally applied under unfavourable conditions. This paper reports on the simulation of different traffic restriction strategies in Madrid for high-pollution episodes.

Analysis of Contributions to NO2 Ambient Air Quality Levels in Madrid City (Spain) through Modeling. Implications for the Development of Policies and Air Quality Monitoring

As environmental standards become more stringent (e.g. European Directive 2008/50/EC), more reliable and sophisticated modeling tools are needed to simulate measures and plans that may effectively tackle air quality exceedances, common in large cities across Europe, particularly for NO2. Modeling air quality in urban areas is rather complex since observed concentration values are a consequence of the interaction of multiple sources and processes that involve a wide range of spatial and temporal scales. Besides a consistent and robust multi-scale modeling system, comprehensive and flexible emission inventories are needed. This paper discusses the application of the WRF-SMOKE-CMAQ system to the Madrid city (Spain) to assess the contribution of the main emitting sectors in the region. A detailed emission inventory was compiled for this purpose. This inventory relies on bottom-up methods for the most important sources. It is coupled with the regional traffic model and it makes use of an extensive database of industrial, commercial and residential combustion plants. Less relevant sources are downscaled from national or regional inventories. This paper reports the methodology and main results of the source apportionment study performed to understand the origin of pollution (main sectors and geographical areas) and define clear targets for the abatement strategy. Finally the structure of the air quality monitoring is analyzed and discussed to identify options to improve the monitoring strategy not only in the Madrid city but the whole metropolitan area.

Native bradyrhizobial symbionts of Lupinus mariae-josephae, a unique endemism thriving in alkaline soils in Eastern Spain

Lupinus mariae-josephae (Lmj) es una especie de lupino endémica de una pequeña y específica área de Comunidad Valenciana (Este de España), donde prospera en suelos alcalinoscalcáreos, un hábitat singular para los altramuces, que crecen preferentemente en suelos ácidos o neutros. Esto hace de Lmj una especie de lupino única. Cuando se inició este trabajo, la extensión conocida de este endemismo abarcaba unos 700 kilómetros cuadrados, confinados en la provincia de Valencia. En esta área, Lmj prospera en pequeñas poblaciones aisladas que contienen un número reducido de plantas por lo que se la consideró una especie en peligro de extinción. Todos los esfuerzos, utilizando estrategias clásicas dirigidas a ampliar el área de crecimiento de Lmj y garantizar su conservación, han tenido un éxito limitado. El trabajo que se presenta está dirigido a mejorar el conocimiento de la ecología de Lmj, en particular la interacción simbiótica que establece con bacterias del suelo denominadas rizobios y se centra en la caracterización fenotípica, filogenética y genómica de esos rizobios. También se investiga la posible contribución de la simbiosis en mejorar la conservación de Lmj. Para este fin, se han estudiado diferentes aspectos que se describen a continuación. El primero objetivo se centró en aislar y estudiar de la diversidad genética de las bacterias endosimbióticas de Lmj. . Se realizó un análisis filogenético de genes esenciales que mostró que las cepas de Lmj pertenecen al género Bradyrhizobium y que presentan una gran diversidad con características fenotípicas y simbióticas diferentes de cepas de Bradyrhizobium que nodulan otras especies de lupinos nativos de España (cepas ISLU). Las cepas estudiadas se dividieron en dos grupos (Clado I y Clado II). El Clado I, incluye a las cepas Lmj, definiendo un nuevo linaje, filogenéticamente relacionado con otras especies de Bradyrhizobium, como B. jicamae y B. elkanii. El Clado II contiene cepas ISLU relacionadas con cepas de B. canariense y B. japonicum que establecen simbiosis con lupinos de suelos ácidos. Otro análisis filogenético basado en genes simbióticos, distribuyó las cepas de Lmj en sólo dos grupos diferentes. La singularidad y gran diversidad de estas cepas en una pequeña área geográfica, hacen de este, un atractivo sistema para el estudio de la evolución y adaptación de las bacterias simbióticas a su respectiva planta huésped. Adicionalmente, se estudio la presencia de bacterias capaces de nodular Lmj en suelos básicos de Chiapas, México. Sorprendentemente, estos suelos contienen bacterias capaces establecer interacciones simbióticas eficientes con Lmj en ensayos de invernadero. A continuación se investigó la taxonomía de los endosimbiontes de Lmj analizando la secuencia de cuatro genes esenciales (16S rRNA, recA, glnII y atpD) y el promedio de identidad de nucleótidos de genomas completos de algunas cepas representativas de la diversidad (ANIm). Se identificaron nuevas especies de Bradyrhizobium dentro del Clado I y se definió una de ellas: 'Bradyrhizobium valentinum' sp. nov (cepa tipo LmjM3T = CECT 8364T, LMG 2761T). También se abordó cómo conservar Lmj en su hábitat natural mediante inoculación con alguna de las cepas aisladas. Se demostró la ausencia de bacterias capaces de nodular Lmj en suelos rojos alcalinos o ‘‘terra rossa’’ de la Península Ibérica y Baleares. Dos cepas, altamente eficientes en cuanto a la fijación de nitrógeno, LmjC y LmjM3T, fueron seleccionadas para ser empleadas como inoculantes. Dos experimentos de campo llevados a cabo en años consecutivos en áreas con características edafoclimáticas similares a las que presentan las poblaciones de Lmj, lograron la reproducción exitosa de la planta. Se concluyó que un ciclo reproductivo exitoso de Lmj es absolutamente dependiente de la inoculación con sus simbiontes naturales y que la simbiosis debe ser considerada un factor esencial en estrategias de conservación de leguminosas en peligro. La obtención de varias secuencias genómicas de cepas aisladas de Lmj y de otras cepas de Bradyrhizobium reveló una alta similitud entre los genomas de las cepas del Clado I, y permitió la identificación de cinco posibles nuevas especies. Además, se estudiaron tres agrupaciones de genes relacionados con la simbiosis (nod, nif y fix) definiendo un nuevo linaje para las cepas de Lmj, diferente del symbiovar “genistearum” de B. canariense y B. japonicum. La baja diversidad encontrada en el análisis filogenético de los genes simbióticos contrasta con la gran diversidad asociada a genes esenciales. La presencia de plásmidos en cepas del género Bradyrhizobium ha sido descrita en muy pocas ocasiones, sin embargo el análisis de la secuencia genómica de la cepa ISLU101, aislada de Lupinus angustifolius, reveló la presencia de un origen de replicación extracromosómico homólogo al operón repABC, presente en el plásmido de Bradyrhizobium sp BTAi1. Gracias a esta secuencia se identificaron genes homólogos en 19 de 72 cepas ISLU. Filogenéticamente, las secuencias de repABC se agruparon en un grupo monofilético con las de pBTAi1 y separadas de los rizobios de crecimiento rápido. Finalmente, se identificaron sistemas de secreción de proteínas de tipo III (T3SS) en nueve genomas de cepas de Lmj. Los T3SS pueden inyectar proteínas efectoras al interior de células vegetales. Su presencia en rizobios se ha relacionado con la gama de hospedador que pueden nodular y puede tener un efecto beneficioso, neutro o perjudicial en la simbiosis. Los T3SS de las cepas de Lmj codifican para una proteína efectora similar a NopE, un efector dependiente de T3SS descrito en B. diazoefficiens USDA 110T. La proteína NopE de la cepa LmjC se ha caracterizado bioquímicamente. ABSTRACT Lupinus mariae-josephae (Lmj) is a lupine species endemic of a unique small area in Valencia region (Eastern Spain) where the lupine plants thrive in alkaline-limed soils, which preferentially grow in acid or neutral soils. This is the type of soils native lupines of Spain. When this work was initiated, the extension of the endemic area of Lmj was of about 700 squared kilometers confined to the Valencia province. In this area, Lmj thrives in small, isolated patches containing a reduced number of plants, and points to an endemism that can easily became endangered or extinct. Consequently, the Valencia Community authorities gave a ‘‘microreserve” status for conservation of the species. All efforts, using classical strategies directed to extend the area of Lmj growth and ensure its conservation have been so far unsuccessful. The work presented here is directed to improve our knowledge of Lmj ecology and it is centered in the characterization of the rhizobial symbiosis by phenotypic, phylogenetic and genomic analysis as well as in investigate the potential contribution of the symbiosis to improve its conservation. To this end, five different topics have been studied, and results are briefly described here. Extensive details can be followed en the attached, published articles. The first topic deals with the indigenous rhizobial symbionts of the Lmj endemism, and its genetic diversity was investigated. The Lmj root symbionts belong to the Bradyrhizobium genus, and phylogenetic analysis based on core genes identified a large diversity of Bradyrhizobium strains with phenotypic and symbiotic characteristics different from rhizobia nodulating other Lupinus spp. native of Spain. The strains were split in two clades. Clade II contained strains close to classical B. canariense and B. japonicum lineages that establish symbioses with lupines in acid soils of the Mediterranean area. Clade I included Lmj strains that define a new lineage, close to other Bradyrhizobium species as B. jicamae and B. elkanii. The phylogenetic analysis based on symbiotic genes identified only two distinct clusters. The singularity and large diversity of these strains in such a small geographical area makes this an attractive system for studying the evolution and adaptation of the rhizobial symbiont to the plant host. Additionally, the presence of bacteria able to nodulate Lmj in basic soils from Chiapas, Mexico was investigated. Surprisingly, these soils contain bacteria able to effectively nodulate and fix nitrogen with Lmj plants in greenhouse assays. In the second topic, the taxonomic status of the endosymbiotic bacteria of Lmj from Valencia endemism and Chiapas was investigated. Results from phylogenetic analysis of core genes and Average Nucleotide Identity (ANIm) using draft genomic sequences identified new Bradyrhizobium species within strains of Clade I of Lmj endosymbiotic bacteria. Only one of these potentially new species has been defined, meanwhile the others are under process of characterization. The name ‘Bradyrhizobium valentinum’ sp. nov. was proposed for the defined species (type strain LmjM3T= CECT 8364T, LMG 2761T). The third topic was directed to conservation of endangered Lmj in its natural habitat. The relevant conclusion of this experimentation is that the symbiosis should be considered as a relevant factor in the conservation strategies for endangered legumes. First, we showed absence of bacteria able to nodulate Lmj in all the inspected ‘‘terra rossa’’ or alkaline red soils of the Iberian Peninsula and Balearic Islands. Then, two efficient nitrogen fixing strains with Lmj plants, LmjC and LmjM3T, were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural Lmj populations, and successful reproduction of the plant was achieved. The relevant conclusion from these assays was that the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia The forth topic deep into the analysis of the genomic of Lmj representative strains. To this end, draft genomic sequences of selected Lmj strains and type strains of Bradyrhizobium spp. were assembled. The comparison analysis of the draft genomic sequences of Lmj strains and related Bradyrhizobium species grouped in Clade I, revealed a high genomic homology among them, and allowed the definition of five potentially new species of Lmj nodulating bacteria. Also, based on the available draft genomic sequences, only three clusters of nod, fix and nif genes from Lmj strains were identified and showed to define a new symbiotic lineage, distant from that of B. canariense and B. japonicum bv. genistearum. The low diversity exhibited by the phylogenetic analysis of symbiotic genes contrast with the large diversity of strains as regards the housekeeping genes analyzed. Besides, the genomic analysis of a Lupinus angustifolius strain ISLU101, revealed the presence of an extrachromosomal replication origin homologous to repABC cluster from plasmid present in Bradyrhizobium spp BTAi1. This repABC cluster gene sequence allowed the identification of extrachromosomic replication origin in 19 out of 72 Bradyrhizobium strains from Lupinus spp., a highly significant result since the absence of plasmids in the Bradyrhizobium genus was traditionally assumed. The repABC gene sequences of these strains grouped them in a unique monophyletic group, related to B. sp. BTAi1 plasmid, but differentiated from the repABC gene cluster of plasmids in fast growing rhizobium strains. The last topic was focused on characterization of type III secreted effectors present in Lmj endosymbiotic bacteria. Type III secretion systems (T3SS) are specialized protein export machineries which can deliver effector proteins into plant cells. The presence of T3SS in rhizobia has frequently been related to the symbiotic nodulation host-range and may have a beneficial or detrimental effect on the symbiosis with legumes. In this context, the presence of T3SS in genomes of nine Lmj strains was investigated, and it was shown the presence of clusters encoding NopE type III-secreted protein similar to the NopE1 and NopE2 of B. diazoefficiens USDA 110T. The putative NopE protein of LmjC strain is at present being characterized regarding its structure and function.

Viability of the biochar production from different manure wastes in the Amblés Valley (Ávila, Spain)

In the last years, intensive animal husbandry production has led to a large concentration of animals in small areas. This has resulted in the production of excessive amounts of manures with insufficient nearby land for application. One of this areas is the Amblés Valley located in the centre of Spain, near to Ávila city, with an extension of 167472 ha of which 88.9% is agricultural land. This valley has an important livestock focused on pig, cattle, chicken production which is associated with the generation of more than 200,000 t/year of manure. There are a number of environmental problems associated with these intensive agricultural systems, including N and P pollution of water bodies, methane emissions and odour pollution. These serious environmental threats are called for innovative environmental management approaches. A feasible technology for the management of manures, offering a potential to valorise these wastes, is pyrolysis, which results in the production of biochar. The objective of this work is evaluated the technical and economic feasibility of the production of biochar in Amblés Valley (Spain).