La geoquímica de los sedimentos de llanuras de inundación como guía para la valoración ambiental en España

En la presente tesis se estudian los contenidos geoquímicos de los sedimentos de llanura de inundación en diversas cuencas fluviales seleccionadas con el objetivo de contribuir a un mejor conocimiento de sus condiciones medioambientales. En cada cuenca, se ha muestreado un perfil vertical de llanura de inundación, dividiéndolo en tramos que generalmente corresponden a diferentes episodios de inundación. Estos sedimentos se depositan durante los episodios de crecidas una vez que la corriente sobrepasa los límites del canal. Se caracterizan normalmente por tener un tamaño de grano fino y una estructura en capas horizontales que corresponden a los sucesivos episodios de inundación. Muestran dos ventajas principales con respecto a otros medios de muestreo en geoquímica como suelos o sedimentos de corriente: • Pueden almacenar sedimento antiguo así como actual, con lo que se puede estudiar la historia geoquímica de una zona específica en un mismo punto de muestreo (perfil vertical). • Los sedimentos de llanuras de inundación son capaces de caracterizar grandes áreas de drenaje. El origen de los sedimentos es más diverso que en el sedimento de corriente, debido a las mayores áreas de donde proceden las aguas de las avenidas. Las cuencas han sido seleccionadas según las actividades antropogénicas que en ellas se llevan a cabo, en concreto, actividades urbanas e industriales, minería y agricultura. Así mismo, se han estudiado, como referencia, dos cuencas donde no se espera encontrar ningún tipo de actividad contaminante. Una vez hecha la selección, los sedimentos aluviales de las cuencas se han estudiado cuidadosamente para asegurar que no existen depósitos de acreción lateral en el punto seleccionado. Posteriormente se ha procedido al muestreo del perfil vertical. Las muestras han sido analizadas mediante ICP-MS (ataque total) e INAA para conocer los contenidos totales de los elementos traza y mayoritarios. Los análisis de la fracción extraíble se han llevado a cabo mediante ICP-MS (ataque con agua regia). Así mismo, algunas muestras seleccionadas han sido sometidas a una extracción secuencial para un estudio más detallado. La presencia de materia orgánica ha sido estimada mediante el análisis de Carbono Orgánico Total (TOC). Finalmente, se ha llevado a cabo un análisis de isótopos de Pb en muestras escogidas en los perfiles, con el objetivo de hacer una evaluación ambiental. Los contenidos metálicos aumentan hacia la superficie en algunos de los perfiles, mientras en otros muestran una distribución muy constante exceptuando algún nivel específico con un aumento de los contenidos de la mayoría de los metales. Ha sido posible determinar la influencia de las actividades antropogénicas en algunos de los perfiles. Aquellos que pertenecen a cuencas mineras, urbanas o industrializadas muestran generalmente altos contenidos en elementos metálicos. Es el caso de los perfiles muestreados en los ríos Odiel y Tinto, Besaya, Besós y Manzanares. Algunos de estos perfiles pueden incluso correlacionarse con periodos de tiempo en los que ha tenido lugar una actividad antropogénica más intensa. Los perfiles que mejor se correlacionan con la actividad antropogénica de la cuenca son el perfil de Rivas en el río Manzanares (Madrid), que refleja un crecimiento de la contaminación producida por las actividades urbana e industrial en las últimas décadas, y el río Tinto, que muestra un crecimiento llamativo de los contenidos en su mayoría metálicos que puede estar relacionado con el incremento de la actividad minera que tuvo lugar hace aproximadamente 125 años. El análisis de los isótopos de Pb ha resultado ser una herramienta útil en la evaluación ambiental de estos sedimentos. Con este estudio y mediante la comparación con fuentes naturales y antropogénicas, ha sido posible diferenciar las muestras afectadas por diferentes fuentes de plomo, así como detectar las más afectadas antropogénicamente. ABSTRACT The geochemical composition of overbank sediments of some selected river basins is studied in this thesis in order to contribute to a better knowledge of the environmental conditions surrounding them. In each basin a vertical overbank profile has been sampled, dividing it into stretches that usually correspond to different flood events. The overbank sediments are those deposited during a flood event once the flow spills over the channel banks. They are usually characterized by a very fine grain size and a structure of horizontal layers, which correspond to successive flood events. These sediments show two main advantages regarding other sampling media in geochemistry, like soils or stream sediments: • They can store sediment deposited in the past as well as in current times, so that the history of a specific location can be studied at the very same point (vertical profile). • The overbank sediments are able to characterize a large drainage area. The origin of the sediment is wider than in the stream sediments due to the larger areas where the flood water comes from. The basins have been selected depending on the anthropogenic activities developed in them, namely, urban and industrial activities, mining activities and agricultural activities. As well, two pristine basins have been studied as a reference. Afterwards, the alluvial sediments in the basins have been carefully studied in order to sample a vertical profile and make sure that lateral accretion materials are not present in the profile. The samples have been analysed by ICP-MS (total digestion) and INAA to know the total contents of trace and major elements. Analysis of the mobile fraction has been carried out by ICP-MS (aqua regia); as well some of the samples have been subjected to sequential extraction for a more detailed study. The presence of organic matter has been estimated by the analysis of the Total Organic Carbon (TOC). Finally, a lead isotope analysis of some of the samples in the profiles was carried out in order to make an environmental assessment. Metal contents grow towards the surface in some of the profiles, while others show a very steady distribution, except for some of them with a growth of most of the metals in a specific level. XI It has been possible to determine the influence of the anthropogenic activities in some of the profiles. The ones that belong to mining and urban or industrialized basins show generally high contents of metal elements. This is the case of the profiles sampled in the Odiel and Tinto Rivers, the Besaya River, the Besós River and the Manzanares River. Some of these profiles can even correlate with the periods of time when a more intense activity in their respective basins has taken place. The profiles which best correlate with the anthropogenic activity are the Rivas profile in the Manzanares River, which reflects a growth of the pollution produced by urban and industrial activities in the city of Madrid in the last decades and the Tinto profile, which shows a very dramatic growth of the elemental contents (mostly metals) which can be related to the increase of the mining activities that took place in the last 125 years. The analysis of lead isotopes has turned out to be a powerful tool in the environmental assessment in this type of sediments. With this study and through the comparison with natural and anthropogenic sources, it has been possible to determine samples affected by different sources of lead and to detect the most anthropogenicaly affected ones.

Geochemical fingerprints and controls in the sediments of an urban river: River Manzanares, Madrid (Spain)

The geochemical fingerprint of sediment retrieved from the banks of the River Manzanares as it passes through the City of Madrid is presented here. The river collects the effluent water from several Waste Water Treatment (WWT) plants in and around the city, such that, at low flows, up to 60% of the flow has been treated. A total of 18 bank-sediment cores were collected along the course of the river, down to its confluence with the Jarama river, to the south–east of Madrid. Trace and major elements in each sample were extracted following a double protocol: (a) “Total” digestion with HNO3, HClO4 and HF; (b) “Weak” digestion with sodium acetate buffered to pH=5 with acetic acid, under constant stirring. The digests thus obtained were subsequently analysed by ICP-AES, except for Hg which was extracted with aqua regia and sodium chloride-hydroxylamine sulfate, and analysed by Cold Vapour-AAS. X-ray diffraction was additionally employed to determine the mineralogical composition of the samples. Uni- and multivariate analyses of the chemical data reveal the influence of Madrid on the geochemistry of Manzanares' sediments, clearly manifested by a marked increase in the concentration of typically “urban” elements Ag, Cr, Cu, Pb and Zn, downstream of the intersection of the river with the city's perimeter. The highest concentrations of these elements appear to be associated with illegal or accidental dumping of waste materials, and with the uncontrolled incorporation of untreated urban runoff to the river. The natural matrix of the sediment is characterised by fairly constant concentrations of Ce, La and Y, whereas changes in the lithology intersected by the river cause corresponding variations in Ca–Mg and Al–Na contents. In the final stretch of the river, the presence of carbonate materials seems to exert a strong geochemical control on the amount of Zn and, to a lesser extent, Cu immobilised in the sediments. This fact suggests that a variable but significant proportion of both elements may be susceptible to reincorporation in the aqueous phase under realistic environmental conditions.

Spatial variability patterns of some Vertisol properties at a field scale using standardized data

Spatial variability of Vertisol properties is relevant for identifying those zones with physical degradation. In this sense, one has to face the problem of identifying the origin and distribution of spatial variability patterns. The objectives of the present work were (i) to quantify the spatial structure of different physical properties collected from a Vertisol, (ii) to search for potential correlations between different spatial patterns and (iii) to identify relevant components through multivariate spatial analysis. The study was conducted on a Vertisol (Typic Hapludert) dedicated to sugarcane (Saccharum officinarum L.) production during the last sixty years. We used six soil properties collected from a squared grid (225 points) (penetrometer resistance (PR), total porosity, fragmentation dimension (Df), vertical electrical conductivity (ECv), horizontal electrical conductivity (ECh) and soil water content (WC)). All the original data sets were z-transformed before geostatistical analysis. Three different types of semivariogram models were necessary for fitting individual experimental semivariograms. This suggests the different natures of spatial variability patterns. Soil water content rendered the largest nugget effect (C0 = 0.933) while soil total porosity showed the largest range of spatial correlation (A = 43.92 m). The bivariate geostatistical analysis also rendered significant cross-semivariance between different paired soil properties. However, four different semivariogram models were required in that case. This indicates an underlying co-regionalization between different soil properties, which is of interest for delineating management zones within sugarcane fields. Cross-semivariograms showed larger correlation ranges than individual, univariate, semivariograms (A ≥ 29 m). All the findings were supported by multivariate spatial analysis, which showed the influence of soil tillage operations, harvesting machinery and irrigation water distribution on the status of the investigated area.

The relationship between soil geochemistry and the bioaccessibility of trace elements in playground soil

A total of 32 samples of surficial soil were collected from 16 playground areas in Madrid (Spain), in order to investigate the importance of the geochemistry of the soil on subsequent bioaccessibility of trace elements. The in vitro bioaccessibility of As, Co, Cr, Cu, Ni, Pb and Zn was evaluated by means of two extraction processes that simulate the gastric environment and one that reproduces a gastric + intestinal digestion sequence. The results of the in vitro bioaccessibility were compared against aqua regia extractions (“total” concentration), and it was found that total concentrations of As, Cu, Pb and Zn were double those of bioaccessible values, whilst that of Cr was ten times higher. Whereas the results of the gastric + intestinal extraction were affected by a high uncertainty, both gastric methods offered very similar and consistent results, with bioaccessibilities following the order: As = Cu = Pb = Zn > Co > Ni > Cr, and ranging from 63 to 7 %. Selected soil properties including pH, organic matter, Fe and CaCO3 content were determined to assess their influence on trace element bioaccessibility, and it was found that Cu, Pb and Zn were predominantly bound to organic matter and, to a lesser extent, Fe oxides. The former fraction was readily accessible in the gastric solution, whereas Fe oxides seemed to recapture negatively charged chloride complexes of these elements in the gastric solution, lowering their bioaccessibility. The homogeneous pH of the playground soils included in the study does not influence trace element bioaccessibility to any significant extent except for Cr, where the very low gastric accessibility seems to be related to the strongly pH-dependent formation of complexes with organic matter. The results for As, which have been previously described and discussed in detail in Mingot et al. (Chemosphere 84: 1386–1391, 2011), indicate a high gastric bioaccessibility for this element as a consequence of its strong association with calcium carbonate and the ease with which these bonds are broken in the gastric solution. The calculation of risk assessments are therefore dependant on the methodology used and the specific environment they address. This has impacts on management strategies formulated to ensure that the most vulnerable of society, children, can live and play without adverse consequences to their health.

Assessment of oral bioaccessibility of arsenic in playground soil in Madrid (Spain): A three-method comparison and implications for risk assessment

Three methodologies to assess As bioaccessibility were evaluated using playgroundsoil collected from 16 playgrounds in Madrid, Spain: two (Simplified Bioaccessibility Extraction Test: SBET, and hydrochloric acid-extraction: HCl) assess gastric-only bioaccessibility and the third (Physiologically Based Extraction Test: PBET) evaluates mouth–gastric–intestinal bioaccessibility. Aqua regia-extractable (pseudo total) As contents, which are routinely employed in riskassessments, were used as the reference to establish the following percentages of bioaccessibility: SBET – 63.1; HCl – 51.8; PBET – 41.6, the highest values associated with the gastric-only extractions. For Madridplaygroundsoils – characterised by a very uniform, weakly alkaline pH, and low Fe oxide and organic matter contents – the statistical analysis of the results indicates that, in contrast with other studies, the highest percentage of As in the samples was bound to carbonates and/or present as calcium arsenate. As opposed to the As bound to Fe oxides, this As is readily released in the gastric environment as the carbonate matrix is decomposed and calcium arsenate is dissolved, but some of it is subsequently sequestered in unavailable forms as the pH is raised to 5.5 to mimic intestinal conditions. The HCl extraction can be used as a simple and reliable (i.e. low residual standard error) proxy for the more expensive, time consuming, and error-prone PBET methodology. The HCl method would essentially halve the estimate of carcinogenic risk for children playing in Madridplaygroundsoils, providing a more representative value of associated risk than the pseudo-total concentrations used at present

Environmental assay on the effect of poultry manure application on soil organisms in agroecosystems

This paper reports the effects produced on the organisms of the soil (plants, invertebrates and microorganisms), after the application of two types of poultry manure (sawdust and straw bed) on an agricultural land. The test was made using a terrestrial microcosm, Multi-Species Soil System (MS3) developed in INIA. There was no difference in the germination for any of the three species of plants considered in the study. The biomass was increased in the wheat (Triticum aestivum) coming from ground treated with both kinds of poultry manure. Oilseed rape (Brasica rapa) was not affected and regarding vetch (Vicia sativa) only straw poultry manure showed significant difference. For length only Vicia sativa was affected showing a reduction when straw was exposed to poultry manure. When the effect on invertebrates was studied, we observed a reduction in the number of worms during the test, especially from the ground control (13.7%), higher than in the ground with sawdust poultry manure (6.7%), whereas in the ground with straw poultry manure, there was no reduction. The biomass was affected and at the end of the test it was observed that while the reduction of worms in the ground control was about 48%, the number of those that were in the ground with sawdust poultry manure or straw poultry manure decreased by 41% and 22% respectively. Finally, the effects on microorganisms showed that the enzymatic activities: dehydrogenase (DH) and phosphatase and basal respiration rate increased at the beginning of the test, and the differences were statistically significant compared with the values of the control group. During the test, all these parameters decreased (except DH activities) but they were always higher than in the ground control. This is why it is possible to deduce that the contribution of poultry manure caused an improvement in the conditions of fertilization and also for the soil.

Fractal scaling of apparent soil moisture estimated from vertical planes of Vertisol pit images

Image analysis could be a useful tool for investigating the spatial patterns of apparent soil moisture at multiple resolutions. The objectives of the present work were (i) to define apparent soil moisture patterns from vertical planes of Vertisol pit images and (ii) to describe the scaling of apparent soil moisture distribution using fractal parameters. Twelve soil pits (0.70 m long × 0.60 m width × 0.30 m depth) were excavated on a bare Mazic Pellic Vertisol. Six of them were excavated in April/2011 and six pits were established in May/2011 after 3 days of a moderate rainfall event. Digital photographs were taken from each Vertisol pit using a Kodak™ digital camera. The mean image size was 1600 × 945 pixels with one physical pixel ≈373 μm of the photographed soil pit. Each soil image was analyzed using two fractal scaling exponents, box counting (capacity) dimension (DBC) and interface fractal dimension (Di), and three prefractal scaling coefficients, the total number of boxes intercepting the foreground pattern at a unit scale (A), fractal lacunarity at the unit scale (Λ1) and Shannon entropy at the unit scale (S1). All the scaling parameters identified significant differences between both sets of spatial patterns. Fractal lacunarity was the best discriminator between apparent soil moisture patterns. Soil image interpretation with fractal exponents and prefractal coefficients can be incorporated within a site-specific agriculture toolbox. While fractal exponents convey information on space filling characteristics of the pattern, prefractal coefficients represent the investigated soil property as seen through a higher resolution microscope. In spite of some computational and practical limitations, image analysis of apparent soil moisture patterns could be used in connection with traditional soil moisture sampling, which always renders punctual estimates

Seasonal evolution of soil respiration in a mixed forestry plantation of walnuts (Juglans × intermedia Carr.) and alders (Alnus cordata (Loisel.) Duby) in Domaine de Restinclières (France)

The ecological intensification of crops is proposed as a solution to the growing demand of agricultural and forest resources, in opposition to intensive monocultures. The introduction of mixed cultures as mixtures between nitrogen fixing species and non nitrogen fixing species intended to increase crop yield as a result of an improvement of the available nitrogen and phosphorus in soil. Relationship between crops have received little attention despite the wide range of advantages that confers species diversity to these systems, such as increased productivity, resilience to disruption and ecological sustainability. Forests and forestry plantations can develop an important role in storing carbon in their tissues, especially in wood which become into durable product. A simplifying parameter to analyze the amount allocated carbon by plantation is the TBCA (total belowground carbon allocation), whereby, for short periods and mature plantations, is admitted as the subtraction between soil carbon efflux and litterfall. Soil respiration depends on a wide range of factors, such as soil temperature and soil water content, soil fertility, presence and type of vegetation, among others. The studied orchard is a mixed forestry plantation of hybrid walnuts(Juglans × intermedia Carr.) for wood and alders (Alnus cordata (Loisel.) Duby.), a nitrogen fixing specie through the actinomycete Frankia alni ((Woronin, 1866) Von Tubeuf 1895). The study area is sited at Restinclières, a green area near Montpellier (South of France). In the present work, soil respiration varied greatly throughout the year, mainly influenced by soil temperature. Soil water content did not significantly influence the response of soil respiration as it was constant during the measurement period and under no water stress conditions. Distance between nearest walnut and measurement was also a highly influential factor in soil respiration. Generally there was a decreasing trend in soil respiration when the distance to the nearest tree increased. It was also analyzed the response of soil respiration according to alder presence and fertilizer management (50 kg N·ha-1·año-1 from 1999 to 2010). None of these treatments significantly influenced soil respiration, although previous studies noticed an inhibition in rates of soil respiration under fertilized conditions and high rates of available nitrogen. However, treatments without fertilization and without alder presence obtained higher respiration rates in those cases with significant differences. The lack of significant differences between treatments may be due to the high coefficient of variation experienced by soil respiration measurements. Finally an asynchronous fluctuation was observed between soil respiration and litterfall during senescence period. This is possibly due to the slowdown in the emission of exudates by roots during senescence period, which are largely related to microbial activity.

Fractal scaling of apparent soil moisture estimated from vertical planesof Vertisol pit images

Image analysis could be a useful tool for investigating the spatial patterns of apparent soil moisture at multiple resolutions. The objectives of the present work were (i) to define apparent soil moisture patterns from vertical planes of Vertisol pit images and (ii) to describe the scaling of apparent soil moisture distribution using fractal parameters. Twelve soil pits (0.70 m long × 0.60 m width × 0.30 m depth) were excavated on a bare Mazic Pellic Vertisol. Six of them were excavated in April/2011 and six pits were established in May/2011 after 3 days of a moderate rainfall event. Digital photographs were taken from each Vertisol pit using a Kodak? digital camera. The mean image size was 1600 × 945 pixels with one physical pixel ?373 ?m of the photographed soil pit. Each soil image was analyzed using two fractal scaling exponents, box counting (capacity) dimension (DBC) and interface fractal dimension (Di), and three prefractal scaling coefficients, the total number of boxes intercepting the foreground pattern at a unit scale (A), fractal lacunarity at the unit scale (?1) and Shannon entropy at the unit scale (S1). All the scaling parameters identified significant differences between both sets of spatial patterns. Fractal lacunarity was the best discriminator between apparent soil moisture patterns. Soil image interpretation with fractal exponents and prefractal coefficients can be incorporated within a site-specific agriculture toolbox. While fractal exponents convey information on space filling characteristics of the pattern, prefractal coefficients represent the investigated soil property as seen through a higher resolution microscope. In spite of some computational and practical limitations, image analysis of apparent soil moisture patterns could be used in connection with traditional soil moisture sampling, which always renders punctual estimates.

The role of cover crops in irrigated systems: water balance, nitrate leaching and soil mineral nitrogen accumulation

Soil salinity and salt leaching are a risk for sustainable agricultural production in many irrigated areas. This study was conducted over 3.5 years to determine how replacing the usual winter fallow with a cover crop (CC) affects soil salt accumulation and salt leaching in irrigated systems. Treatments studied during the period between summer crops were: barley (Hordeum vulgare L.), vetch (Vicia villosa L.) and fallow. Soil water content was monitored daily to a depth of 1.3 m and used with the numerical model WAVE to calculate drainage. Electrical conductivity (EC) was measured in soil solutions periodically, and in the soil saturated paste extracts before sowing CC and maize. Salt leaching was calculated multiplying drainage by total dissolved salts in the soil solution, and use to obtain a salt balance. Total salt leaching over the four winter fallow periods was 26 Mg ha−1, whereas less than 18 Mg ha−1 in the presence of a CC. Periods of salt gain occurred more often in the CC than in the fallow. By the end of the experiment, net salt losses occurred in all treatments, owing to occasional periods of heavy rainfall. The CC were more prone than the fallow to reduce soil salt accumulation during the early growth stages of the subsequent cash crop.

The role of cover crops in irrigated systems: Soil salinity and salt leaching

Soil salinity and salt leaching are a risk for sustainable agricultural production in many irrigated areas. This study was conducted over 3.5 years to determine how replacing the usual winter fallow with a cover crop (CC) affects soil salt accumulation and salt leaching in irrigated systems. Treatments studied during the period between summer crops were: barley (Hordeum vulgare L.), vetch (Vicia villosa L.) and fallow. Soil water content was monitored daily to a depth of 1.3 m and used with the numerical model WAVE to calculate drainage. Electrical conductivity (EC) was measured in soil solutions periodically, and in the soil saturated paste extracts before sowing CC and maize. Salt leaching was calculated multiplying drainage by total dissolved salts in the soil solution, and use to obtain a salt balance. Total salt leaching over the four winter fallow periods was 26 Mg ha−1, whereas less than 18 Mg ha−1 in the presence of a CC. Periods of salt gain occurred more often in the CC than in the fallow. By the end of the experiment, net salt losses occurred in all treatments, owing to occasional periods of heavy rainfall. The CC were more prone than the fallow to reduce soil salt accumulation during the early growth stages of the subsequent cash crop.

Bioavailability and extraction of heavy metals from contaminated soil by Atriplex halimus

Pot experiments were performed to evaluate the phytoremediation capacity of plants of Atriplex halimus grown in contaminated mine soils and to investigate the effects of organic amendments on the metal bioavailability and uptake of these metals by plants. Soil samples collected from abandoned mine sites north of Madrid (Spain) were mixed with 0, 30 and 60 Mg ha?1 of two organic amendments, with different pH and nutrients content: pine-bark compost and horse- and sheep-manure compost. The increasing soil organic matter content and pH by the application of manure amendment reduced metal bioavailability in soil stabilising them. The proportion of Cu in the most bioavailable fractions (sum of the water-soluble, exchangeable, acid-soluble and Fe?Mn oxides fractions) decreased with the addition of 60 Mg ha?1 of manure from 62% to 52% in one of the soils studied and from 50% to 30% in the other. This amendment also reduced Zn proportion in water-soluble and exchangeable fractions from 17% to 13% in one of the soils. Manure decreased metal concentrations in shoots of A. halimus, from 97 to 35 mg kg?1 of Cu, from 211 to 98 mg kg?1 of Zn and from 1.4 to 0.6 mg kg?1 of Cd. In these treatments there was a higher plant growth due to the lower metal toxicity and the improvement of nutrients content in soil. This higher growth resulted in a higher total metal accumulation in plant biomass and therefore in a greater amount of metals removed from soil, so manure could be useful for phytoextraction purposes. This amendment increased metal accumulation in shoots from 37 to 138 mg pot?1 of Cu, from 299 to 445 mg pot?1 of Zn and from 1.8 to 3.7 mg pot?1 of Cd. Pine bark amendment did not significantly alter metal availability and its uptake by plants. Plants of A. halimus managed to reduce total Zn concentration in one of the soils from 146 to 130 mg kg?1, but its phytoextraction capacity was insufficient to remediate contaminated soils in the short-to-medium term. However, A. halimus could be, in combination with manure amendment, appropriate for the phytostabilization of metals in mine soils.

Effect of type of dietary carbohydrates on digestion, intestinal barrier and growth performance in 25-d weaned rabbits

Los objetivos globales de esta tesis han sido estudiar el efecto que los carbohidratos de la dieta ejercen sobre los rendimientos productivos, la barrera intestinal, y la digestión de animales destetados a 25 días de edad. Además se ha estudiado cuál es el mejor periodo para determinar la digestibilidad fecal tras el destete a esta edad. En el primer experimento se estudió el efecto de la fibra neutro detergente soluble (FNDS) sobre la barrera intestinal, digestión, microbiota intestinal y rendimientos productivos de gazapos en gazapos en la fase post-destete. Se diseñaron tres piensos isonutritivos en los que la única fuente de variación fueron los niveles de fibra soluble. A partir de una dieta control (AH) con 103 g/kg de materia seca de FNDS y alfalfa como fuente principal de fibra, se sustituyó la mitad de esta alfalfa por una mezcla de pulpa de remolacha y pulpa de manzana (75:25) en el pienso B-AP y por una mezcla de cascarilla y concentrado de proteína de soja (88:12) en el pienso OH, obteniéndose 131 y 79 g/kg de FNDS sobre materia seca, respectivamente. Los conejos se destetaron a 25 días y fueron alimentados con los piensos experimentales hasta los 35 días de edad, momento en el que se sacrificaron para la determinación de la digestibilidad ileal aparente (DIA) de la materia seca (MS), proteína bruta (PB) y almidón, la morfología de la mucosa, y actividad enzimática en el yeyuno, el tejido linfoide asociado a la mucosa, así como la microbiota intestinal. Para la determinación de la morfología de la mucosa se utilizaron adicionalmente 19 animales lactantes de 35 días de edad. Para el estudio de la tasa de mortalidad, se utilizaron 118 animales más por tratamiento que recibieron los piensos experimentales durante las dos semanas post-destete y posteriormente un pienso comercial hasta los 60 días de edad. Los animales recibieron durante todo el experimento medicación en el agua de bebida (100 ppm de apramicina sulfato y 120 ppm de tilosina tartrato). El nivel de fibra soluble mejoró los parámetros que se utilizaron para la caracterización del estado de la barrera intestinal. Los conejos alimentados con el mayor nivel de FNDS en el pienso presentaron una mayor longitud de los villi (P=0.001), un mayor ratio longitud villi/profundidad de las criptas (8.14; P=0.001), una mayor actividad disacaridásica (8671 μmol de glucosa/g de proteína; P=0.019), así como una mayor digestibilidad ileal (96.8%; P=0.002), observándose una reducción en el flujo ileal de almidón a medida que se incrementó el nivel de fibra soluble en el pienso (1,2 vs 0,5 g/d; P=0.001). Los animales lactantes a 35 días de edad presentaron un ratio longitud de villi/profundidad de las criptas menor que el observado en aquéllos alimentados con el pienso B-AP (6.70), pero superior al de los piensos AH y OH. Niveles inferiores de NDFS tendieron (P=0.074) a incrementar la respuesta inmune de tipo celular (linfocitos CD8+). El pienso también afectó a la producción de IL2 (CD25+; P=0.029; CD5+CD25+; P=0.057), pero sin llegar a establecerse una clara relación con el nivel de fibra soluble. La diversidad de la microbiota intestinal no se vio afectada por el pienso (P ≥ 0.38). Los animales alimentados con las piensos B-AP y AH presentaron una reducción en la frecuencia de detección de Clostridium perfringens tanto en íleon (P=0.062) como en ciego (4.3 vs. 17.6%, P =0.047), comparado con el pienso OH. Además la tasa de mortalidad (118 gazapos/pienso) disminuyó de 14.4% en el pienso OH a 5.1% en el pienso B-AP. Entre los 32 y los 35 días de edad se determinó la digestibilidad fecal aparente (14/pienso) de la materia seca (MS), energía bruta (EB), proteína bruta (PB), fibra neutro detergente (FND), fibra ácido detergente (FAD) y almidón. Este grupo, junto con otros nueve animales por tratamiento se utilizaron para determinar el peso del estómago y el ciego, la concentración cecal de ácidos grasos volátiles (AGV) y amoniaco (NH3), así como las tasas de similitud de la microbiota intestinal. Además se estudiaron los rendimientos productivos (35 animales/tratamiento) de los gazapos durante todo el período de cebo, consumiendo los piensos experimentales desde el destete hasta los 35 días y posteriormente un pienso comercial hasta los 60 días de edad. Niveles crecientes de FNDS mejoraron la digestibilidad fecal de la materia seca (MS) y energía (P<0.001). La inclusión FNDS aumentó de manera lineal el peso del contenido cecal (P=0.001) y el peso del aparato digestivo completo (P=0.008), y en los días previos al sacrificio disminuyó de manera lineal el consumo medio diario (P=0.040). Se observó además, una disminución lineal (P≤0.041) del pH del estómago. No se encontró relación entre el pH, la concentración y proporciones molares de AGV y el nivel de FNDS. El pienso pareció tener un efecto, incluso superior al de la madre, sobre la tasa de similitud de la microbiota, y los efectos fueron mayores a nivel cecal que ileal. La eficacia alimenticia aumentó de manera lineal en un 12% entre piensos extremos tras el destete (25- 39d) y en un 3% en el período global de cebo con niveles mayores de NDFS. El consumo medio diario durante la fase post-destete y durante todo el período de cebo, tendió a aumen tar (P≤0.079) con niveles mayores de FNDS, sin embargo no se apreció efecto sobre la ganancia media diaria (P≥0.15). En conclusión, el incremento del nivel de fibra soluble en el pienso parece resultar beneficioso para la salud del animal ya que mejora la integridad de la mucosa, y reduce la frecuencia de detección de potenciales patógenos como C. perfringens y Campylobacter spp. Conforme a estos resultados, debería tenerse en cuenta el contenido en fibra soluble en la formulación de piensos de conejos en la fase post-destete. El objetivo del segundo experimento fue determinar el efecto de la fuente de almidón sobre la digestión, la microbiota intestinal y los rendimientos productivos en conejos destetados con 25 días de edad. Se formularon tres piensos isonutritivos en los que se modificaron las principales fuentes de almidón: trigo crudo, trigo cocido y una combinación de trigo y arroz cocido. Dos grupos de 99 y 193 animales se destetaron con 25 días de edad. El primero de ellos se utilizó para la determinación de los parámetros productivos conforme al mismo protocolo seguido en el experimento anterior. El segundo de los grupos se utilizó para la determinación de la digestibilidad fecal de 32 a 35 d, la digestibilidad ileal aparente (DIA) a 35 d, la morfología de la mucosa intestinal, los parámetros de fermentación cecal; así como, la caracterización de la microbiota intestinal. Se utilizaron además dos grupos adicionales de animales 384 (medicados) y 177 (no medicados) para estudiar el efecto de la suplementación con antibióticos en el agua de bebida sobre la mortalidad. El procesado térmico del trigo mejoró ligeramente la digestibilidad ileal del almidón (P=0.020) pero no modificó el flujo final de almidón que alcanzó el ciego, observándose una mayor frecuencia de detección de Campylobacter spp. y Ruminococcus spp. en ciego (P≤0.023), pero sin cambios a nivel ileal. El procesado térmico del trigo no afectó tampoco a los parámetros productivos, la mortalidad, la digestibilidad ileal y fecal o la morfología de la mucosa. La sustitución parcial del trigo cocido por arroz cocido, penalizó la digestibilidad ileal del almidón (P=0.020) e incrementó el flujo ileal de este nutriente al ciego (P=0.007). Sin embargo no afectó a la mortalidad, pese a que se detectaron cambios en la microbiota tanto a nivel ileal como cecal, disminuyendo la frecuencia de detección de Campylobacter spp. (en íleon y ciego), Helicobacter spp. (en íleon) y Ruminococcus spp (en ciego) e incrementando Bacteroides spp. (en ciego) (P≤0.046). El empleo de arroz cocido en las piensos post-destete no tuvieron efectos sobre los parámetros productivos, la mortalidad, la digestibilidad ileal y fecal a excepción del almidón, o la morfología de la mucosa. La suplementación con antibiótico redujo la fre cuencia de detección de la mayoría de las bacterias estudiadas (P≤0.048), sobre todo para Campylobacter spp., Clostridium perfringens y Propionibacterium spp. (P≤0.048), observándose un efecto mayor a nivel ileal que cecal, lo que se asoció a la bajada significativa (P<0.001) de la mortalidad. En conclusión, los resultados de este experimento indican que la fuente de almidón afecta a la microbiota intestinal pero no influiye sobre la salud del animal. En relación al procesado, el uso de trigo cocido junto con arroz cocido no mejora los resultados obtenidos con trigo duro, si bienserían necesarios más experimentos que confirmaran este punto. El último de los experimentos se centró en un aspecto metodológico. Dado que, los conejos destetados presentan un patrón digestivo diferente al de un animal adulto resultado de su inmadurez digestiva, el objetivo buscado era tratar de determinar el mejor procedimiento para la determinación de la digestibilidad fecal en los gazapos en la fase post-destete. Para tal fin se utilizaron 15 animales/tratamiento de tres camadas diferentes que se destetaron con 25 días, suministrándoles un pienso comercial de crecimiento-cebo. Se registró el consumo medio diario y la excreción diaria de heces desde el día 25 hasta el día 40 de edad para la determinación de la digestibilidad de la MS. La camada afectó al consumo medio diario y la excreción de heces (P=0.013 y 0.014, respectivamente), observándose una tendencia (P=0.061) en la digestibilidad. La edad afectó (P<0.001) a todos estos factores, incrementándose de manera más evidente la excreción que la ingestión de materia seca en la primera semana de vida, para aumentar de forma paralela a partir de la segunda. La correlación entre el consumo medio diario fue mayor con la excreción de heces del mismo día que con la del día siguiente, por lo que se utilizó el primero para la determinación de la digestibilidad de la MS (MSd). La MSd disminuyó de manera lineal hasta los 32 días de edad (2.17±0.25 unidades porcentuales por día), mientras que permaneció constante desde los 32 a los 40 días (69.4±0.47%). Por otro lado, la desviación estándar de la MSd se redujo cuando se incrementó el período de recogida de 2 a 6 días en un 54%. Conforme a los resultados obtenidos, se puede concluir que no es aconsejable comenzar las pruebas de digestibilidad antes de los 32 días de edad y que el número de animales necesario para detectar diferencias significativas entre tratamientos dependerá del período de recogida de heces. ABSTRACT The global aim of this thesis has been to study the effect of dietary carbohydrates on growth, performance, digestion and intestinal barrier in 25-d weaned rabbits. In addition there has also been studied which is the best period to determine the fecal digestibility after weaning. The first experiment focused on the effect of Neutral Detergent Soluble Fibre (NDSF) on gut barrier function, digestion, intestinal microbiota and growth performance n rabbits in the post-weaning period. Three isonutritive diets which only varied in the levels of soluble fiber were formulated such as it described as follows: a control diet (AH) containing 103 g of neutral detergent soluble fiber, including alfalfa as main source of fiber, was replaced by a mixture of beet and apple pulp (75-25) in the B-AP diet and, by a mix of oat hulls and soybean protein concentrate (88:12) in the OH diet, resulting 131 and 79 g of NDFS/kg of dry matter, respectively. Rabbits, weaned at 25 days of age, were fed the experimental diets up to 35 days of age, moment in which they were slaughtered for apparent ileal digestibility (AID) of dry matter (DM), crude protein (CP) and starch, mucosa morphology, sucrose activity, characterization of lamina propria lymphocytes and intestinal microbiota. To assess mucosal morphology, 19 suckling 35-d-old rabbits were also used. For mortality study, besides these animals, 118 additional rabbits per treatment were fed the experimental diets for two weeks period and thereafter received a commercial diet until 60 days of age. Rabbits were water medicated during the whole experimental period (100 ppm de apramicine sulphate and 120 ppm of tylosine tartrate). Level of soluble fiber improved all the parameters used for the characterization of the intestinal barrier condition. Villous height of the jejunal mucosa increased with dietary soluble fiber (P=0.001). Villous height of jejunal mucosa increased with dietary soluble fiber (P = 0.001). Rabbits fed the highest level of soluble fiber (BA-P diet) showed the highest villous height/crypth depth ratio (8.14; P = 0.001), sucrase specific activity (8671 μmol glucose/ g protein; P = 0.019), and the greatest ileal starch digestibility (96.8%; P = 0.002). The opposite effects were observed in rabbits fed decreased levels of soluble fiber (AH and OH diets; 4.70, 5,848 μmol of glucose/g of protein, as average, respectively). The lowest ileal starch digestibility was detected for animal fed OH diet (93.2%). Suckling rabbits of the same age showed a lower villous height/crypt depth ratio (6.70) compared with the B-AP diet group, but this ration was higher that the AH or OH diet groups. Lower levels of soluble fiber tended (P = 0.074) to increase the cellular immune response (CD8+ lymphocytes). Diet affected IL-2 production (CD25+, P = 0.029; CD5+CD25+, P = 0.057), with no clear relationship between soluble fiber and IL-2. The intestinal microbiota biodiversity was not affected by diets (P ≥ 0.38). Animals fed B-AP and AH diets had a reduced cecal frequency of detection compatible with Campylobacter spp. (20.3 vs. 37.8, P = 0.074), and Clostridium perfringens (4.3 vs. 17.6%, P = 0.047), compared with the OH diet group. Moreover, the mortality rates decreased from 14.4 (OH diet) to 5.1% (B-AP diet) with the increased presence of soluble fiber in the diet. Between 32 and 35 days of age, faecal apparent digestibility of dry matter (DM), gross energy (GE), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and starch was determined (14/diet). This group, plus another nine rabbits/diet were used to determine weight of stomach and caecum and their contents, cecal fermentation traits and similarity rate (SR) of intestinal microbiota. Besides, growth performance parameters (35 rabbits/diet) were studied during the whole fattening period, in animals consuming the experimental feed after the weaning up to 35 days of age and later on a commercial diet up animals reached 60 days of age. Increasing levels of Neutral Detergent Soluble Fiber improved faecal dry matter and energy digestibility (P<0.001). NDSF inclusion improved linearly weight of the caecal content (P=0.001) and the total gastrointestinal tract (P=0.008), and in the previous days to slaughter a linear decrease of daily feed intake in diet with highest level of soluble fiber was also observed. Stomach pH decreased linearly with increasing levels of NDFS (P≤0.041). No relation between NDSF level on pH, concentration and molar proportion of VFA was found. Treatments appeared to influence the similarity rate of microbiota, even higher to mother effect. These effects were higher in ileum than in caecum. A linear positive effect of feed efficiency was observed, which increased around 12% in the two weeks post-weaning (25-39d) and 3% in the whole fattening period between extreme diets with highest levels of soluble fiber. Average daily feed intake during the two weeks after weaning and in the whole fattening period, tended (P≤0.079) to increase with highest levels of NDSF; although there were no effect on daily weight gain (≥0.15). In conclusion, an increase of soluble fiber in the feed seems to be beneficial for animal health, due to improve mucose integrity and reduce detection frequency of those poten tial pathogens like C. perfringens and Campylobacter spp. According to these results, level of soluble fiber should be taking care in feed rabbit formulation in the post-weaning period. The objective of the second experiment was to determine the effect of source of starch on digestion, intestinal microbiota and growth performance in twenty-five-day old weaned rabbits. To accomplish with this aim three iso-nutritive diets were formulated with different source of starch: raw wheat, boiled wheat and a combination of boiled wheat and boiled rice. Two groups of 99 and 193 rabbits were weaned at 25 days of age. The first group was used for growth performance determination following the same protocol than in previous experiment. The second group was used to determine faecal digestibility from 32 to 35 d, apparent ileal digestibility (AID) at 35 d, jejunal mucosa morphology, caecal fermentation traits and characterization of intestinal microbiota. For mortality, two additional groups of 384 (medicated) and 177 (not medicated) were used in order to study the effect of antibiotic water supply supplementation. Heat processing of starch slightly improved ileal digestibility of starch (P=0.020) but did not modify the flow of starch to the caecum. An increase in frequency of detection of Campylobacter spp. y Ruminococcus spp. was observed in the caecum (P≤0.023), with no changes at ileal level. Heat processing of wheat did not modify growth performance, mortality, ileal or faecal digestibility and mucosa morphology. Partial substitution of boiled wheat for boiled rice in the diet impaired ileal starch digestibility (P=0.020) and increased the ileal flow of this nutrient to the caecum (P=0.007). However, it did not affect mortality rate, although changes in the ileal and caecal intestinal microbiota were detected, decreasing the frequency of detection of Campylobacter spp. (both ileum and caecum), Helicobacter spp. (at ileum) and Ruminococcus spp (at caecum) and increasing the Bacteroides spp. (at caecum) (P≤0.046). The effect of boiled rice supplementation did not alter growth performance, mortality, ileal or faecal digestibility of other nutrients than starch, and mucosa morphology. Medication of rabbits reduced the ileal frequency of detection of most bacteria studied (P≤0.048), especially for Campylobacter spp., Clostridium perfringens y Propionibacterium spp. (P≤0.048), resulting the effect higher at ileal than caecal level and relating it with a strong reduction of mortality rate (P<0.001). In conclusion, the results of this experiment make think that the source of starch affects the intestinal microbiota but they do not seem to influence animal health. In relation to the effect of heat processed the use of cooked wheat or cooked rice it does not seem to im prove the results obtained with hard wheat, but there would be necessary more experiments that were confirming this point. The last experiment focused on a methodological aspect. Considering that, weaned rabbits have a different digestive pattern than older animals due to their digestive immaturity; the fixed objective was to determine the best procedure for faecal digestibility determination in young rabbits in the post-weaning period. Fifteen rabbits from 5 different litters were weaned at 25 days of age and fed with a commercial feed. Feed intake and faeces excretion were recorded daily from 25 to 40 days of age for dry matter digestibility (DMd) determination. Litter affected daily DM intake and excretion (P=0.013 y 0.014, respectively) and tended to affect DMd (P=0.061). Age affected all these factors (P<0.001), but ingestion increased slowly than dry matter excretion during the first week buth they evolved similarly in the second week. The correlation between daily feed intakes was higher with the faeces excretion of the day than with faeces excretion of the next day, and the first values were used to determine daily DMd. The DMd decreased linearly from weaning to 32 d of age (2.17±0.25 percentage units per day), whereas from 32 to 40 d remained constant (69.4±0.47%). On the other hand, average standard deviation of DMd decreased by 54% when the length of collection period increased from 2 to 6d. Consequently to the obtained results, it could be concluded that it would not be advisable to start digestibility trials before the 32 days of age and that the number of animals required to detect a significant difference among means would depend on the collection period.

Reduction of Nutrient Losses with Eroded Sediments by Post-Fire Soil Stabilisation Techniques

After an experimental fire in steep shrub land in a temperate–humid region (north-west Spain), the effects of two post-fire stabilisation treatments (grass seeding and straw mulching) on the chemical properties of eroded sediments,and the amount of nutrients lost with them, we reevaluated relative to control burnt soil, over a period of 13 months. Total C and N concentrations, and d 13 C, indicated that sediments were mainly contributed by charred plant and litter material. The highest concentrations of extractable base cations in the sediments occurred during the first 3 months following fire, especially for Na and K. As treatments had little or no effect on nutrient concentration in sediments, differences in nutrient losses were due to the 10-fold lower sediment production in mulching compared with other treatments. In control and seeding treatments, the accumulated amounts of nutrients lost with sediments were 989–1028kgha 1 (C), 77kgha 1 (N), 1.9–2.4kgha 1 (Ca), 0.9–1.1kgha 1 (Mg), 0.48–0.55kgha 1 (NH 4 þ –N), 0.39–0.56kgha 1 (K), 0.19–0.34kgha 1 (Na) and , 0.1kgha 1 (P and NO 3 –N) . These values accounted for 22–25% (total C and N) and 5–12% (NH 4 þ –N, Ca, P and Mg) of available nutrients in ash, and 1.0–2.4% of those in ash þ topsoil. As nutrient and sediment losses were strongly correlated, the reduction of the latter by mulching application leads to an effective decrease of post-fire nutrient losses.

Impact of fire and post-fire management techniques on soil chemical properties

The effects of fire ( Control burned soil) and two emergency stabilisation techniques (grass Seeding and straw Mulching ) on 20 chemical characteristics were evaluated on 0 – 5 cm top-soils sampled 1, 90, 180 and 365 days after an experimental fi re in a steep shrubland of a temperate-humid region (NW Spain). Most part of pH (in H 2 O and KCl) variance was explained by the sampling date. No clear temporal trends were identi fi able for total soil C and N content, likely due to the large SOM pool in these soils; however, changes on soil δ 13 C were explained by the deposition of 13 C-depleted ashes, followed by its progressive erosion, while those on soil δ 15 N were a consequence of fi re induced N outputs. After the fi re, NH 4 + – N, P, Na, K, Mg, Ca, Mn, Cu, Zn and B concentrations increased, while those of NO 3 − – N, Al, Fe and Co did not vary significantly. Despite a significant decline with time, concentrations of Mg, Ca and Mn at the end of the study were still higher than in unburned soil, while those of K, Cu, Zn and B were similar to the pre-fire levels and those of NH 4 + – N, P and Na were below pre-fire values. Mulching and Seeding treatments for burned soil emergency stabilisation had significant effects on soil δ 15 N and extractable K, Mg and Ca, while data were inconclusive for their possible effects on the extractable Al, Fe and Co