Abundance and biomass of mesozooplankton from the S-R02 cruise in the western part of the Black Sea in September 2008

The SESAME dataset contains mesozooplankton data collected during September 2008 in the North-West Black Sea (between 44°46' N and 42°29'N latitude and 28°64'E and 30°59'E longitude). Mesozooplankton sampling was undertaken at 9 stations where samples were collected using a Juday net in the 0-10, 10-25, 25-50, 50-100, 100-150, 150-200 m layer. The dataset includes 30 samples analysed for mesozooplankton species composition, species abundance and total biomass. Sampling volume was estimated by multiplying the mouth area with the wire length. The Taxon-specific mesozooplankton abundance sample or aliquots were analyzed under the binocular microscope. Taxonomic identification was done according to Morduhai-Boltovskii et al. 1968. Total biomass was estimated using a tabel with wet weight for each species an stage (Petipa method).

Abundance and biomass of mesozooplankton collected from 1987-1991 allong the romanian littoral

The Longitudinale 1987-1991dataset contains zooplankton data collected from May to October 1987-1991 in 14 station allong 2 transect paralel to the romanian littoral. Zooplankton sampling was undertaken at 14 stations where samples were collected using a Juday closing net in the 0-10, 10-20, 20-30 and 30-40 layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance and biomass of zooplankton collected monthly from January 1978 to december 1978 in front of Constanta city, Black Sea

The Est Constanta 1978 dataset contains zooplankton data collected monthly from January 1978 to december 1978 allong a 5 station transect in front of the city Constanta (44°10'N, 28°41.5'E - EC1; 44°10'N, 28°47'E - EC2; 44°10'N, 28°54'E - EC3; 44°10'N, 29°08'E - EC4; 44°10'N, 29°22'E - EC5). Zooplankton sampling was undertaken at 5 stations where samples were collected using a Juday closing net in the 0-10, 10-25, 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance and biomass of mesozooplankton collected in front of the Danube Delta in April and September 1977

The Gurile Dunarii 1977 dataset contains zooplankton data collected in April and September 1977 in 14 station allong 3 transect in front of the Danube Delta. Zooplankton sampling was undertaken at 14 stations where samples were collected using a Juday closing net in the 0-10, 10-20, 20-30, 30-40 and 40-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance and biomass of mesozooplankton from the Romanian littoral during DANUBS_2001 in spring and autumn 2001

The Danubs 2001 dataset contains zooplankton data collected in March, June, September and October 2001 in 11 station allong 5 transect in front of the Romanian littoral. Zooplankton sampling was undertaken at 11 stations where samples were collected using a Juday closing net in the 0-10, 10-25, and 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance and biomass of zooplankton from the Romanian littoral collected in May, July and September 1998

The SHELF 1998 dataset contains zooplankton data collected in May, July and September 19978 allong 5 transect in front of the Romanian littoral. Zooplankton sampling was undertaken using a Juday closing net in the 0-10, 10-25, and 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance and biomass of zooplankton collected monthly from January 1979 to december 1979 in front of Constanta city, Black Sea

The Est Constanta 1979 dataset contains zooplankton data collected monthly from January 1979 to december 1979 allong a 5 station transect in front of the city Constanta (44°10'N, 28°41.5'E - EC1; 44°10'N, 28°47'E - EC2; 44°10'N, 28°54'E - EC3; 44°10'N, 29°08'E - EC4; 44°10'N, 29°22'E - EC5). Zooplankton sampling was undertaken at 5 stations where samples were collected using a Juday closing net in the 0-10, 10-25, 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Abundance and biomass of zooplankton from the Romanian littoral collected in April, June and September 1999

The SHELF 1999 dataset contains zooplankton data collected in April, June and September 1999 allong 5 transect in front of the Romanian littoral. Zooplankton sampling was undertaken using a Juday closing net in the 0-10, 10-25, and 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Evaluation of cover crops based on characteristics of nitrogen use, and aerial and root growth

La caracterización de los cultivos cubierta (cover crops) puede permitir comparar la idoneidad de diferentes especies para proporcionar servicios ecológicos como el control de la erosión, el reciclado de nutrientes o la producción de forrajes. En este trabajo se estudiaron bajo condiciones de campo diferentes técnicas para caracterizar el dosel vegetal con objeto de establecer una metodología para medir y comparar las arquitecturas de los cultivos cubierta más comunes. Se estableció un ensayo de campo en Madrid (España central) para determinar la relación entre el índice de área foliar (LAI) y la cobertura del suelo (GC) para un cultivo de gramínea, uno de leguminosa y uno de crucífera. Para ello se sembraron doce parcelas con cebada (Hordeum vulgare L.), veza (Vicia sativa L.), y colza (Brassica napus L.). En 10 fechas de muestreo se midieron el LAI (con estimaciones directas y del LAI-2000), la fracción interceptada de la radiación fotosintéticamente activa (FIPAR) y la GC. Un experimento de campo de dos años (Octubre-Abril) se estableció en la misma localización para evaluar diferentes especies (Hordeum vulgare L., Secale cereale L., x Triticosecale Whim, Sinapis alba L., Vicia sativa L.) y cultivares (20) en relación con su idoneidad para ser usadas como cultivos cubierta. La GC se monitorizó mediante análisis de imágenes digitales con 21 y 22 muestreos, y la biomasa se midió 8 y 10 veces, respectivamente para cada año. Un modelo de Gompertz caracterizó la cobertura del suelo hasta el decaimiento observado tras las heladas, mientras que la biomasa se ajustó a ecuaciones de Gompertz, logísticas y lineales-exponenciales. Al final del experimento se determinaron el C, el N y el contenido en fibra (neutrodetergente, ácidodetergente y lignina), así como el N fijado por las leguminosas. Se aplicó el análisis de decisión multicriterio (MCDA) con objeto de obtener un ranking de especies y cultivares de acuerdo con su idoneidad para actuar como cultivos cubierta en cuatro modalidades diferentes: cultivo de cobertura, cultivo captura, abono verde y forraje. Las asociaciones de cultivos leguminosas con no leguminosas pueden afectar al crecimiento radicular y a la absorción de N de ambos componentes de la mezcla. El conocimiento de cómo los sistemas radiculares específicos afectan al crecimiento individual de las especies es útil para entender las interacciones en las asociaciones, así como para planificar estrategias de cultivos cubierta. En un tercer ensayo se combinaron estudios en rhizotrones con extracción de raíces e identificación de especies por microscopía, así como con estudios de crecimiento, absorción de N y 15N en capas profundas del suelo. Las interacciones entre raíces en su crecimiento y en el aprovisionamiento de N se estudiaron para dos de los cultivares mejor valorados en el estudio previo: uno de cebada (Hordeum vulgare L. cv. Hispanic) y otro de veza (Vicia sativa L. cv. Aitana). Se añadió N en dosis de 0 (N0), 50 (N1) y 150 (N2) kg N ha-1. Como resultados del primer estudio, se ajustaron correctamente modelos lineales y cuadráticos a la relación entre la GC y el LAI para todos los cultivos, pero en la gramínea alcanzaron una meseta para un LAI>4. Antes de alcanzar la cobertura total, la pendiente de la relación lineal entre ambas variables se situó en un rango entre 0.025 y 0.030. Las lecturas del LAI-2000 estuvieron correlacionadas linealmente con el LAI, aunque con tendencia a la sobreestimación. Las correcciones basadas en el efecto de aglutinación redujeron el error cuadrático medio del LAI estimado por el LAI-2000 desde 1.2 hasta 0.5 para la crucífera y la leguminosa, no siendo efectivas para la cebada. Esto determinó que para los siguientes estudios se midieran únicamente la GC y la biomasa. En el segundo experimento, las gramíneas alcanzaron la mayor cobertura del suelo (83-99%) y la mayor biomasa (1226-1928 g m-2) al final del mismo. Con la mayor relación C/N (27-39) y contenido en fibra digestible (53-60%) y la menor calidad de residuo (~68%). La mostaza presentó elevadas GC, biomasa y absorción de N en el año más templado en similitud con las gramíneas, aunque escasa calidad como forraje en ambos años. La veza presentó la menor absorción de N (2.4-0.7 g N m-2) debido a la fijación de N (9.8-1.6 g N m-2) y escasa acumulación de N. El tiempo térmico hasta alcanzar el 30% de GC constituyó un buen indicador de especies de rápida cubrición. La cuantificación de las variables permitió hallar variabilidad entre las especies y proporcionó información para posteriores decisiones sobre la selección y manejo de los cultivos cubierta. La agregación de dichas variables a través de funciones de utilidad permitió confeccionar rankings de especies y cultivares para cada uso. Las gramíneas fueron las más indicadas para los usos de cultivo de cobertura, cultivo captura y forraje, mientras que las vezas fueron las mejor como abono verde. La mostaza alcanzó altos valores como cultivo de cobertura y captura en el primer año, pero el segundo decayó debido a su pobre actuación en los inviernos fríos. Hispanic fue el mejor cultivar de cebada como cultivo de cobertura y captura, mientras que Albacete como forraje. El triticale Titania alcanzó la posición más alta como cultiva de cobertura, captura y forraje. Las vezas Aitana y BGE014897 mostraron buenas aptitudes como abono verde y cultivo captura. El MCDA permitió la comparación entre especies y cultivares proporcionando información relevante para la selección y manejo de cultivos cubierta. En el estudio en rhizotrones tanto la mezcla de especies como la cebada alcanzaron mayor intensidad de raíces (RI) y profundidad (RD) que la veza, con valores alrededor de 150 cruces m-1 y 1.4 m respectivamente, comparados con 50 cruces m-1 y 0.9 m para la veza. En las capas más profundas del suelo, la asociación de cultivos mostró valores de RI ligeramente mayores que la cebada en monocultivo. La cebada y la asociación obtuvieron mayores valores de densidad de raíces (RLD) (200-600 m m-3) que la veza (25-130) entre 0.8 y 1.2 m de profundidad. Los niveles de N no mostraron efectos claros en RI, RD ó RLD, sin embargo, el incremento de N favoreció la proliferación de raíces de veza en la asociación en capas profundas del suelo, con un ratio cebada/veza situado entre 25 a N0 y 5 a N2. La absorción de N de la cebada se incrementó en la asociación a expensas de la veza (de ~100 a 200 mg planta-1). Las raíces de cebada en la asociación absorbieron también más nitrógeno marcado de las capas profundas del suelo (0.6 mg 15N planta-1) que en el monocultivo (0.3 mg 15N planta-1). ABSTRACT Cover crop characterization may allow comparing the suitability of different species to provide ecological services such as erosion control, nutrient recycling or fodder production. Different techniques to characterize plant canopy were studied under field conditions in order to establish a methodology for measuring and comparing cover crops canopies. A field trial was established in Madrid (central Spain) to determine the relationship between leaf area index (LAI) and ground cover (GC) in a grass, a legume and a crucifer crop. Twelve plots were sown with either barley (Hordeum vulgare L.), vetch (Vicia sativa L.), or rape (Brassica napus L.). On 10 sampling dates the LAI (both direct and LAI-2000 estimations), fraction intercepted of photosynthetically active radiation (FIPAR) and GC were measured. A two-year field experiment (October-April) was established in the same location to evaluate different species (Hordeum vulgare L., Secale cereale L., x Triticosecale Whim, Sinapis alba L., Vicia sativa L.) and cultivars (20) according to their suitability to be used as cover crops. GC was monitored through digital image analysis with 21 and 22 samples, and biomass measured 8 and 10 times, respectively for each season. A Gompertz model characterized ground cover until the decay observed after frosts, while biomass was fitted to Gompertz, logistic and linear-exponential equations. At the end of the experiment C, N, and fiber (neutral detergent, acid and lignin) contents, and the N fixed by the legumes were determined. Multicriteria decision analysis (MCDA) was applied in order to rank the species and cultivars according to their suitability to perform as cover crops in four different modalities: cover crop, catch crop, green manure and fodder. Intercropping legumes and non-legumes may affect the root growth and N uptake of both components in the mixture. The knowledge of how specific root systems affect the growth of the individual species is useful for understanding the interactions in intercrops as well as for planning cover cropping strategies. In a third trial rhizotron studies were combined with root extraction and species identification by microscopy and with studies of growth, N uptake and 15N uptake from deeper soil layers. The root interactions of root growth and N foraging were studied for two of the best ranked cultivars in the previous study: a barley (Hordeum vulgare L. cv. Hispanic) and a vetch (Vicia sativa L. cv. Aitana). N was added at 0 (N0), 50 (N1) and 150 (N2) kg N ha-1. As a result, linear and quadratic models fitted to the relationship between the GC and LAI for all of the crops, but they reached a plateau in the grass when the LAI > 4. Before reaching full cover, the slope of the linear relationship between both variables was within the range of 0.025 to 0.030. The LAI-2000 readings were linearly correlated with the LAI but they tended to overestimation. Corrections based on the clumping effect reduced the root mean square error of the estimated LAI from the LAI-2000 readings from 1.2 to less than 0.50 for the crucifer and the legume, but were not effective for barley. This determined that in the following studies only the GC and biomass were measured. In the second experiment, the grasses reached the highest ground cover (83- 99%) and biomass (1226-1928 g/m2) at the end of the experiment. The grasses had the highest C/N ratio (27-39) and dietary fiber (53-60%) and the lowest residue quality (~68%). The mustard presented high GC, biomass and N uptake in the warmer year with similarity to grasses, but low fodder capability in both years. The vetch presented the lowest N uptake (2.4-0.7 g N/m2) due to N fixation (9.8-1.6 g N/m2) and low biomass accumulation. The thermal time until reaching 30% ground cover was a good indicator of early coverage species. Variable quantification allowed finding variability among the species and provided information for further decisions involving cover crops selection and management. Aggregation of these variables through utility functions allowed ranking species and cultivars for each usage. Grasses were the most suitable for the cover crop, catch crop and fodder uses, while the vetches were the best as green manures. The mustard attained high ranks as cover and catch crop the first season, but the second decayed due to low performance in cold winters. Hispanic was the most suitable barley cultivar as cover and catch crop, and Albacete as fodder. The triticale Titania attained the highest rank as cover and catch crop and fodder. Vetches Aitana and BGE014897 showed good aptitudes as green manures and catch crops. MCDA allowed comparison among species and cultivars and might provide relevant information for cover crops selection and management. In the rhizotron study the intercrop and the barley attained slightly higher root intensity (RI) and root depth (RD) than the vetch, with values around 150 crosses m-1 and 1.4 m respectively, compared to 50 crosses m-1 and 0.9 m for the vetch. At deep soil layers, intercropping showed slightly larger RI values compared to the sole cropped barley. The barley and the intercropping had larger root length density (RLD) values (200-600 m m-3) than the vetch (25-130) at 0.8-1.2 m depth. The topsoil N supply did not show a clear effect on the RI, RD or RLD; however increasing topsoil N favored the proliferation of vetch roots in the intercropping at deep soil layers, with the barley/vetch root ratio ranging from 25 at N0 to 5 at N2. The N uptake of the barley was enhanced in the intercropping at the expense of the vetch (from ~100 mg plant-1 to 200). The intercropped barley roots took up more labeled nitrogen (0.6 mg 15N plant-1) than the sole-cropped barley roots (0.3 mg 15N plant-1) from deep layers.

Potenciación de la respuesta productiva en rumiantes mediante el empleo de proteínas protegidas

El principal objetivo de esta tesis fue incrementar la eficiencia proteica en las dietas de rumiantes mediante el uso de proteínas protegidas (harina de girasol y guisante de primavera), así como mejorar la predicción de los aportes de proteína microbiana. Una partida de harinas comerciales de girasol (HG) y de guisante de primavera (GP) fueron tratadas con soluciones 4 N de ácido málico (268,2 g/L) o ácido ortofosfórico (130,6 g/L). Para cada harina, ácido y día de tratamiento, dos fracciones de 12,5 kg fueron pulverizadas sucesivamente en una hormigonera con la solución de ácido correspondiente mediante un pulverizador de campo. Las dos fracciones fueron mezcladas posteriormente y se dejaron reposar durante 1 h a temperatura ambiente. La mezcla fue luego secada en una estufa de aire forzado a 120 ºC durante 1 h. La estufa fue apagada inmediatamente después y el material tratado se mantuvo dentro de ésta hasta la mañana siguiente. El material fue removido durante el proceso de secado cada 30 min durante las primeras 2 h y cada 60 min durante las 5 h posteriores. Este proceso se repitió hasta conseguir las cantidades de harinas tratadas necesarias en los distintos ensayos. En el primer experimento (capitulo 3) se llevaron a cabo estudios de digestión ruminal e intestinal para evaluar los efectos de la aplicación de las soluciones ácidas indicadas y calor a fin de proteger las proteínas de HG y GP contra la degradación ruminal. Estos estudios se realizaron con tres corderos canulados en el rumen y en el duodeno. El estudio de digestión ruminal fue realizado en tres periodos experimentales en los que los corderos fueron alimentados sucesivamente con tres dietas isoproteicas que incluían HG y GP, sin tratar o tratadas con ácidos málico u ortofosfórico. Cada periodo experimental de 21 días incluyó sucesivamente: 10 días de adaptación a las dietas, un estudio del tránsito ruminal de las partículas de HG y GP (días 11 a 14), y la incubación de las muestras de ambos alimentos en bolsas de nailon (días 15–21). Las harinas incubadas en cada periodo experimental correspondieron a las que fueron incluidas en las dietas. Las bacterias ruminales fueron marcadas desde el día 11 hasta el día 21 del periodo experimental mediante infusión intra-ruminal continua con una fuente de 15N. Tras finalizar las incubaciones in situ el día 21 el rumen fue vaciado en cada periodo para aislar las bacterias asociadas a la fase sólida y liquida del rumen. El estudio de digestión intestinal fue realizado veinte días después del final del estudio ruminal a fin de eliminar el enriquecimiento en 15N de la digesta. En este estudio se incubaron muestras compuestas obtenidas mediante la combinación de los diferentes residuos no degradados en el rumen de forma que fuesen representativas de la composición química de la fracción no degradada en el rumen (RU). En esta fase los corderos fueron alimentados con la dieta sin tratar para determinar la digestibilidad de las harinas tanto tratadas como sin tratar mediante la técnica de las bolsas móviles. Además, las proteínas contenidas en las harinas tratadas y sin tratar, así como en las muestras correspondientes a los residuos a 0 h, las muestras compuestas anteriormente indicadas y las muestras no digeridas intestinalmente fueron extraídas y sometidas a electroforesis para determinar el sitio de digestión de las diferentes fracciones proteicas. Las estimaciones de la RU y la digestibilidad intestinal de la materia seca, la materia orgánica (solamente para RU), la proteína bruta (PB) y el almidón (solamente en GP) fueron obtenidos considerando la contaminación microbiana y las tasas de conminución y salida de partículas. Las estimaciones de RU y de la digestibilidad intestinal disminuyeron en todas las fracciones evaluadas de ambos alimentos al corregir por la contaminación microbiana acaecida en el rumen. Todas las estimaciones de RU aumentaron con los tratamientos de protección, incrementándose también la digestibilidad intestinal de la materia seca en la HG. Los bajos valores de la digestibilidad de la proteína de GP tratado y sin tratar sugieren la presencia de algún factor antitripsico no termolábil es esta harina. Los tratamientos de protección incrementaron consistentemente la fracción de materia seca y PB digerida intestinalmente en los dos alimentos, mientras que la fracción de almidón en la muestra de GP solamente aumentó numéricamente (60,5% de media). Sin embargo, los tratamientos también redujeron la fermentación de la materia orgánica, lo cual podría disminuir la síntesis de proteína microbiana. Los estudios de electroforesis muestran la práctica desaparición de la albumina por la degradación ruminal en ambos alimentos, así como que los cambios en otras proteínas de la muestra RU fueron más pronunciados en GP que en HG. La composición de las bacterias asociadas con las fases de digesta ruminal sólida (BAS) y líquida (BAL) fue estudiada para revisar la precisión de un sistema de predicción previo que determinaba la infravaloración del aporte de nutrientes correspondiente a las BAS cuando de usa 15N como marcador y las BAL como referencia microbiana (capitulo 4). Al comparar con BAS, BAL mostraron menores contenidos en materia orgánica, polisacáridos de glucosa y lípidos totales y un mayor contenido en PB, así como un mayor enriquecimiento en 15N. Los datos obtenidos en el estudio actual se ajustan bien a la ecuación previa que predice el enriquecimiento en 15N de las BAS a partir del mismo valor en BAL. Esta nueva ecuación permite establecer que se produce una infravaloración de un 22% en el aporte de PB al animal a partir de las BAS sintetizadas si las BAL son usadas como muestras de referencia. Una segunda relación calculada utilizando los valores medios por dieta expuestos en numerosos trabajos encontrados en la literatura confirma la magnitud de este error. Esta infravaloración asociada al uso de BAL como referencia fue mayor para el aporte de glucosa (43,1%) y todavía mayor para el aporte de lípidos (59,9%), como consecuencia de los menores contenidos de ambas fracciones en BAL frente a SAB. Estos errores deberían ser considerados para obtener mayor precisión en la estimación del aporte de nutrientes microbianos y mejorar la nutrición de los rumiantes. En el experimento 2 se realizó un estudio de producción (capitulo 5) para evaluar los efectos del tratamiento de las harinas HG y GP con soluciones de ácido málico o ácido ortofosfórico sobre el crecimiento, el consumo de concentrado y el rendimiento y engrasamiento de las canales de corderos de engorde. Noventa corderos machos de cruce entrefino procedentes de tres granjas comerciales (peso inicial medio = 14,6, 15,3 y 13,3 kg, respectivamente) fueron asignados aleatoriamente a cinco dietas con diferentes niveles de proteína y diferentes tratamientos con ácidos y engordados hasta un peso medio al sacrificio de 25 kg. Las fuentes de proteína en el pienso control (C; PB=18,0%) fueron harina de soja, HG y GP sin tratar. En tres de los piensos experimentales, las harinas tratadas con ácido ortofosfórico sustituyeron a las de HG y GP sin tratar (Control Ortofosfórico, PC; PB=18,0% sobre materia seca), sustituyéndose, además, la harina de soja parcialmente (Sustitución Media Ortofosfórico, MSP; PB=16,7%) o totalmente (Sustitución Total Ortofosfórico, TSP; PB=15,6%). Finalmente, en uno de los piensos el ácido ortofosfórico fue reemplazo por acido málico para proteger ambas harinas (Sustitución Media Málico, MSM; PB= 16,7%). La paja de trigo (fuente de forraje) y el concentrado fueron ofrecidos ad libitum. Dieciocho corderos fueron distribuidos en seis cubículos con tres animales para cada dieta. Los datos fueron analizados según un análisis factorial considerando el peso inicial como covariable y la granja de procedencia como bloque. Los datos de consumo de concentrado y eficiencia de conversión fueron analizados usando el cubículo como unidad experimental, mientras que los datos sobre ganancia media diaria, rendimiento a la canal, grasa dorsal y grasa pélvico renal fueron analizados usando el cordero como unidad experimental. No se encontró ningún efecto asociado con el nivel de PB sobre ninguna variable estudiada. Esto sugiere que usando proteínas protegidas es posible utilizar concentrados con 15,6% de PB (sobre materia seca) disminuyendo así la cantidad de concentrados de proteína vegetal a incluir en los piensos y la calidad de los concentrados proteicos. Los corderos alimentados con la dieta MSM tuvieron mayores ganancias medias diarias (15,2%; P= 0,042), y mejores rendimiento a la canal en caliente (1,3 unidades porcentuales; P= 0,037) que los corderos alimentados con el concentrado MSP. Esto podría ser explicado por los efectos benéficos ruminales del malato o por el mayor efecto de protección conseguido con el ácido málico. ABSTRACT The main objective of this thesis project was to increase the protein efficiency in ruminant diets by using protected protein (sunflower meal and spring pea), and improving the prediction of microbial protein supply. Commercial sunflower meal (SFM) and spring pea (SP) were treated with 4 N solutions (200 mL/kg) of malic acid (268.2 g/L) or orthophosphoric acid (130.6 g/L). Daily, two fractions of 12.5 kg of one of these meals were successively sprayed with the tested acid solution in a concrete mixer using a sprayer. Both fractions were then mixed and allowed to rest for 1 h at room temperature. The blend was then dried in a forced air oven at 120 ºC for 1 h. Then the oven was turned off and the treated material was left in the oven overnight. During the drying process, the material was stirred every 30 min during the first 2 h and then every 60 min for the subsequent 5 h. This process was repeated until the amounts of treated flour needed for the different trials performed. In the first experiment (chapter 3), ruminal and intestinal digestion trials were conducted to study the effects of the application of these acid solutions and heat to protect proteins of SFM and SP against ruminal degradation using three wethers fitted with rumen and duodenum cannulae. The ruminal digestion study was carried out in three experimental periods in which the wethers were successively fed three isoproteic diets including SFM and SP, untreated or treated with malic or orthophosphoric acids. The experimental periods of 21 days included successively: 10 days of diet adaptation, SFM and SP particle ruminal transit study (days 11–14) and ruminal nylon-bag incubations (days 15–21). The meals incubated in each experimental period were those corresponding to the associated diet. Rumen bacteria were labelled from days 11 to 21 by continuous intra-ruminal infusion of a 15N source and the rumen was emptied at the end of in situ incubations in each period to isolate solid adherent bacteria and liquid associate bacteria. The intestinal digestion trial was conducted twenty days after the end of the ruminal studies to eliminate the 15N enrichment in the digesta. The tested samples were composite samples obtained pooling the different ruminally undegraded residues to be representative of the chemical composition of the ruminally undegraded fraction (RU). Wethers were fed the untreated diet to determine the intestinal digestibility of untreated and treated meals using the mobile nylon bag technique. In addition, protein in untreated and treated meals and their 0 h, composite and intestinally undigested samples were extracted and subjected to electrophoresis to determine the digestion site of the different protein fractions. Estimates of the RU and its intestinal digestibility of dry matter, organic matter (only for RU), crude protein (CP) and starch (only in SP) were obtained considering ruminal microbial contamination and particle comminution and outflow rates. When corrected for the microbial contamination taking place in the rumen, estimates of RU and intestinal digestibility decreased in all tested fractions for both feeds. All RU estimates increased with the protective treatments, whereas intestinal digestibility-dry matter also increased in SFM. Low intestinal digestibility-CP values in untreated and treated samples suggested the presence of non-heat labile antitrypsin factors in SP. Protective treatments of both feeds led to consistent increases in the intestinal digested fraction of dry matter and CP, being only numerically different for SP-starch (60.5% as average). However, treatments also reduced the organic matter fermentation, which may decrease ruminal microbial protein synthesis. Electrophoretic studies showed albumin disappearance in both SFM and SP, whereas changes in other RU proteins were more pronounced in SP than SFM. The chemical composition of bacteria associated with solid (SAB) and liquid (LAB) rumen-digesta phases was studied to examine the accuracy of a previous regression system determining the underevaluation of SAB-nutrient supply using 15N as marker and LAB as microbial reference (chapter 4). Compared with SAB, LAB showed lower contents of organic matter, polysaccharide-glucose and total lipids and the opposite for the CP content and the 15N enrichment. Present data fitted well to the previous relationship predicting the 15N enrichment of SAB from the same value in LAB. This new equation allows establishing an underevaluation in the supply of CP from the synthesized SAB in 22.0% if LAB is used as reference. Another relationship calculated using mean diet values from the literature confirmed the magnitude of this error. This underevaluation was higher for the supply of glucose (43.1%) and still higher for the lipid supply (59.9%) as a consequence of the lower contents of these both fractions in LAB than in SAB. These errors should be considered to obtain more accurate estimates of the microbial nutrient supply and to improve ruminant nutrition. A production study was performed in experiment 2 (chapter 5) to examine the effects of treating SFM and SP meals with orthophosphoric or malic acid solutions on growth performance, concentrate intake, and carcass yield and fatness of growing-fattening lambs. Ninety "Entrefino" cross male lambs from three commercial farms (average initial body weights (BW) = 14.6, 15.3 and 13.3 kg) were randomly assigned to five diets with different acid treatment and protein levels, and fattened to an average slaughter weight of 25 kg. Protein sources in the control concentrate (C; CP=18%) were soybean meal and untreated SFM and SP. In three of the experimental concentrates, orthophosphoric acid-treated meals substituted untreated SFM and SP (Orthophosphoric Control, PC; CP=18% dry matter basis), and soybean meal was partially (Medium Substitution Orthophosphoric, MSP; CP=16.7%) or totally removed (Total Substitution Orthophosphoric, TSP; CP=15.6%). In addition, in one concentrate orthophosphoric acid was replaced by malic acid to protect these meals (Medium Substitution Malic, MSM; CP= 16.7%). Wheat straw (roughage source) and concentrate were offered ad libitum. Eighteen lambs were allocated to six pens of three animals on each diet. Data were analyzed using a factorial analysis with initial body weight BW as covariate and farm of origin as block. Data on concentrate intake and feed conversion efficiency were analyzed using pen as experimental unit, while data on average daily gain, carcass yield, dorsal fat, and kidney-pelvic-fat were analyzed with lamb as experimental unit. No effect associated with the CP level was observed on any parameter. This suggests that with protected proteins it is possible to feed concentrates with 15.6% CP (dry matter basis) reducing the quantity of vegetable protein meals to include in the concentrate as well as the quality of the protein concentrates. Lambs feed MSM had higher average daily gains (15.2%; P= 0.042), and better hot carcass yields (1.3 percentage points; P= 0.037) than lambs feed MSP. This probably can be explained by ruminal malate actions and by greater protection effects obtained with malic acid.

Multicriteria decision analysis applied to cover crop species andcultivars selection

Cover crop selection should be oriented to the achievement of specific agrosystem benefits. The covercrop, catch crop, green manure and fodder uses were identified as possible targets for selection. Theobjective was to apply multi-criteria decision analysis to evaluate different species (Hordeum vulgareL., Secale cereale L., ×Triticosecale Whim, Sinapis alba L., Vicia sativa L.) and cultivars according to theirsuitability to be used as cover crops in each of the uses. A field trial with 20 cultivars of the five specieswas conducted in Central Spain during two seasons (October?April). Measurements of ground cover, cropbiomass, N uptake, N derived from the atmosphere, C/N, dietary fiber content and residue quality werecollected. Aggregation of these variables through utility functions allowed ranking species and cultivarsfor each usage. Grasses were the most suitable for the cover crop, catch crop and fodder uses, while thevetches were the best as green manures. The mustard attained high ranks as cover and catch crop the firstseason, but the second decayed due to low performance in cold winters. Mustard and vetches obtainedworse rankings than grasses as fodder. Hispanic was the most suitable barley cultivar as cover and catchcrop, and Albacete as fodder. The triticale Titania attained the highest rank as cover and catch crop andfodder. Vetches Aitana and BGE014897 showed good aptitudes as green manures and catch crops. Thisanalysis allowed comparison among species and cultivars and might provide relevant information forcover crops selection and management.

Quantitative characterization of five cover crop species

The introduction of cover crops in the intercrop period may provide a broad range of ecosystem services derived from the multiple functions they can perform, such as erosion control, recycling of nutrients or forage source. However, the achievement of these services in a particular agrosystem is not always required at the same time or to the same degree. Thus, species selection and definition of targeted objectives is critical when growing cover crops. The goal of the current work was to describe the traits that determine the suitability of five species (barley, rye, triticale, mustard and vetch) for cover cropping. A field trial was established during two seasons (October to April) in Madrid (central Spain). Ground cover and biomass were monitored at regular intervals during each growing season. A Gompertz model characterized ground cover until the decay observed after frosts, while biomass was fitted to Gompertz, logistic and linear-exponential equations. At the end of the experiment, carbon (C), nitrogen (N), and fibre (neutral detergent, acid and lignin) contents, and the N fixed by the legume were determined. The grasses reached the highest ground cover (83–99%) and biomass (1226–1928 g/m2) at the end of the experiment. With the highest C:N ratio (27–39) and dietary fibre (527–600 mg/g) and the lowest residue quality (~680 mg/g), grasses were suitable for erosion control, catch crop and fodder. The vetch presented the lowest N uptake (2·4 and 0·7 g N/m2) due to N fixation (9·8 and 1·6 g N/m2) and low biomass accumulation. The mustard presented high N uptake in the warm year and could act as a catch crop, but low fodder capability in both years. The thermal time before reaching 30% ground cover was a good indicator of early coverage species. Variable quantification allowed finding variability among the species and provided information for further decisions involving cover crop selection and management.

The Decisive Step in Betaxanthin Biosynthesis Is a Spontaneous Reaction1

Experiments were performed to confirm that the aldimine bond formation is a spontaneous reaction, because attempts to find an enzyme catalyzing the last decisive step in betaxanthin biosynthesis, the aldimine formation, failed. Feeding different amino acids to betalain-forming hairy root cultures of yellow beet (Beta vulgaris L. subsp. vulgaris “Golden Beet”) showed that all amino acids (S- and R-forms) led to the corresponding betaxanthins. We observed neither an amino acid specificity nor a stereoselectivity in this process. In addition, increasing the endogenous phenylalanine (Phe) level by feeding the Phe ammonia-lyase inhibitor 2-aminoindan 2-phosphonic acid yielded the Phe-derived betaxanthin. Feeding amino acids or 2-aminoindan 2-phosphonic acid to hypocotyls of fodder beet (B. vulgaris L. subsp. vulgaris “Altamo”) plants led to the same results. Furthermore, feeding cyclo-3-(3,4-dihydroxyphenyl)-alanine (cyclo-Dopa) to these hypocotyls resulted in betanidin formation, indicating that the decisive step in betacyanin formation proceeds spontaneously. Finally, feeding betalamic acid to broad bean (Vicia faba L.) seedlings, which are known to accumulate high levels of Dopa but do not synthesize betaxanthins, resulted in the formation of dopaxanthin. These results indicate that the condensation of betalamic acid with amino acids (possibly including cyclo-Dopa or amines) in planta is a spontaneous, not an enzyme-catalyzed reaction.

Primary and secondary metabolites production in signal grass around the year under nitrogen fertilizer

Plants produce a number of substances and products and primary and secondary metabolites (SM) are amongst them with many benefits but limitation as well. Usually, the fodder are not considered toxic to animals or as a source having higher SM. The Brachiaria decumbens has a considerable nutritional value, but it is considered as a toxic grass for causing photosensitization in animals, if the grass is not harvested for more than 30 days or solely. The absence of detailed information in the literature about SM in Brachiaria, metabolites production and its chemical profile enable us to focus not only on the nutritive value but to get answers in all aspects and especially on toxicity. The study was conducted in the period of december 2013 to december 2014; in greenhouse FZEA-USP. B. decumbens was used with two cutting heights (10 and 20 cm) and nitrogen doses (0, 150, 300 and 450 kg ha-1) in complete randomized block design. The bromatological analysis were carried out on near infrared spectroscopy. Generally, the application of 150 kg ha-1 N was sufficient to promote the nutritional value in B. decumbens but above it the nitrogen use efficiency decline significantly. The highest dry matter yield (99.97 g/pot) was observed in autumn and the lowest was in winter (30.20 g/pot). While, as per nitrogen dose the average highest dry matter yield was at 150 kg ha-1 (79.98 g/pot). The highest crude protein was observed in winter (11.88%) and the lowest in autumn (7.78%). By the cutting heights; the 10 cm proved to have high CP (9.51%). In respect of fibrous contents, the highest acid detergent fiber was noted in summer (36.37%) and lowest in winter (30.88%). While the neutral detergent fiber was being highest in autumn and lowest in spring (79.60%). The highest in vitro dry matter and organic matter digestibilities were noted at 300 kg ha-1 N; being 68.06 and 60.57%; respectively; with the lowest observed in without N treatments (62.63% and 57.97), respectively. For determination of the classes, types and concentration of SM in B. decumbens, phytochemical tests, thin layer and liquid chromatography-mass spectrometry and nuclear magnetic resonance analysis were carried out. Height, nitrogen and seasons significantly (P <0.0001) affected the secondary metabolic profile. A new protodioscin isomer (protoneodioscin (25S-)) was identified for first time in B. decumbens and is supposed to be the probable toxicity reason. Its structure was verified by 1D and 2D NMR techniques (1H, 13C) and 1D (COSY-45, edited HSQC, HMBC, H2BC, HSQC -TOCSY, NOESY and 1 H, 1 H, J). All factors influence the metabolic profile significantly (P <0.0001). The lowest phenols were at 300 kg ha-1 while the lowest flavones were at 0 kg ha-1. Season wise the highest phenols occurred in autumn (19.65 mg/g d.wt.) and highest flavones (28.87 mg/g d.wt.) in spring. Seasons effect the saponin production significantly (P <0.0001) and the results showed significant differences in the protodioscin (17.63±4.3 - 22.57±2.2 mg/g d.wt.) and protoneodioscin (23.3±1.2 - 31.07±2.9 mg/g d.wt.) concentrations. The highest protodioscin isomers concentrations were observed in winter and spring and by N doses the highest were noted in 300 kg ha-1. Simply, all factors significantly played their role in varying concentrations of secondary metabolites.