Effects of the inclusion of oat hulls or sugar beet pulp in the diet on gizzard characteristics, apparent ileal digestibility of nutrients, and microbial count in the ceca in 36 day old broilers reared on floor

The effects of the inclusion of oat hulls (OH) and sugar beet pulp (SBP) in the diet on gizzard characteristics, apparent ileal nutrient digestibility (AID), and Clostridium perfringens, Enterobacteriaceae, and Lactobacillus proliferation in the ceca were studied in 36 d?old broilers. There were a control diet with a low CF content (1.61%) and 2 additional diets that resulted from the dilution of this feed with 5% of either OH or SBP.

Apparent digestibility and metabolizable energy content of lipid sources in poultry

A trial was conducted to determine the apparent digestibility (ATTD) and AME content of different lipid sources in 21d-old broilers. There were a control diet based on corn and soybean meal without any supplemental fat and 6 additional diets forming a 3x2 factorial with 3 sources of fat (soy oil, SBO; reconstituted monoglyceride oil, RMG; and reconstituted triglyceride oil, RTG) included in the diet (3 or 6%) at expenses (wt:wt) of the basal diet.

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.

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

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.

A bacterial cysteine protease effector protein interferes with photosynthesis to suppress plant innate immune responses

The bacterial pathogen Pseudomonas syringae pv tomato DC3000 suppresses plant innate immunity with effector proteins injected by a type III secretion system (T3SS). The cysteine protease effector HopN1, which reduces the ability of DC3000 to elicit programmed cell death in non-host tobacco, was found to also suppress the production of defence-associated reactive oxygen species (ROS) and callose when delivered by Pseudomonas fluorescens heterologously expressing a P. syringae T3SS. Purified His 6 -tagged HopN1 was used to identify tomato PsbQ, a member of the oxygen evolving complex of photosystem II (PSII), as an interacting protein. HopN1 localized to chloroplasts and both degraded PsbQ and inhibited PSII activity in chloroplast preparations, whereas a HopN1 D299A non-catalytic mutant lost these abilities. Gene silencing of NtPsbQ in tobacco compromised ROS production and programmed cell death.

Conductancia del mesófilo en especies forestales: implicaciones en la respuesta fotosintética a la disponibilidad de agua y luz

La fotosíntesis es el proceso biológico que permite la producción primaria y, por tanto, la vida en nuestro planeta. La tasa fotosintética viene determinada por la ‘maquinaria’ bioquímica y las resistencias difusivas al paso del CO2 desde la atmósfera hasta su fijación en el interior de los cloroplastos. Históricamente la mayor resistencia difusiva se ha atribuido al cierre estomático, sin embargo ahora sabemos, debido a las mejoras en las técnicas experimentales, que existe también una resistencia grande que se opone a la difusión del CO2 desde los espacios intercelulares a los lugares de carboxilación. Esta resistencia, llamada normalmente por su inversa: la conductancia del mesófilo (gm), puede ser igual o incluso superior a la resistencia debida por el cierre estomático. En la presente tesis doctoral he caracterizado la limitación que ejerce la resistencia del mesófilo a la fijación de CO2 en diversas especies forestales y en distintos momentos de su ciclo biológico. En la fase de regenerado, hemos estudiado tres situaciones ambientales relevantes en el mayor éxito de su supervivencia, que son: el déficit hídrico, su interacción con la irradiancia y el paso del crecimiento en la sombra a mayor irradiancia, como puede suceder tras la apertura de un hueco en el dosel forestal. En la fase de arbolado adulto se ha caracterizado el estado hídrico y el intercambio gaseoso en hojas desarrolladas a distinta irradiancia dentro del dosel vegetal durante tres años contrastados en pluviometría. Para cada tipo de estudio se han empleado las técnicas ecofisiológicas más pertinentes para evaluar el estado hídrico y el intercambio gaseoso. Por su complejidad y la falta de un método que permita su cuantificación directa, la gm ha sido evaluada por los métodos más usados, que son: la discriminación isotópica del carbono 13, el método de la J variable, el método de la J constante y el método de la curvatura. Los resultados más significativos permiten concluir que la limitación relativa a la fotosíntesis por la conductancia estomática, del mesófilo y bioquímica es dependiente de la localización de la hoja en el dosel forestal. Por primera vez se ha documentado que bajo estrés hídrico las hojas desarrolladas a la sombra estuvieron más limitadas por una reducción en la gm, mientras que las hojas desarrolladas a pleno sol estuvieron más limitadas por reducción mayor de la conductancia estomática (gsw). Encontramos buena conexión entre el aparato fotosintético foliar y el sistema hídrico debido al alto grado de correlación entre la conductancia hidráulica foliar aparente y la concentración de CO2 en los cloroplastos en distintas especies forestales. Además, hemos mostrado diferentes pautas de regulación del intercambio gaseoso según las particularidades ecológicas de las especies estudiadas. Tanto en brinzales crecidos de forma natural y en el arbolado adulto como en plántulas cultivadas en el invernadero la ontogenia afectó a las limitaciones de la fotosíntesis producidas por estrés hídrico, resultando que las limitaciones estomáticas fueron dominantes en hojas más jóvenes mientras que las no estomáticas en hojas más maduras. La puesta en luz supuso un gran descenso en la gm durante los días siguientes a la transferencia, siendo este efecto mayor según el grado de sombreo previo en el que se han desarrollado las hojas. La aclimatación de las hojas a la alta irradiancia estuvo ligada a las modificaciones anatómicas foliares y al estado de desarrollo de la hoja. El ratio entre la gm/gsw determinó la mayor eficiencia en el uso del agua y un menor estado oxidativo durante la fase de estrés hídrico y su posterior rehidratación, lo cual sugiere el uso de este ratio en los programas de mejora genética frente al estrés hídrico. Debido a que la mayoría de modelos de estimación de la producción primaria bruta (GPP) de un ecosistema no incluye la gm, los mismos están incurriendo en una sobreestimación del GPP particularmente bajo condiciones de estrés hídrico, porque más de la mitad de la reducción en fotosíntesis en hojas desarrolladas a la sombra se debe a la reducción en gm. Finalmente se presenta un análisis de la importancia en las estimas de la gm bajo estrés hídrico de la refijación del CO2 emitido en la mitocondria a consecuencia de la fotorrespiración y la respiración mitocondrial en luz. ABSTRACT Photosynthesis is the biological process that supports primary production and, therefore, life on our planet. Rates of photosynthesis are determined by biochemical “machinery” and the diffusive resistance to the transfer of CO2 from the atmosphere to the place of fixation within the chloroplasts. Historically the largest diffusive resistance was attributed to the stomata, although we now know via improvements in experimental techniques that there is also a large resistance from sub-stomatal cavities to sites of carboxylation. This resistance, commonly quantified as mesophyll conductance (gm), can be as large or even larger than that due to stomatal resistance. In the present PhD I have characterized the limitation exerted by the mesophyll resistance to CO2 fixation in different forest species at different stages of their life cycle. In seedlings, we studied three environmental conditions that affect plant fitness, namely, water deficit, the interaction of water deficit with irradiance, and the transfer of plants grown in the shade to higher irradiance as can occur when a gap opens in the forest canopy. At the stage of mature trees we characterized water status and gas exchange in leaves developed at different irradiance within the canopy over the course of three years that had contrasting rainfall. For each study we used the most relevant ecophysiological techniques to quantify water relations and gas exchange. Due to its complexity and the lack of a method that allows direct quantification, gm was estimated by the most commonly used methods which are: carbon isotope discrimination, the J-variable, constant J and the curvature method The most significant results suggest that the relative limitation of photosynthesis by stomata, mesophyll and biochemistry depending on the position of the leaf within the canopy. For the first time it was documented that under water stress shaded leaves were more limited by a reduction in gm, while the sun-adapted leaves were more limited by stomatal conductance (gsw). The connection between leaf photosynthetic apparatus and the hydraulic system was shown by the good correlations found between the apparent leaf hydraulic conductance and the CO2 concentration in the chloroplasts in shade- and sun-adapted leaves of several tree species. In addition, we have revealed different patterns of gas exchange regulation according to the functional ecology of the species studied. In field grown trees and greenhouse-grown seedlings ontogeny affected limitations of photosynthesis due to water stress with stomatal limitations dominating in young leaves and nonstomatal limitations in older leaves. The transfer to high light resulted in major decrease of gm during the days following the transfer and this effect was greater as higher was the shade which leaves were developed. Acclimation to high light was linked to the leaf anatomical changes and the state of leaf development. The ratio between the gm/gsw determined the greater efficiency in water use and reduced the oxidative stress during the water stress and subsequent rehydration, suggesting the use of this ratio in breeding programs aiming to increase avoidance of water stress. Because most models to estimate gross primary production (GPP) of an ecosystem do not include gm, they are incurring an overestimation of GPP particularly under conditions of water stress because more than half of An decrease in shade-developed leaves may be due to reduction in gm. Finally, we present an analysis of the importance of how estimates of gm under water stress are affected by the refixation of CO2 that is emitted from mitochondria via photorespiration and mitochondrial respiration in light.

Implications of the mesophyll conductance to CO2 for photosynthesis and water-use efficiency during long-term water stress and recovery in two contrasting Eucalyptu species

Water stress (WS) slows growth and photosynthesis (An), but most knowledge comes from short-time studies that do not account for longer term acclimation processes that are especially relevant in tree species. Using two Eucalyptus species that contrast in drought tolerance, we induced moderate and severe water deficits by withholding water until stomatal conductance (gsw) decreased to two pre-defined values for 24 d, WS was maintained at the target gsw for 29 d and then plants were re-watered. Additionally, we developed new equations to simulate the effect on mesophyll conductance (gm) of accounting for the resistance to refixation of CO2. The diffusive limitations to CO2, dominated by the stomata, were the most important constraints to An. Full recovery of An was reached after re-watering, characterized by quick recovery of gm and even higher biochemical capacity, in contrast to the slower recovery of gsw. The acclimation to long-term WS led to decreased mesophyll and biochemical limitations, in contrast to studies in which stress was imposed more rapidly. Finally, we provide evidence that higher gm under WS contributes to higher intrinsic water-use efficiency (iWUE) and reduces the leaf oxidative stress, highlighting the importance of gm as a target for breeding/genetic engineering.

Initializing the DSSAT-Century model: inverse calibration of carbon pools from apparent soil N mineralization

The CENTURY soil organic matter model was adapted for the DSSAT (Decision Support System for Agrotechnology Transfer), modular format in order to better simulate the dynamics of soil organic nutrient processes (Gijsman et al., 2002). The CENTURY model divides the soil organic carbon (SOC) into three hypothetical pools: microbial or active material (SOC1), intermediate (SOC2) and the largely inert and stable material (SOC3) (Jones et al., 2003). At the beginning of the simulation, CENTURY model needs a value of SOC3 per soil layer which can be estimated by the model (based on soil texture and management history) or given as an input. Then, the model assigns about 5% and 95% of the remaining SOC to SOC1 and SOC2, respectively. The model performance when simulating SOC and nitrogen (N) dynamics strongly depends on the initialization process. The common methods (e.g. Basso et al., 2011) to initialize SOC pools deal mostly with carbon (C) mineralization processes and less with N. Dynamics of SOM, SOC, and soil organic N are linked in the CENTURY-DSSAT model through the C/N ratio of decomposing material that determines either mineralization or immobilization of N (Gijsman et al., 2002). The aim of this study was to evaluate an alternative method to initialize the SOC pools in the DSSAT-CENTURY model from apparent soil N mineralization (Napmin) field measurements by using automatic inverse calibration (simulated annealing). The results were compared with the ones obtained by the iterative initialization procedure developed by Basso et al., 2011.