Influence of sequential and mixed fermentations with non-Saccharomyces yeasts on the sensory profile of red wine

The aim of this work is to evaluate the influence of S. pombe and T. delbrueckii species on the sensory quality of red wine when used in sequential and mixed fermentations with S. cerevisiae.

Dietary xylanase increases hepatic vitamin E concentration of chickens fed wheat based diet

The study examined the effect of xylanase supplementation on apparent metabolizable energy (AME) and hepatic vitamin E and carotenoids in broiler chickens fed wheat based diets. A total of one hundred forty four male Ross 308 chickens were used in this study. Birds were randomly assigned to 3 dietary treatments (8 cages per treatment of 6 male broilers each) for 14 days from 7 to 21 day old. The control treatment was based on wheat-soyabean meal and was either unsupplemented or supplemented with either 1000 or 2000 xylanase units per kg diet. Orthogonal polynomial contrasts were used to test linear response to dietary xylanase activity. There was a positive linear relationship (P < 0.05) between dietary AME and doses of supplementary xylanase. A linear relationship (P < 0.05) was also observed between dosage of xylanase supplementation and hepatic vitamin E concentration and retention. In conclusion, xylanase supplementation improved dietary AME and increased hepatic vitamin E concentration which may have positive effects on the antioxidative status of the birds.

Partioning of dietary energy of chickens fed maize or wheat-based diets with and without a commercial blend of phytogenic feed additives

The aim of the study was to investigate the effects of a standardized mixture of a commercial blend of phytogenic feed additives containing 5% carvacrol, 3% cinnamaldehyde, and 2% capsicum on utilization of dietary energy and performance in broiler chickens. Four experimental diets were offered to the birds from 7 to 21 d of age. These included 2 basal control diets based on either wheat or maize that contained 215 g CP/kg and 12.13 MJ/kg ME and another 2 diets using the basal control diets supplemented with the plant extracts combination at 100 mg/kg diet. Each diet was fed to 16 individually penned birds following randomization. Dietary plant extracts improved feed intake and weight gain (P < 0.05) and slightly (P < 0.1) improved feed efficiency of birds fed the maize-based diet. Supplementary plant extracts did not change dietary ME (P > 0.05) but improved (P < 0.05) dietary NE by reducing the heat increment (P < 0.05) per kilogram feed intake. Feeding phytogenics improved (P < 0.05) total carcass energy retention and the efficiency of dietary ME for carcass energy retention. The number of interactions between type of diet and supplementary phytogenic feed additive suggest that the chemical composition and the energy to protein ratio of the diet may influence the efficiency of phytogenics when fed to chickens. The experiment showed that although supplementary phytogenic additives did not affect dietary ME, they caused a significant improvement in the utilization of dietary energy for carcass energy retention but this did not always relate to growth performance.

Advanced Characterisation of a Coffee Fermenting Tank by Multi-distributed Wireless Sensors: Spatial Interpolation and Phase Space Graphs

The fermentation stage is considered to be one of the critical steps in coffee processing due to its impact on the final quality of the product. The objective of this work is to characterise the temperature gradients in a fermentation tank by multi-distributed, low-cost and autonomous wireless sensors (23 semi-passive TurboTag® radio-frequency identifier (RFID) temperature loggers). Spatial interpolation in polar coordinates and an innovative methodology based on phase space diagrams are used. A real coffee fermentation process was supervised in the Cauca region (Colombia) with sensors submerged directly in the fermenting mass, leading to a 4.6 °C temperature range within the fermentation process. Spatial interpolation shows a maximum instant radial temperature gradient of 0.1 °C/cm from the centre to the perimeter of the tank and a vertical temperature gradient of 0.25 °C/cm for sensors with equal polar coordinates. The combination of spatial interpolation and phase space graphs consistently enables the identification of five local behaviours during fermentation (hot and cold spots).

Non-Destructive Global and Localized 2D T1/T2 NMR Relaxometry to Resolve Microstructure in Apples Affected by Watercore

Apples can be considered as having a complex system formed by several structures at different organization levels: macroscale (>100 μm) and microscale (<100 μm). This work implements 2D T1/T2 global and localized relaxometry sequences on whole apples to be able to perform an intensive non-destructive and non-invasive microstructure study. The 2D T1/T2 cross-correlation spectroscopy allows the extraction of quantitative information about the water compartmentation in different subcellular organelles. A clear difference is found as sound apples show neat peaks for water in different subcellular compartments, such as vacuolar, cytoplasmatic and extracellular water, while in watercore-affected tissues such compartments appear merged. Localized relaxometry allows for the predefinition of slices in order to understand the microstructure of a particular region of the fruit, providing information that cannot be derived from global 2D T1/T2 relaxometry.

Quantitative analysis of morphological changes in yeast colonies growing on solid medium: the eccentricity and Fourier indices

The colony shape of four yeast species growing on agar medium wasmeasured for 116 days by image analysis. Initially, all the colonies are circular, with regular edges. The loss of circularity can be quantitatively estimated by the eccentricity index, Ei, calculated as the ratio between their orthogonal vertical and horizontal diameters. Ei can increase from 1 (complete circularity) to a maximum of 1.17–1.30, depending on the species. One colony inhibits its neighbour only when it has reached a threshold area. Then, Ei of the inhibited colony increases proportionally to the area of the inhibitory colony. The initial distance between colonies affects those threshold values but not the proportionality, Ei/area; this inhibition affects the shape but not the total surface of the colony. The appearance of irregularities in the edges is associated, in all the species, not with age but with nutrient exhaustion. The edge irregularity can be quantified by the Fourier index, Fi, calculated by the minimum number of Fourier coefficients that are needed to describe the colony contour with 99% fitness. An ad hoc function has been developed in Matlab v. 7.0 to automate the computation of the Fourier coefficients. In young colonies, Fi has a value between 2 (circumference) and 3 (ellipse). These values are maintained in mature colonies of Debaryomyces, but can reach values up to 14 in Saccharomyces.All the species studied showed the inhibition of growth in facing colony edges, but only three species showed edge irregularities associated with substrate exhaustion. Copyright © 2014 John Wiley & Sons, Ltd.

A simple mathematical model that describes the growth of the area and the number of total and viable cells in yeast colonies

We propose a model, based on the Gompertz equation, to describe the growth of yeasts colonies on agar medium. This model presents several advantages: (i) one equation describes the colony growth, which previously needed two separate ones (linear increase of radius and of the squared radius); (ii) a similar equation can be applied to total and viable cells, colony area or colony radius, because the number of total cells in mature colonies is proportional to their area; and (iii) its parameters estimate the cell yield, the cell concentration that triggers growth limitation and the effect of this limitation on the specific growth rate. To elaborate the model, area, total and viable cells of 600 colonies of Saccharomyces cerevisiae, Debaryomyces fabryi, Zygosaccharomyces rouxii and Rhodotorula glutinis have been measured. With low inocula, viable cells showed an initial short exponential phase when colonies were not visible. This phase was shortened with higher inocula. In visible or mature colonies, cell growth displayed Gompertz-type kinetics. It was concluded that the cells growth in colonies is similar to liquid cultures only during the first hours, the rest of the time they grow, with near-zero specific growth rates, at least for 3 weeks.

Hyperspectral to multispectral imaging for detection of tree nuts and peanut traces in wheat flour.

In current industrial environments there is an increasing need for practical and inexpensive quality control systems to detect the foreign food materials in powder food processing lines. This demand is especially important for the detection of product adulteration with traces of highly allergenic products, such as peanuts and tree nuts. Manufacturing industries dealing with the processing of multiple powder food products present a substantial risk for the contamination of powder foods with traces of tree nuts and other adulterants, which might result in unintentional ingestion of nuts by the sensitised population. Hence, the need for an in-line system to detect nut traces at the early stages of food manufacturing is of crucial importance. In this present work, a feasibility study of a spectral index for revealing adulteration of tree nut and peanut traces in wheat flour samples with hyperspectral images is reported. The main nuts responsible for allergenic reactions considered in this work were peanut, hazelnut and walnut. Enhanced contrast between nuts and wheat flour was obtained after the application of the index. Furthermore, the segmentation of these images by selecting different thresholds for different nut and flour mixtures allowed the identification of nut traces in the samples. Pixels identified as nuts were counted and with the actual percentage of peanut adulteration. As a result, the multispectral system was able to detect and provide good visualisation of tree nut and peanut trace levels down to 0.01% by weight. In this context, multispectral imaging could operate in conjuction with chemical procedures, such as Real Time Polymerase Chain Reaction and Enzyme-Linked Immunosorbent Assay to save time, money and skilled labour on product quality control. This approach could enable not only a few selected samples to be assessed but also to extensively incorporate quality control surveyance on product processing lines.

VIS/NIR spectral signature for the identification of peanut contamination of powder foods

Visible-near infrared reflectance spectra are proposed for the characterization of IRMM 481 peanuts variety in comparison to powder food materials: wheat flour, milk and cocoa. Multidimensional analysis of reflectance spectra of powder samples shows a specific NIR band centred at 1200 nm that identifies peanut compared to the rest of food ingredients, regardless compaction level and temperature. Spectral range of 400-1000 nm is not robust for identification of blanched peanut. The visible range has shown to be reliable for the identification of pre-treatment and processing of unknown commercial peanut samples. A spectral index is proposed based on the combination of three wavelengths around 1200 nm that is 100% robust against pre-treatment (raw or blanched) and roasting (various temperatures and treatment duration).

Application of Independent Components Analysis with the JADE algorithm and NIR hyperspectral imaging for revealing food adulteration

In recent years, Independent Components Analysis (ICA) has proven itself to be a powerful signal-processing technique for solving the Blind-Source Separation (BSS) problems in different scientific domains. In the present work, an application of ICA for processing NIR hyperspectral images to detect traces of peanut in wheat flour is presented. Processing was performed without a priori knowledge of the chemical composition of the two food materials. The aim was to extract the source signals of the different chemical components from the initial data set and to use them in order to determine the distribution of peanut traces in the hyperspectral images. To determine the optimal number of independent component to be extracted, the Random ICA by blocks method was used. This method is based on the repeated calculation of several models using an increasing number of independent components after randomly segmenting the matrix data into two blocks and then calculating the correlations between the signals extracted from the two blocks. The extracted ICA signals were interpreted and their ability to classify peanut and wheat flour was studied. Finally, all the extracted ICs were used to construct a single synthetic signal that could be used directly with the hyperspectral images to enhance the contrast between the peanut and the wheat flours in a real multi-use industrial environment. Furthermore, feature extraction methods (connected components labelling algorithm followed by flood fill method to extract object contours) were applied in order to target the spatial location of the presence of peanut traces. A good visualization of the distributions of peanut traces was thus obtained

Combine use of Selected Schizosaccharomyces pombe and Lachancea thermotolerans Yeast Strains as an Alternative to the Traditional Malolactic Fermentation in Red Wine Production

Most red wines commercialized in the market use the malolactic fermentationprocess in order to ensure stability from a microbiological point of view. In this secondfermentation, malic acid is converted into L-lactic acid under controlled setups. Howeverthis process is not free from possible collateral effects that on some occasions produceoff-flavors, wine quality loss and human health problems. In warm viticulture regions suchas the south of Spain, the risk of suffering a deviation during the malolactic fermentationprocess increases due to the high must pH. This contributes to produce wines with highvolatile acidity and biogenic amine values. This manuscript develops a new red winemakingmethodology that consists of combining the use of two non-Saccharomyces yeast strains asan alternative to the traditional malolactic fermentation. In this method, malic acid is totallyconsumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilizationobjective, while Lachancea thermotolerans produces lactic acid in order not to reduce andeven increase the acidity of wines produced from low acidity musts. This technique reducesthe risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than thetraditional controls that have undergone the classical malolactic fermentation.

Multi-distributed wireless sensors for monitoring a long distance transport in a reefer container

The study of the temperature gradients in cold stores and containers is a critical issue in the food industry for the quality assurance of products during transport and for minimising losses. This work presents an analysis of the temperatures during the refrigerated transport of 4,320 kg of blueberries in a reefer (set point temperature at ?1ºC) on a container ship from Montevideo (Uruguay) to Verona (Italy). The monitoring was performed by using semi-passive RFID loggers (TurboTag cards). The objective was to carry out a multi-distributed supervision using low-cost, wireless and autonomous sensors for the characterisation of the distribution and spatial gradients of temperatures during a long distance transport. Data analysis shows spatial (phase space) and temporal sequencing diagrams and reveals a significant heterogeneity of temperature at different locations in the container, which highlights the ineffectiveness of a temperature control system based on a single sensor, as is usually done.

Supervisión multidistribuida para el control de procesos de conservación de productos agroalimentarios: optimización de calidad del producto

La presente Tesis está orientada al análisis de la supervisión multidistribuida de tres procesos agroalimentarios: el secado solar, el transporte refrigerado y la fermentación de café, a través de la información obtenida de diferentes dispositivos de adquisición de datos, que incorporan sensores, así como el desarrollo de metodologías de análisis de series temporales, modelos y herramientas de control de procesos para la ayuda a la toma de decisiones en las operaciones de estos entornos. En esta tesis se han utilizado: tarjetas RFID (TemTrip®) con sistema de comunicación por radiofrecuencia y sensor de temperatura; el registrador (i-Button®), con sensor integrado de temperatura y humedad relativa y un tercer prototipo empresarial, módulo de comunicación inalámbrico Nlaza, que integra un sensor de temperatura y humedad relativa Sensirion®. Estos dispositivos se han empleado en la conformación de redes multidistribuidas de sensores para la supervisión de: A) Transportes de producto hortofrutícola realizados en condiciones comerciales reales, que son: dos transportes terrestre de producto de IV gama desde Murcia a Madrid; transporte multimodal (barco-barco) de limones desde Montevideo (Uruguay) a Cartagena (España) y transporte multimodal (barco-camión) desde Montevideo (Uruguay) a Verona (Italia). B) dos fermentaciones de café realizadas en Popayán (Colombia) en un beneficiadero. Estas redes han permitido registrar la dinámica espacio-temporal de temperaturas y humedad relativa de los procesos estudiados. En estos procesos de transporte refrigerado y fermentación la aplicación de herramientas de visualización de datos y análisis de conglomerados, han permitido identificar grupos de sensores que presentan patrones análogos de sus series temporales, caracterizando así zonas con dinámicas similares y significativamente diferentes del resto y permitiendo definir redes de sensores de menor densidad cubriendo las diferentes zonas identificadas. Las metodologías de análisis complejo de las series espacio-temporales (modelos psicrométricos, espacio de fases bidimensional e interpolaciones espaciales) permitieron la cuantificación de la variabilidad del proceso supervisado tanto desde el punto de vista dinámico como espacial así como la identificación de eventos. Constituyendo así herramientas adicionales de ayuda a la toma de decisiones en el control de los procesos. Siendo especialmente novedosa la aplicación de la representación bidimensional de los espacios de fases en el estudio de las series espacio-temporales de variables ambientales en aplicaciones agroalimentarias, aproximación que no se había realizado hasta el momento. En esta tesis también se ha querido mostrar el potencial de un sistema de control basado en el conocimiento experto como es el sistema de lógica difusa. Se han desarrollado en primer lugar, los modelos de estimación del contenido en humedad y las reglas semánticas que dirigen el proceso de control, el mejor modelo se ha seleccionado mediante un ensayo de secado realizado sobre bolas de hidrogel como modelo alimentario y finalmente el modelo se ha validado mediante un ensayo en el que se deshidrataban láminas de zanahoria. Los resultados sugirieron que el sistema de control desarrollado, es capaz de hacer frente a dificultades como las variaciones de temperatura día y noche, consiguiendo un producto con buenas características de calidad comparables a las conseguidas sin aplicar ningún control sobre la operación y disminuyendo así el consumo energético en un 98% con respecto al mismo proceso sin control. La instrumentación y las metodologías de análisis de datos implementadas en esta Tesis se han mostrado suficientemente versátiles y transversales para ser aplicadas a diversos procesos agroalimentarios en los que la temperatura y la humedad relativa sean criterios de control en dichos procesos, teniendo una aplicabilidad directa en el sector industrial ABSTRACT This thesis is focused on the analysis of multi-distributed supervision of three agri-food processes: solar drying, refrigerated transport and coffee fermentation, through the information obtained from different data acquisition devices with incorporated sensors, as well as the development of methodologies for analyzing temporary series, models and tools to control processes in order to help in the decision making in the operations within these environments. For this thesis the following has been used: RFID tags (TemTrip®) with a Radiofrequency ID communication system and a temperature sensor; the recorder (i-Button®), with an integrated temperature and relative humidity and a third corporate prototype, a wireless communication module Nlaza, which has an integrated temperature and relative humidity sensor, Sensirion®. These devices have been used in creating three multi-distributed networks of sensors for monitoring: A) Transport of fruits and vegetables made in real commercial conditions, which are: two land trips of IV range products from Murcia to Madrid; multimodal transport (ship - ship) of lemons from Montevideo (Uruguay) to Cartagena (Spain) and multimodal transport (ship - truck) from Montevideo (Uruguay) to Verona (Italy). B) Two coffee fermentations made in Popayan (Colombia) in a coffee processing plant. These networks have allowed recording the time space dynamics of temperatures and relative humidity of the processed under study. Within these refrigerated transport and fermentation processes, the application of data display and cluster analysis tools have allowed identifying sensor groups showing analogical patterns of their temporary series; thus, featuring areas with similar and significantly different dynamics from the others and enabling the definition of lower density sensor networks covering the different identified areas. The complex analysis methodologies of the time space series (psychrometric models, bi-dimensional phase space and spatial interpolation) allowed quantifying the process variability of the supervised process both from the dynamic and spatial points of view; as well as the identification of events. Thus, building additional tools to aid decision-making on process control brought the innovative application of the bi-dimensional representation of phase spaces in the study of time-space series of environmental variables in agri-food applications, an approach that had not been taken before. This thesis also wanted to show the potential of a control system based on specialized knowledge such as the fuzzy logic system. Firstly, moisture content estimation models and semantic rules directing the control process have been developed, the best model has been selected by an drying assay performed on hydrogel beads as food model; and finally the model has been validated through an assay in which carrot sheets were dehydrated. The results suggested that the control system developed is able to cope with difficulties such as changes in temperature daytime and nighttime, getting a product with good quality features comparable to those features achieved without applying any control over the operation and thus decreasing consumption energy by 98% compared to the same uncontrolled process. Instrumentation and data analysis methodologies implemented in this thesis have proved sufficiently versatile and cross-cutting to apply to several agri-food processes in which the temperature and relative humidity are the control criteria in those processes, having a direct effect on the industry sector.

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.

Influence of sequential fermentations with Torulaspora delbrueckii and Saccharomyces cerevisiae on wine quality

Torulaspora delbrueckii is a non-Saccharomyces yeast with interesting metabolic and physiological properties of potential use in oenology. This work examines the fermentative behaviour of five strains of T. delbrueckii in sequential fermentations with Saccharomyces cerevisiae, analysing the formation of aromatic compounds, polyalcohols and pigments. The fermentative power of these five strains ranged between 7.6 and 9.0% v/v ethanol; the associated volatile acidity was 0.2e0.7 g/l acetic acid. The production of glycerol was inferior to that of S. cerevisiae alone. The mean 2,3-butanediol concentration reached in single-culture S. cerevisiae fermentations was 73% higher than in the five sequential T. delbrueckii/S. cerevisiae fermentations. However, these fermentations produced larger quantities of diacetyl, ethyl lactate and 2-phenylethyl acetate than single-culture S. cerevisiae fermentation. 3-ethoxy propanol was produced only in the sequential fermentations. The five sequential fermentations produced smaller quantities of vitisin A and B than single-culture S. cerevisiae fermentation. In tests performed prior to the addition of the S. cerevisiae in the sequential fermentations, none of the T. delbrueckii strains showed any extracellular hydroxycinnamate decarboxylase activity. They therefore produced no vinyl phenolic pyranoanthocyanins.