Effects of the correction of particle microbial contamination and particle transit model in the rumen on in situ protein evaluation of grass hays.

Effects of considering the particle comminution rate -kc- in addition to particle rumen outflow -kp- and the ruminal microbial contamination on estimates of by-pass and intestinal digestibility of DM, organic matter and crude protein were examined in perennial ryegrass and oat hays. By-pass kc-kp-based values of amino acids were also determined. This study was performed using particle transit, in situ and 15N techniques on three rumen and duodenum-cannulated wethers. The above estimates were determined using composite samples from rumen-incubated residues representative of feed by-pass. Considering the comminution rate, kc, modified the contribution of the incubated residues to these samples in both hays and revealed a higher microbial contamination, consistently in oat hay and only as a tendency for crude protein in ryegrass hay. Not considering kc or rumen microbial contamination overvalued by-pass and intestinal digestibility in both hays. Therefore, non-microbial-corrected kp-based values of intestinal digested crude protein were overestimated as compared with corrected and kc-kp-based values in ryegrass hay -17.4 vs 4.40%- and in oat hay -5.73 vs 0.19%-. Both factors should be considered to obtain accurate in situ estimates in grasses, as the protein value of grasses is very conditioned by the microbial synthesis derived from their ruminal fermentation. Consistent overvaluations of amino acid by-pass due to not correcting microbial contamination were detected in both hays, with large variable errors among amino acids. A similar degradation pattern of amino acids was recorded in both hays. Cysteine, methionine, leucine and valine were the most degradation-resistant amino acids.

Changes in rumen microbes in sheep in response to dietary forage quality and relationships with fermentation characteristics

An in vitro experiment was carried out using the Hohenheim gas production technique to evaluate 24-h gas production, apparently and truly degraded dry matter (DM), partitioning factor (PF), short chain fatty acids, crude protein (CP) and carbohydrate (CHO) fractionation of grass and multipurpose tree species (MPTS) foliage diets. Four grasses and three MPTS were used to formulate 12 diets of equal mixtures (0.5:0.5 on DM basis) of each grass with each MPTS. In vitro gas production was terminated after 24 h for each diet. True DM degradability was measured from incubated samples and combined with gas volume to estimate PF. Diets had greater (P<0.001) CP (102–183 g/kg DM) content than sole grasses (66–131 g/kg DM) and lower (P<0.001) concentrations of fibre fractions. Contrary to in vitro apparently degraded DM, in vitro truly degraded DM coefficient was greater (P<0.001) in diets (0.63–0.77) than in sole grasses (0.48–0.68). The PF was on average higher in diets than in sole grasses. The proportion of potentially degradable CP fractions (A1, B1, B2 and B3, based on the Cornell Net Carbohydrate and Protein System) in the diets ranged from 971 to 989 g/kg CP. Crude protein fractions, A and B2 were greater in diets but B1 and B3 fractions were less in diets than in sole grasses. A similar trend was also observed in the CHO fractions. Results showed that the nutritive value of the four grasses was improved when MPTS leaves were incorporated into the diet and this could ensure higher productivity of the animals.

Caracterización y optimización de estructuras biofotónicas y técnicas avanzadas de interrogación para desarrollo de biosensores ópticos

El núcleo fundamental de esta tesis doctoral es un modelo teórico de la interacción de la luz con un tipo particular de biosensor óptico. Este biosensor se compone de dos regiones: en la región inferior puede haber capas de materiales con diferentes espesores y propiedades ópticas, apiladas horizontalmente; en la zona superior, sobre la que incide directamente el haz de luz, puede haber estructuras que hacen que las propiedades ópticas cambien tanto en el plano horizontal como en la dirección vertical. Estos biosensores responden ópticamente de forma diferente al ser iluminados dependiendo de que su superficie externa esté, en mayor o menor medida, recubierta con diferentes tipos de material biológico. En esta tesis se define un modelo analítico aproximado que permite simular la respuesta óptica de biosensores con estructuras en su región más externa. Una vez comprobada la validez práctica del modelo mediante comparación con medidas experimentales, éste se utiliza en el diseño de biosensores de rendimiento óptimo y en la definición de nuevas técnicas de interrogación óptica. En particular, el sistema de transducción IROP (Increased Relative Optical Power), basado en el efecto que produce la presencia de material biológico, en la potencia total reflejada por la celda biosensora en determinados intervalos espectrales, es uno de los sistemas que ha sido patentado y es objeto de desarrollo por la empresa de base tecnológica BIOD [www.biod.es/], estando ya disponibles en este momento varios dispositivos de diagnóstico basados en esta idea. Los dispositivos basados en este sistema de transducción han demostrado su eficiencia en la detección de proteínas y agentes infecciosos como los rotavirus y el virus del dengue. Finalmente, el modelo teórico desarrollado se utiliza para caracterizar las propiedades ópticas de algunos de los materiales de los que se fabrican los biosensores, así como las de las capas de material biológico formadas en las diferentes fases de un inmunoensayo. Los parámetros ópticos de las capas mencionadas se obtienen mediante el método general de ajuste por mínimos cuadrados a las curvas experimentales obtenidas en los inmunoensayos. ABSTRACT The core of this thesis is the theoretical modeling of the interaction of light with a particular type of optical biosensor. This biosensor consists of two parts: in the lower region may have layers of materials with different thicknesses and optical properties, stacked horizontally; at the top, on which directly affects the light beam, there may be structures that make optical properties change in both, the horizontal and in the vertical direction. These biosensors optically respond differently when illuminated depending on its external surface is greater or lesser extent, coated with different types of biological material. In this thesis an approximate analytical model to simulate the optical response of biosensors with structures in its outer region is defined. After verifying the practical validity of the model by comparison with experimental measurements, it is used in the design of biosensors with optimal performance and the definition of new optical interrogation techniques. In particular, the transduction system IROP (Increased Relative Optical Power) based on the effect of the presence of biological material in the total power reflected from the biosensor cell in certain spectral ranges, has been patented and is under development by the startup company BIOD [www.biod.es/], being already available at this time, several diagnostic devices based on this idea. Devices based on this transduction system have proven their efficiency in detecting proteins and infectious agents such as rotavirus and virus of dengue. Finally, the developed theoretical model is used to characterize the optical properties of some of the materials from which biosensors are fabricated, as well as the optical properties of the biological material layers formed at different stages of an immunoassay. The optical parameters of the layers above are obtained by the general method of least squares fit to the experimental curves obtained in immunoassays.