869 resultados para Longoni, Ana


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The flow and vaporization behaviors of long-chain esters of varying molecular weights (300-900) ana branching (linear, Y-shaped, and +-shaped molecules) have been studied. The flow behavior is found to depend on the structure as well as the molecular weight. Below a molecular weight of 600, the molecules flow wholly but above this, segmental motion occurs, and the flow becomes independent of the molecular weight which is explained from the blob model. The blob concept demonstrates that the hole of a size of about 11 angstrom is needed for the flow to occur and it is much less than the size of the molecule. The blob size is observed to slightly decrease along the series linear and Y- and +-branched esters. The heat of vaporization is found to be independent of the molecular structure since the molecules acquire a coiled spherical shape during vaporization and hence depends only on the molecular weight. A significant structural effect is observed for the esters on their glass transition temperature (T(g)). The T(g) vs molecular weight plot displays contrasting trend for linear and +-branched esters, with Y esters showing an intermediate behavior. It is explained from their molecular packing and entanglement as visualized by the blob model.

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The most important objective of the present study was to explain why cationic lipid (CL)-mediated delivery of plasmid DNA (pDNA) is better than that of linear DNA in gene therapy, a question that, until now, has remained unanswered. Herein for the first time we experimentally show that for different types of CLs, pDNA, in contrast to linear DNA, is compacted with a large amount of its counterions, yielding a lower effective negative charge. This feature has been confirmed through a number of physicochemical and biochemical investigations. This is significant for both in vitro and in vivo transfection studies. For an effective DNA transfection, the lower the amount of the CL, the lower is the cytotoxicity. The study also points out that it is absolutely necessary to consider both effective charge ratios between CL and pDNA and effective pDNA charges, which can be determined from physicochemical experiments.

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Lipoplexes formed by the pEGFP-C3 plasmid DNA (pDNA) and lipid mixtures containing cationic gemini surfactant of the 1,2-bis(hexadecyl dimethyl ammonium) Acmes family referred to as C16CnC16, where n = 2 3, 5, or 12, and the zwitterionic helper lipid, 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) have been studied from a wide variety of physical, chemical, and biological standpoints. The study has been carried out using several experimental methods, such as zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), cryo-TEM, gene transfection, cell viability/cytotoxicity, and confocal fluorescence microscopy. As reported recently in a communication (J. Am. Chem. Soc. 2011, 133, 18014), the detailed physicochemical and biological studies confirm that, in the presence of the studied series lipid mixtures, plasmid DNA is compacted with a large number of its associated Na+ counterions. This in turn yields a much lower effective negative charge, q(pDNA)(-), a value that has been experimentally obtained for each mixed lipid mixture. Consequently, the cationic lipid (CL) complexes prepared with pDNA and CL/DOPE mixtures to be used in gene transfection require significantly less amount of CL than the one estimated assuming a value of q(DNA)(-) = -2. This drives to a considerably lower cytotoxicity of the gene vector. Depending on the CL molar composition, alpha, of the lipid mixture, and the effective charge ratio of the lipoplex, rho(eff), the reported SAXS data indicate the presence of two or three structures in the same lipoplex, one in the DOPE-rich region, other in the CL-rich region, and another one present at any CL composition. Cryo-TEM and SAXS studies with C16CnC16/DOPE-pDNA lipoplexes indicate that pDNA is localized between the mixed lipid bilayers of lamellar structures within a monolayer of similar to 2 nm. This is consistent with a highly compacted supercoiled pDNA conformation compared with that of linear DNA. Transfection studies were carried out with HEK293T, HeLa, CHO, U343, and H460 cells. The alpha and rho(eff) values for each lipid mixture were optimized on HEK293T cells for transfection, and using these values, the remaining cells were also transfected in absence (-FBS-FBS) and presence (-FBS+FBS) of serum. The transfection efficiency was higher with the CLs of shorter gemini spacers (n = 2 or 3). Each formulation expressed GFP on pDNA transfection and confocal fluorescence microscopy corroborated the results. C16C2C16/DOPE mixtures were the most efficient toward transfection among all the lipid mixtures and, in presence of serum, even better than the Lipofectamine2000, a commercial transfecting agent Each lipid combination was safe and did not show any significant levels of toxicity. Probably, the presence of two coexisting lamellar structures in lipoplexes synergizes the transfection efficiency of the lipid mixtures which are plentiful in the lipoplexes formed by CLs with short spacer (n = 2, 3) than those with the long spacer (n = 5, 12).

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Lipoplex-type nanoaggregates prepared from pEGFP-C3 plasmid DNA (pDNA) and mixed liposomes, with a gemini cationic lipid (CL) 1,2-bis(hexadecyl imidazolium) alkanes], referred as (C(16)Im)(2)C-n (where C-n is the alkane spacer length, n = 2, 3, 5, or 12, between the imidazolium heads) and DOPE zwitterionic lipid, have been analyzed by zeta potential, gel electrophoresis, SAXS, cryo-TEM, fluorescence anisotropy, transfection efficiency, fluorescence confocal microscopy, and cell viability/cytotoxicity experiments to establish a structure-biological activity relationship. The study, carried out at several mixed liposome compositions, alpha, and effective charge ratios, rho(eff), of the lipoplex, demonstrates that the transfection of pDNA using CLs initially requires the determination of the effective charge of both. The electrochemical study confirms that CLs with a delocalizable positive charge in their headgroups yield an effective positive charge that is 90% of their expected nominal one, while pDNA is compacted yielding an effective negative charge which is only 10-25% than that of the linear DNA. SAXS diffractograms show that lipoplexes formed by CLs with shorter spacer (n = 2, 3, or 5) present three lamellar structures, two of them in coexistence, while those formed by CL with longest spacer (n = 12) present two additional inverted hexagonal structures. Cryo-TEM micrographs show nanoaggregates with two multilamellar structures, a cluster-type (at low alpha value) and a fingerprint-type, that coexist with the cluster-type at moderate alpha composition. The optimized transfection efficiency (TE) of pDNA, in HEK293T, HeLa, and H1299 cells was higher using lipoplexes containing gemini CLs with shorter spacers at low a value. Each lipid formulation did not show any significant levels of toxicity, the reported lipoplexes being adequate DNA vectors for gene therapy and considerably better than both Lipofectamine 2000 and CLs of the 1,2-bis(hexadecyl ammnoniun) alkane series, recently reported.

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Despite intense research efforts that have provided enormous insight, cancer continues to be a poorly understood disease. There has been much debate over whether the cancerous state can be said to originate in a single cell or whether it is a reflection of aberrant behaviour on the part of a `society of cells'. This article presents, in the form of a debate conducted among the authors, three views of how the problem might be addressed. We do not claim that the views exhaust all possibilities. These views are (a) the tissue organization field theory (TUFT) that is based on a breakdown of tissue organization involving many cells from different embryological layers, (b) the cancer stem cell (CSC) hypothesis that focuses on genetic and epigenetic changes that take place within single cells, and (c) the proposition that rewiring of the cell's protein interaction networks mediated by intrinsically disordered proteins (IDPs) drives the tumorigenic process. The views are based on different philosophical approaches. In detail, they differ on some points and agree on others. It is left to the reader to decide whether one approach to understanding cancer appears more promising than the other.

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Lipoplex nano-aggregates have been analyzed through biophysical characterization (electrostatics, structure, size and morphology), and biological studies (transfection efficiency and cell viability) in five cancer cell lines. Lipoplexes were prepared from pEGFP-C3 plasmid DNA (pDNA) and mixed liposomes, constituted by a zwitterionic lipid (DOPE) and a gemini cationic lipid (GCL) synthesized in this work, bis(hexadecyl dimethyl ammonium) oxyethylene], referred to as (C16Am)(2)(C2O)(n), (where n is the oxyethylene spacer length, n = 1, 2 or 3, between the ammonium heads). Cryo-TEM micrographs show nano-aggregates with two multilamellar structures, a cluster-type (at low-to-medium GCL composition) and a fingerprint-type that coexists with the cluster-type at medium GCL composition and appears alone at high GCL composition. SAXS diffractograms show that these lipoplexes present three lamellar structures, two of them coexisting at low and high GCL composition. The optimized transfection efficiency (TE) of pDNA was higher for lipoplexes containing GCLs with a longer (n = 3) or shorter (n = 1) polyoxyethylene spacer, at high GCL composition (alpha - 0.7) with low charge ratio (rho(eff) 2). In the all cancer cell lines studied, the TE of the optimized formulations was much better than those of both lipofectamine 2000 and lipoplexes with GCLs of the bis(hexadecyl dimethyl ammonium) alkane series recently reported. Probably, (a) the coexistence of two lamellar structures at high GCL composition synergizes the TE of these lipid vectors, (b) the orientation of the polyoxyethylene region in (C16Am)(2)(C2O)(3)/DOPE may occur in such a way that the spacing between two cationic heads becomes smaller than that in (C16Am)(2)(C2O)(2)/DOPE which is poor in terms of TE, and (c) the synergistic interactions between serum proteins and (C16Am)(2)(C2O)(n)/DOPE-pDNA lipoplexes containing a polyoxyethylene spacer improve TE, especially at high GCL content. Lipoplexes studied here show very low levels of toxicity, which confirm them as improved vectors of pDNA in gene therapy.

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Lipoplex nano-aggregates constituted of plasmid DNA (pDNA) pEGFP-C3 and mixed cationic liposomes, consisting of several percentages of a gemini cationic lipid (GCL) of the 1,2-bis(hexadecyl imidazolium) oxyethylene series, referred to as (C(16)Im)(2)(C2O)(n), with oxyethylene spacers (n = 1, 2 or 3) between the imidazolium cationic groups and the DOPE zwitterionic helper lipid, have been characterized by various biophysical and biological approaches carried out at several GCL compositions (alpha), and either the mass or the effective charge ratio of the lipoplex. The electrochemical study by zeta-potential confirms that the three GCLs yield a 10% lower effective charge than the nominal one, while compacted pDNA yields only a 25% effective negative charge. The SAXS study reveals, irrespective of the spacer length (n) and effective charge ratio (rho(eff)), the presence of two lamellar structures, i.e., one (L-alpha,L-main) in the whole GCL composition and another (L-alpha,L-DOPE,L-rich) with higher periodicity values that coexists with the previous one at low GCL composition (alpha = 0.2). The cryo-TEM analysis shows two types of multilamellar structures consisting of cationic lipidic bilayers with pDNA sandwiched between them: a cluster-type (C-type) at low alpha = 0.2 and a fingerprint-type (FP-type) at alpha >= 0.5, both with similar interlamellar spacing (d) in agreement with the L-alpha,L-main structure determined by SAXS. Transfection efficacies (TEs) of each lipid mixture were determined in four different cell lines (HEK293T, HeLa, Caco-2 and A549) at several alpha and rho(eff) values in the absence and presence of serum (FBS). The optimized formulations (alpha = 0.2 and rho(eff) = 2.0) substantially transfect cells much better than a commercial transfection reagent, Lipofectamine 2000 and previously studied efficient lipoplexes containing other cationic head groups or spacers both in the absence and presence of serum. The activity of optimized formulations may be attributed to the combination of several factors, such as: (a) the fusogenic character of DOPE which results in higher fluidity of the lipoplexes at alpha = 0.2, (b) the coexistence of two lamellar structures at alpha = 0.2 that synergizes the TE of these lipid vectors, and mainly (c) the higher biocompatibility of the GCLs reported in this work due to the presence of two imidazolium cationic groups together with an oligo-oxyethylene spacer. The length of the spacer in the GCL seems to have less impact, although (C(16)Im)(2)(C2O)(n)/DOPEpDNA lipoplexes with n = 1 and 3 show higher gene transfection than n = 2. All the optimum formulations reported herein are all highly efficient with negligible levels of toxicity, and thus, may be considered as very promising gene vectors for in vivo applications.

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Determining the concentrations of acetylcholine (ACh) and choline (Ch) is clinically important. ACh is a neurotransmitter that acts as a key link in the communication between neurons in the spinal cord and in nerve skeletal junctions in vertebrates, and plays an important role in transmitting signals in the brain. A bienzymatic sensor for the detection of ACh was prepared by co-immobilizing choline oxidase (ChO) and acetylcholinesterase (AChE) on graphene matrix/platinum nanoparticles, and then electrodepositing them on an ITO-coated glass plate. Graphene nanoparticles were decorated with platinum nanoparticles and were electrodeposited on a modified ITO-coated glass plate to form a modified electrode. The modified electrode was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) studies. The optimum response of the enzyme electrode was obtained at pH 7.0 and 35 degrees C. The response time of this ACh-sensing system was shown to be 4 s. The linear range of responses to ACh was 0.005-700 mu M. This biosensor exhibits excellent anti-interferential abilities and good stability, retaining 50% of its original current even after 4 months. It has been applied for the detection of ACh levels in human serum samples.

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Plasmon rulers, consisting of pairs of gold nanoparticles, allow single-molecule analysis without photobleaching or blinking; however, current plasmon rulers are irreversible, restricting detection to only single events. Here, we present a reversible plasmon ruler, comprised of coupled gold nanoparticles linked by a single aptamer, capable of binding individual secreted molecules with high specificity. We show that the binding of target secreted molecules to the reversible plasmon ruler is characterized by single-molecule sensitivity, high specificity, and reversibility. Such reversible plasmon rulers should enable dynamic and adaptive live-cell measurement of secreted single molecules in their local microenvironment.

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El presente trabajo se realizó en el Arboretum de la Universidad Nacional Agraria. El objetivo del estudio es identificar las características morfológicas de cada cultivar clónal como: largo de la hoja, número y largo de folíolo, ancho de la base y ápice, sobrevivencia, número y tamaño de rebrotes. La metodología consta de tres etapas: recorrido entre los municipios de Masaya y Granada para la obtención de el material vegetativo con una dimensión de 1.30 m de altura y 10cm de diámetro por estaca. Establecimiento de plantación de 10 cultivares de Spondia sp. para un total de 40 estacas con una distancia de 3 x 3 m. para un área total de 243m2. Toma de datos cada seis meses para realizar el análisis estadístico empleando el programa INFOSTAT y mediante el encadenamiento simple (Distancia Euclidiana) se dividieron los 9 cultivares en 4 grupos y se procedió a realizar un ANOVA obteniendo que los cultivares clónales presentan diferencia significativa (p<000.1) en cuanto a sus variables. Como resultados se obtiene una sobrevivencia del 100% en los cultivares clónales de jocote perro, jocote agosteño, jocote rosa y jocote diente de perro, siendo el jocote de cocer sabanero el que no sobrevivió. Dentro de las características morfológicas sobresaliente se identifica como el raquis más largo al jocote rosa, con un promedio de 23.4 cm y el de menor longitud con 13.8 es el jocote agosteño, el mayor promedio de folíolos por hoja lo tiene el jocote bejuco con 20 y el de menor promedio el jocote agosteño con 14. Entre las variables número y largo de foliolo, largo del raquis, ancho de la base y ápice mediante un ANOVA se obtuvo diferencia significativa (p<000.1). De acuerdo a los estudios morfológicos aplicados a los nueve cultivares clónales sobrevivientes se ubican dentro de la especie Spondias purpurea.

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Se realizó un experimento con 30 cerdos comerciales en las etapas de desarrollo y engorde, con una duración de 120 días (60 días por etapa) con el objetivo de evaluar el efecto de la inclusión de yuca y suero en la alimentación. Sobre consumo, ganancia medía diaria (GMD), conversión alimenticia y utilidad económica. Los cerdos fueron asignados aleatoriamente en 3 grupos de 10 cerdos cada uno, de pesos similares con 60 días de edad, utilizándose un Diseño Completo al Azar (D.C.A. ). Se encontró diferencias significativas (P<0.05) en las categorías de desarrollo y engorde en cuanto a ganancia medía diaria del tratamiento T 1 respecto a los tratamientos T2 y T3. Las raciones experimentales estaban constituidas por: T1 (testigo) concentrado comercial, T2 base (50% maíz., 50% semolina y sal) y suero y el T3 la misma base, suero y yuca. En la etapa de desarrollo los consumos de alimento, ganancia media diaria y conversión alimenticia promedio por cerdo fueron de: 63.48, 189.57 y 218.45 kg; 384.33, 174.33 y 158.33 gramos/día; 2.75, 19.95 y 20.88 para T1, T2 y T3 respectivamente. En la etapa de engorde, los consumos de alimento, ganancia media diaria y conversión alimenticia promedia por cerdo fueron de: 164.98, 343.44 y 355.87 kg; 822.03, 398.65 y 328.80 gramos/día; 3.34, 14.36 y 18.03. El análisis económico evidencia que las mejores utilidades se obtuvieron con el T2 en la etapa de engorde con C$90.61/cerdo. Se concluye que se puede suministrar raciones para la alimentación de cerdos en engorde con subproductos lácteos y agrícolas con el fin de disminuir los costos por alimento.

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En el presente estudio se evaluaron tres periodos de alimento de retiro o acabado en broilers (o, 3, 6 y 9 días antes de la matanza) el cual contiene 3,225 kcal EM/kg de alimento y 19% de proteína, ajustándose a los requerimientos nutricionales recomendados por Arbor Acres Farm Inc., (1992), en especial a la proporción energia-proteina. La evaluación tuvo lugar en la Granja Buenos Aires propiedad de la Empresa Tip-Top Industrial, S.A., con una duración de 42 días, en donde se utilizaron 800 pollos de engorde sin sexar (mixtos) de la línea Peterson-Arbor Acres de un día de edad, dichos pollos fueron distribuidos aleatoriamente en cuatro tratamientos: T1 suministro de alimento de retiro por nueve días), T2 suministro de alimento de retiro por seis días), T3, (Suministro de alimento de retiro por tres días) y T4 (testigo, con cero días de alimento de retiro), con cuatro repeticiones cada uno, distribuidos en un Diseño completamente al Azar sometidos a la prueba de rangos múltiples de Duncan. Las variables estudiadas fueron: consumo de alimento, peso vivo, ganancia de peso, conversión alimenticia, mortalidad vs viabilidad, rendimiento en la canal, análisis económico. No se encontraron diferencias significativas (P<0.05) entre los tratamientos al final del ensayo para el consumo de alimento, conversión alimenticia y costo de alimento, pero no así ara peso vivo y ganancia de peso. El porcentaje de mortalidad acumulada total fue de 2.5% y los rendimientos de la canal fueron: (90.74)T1 (88.98)T2, (85.86)T3,y (90.31)T4, Se corolo que el periodo de suministro de alimento de retiro que permitió los mejores pesos vivos, ganancias de pesos y conversiones alimenticias fue el periodo de tres días (T3), sin embargo, obtuvo el mayor consumo total para generar un peso promedio en la canal de 3.34 lbs, además, presentó el mayor costo alimenticio entre los tratamientos experimentales (T1, T2, T3) y bajo rendimiento en la canal, contrario al T1 que aunque los parámetros productivos fueron menores que el T3 obtuvo el mayor rendimiento en la canal con un menor costo alimenticio. El alimento de retiro no ejerció efecto sobre las variables estudiadas lo que demuestra que dicho alimento pueda ser una alternativa viable para disminuir costo alimenticio.

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Nicaragua produce 147,000 lb/mes de subproducto de la pesca del camarón y del pescado los cuales son destinados a la basura. Con e l objetivo de proporcionar un método de conservación que conlleve a utilizar estos desperdicios en la alimentación animal se realizó el presente estudio que consistió camarón, en elaborar tratados microensilajes con subproducto con ácido sulfúrico (5%, del 10%) volumen/peso utilizando de 2 Kg cada uno. 20 microsilos de PVC con capacidad se dividieron en 2 tratamientos evaluándolos a diferentes periodos de tiempo (7, 15, 30, 45 y 60 días), realizándose un DCA con arreglo unifactorial, análisis bromatológicos, determinación de AGV y pH. Todos los ensilados presentaron predominancia de la fermentación láctica, los contenidos de proteína bruta variaron en un rango de 40.70 a 50.20% según los tratamientos. Todos los ensilajes se pueden utilizar a los 7 días después de ensilarse. El tratamiento de menor costo resulto ser el T1 (5% H2so4 ) con 0.56 USA/Kg.

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En el presente estudio se compararon cuatro dietas con diferentes niveles de energía 2,940, 3,040, 3,140 y 3,240 (Kcal EM/Kg de alimento), los cuales fueron ajustados a los requerimientos nutricionales en cuanto a la relación caloría proteína recomendados por la Hubbard Farm, Inc. (1991). Durante un periodo de 56 días los broilers fueron evaluados en la granja avícola "Buenos Aires" que pertenece a la Tip Top Industrial S.A. se utilizaron 803 pollos de engorde de un día de nacidos de la línea Hubbard, los que fueron distribuidos aleatoriamente en tres unidades experimentales proporcionales a los cuatro tratamientos existentes, estos distribuidos en un diseño completamente al azar y sometidos a la prueba de Tukey el que proporciona la superioridad existente entre tratamientos. Se encontraron diferencias significativas (P<0.05) entre tratamientos en las variables peso vivo, ganancia de peso y conversión alimenticia pero no así para el consumo alimenticio. Los tratamientos T1 T2, T3 y T4 presentaron mortalidad al final del periodo de 2.57, 4.41, 7.88 y 10.89 (%) respectivamente y rendimiento en la canal del 84.45 % para todos los tratamientos. Se concluyó que las dietas más energéticas proporcionaban las mejores conversiones alimenticias y las mayores ganancias de peso. Y a medida que se incrementaba el nivel energético en las dietas los costos alimenticios también se incrementaban. La fórmula más económica es el T1 (2940 Kcal EM/Kg de alimento, 21.63 y 18.89 % de proteína para inicio y finalización) durante todas las semanas de estudio. El momento óptimo para la matanza es la sexta semana para el T 1 (2940 Kcal EM/Kg de alimento, 21.63 y 18.89 % de proteínas) y T2 (3040 Kcal EM/Kg de alimento, 22.36 y 19.15 % de proteínas) por efecto de los costos alimenticios.

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El presente estudio se realizó en la finca “Santa Rosa” propiedad de la Universidad Nacional Agraria, localizada geográficamente a los 12°08’15’’ latitud Norte y a los 86°09’36’’ longitud Este, en el departamento de Managua, con el objetivo de evaluar el comportamiento productivo de ovinos alimentados con una dieta basal de pasto guinea (Panicum máximum Jacq) y suplementados con diferentes niveles de Moringa oleífera. Se utilizaron 18 corderos mestizos (Pelibuey x Blackbelly) con pesos iníciales promedio de 20 ± 2 kg, los cuales fueron desparasitados, vitaminados y distribuidos en un Diseño Completamente Aleatorio con tres tratamientos: TI Panicum máximumad-libitum, TII P. máximum ad-libitum + 0.35 kg MSM. oleífera, TIII P. máximum ad-libitum + 0.50 kg MSM. oleífera. Las variables estudiadas fueron: consumo diario de MS, ganancia media diaria y conversión alimenticia. Se realizó análisis de varianza y comparaciones de medias con la Prueba de Tukey utilizando MINITAB, versión 12.0. Los resultados de los análisis de varianza (P<0,05) mostraron que el m ayor consumo total de MS, ganancia media diaria y conversión alimenticia se obtiene con el TIII (0.8 kg MS/animal/día, 117.97 g/animal/día y 6.78) el que difiere e stadísticamente (P< 0.01) del TI (0.57 kg MS/animal/día, 30.85 g/animal/día y 18.47) pero no difiere significativamente (P> 0.05)del TII (0.73 kg MS/animal/día, 90.91 g/animal/día y 8.02).En conclusión el forraje de M. oleífera como suplemento proteico para ovinos consumiendo una dieta basal de P.máximum incrementa la ganancia de peso y mejora el consumo total de MS y la conversión alimenticia.