In vitro effect of deoxynivalenol (DON) mycotoxin on porcine reproductive and respiratory syndrome virus replication.

Deoxynivalenol (DON) is a mycotoxin produced by Fusarium spp. Among monogastric farm animals, swine are the most susceptible to DON as it markedly reduces feed intake and decreases weight gain. DON has also been shown to increase susceptibility to viral infections; therefore the objective of this study was to investigate in vitro impact of DON on porcine reproductive and respiratory syndrome virus (PRRSV). Permissive cells were infected or not with PRRSV and were treated with increasing concentrations of DON. Cell survival and mortality were evaluated by determining the number of viable cells with a tetrazolium compound and by measuring lactate dehydrogenase (LDH) release, respectively. Virus titration and antiviral cytokines mRNA expression were evaluated by quantitative PCR. DON significantly affected the survival of noninfected cells in a dose dependent manner. However, DON concentrations between 140 and 280 significantly increased the survival of cells infected with PRRSV. These concentrations significantly decreased PRRSV replication by inducing a pro-inflammatory cytokines environment and an early activation of apoptosis, which in turn seem to interrupt viral replication. For the first time, this study showed that DON had significant effects on the survival of PRRSV infected cells and on virus replication, in a dose dependent manner.

Effect of dietary phytate and microbial phytase on mineral and trace element bioavailability- a literature review

Phytic acid (PA) is the main phosphorus storage compound in cereals, legumes and oil seeds. In human populations where phytate-rich cereals such as wheat, maize and rice are a staple food, phytate may lead to mineral and trace element deficiency. Zinc appears to be the trace element whose bioavailability is most influenced by PA. Furthermore, several studies in humans as well as in monogastric animals clearly indicate an inhibition of non-haem iron absorption at marginal iron supply due to phytic acid. In fact PA seems to be, at least partly, responsible for the low absorption efficiency and high incidence of iron deficiency anaemia evident in most developing countries, where largely vegetarian diets are consumed Microbial phytases have provided a realistic means of improving mineral availability from traditionally high-phytate diets. In fact it has been consistently shown that Aspergillus phytases significantly enhance the absorption of calcium, magnesium and zinc in pigs and rats. Furthermore there are a few studies in humans indicating an improvement of iron bioavailability due to microbial phytase.

Probiosis: concepts and prospects

The use of probiotics in animal and human feeding has been a subject of increasing interest both for the academia and the industry, mostly due to their potential positive effects on health and profitability. However, the knowledge on the composition of bacterial gastrointestinal communities in humans and animals, as well as its respective nutritional requirements, is far from being elucidated. Due to the ban of the use of antibiotic growth promoters in animal feeds, there has been an increasing interest on the utilization of probiotics to improve animal intestinal health under commercial settings. However, the possibility of horizontal transference of antibiotic-resistant genes between probiotic bacteria and pathogenic species has become a concern of poultry farmers and consumers around the world. Innovative ideas have emerged, such as the addition of essential oils, spices, and other plant extracts to feeds of monogastric animals to promote intestinal health. These natural compounds are considered ecologically adequate and safe for feeding purposes. This new reality will probably change the direction of research and of the use of additives in poultry production.

Avanços metodológicos na avaliação de alimentos e de exigências nutricionais para aves e suínos

A produção industrial de aves e suínos mostrou enormes avanços nos últimos anos, principalmente devido ao aumento dos conhecimentos na área de genética e de nutrição. Na área de avaliação dos alimentos e das exigências nutricionais de animais monogástricos, as melhoras se dão a passos largos, devido á seriedade com que os profissionais encaram a responsabilidade de fazer pesquisa de qualidade dentro e fora do país. Neste trabalho serão abordadas as metodologias que permitem melhorar a utilização dos alimentos de maneira mais eficiente e econômica. Serão citados alguns cuidados e procedimentos essenciais para executar adequadamente experimentos de desempenho com aves e suínos. Na atualidade, para a realização de experimentos com monogástricos, é necessário: definir claramente os objetivos, utilizar animais com peso inicial uniforme, usar número adequado de repetições e de animais por unidade experimental. Testes de médias devem ser usados para as variáveis qualitativas e quando a variável independente for quantitativa aplicar análise de regressão. O nível de significância utilizado (5, 7, 10%) pode variar conforme a importância econômica da característica estudada. A adequada condução dos ensaios, sejam de crescimento ou de digestibilidade, é fundamental para que as ferramentas apresentadas possam ter efetividade, resultando na melhora da produtividade e na redução dos custos e da excreção de nutrientes.

Determinação da temperatura para inativação da urease em amostras de casca e farelo de soja

s not degraded in the rumen but destroy ureasis. Soybean meal is one of the protein ingredients commonly used in formulations of animal feeds. In the diets of monogastric bran has high protein content provided by a greater separation of soybean hulls. For the ruminant protein value of the meal is lower with the inclusion of soybean hulls to reduce the level of protein. With the increased consumption of meal high protein, due to increases in production of pigs and poultry in recent years, increasing the availability of soybean hulls on the market, this is because soybean hulls is little understood in the feeding of monogastric . In this context, the main objective of this study was to determine the times and temperatures needed to ensure the effective inactivation of ureasis present in the bran and soybean hulls. According to the assessments, to make the determination of temperatures and times required for the inactivation of ureasis present in the bran and soybean hulls are 170 º C and 25 minutes for soybean meal and 140 ° C 10 minutes for soybean hulls, respectively

Pruning management of cassava for animal feeding: parameters of the root

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nutritional impact on health and performance in intensively reared rabbits

The present work summarizes research related to the definition of nutrient recommendations for feeds used in the intensive production of rabbit's meat. Fibre is the main chemical constituent of rabbit diets that typically contain 320 to 360 and 50 to 90 g/kg of insoluble and soluble fibre, respectively. Instead, the dietary contents of cereal grains (∼120 to 160 g/kg), fat (15 to 25 g/kg) and protein concentrates (150 to 180 g/kg) are usually low with respect to other intensively reared monogastric animals. Cell wall constituents are not well digested in rabbits, but this effect is compensated by its stimulus of gut motility, which leads to an increasing rate of passage of digesta, and allows achieving an elevated dry matter intake. A high feed consumption and an adequate balance in essential nutrients are required to sustain the elevated needs of high-productive rabbits measured either as reproductive yield, milk production or growth rate in the fattening period. Around weaning, pathologies occur in a context of incomplete development of the digestive physiology of young rabbits. The supply of balanced diets has also been related to the prevention of disorders by means of three mechanisms: (i) promoting a lower retention time of the digesta in the digestive tract through feeding fibre sources with optimal chemical and physical characteristics, (ii) restricting feed intake after weaning or (iii) causing a lower flow of easily available substrates into the fermentative area by modifying feed composition (e.g. by lowering protein and starch contents, increasing its digestibility or partially substituting insoluble with soluble fibre), or by delaying age at weaning. The alteration in the gut microbiota composition has been postulated as the possible primary cause of these pathologies.

Caracterización molecular del mutante riso 1508 de cebada : modificaciones transcripcionales y posttranscripcionales en el desarrollo de la semilla

La semilla es el principal órgano reproductivo de las plantas espermatofitas, permitiendo la dispersión de las poblaciones y asegurando su supervivencia gracias a su tolerancia a la desecación y a su capacidad para germinar bajo condiciones ambientales óptimas. El rendimiento y valor económico de los cereales, que constituyen la primera cosecha mundial, depende, en buena medida, de la eficacia con que se acumulan en la semilla sustancias de reserva: proteínas, carbohidratos y lípidos. El principal carbohidrato acumulado en la semilla de cebada es el almidón y la fracción mayoritaria de proteínas es la de las prolaminas (solubles en etanol al 70%); estas proteínas tienen muy bajo contenido en lisina, un aminoácido esencial en la dieta de animales monogástricos. Con el fin de mejorar el valor nutricional de la semilla de cebada, se han obtenido diferentes mutantes con un mayor contenido en este aminoácido. Riso 1508 es un mutante de cebada rico en lisina cuya mutación lys3a, de efectos pleiotrópicos, segrega como un único gen mendeliano. Entre otros, presenta una reducción drástica de la expresión de algunos genes que codifican proteínas de reserva de tipo prolamina, en concreto, presenta reducida la expresión de los genes que codifican B-, C- y ϒ-Hordeínas y del inhibidor de tripsina CMe, pero no tiene alterada la expresión del gen que codifica las D-Hordeínas. Este último gen carece en su promotor del motivo GLM (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), que es reconocido por factores transcripcionales bZIP. En este trabajo, el mutante de cebada Riso 1508 se ha utilizado como herramienta para profundizar en el conocimiento de la regulación génica en semillas durante las fases de la maduración y la germinación. Para ello, en una primera aproximación, se llevó a cabo un análisis transcriptómico comparando el genotipo mutante con el silvestre durante la maduración de la semilla. Además de confirmar variaciones en los genes que codifican proteínas de reserva, este análisis indicó que también estaban afectados los genes relacionados con metabolismo de carbohidratos. Por ello se decidió caracterizar la familia multigénica de sacarosas sintasa (SUSy) en cebada. Se anotaron dos nuevos genes, HvSs3 y HvSs4, cuya expresión se comparó con la de los genes HvSs1 y HvSs2, previamente descritos en el laboratorio. La expresión de los cuatro genes en tejidos diferentes y su respuesta a estreses abióticos se analizó mediante RT-qPCR. HvSs1 y HvSs2 se expresaron preferencialmente durante el desarrollo del endospermo, y HvSs1 también fue un tránscrito abundante durante la germinación. HvSs1 se indujo en hojas en condiciones de anoxia y HvSs3 por estrés hídrico, y ambos genes se indujeron por tratamientos de frío. La localización subcelular de las cuatro isoformas no fue sólo citoplásmica, sino que también se localizaron en zonas próximas a retículo endoplásmico y en la cara interna de la membrana plasmática; además, se observó una co-localización de HvSS1 con el marcador de mitocondrias. Estos datos sugieren un papel distinto aunque parcialmente solapante de las cuatro Sacarosa Sintasas de cebada, descritas hasta la fecha. Las cinéticas de expresión de los genes que codifican los TFs más importantes implicados en la regulación génica durante el desarrollo del endospermo de cebada, se analizaron por RT-qPCR en ambos genotipos, demostrando que los TFs de la clase DOF aparecieron desregulados durante todo el proceso en Riso 1508 comparado con el cv. Bomi, aunque también se observaron diferencias significativas en algunos de los que codifican bZIPs. Estudios previos indicaban que el ortólogo de BLZ2 en maíz, O2, se regula post-traduccionalmente mediante un mecanismo de fosforilación/defosforilación reversible, y que la forma defosforilada es la fisiológicamente activa. En este trabajo se demostró que BLZ2 está sujeto a este tipo de regulación y que la proteín-fosfatasa HvPP2C2 está implicada en el proceso. La interacción de HvPP2C2 y BLZ2 tiene lugar en el núcleo celular únicamente en presencia de 100 μM ABA. En el mutante Riso 1508, BLZ2 se encuentra en un estado hiperfosforilado tanto durante la maduración como durante la germinación de la semilla, lo que dificultaría la unión de BLZ2 a las secuencias GLM en los promotores de los genes que codifican B-, C-,y ϒ- Hordeínas y CMe. Summary The seed is the main reproductive organ of spermatophyte plants allowing the spread of populations and ensuring their survival through its desiccation tolerance and because of their ability to germinate under optimum environmental conditions. Yield and economic value of cereal crops, that constitute the first world crop, depend largely on the efficiency with which they accumulate in the seed reserve substances: proteins, carbohydrates and lipids. The main carbohydrate accumulated in the barley seed is starch and the major protein fraction is that of prolamins (soluble in 70% ethanol); these proteins have a very low lysine content, an essential amino-acid for the diet of monogastric animals. In order to improve the nutritional value of the barley seed, different mutants have been obtained with a higher content of this amino-acid. Riso 1508 is one lysine-rich mutant whose mutation (lys3a) segregates as a single Mendelian gene with pleiotropic effects, such as a drastic reduction of genes encoding the trypsin inhibitor CMe and the B-, C-and ϒ-hordeins, but has not altered the expression of the gene encoding the D-hordeins. This latter gene lacks in its promotor the GLM motif (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), that is recognised by bZIP transcription factors In this work we have used the barley mutant Riso 1508 as a tool for better understanding gene regulation in seeds during the maturation and germination phases. To this aim, a transcriptomic analysis was performed comparing wild and mutant genotypes during seed maturation. Besides confirming variations in the expression of genes encoding reserve proteins, this analysis indicated that some genes related with carbohydrate metabolism were also affected. It was therefore decided to characterize the multigene family of sucrose synthases (SUSy) in barley. Two new genes were annotated, HvSs3 and HvSs4, and its expression was compared with that of genes HvSs1 and HvSs2, previously described in our laboratory. The expression of the four genes in different tissues and in response to abiotic stresses was analyzed by RTqPCR. HvSs1 and HvSs2 were preferentially expressed during the development of the endosperm, and the HvSs1 transcript was also abundant upon germination. HvSs1 was induced in leaves by anoxic conditions, HvSs3 by water stress, and both genes were induced by cold treatments. The subcellular localization of all four isoforms was not only cytoplasmic, but they could be found along the endoplasmic reticulum and at the inner side of the cell membrane; HvSS1, was also associated with the mitochondrial marker. These data suggest a distinct but partially overlapping roles for the barley sucrose synthases, described so far. The expression kinetics of the genes encoding the most important TFs involved in gene regulation during barley endosperm development was analyzed by RT-qPCR in both genotypes. These data show that the genes encoding DOF TFs were mis-regulated throughout the process in Riso 1508, although significant differences were also found among some of those encoding bZIPs. Previous studies indicated that the BLZ2 orthologue in maize, O2, was post-translationally regulated by reversible phosphorylation/dephosphorylation and that the dephosphorylated protein is the physiologically active form. In this work we demostrate that BLZ2 is under a similar regulation and that the proteinphosphatase HvPP2C2 is implicated in the process. The interaction between HvPP2C2 and BLZ2 takes place in the cell nucleus only in the presence of 100 μM ABA. In the Riso 1508 mutant, BLZ2 is found in a hyperphosphorylated state in the maturation phase and upon seed germination; because of this, the BLZ2 binding to the GLM promoter sequences of genes encoding B-, C- y ϒ- Hordeins and CMe would be decreased in the mutant.

Biological response of broiler chickens fed peas (Pisum sativum L.) expressing the bean (Phaseolus vulgaris L.) alpha-amylase inhibitor transgene

The nutritive value of transgenic peas expressing an a-amylase inhibitor (alpha-Ail) was evaluated with broiler chickens. The effects of feeding transgenic peas on the development of visceral organs associated with digestion and nutrient absorption were also examined. The chemical composition of the conventional and the transgenic peas used in this study were similar. In the two feeding trials, that were conducted normal and transgenic peas were incorporated into a maize-soybean diet at concentrations up to 500 g kg(-1). The diets were balanced to contain similar levels of apparent metabolisable energy (AME) and amino acids. In the first trial, the birds were fed the diets from 3 to 17days post-hatching and with levels of transgenic peas at 250 g kg(-1) or greater there was a significant reduction in body weight but an increase in feed intake resulting in deceased feed conversion efficiency. In the second trial, in which the birds were fed diets containing 300 g kg(-1) transgenic peas until 40 days of age, growth performance was significantly reduced. It was also demonstrated that the ileal starch digestibility coefficient (0.80 vs 0.42) was significantly reduced in the birds fed transgenic peas. Determination of AME and ileal digestibility of amino acids in 5-week-old broilers demonstrated a significant reduction in AME (12.12 vs 5.08 MJ kg(-1) DM) in the birds fed the transgenic peas. The AME value recorded for transgenic peas reflected the lower starch digestibility of this line. Real digestion of protein and amino acids was unaffected by treatment. Expression of a-Ail in peas did not appear to affect bird health or the utilisation of dietary protein. However, the significant reduction in ileal digestion of starch in transgenic peas does reduce the utility of this feedstuff in monogastric diets where efficient energy utilisation is required. (c) 2006 Society of Chemical Industry.