Medidas de manejo del síndrome reproductor y respiratorio porcino (PRRS) basadas en su diagnostico molecular

El síndrome reproductor y respiratorio porcino (PRRS) es una de las enfermedades infecciosas más importantes por el impacto económico que supone para la industria porcina nacional e internacional. Está producida por un virus ARN cuyas principales características son la variabilidad genética y antigénica, sus propiedades inmunomoduladoras y su capacidad para inducir infecciones persistentes. Han sido descritos dos genotipos principales del virus de PRRS, el europeo (EU) y el americano (NA) y últimamente, un tercer genotipo presente solo en Asia La comparación de sus secuencias ha mostrado diferencias genéticas significativas entre ellos, lo que contribuye a que la vacunación sea poco efectiva. Uno de los métodos más empleados para el diagnostico de PRRS es la RT PCR que, acompañada de secuenciación, nos permite determinar el genotipo viral. En los últimos años se han empezado a desarrollar nuevas técnicas de RT-PCR en tiempo real que suponen una mejora sobre las convencionales debido a que presentan ventajas como su mayor sensibilidad, la posibilidad de cuantificar la carga viral o el ahorro de tiempo y material, mejorando el control de la enfermedad. En este trabajo queremos hacer patentes las ventajas y aplicaciones de la RT-PCR en tiempo real para el diagnóstico y el control del virus del Síndrome Respiratorio y Reproductor Porcino.The porcine reproductive and respiratory syndrome (PRRS) is one of the most important infectious diseases in swine national and international industry because of its economic impact. It is caused by a single stranded RNA virus which its main characteristics are: genetic and antigenic variability, immune modulation properties and its capacity to produce persistent infections. There have been described two main genotypes, the North American (NA) and the European (EU) and lately, a third one still limited to Asia. The comparison of its sequences has shown important genetic differences between them, what contributes to the failure of the existing vaccines against it. One of the most used diagnostic methods is the RT-PCR which complemented with sequencing, leads to determinate the viral genotype. In the last years, new RT-PCRs in real time have been developed because they present advantages as higher sensitivity, quantification of viral load or saving on material and time, improving the control of the disease. In this job, we would like to show the advantages and applications of the real time RTPCR for the diagnostic and control of the PRRS virus

Pancreatic Duct Cells in Human Islet Cell Preparations Are a Source of Angiogenic Cytokines Interleukin-8 and Vascular Endothelial Growth Factor

OBJECTIVE—Engraftment and function of human islet cell implants is considered to be dependent on their rapid and adequate revascularization. Studies with rodent islet grafts have shown that vascular endothelial growth factor (VEGF) expression by β-cells can promote this process. The present work examines whether human islet preparations produce VEGF as well as interleukin (IL)-8, another angiogenic protein, and assesses the role of contaminating duct cells in VEGF and IL-8–mediated angiogenesis

Chromatographic Examinations of Tea's Protection Against Lipid Oxidative Modifications

Ethanol metabolism is accompanied by generation of free radicals that damage cell components, especially lipids. The present study was designed to investigate the efficacy of the preventive effect of black tea on the lipid oxidative modifications in different tissues (plasma, liver, brain, kidney, stomach, lung, intestine, and spleen) of 12-month-old rats chronically intoxicated with ethanol. Ethanol intoxication caused changes in the level/activity of antioxidants that led to the significant increase in the level of lipid oxidative modification products. Oxidative modifications were estimated by measuring lipid hydroperoxides, malondialdehyde, and 4-hydroxynonenal by high-performance liquid chromatography (HPLC) and by spectrophotometric determination of conjugated dienes. These lipid-modification marker levels were increased in almost all examined tissues (3%–71%) after ethanol intoxication. Described changes were in accordance with the liver level of the most often used marker of arachidonic acid oxidation, isoprostane (8-isoPGF2α), determined by the LC/MS system. Administration of black tea to ethanol-intoxicated rats remarkably prevents the significant increase (by about 15%–42%) in concentrations of all measured parameters regarding all examined tissues, but especially the plasma, liver, brain, stomach, and spleen. The preventive effect of black tea in the other organs (kidney, lung, intestine) caused a decrease in examined markers in a smaller degree (by about 7%–28%). To determine in the liver the major constituents of black tea mainly responsible for antioxidative action such as catechins and theaflavins, which were absorbed in organism, the present study indicates their protective effect against ethanol-induced oxidative modifications of lipids

Age Related Changes in NAD+ Metabolism Oxidative Stress and Sirt1 Activity in Wistar Rats

The cofactor nicotinamide adenine dinucleotide (NAD+) has emerged as a key regulator of metabolism, stress resistance and longevity. Apart from its role as an important redox carrier, NAD+ also serves as the sole substrate for NAD-dependent enzymes, including poly(ADP-ribose) polymerase (PARP), an important DNA nick sensor, and NAD-dependent histone deacetylases, Sirtuins which play an important role in a wide variety of processes, including senescence, apoptosis, differentiation, and aging. We examined the effect of aging on intracellular NAD+ metabolism in the whole heart, lung, liver and kidney of female wistar rats. Our results are the first to show a significant decline in intracellular NAD+ levels and NAD∶NADH ratio in all organs by middle age (i.e.12 months) compared to young (i.e. 3 month old) rats. These changes in [NAD(H)] occurred in parallel with an increase in lipid peroxidation and protein carbonyls (o- and m- tyrosine) formation and decline in total antioxidant capacity in these organs. An age dependent increase in DNA damage (phosphorylated H2AX) was also observed in these same organs. Decreased Sirt1 activity and increased acetylated p53 were observed in organ tissues in parallel with the drop in NAD+ and moderate over-expression of Sirt1 protein. Reduced mitochondrial activity of complex I–IV was also observed in aging animals, impacting both redox status and ATP production. The strong positive correlation observed between DNA damage associated NAD+ depletion and Sirt1 activity suggests that adequate NAD+ concentrations may be an important longevity assurance factor

Comunicación corta. Primera detección del virus de la parálisis aguda israelita (IAPV) en España

Spanish bee samples were analyzed for the presence of Israel Acute Paralysis Virus (IAPV). Some of these samples were collected from colonies presenting compatible symptoms with the colony collapse disorder (CCD, 240 out of 484) and the rest were asymptomatic. Only one of these samples was diagnosed as positive to IAPV by employing a one step RT-PCR that targets the ORF 2 of the IAPV genome. Specificity of the RT-PCR assay was evaluated by sequence analysis of size specific amplification products. IAPV nucleotide sequences already published in GeneBank were used to construct a phylogenetic tree that included the new Spanish IAPV sequence (FJ821506). They segregated in two main lineages and the Spanish isolate was mainly related with the American ones. As IAPV was detected in Spain in a very low frequency, no causal relation between IAPV and CCD in Spain was found.Se analizaron muestras de abejas españolas para detectar el virus de la parálisis aguda israelita (IAPV). Parte de las muestras (240 de 484) procedían de colmenas que mostraban síntomas compatibles con el síndrome de despoblamiento (SDC), el resto de las muestras eran asintomáticas. Solo una de las muestras fue diagnosticada como positiva frente a IAPV utilizando la técnica RT-PCR, que amplifica una parte del ORF 2 del genoma viral. La especificidad de la prueba fue confirmada por secuenciación del producto amplificado que presentaba el tamaño específico. Se emplearon secuencias de IAPV ya publicadas en GeneBank para construir un árbol filogenético que incluye la secuencia de IAPV detectada en muestras españolas (FJ821506). Se observaron dos grupos principales y la muestra española está agrupada con varias americanas. Debido a que en España IAPV fue detectado en muy baja frecuencia, no se encuentra relación causal entre el SDC y la presencia de IAPV

Metagenomic Detection of Viral Pathogens in Spanish Honeybees: Co- Infection by Aphid Lethal Paralysis, Israel Acute Paralysis and Lake Sinai Viruses

The situation in Europe concerning honeybees has in recent years become increasingly aggravated with steady decline in populations and/or catastrophic winter losses. This has largely been attributed to the occurrence of a variety of known and "unknown", emerging novel diseases. Previous studies have demonstrated that colonies often can harbour more than one pathogen, making identification of etiological agents with classical methods difficult. By employing an unbiased metagenomic approach, which allows the detection of both unexpected and previously unknown infectious agents, the detection of three viruses, Aphid Lethal Paralysis Virus (ALPV), Israel Acute Paralysis Virus (IAPV), and Lake Sinai Virus (LSV), in honeybees from Spain is reported in this article. The existence of a subgroup of ALPV with the ability to infect bees was only recently reported and this is the first identification of such a strain in Europe. Similarly, LSV appear to be a still unclassified group of viruses with unclear impact on colony health and these viruses have not previously been identified outside of the United States. Furthermore, our study also reveals that these bees carried a plant virus, Turnip Ringspot Virus (TuRSV), potentially serving as important vector organisms. Taken together, these results demonstrate the new possibilities opened up by high-throughput sequencing and metagenomic analysis to study emerging new diseases in domestic and wild animal populations, including honeybees

Presencia, primera cuantificación y filogenia del virus de la parálisis aguda israelita (IAPV) de las abejas en Andalucía (España)

This study aimed to assess the possible relationship between the presence of Israeli acute paralysis virus (IAPV) of honeybees and disease symptoms development at the colony level, to describe the IAPV load in field colonies and to illustrate phylogenetic relationships between IAPV isolates in Andalusia (Spain). Presence and load of IAPV was studied in 96 colonies from all provinces in Andalusia. Epidemiological surveys were performed in all the colonies to assess their sanitary status. IAPV was found in 13.5% of the sampled colonies, and no association was observed between the presence of IAPV and disease symptoms at the colony level. An average IAPV load was established in 4.9•105 genome equivalent copies per bee. Phylogenetic analysis revealed that Andalusian isolates belong to a different lineage to a previously described isolate found in Valencia (2010). The results of this study will help us understand the epidemiology and effect of IAPV on Spanish colonies.Los objetivos de este trabajo son estudiar la posible relación entre la presencia del virus de la parálisis aguda israelita (IAPV) de las abejas y la aparición de síntomas a nivel de la colonia, así como describir la carga viral presente en colmenas comerciales e ilustrar las relaciones filogenéticas entre los diferentes aislados de IAPV en la región de Andalucía (España). Andalucía fue elegida para realizar un amplio muestreo de colonias de abejas ya que se trata de la región española con mayor censo de colmenas y la segunda en producción de miel. La presencia y carga de IAPV se estudió en 96 colmenas de todas las provincias de Andalucía. Se realizaron encuestas epidemiológicas en cada colonia para estudiar su estado sanitario. IAPV fue encontrado en el 13.5% de las colmenas muestreadas y no se estableció asociación entre la presencia de IAPV y la presencia de síntomas de enfermedad a nivel de la colonia. La carga viral media de IAPV se estableció en 4.9x105 copias equivalentes de genoma por abeja. El análisis filogenético reveló que los aislados de Andalucía pertenecen a un linaje distinto al del aislado previamente descrito en Valencia en 2010. Los resultados de este estudio ayudarán a entender la epidemiología y efecto de IAPV en las colmenas españolas