Influencia de los métodos de conservación en las actividades enzimáticas de suelos forestales mediterráneos

Soil storage method may alter enzymatic activity being storage conditions of the soil samples prior to analysis decisive for the results. Studies made on freshly collected soils are generally preferred. However it is always not possible due to practical reasons since for example sampling is often restricted to short period of the year or because a great quantity of microbiological analyses must be made on time and by few people. On this context, soil storage methods are needed, being cold at 4°C the most widely used although sometimes alternative storage methods are also utilized. The aim of this study is to evaluate the effect of two alternative storage methods of soil samples (freezing at –20°C and air drying conservation methods) in comparison to cold at 4°C on the enzymatic activities (dehydrogenase, phosphatase, β-glucosidase and urease soil enzymes). Samples of two forest ecosystems (pine and holm oak forest stand) were taken in two different season of the year (winter and spring 2009). Results showed that enzymatic activities differed when freezing or air drying conservation methods were used in comparison with cold soil samples. Generally, alternative soil storage methods presented lower enzymatic activity than cold at 4°C. However, these changes depend on season and sampling location.El método de conservación del suelo utilizado puede alterar la actividad enzimática, siendo decisivas para los resultados, las condiciones de conservación previas a los análisis. Para los estudios es preferible realizar los análisis en muestras frescas y recién cogidas. Sin embargo, esto no es siempre posible por razones prácticas ya que las muestras se tienen que recoger en un corto periodo del año o porque los análisis los debe hacer poca gente y siempre en un determinado tiempo. En este contexto, se necesita un método de conservación, siendo el mantenimiento en el frigorífico a 4°C el método más usado, aunque existen otros métodos alternativos. El objetivo de este trabajo es el de evaluar el efecto de dos métodos alternativos de conservación (congelado a –20°C y secado a temperatura ambiente) sobre las actividades enzimáticas (deshidrogenasa, fosfatasa, β-glucosidasa y ureasa), en comparación con el mantenido en el frigorífico a 4°C. Las muestras se obtuvieron de dos zonas forestales diferentes (pinar y encinar) y en dos épocas distintas (primavera y verano de 2009). Los resultados mostraron diferencias en las actividades enzimáticas cuando se usó el congelado o secado al aire de las muestras de suelo en comparación con el mantenimiento en el frigorífico. De forma general, los métodos de conservación alternativos mostraron una menor actividad enzimática en las muestras de suelo analizadas. Sin embargo, estos cambios dependen de la época del año y la zona de muestreo

Food and nutrition security discursive frames in the context of the Spanish economic crisis

[EN] The recent economic crisis in Spain has provoked a number of food and nutrition security (FNS) impacts that have fuelled a fragmented landscape of food-related debates, although these have not taken the form of a coherent and allembracing public discussion on national food security. This paper is an inductive analysis of such debates, mostly based on media coverage, in an attempt to disentangle the different discourse frames on FNS in Spain, and the role played by the economic crisis in the shaping and underpinning of such frames. For this purpose a frame matrix was constructed, paying particular attention to the stakeholders governance claims in order to confront frame-specific problems. Results show how stakeholders have used elements of the crisis to underpin or reinterpret their core arguments. In addition, governance claims have concentrated on the malfunctioning of the food chain, mostly due to unequal bargaining power, and the way this could be putting at risk crucial components of the food system and future FNS. Also, the short-termism of most stakeholders approaches obscures the long run threats to the Spanish food system.This research is part of the project "Assessment of the impact of global drivers of change on Europe's food security" (TRANSMANGO), granted by the EU under 7th Framework Programme, theme KBBE.2013.2.5-01, Grant agreement no: 613532. Authors gratefully appreciate the comments and suggestions made by the three anonymous reviewers and the editors of the Journal.OOrtiz Miranda, D.; Moreno Pérez, OM.; Arnalte Alegre, EV. (2016). Food and nutrition security discursive frames in the context of the Spanish economic crisis. Food Security. 8(3):665-677. https://doi.org/10.1007/s12571-016-0574-xS66567783Antentas, J. M., & Vivas, E. (2014). 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National Report. Italy. WP2 report Transmango EU-FP7.GREENPEACE (2014). Alimentos bajo amenaza. Valor económico de la polinización y vulnerabilidad de la agricultura española ante el declive de las abejas y otros polinizadores. Madrid: Greenpeace.Hertel, T. W. (2015). The challenges of sustainably feeding a growing planet. Food Security, 7, 185–198.Hispacoop (2012). Estudio sobre desperdicio de alimentos en los hogares. Madrid: Confederación Española de Cooperativas de Consumidores y Usuarios.Lang, T., & Heasman, M. (2004). Food wars. The global battle for mouths, Minds and Markets. London: Earthscan.Lawrence, G., & McMichael, P. (2012). The question of food security. International Journal of Sociology of Agriculture & Food, 19(2), 135–142.MAGRAMA. (2014). La balanza commercial agroalimentaria en 2013. Madrid.Maye, D., & Kirwan, J. (2013). Food security: a fractured consensus. Journal of Rural Studies, 29, 1–6.McKeon, N. (2015). Food security governance: empowering communities, regulating corporations. New York: Routledge.Mooney, P. H., & Hunt, S. A. (2009). Food security: the elaboration of contested claims to a consensus frame. Rural Sociology, 74(4), 469–497.Moreno, O. (2013). Reproducing productivism in Spanish agricultural systems. In D. Ortiz, A. Moragues, & E. Arnalte (Eds.), Agriculture in Mediterranean Europe: between old and new paradigms. Research in rural sociology and development (pp. 121–147). Bingley: Emerald Group Publishing Limited.Muñoz, C., & Sosvilla, S. (2014). Alimentamos el futuro. Informe económico 2013. Madrid: FIAB.OECC - Oficina Española de Cambio Climático (2006). Plan Nacional de Adaptación al Cambio Climático. Madrid.OECC-UCLM (2005). Evaluación Preliminar de los Impactos en España por Efecto del Cambio Climático. Madrid: MAGRAMA.Pangaribowo, E. H., Gerber, N., & Torero, M. (2013). Food and Nutrition Security Indicators: A Review. ZEF working paper no 108. Bonn: ZEF.Sáez-Almendros, S., Obrador, B., Bach-Faig, A., & Serra-Majem, L. (2013). Environmental footprints of Mediterranean versus Western dietary patterns: beyond the health benefits of the Mediterranean diet. Environmental Health, 12, 118.Shepherd, B. (2012). Thinking critically about food security. Security Dialogue, 43(3), 195–212.Sineiro, F. (2012). Situación actual, retos y perspectivas del vacuno de leche. Available at http://www.eumedia.es/portales/files/documentos/2-FSineiro.pdf . Accessed 19 Nov 2015.Van Gorp, B. (2007). The constructionist approach to framing: bringing culture back. Journal of Communication, 57, 60–78.Van Gorp, B., & van der Goot, M. J. (2012). Sustainable food and agriculture: Stakeholder’s frames. Communication, Culture & Critique, 5, 127–148.Vidal, R., Moliner, E., Pikula, A., Mena-Nieto, A., & Ortega, A. (2015). Comparison of the carbon footprint of different patient diets in a Spanish hospital. Journal of Health Services Research & Policy, 20(1), 39–44

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7473G>A (p.=) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of 3 mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that 4 of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease.info:eu-repo/semantics/publishedVersio

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to the identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7437G>A) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of three mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that four of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease. clinicaltrials.gov identifier:02869074

Evaluation of the antimicrobial effect of a probiotic mixture against Cronobactersakazakii and Salmonellaenterica in infant formulas

Cronobacter sakazakii y Salmonella enterica son considerados patógenos capaces de causar sepsis y menin-gitis en lactantes. Se han descrito casos de infección por estos patógenos asociados al consumo de preparados para lactantes contaminados.La inclusión de probióticos en los preparados, además de su papel en la modulación de la microbiota intestinal, podría ser útil para contrarrestar la contaminación de microorganismos patógenos de estos productos una vez reconstituidos y, por lo tanto, reducir el riesgo de infección. El objetivo de esta investigación fue evaluar el efecto antimicrobiano de una mezcla de probióticos (Lactobacillus rhamnosus y Bifidobacterium infantis) frente a Cronobacter sakazakii y Salmonella enterica en preparados reconstituidos para lactantes. Las muestras (con o sin probióticos) se inocularon con C. sakazakii o S. enterica y se incubaron a 30 o 40ºC durante 72 horas. Los resultados muestran que la presencia de probióticos en los preparados lácteos no tiene un marcado efecto en el crecimiento de los patógenos. Sin embargo sí se ve potenciado su efecto inhibitorio frente a C. saka-zakii por la interacción entre la temperatura de incubación y los probióticos. Los probióticos empleados fueron viables durante todo el período de incubación, por lo que el agotamiento de los nutrientes se descartó como posi-ble mecanismo de inhibición. La disminución del pH en los preparados lácteos inoculados no explica de manera satisfactoria un comportamiento tan diferente entre ambos patógenos. Por lo tanto, se sugiere como posible me-canismo inhibidor la liberación de bacteriocinas específicas con acción contra C. sakazakii pero sin efecto contra Salmonella. Se deben realizar estudios adicionales para probar esta hipótesis.Cronobacter sakazakii and Salmonella enterica have been implicated in outbreaks causing sepsis and me-ningitis in infants. Several infection cases have been associated with the consumption of contaminated powde-red infant formulae. Besides its role in modulating gut microbiota, the inclusion of probiotics in infant formulae may be useful to counteract pathogen contamination in these products and thus to reduce the risk of infection. The aim of this research was to evaluate the antimicrobial effect of a mixture of the probiotics Lactobacillus rhamnosus and Bifidobacterium infantis against Cronobacter sakazakii and Salmonella enterica in reconstitu-ted follow-on formulae. For that, infant formulae (with or without probiotics) were inoculated with either C. sakazakii or S. enterica and incubated at 30 or 40ºC for 72 hours. Overall, the results showed the presence of probiotics inhibiting the growth of C. sakazakii together with incubation temperature (interaction between both factors). Probiotics were viable throughout the incubation period so that nutrient exhaustion could be ruled out as the mechanism of C. sakazakii inhibition. Thus, the acidification of media observed during the study was hypothesized to be responsible for pathogen growth inhibition. However, since in the case of formulae inoculated with Salmonella the behavior of probiotics and pH values was similar to C. sakazakii, the decrease of pH does not satisfactorily explain such a different behavior. So, it is suggested that the production of specific bacteriocins by probiotics against C. sakazakii but not against Salmonellla could be the underlying inhibitory mechanism. Further studies should be carried out to test this hypothesis

Workflow for microbiological diagnosis of bacterial gastroenteritis combining a molecular assay as first-line with reflective stool culture

<p>Conventional microbiological methods for bacterial enteric disease diagnosis are time-consuming, labour intensive and provide low sensitivity. The aim of this study was to evaluate the results of a new diagnosis strategy which replaces traditional stool culture with a molecular detection using the BD MAX™ System (BD Life Sciences, Sparks, Maryland, United States) as first-line assay together with reflective culture. A total of 1.590 specimens were prospectively requested for stool culture. The molecular detection included the BD MAX enteric bacterial panel together with the BD MAX extended enteric bacterial panel (BDM GIP) performed simultaneously on the same stool specimen. In 18.8% of specimens (176 of the 936 valid samples) there was one or more than one target positive with the following percent positivity: 9.7% Campylobacter spp., 5.7% Salmonella spp., 1.3% Shiga toxin genes (stx1/stx2), 1.2% Shigella spp./enteroinvasive Escherichia coli (EIEC), 1% Yersinia enterocolitica, 1% Vibrio spp. (V. vulnificus/V. parahaemolyticus/V. cholerae), 0.3% Plesiomonas shigelloides, and 0.2% Enterotoxigenic E. coli (ETEC) enterotoxin LT/ST genes. Positive reflective stool culture noted a correlation of 69.5% with the molecular test, missing 23.9% and 15.4% in the cases of Campylobacter spp., and Salmonella spp., respectively. In conclusion, this clinical study demonstrated very good performance of the BDM GIP. The performance and ease of use may provide advantages to many laboratories, improving the detection of bacterial stool pathogens and time to reporting results.</p&gt

Evaluación del efecto antimicrobiano de una mezcla de probióticos frente a Cronobacter sakazakii y Salmonella enterica en preparados para lactantes reconstituidos para lactantes

Cronobacter sakazakii and Salmonella enterica have been implicated in outbreaks causing sepsis and meningitis in infants. Several infection cases have been associated with the consumption of contaminated powdered infant formulae. Besides its role in modulating gut microbiota, the inclusion of probiotics in infant formulae may be useful to counteract pathogen contamination in these products and thus to reduce the risk of infection. The aim of this research was to evaluate the antimicrobial effect of a mixture of the probiotics Lactobacillus rhamnosus and Bifidobacterium infantis against Cronobacter sakazakii and Salmonella enterica in reconstituted follow-on formulae. For that, infant formulae (with or without probiotics) were inoculated with either C. sakazakii or S. enterica and incubated at 30 or 40ºC for 72 hours. Overall, the results showed the presence of probiotics inhibiting the growth of C. sakazakii together with incubation temperature (interaction between both factors). Probiotics were viable throughout the incubation period so that nutrient exhaustion could be ruled out as the mechanism of C. sakazakii inhibition. Thus, the acidification of media observed during the study was hypothesized to be responsible for pathogen growth inhibition. However, since in the case of formulae inoculated with Salmonella the behavior of probiotics and pH values was similar to C. sakazakii, the decrease of pH does not satisfactorily explain such a different behavior. So, it is suggested that the production of specific bacteriocins by probiotics against C. sakazakii but not against Salmonellla could be the underlying inhibitory mechanism. Further studies should be carried out to test this hypothesis.Cronobacter sakazakii y Salmonella enterica son considerados patógenos capaces de causar sepsis y meningitis en lactantes. Se han descrito casos de infección por estos patógenos asociados al consumo de preparados para lactantes contaminados. La inclusión de probióticos en los preparados, además de su papel en la modulación de la microbiota intestinal, podría ser útil para contrarrestar la contaminación de microorganismos patógenos de estos productos una vez reconstituidos y, por lo tanto, reducir el riesgo de infección. El objetivo de esta investigación fue evaluar el efecto antimicrobiano de una mezcla de probióticos (Lactobacillus rhamnosus y Bifidobacterium infantis) frente a Cronobacter sakazakii y Salmonella enterica en preparados reconstituidos para lactantes. Las muestras (con o sin probióticos) se inocularon con C. sakazakii o S. enterica y se incubaron a 30 o 40ºC durante 72 horas. Los resultados muestran que la presencia de probióticos en los preparados lácteos no tiene un marcado efecto en el crecimiento de los patógenos. Sin embargo sí se ve potenciado su efecto inhibitorio frente a C. sakazakii por la interacción entre la temperatura de incubación y los probióticos. Los probióticos empleados fueron viables durante todo el período de incubación, por lo que el agotamiento de los nutrientes se descartó como posible mecanismo de inhibición. La disminución del pH en los preparados lácteos inoculados no explica de manera satisfactoria un comportamiento tan diferente entre ambos patógenos. Por lo tanto, se sugiere como posible mecanismo inhibidor la liberación de bacteriocinas específicas con acción contra C. sakazakii pero sin efecto contra Salmonella. Se deben realizar estudios adicionales para probar esta hipótesis.

Long term forest management drives drought resilience in Mediterranean black pine forest

Spanish black pine showed greater resilience and resistance, but generally lower recovery to drought events in managed than in unmanaged forest stands under Mediterranean humid climate. Abstract Drought negative efects on forest ecosystems are projected to increase under global warming all over the world. In this context, forest management can be an efective option for reducing drought impacts and increasing tree growth stability to extreme drought events. Here, we aim to evaluate black pine (Pinus nigra subsp. salzmannii) growth response to climatic variability and drought events in managed and unmanaged stands under similar Mediterranean climatic conditions. Drought events were identifed using long-term climatic data, and basal area increments were calculated for 100–120-year old trees cored in managed and unmanaged plots. Results showed that tree size, temperature, and the interaction between management treatment and water availability signifcantly infuenced tree growth. Basal area increment was reduced in response to the 1983, 1991, 1994‒1995, 1999–2000 and 2005 drought events. Trees in managed plots showed lower growth reductions in response to drought than those located in unmanaged plots, probably experiencing higher competition for soil water, whereas the reverse happened under wet climate conditions. Black pines showed greater resilience and resistance, but generally lower recovery to drought events in managed than in unmanaged stands. Our results suggest that forest management enhances drought tolerance in black pine stands, which may help to ameliorate the negative impacts of global warming across Mediterranean forest ecosystems