Identification and characterization of fungi associated with esca in vineyards of the Comunidad Valenciana (Spain)

Abstract Grapevines sampled in the Comunidad Valenciana in Spain were examined for esca symptoms, and the different vine parts (trunk, cane and young vine) were surveyed for the presence of esca-related fungi. The fungal species most frequently identified were the mitosporic fungi Diplodia mutila (Dmu), Phaeoacremonium aleophilum (Pal), Phaeomoniella chlamydospora (Pch) and Phomopsis viticola (Pvi) and the basidiomycetes Fomitiporia mediterranea (Fm) and Stereum hirsutum (Shi). PCR and sequencing using the universal pair primers ITS1-ITS4 confirmed the identification, these techniques proved very useful, particularly in cases where fruiting was not evident and/or pure cultures did not display enough diagnostic traits. Dmu and both basidiomycetes were the most frequent fungi associated with esca in this region. Dmu was the only fungal species found in all vine parts studied, being significantly more present in canes (82%) than in trunks (58%) or shoots (28%). Moreover, both basidiomycetes (50% Fm and 56% Shi) and Pch (32%) were most frequently found in trunks. Additional key words: Basidiomycetes, grapevine, ITS, mitosporic fungi, molecular characterization, morphological characteristics, rDNA, Vitis vinifera. Resumen Identificación y caracterización de hongos asociados a la yesca en viñedos de la Comunidad Valenciana (España) Se muestrearon viñas en la Comunidad Valenciana y se examinaron para detectar síntomas de yesca, y analizar la presencia de hongos relacionados con dicha enfermedad en las diferentes partes de la planta (tronco, caña y sarmiento). Las especies fúngicas identificadas de forma más frecuente fueron los hongos mitospóricos Diplodia mutila (Dmu), Phaeoacremonium aleophilum (Pal), Phaeomoniella chlamydospora (Pch) y Phomopsis viticola (Pvi) y los basidiomicetes Fomitiporia mediterranea (Fm) y Stereum hirsutum (Shi). Dicha identificación fue confirmada mediante PCR y secuenciación con la pareja de primers universal ITS1-ITS4, siendo especialmente útil en aquellos casos donde la fructificación no fue evidente y/o en los cultivos puros que no mostraron suficientes caracteres diagnósticos. Dmu, junto con ambos basidiomicetes, fueron los hongos más encontrados asociados a la yesca en esta región. Dmu fue la única especie fúngica presente en todas las partes de la viña estudiadas, con una incidencia significativa en cañas (82%) comparada con los troncos (58%) y los sarmientos (28%). Además, se encontraron de forma mayoritaria en el tronco los basidiomicetes (50% Fm y 56% Shi), así como Pch (32%)

Comparison of two techniques for the morphometry study on gilthead seabream (Sparus aurata) spermatozoa and evaluation of changes induced by cryopreservation

[EN] The development of powerful software has made possible spermatozoa morphology studies. However, some problems have emerged in relation to protocol standardization to compare results from different laboratories. This study was carried out to compare two techniques commonly used (staining vs phase contrast technique) for the morphometry study of gilthead sea bream spermatozoa using an integrated sperm analysis system (ISAS). Spermatozoa morphometry values were significantly affected by the technique used, and phase contrast technique was found to be the more accurate method, showing lower coefficients of variation on spermatozoa morphometry parameters measurements. Moreover, it has been shown that cryopreservation process produces damage in gilthead sea bream spermatozoa, causing negative effects in sperm parameters as spermatozoa morphometry (a decrease in cell volume), motility (from 95 to 68% motile cells) and viability (from 95 to 87% of live cells), being the addition of freezing medium containing cryoprotectant (DMSO) an important factor that caused the morphometry changes. (C) 2012 Elsevier Inc. All rights reserved.This work was financed by the Spanish Ministry of Science and Innovation (MICINN; AGL2007-64040-C03-00, Project SELECTBREAM). V. Gallego and I. Mazzeo were supported by predoctoral scholarships financed by the Spanish MICINN and Generalitat Valenciana, respectively. D.S. Penaranda had a postdoctoral grant from UPV.Gallego Albiach, V.; Peñaranda, D.; Marco Jiménez, F.; Mazzeo, I.; Pérez Igualada, LM.; Asturiano Nemesio, JF. (2012). Comparison of two techniques for the morphometry study on gilthead seabream (Sparus aurata) spermatozoa and evaluation of changes induced by cryopreservation. Theriogenology. 77(6):1078-1087. https://doi.org/10.1016/j.theriogenology.2011.10.010S1078108777

Nutritional innovations in superior European sea bass (Dicentrarchus labrax) genotypes: implications on fish performance and feed utilization.

The sustainable development of the aquaculture industry relies on the use of alternative conventional and emergent raw materials that contributes to a circular economy and to reduce the dependency on fish meals and fish oils coming from oceanic fish populations. Additionally, the genetic selection of farmed fish that can display higher growth and feed utilization when fed alternative feeds, is pointed out to be a complementary valuable tool to facilitate the implementation of circular economy approaches. The main purpose of the present study was to determine the effectiveness of genetic selection for growth in European sea bass, in response to a challenge with an alternative diet that aimed to partially replaced fishmeal (FM) by poultry meal (PM) and totally replace fish oil (FO) by a blend of poultry oil (PO) with a novel microalgae oil. The two families of fish juveniles were obtained by in vitro fertilization of selected for a multi-trait including high growth (genetically selected, GS) or nonselected (wild type, WT) broodstocks and then were nutritionally challenged with a control diet that mirrored a standard commercial diet with fishmeal (20%) and fish oil (7%), or a Future diet that partially replaced the FM by PM and totally replaced the FO by a blend of rapeseed oil, PO, and a novel DHA rich-algal oil. From the second month of feeding until the end of the trial, European sea bass that was selected since for 7 generations performed better in terms of growth than the wild-type genotype, possibly related with an apparent favored feed and nutrient utilization. Furthermore, selection decreased the perivisceral fat and increased the nutritional value of flesh by increasing DHA (in g/ 100 g flesh) and ARA contents. In contrast, the dietary treatment showed little effect on fish growth performance, denoting the successful partial replacement of FM by PM and the total replacement of FO by a blend of poultry oil and an emergent microalgal oil. However, Future diet tended to reduce the ADCs of some amino acids, as well as showed an additive effect to genotype in increasing the n-3 PUFA of flesh. Altogether, our data demonstrate that multi-trait genetic selection of European sea bass improve fish plasticity to cope with the variations of ingredients in alternative feeds with low FM/FO

Impact of non-adherence to radiotherapy on 1-year survival in cancer patients in Catalonia, Spain

This study aims to assess the effects of non-adherence to external beam radiation therapy in cancer patients receiving treatment with a curative. This retrospective cohort study collected health records data for all cancer patients treated with external beam radiotherapy with curative intent in 2016 in Catalonia, Spain. Adherence was defined as having received at least 90% of the total dose prescribed. A logistic regression model was used to assess factors related to non-adherence, and its association with one-year survival was evaluated using Cox regression. The final sample included 8721 patients (mean age 63.6 years): breast cancer was the most common tumour site (38.1%), followed by prostate and colon/rectum. Treatment interruptions prolonged the total duration of therapy in 70.7% of the patients, and 1.0% were non-adherent. Non-adherence was associated with advanced age, female gender, and some localization of primary tumour (head and neck, urinary bladder, and haematological cancers). The risk of death in non-adherent patients was higher than in adherent patients (hazard ratio [HR] 1.63, 95% confidence interval 0.97-2.74), after adjusting for the potential confounding effect of age, gender, tumour site and comorbidity. Non-adherence to radiotherapy, as measured by the received dose, is very low in our setting, and it may have an impact on one-year survival

El abastecimiento con agua subterránea a la Colonia Clunia Sulpicia (Hispania Citerior Tarraconensis).

Hace 20 siglos, una autoridad romana fundó una ciudad en lo alto de un cerro testigo donde el hallazgo, por habilidad o casualidad, de un pequeño acuífero permitió el abastecimiento de agua. En época romana se perforaron pozos, de los que diecinueve llegaron a una cavidad natural. También se abrió una galería horizontal, para captar agua, y un drenaje en el teatro para eliminar aguas que molestaban. En algún momento, por razones naturales o artificiales, el nivel freático bajó y varios pozos se secaron, obligando al descenso de trabajadores y otras personas. Además de hacer obras complementarias dejaron abundantes grafitis y varias figuritas en barro en la cueva. La ciudad no sobrevive a la Edad Media, los pozos son cegados y las ruinas sufren un proceso de expolio por las localidades vecinas. Recuperada su memoria en el siglo XVIII, en 1908 se redescubre cueva Román, una antigua galería de captación que da acceso a la cueva natural. Esta es explorada y topografiada por el grupo Espeleológico Ribereño desde 1980. A partir de 2013, el Grupo de Tecnologías en Entornos hostiles, de la Universidad de Zaragoza, realiza radiolocalizaciones sistemáticas para la apertura de un acceso directo a la cavidad. Además, se han hecho reconocimientos de inscripciones con escaneos de la cavidad y detalles específicos así como la instalación de dos estaciones de monitorización, exterior e interior, para analizar el funcionamiento hídrico del sistema y la evolución interior del contenido en CO2 del aire de la cavidad. La evolución del nivel piezométrico presenta oscilaciones relativamente importantes y se detecta un retraso de dos meses entre los momentos de fuertes lluvias y el ascenso. El contenido en CO2 presenta una doble periodicidad, anual y diaria. Twenty centuries ago, a Roman authority founded a city on top of an inselberg where the discovery, by skill or chance, of a small aquifer which allowed access to a water supply for the city. In Roman times wells were drilled, of which nineteen reached a natural cavity. A horizontal gallery for water supply was also opened up, and a drainage conduct in the theatre was made to eliminate storm water. At some point, for natural or artificial reasons, the water table dropped and several wells dried up, forcing the descent of workers and other people to do additional work and to carve abundant graffiti and several clay figurines. The city did not survive the Middle Ages, the wells became cesspits and the ruins were plundered by neighbouring villages. It recovered its memory in the eighteenth century, in 1908 a Roman cave was rediscovered, the old water supply gallery, which gives access to the natural cave. This has been explored and surveyed by the Grupo Espeleológico Ribereño since 1980. From 2013, the Grupo de Technologías in Entornos Hostiles (University of Zaragoza) has carried out systematic radiolocation for the opening up of a direct access to the cavity. In addition, recognition of inscriptions with cavity scans and specific details have been made as well as the installation of two monitoring stations, both inside and outside the cavity to analyze the water performance of the system and the internal evolution of the CO2 content of the air in the cavity. The evolution of the piezometric level presents relatively important oscillations and a delay of two months is detected between the moments of heavy rains and the increase of water levels in the cavity. The CO2 content has a double periodicity, annual and daily

Sperm motility parameters and spermatozoa morphometric characterization in marine species: a study of swimmer and sessile species

[EN] The biodiversity of marine ecosystems is diverse and a high number of species coexist side by side. However, despite the fact that most of these species share a common fertilization strategy, a high variability in terms of the size, shape, and motion of spermatozoa can be found. In this study, we have analyzed both the sperm motion parameters and the spermatozoa morphometric features of two swimmer (pufferfish and European eel) and two sessile (sea urchin and ascidian) marine species. The most important differences in the sperm motion parameters were registered in the swimming period. Sessile species sperm displayed notably higher values than swimmer species sperm. In addition, the sperm motilities and velocities of the swimmer species decreased sharply once the sperm was activated, whereas the sessile species were able to maintain their initial values for a long time. These results are linked directly to the species-specific lifestyles. Although sessile organisms, which show limited or no movement, need sperm with a capacity to swim for long distances to find the oocytes, swimmer organisms can move toward the female and release gametes near it, and therefore the spermatozoa does not need to swim for such a long time. At the same time, sperm morphology is related to sperm motion parameters, and in this study an in-depth morphometric analysis of ascidian, sea urchin, and pufferfish spermatozoa, using computer-assisted sperm analysis software, has been carried out for the first time. A huge variability in shapes, sizes, and structures of the studied species was found using electron microscopy. (C) 2014 Elsevier Inc. All rights reserved.This study was funded by the Spanish Ministry of Science and Innovation (MICINN; AGL2010-16009). Victor Gallego has a predoctoral grant (MICINN; BES-2009-020310) and has been granted a fellowship (EEBB-I-12-05858) of the Spanish Personnel Research Training Program to carry out this research in the Misaki Marine Biological Station (Miura, Japan).Gallego Albiach, V.; Pérez Igualada, LM.; Asturiano Nemesio, JF.; Yoshida, M. (2014). Sperm motility parameters and spermatozoa morphometric characterization in marine species: a study of swimmer and sessile species. Theriogenology. 82(5):668-676. https://doi.org/10.1016/j.theriogenology.2014.05.026S66867682

Differential expression of genes mapping to recurrently abnormal chromosomal regions characterize neuroblastic tumours with distinct ploidy status

<p>Abstract</p> <p>Background</p> <p>Neuroblastic tumours (NBTs) represent a heterogeneous spectrum of neoplastic diseases associated with multiple genetic alterations. Structural and numerical chromosomal changes are frequent and are predictive parameters of NBTs outcome. We performed a comparative analysis of the biological entities constituted by NBTs with different ploidy status.</p> <p>Methods</p> <p>Gene expression profiling of 49 diagnostic primary NBTs with ploidy data was performed using oligonucleotide microarray. Further analyses using Quantitative Real-Time Polymerase Chain Reaction (Q-PCR); array-Comparative Genomic Hybridization (aCGH); and Fluorescent <it>in situ </it>Hybridization (FISH) were performed to investigate the correlation between aneuploidy, chromosomal changes and gene expression profiles.</p> <p>Results</p> <p>Gene expression profiling of 49 primary near-triploid and near-diploid/tetraploid NBTs revealed distinct expression profiles associated with each NBT subgroup. A statistically significant portion of genes mapped to 1p36 (<it>P </it>= 0.01) and 17p13-q21 (<it>P </it>< 0.0001), described as recurrently altered in NBTs. Over 90% of these genes showed higher expression in near-triploid NBTs and the majority are involved in cell differentiation pathways. Specific chromosomal abnormalities observed in NBTs, 1p loss, 17q and whole chromosome 17 gains, were reflected in the gene expression profiles. Comparison between gene copy number and expression levels suggests that differential expression might be only partly dependent on gene copy number. Intratumoural clonal heterogeneity was observed in all NBTs, with marked interclonal variability in near-diploid/tetraploid tumours.</p> <p>Conclusion</p> <p>NBTs with different cellular DNA content display distinct transcriptional profiles with a significant portion of differentially expressed genes mapping to specific chromosomal regions known to be associated with outcome. Furthermore, our results demonstrate that these specific genetic abnormalities are highly heterogeneous in all NBTs, and suggest that NBTs with different ploidy status may result from different mechanisms of aneuploidy driving tumourigenesis.</p

Differential expression of genes mapping to recurrently abnormal chromosomal regions characterize neuroblastic tumours with distinct ploidy status

<p>Abstract</p> <p>Background</p> <p>Neuroblastic tumours (NBTs) represent a heterogeneous spectrum of neoplastic diseases associated with multiple genetic alterations. Structural and numerical chromosomal changes are frequent and are predictive parameters of NBTs outcome. We performed a comparative analysis of the biological entities constituted by NBTs with different ploidy status.</p> <p>Methods</p> <p>Gene expression profiling of 49 diagnostic primary NBTs with ploidy data was performed using oligonucleotide microarray. Further analyses using Quantitative Real-Time Polymerase Chain Reaction (Q-PCR); array-Comparative Genomic Hybridization (aCGH); and Fluorescent <it>in situ </it>Hybridization (FISH) were performed to investigate the correlation between aneuploidy, chromosomal changes and gene expression profiles.</p> <p>Results</p> <p>Gene expression profiling of 49 primary near-triploid and near-diploid/tetraploid NBTs revealed distinct expression profiles associated with each NBT subgroup. A statistically significant portion of genes mapped to 1p36 (<it>P </it>= 0.01) and 17p13-q21 (<it>P </it>< 0.0001), described as recurrently altered in NBTs. Over 90% of these genes showed higher expression in near-triploid NBTs and the majority are involved in cell differentiation pathways. Specific chromosomal abnormalities observed in NBTs, 1p loss, 17q and whole chromosome 17 gains, were reflected in the gene expression profiles. Comparison between gene copy number and expression levels suggests that differential expression might be only partly dependent on gene copy number. Intratumoural clonal heterogeneity was observed in all NBTs, with marked interclonal variability in near-diploid/tetraploid tumours.</p> <p>Conclusion</p> <p>NBTs with different cellular DNA content display distinct transcriptional profiles with a significant portion of differentially expressed genes mapping to specific chromosomal regions known to be associated with outcome. Furthermore, our results demonstrate that these specific genetic abnormalities are highly heterogeneous in all NBTs, and suggest that NBTs with different ploidy status may result from different mechanisms of aneuploidy driving tumourigenesis.</p