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Munoz Mendoza M.,Servicio de Sanidad Animal | Juan L.D.,Complutense University of Madrid | Ocampo A.,Area Veterinaria de Lalin | Mourelo J.,Servicio de Sanidad Animal | And 5 more authors.
Veterinary Journal | Year: 2012

Tuberculosis was diagnosed in three flocks of sheep in Galicia, Spain, in 2009 and 2010. Two flocks were infected with Mycobacterium bovis and one flock was infected with Mycobacterium caprae. Infection was confirmed by the comparative intradermal tuberculin test, bacteriology, molecular analysis and histopathology. Sheep have the potential to act as a reservoir for tuberculosis. © 2011 Elsevier Ltd.

Trigal B.,SERIDA | Munoz M.,SERIDA | Gomez E.,SERIDA | Caamano J.N.,SERIDA | And 4 more authors.
Reproduction in Domestic Animals | Year: 2013

This work analyses the effects of a high hydrostatic pressure (HHP) treatment on in vitro survival of in vitro produced (IVP) bovine embryos vitrified with the Cryologic Vitrification Method (CVM). Consequences on embryo quality in terms of cell proliferation and differentiation, and levels of embryonic Heat Shock Protein 70 (Hsp-70) were also examined. Day 7 and 8 bovine in vitro-produced blastocysts were submitted to an HHP treatment (60MPa, at 32°C for 1h) and allowed to recover for 1 or 2h in culture medium. The HHP treatment did not improve blastocyst survival rates after vitrification/warming. Survival (24h post-warming) and hatching (48h post-warming) rates were 79.3±4.9 and 51.8±4.2 vs 73.9±4.2 and 44.7±4.1 for untreated controls and HHP-treated embryos, respectively. Total cell numbers measured in fresh embryos were reduced after 1h at 32°C, with or without HHP treatment, indicating that cell proliferation was stopped as a result of stress. Vitrified HHP-treated embryos that hatched at 48h after warming showed increased cell numbers in their ICM compared with untreated controls (50.2±3.1 vs 38.8±2.7), indicating higher embryo quality. Treatment of blastocysts with HHP did not alter the level of the Hsp-70 protein. In our conditions, HHP treatment did not affect the cryoresistance of these embryos. However, combination of HHP treatment and vitrification in fibreplugs resulted in an increase in the ICM cell number of hatched embryos 48h post-warming. © 2012 Blackwell Verlag GmbH.

Meijon M.,University of Oviedo | Meijon M.,Institute Biotecnologia Of Asturias | Feito I.,SERIDA | Valledor L.,University of Oviedo | And 5 more authors.
BMC Plant Biology | Year: 2010

Background: The ability to control the timing of flowering is a key strategy for planning production in ornamental species such as azalea, however it requires a thorough understanding of floral transition. Floral transition is achieved through a complex genetic network and regulated by multiple environmental and endogenous cues. Dynamic changes between chromatin states facilitating or inhibiting DNA transcription regulate the expression of floral induction pathways in response to environmental and developmental signals. DNA methylation and histone modifications are involved in controlling the functional state of chromatin and gene expression.Results: The results of this work indicate that epigenetic mechanisms such as DNA methylation and histone H4 acetylation have opposite and particular dynamics during the transition from vegetative to reproductive development in the apical shoots of azalea. Global levels of DNA methylation and histone H4 acetylation as well as immunodetection of 5-mdC and acetylated H4, in addition to a morphological study have permitted the delimitation of four basic phases in the development of the azalea bud and allowed the identification of a stage of epigenetic reprogramming which showed a sharp decrease of whole DNA methylation similar to that is defined in other developmental processes in plants and in mammals.Conclusion: The epigenetic control and reorganization of chromatin seem to be decisive for coordinating floral development in azalea. DNA methylation and H4 deacetylation act simultaneously and co-ordinately, restructuring the chromatin and regulating the gene expression during soot apical meristem development and floral differentiation. © 2010 Meijón et al; licensee BioMed Central Ltd.

Sanchez-Gomez D.,Technical University of Madrid | Sanchez-Gomez D.,Sustainable Forest Management Research Institute UVa INIA | Majada J.,SERIDA | Alia R.,SERIDA | And 4 more authors.
Annals of Forest Science | Year: 2010

Interpopulation variation in key functional traits of Pinus pinaster Ait. is well recognized. However, the relative importance of drought tolerance to explain this regional variation in the species remains elusive. Here, we raise the question whether water availability constitutes a likely driver of regional variation in biomass allocation, growth and morphological traits of ten populations that cover the distribution range of P. pinaster. We carried out an experiment where seedlings of five families per population were submitted to two contrasting watering treatments. The effects of water availability and population were significant for relative diameter and height growth rate, biomass allocation and number of lateral stems and dwarf shoots. Total dry mass significantly differed between watering treatments but it did not among populations. Populations could be clustered into four main groups. Root mass fraction explained most of the variation and significantly correlated to altitude but not to aridity.The geographical pattern of genetic variation found in morphology and biomass allocation did not translate into population differences in drought tolerance or phenotypic plasticity to water availability, indicating that water availability is not a likely driver of the regional variation observed in the studied traits of P. pinaster at the seedling stage. © 2010 INRA, EDP Sciences.

Aranda I.,Instituto Nacional Of Investigacion Y Tecnologia Agraria | Alia R.,Instituto Nacional Of Investigacion Y Tecnologia Agraria | Ortega U.,SERIDA | Dantas A.K.,SERIDA | Majada J.,SERIDA
Tree Genetics and Genomes | Year: 2010

The response to drought of 20 open-pollinated families from four Pinus pinaster Ait. populations covering a latitudinal cline (France, Central and Southern Spain, and Morocco) was assessed. The seedlings were cultivated in a greenhouse under controlled temperature and vapor pressure deficit for 120 days, and subjected to two watering regimes for 60 days. Different biomass partitioning variables, pre-dawn water potential, and isotopic discrimination of 13C in needles (Δ) as surrogate of long-term water use efficiency were estimated for each seedling at the end of the experiment. In response to the imposed drought, there was no change in the root biomass partitioning, but the overall Δ values decreased in response to water stress. All the families of the population from Morocco showed the highest investment in roots, regardless of the watering regime imposed. Inter-family differences within populations were also significant for most parameters as confirmed by the heritability values estimated (higher under the well-watered treatment). The studied P. pinaster populations showed different strategies of response to drought. This may represent an important mechanism by local populations in facing future climatic change. The results could be of value in forest conservation and breeding programs of maritime pine in the future. © Springer-Verlag 2009.

Sanchez-Gomez D.,Technical University of Madrid | Velasco-Conde T.,SERIDA | Cano-Martin F.J.,Technical University of Madrid | Angeles Guevara M.,Instituto Nacional Of Investigaciones Agrarias Inia | And 4 more authors.
Environmental and Experimental Botany | Year: 2011

Mediterranean trees are expected to be exposed to an increased aridity in the near future. The lack of genetic variability of stone pine (Pinus pinea L.) could limit the adaptation potential of the species to more severe droughts. However, information on intra-specific variability in adaptive traits that might confer potential drought tolerance is scant for stone pine. In this study, we explored the adaptation potential of stone pine to water deficit through variability of key functional traits. In particular, inter-clonal variation in whole plant performance in response to water deficit was studied. The role of inter-clonal variation in functional traits and phenotypic plasticity as determinants of whole plant performance under water deficit were also assessed. Relative growth rate (RGR) and key morphological and physiological traits were studied in ramets of 20 different clones of stone pine from a wide range of populations in the Iberian Peninsula. The plants were grown in climatic chamber under controlled environmental conditions and were submitted to an experimental water deficit of 20 days. The results showed significant inter-clonal variation in total dry mass, RGR, biomass partitioning specific leaf area (SLA), leaf area ratio (LAR), net photosynthetic rate (An), stomatal conductance (gs) and intrinsic water use efficiency (WUEi). Significant phenotypic plasticity in response to drought was found for An, gs and WUEi but it did not differ among clones. The clones could be clustered into different groups according to the variability of the studied traits in a principal components analysis. LAR, SLA and net photosynthetic rates explained most of the variation. A combination of traits (high LAR and low WUEi) agreeing with a water spending strategy rather than with a water saving strategy seemed to underlie enhanced RGR under water deficit. Stone pine showed high phenotypic plasticity and significant intra-specific variation in functional traits in response to water deficit. The pattern of variation found in morphological and physiological traits might allow adaptation to short term water deficit in this species. © 2010 Elsevier B.V.

Martinez-Alonso C.,CETEMAS | Kidelman A.,SERIDA | Feito I.,SERIDA | Velasco T.,SERIDA | And 3 more authors.
New Forests | Year: 2012

Due to the high economic importance of Pinus pinaster Ait., there is considerable interest in developing, improving and extending the use of its families for mass clonal propagation and in breeding programmes. In the current study, we evaluated shoot growth, rooting ability and mini-cuttings production of P. pinaster in response to nitrogen fertilization and seasons. We compared eight half-sib families of P. pinaster from Asturias and Galicia (Northern Iberian Peninsula), searching for useful parameters and growing conditions to be included in a mass propagation program for clonal family forestry. We fertilized P. pinaster seedling mother plants kept in a greenhouse with three levels of nitrogen: high (HN), medium (MN) and low (LN) to evaluate rooting ability of mini-cuttings. In addition, we evaluated the maximal potential production of rooted mini-cuttings considering nine cycles of propagation over 1 year, also using three levels of nitrogen. The HN treatment significantly influenced the rooting process, with length, area and volume of roots all being positively affected. Spring was the most favourable season for mini-cuttings in the HN treatment. This study provides valuable new information to optimize the clonal propagation protocol for P. pinaster and shows that the mini-cuttings technique has great potential in mass scale cloning, providing high quality sprout production and well-formed new plants. © 2012 Springer Science+Business Media B.V.

Ciordia M.,SERIDA | Feito I.,SERIDA | Pereira-Lorenzo S.,University of Santiago de Compostela | Fernandez A.,SERIDA | Majada J.,SERIDA
Environmental and Experimental Botany | Year: 2012

Variation patterns in phenotypic expression after cultivation in two water regimes: 50% and 90% substrate saturation were studied in progenies of chestnut (. Castanea sativa Mill.) cultivars originating from two main origins in the Iberian Peninsula: North (specifically from Asturias and Galicia) and Central Iberian Peninsula (specifically from the Canary Islands and Andalusia in Southern Spain related to this gene pool). The restricted water supply reduced the water potential of all the families as well as their growth, both in terms of height and weight. Root development increased whilst leaf area decreased as a result of reduction in number and surface area of individual leaves as well as sprouting rather than through defoliation. Drought stress also modified the morphology of the leaves and the relationships indicating the rate of relative development of the leaves compared to the rest of the plant. A correlation with the minimum summer rainfall in the regions of provenance was observed for several traits. The phenotypic variation observed was greater than expected from progenies of selected fruit cultivars. The results of this study show that (i) phenotypic variation exists between the two origins of variability corresponding to the two identified gene pools; (ii) the Northern group might be considered more tolerant to drought stress when considering plantlets' ability to recover water potential, though important phenotypic plasticity was found in both origins of variability; and (iii) the pattern of variation found in some physiological and morphological traits suggests that Spanish chestnut stands have great potential to respond to the expected increase in drought in the Iberian Peninsula over the present century. © 2011 Elsevier B.V..

Abendano N.,Tecnalia | Sevilla I.,Tecnalia | Prieto J.M.,Serida | Garrido J.M.,Tecnalia | And 2 more authors.
Journal of Clinical Microbiology | Year: 2012

Quantification of 11 clinical strains of Mycobacterium avium subsp. paratuberculosis isolated from domestic (cattle, sheep, and goat) and wildlife (fallow deer, deer, wild boar, and bison) animal species in an automatic liquid culture system (Bactec MGIT 960) was accomplished. The strains were previously isolated and typed using IS1311 PCR followed by restriction endonuclease analysis (PCR-REA) into type C, S, or B. A strain-specific quantification curve was generated for each M. avium subsp. paratuberculosis strain by relating the time to detection in the liquid culture system to the estimated log10 CFU in each inoculum. According to their growth curves, the tested M. avium subsp. paratuberculosis strains were classified into two distinct groups. The first group included the S-type strain isolated from goat and all the sheep strains with C, S, and B genotypes. A second group contained the C- and B-type strains isolated from cattle, goat, and wildlife animals with the exception of the fallow deer strain. The strains isolated from cattle or sheep showed similar strain-specific standard curves irrespective of their genotype. In contrast, the strains isolated from goat or from wildlife animal species varied in their rates of growth in liquid culture. Universal-standard curves and algorithms for the quantification of each group of strains were generated. In addition, the liquid culture system was compared with a real-time quantitative PCR system for the quantification of the 11 M. avium subsp. paratuberculosis strains. Correlations between the estimated log10 CFU and M. avium subsp. paratuberculosis DNA copy numbers were very high for all the tested strains (R ≥ 0.9). Copyright © 2012, American Society for Microbiology. All Rights Reserved.

Jordana J.,Autonomous University of Barcelona | Ferrando A.,Autonomous University of Barcelona | Marmi J.,Autonomous University of Barcelona | Avellanet R.,Autonomous University of Barcelona | And 2 more authors.
Livestock Science | Year: 2010

The Pallaresa cattle breed is a native population of beef cattle found in the western regions of the Catalan Pyrenees (Spain). Since the second half of the twentieth century, population size decreased dramatically in favour of the Bruna dels Pirineus breed. At the present time, it is classified as a relic breed on the brink of extinction by the FAO, as only 22 breeding animals remain alive (2 bulls and 20 cows). Its importance as genetic and cultural heritage and its contribution in maintaining the landscape and ecosystem, as well as its unquestionable interest as a tourist attraction, make it an irreplaceable element in this area. In this paper, we present the breed characterisation of the population at the molecular, genealogical and morphological levels, and the comparison and phylogenetic relationships with other local breeds, in order to contribute to the characterisation of this prominent genetic resource. Genealogical analyses showed that the Pallaresa population is highly inbred. The mean inbreeding for both the whole pedigree and the living population was 18.93% and 25.71%, respectively. A set of 15 microsatellites were typed on the available Pallaresa individuals to ascertain the genetic variability of the breed at the molecular level and to allow a direct comparison with a set of previously studied 19 local cattle breeds from Spain, Portugal and France. The average number of alleles per locus for the Pallaresa breed (4.3) was the lowest of the analysed breeds and the within-population genetic identity, assessed using molecular coancestry, the highest (0.437). The Pallaresa population showed high levels of differentiation from the other analysed breeds. The lowest kinship distance was assessed for the pairs formed with the Bruna dels Pirineus and Salers breeds (0.417 ± 0.010 and 0.427 ± 0.011, respectively). STRUCTURE analysis showed that most Pallaresa individuals were assigned to one distinct cluster, showing that the breed gathers a unique genetic background. Despite its dramatic population scenario, the Pallaresa breed still preserves moderate levels of genetic diversity. The results obtained from the current study can contribute to design appropriate conservation strategies to prevent its extinction in the near future. The implementation of a mating programme involving Pallaresa bulls and Bruna dels Pirineus cows selected according to genetic and morphological closeness to the Pallaresa breed is suggested to prevent the genetic impoverishment of the population. © 2010 Elsevier B.V.

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