Instituto Valenciano Of Investigaciones Agrarias Cita Ivia

Segorbe, Spain

Instituto Valenciano Of Investigaciones Agrarias Cita Ivia

Segorbe, Spain
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Salvador I.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Cebrian-Serrano A.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Salamone D.,University of Buenos Aires | Silvestre M.A.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia
Spanish Journal of Agricultural Research | Year: 2011

The aim of this study was to identify the in vitro development stage at which the culture of a single or low number (n = 5 or 10) of oocytes/embryos could impair development in comparison with culture in group (n = 50). In the Experiment 1, it was confirmed that single in vitro embryo production yielded lower cleavage and blastocyst rates than in group (49.4 vs. 83.0%; 0% vs. 37.8%, respectively; p < 0.05). In Experiment 2 and 3, it was observed no effect on embryo development of culturing single or low number of oocytes during maturation and fertilization, respectively. In Experiment 4, it was observed a detrimental effect on blastocyst rate when cultured single or low number of embryos during post-fertilization in vitro culture (2.9; 10.2-10.8; 33.2% in single, low number of embryos (5-10), and controlgrouped, respectively; p < 0.05). In Experiment 5, it was observed that the last part of the culture period (day 3 onwards) seemed to be more affected by the low number of embryos placed in culture. In conclusion, post-fertilization culture, especially on days 3 to 7 after fertilization, seems to be the most important stage for embryo development on single and/or low number (5-10) of embryos culture.


Guinane C.M.,Roslin Institute | Guinane C.M.,Teagasc | Penades J.R.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Fitzgerald J.R.,Roslin Institute
Virulence | Year: 2011

Staphylococcus aureus is an important human pathogen that also causes economically important infections of livestock. In a recent paper, we employed a population genomic approach to investigate the molecular basis of ruminant host adaptation by S. aureus. The data suggest that the common pathogenic clone associated with small ruminants originated in humans but has since adapted to its adopted host through a combination of allelic diversification, gene loss and acquisition of mobile genetic elements. In particular, a new subfamily of staphylococcal pathogenicity islands (SaPI) was identified encoding a novel von Willebrand factor-binding protein (vWBP) with ruminant-specific coagu-lase activity. The wide distribution of vWBP-encoding SaPIs among ruminant strains implies an important role in host-adaptation. In the current article we summarize the findings of the paper and comment on the implications of the study for our understanding of the molecular basis of bacterial host adaptation. © 2011 Landes Bioscience.


Marti M.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Trotonda M.P.,Public University of Navarra | Tormo-Mas M.A.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Vergara-Irigaray M.,Public University of Navarra | And 4 more authors.
Microbes and Infection | Year: 2010

In Staphylococcus aureus, biofilm formation can be associated with the production of surface-anchored proteins, including Bap, SasG, FnBPs or Spa. By mutational analysis, and using a model strain in which biofilm formation was Bap-dependent, we found that σB was essential for protein-dependent biofilm development. Non-polar mutations of σB in genetically unrelated S. aureus strains lowered the Bap expression and completely impaired biofilm development. Although Northern blot analysis showed a slight reduction in bap transcription, we demonstrated that Aur and SspA, two proteases that are overexpressed in the sigB mutant strain and are capable of degrading Bap, inhibit biofilm formation. Interestingly, a double sigB-agr mutant, which showed a diminished capacity to express extracellular proteases, was able to restore biofilm formation. Since the vast majority of the S. aureus global regulators control the expression of the extracellular proteases, the results of this work demonstrate the existence of a new pathway controlling protein-mediated biofilm formation in which different global regulators modulate biofilm formation by controlling the expression of S. aureus extracellular proteases. © 2009 Elsevier Masson SAS. All rights reserved.


Ubeda C.,New York University | Ubeda C.,Center for Advanced Research in Public Health | Tormo-Mas M.T.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Penades J.R.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Novick R.P.,New York University
Plasmid | Year: 2012

The SaPIs and their relatives are phage satellites and are unique among the known bacterial pathogenicity islands in their ability to replicate autonomously. They possess a phage-like replicon, which is organized as two sets of iterons arrayed symmetrically to flank an AT-rich region that is driven to melt by the binding of a SaPI-specific initiator (Rep) to the flanking iterons. Extensive deletion analysis has revealed that Rep can bind to a single iteron, generating a simple shift in a gel mobility assay; when bound on both sides, a second retarded band is seen, suggesting independent binding. Binding to both sites of the ori is necessary but not sufficient to melt the AT-rich region and initiate replication. For these processes, virtually the entire origin must be present. Since SaPI replication can be initiated on linear DNA, it is suggested that bilateral binding may be necessary to constrain the intervening DNA to enable Rep-driven melting. © 2012 Elsevier Inc..


Viana D.,CEU Cardenal Herrera University | Comos M.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | McAdam P.R.,Roslin Institute | Ward M.J.,University of Edinburgh | And 8 more authors.
Nature Genetics | Year: 2015

The capacity of microbial pathogens to alter their host tropism leading to epidemics in distinct host species populations is a global public and veterinary health concern. To investigate the molecular basis of a bacterial host-switching event in a tractable host species, we traced the evolutionary trajectory of the common rabbit clone of Staphylococcus aureus. We report that it evolved through a likely human-to-rabbit host jump over 40 years ago and that only a single naturally occurring nucleotide mutation was required and sufficient to convert a human-specific S. aureus strain into one that could infect rabbits. Related mutations were identified at the same locus in other rabbit strains of distinct clonal origin, consistent with convergent evolution. This first report of a single mutation that was sufficient to alter the host tropism of a microorganism during its evolution highlights the capacity of some pathogens to readily expand into new host species populations. © 2015 Nature America, Inc. All rights reserved.


Blanch E.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Tomas C.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Graham J.K.,Colorado State University | Moce E.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia
Reproduction in Domestic Animals | Year: 2012

Contents: Cryopreserved boar sperm is not used extensively for artificial insemination, owing to the poor fertility rates of the sperm after freezing and thawing. The sperm membrane is damaged as the cells are cooled from body temperature to 5°C (cold shock), as well as during the freeze-thaw process. Increasing the cholesterol content of boar sperm membranes could help them survive cryopreservation, similar to sperm from other species that are cold shock sensitive. The aim of this study was to determine the optimal cholesterol-loaded cyclodextrin (CLC) concentration to use for boar sperm cryopreservation, and the influence of CLCs on the cryosurvival of sperm from boars classified as good or poor freezers. Treating boar sperm with 1mg of CLC/120×106 sperm slightly improved (p<0.05) the percentage of viable sperm after freezing-thawing. On the other hand, sperm, from both good and poor freezers, responded similarly to CLC treatment. Nevertheless, additional studies will be needed to study the effect of this treatment on other parameters of sperm quality. © 2012 Blackwell Verlag GmbH.


Quiles-Puchalt N.,Institute Biomedicina Of Valencia Ibv Csic | Quiles-Puchalt N.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Tormo-Mas M.A.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Tormo-Mas M.A.,CEU Cardenal Herrera University | And 9 more authors.
Nucleic Acids Research | Year: 2013

The propagation of bacteriophages and other mobile genetic elements requires exploitation of the phage mechanisms involved in virion assembly and DNA packaging. Here, we identified and characterized four different families of phage-encoded proteins that function as activators required for transcription of the late operons (morphogenetic and lysis genes) in a large group of phages infecting Gram-positive bacteria. These regulators constitute a super-family of proteins, here named late transcriptional regulators (Ltr), which share common structural, biochemical and functional characteristics and are unique to this group of phages. They are all small basic proteins, encoded by genes present at the end of the early gene cluster in their respective phage genomes and expressed under cI repressor control. To control expression of the late operon, the Ltr proteins bind to a DNA repeat region situated upstream of the terS gene, activating its transcription. This involves the C-terminal part of the Ltr proteins, which control specificity for the DNA repeat region. Finally, we show that the Ltr proteins are the only phage-encoded proteins required for the activation of the packaging and lysis modules. In summary, we provide evidence that phage packaging and lysis is a conserved mechanism in Siphoviridae infecting a wide variety of Gram-positive bacteria. © 2013 The Author(s).


Mir-Sanchis I.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Martinez-Rubio R.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Martinez-Rubio R.,CEU Cardenal Herrera University | Marti M.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | And 7 more authors.
Molecular Microbiology | Year: 2012

Staphylococcus aureus pathogenicity islands (SaPIs) are a group of related 15-17kb mobile genetic elements that commonly carry genes for superantigen toxins and other virulence factors. The key feature of their mobility is the induction of SaPI excision and replication by certain phages and their efficient encapsidation into specific small-headed phage-like infectious particles. Previous work demonstrated that chromosomal integration depends on the SaPI-encoded recombinase, Int. However, although involved in the process, Int alone was not sufficient to mediate efficient SaPI excision from chromosomal sites, and we expected that SaPI excision would involve an Xis function, which could be encoded by a helper phage or by the SaPI, itself. Here we report that the latter is the case. In vivo recombination assays with plasmids in Escherichia coli demonstrate that SaPI-coded Xis is absolutely required for recombination between the SaPI attL and attR sites, and that both sites, as well as their flanking SaPI sequences, are required for SaPI excision. Mutational analysis reveals that Xis is essential for efficient horizontal SaPI transfer to a recipient strain. Finally, we show that the master regulator of the SaPI life cycle, Stl, blocks expression of int and xis by binding to inverted repeats present in the promoter region, thus controlling SaPI excision. © 2012 Blackwell Publishing Ltd.


Penades J.R.,Institute Biomedicina Of Valencia | Penades J.R.,CEU Cardenal Herrera University | Donderis J.,Institute Biomedicina Of Valencia | Donderis J.,Research Center Biomedica En Red Of Enfermedades Raras Ciberer | And 4 more authors.
Current Opinion in Microbiology | Year: 2013

Deciphering the molecular mechanisms that control relevant cellular processes is of utmost importance to understand how viruses, prokaryotic and eukaryotic cells work. The diversity of living organisms suggests that there are novel regulators still to be discovered, which may uncover new regulatory paradigms. dUTPases (Duts) are assumed to be ubiquitous enzymes regulating cellular dUTP levels to prevent misincorporation of uracil into DNA. Recently however, Duts have been involved in the control of several relevant cellular processes, including transfer of mobile genetic elements, regulation of the immune system, autoimmunity or apoptosis, suggesting that they perform regulatory functions. This review aims at investigating the unexplored impact of Duts as novel signalling molecules. © 2013 Elsevier Ltd.


Tormo-Mas M.T.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Donderis J.,Institute Biomedicina Of Valencia | Garcia-Caballer M.,Instituto Valenciano Of Investigaciones Agrarias Cita Ivia | Garcia-Caballer M.,CEU Cardenal Herrera University | And 6 more authors.
Molecular Cell | Year: 2013

dUTPases (Duts) have emerged as promising regulatory molecules controlling relevant cellular processes. However, the mechanism underlying this regulatory function remains enigmatic. Using staphylococcal pathogenicity island (SaPI) repression as a model, we report here that phage Duts induce the transfer of SaPI-encoded virulence factors by switching between active (dUTP-bound) and inactive (apo state) conformations, a conversion catalyzed by their intrinsic dUTPase activity. Crystallographic and mutagenic analyses demonstrate that binding to dUTP reorders the C-terminal motif V of the phage-encoded Duts, rendering these proteins into the active conformation required for SaPI derepression. By contrast, the conversion to the apo state conformation by hydrolysis of the bound dUTP generates a protein that is unable to induce the SaPI cycle. Because none of the requirements involving Duts in SaPI transfer are exclusive to the phage-encoded proteins, we propose that Duts are widespread cellular regulators acting in a manner analogous to the eukaryotic G proteins. © 2013 Elsevier Inc.

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