Ciudad Real, Spain


Ciudad Real, Spain
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Pinheiro A.,University of Porto | De Mera I.G.F.,SaBio IREC CSIC UCLM JCCM | De Mera I.G.F.,Complutense University of Madrid | Alves P.C.,University of Porto | And 6 more authors.
Immunogenetics | Year: 2013

Among mammals, the European rabbit (Oryctolagus cuniculus) has a unique mechanism of generating the primary antibody repertoire. Despite having over 200 VH genes, the VH1 gene, the most d-proximal VH gene, is used in 80-90 % of VDJ rearrangements, while the remaining 10-20 % is encoded by the VHn genes that map at least 100 Kb upstream of VH1. The maintenance of the VHn genes usage in low frequency in VDJ rearrangements has been suggested to represent a relic of an ancestral immunologic response to pathogens. To address this question, we sequenced VDJ genes for another leporid, genus Lepus, which separated from European rabbit 12 million years ago. Approximately 25 VDJ gene sequences were obtained for each one of three Lepus europaeus individuals. We found that Lepus also uses the VHn genes in 5-10 % of its VDJ rearrangements. Our results show that the VHn genes are a conserved ancestral polymorphism that has been maintained in the leporids genome and is being used for the generation of VDJ rearrangements by both modern Lepus and Oryctolagus. © 2013 Springer-Verlag Berlin Heidelberg.

Pinheiro A.,University of Porto | Woof J.M.,University of Dundee | Almeida T.,University of Porto | Abrantes J.,University of Porto | And 5 more authors.
Open Biology | Year: 2014

Immunoglobulin G (IgG) is the predominant serum immunoglobulin and has the longest serum half-life of all the antibody classes. The European rabbit IgG has been of significant importance in immunological research, and is therefore well characterized. However, the IgG of other leporids has been disregarded. To evaluate the evolution of this gene in leporids, we sequenced the complete IGHG for six other genera: Bunolagus, Brachylagus, Lepus, Pentalagus, Romerolagus and Sylvilagus. The newly sequenced leporid IGHGgene has an organization and structure similar to that of the European rabbit IgG. A gradient in leporid IgG constant domain diversity was observed, with the CH1 being the most conserved and the CH3 the most variable domain. Positive selectionwas found to be acting on all constant domains, but with a greater incidence in the CH3 domain, where a cluster of three positively selected siteswas identified. In the hinge region, only three polymorphic positions were observed. The same hinge length was observed for all leporids. Unlike the variation observed for the European rabbit, all 11 Lepus species studied share exactly the same hinge motif, suggesting its maintenance as a result of an advantageous structure or conformation. © 2014 The Authors.

Queiros J.,SaBio IREC CSIC UCLM JCCM | Queiros J.,University of Porto | Vicente J.,SaBio IREC CSIC UCLM JCCM | Boadella M.,SaBio IREC CSIC UCLM JCCM | And 3 more authors.
Biological Journal of the Linnean Society | Year: 2014

The influences of management practices and past demographic history on genetic diversity are of critical relevance to sustainable practices and the conservation of wildlife populations. The red deer (Cervus elaphus) is an interesting model species to address these questions because it has a wide geographical distribution and it has been intensively managed for humans in the last decades. In the present study, we have analyzed the impact of recent management practices on the genetic diversity of Iberian red deer populations and assessed the genetic variation effects on population and individual fitness-related traits. Four populations subjected to distinct management systems were selected: Cabañeros (CB) and Doñana (DN), not hunted populations; Fraga/Caspe (FG/CP), open hunting area with very low or absent management; and PE, fenced private hunting estate founded 31 years ago through the introduction of deer of different origins. Ten microsatellites were amplified in a total of 172 individuals. Additionally, several fitness-related traits such as the presence of tuberculosis compatible lesions (TBCL), spleen weight (SW), and body length (BL) were estimated. We found a marked genetic variation and differentiation among populations, suggesting a strong population structure. In the fenced population, the introduction of genetically distinct animals has led to high genetic variability (no evidence of inbreeding) despite intensive management. Lower levels of genetic diversity were observed in two historically isolated natural populations (DN and FG/CP). The past demographic history of Iberian populations appears to be more relevant than the current management policy in shaping the genetic variability of natural populations. Population genetic diversity may correlate with life-history traits and disease susceptibility, which could compromise the conservation and management of these wildlife populations. Although no significant effects of individual genetic diversity (general and local effect hypotheses) were observed on TBCL, SW and BL, some single-locus effects had almost significant trends for the TBCL and SW traits. © 2013 The Linnean Society of London.

Richomme C.,Nancy laboratory for rabies and wildlife | Boadella M.,SaBio IREC CSIC UCLM JCCM | Courcoul A.,University Paris Est Creteil | Durand B.,University Paris Est Creteil | And 7 more authors.
PLoS ONE | Year: 2013

The Eurasian wild boar (Sus scrofa) is increasingly considered as a relevant actor in the epidemiology of animal tuberculosis (TB). Therefore, monitoring TB in wild boar becomes a key tool for establishing comprehensive control schemes for this disease. To estimate the exposure of free living wild boar to Mycobacterium tuberculosis complex (MTC) in France, a bovine-purified protein derivative based ELISA was used to test 2,080 archived serum samples of hunter-harvested animals in 58 French "départements". Two cut-off values were used for diagnostic interpretation: 0.2, recommended by the manufacturer (specificity: 96.43%; sensitivity: 72.6%), and 0.5 (specificity: 100%; sensitivity: 64%). During the same period, at the 0.2 cut-off, global true seroprevalence was 5.9% (IC95%: 4.3%-7.7%) and 76% of the sampled "départements" had seropositive wild boar, including seven cattle TB-free "départements. At the 0.5 cut-off, global true seroprevalence was 2.2% (IC95%: 1.5-3.2) and positive wild boar belonged to 21% of the "départements". All but one of these positive "départements" had reported at least one cattle TB outbreak since 2000. A good consistence between seropositive wild boar and TB outbreaks in cattle was found, especially at the 0.5 cut-off value (the mean distance to the nearest cattle TB outbreak was 13km and 27km for seropositive and seronegative wild boar, respectively; P<0.05). The use of an ELISA to detect MTC antibodies in wild boar has permitted the description of the geographic distribution of MTC contact in wild boar in France. Our results suggest that the ELISA could be used as a first screening tool to conduct TB surveillance in wild boar at a population level. High-risk wild boar populations (e.g. overabundant) could be tested and if identified positive by ELISA they should be surveyed in detail by combining pathology and culture. © 2013 Richomme et al.

Zanet S.,University of Turin | Trisciuoglio A.,University of Turin | Bottero E.,University of Turin | De Mera I.G.F.,SaBio IREC CSIC UCLM JCCM | And 3 more authors.
Parasites and Vectors | Year: 2014

Background: Piroplasmosis are among the most relevant diseases of domestic animals. Babesia is emerging as cause of tick-borne zoonosis worldwide and free-living animals are reservoir hosts of several zoonotic Babesia species. We investigated the epidemiology of Babesia spp. and Theileria spp. in wild ungulates and carnivores from Northern Italy to determine which of these apicomplexan species circulate in wildlife and their prevalence of infection. Methods. PCR amplification of the V4 hyper-variable region of the 18S rDNA of Babesia sp./Theileria sp was carried out on spleen samples of 1036 wild animals: Roe deer Capreolus capreolus (n = 462), Red deer Cervus elaphus (n = 52), Alpine Chamois Rupicapra rupicapra (n = 36), Fallow deer Dama dama (n = 17), Wild boar Sus scrofa (n = 257), Red fox Vulpes vulpes (n = 205) and Wolf Canis lupus (n = 7). Selected positive samples were sequenced to determine the species of amplified Babesia/Theileria DNA. Results: Babesia/Theileria DNA was found with a mean prevalence of 9.94% (IC95% 8.27-11.91). The only piroplasms found in carnivores was Theileria annae, which was detected in two foxes (0.98%; IC95% 0.27-3.49). Red deer showed the highest prevalence of infection (44.23%; IC95% 31.6-57.66), followed by Alpine chamois (22.22%; IC95% 11.71-38.08), Roe deer (12.55%; IC95% 9.84-15.89), and Wild boar (4.67%; IC95% 2.69-7.98). Genetic analysis identified Babesia capreoli as the most prevalent piroplasmid found in Alpine chamois, Roe deer and Red deer, followed by Babesia bigemina (found in Roe deer, Red deer and Wild boar), and the zoonotic Babesia venatorum (formerly Babesia sp. EU1) isolated from 2 Roe deer. Piroplasmids of the genus Theileria were identified in Wild boar and Red deer. Conclusions: The present study offers novel insights into the role of wildlife in Babesia/Theileria epidemiology, as well as relevant information on genetic variability of piroplasmids infecting wild ungulates and carnivores. © 2014 Zanet et al.; licensee BioMed Central Ltd.

PubMed | Cabarceno Park, SaBio IREC CSIC UCLM JCCM and University of León
Type: Journal Article | Journal: Reproduction in domestic animals = Zuchthygiene | Year: 2016

Sedimentation of spermatozoa occurs during long-term liquid storage and this may produce deleterious changes. Our aim was to apply gelatine supplementation during long-term pre-freezing storage of bear sperm, applying final dilution and 6% glycerol at room temperature and cool in straws. We tested four models of sperm storage using a 1:1 dilution in TTF-ULE-Bear extender (TesT-fructose-egg yolk-glycerol 6%): (i) second 1:1 dilution at room temperature (RT), cooling at 5C in a tube and final dilution (10010(6) spermml(-1) ) (Standard); (ii) final dilution at RT and cooling in a tube (FD-Tube); (iii) final dilution at RT and cooling in 0.25ml plastic straw (FD-Straw); and (iv) final dilution at RT in extender supplemented with 1.5% gelatine (Gelatine) and cooling in a 0.25ml plastic straw. A Standard sample was stored at 5C for 1hr (Control); the rest of the samples (Standard, FD-Tube, FD-Straw, Gelatine) were stored for 24 or 48hrs before freezing (10010(6) spermml(-1) , glycerol 6%). The quality of the samples was assessed for motility by CASA, and viability (SYBR-14/propidium iodide-PI-; VIAB), acrosomal status (PNA-FITC/PI; iACR) and apoptotic status (YO-PRO-1/PI; YOPRO-) by flow cytometry. At pre-freezing, after 48hr, Gelatine showed significantly higher viability (for VIAB and YOPRO-) and progressiveness (PM, LIN and STR). At 48hr, Gelatine showed similar YOPRO-, iACR, LIN, STR and ALH respect to Control. At both 24 and 48h post-thawing, Gelatine sample had similar scores for YOPRO-, iACR, LIN, STR, WOB and VIAB (only 24hr) when compared with Control, and lower for TM, PM, rapidPM, VAP and ALH. No differences were found among others experimental groups with respect to Control. In conclusion, gelatine could be a suitable alternative to preserve the viability and progressive motility of brown bear ejaculates during long-term pre-freezing storage at 5C.

Reusken C.B.E.M.,National Health Research Institute | Haagmans B.L.,Erasmus Medical Center | Muller M.A.,University of Bonn | Gutierrez C.,University of Las Palmas de Gran Canaria | And 25 more authors.
The Lancet Infectious Diseases | Year: 2013

Background: A new betacoronavirus-Middle East respiratory syndrome coronavirus (MERS-CoV)-has been identified in patients with severe acute respiratory infection. Although related viruses infect bats, molecular clock analyses have been unable to identify direct ancestors of MERS-CoV. Anecdotal exposure histories suggest that patients had been in contact with dromedary camels or goats. We investigated possible animal reservoirs of MERS-CoV by assessing specific serum antibodies in livestock. Methods: We took sera from animals in the Middle East (Oman) and from elsewhere (Spain, Netherlands, Chile). Cattle (n=80), sheep (n=40), goats (n=40), dromedary camels (n=155), and various other camelid species (n=34) were tested for specific serum IgG by protein microarray using the receptor-binding S1 subunits of spike proteins of MERS-CoV, severe acute respiratory syndrome coronavirus, and human coronavirus OC43. Results were confirmed by virus neutralisation tests for MERS-CoV and bovine coronavirus. Findings: 50 of 50 (100%) sera from Omani camels and 15 of 105 (14%) from Spanish camels had protein-specific antibodies against MERS-CoV spike. Sera from European sheep, goats, cattle, and other camelids had no such antibodies. MERS-CoV neutralising antibody titres varied between 1/320 and 1/2560 for the Omani camel sera and between 1/20 and 1/320 for the Spanish camel sera. There was no evidence for cross-neutralisation by bovine coronavirus antibodies. Interpretation: MERS-CoV or a related virus has infected camel populations. Both titres and seroprevalences in sera from different locations in Oman suggest widespread infection. Funding: European Union, European Centre For Disease Prevention and Control, Deutsche Forschungsgemeinschaft. © 2013 Elsevier Ltd.

Beerli O.,University of Bern | Blatter S.,University of Bern | Boadella M.,SaBio IREC CSIC UCLM JCCM | Schoning J.,University of Bern | And 2 more authors.
Veterinary Journal | Year: 2015

Bovine tuberculosis (bTB) is a (re-)emerging disease in European countries, including Switzerland. This study assesses the seroprevalence of infection with Mycobacterium bovis and closely related agents in wild boar (Sus scrofa) in Switzerland, because wild boar are potential maintenance hosts of these pathogens. The study employs harmonised laboratory methods to facilitate comparison with the situation in other countries. Eighteen out of 743 blood samples tested seropositive (2.4%, CI: 1.5-3.9%) by ELISA, and the results for 61 animals previously assessed using culture and PCR indicated that this serological test was not 100% specific for M.bovis, cross-reacting with M.microti. Nevertheless, serology appears to be an appropriate test methodology in the harmonisation of wild boar testing throughout Europe. In accordance with previous findings, the low seroprevalence found in wild boar suggests wildlife is an unlikely source of the M.bovis infections recently detected in cattle in Switzerland. This finding contrasts with the epidemiological situation pertaining in southern Spain. © 2014 Elsevier Ltd.

PubMed | University of Murcia, SaBio IREC CSIC UCLM JCCM and Medianilla S.L. Finca Las Lomas
Type: Journal Article | Journal: Reproduction in domestic animals = Zuchthygiene | Year: 2016

The aims of this study were to assess the effects of the sex-sorting process on post-thaw sperm quality as well as on induced oxidative stress damage (H2 O2 0mm=H000; H2 O2 50mm=H050; H2 O2 100mm=H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non-sorted (NS) red deer spermatozoa incubated at 37C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p<0.05) for NS (59%3.3) than BSS (36.9%5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p<0.05) for BSS (21.6%5.0) than NS sperm (14.6%1.2). The presence of H2 O2 increased DNA damage in NS (H000 =4.1%0.9; H050=9.3%0.7; and H100=10.9%2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p<0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.54.8 vs 21.13.9 and BSS: 33.38.1 vs 8.91.8). These results demonstrate that the sperm-sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non-sorted sperm. Moreover, addition of GSH at 1mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.

Anderson L.G.,University of York | Gortazar C.,SaBio IREC CSIC UCLM JCCM | Vicente J.,SaBio IREC CSIC UCLM JCCM | Hutchings M.R.,Roslin Institute | White P.C.L.,University of York
Wildlife Research | Year: 2013

Context Bovine tuberculosis is a persistent disease of livestock in many parts of the world, especially where wildlife hosts co-exist with livestock. In south-western Spain, despite the widespread implementation of test-and-cull strategies for cattle, the herd prevalence in areas with high wild boar densities remains stable. The control of M. bovis infection in wild boar is likely to be essential for effective disease control in livestock. Methods We developed an individual-based model to evaluate whether vaccinating wild boar piglets with oral bait would be an effective strategy to reduce the prevalence of M. bovis infection in wild boar populations. Specifically, we quantified the proportion of piglets requiring vaccination and the number of years the vaccination programme would need to continue to eradicate bTB from wild boar within 25 years, comparing 'managed' populations on hunting estates where supplementary food is provided, with 'unmanaged', free-living populations. Successful vaccination was defined as the proportion of piglets that were delivered the vaccine and were effectively protected from infection. Key results Longer-term (25-year) vaccination strategies were more successful than short-term (5-year) strategies at either eradicating M. bovis or reducing it to below 90% of its original prevalence. M. bovis infection could be eradicated under a 25-year vaccination strategy if 80% of piglets were vaccinated in a managed population or 70% of piglets were vaccinated in an unmanaged population. In contrast, 5-year strategies in which 80% of piglets were vaccinated reduced the prevalence of M. bovis only by 27% or 8% in the managed and unmanaged populations, respectively. Conclusions The results of our simulation model, coupled with the promising results of initial vaccine and oral bait-uptake trials in wild boar indicated that vaccination could be an effective strategy to reduce the prevalence of M. bovis infection in wild boar if used in conjunction with other disease-control measures. Implications The vaccination of piglets over a long-term period has the potential to make an important contribution to the eradication of M. bovis infection from wild boar reservoirs in southern Spain. © CSIRO 2013.

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