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Many people worldwide suffer from hepatitis C virus (HCV) infection, which is frequently persistent. The lack of efficient vaccines against HCV and the unavailability of or limited compliance with existing antiviral therapies is problematic for health care systems worldwide. Improved small animal models would support further hepacivirus research, including development of vaccines and novel antivirals. The recent discovery of several mammalian hepaciviruses may facilitate such research. In this study, we demonstrated that bank voles (Clethrionomys glareolus) were susceptible to bank vole-associated Hepacivirus F and Hepacivirus J strains, based on the detection of hepaciviral RNA in 52 of 55 experimentally inoculated voles. In contrast, interferon α/β receptor deficient C57/Bl6 mice were resistant to infection with both bank vole hepaciviruses (BvHVs). The highest viral genome loads in infected voles were detected in the liver, and viral RNA was visualized by in situ hybridization in hepatocytes, confirming a marked hepatotropism. Furthermore, liver lesions in infected voles resembled those of HCV infection in humans. In conclusion, infection with both BvHVs in their natural hosts shares striking similarities to HCV infection in humans and may represent promising small animal models for this important human disease.

Many people worldwide suffer from hepatitis C virus (HCV) infection, which is frequently persistent. The lack of efficient vaccines against HCV and the unavailability of or limited compliance with existing antiviral therapies is problematic for health care systems worldwide. Improved small animal models would support further hepacivirus research, including development of vaccines and novel antivirals. The recent discovery of several mammalian hepaciviruses may facilitate such research. In this study, we demonstrated that bank voles (Clethrionomys glareolus) were susceptible to bank vole-associated Hepacivirus F and Hepacivirus J strains, based on the detection of hepaciviral RNA in 52 of 55 experimentally inoculated voles. In contrast, interferon α/β receptor deficient C57/Bl6 mice were resistant to infection with both bank vole hepaciviruses (BvHVs). The highest viral genome loads in infected voles were detected in the liver, and viral RNA was visualized by in situ hybridization in hepatocytes, confirming a marked hepatotropism. Furthermore, liver lesions in infected voles resembled those of HCV infection in humans. In conclusion, infection with both BvHVs in their natural hosts shares striking similarities to HCV infection in humans and may represent promising small animal models for this important human disease.

Wildlife management and conservation requires monitoring of species distribution and population indicators, especially when the unbalanced demographic changes of some species can affect the whole ecosystem functioning. The populations of wild boar (Sus scrofa) have, over the past few decades, undergone an expansion around the world, reaching situations of overabundance that can cause serious economic, ecological and health problems. This numerical increase of wild boar and its new spatial invasion can affect certain vulnerable species in sensitive ecological zones, such as the main inland wetland complex in SW Europe. In this context, we aim to (i) examine the association of wild boar abundance and that of lagomorphs, and waterbird productivity, controlling the possible effects of wild carnivores and other environmental predictors, and (ii) applying a survey method useful for managers to evaluate expected effect of wild boar abundance. Overall, the presence of wild boar has been detected in 80.8% of the monitored wetlands (n = 26), but a high variation of abundance rates was found. Wild boar abundance negatively associated with the productivity of the entire community of waterbirds, a priori highly vulnerable, breeding on shores or islands, both colonially (genus Gelochelidon, Himantopus, Recurvirostra, Sternula, Glareola, Tringa and Phoenicopterus) or solitary (genus Anas, Spatula, Mareca, Charadrius, Vanellus, Anser, Ardea, Aythya, Fulica, Netta, Oxyura and Tadorna). In addition, we evidenced a clear negative association of wild boar and wild rabbit (Oryctolagus cuniculus) population abundance in the monitored wetlands. Rabbits are a keystone species in the study area and represent stable prey for many endangered predators. Our results suggest that potential impacts caused by high wild boar abundance may be already reaching unsustainable levels for some wetlands. Inter-species integrated monitoring is key to address the population management plans of wild boar populations in wetlands and to preserve the most vulnerable species., The present work has benefited from financial support of LIFE project “Humedales de la Mancha”. JAB and AJC were supported by a “Juan de la Cierva” contract (FJCI-2015-23643 and FJCI-2017-33114) from MINECO-UCM and MINECO-UCLM, respectively.

Animal tuberculosis (TB), which is caused by the Mycobacterium tuberculosis complex (MTC), is a zoonotic disease of global concern, and has a wide variety of wild and domestic reservoirs that can establish complex epidemiological systems. Of all the strategies employed to control TB, reducing the risks of interaction at the wildlife-livestock interface is a cornerstone. However, detailed protocols with which to assess and implement farm-specific preventive actions that can be employed against interactions with wildlife are lacking for extensive production systems. We describe an On-farm Wildlife Risk Mitigation Protocol that is applicable to beef cattle farming in Mediterranean environments in order to control the wildlife-livestock interaction and MTC transmission through the use of Farm-specific Action Plans (FsAP). We assessed the implementation and verification of FsAP in terms of its practical feasibility and acceptability by farmers (n=55 farms). Of the potential risk points, waterers (41.3 %) and waterholes (24.4 %) were the most common. Waterholes and springs were identified as the points with the greatest risks. Actions related to water management were essential on most farms (99 % of the high-risk points), as were those regarding wildlife management (36.4 % of the farms provided wild boar or cervids with supplementary food for hunting purposes). Overall, 75 % of the farmers adopted the plans to some extent, with an average of 31.8 % of actions implemented, but with high variability depending on the type of actions proposed. Farmers prioritised low-cost measures. Our results, in their entirety, indicate that the adoption of this On-farm Wildlife Risk Mitigation Protocol is practical and feasible in Mediterranean ecosystems, and can be easily transferred to professionals and adapted to other bioregions or epidemiological systems. The subsequent evaluation of FsAPs in terms of efficacy and cost-effectiveness, along with increasing their acceptance by farmers, are necessary steps for the further development of TB Risk Mitigation Programmes at a nationwide level., Funding was provided by projects AGL2013-48523-C3-1-R and AGL2016-76358-R (MINECO; AEI/FEDER, UE). This is also a contribution to the EU FEADER PDR projects “Alcudia” and “GOSTU” on farm biosafety. JMG holds a FPI pre-doctoral scholarship (BES-2015- 072206, MINECO). PA is supported by an extension of his “Ramón y Cajal” contract (RYC-2012-11970, MINECO-UCLM). SJR holds a PhD contract from the UCLM co-supported by the European Social Fund(2018/12504). JAB is supported by the European Union project (H2020 VACDIVA 862874).

Attaining and maintaining the Official Tuberculosis Free status continues to be a challenge when several domestic and wild hosts contribute to the maintenance of the Mycobacterium tuberculosis complex (MTC). Local tuberculosis hotspots are sometimes identified in cattle in low-prevalence regions. We have, therefore, studied one such hotspot in depth in order to produce an epidemiological diagnosis. Host population size and MTC prevalence were estimated in selected wildlife and in livestock, while on-cattle environmental DNA detection was additionally used as a proxy for risk of exposure at the farm (herd) level. Positive skin test reactors were found on16 of the 24 cattle farms studied in the period 2012–2016. Although all goats tested negative to the skin test during this period, MTC was confirmed in four sheep at slaughter, thus indicating an unknown prevalence of infection in this host species. With regard to wildlife, the prevalence of MTC infection based on culture was 8.8% in the case of wild boar (Sus scrofa), and the only road-killed badger (Meles meles) submitted for culture tested positive. Two criteria were employed to divide the cattle farms into higher or lower risk: tuberculosis testing results and environmental DNA detection. Environmental MTC DNA detection yielded significant differences regarding “use of regional pastures” and “proximity to woodland”. This study suggests that on-animal environmental DNA sampling may help when assessing contact risk as regards MTC in livestock at the herd level. This tool opens up new avenues of epidemiological research in complex multi-host settings., This work was supported by the Programa de Tecnologias Avanzadas en Vigilancia Sanitaria (TAVS) from the Comunidad de Madrid (ref. S2013/ABI-2747). JMG holds a FPI predoctoral scholarship (BES-2015- 072206), funded by MINECO. This is a contribution to MINECO grant CGL2017-89866 WildDriver and EU-FEDER. This is also a contribution to Valle de Alcudia pilot project SG-2019-02 from PDR-CLM, and to GOSTU project EU-FEADER (AEIAGRI-PNDR-MAPA, ref. 20190020007521).

The poultry red mite (PRM), Dermanyssus gallinae, is a hematophagous ectoparasite considered as the major pest in the egg-laying industry. Its pesticide-based control is only partially successful and requires the development of new control interventions such as vaccines. In this study, we follow a vaccinology approach to identify PRM candidate protective antigens. Based on proteomic data from fed and unfed nymph and adult mites, we selected a novel PRM protein, calumenin (Deg-CALU), which is tested as a vaccine candidate on an on-hen trial. Rhipicephalus microplus Subolesin (Rhm-SUB) was chosen as a positive control. Deg-CALU and Rhm-SUB reduced the mite oviposition by 35 and 44%, respectively. These results support Deg-CALU and Rhm-SUB as candidate protective antigens for the PRM control., This work was partially supported by the European Cooperation in Science and Technology (COST), Improving current understanding and research for sustainable control of the poultry red mite Dermanyssus gallinae – COREMI project (Action FA1404). JFLB was supported by Ministerio de Ciencia, Innovación y Universidades (Spain), Doctorado Industrial contract (DI-14-06917) and Sabiotec SA. MV was supported by the University of Castilla- La Mancha (Spain).

This article belongs to the Special Issue Farm Animal and Wildlife Zoonotic Microorganisms., The aim of this study was to evaluate the diagnostic performance of a multiplex bead assay for the simultaneous detection of antibodies against Mycobacterium bovis, Brucella suis, and Trichinella spiralis. Sera from Eurasian wild boar of known serological status for TB (64 seropositive, 106 seronegative), Brucella (30 seropositive, 39 seronegative), and Trichinella (21 seropositive, 97 seronegative) were used for the development and evaluation of the assay. Magnetic beads coated with recombinant MPB83 antigen (TB), a whole-cell B. suis 1330 antigen, and an E/S T. spiralis antigen were used for the detection of specific antibodies using Bio-Rad Bio-Plex technology. The sensitivities (Se) and specificities (Sp) of the multiplex assay were, for M. bovis, 0.98 and 0.86; for B. suis, 1.00 and 0.97; and for T. spiralis, 0.90 and 0.99 (Se and Sp, respectively). The results show the diagnostic potential of this assay for the simultaneous detection of antibodies against M. bovis, B. suis, and T. spiralis in wild boar., We thankfully acknowledge the financial support of the European Union Seventh Framework Programme (2007–2013) under grant agreement no. 222633 (WildTech) titled “Novel Technologies for Surveillance of Emerging and Re-emerging Infections of Wildlife”.

In 2012, a wild boar (Sus scrofa) tuberculosis (TB) control programme was set up in a wild boar farm by means of intramuscular (IM) vaccination with a heat-inactivated Mycobacterium bovis vaccine (IV). The goal was to assess safety and efficacy of the parenterally administered IV in a large farm setting with natural M. bovis circulation. Based on preceding results under laboratory conditions, we hypothesized that vaccinated piglets would show smaller scores of TB-compatible lesions (TBCL) than unvaccinated controls. After vaccination, no adverse reactions were detected by visual inspection or at post-mortem examination (n = 668 and 97, respectively). Post-mortem data on TBCL were available for 97 vaccinated wild boar and 182 controls. The observed TBCL prevalence was 4.1% (95% CI = 0.2–8%) and 12.1% (95% CI = 7.1–17.1%) for vaccinated and control wild boar, respectively (P 0.05). The results show that IV administered intramuscularly to wild boar piglets is safe and protects vaccinated individuals (66% reduction in TBCL prevalence) against natural challenge in a low-prevalence setting. In a context of increasing TB prevalence in wild boar in Mediterranean habitats, vaccination achieved a progressive though slow decline in lesion prevalence since the onset of the vaccination scheme. Hence, vaccination might contribute, along with other tools, to TB control in wild boar and in pigs., This is a contribution to Plan Nacional I+D+i AGL2014-56305 from MINECO and EU FEDER. IDD was supportedby a predoctoral grant from MINECO.

Mycobacterial diseases are important health issues in farmed deer. The single intradermal tuberculin test is the standard test for tuberculosis testing in deer. We studied two factors which might influence the response of deer to skin testing: the inoculation site and the injection device. Deer included in this study were 2.5 years old farmed red deer (Cervus elaphus) hinds (n = 80). Two areas of 3 cm × 3 cm were shaved at the left side of the neck. Site A (SA) was situated about 10 cm caudal to the head, while site B (SB) was 10 cm caudal to SA. All hinds received at the same time two 0.1 ml inoculations of Mycobacterium avium derived purified protein derivative (aPPD). One inoculation was made by syringe and the other one with the needle-free syringe Dermojet. To test the inoculation site effect, half of the animals were inoculated by Dermojet in SA and by syringe in SB to compare with the inoculation in SA by syringe and Dermojet in SB in the other half. No differences were recorded for the injection device nor for the inoculation site. Ten hinds had a skinfold increase larger than 30 tenths of mm by any injection device and inoculation site. Seven (9%) and 6 (8%) hinds were classified as positive by syringe and Dermojet, and at the anterior or posterior inoculation site, respectively. The distribution of skinfold thickness increases did not differ by injection device. Our findings support the needle-free Dermojet syringe as a suitable tool for skin-testing in red deer and suggest no relevant effect of the position of the inoculation site along the neck in red deer., This is a contribution to MINECO Plan Nacional and EU FEDER grant WILD DRIVER (CGL2017-89866). Julio Isla is beneficiary of a co-financed contract by European Social Fund (ESF) of European Union (EU) and Programa Operativo de Empleo Juvenil belonging to JCCM.

A. Che’ Amat et al., Animal tuberculosis (TB) caused by infection with Mycobacterium bovis and closely related members of the M. tuberculosis complex (MTC), is often reported in the Eurasian wild boar (. Sus scrofa). Tests detecting antibodies against MTC antigens are valuable tools for TB monitoring and control in suids. However, only limited knowledge exists on serology test performance in 2-6 month-old piglets. In this age-class, recent infections might cause lower antibody levels and lower test sensitivity. We examined 126 wild boar piglets from a TB-endemic site using 6 antibody detection tests in order to assess test performance. Bacterial culture (. n=. 53) yielded a M. bovis infection prevalence of 33.9%, while serum antibody prevalence estimated by different tests ranged from 19% to 38%, reaching sensitivities between 15.4% and 46.2% for plate ELISAs and between 61.5% and 69.2% for rapid immunochromatographic tests based on dual path platform (DPP) technology. The Cohen kappa coefficient of agreement between DPP WTB (Wildlife TB) assay and culture results was moderate (0.45) and all other serological tests used had poor to fair agreements. This survey revealed the ability of several tests for detecting serum antibodies against the MTC antigens in 2-6 month-old naturally infected wild boar piglets. The best performance was demonstrated for DPP tests. The results confirmed our initial hypothesis of a lower sensitivity of serology for detecting M. bovis-infected piglets, as compared to older wild boar. Certain tests, notably the rapid animal-side tests, can contribute to TB control strategies by enabling the setup of test and cull schemes or improving pre-movement testing. However, sub-optimal test performance in piglets as compared to that in older wild boar should be taken into account., This is a contribution to Spanish Government MINECO Plan Nacional I+D+I grant AGL2014-56305 and FEDER, to a contract between CDTI and Glenton, and to the EU FP7 grant WildTBvac #613779. Azlan Che Amat has a PhD grant from the Malaysian Government, and José Angel Barasona and Iratxe Diéz-Delgado acknowledge PhD grants from the Spanish Government.

[Aim]: Species distribution models based on occurrence data (SDM) are used to derive broad-scale estimates of relative abundance, but a triangular relationship between predicted suitability for species occurrence and abundance is commonly observed¿that is, a low suitability would correspond to low abundances, whereas a high suitability would correspond to either low or high abundances. To properly understand the factors behind this triangular relationship, we assessed: (1) the capacity of SDM to explain variations in population abundance, and (2) the effect of population growth in the dynamics on the triangular relationship by comparing its variation in two sampling periods in the case of a species that is increasing in numbers. [Location]: Mainland Spain. [Methods]: Data on roe deer (Capreolus capreolus) relative abundance were obtained for 441 localities, of which 145 were surveyed twice (2006–2007 and 2011–2013). Species distribution was modelled using the favourability function. The relationship between favourability values for species occurrence and abundances were explored using quantile regressions. [Results]: The number of roe deer is increasing on a regional scale. The results from the earlier survey showed a weaker relationship than those obtained from the survey carried out later. Growth rates were significantly related to the residuals of the quantile regression fitted to the earlier survey (the higher the residual difference between observed and expected abundances, the higher the population growth rates). [Main conclusions]: Our results support the use of the residuals of the quantile regression as a proxy of the population growth rate. They also partly support the interpretation of the upper limit of abundance delimited by the regression as the environmental carrying capacity. Overall, the results are of general interest when using SDM to predict the population abundance of expanding species, such as alien species and/or in changing environments., P. Acevedo was supported by the Spanish Ministerio de Economía y Competitividad (MINECO) and the Universidad de Castilla-La Mancha (UCLM) through a “Ramón y Cajal” contract (RYC-2012-11970). The present work has benefited from the financial aid of ESFA contract (ENETWILD-OC/EFSA/ALPHA/2016/01) and research grant by the Ministerio de Ciencia e Innovación (AGL2013-48523-C3-1-R).

The red deer (Cervus elaphus) is a widespread wild ungulate in Europe that has suffered strong anthropogenic impacts over their distribution during the last centuries, but also at the present time, due its economic importance as a game species. Here we focus on the evolutionary history of the red deer in Iberia, one of the three main southern refugial areas for temperate species in Europe, and addressed the hypothesis of a cryptic refugia at higher latitudes during the Last Glacial Maximum (LGM). A total of 911 individuals were sampled, genotyped for 34 microsatellites specifically developed for red deer and sequenced for a fragment of 670 bp of the mitochondrial (mtDNA) D-loop. The results were combined with published mtDNA sequences, and integrated with species distribution models and historical European paleo-distribution data, in order to further examine the alternative glacial refugial models and the influence of cryptic refugia on European postglacial colonization history. Clear genetic differentiation between Iberian and European contemporary populations was observed at nuclear and mtDNA levels, despite the mtDNA haplotypes central to the phylogenetic network are present across western Europe (including Iberia) suggesting a panmictic population in the past. Species distribution models, fossil records and genetic data support a timing of divergence between Iberian and European populations that overlap with the LGM. A notable population structure was also found within the Iberian Peninsula, although several populations displayed high levels of admixture as a consequence of recent red deer translocations. Five D-loop sub-lineages were found in Iberia that belong to the Western European mtDNA lineage, while there were four main clusters based on analysis of nuclear markers. Regarding glacial refugial models, our findings provide detailed support for the hypothesis that red deer may have persisted in cryptic northern refugia in western Europe during the LGM, most likely in southern France, southern Ireland, or in a region between them (continental shelf), and these regions were the source of individuals during the European re-colonization. This evidence heightens the importance of conserving the high mitochondrial and nuclear diversity currently observed in Iberian populations., This work was supported by: Portuguese national funds through the FCT (Fundação para a Ciência e a Tecnologia), FEDER funds (Fundo Europeu de Desenvolvimento Regional) through Programa Operacional Potencial Humano-Quadro de Referência Estratégico Nacional (POPH-QREN) from the European Social Fund and Portuguese Ministério da Educação e Ciência and the project ‘Genomics applied to genetic resources’, North Portugal Regional Operational Programme 2007/2013 (ON.2 – O Novo Norte); the EU grant ‘Harmonised approaches in monitoring wildlife population health, and ecology and abundance’ (APHAEA, 219235_FP7_ERA-NET_EMIDA) and CDTI. This is also a contribution to grant AGL20111-30041 from MINECO, Spain. MB is currently an employee and CEO of SABIOtec commercial company. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript and only provided financial support in the form of grants to the authors (SFRH/BD/73732/2010 PhD grant to JQ, SFRH/BD/5880/2008 PhD grant to JPVS and SFRH/BSAB/1278/2012 sabbatical grant to PCA) and/or research materials.

The Canary Islands contain the most important dromedary camel ( Camelus dromedarius) population in the European Union and are the main export point of dromedaries to continental Europe and Latin America. We investigated the presence of antibodies against relevant disease agents in 100 Canarian camel sera. Selected blood samples of the same animals were also tested by PCR. Sera were tested for antibodies against Bluetongue virus (BTV; 0%), Bovine Viral Diarrhoea virus (BVDV; 0%), Camelpox virus (CPV; 8% by serum neutralization, 16% by ELISA), Peste des Petits Ruminants virus (PPRV, 0%), Rift Valley Fever virus (RVFV; 0%) and West Nile Fever virus (WNV; 3%), the bacterial pathogens Anaplasma sp. (3%), Brucella sp. (1%), Coxiella burnetii (19%), Mycobacterium avium paratuberculosis (MAP; 22%), Mycobacterium tuberculosis complex (MTC; 10%) and Rickettsia sp. (83%), and the parasites Toxoplasma gondii (36%) and Neospora caninum (86%). The most remarkable findings were the detection of antibodies against CPV and the high antibody prevalence against C. burnetii, Rickettsia sp., T. gondii and N. caninum. By PCR, we found no C. burnetii, N. caninum and Anaplasma sp. DNA in the tested samples. However, Rickettsia sp. DNA was detected in six antibody positive tested samples. These results should be taken into consideration in order to implement adequate control measures and avoid a potential dissemination of infections to other territories., This is a contribution to EU grants ANTIGONE (278976) and APHAEA (EMIDA ERA-NET).

As a response to the need for improved and cost-efficient West Nile virus (WNV) and other flavivirus surveillance tools, we tested 887 juvenile free-living red deer, 742 free-living juvenile wild boar, and 327 farmed deer, to detect temporal variability in exposure to these viruses. Thirty of 742 wild boar samples (4%; 95% CI 2.8,5.7) yielded a positive ELISA result. Antibody-positive individuals had been sampled between 2003 and 2011 in localities from central and southern Spain. No wild boar from the northern half of Spain (n=120) tested positive. Regarding juvenile wild red deer, only two out of 887 samples yielded a positive ELISA result (0.2%; 95% CI 0.1,0.8). These two samples came from the same site and sampling year. The likelihood of detecting contact with WNV or cross-reacting flaviviruses was 18 times higher among juvenile wild boar than among juvenile red deer. ELISA positivity among farmed deer increased 10-fold after local flavivirus outbreaks recorded in the summer and autumn of 2010. This survey demonstrated the potential usefulness of juvenile wild ungulates, particularly wild boar, as suitable flavivirus sentinels in southwestern Europe, and that systematic serum banking of samples from hunter-harvested wildlife or from individual farmed ungulates provides valuable material for retrospective epidemiological surveys and future disease monitoring., This study was supported by POII09-0141-8176 (Castilla-La Mancha) and by JCCM PPIC10-0226-0243. The authors also acknowledge the support of the Ministerio de Medio Ambiente, Rural y Marino (MARM).

Wild boars are natural hosts for African swine fever (ASF). The ASF virus (ASFV) can persist for long periods in the environment, such as in ticks and contaminated products, which may be sources of infection for wild boar populations. African swine fever was eradicated in domestic pig populations in Spain in 1995, after 35 years of significant effort. To determine whether ASFV can persist in wild boar hosts after it has been eradicated from domestic pigs and to study the role of wild boar in helping ASFV persist in the environment, we checked for the presence of ASFV in wild boars in Donana National Park, one of the largest natural habitats of wild boar in Spain and one of the last areas where ASF was endemic prior its eradication. Samples from 158 animals collected between 2006 and 2010 were analysed using serological and nucleic acid-based diagnostic techniques recommended by the World Organization for Animal Health (OIE). None of the samples was found to be positive. These results confirm the absence of disease in wildlife in what was once one of the areas most affected by ASF in Spain, and they suggest that wild boars play a limited role in ASFV persistence. These results confirm that ASFV cannot persist in isolated wild boar populations for long periods of time without the interaction of other factors such as re-infection by contact with domestic pigs or by feeding on contaminated swill.

Summary A total of 1279 Eurasian wild boar (Sus scrofa) sera were collected from 2000 to 2011 in the Iberian Peninsula to reveal time changes in serum antibody prevalences against selected infectious agents (porcine circovirus type 2, PCV2; porcine reproductive and respiratory syndrome virus, PRRSV; hepatitis E virus, HEV; and Erysipelothrix rhusiopathiae) and to identify putative individual or population factors driving such changes. Overall seroprevalences were 48%, 26%, 2% and 15% for PCV2, HEV, PRRSV and E. rhusiopathiae, respectively. The global observed prevalence of antibodies against PCV2 and HEV remained stable during the study period, while the global mean antibody seroprevalence against E. rhusiopathiae declined. The mean increment in prevalence was always lower for open than for fenced sites. This study evidenced for the first time that wild boar from the Iberian Peninsula have widespread contact with E. rhusiopathiae, and confirmed high prevalences of antibodies against PCV2 and HEV. Maintained high prevalences of transmissible agents in wild boar suggest that epidemiological drivers such as aggregation and high density are still acting. This will most probably also affect the transmission rates of other disease agents and should be taken into account regarding disease control at the wildlife–livestock interface.

The current study is a contribution to Junta de Comunidades de Castilla-La Mancha (JCCM) PPIC10-0226-0243, Ministerio de Ciencia e Innovacion (MCINN) Plan Nacional I + D + i AGL2008-03875 and Fondo Europeo de Desarrollo Regional (FEDER) and to FP7 EU TB-STEP (212414). (Strategies for the eradication of bovine tuberculosis). Studies on diseases shared between domestic animals and wildlife are also supported by Santander and Fundacion Marcelino Botin. Jose Luis Saez (Ministerio de Medio Ambiente, Rural y Marino, MARM) made valuable comments on the first draft. PA is currently holding a Juan de la Cierva research contract awarded by the Ministerio de Ciencia e Innovacion – Fondo Social Europeo.

Ticks are vectors of diseases that affect humans and animals worldwide. Tick vaccines have been proposed as a cost-effective and environmentally sound alternative for tick control. Recently, the Rhipicephalus microplus Subolesin (SUB)-. Anaplasma marginale MSP1a chimeric antigen was produced in Escherichia coli as membrane-bound and exposed protein and used to protect vaccinated cattle against tick infestations. In this research, lipidomics and proteomics characterization of the E. coli membrane-bound SUB-MSP1a antigen showed the presence of components with potential adjuvant effect. Furthermore, vaccination with membrane-free SUB-MSP1a and bacterial membranes containing SUB-MSP1a showed that bacterial membranes enhance the immunogenicity of the SUB-MSP1a antigen in animal models. R. microplus female ticks were capillary-fed with sera from pigs orally immunized with membrane-free SUB, membrane bound SUB-MSP1a and saline control. Ticks ingested antibodies added to the blood meal and the effect of these antibodies on reduction of tick weight was shown for membrane bound SUB-MSP1a but not SUB when compared to control. Using the simple and cost-effective process developed for the purification of membrane-bound SUB-MSP1a, endotoxin levels were within limits accepted for recombinant vaccines. These results provide further support for the development of tick vaccines using E. coli membranes exposing chimeric antigens such as SUB-MSP1a., This research was supported by EU FP7 Grant WildTBVac and Plan Nacional I+D+i AGL2011-30041 from Ministerio de Economía y Competitividad (MINECO), Spain and FEDER. J.A. Moreno-Cid was funded by the Consejería de Educación y Ciencia, JCCM, Spain (project PEII09-0118-8907), as a recipient of a JCCM fellowship. Mariana Boadella is a recipient of a Marie Curie Intra-European Fellowship, grant number 627320.

The control of diseases shared with wildlife requires the development of strategies that will reduce pathogen transmission between wildlife and both domestic animals and human beings. This review describes and criticizes the options currently applied and attempts to forecast wildlife disease control in the coming decades. Establishing a proper surveillance and monitoring scheme (disease and population wise) is the absolute priority before even making the decision as to whether or not to intervene. Disease control can be achieved by different means, including: (1) preventive actions, (2) arthropod vector control, (3) host population control through random or selective culling, habitat management or reproductive control, and (4) vaccination. The alternative options of zoning or no-action should also be considered, particularly in view of a cost/benefit assessment. Ideally, tools from several fields should be combined in an integrated control strategy. The success of disease control in wildlife depends on many factors, including disease ecology, natural history, and the characteristics of the pathogen, the availability of suitable diagnostic tools, the characteristics of the domestic and wildlife host(s) and vectors, the geographical spread of the problem, the scale of the control effort and stakeholders’ attitudes., This work was supported by Plan Nacional I + D + i research grant AGL2011-30041 and FAU2008-00004 grants from MINECO and the EU FP7 grants APHAEA (EMIDA ERA-NET) and WildTBvac (project number 613779). The PhD students were supported by predoctoral grants from JCCM and MINECO.

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., This study is a contribution to the European project APHAEA (EMIDA ERA-NET).

The control of multihost pathogens, such as Coxiella burnetii, should rely on accurate information about the roles played by the main hosts. We aimed to determine the involvement of the red deer (Cervus elaphus) in the ecology of C. burnetii.We predicted that red deer populations from broad geographic areas within a European context would be exposed to C. burnetii, and therefore, we hypothesized that a series of factors would modulate the exposure of red deer to C. burnetii. To test this hypothesis, we designed a retrospective survey of 47 Iberian red deer populations from which 1,751 serum samples and 489 spleen samples were collected. Sera were analyzed by enzyme-linked immunosorbent assays (ELISA) in order to estimate exposure to C. burnetii, and spleen samples were analyzed by PCR in order to estimate the prevalence of systemic infections. Thereafter, we gathered 23 variables- within environmental, host, and management factors-potentially modulating the risk of exposure of deer to C. burnetii, and we performed multivariate statistical analyses to identify the main risk factors. Twenty-three populations were seropositive (48.9%), and C. burnetii DNA in the spleen was detected in 50% of the populations analyzed. The statistical analyses reflect the complexity of C. burnetii ecology and suggest that although red deer may maintain the circulation of C. burnetii without third species, the most frequent scenario probably includes other wild and domestic host species. These findings, taken together with previous evidence of C. burnetii shedding by naturally infected red deer, point at this wild ungulate as a true reservoir for C. burnetii and an important node in the life cycle of C. burnetii, at least in the Iberian Peninsula., This work was funded by EU FP7 grant ANTIGONE (278976) and CDTI (Centro para el Desarrollo Tecnológico Industrial, Spanish Ministry for the Economy and Competitiveness [MINECO]). J. A. Barasona holds an FPU predoctoral scholarship from MECD. J. P. V. Santos and J. Queirós were supported by Ph.D. grants (SFRH/BD/65880/2009 and SFRH/BD/73732/2010, respectively) from the Portuguese Science and Technology Foundation (FCT). F. Ruiz-Fons is supported by a “Ramón y Cajal” contract from MINECO.

We would like to thank the EU 7FP grant APHAEA (EMIDA ERA-NET) and the Plan Nacional l+D+i AGL2011-30041 from the Ministerio de Economia y Competitividad (MINECO), Spain, and FEDER.

Mycobacterium bovis causes animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivated M. bovis and challenge with a virulent M. bovis field strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests for in vivo TB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivated M. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. Massive M. bovis growth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased after M. bovis challenge. This information is relevant for pig production in regions that are endemic for M. bovis and for TB vaccine research., This study is a contribution to EU FP7 grant 613779 WildTBVac and to MINECO Plan Nacional grant AGL2011-30041 and FEDER.

In the Mediterranean part of the Spanish mainland, animal TB is still a major concern. This chapter summarizes the current knowledge on TB in cattle, other domestic M. tuberculosis complex (MTBC) hosts, and wildlife in Spain. It briefly addresses aspects of strain diversity and human infection, and provides an outlook on current and future research. In some Mediterranean habitats, a multihost system exists in which transmission among three wild ungulate species, cattle and other livestock, and to a lesser extent, other wildlife contributes to infection maintenance. Data from GPS and data logger collared wild and domestic M. bovis hosts, and camera trapping surveys at water holes and farms, will become available soon. Other ongoing studies are addressing aspects of M. bovis excretion and means of transmission. Two vaccines, the traditional live bacillus Calmette-Guérin (BCG) and a newly developed inactivated vaccine, are currently being tested in the field.

[Background]: Field vaccination trials with Mycobacterium bovis BCG, an attenuated mutant of M. bovis, are ongoing in Spain, where the Eurasian wild boar (Sus scrofa) is regarded as the main driver of animal tuberculosis (TB). The oral baiting strategy consists in deploying vaccine baits twice each summer, in order to gain access to a high proportion of wild boar piglets. The aim of this study was to assess the response of wild boar to re-vaccination with BCG and to subsequent challenge with an M. bovis field strain. [Results]: BCG re-vaccinated wild boar showed reductions of 75.8% in lesion score and 66.9% in culture score, as compared to unvaccinated controls. Only one of nine vaccinated wild boar had a culture-confirmed lung infection, as compared to seven of eight controls. Serum antibody levels were highly variable and did not differ significantly between BCG re-vaccinated wild boar and controls. Gamma IFN levels differed significantly between BCG re-vaccinated wild boar and controls. The mRNA levels for IL-1b, C3 and MUT were significantly higher in vaccinated wild boar when compared to controls after vaccination and decreased after mycobacterial challenge. [Conclusions]: Oral re-vaccination of wild boar with BCG yields a strong protective response against challenge with a field strain. Moreover, re-vaccination of wild boar with BCG is not counterproductive. These findings are relevant given that re-vaccination is likely to happen under real (field) conditions., Carlos Martín, Universidad de Zaragoza, kindly provided the vaccine BCG strain. Many colleagues at SaBio IREC, NEIKER and VISAVET granted valuable technical help. This is a contribution to EU FP7 grant 613779 WildTBVac and to MINECO Plan Nacional grant AGL2011-30041 and FEDER.

Individuals in natural populations are exposed to a diversity of pathogens which results in coinfections. The aim of this study was to investigate the relation between natural infection with tuberculosis (TB) due to infection by bacteria of the Mycobacterium tuberculosis complex and porcine circovirus type 2 (PCV2) in free-ranging Eurasian wild boar (Sus scrofa). Apparent prevalence for TB lesions and PCV2 infection was extremely high in all age classes, including piglets (51% for TB; 85.7% for PCV2). Modeling results revealed that the relative risk of young (less than 2 years old) wild boar to test positive to PCV2 PCR was negatively associated with TB lesion presence. Also, an interaction between TB, PCV2, and body condition was evidenced: in wild boar with TB lesions probability of being PCV2 PCR positive increased with body condition, whereas this relation was negative for wild boar without TB lesions. This study provides insight into the coinfections occurring in free-ranging host populations that are naturally exposed to several pathogens at an early age. Using TB and PCV2 as a case study, we showed that coinfection is a frequent event among natural populations that takes place early in life with complex effects on the infections and the hosts., This is a contribution to Plan Nacional I + D + i AGL2011-30041 from MINECO and ANTIGONE (Project no. 278976). Ph.D students (BBB, IDD, and JAB) were supported by predoctoral grants from JCCM and MINECO.

This is an open-access article.-- et al., This research was funded by EU FP7 grant ANTIGONE (#278976).

This is an Open Access article distributed under the terms of the Creative Commons Attribution License.-- et al., [Background]: Infections with Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex (MTC) are shared between livestock, wildlife and sporadically human beings. Wildlife reservoirs exist worldwide and can interfere with bovine tuberculosis (TB) eradication efforts. The Eurasian wild boar (Sus scrofa) is a MTC maintenance host in Mediterranean Iberia (Spain and Portugal). However, few systematic studies in wild boar have been carried out in Atlantic regions. We describe the prevalence, distribution, pathology and epidemiology of MTC and other mycobacteria from wild boar in Atlantic Spain. A total of 2,067 wild boar were sampled between 2008 and 2012. [Results]: The results provide insight into the current status of wild boar as MTC and Mycobacterium avium complex (MAC) hosts in temperate regions of continental Europe. The main findings were a low TB prevalence (2.6%), a low proportion of MTC infected wild boar displaying generalized TB lesions (16.7%), and a higher proportion of MAC infections (4.5%). Molecular typing revealed epidemiological links between wild boar and domestic – cattle, sheep and goat – and other wildlife – Eurasian badger (Meles meles) and red fox (Vulpes vulpes) – hosts. [Conclusions]: This study shows that the likelihood of MTC excretion by wild boar in Atlantic habitats is much lower than in Mediterranean areas. However, wild boar provide a good indicator of MTC circulation and, given the current re-emergence of animal TB, similar large-scale surveys would be advisable in other Atlantic regions of continental Europe., Nelson Marreros holds a ‘fellowship for prospective researcher’ from the Swiss National Science Foundation. This project was supported by INIA RTA2011-00010-00-00 (FEDER-cofinanciated) and EU FP7 grant APHAEA (EMIDA ERA-NET).

Tuberculosis (TB), a chronic disease caused by infection with the Mycobacterium tuberculosis complex, is endemic in wild boar (Sus scrofa) and red deer (Cervus elaphus) in south-central Spain. Understanding the temporal dynamics of this chronic infection requires long time series data collection over large areas. The aim of this paper was to identify the determinants of TB prevalence and severity in both species in Ciudad Real province, Spain, from 2000 to 2012. Study variables included management, population dynamics, and a range of geographical and climatological factors. The prevalence of TB in wild boar increased from 50% to 63% since the study commenced. This may be due to an increased hunting bag (a proxy for population abundance), which was correlated with TB infection rates. Low rainfall (a stochastic factor) was associated with higher individual risk of TB presence and progression, resulting in an increased proportion of severe cases of wild boar TB in dry years. This was probably a result of increased food restriction leading to a higher susceptibility to TB. In contrast, red deer TB showed an apparent stable trend, which may be a consequence of the species' higher and stable population size. Hunting management, characterized by fencing, was associated with a higher risk of TB in both wild boar and red deer, suggesting that intensive hunting management may have contributed to exacerbated TB figures. This difference was more marked in red deer than in wild boar, probably because fencing imposes less restriction on movement, population mixing and TB spread to wild boar than to deer. Our findings on TB dynamics are fundamental for assessing the impact of future disease-control actions (e.g. field vaccination). Moreover, such control plans must operate in the long term and cover large areas., This study was funded by the MINECO research project AGL2010-20730-C02-01 and JCCM PPEII 10-0262-7673.This is also a contribution to EU FP7 EMIDA (ERA-NET) Grant APHAEA on wildlife disease surveillance in Europeand to FP7 Grants ANTIGONE (project number 278976) and WildTBVac. Jose Angel Barasona enjoyed a scholarshipfrom JCCM and a contract through MINECO Grant AGL2011-30041. We thank the Agriculture Department of JCCM and its Delegation in Ciudad Real for providing data on hunting statistics and sanitary campaigns. Pelayo Acevedo was supported by a Beatriu de Pinòs fellowship funded y ‘Comissionat per a Universitats i Recerca del Departament d’Innovaciò, Universitats i Empresa’ of the Generalitat de Catalunya, and the COFUND Programme–MarieCurie Actions under 7th Framework Programme of the European Community. He is currently funded from the SFRH/BPD/90320/2012 post-doctoral grant provided bythe Portuguese Fundaçao para a Ciencia e a Tecnologia (FCT) and the European Social Fund.

et al., Besnoitia besnoiti has been reported to affect cattle, wildebeest, kudu and impala, and B. tarandi other wild ruminants (caribou, reindeer, mule deer and musk ox), causing similar characteristic clinical signs and lesions. However, both Besnoitia species have been reported in different geographical areas and the link between the sylvatic and domestic life cycles of Besnoita spp. in wild ruminants and cattle remains unknown. The aim of this study was to evaluate the presence of specific antibodies against Besnoitia spp. in wild ruminants in Spain. A wide panel of sera from red deer (Cervus elaphus) (n = 734), roe deer (Capreolus capreolus) (n = 124), chamois (Rupicapra pyrenaica) (n = 170) and mouflon (Ovis musimon) (n = 20) collected from different locations of Spain was analyzed. Beef cattle were present in all sampled areas and, interestingly, bovine besnoitiosis has been widely reported in some of them (e.g., Pyrenees and Central Spain). Sera samples were first examined with an Enzyme-Linked Immunosorbent Assay (ELISA). For red deer and roe deer, the ELISA was standardized with positive and negative control sera from several Cervidae species (100% Se and 98% Sp). Chamois and mouflon sera samples were tested with a previously reported ELISA validated for bovine sera (97% Se and 95% Sp) using protein G as a conjugate. Positive results by ELISA were confirmed a posteriori with a tachyzoite-based Western blot. Sixty-one sera samples from red deer and 17 sera samples from roe-deer were seropositive or doubtful by ELISA. All samples from mouflon were seronegative and 15 sera samples from chamois were considered doubtful. B. besnoiti exposure was only confirmed clearly by Western blot in one red deer and one roe deer from the Spanish Pyrenees where the disease is traditionally endemic. This is the first serological report of Besnoitia spp. infection carried out in European wild ruminants and the results show that specific antibodies are present at least in red deer and roe-deer. Thus, wild ruminants from endemic regions of bovine besnoitiosis should be further studied because they may be putative reservoirs of the parasite., This work was supported by a research project from the Spanish Ministry of Economy and Competitiveness (AGL 2010-20561). Daniel Gutiérrez Expósito has been financially supported by the Ministry of Economy and Competitiveness (grant no. BES-2011-043753).

Body condition is a useful measure of the nutritional status and performance in ungulates. The most widely used indicators of body condition are based on fat reserves, mainly those surrounding the kidneys. To estimate the body condition in large herbivores, researchers often collect and pool the data from both kidneys and their perirenal fat in order to compensate for potential differences between the body condition indicators obtained from the left and right kidneys. Since these differences do not seem to be properly established in wildlife, we checked for the potential differences between the body condition indicators, derived from both kidneys, in wild ungulates. In the present study, we used red deer Cervus elaphus, both under experimental and field conditions, as a model to analyse the patterns of response of the body condition indicators, calculated from left and right kidneys, to supplementary feeding, sex, age class and season. By considering two analytical approaches-kidney fat index and fat weight (including kidney weight as a covariate)-we consistently obtained similar statistical results and the same ecological interpretations by using left, right and average indicators for all the factors tested. In the experimental study, supplementary feeding had a significant effect on all the body condition indicators of red deer hinds. Under field circumstances, the condition indicators were statistically higher in females than in males and they increased with age. Sex related differences were significant in the winter. Taking into account our results, we hypothesize that information from only one kidney will be enough to assess the body condition in red deer and probably in other wild ungulates. However, a standardized sample (i.e. always using the same kidney and associated fat) should be used, while a better understanding regarding possible asymmetries in the amount of the energy reserves on each side of the animals' bodies has not been reached. Our study has clear implications concerning wildlife monitoring since it represents a considerable reduction of time and effort, both in the field (collecting the samples) and in the laboratory (processing and weighing the samples) when evaluating the body condition of individuals and populations., This study was supported by EU FP7 grant APHAEA (EMIDA ERA-NET) and CDTI. João P.V. Santos was supported by a PhD Grant (SFRH/BD/65880/2009) from Fundação para a Ciência e a Tecnologia (FCT) co-financed by the European Social Fund POPH-QREN programme. Pelayo Acevedo was supported by a Beatriu de Pinós fellowship funded by Comissionat per a Universitats i Recerca del Departament d’Innovació, Universitats i Empresa, of the Generalitat de Catalunya, and the COFUND Programme – Marie Curie Actions under the 7th Framework Programme of the European Community.

[ES]: Las especies cinegéticas comparten infecciones con el hombre, los animales domésticos y el resto de la fauna silvestre que pueden afectar seriamente a la salud pública, la sanidad animal, la conservación y la producción cinegética; de ahí su relevancia. Pese a ello, la sanidad de las especies cinegéticas apenas es tenida en cuenta en los planes de gestión de estas especies. En este texto enumeramos las principales debilidades de la gestión cinegética desde el punto de vista sanitario: (1) la no aplicación de la legislación sobre control sanitario en traslados y repoblaciones; (2) la indefinición del estatus de las granjas cinegéticas; (3) la ausencia de un registro de “cercones” y la ilegalidad de los mismos; (4) la falta de clasificación correcta de los terrenos cinegéticos; (5) el seguimiento inadecuado del tamaño de las poblaciones y sus efectos sobre el medio; (6) la falta de regulación del aporte artificial de alimento; (7) la falta de consideración de los riesgos sanitarios derivados de la sobreabundancia; (8) los conflictos por la gestión de residuos de caza; (9) la deficiente inspección sanitaria de los animales cazados; (10) la no regulación de los tratamientos farmacológicos en especies cinegéticas y (11) la falta de intercambio de información dentro de la Administración Pública y la necesidad de valoración y aprobación del plan técnico de caza por la autoridad de sanidad animal. Para cada una de ellas, proponemos cambios en la regulación de la actividad cinegética, basándonos en ejemplos españoles y en la literatura científica internacional., [EN]: Game species share diseases with humans, livestock and wildlife, hence their relevance. However, game species health is not considered in current Spanish hunting management regulation. This paper lists the main weaknesses of game management from the sanitary point of view: (1) insufficient enforcement of current animal health and movement regulations, (2) lack of a clear definition and classification of game farms, (3) uncertain status of interior wild boar fences or “cercones”, (4) lack of a proper classification of the hunting estates, (5) inadequate wildlife population monitoring, (6) lack of regulation on artificial feeding, (7) health risks associated to overabundance; (8) big game waste conflicts (9) poor veterinary inspection of hunted animals (10), lack of regulation of pharmaceutical treatments on game species, and (11) poor information flow within the administration departments. For each point, we propose changes to the hunting regulation, based on Spanish and international scientific literature., Este trabajo es una contribución a la Encomienda de gestión sobre vigilancia sanitaria de fauna silvestre de MAGRAMA a IREC-CSIC, y al proyecto europeo APHAEA (EMIDA ERA-NET). Marina Álvarez (MAGRAMA) aportó información valiosa sobre las granjas cinegéticas registradas en España. PA disfruta actualmente de un contrato postdoctoral (SFRH/BPD/90320/2012) de la Fundação para a Ciência e a Tecnologia.

This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al., 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, This study was funded by our own institutional resources (Anses).

This is an Open Access article distributed under the terms of the Creative Commons Attribution License.-- et al., [Background]: Anaplasma phagocytophilum infects a wide variety of hosts and causes granulocytic anaplasmosis in humans, horses and dogs and tick-borne fever in ruminants. Infection with A. phagocytophilum results in the modification of host gene expression and immune response. The objective of this research was to characterize gene expression in pigs (Sus scrofa) naturally and experimentally infected with A. phagocytophilum trying to identify mechanisms that help to explain low infection prevalence in this species. [Results]: For gene expression analysis in naturally infected pigs, microarray hybridization was used. The expression of differentially expressed immune response genes was analyzed by real-time RT-PCR in naturally and experimentally infected pigs. Results suggested that A. phagocytophilum infection affected cytoskeleton rearrangement and increased both innate and adaptive immune responses by up regulation of interleukin 1 receptor accessory protein-like 1 (IL1RAPL1), T-cell receptor alpha chain (TCR-alpha), thrombospondin 4 (TSP-4) and Gap junction protein alpha 1 (GJA1) genes. Higher serum levels of IL-1 beta, IL-8 and TNF-alpha in infected pigs when compared to controls supported data obtained at the mRNA level. [Conclusions]: These results suggested that pigs are susceptible to A. phagocytophilum but control infection, particularly through activation of innate immune responses, phagocytosis and autophagy. This fact may account for the low infection prevalence detected in pigs in some regions and thus their low or no impact as a reservoir host for this pathogen. These results advanced our understanding of the molecular mechanisms at the host-pathogen interface and suggested a role for newly reported genes in the protection of pigs against A. phagocytophilum., This research was supported by EU FP7, ANTIGONE project number 278976 and the CSIC intramural project 200830I249 to JF. N. Ayllón was funded by MINECO, Spain.

Fallow deer (Dama dama) are widely distributed as natural or naturalised populations, as well as in game parks and deer farms. We used 157 fallow deer sampled in populations considered to be Mycobacterium tuberculosis complex (MTC) free and 73 Mycobacterium bovis-infected fallow deer confirmed postmortem by culture to evaluate the diagnostic performance of two tests for the detection of anti-mycobacterial antibodies: the dual path platform (DPP) VetTB assay and the bovine purified protein derivative (bPPD) ELISA. We also compared their sensitivity with that of the skin test, analyzed the effect of haemolysis degree on the antibody detection and described the relationship between the test readings and presence/absence of gross tuberculosis (TB) compatible lesions. Sensitivity of bPPD ELISA was 51% at a specificity of 96%. Depending on the cut-off value selected, the sensitivity of DPP VetTB ranged from 62 to 71%, while its specificity was 88-95%. In the subgroup of M. bovis-infected deer for which the skin test data were available (33 of 73); this method detected 76% of culture-positive animals, although the specificity of the intradermal test was not determined in this study. When the DPP VetTB and skin test data were combined, the resulting sensitivity obtained in this sub-group of M. bovis-infected deer increased to 97%. Gross pathology identified TB compatible lesions (TBL) in 89% culture-confirmed fallow deer. The infected animals with visible lesions had significantly higher readings in the DPP VetTB, but not in the bPPD ELISA. Only high levels of haemolysis decreased antibody test sensitivity and this effect was more evident for the bPPD ELISA. The results allowed inferring a number of management recommendations for rapid detection of MTC infection in live fallow deer and in surveys on hunter-harvested cervids., This is a contribution to MCINN Plan Nacional Research grant AGL2008-03875 and FEDER, and to EU grant TB-STEP 212414. Studies on diseases shared between domestic animals and wildlife are also supported by grants and contracts from INIA and Ministerio de Medio Ambiente y Medio Rural y Marino (SDGSPP), and Grupo Santander – Fundación Marcelino Botín.