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Review. et al. ; Many infectious diseases originating from, or carried by, wildlife affect wildlife conservation and biodiversity, livestock health, or human health. We provide an update on changes in the epidemiology of 25 selected infectious, wildlife-related diseases in Europe (from 2010–16) that had an impact, or may have a future impact, on the health of wildlife, livestock, and humans. These pathogens were selected based on their: 1) identification in recent Europe-wide projects as important surveillance targets, 2) inclusion in European Union legislation as pathogens requiring obligatory surveillance, 3) presence in recent literature on wildlife-related diseases in Europe since 2010, 4) inclusion in key pathogen lists released by the Office International des Epizooties, 5) identification in conference presentations and informal discussions on a group email list by a European network of wildlife disease scientists from the European Wildlife Disease Association, or 6) identification as pathogens with changes in their epidemiology during 2010–16. The wildlife pathogens or diseases included in this review are: avian influenza virus, seal influenza virus, lagoviruses, rabies virus, bat lyssaviruses, filoviruses, canine distemper virus, morbilliviruses in aquatic mammals, bluetongue virus, West Nile virus, hantaviruses, Schmallenberg virus, Crimean-Congo hemorrhagic fever virus, African swine fever virus, amphibian ranavirus, hepatitis E virus, bovine tuberculosis (Mycobacterium bovis), tularemia (Francisella tularensis), brucellosis (Brucella spp.), salmonellosis (Salmonella spp.), Coxiella burnetii, chytridiomycosis, Echinococcus multilocularis, Leishmania infantum, and chronic wasting disease. Further work is needed to identify all of the key drivers of disease change and emergence, as they appear to be influencing the incidence and spread of these pathogens in Europe. We present a summary of these recent changes during 2010–16 to discuss possible commonalities and drivers of disease change and to identify directions for future work on wildlife-related diseases in Europe. Many of the pathogens are entering Europe from other continents while others are expanding their ranges inside and beyond Europe. Surveillance for these wildlife-related diseases at a continental scale is therefore important for planet-wide assessment, awareness of, and preparedness for the risks they may pose to wildlife, domestic animal, and human health. ; F.R.F. was funded by the Spanish Ministry for the Economy, Industry and Competitiveness (MINECO) through the 'Ramon y Cajal program. ; Peer reviewed

We studied the role of European rabbits (Oryctolagus cuniculus) as a reservoir for Coxiella burnetii in the Iberian region. High individual and population seroprevalences observed in wild and farmed rabbits, evidence of systemic infections, and vaginal shedding support the reservoir role of the European rabbit for C. burnetii. ; This study was supported by European Union FP7 grant ANTIGONE (278976), European Union FP7 EMIDA ERA-NET grant APHAEA on wildlife disease surveillance in Europe, and Centro para el Desarrollo Tecnológico Industrial, Spanish Ministry for Economy and Competitiveness. F.-R-F. was supported by Juan de la Cierva and Ramón y Cajal contracts from the Spanish Ministry for Economy and Competitiveness. ; Peer Reviewed

We studied the potential of red deer as bluetongue maintenance hosts and sentinels. Deer maintained detectable bluetongue virus (BTV) serotype 4 RNA for 1 year after the virus was cleared from livestock. However, the virus was not transmitted to yearlings. BTV serotype 1 RNA was detected in red deer immediately after its first detection in cattle. ; This study was funded by the Spanish Ministry of Natural, Rural and Marine Environment (RASVE 274/2007, and an agreement between Organismo Autónomo de Parques Nacionales (OAPN), Dirección General de Recursos Agricolas y Ganaderos (DGRAG), and Consejo Superior de Investigaciones Cientificas (CSIC). F.R.-F. is supported by a postdoctoral contract of the Instituto de Salud Carlos III of the Spanish government. ; Peer Reviewed

More than one hundred years ago African swine fever (ASF) was first diagnosed in Kenya. Since then, diverse approaches have been applied to the study of the causative virus, the sole member of the family Asfarviridae, aimed at characterising its properties, genome organisation and replication, its antigenic and biological properties as well as to develop treatment and a vaccine. The disease evolved and has persisted in Africa in a sylvatic cycle involving wild suids and soft ticks for a long time, but was introduced, usually through contaminated waste food, into other regions on multiple occasions since 1957. The most recent introduction, into Georgia in 2007, resulted in the spread of the disease to the European Union in 2014 and to the establishment of an international and multidisciplinary network of scientists funded by the European Cooperation in Science and Technology (COST) two years later. The network included a broad variety of scientific fields, animal health and food safety authorities, hunting associations, wildlife managers and food and livestock industries with the goal of increasing preparedness and attempting to stop ASF spread. This book represents the summary of the collective and integrated work of almost 300 dedicated participants in tackling the complex challenge posed by ASF. Here we summarise the state-of-the-art knowledge on this lethal disease, with a focus on the European situation, and identify areas that still need to be explored. ; This publication is based on work from 'Understanding and combating African swine fever in Europe (ASF-STOP COST action 15116)' supported by COST (European Cooperation in Science and Technology). LI received funding from the Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Action (Grant Agreement no. 656697). ; Peer reviewed

Foci of tick-borne pathogens occur at fine spatial scales, and depend upon a complex arrangement of factors involving climate, host abundance and landscape composition. It has been proposed that the presence of hosts that support tick feeding but not pathogen multiplication may dilute the transmission of the pathogen. However, models need to consider the spatial component to adequately explain how hosts, ticks and pathogens are distributed into the landscape. ; In this study, a novel, lattice-derived, behavior-based, spatially-explicit model was developed to test how changes in the assumed perception of different landscape elements affect the outcome of the connectivity between patches and therefore the dilution effect. The objective of this study was to explain changes in the exposure rate (ER) of red deer to Anaplasma spp. under different configurations of suitable habitat and landscape fragmentation in the presence of variable densities of the potentially diluting host, wild boar. The model showed that the increase in habitat fragmentation had a deep impact on Habitat Sharing Ratio (HSR), a parameter describing the amount of habitat shared by red deer and wild boar, weighted by the probability of the animals to remain together in the same patch (according to movement rules), the density of ticks and the density of animals at a given vegetation patch, and decreased the dilution effect of wild boar on deer Anaplasma ER. ; The model was validated with data collected on deer, wild boar and tick densities, climate, landscape composition, host vegetation preferences and deer seropositivity to Anaplasma spp. (as a measure of ER) in 10 study sites in Spain. However, although conditions were appropriate for a dilution effect, empirical results did not show a decrease in deer ER in sites with high wild boar densities. The model showed that the HSR was the most effective parameter to explain the absence of the dilution effect. These results suggest that host habitat usage may weaken the predicted dilution effect for tick-borne pathogens and emphasize the importance of the perceptual capabilities of different hosts in different landscapes and habitat fragmentation conditions for predictions of dilution effects. ; This research was supported by grants "Epidemiología de zoonosis transmitidas por garrapatas en Castilla La Mancha" (06036-00 ICS-JCCM), Epidemiología de las enfermedades compartidas entre ungulados silvestres y ganado en España: aproximación multidisciplinar (MEC AGL2005-07401) and FEDER and was facilitated through the Integrated Consortium on Ticks and Tick-borne Diseases (ICTTD-3), financed by the International Cooperation Program of the European Union, coordination action project No. 510561. PA has a postdoc grant from the Junta de Comunidades de Castilla-La Mancha, Spain. Contributions intended to support the filed works and data collection. Sponsors are public and have no role in any activity. ; Peer reviewed

[Background]: Q fever, a worldwide zoonotic disease caused by Coxiella burnetii, is endemic in northern Spain where it has been reported as responsible for large series of human pneumonia cases and domestic ruminants' reproductive disorders. To investigate pathogen exposure among domestic ruminants in semi-extensive grazing systems in northern Spain, a serosurvey was carried out in 1,379 sheep (42 flocks), 626 beef cattle (46 herds) and 115 goats (11 herds). Serum antibodies were analysed by ELISA and positive samples were retested by Complement Fixation test (CFT) to detect recent infections. [Results]: ELISA anti-C. burnetii antibody prevalence was slightly higher in sheep (11.8 ± 2.0%) than in goats (8.7 ± 5.9%) and beef cattle (6.7 ± 2.0%). Herd prevalence was 74% for ovine, 45% for goat and 43% for bovine. Twenty-one percent of sheep flocks, 27% of goat and 14% of cattle herds had a C. burnetii seroprevalence ¿ 20%. Only 15 out of 214 ELISA-positive animals reacted positive by CFT. Age-associated seroprevalence differed between ruminant species with a general increasing pattern with age. No evidence of correlation between abortion history and seroprevalence rates was observed despite the known abortifacient nature of C. burnetii in domestic ruminants. [Conclusions]: Results reported herein showed that sheep had the highest contact rate with C. burnetii in the region but also that cattle and goats should not be neglected as part of the domestic cycle of C. burnetii. This work reports basic epidemiologic patterns of C. burnetii in semi-extensive grazed domestic ruminants which, together with the relevant role of C. burnetii as a zoonotic and abortifacient agent, makes these results to concern both Public and Animal Health Authorities. ; This work was supported by funding from INIA (FAU2006-00002-C04-01), FEDER, and the Department of Agriculture, Fisheries and Food of the Basque Government. F. Ruiz-Fons is supported by a research contract from the 'Instituto de Salud Carlos III' of the Spanish Ministry for Health. ; Peer Reviewed

This is an Open Access article distributed under the terms of the Creative Commons Attribution License.-- et al. ; The diversity of Babesia species infecting cervids in parts of central and southern Spain was analyzed by collecting blood from farmed red deer (Cervus elaphus). Babesia sp. was isolated in vitro from two red deer herds in Cádiz and Ciudad Real. The number of Babesia sp. carriers differed between the two herds: 36/77 in Cádiz and 1/35 in Ciudad Real. Hyalomma lusitanicum was the most prevalent tick species identified on the Cádiz farm vegetation and on sampled animals, and is therefore a candidate vector. The molecular characteristics of 21 isolates were determined by complete (8 isolates) or partial (13 isolates) 18S rRNA gene sequencing. The sequences were highly similar (over 99.4% identity) and 6 sequence types were identified at the level of one herd only, demonstrating a rather high genetic diversity. They formed a monophyletic clade, and members of the three main sequence types shared a similar morphology and the same erythrocyte susceptibility pattern. This clade also included Babesia sp. Xinjiang isolated from sheep in China and Babesia sp. identified in giraffe in South Africa, with identities higher than 98.3% and statistically relevant phylogenetic support. None of the biological properties analyzed for both Babesia from red deer and Babesia sp. Xinjiang allowed their differentiation (ability to develop in vitro in erythrocytes from cattle and sheep, as well as in erythrocytes from different cervids, unsuccessful infection of calves). We propose the Babesia isolated from red deer as a new species named B. pecorum. Whether Babesia sp. Xinjiang and the Babesia characterized in South Africa belong to the same species is debated. ; This research was supported by funds from the Institut National de la Recherche Agronomique, the European Union Cooperation Project PIROVAC (KBBE-3-245145) and from the project "Zoonosis transmitidas por vectores hematófagos en Castilla-La Mancha: Mapa de riesgos" (JCCM POII09-0141-8176) and CDTI/Medianilla. F. Ruiz-Fons and I.G. Fernández de Mera are supported by a Juan de la Cierva contract from the Spanish Ministry for Economy and Competitiveness. ; Peer Reviewed

Bulk tank milk (BTM) samples were collected from 81 sheep flocks in the Basque Country, Spain, in 2015 and were analysed for antibodies against Coxiella burnetii by ELISA and for C. burnetii DNA by real-time PCR. Thirty-two percent of the flocks had BTM antibodies against C. burnetii. Presence of C. burnetii DNA in BTM was detected in 23% of the flocks, suggesting recent C. burnetii infections. Retrospective data of BTM samples obtained from 154 sheep flocks investigated in 2005 in the same geographic area were compiled to assess temporal changes in C. burnetii infection. The overall percentage of infected sheep flocks did not significantly change after the 10-year period. Among the 46 flocks sampled in both periods, 11 flocks that were negative in 2005 were positive in 2015, 18 maintained their initial status (positive or negative), and 17 positive flocks were negative in 2015. These findings indicate that C. burnetii infection is a dynamic process in dairy sheep in northern Spain. Single nucleotide polymorphism (SNP) genotyping of positive samples identified three genotypes, SNP1 being the most prevalent in 2015 and SNP8 in 2005; SNP4 was only detected once in 2005. These results suggest possible changes in the pattern of genotype infection over time. ; This work was funded by INIA (RTA2013-00051-C02-00), FEDER funds and the Basque Government. RAA is beneficiary of a Ph.D. contract funded by INIA (FPI-2015-014). FRF is funded by the Spanish Ministry for the Economy, Industry and Competitiveness (MINECO) through a contract under the Ramón y Cajal program. ; Peer Reviewed

Bulk tank milk (BTM) samples were collected from 81 sheep flocks in the Basque Country, Spain, in 2015 and were analysed for antibodies against Coxiella burnetii by ELISA and for C. burnetii DNA by real-time PCR. Thirty-two percent of the flocks had BTM antibodies against C. burnetii. Presence of C. burnetii DNA in BTM was detected in 23% of the flocks, suggesting recent C. burnetii infections. Retrospective data of BTM samples obtained from 154 sheep flocks investigated in 2005 in the same geographic area were compiled to assess temporal changes in C. burnetii infection. The overall percentage of infected sheep flocks did not significantly change after the 10-year period. Among the 46 flocks sampled in both periods, 11 flocks that were negative in 2005 were positive in 2015, 18 maintained their initial status (positive or negative), and 17 positive flocks were negative in 2015. These findings indicate that C. burnetii infection is a dynamic process in dairy sheep in northern Spain. Single nucleotide polymorphism (SNP) genotyping of positive samples identified three genotypes, SNP1 being the most prevalent in 2015 and SNP8 in 2005; SNP4 was only detected once in 2005. These results suggest possible changes in the pattern of genotype infection over time. ; This work was funded by INIA (RTA2013-00051-C02-00), FEDER funds and the Basque Government. RAA is beneficiary of a Ph.D. contract funded by INIA (FPI-2015-014). FRF is funded by the Spanish Ministry for the Economy, Industry and Competitiveness (MINECO) through a contract under the Ramón y Cajal program. ; Sí

Letter to the Editor. ; F. R.-F. and I.G.F.M. are supported by a Juan de la Cierva contract from the Spanish Ministry for Economy and Competitiveness. Research supported by POII09-0141-8176 and European Union FP7 ANTIGONE (Anticipating the Global Onset of Novel Epidemics) project number 278976. ; Peer Reviewed

Animal tuberculosis (TB) remains a major problem in some countries despite the existence of control programmes focused mainly on cattle. In this species, aerogenous transmission is accepted as the most frequent infection route, affecting mainly the respiratory system. Under the hypothesis that the oral route could be playing a more relevant role in transmission, diagnosis and disease persistence than previously thought, this study was performed to assess the course of TB infection in cattle and its effects on diagnosis depending on the route of entry of Mycobacterium bovis. Two groups of five calves each were either endotracheally (EC) or orally (OC) challenged. Necropsies were carried out 12 weeks after challenge except for three OC calves slaughtered 8 weeks later. All animals reacted to the tuberculin skin test and the entire EC group was positive to the interferon-gamma release assay (IGRA) 2 weeks after challenge and thereafter. The first positive IGRA results for OC calves (3/5) were recorded 4 weeks after challenge. Group comparison revealed significant differences in lesion and positive culture location and scoring. TB-compatible gross lesions and positive cultures were more frequently found in the thorax (p < 0.001) and lung (p < 0.05) of EC animals, whereas OC animals presented lesions (p = 0.23) and positive cultures (p < 0.05) mainly located in the abdomen. These results indicate that the infection route seems to be a determining factor for both the distribution and the time needed for the development of visible lesions. Our study suggests that confirmation of TB infection in some skin reactor animals can be problematic if current post-mortem examination and diagnostics are not improved. ; This study was supported with funds from the Spanish Ministry of Economy and Competitiveness (Research Project AGL2014-56305-C3-3-R) and the Department of Economic Development and Competitiveness of the Basque Government. MS holds a fellowship from the Department of Education of the Basque Government (PRE_2017_2_0043). ; Peer Reviewed

43 pages, 3 figures, 5 tables, 1 additional file.-- et al. ; [Background]: The role of wildlife as a brucellosis reservoir for humans and domestic livestock remains to be properly established. The aim of this work was to determine the aetiology, apparent prevalence, spatial distribution and risk factors for brucellosis transmission in several Iberian wild ungulates. ; [Methods]: A multi-species indirect immunosorbent assay (iELISA) using Brucella S-LPS antigen was developed. In several regions having brucellosis in livestock, individual serum samples were taken between 1999 and 2009 from 2,579 wild bovids, 6,448 wild cervids and 4,454 Eurasian wild boar (Sus scrofa), and tested to assess brucellosis apparent prevalence. Strains isolated from wild boar were characterized to identify the presence of markers shared with the strains isolated from domestic pigs. ; [Results] Mean apparent prevalence below 0.5% was identified in chamois (Rupicapra pyrenaica), Iberian wild goat (Capra pyrenaica), and red deer (Cervus elaphus). Roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis aries) and Barbary sheep (Ammotragus lervia) tested were seronegative. Only one red deer and one Iberian wild goat resulted positive in culture, isolating B. abortus biovar 1 and B. melitensis biovar 1, respectively. Apparent prevalence in wild boar ranged from 25% to 46% in the different regions studied, with the highest figures detected in South-Central Spain. The probability of wild boar being positive in the iELISA was also affected by age, age-by-sex interaction, sampling month, and the density of outdoor domestic pigs. A total of 104 bacterial isolates were obtained from wild boar, being all identified as B. suis biovar 2. DNA polymorphisms were similar to those found in domestic pigs. ; [Conclusions]: In conclusion, brucellosis in wild boar is widespread in the Iberian Peninsula, thus representing an important threat for domestic pigs. By contrast, wild ruminants were not identified as a significant brucellosis reservoir for livestock. ; This is a contribution to the MCINN Plan Nacional research grant AGL2005-07401 on shared diseases and FEDER. The study benefited from agreements of IREC with MARM-OAPN, Castilla – La Mancha and Principado de Asturias. Additional support to the IREC is acknowledged to FISCAM (GC05-006 and PI-2007/56). CITA and UNIZAR also acknowledge support from INIA (FAU2008-00015). The Aragon Government has financed part of this work under the programme "Health status surveillance on game wildlife in Aragon". NEIKER thanks the funding of the Department for Environment, Spatial Planning, Agriculture and Fisheries of the Basque Government and the collaboration of ACCA and Regional Governments. Grant and postdoctoral contract acknowledgements: P.M Muñoz (CITA Technologist grant and Juan de la Cierva research contract), M. Boadella (PhD grant TB-STEP, FP7). P. Acevedo (Juan de la Cierva research contract, MICINN and FEDER, project CGL2006-09567/BO). F. Ruiz-Fons (I.S. Carlos III research contract, Spanish Ministry of Health). ; Peer reviewed

Tick-borne infectious diseases and allergies are a growing problem worldwide. Tick bite allergy has been associated with the direct effect of immunoglobulin E (IgE) response to tick salivary antigens, or secondary to the induction of allergy to red meat consumption through IgE antibodies against the carbohydrate α-Gal (Gal α 1-3Gal β 1-(3)4GlcNAc-R). However, despite the growing burden of this pathology, the proteins associated with anaphylaxis to tick bite have not been characterized. To address this question, a comparative proteomics approach was used to characterize tick proteins producing an IgE antibody response in a healthy individual with record of tick bites, which had not resulted in any allergic reactions, and two patients with anaphylactic reactions to Rhipicephalus bursa or Hyalomma marginatum tick bites. Both patients and the healthy individual were red meat tolerant. The results supported a patientspecific IgE antibody response to tick species responsible for the anaphylaxis to tick bite. Both patients and the healthy individual serologically recognized tick proteins with and without a-Gal modifications, with proteins differentially recognized by patients but not control sera. These proteins could be used as potential antigens for diagnostics, treatment and prevention of tick bite-induced allergies. ; Part of this research was supported by the European Union FP7 ANTIGONE project number 278976. L.M-H was supported by the University of Castilla La Mancha (UCLM, Spain) fellowship. M.V and I.G.FM were supported by the Research Plan of UCLM. F.R-F was supported by the 'Ramón y Cajal' program of the Spanish Ministry of Economy and Competitiveness. ; Peer Reviewed

In press. ; Crimean-Congo haemorrhagic fever virus (CCHFV) continues to cause new human cases in Iberia while its spatial distribution and ecological determinants remain unknown. The virus remains active in a silent tick-animal cycle to which animals contribute maintaining the tick populations and the virus itself. Wild ungulates, in particular red deer, are essential hosts for Hyalomma ticks in Iberia, which are the principal competent vector of CCHFV. Red deer could be an excellent model to understand the ecological determinants of CCHFV as well as to predict infection risks for humans because it is large, gregarious, abundant and the principal host for Hyalomma lusitanicum. We designed a cross-sectional study, analysed the presence of CCHFV antibodies in 1444 deer from 82 populations, and statistically modelled exposure risk with host and environmental predictors. The best-fitted statistical model was projected for peninsular Spain to map infection risks. Fifty out of 82 deer populations were seropositive, with individual population prevalence as high as 88%. The highest prevalence of exposure to CCHFV occurred in the southwest of the Iberian Peninsula. Climate and ungulate abundance were the most influential predictors of the risk of exposure to the virus. The highest risk regions were those where H. lusitanicum is most abundant. Eight of the nine primary human cases occurred in or bordering these regions, demonstrating that the model predicts human infection risk accurately. A recent human case of CCHF occurred in northwestern Spain, a region that the model predicted as low risk, pointing out that it needs improvement to capture all determinants of the CCHFV infection risk. In this study, we have been able to identify the main ecological determinants of CCHFV, and we have also managed to create an accurate model to assess the risk of CCHFV infection. ; This study has been funded by the Spanish Ministry for the Science and Innovation (MCI) through projects CGL2017-89866-R & E-RTA2015-0002-C02-02, and by the Regional Government of Castilla-La Mancha and the European Social Fund (ESF) through project SBPLY/19/180501/000321. RC-M, LC-M, SJ-R and JM-G would like to thank the MCI, ESF and the University of Castilla-La Mancha for their support through contracts PRE2018-083801, PEJ2018-003155-A, 2018/12504 and BES-2015-072206, respectively. BC was supported by FCT-Fundação para a Ciência e Tecnologia (2020.04872.BD). DG-B would like to thank MCI and Instituto de Salud Carlos III (CD19CIII/00011) for funding him. ; Peer reviewed