Prevalencia de patógenos zoonósicos (virus de Influenza Aviar, flavivirus del Complejo Antigénico de la Encefalitis Japonesa y Salmonella spp.) y resultados preliminares de parasitofauna hemática en paloma doméstica (Columba livia doméstica) y animales de la colección del Parque zoológico Municipal de Córdoba

Premio extraordinario de Trabajo Fin de Máster curso 2013-2014. Medicina,Sanidad y Mejora Animal.Durante el periodo 2013-2014 se realizó un estudio transversal para determinar la prevalencia de patógenos zoonósicos y hemoparásitos en 154 palomas domésticas (Columba livia var. domestica) capturadas en el Parque Zoológico Municipal de Córdoba (PZMC) y en 93 muestras de animales de la colección del Parque. Las muestras de suero se analizaron para detectar anticuerpos frente a virus de la Influenza Aviar (VIA) y flavivirus del Complejo Antigénico de la Encefalitis Japonesa (CAEJ). Así mismo, se analizaron muestras de contenidos digestivos y heces para el aislamiento de Salmonella spp. Paralelamente, se determinó la frecuencia de infección de hemoparásitos en 60 palomas. No se detectaron anticuerpos frente a VIA en ninguna de las 148 (0,0%; IC95%:0,0- 1,9) palomas analizadas mediante bELISA, mientras que se confirmó seropositividad en 5 de 28 (18,5%; IC95%:6,0-31,0) aves de la colección. Once de 142 (8,5%; IC95%:4,0-12,9) sueros de palomas presentaron anticuerpos frente a flavivirus del CAEJ mediante bELISA. Además, 4 de los 49 (8,2%; IC95%:1,9-14,5) ejemplares de la colección analizados resultaron seropositivos frente a flavivirus. El cultivo para el aislamiento de Salmonella spp. determinó ausencia de infección en las 152 muestras de contenido digestivo de paloma (0,0%; IC95%:0,0-1,9), mientras que 4 de 44 (9,1%; IC95%:1,0-17,2) muestras de heces de los animales de la colección resultaron positivas. Haemoproteus sub. Haemoproteus spp. fue detectado en 52 de las 60 (86,7%; IC95%:78,3-95,0) palomas analizadas. Los resultados obtenidos indican circulación de flavivirus del CAEJ en la población de palomas y animales del PZMC. Estas especies podrían ser usadas como centinelas para la monitorización de flavivirus en zonas urbanas. La ausencia de positividad frente a VIA y Salmonella spp., sugieren que las palomas no tienen un papel relevante en la epidemiología de estos patógenos. Los resultados determinan una elevada diseminación de Haemoproteus (H.) en las poblaciones de palomas del PZMC, lo cual podría tener implicación para la conservación de algunas especies de aves.A cross-sectional study was carried out to determine the prevalence of different pathogenic zoonotic agents and hemoparasites in 154 domestic pigeon (Columba livia var. domestica) and 93 samples of animals from the Córdoba Municipal Zoo Park (CMZP) between 2013 and 2014. Serum samples were tested to determine the presence of antibodies against Avian Influenza virus (AIV) and flaviviruses of the Japanese Encephalitis Antigenic Complex (JEAC). Moreover, samples of digestive content were analyzed for Salmonella spp. isolation. Seropositivity to AIV was not found in any of the 148 pigeons samples tested, while 5 out of 28 (18.5%; IC95%:6.0-31.0) zoo birds showed positive results using bELISA. A total of 11 out of 142 (8.5%; IC95%:4.0-12.9) pigeons analyzed presented antibodies to JEAC by bELISA. Thus, 4 out of 49 (8.2%; IC95%:1.9-14.5) zoo animals were seropositive against flaviviruses. Salmonella spp. was not isolated in any of the 152 intestine samples of pigeon (0.0%; IC95%:0.0-1.9), but 4 out of 44 samples (9.1%; IC95%:1.0-17.2) from zoo animals were positive. Finally, 52 out of 60 (86.7%; IC95%:78.3-95.0) were infected by Haemoproteus sub. Haemoproteus spp. The results obtained in the present study indicate circulation of flaviviruses of the JEAC in domestic pigeons and animals from the CMZP. The absence of seropositivity against AIV and the negative cultures to Salmonella spp. in the pigeons tested, suggest a limited role of this species in the epidemiology of both diseases. Parasitological results suggest a widespread of Haemoproteus (H.) in the pigeon population, which could has implications for the conservation of bird species

Epidemiological study of zoonotic diseases from a One Health approach

El concepto One Health (una sola salud) busca promover, mejorar y defender la salud y el bienestar de todas las especies mediante la cooperación y colaboración entre profesionales de Salud Pública, Sanidad Animal y Salud Ambiental. Dicho concepto está experimentando un resurgir importante en las últimas décadas debido al creciente interés sobre las relaciones epidemiológicas de las enfermedades compartidas entre el ser humano, los animales domésticos, la fauna silvestre y el medioambiente, y a la evidencia de que la lucha frente a estas enfermedades ha de abordarse desde una perspectiva holística. El objetivo general de la presente Tesis Doctoral es evaluar el papel de diferentes especies domésticas, peridomésticas y silvestres en la epidemiología de enfermedades zoonósicas bacterianas, víricas y parasitarias desde una perspectiva One Health. En el primer estudio (Capítulo 1.1) se estableció la seroprevalencia, factores de riesgo, distribución espacial y espoligotipos del complejo Mycobacterium tuberculosis (CMT) circulantes en cerdos criados en extensivo en Andalucía, para evaluar su papel en el manteniendo y la transmisión de la tuberculosis (TB). La seroprevalencia individual de CMT fue del 2,3% (82/3.622) y la prevalencia de explotación del 24,8% (32/129). El censo porcino y la presencia de explotaciones caprinas colindantes fueron factores de riesgo asociados con la seropositividad de CMT en las granjas porcinas. Se identificaron dos clústeres espaciales estadísticamente significativos (P<0,001) y se aislaron un total de 25 espoligotipos de CMT diferentes en cerdos criados en extensivo en la zona de estudio, compartidos con otras especies domésticas y silvestres. La baja seroprevalencia individual encontrada sugiere que los cerdos ibéricos podrían actuar como hospedadores accidentales del CMT. Sin embargo, la alta prevalencia de rebaño, la identificación de clústeres espaciales y la detección de espoligotipos previamente aislados en diferentes especies, indican la necesidad de implementar programas de control en las explotaciones de cerdo manejadas bajo sistemas de producción extensivos en España. El objetivo del Capítulo 1.2 fue evaluar la utilidad de la correcta eliminación de los residuos de caza mayor en áreas de alta prevalencia de TB como medida de control del CMT en jabalí (Sus scrofa) y ciervo (Cervus elaphus) (considerados reservorios naturales del CMT en España), durante las temporadas de caza 2008/2009 a 2016/2017. Los resultados evidenciaron que la eliminación adecuada de los subproductos de caza contribuyó a reducir la seroprevalencia del CMT en jabalí en un 25%. La correcta eliminación de los residuos de la caza mayor ha logrado reducir la seroprevalencia de tuberculosis en jabalí (Sus scrofa) hasta en un 25%, siendo por tanto una medida eficaz para control de tuberculosis en esta especie. Estos resultados son de especial relevancia en el contexto actual de enfermedades emergentes y reemergentes como la TB y la peste porcina africana en Europa. El objetivo del Capítulo 2.1 fue determinar la prevalencia de agentes zoonósicos (flavivirus, virus de la influenza aviar (VIA), Salmonella spp. y Toxoplasma gondii) en palomas y especies simpátricas del parque zoológico de Córdoba (sur de España) entre 2013 y 2014. En este estudio, se detectaron anticuerpos frente a flavivirus en 7,8% de 142 palomas y en el 8,2% de los 49 animales de zoológico analizados. Aunque no se encontró seropositividad frente a VIA ni infección por Salmonella spp. en palomas, el 17,9% (5/28) y el 6,8% (3/44) de los animales de zoológico examinados mostraron resultados positivos a VIA y Salmonella spp., respectivamente. Además, se detectaron anticuerpos frente a T. gondii en el 9,2% de las 142 palomas y en el 26,9% de los 108 animales de zoológico testados. Estos resultados sugieren que las palomas y los animales de zoológico podrían ser empleados como especies centinela en la vigilancia de patógenos zoonósicos en zonas urbanas. En el Capítulo 2.2 se describe el primer caso mortal de infección por Acinetobacter baumannii en visón europeo (Mustela lutreola), una especie catalogada como en peligro crítico de extinción. La bacteria se aisló en cultivo de pulmón y riñón, se identificó mediante una batería de pruebas bioquímicas (API 20NE) y se confirmó mediante desorción/ionización láser asistida por matriz (MALDI-TOF). Además, la cepa aislada presentó resistencia a un total de 19 antibióticos. La confirmación de este caso no sólo es de interés para la conservación, sino también para la Salud Pública, dado que A. baumannii es uno de los patógenos más importantes implicados en las infecciones nosocomiales en humanos. El Capítulo 3 se centra en el análisis de la distribución espacio-temporal y los factores de riesgo implicados en los brotes del virus de West Nile (VWN) en caballos de España entre 2010 y 2016. Los resultados obtenidos confirmaron que el VWN ha circulado de forma endémica en España desde el año 2010. Sin embargo, su distribución no es homogénea, estando la mayoría de los brotes (92,7%) concentrados en el suroeste del país, donde se han identificaron clústeres espaciales significativos en años no consecutivos. La temperatura media anual (49,5%), la presencia de Culex pipiens (19,5%), la precipitación media anual (16,1%) y la distancia a los humedales Ramsar (14,9%), fueron las variables más asociadas con la presencia de brotes de VWN en España. Este estudio proporcionó una información valiosa para el desarrollo de programas de vigilancia basados en riesgo, orientados a una mejor prevención y control del VWN en España. En el Capítulo 4.1 se evalúa la seroprevalencia y los factores riesgo asociados a flavivirus antigénicamente relacionados (VWN, virus de Usutu y virus de la encefalitis transmitida por garrapatas (ETG)) en perros de España. La prevalencia global de anticuerpos frente a flavivirus fue del 4,8% (39/815), y se observó una seropositividad significativamente mayor en los perros de caza en comparación con los perros de compañía. Este es el primer estudio seroepidemiológico de VWN y ETG en perros en España, así como la primera descripción de circulación de ETG en el país. Estos resultados sugieren que la vigilancia serológica en perros podría ser una herramienta complementaria para monitorizar la actividad de los flavivirus emergentes en España. El objetivo del Capítulo 4.2 fue determinar la seroprevalencia y los factores de riesgo asociados a la infección por T. gondii en perros de Andalucía y Ceuta. Se detectaron anticuerpos frente a T. gondii en el 30,6% (235/769) de los perros analizados. La edad (perros adultos), la actividad (perros de caza) y el tamaño (perros grandes y medianos) fueron los principales factores de riesgo asociados a la infección por T. gondii. Los resultados indican que T. gondii está muy extendido en perros de la España peninsular y de Ceuta, lo que podría tener importantes implicaciones para la Salud Pública. En esta Tesis Doctoral se aportan nuevos conocimientos sobre la implicación de las especies domésticas y silvestres en la epidemiología de diferentes enfermedades zoonósicas de importancia médica y sanitaria. Los resultados obtenidos permitirán mejorar la vigilancia epidemiológica de enfermedades zoonósicas compartidas entre especies animales que habitan en el entorno urbano, periurbano y en el medio natural.The One Health concept (one unique health) aims to promote, improve and defend the health and wellness status of all species through cooperation and collaboration among professionals from Public, Animal and Environmental Health. This concept is undergoing an important rebirth in recent decades due to the growing interest about the epidemiological relationships of diseases shared among humans, domestic animals, wildlife and the environment, and the evidence that the control against these diseases must be established from a holistic perspective. The general objective of this Doctoral Thesis is to assess the role of different domestic, peridomestic and wild species in the epidemiology of bacterial, viral and parasitic zoonotic diseases under a One Health approach. In the first study (Chapter 1.1), we aimed to establish the seroprevalence, risk factors, spatial distribution and spoligotypes of Mycobacterium tuberculosis complex (MTC) circulating in pigs extensively farmed in Andalusia, in order to assess their role in the maintenance and transmission of tuberculosis (TB). The individual seroprevalence of MTC was 2.3% (82/3,622) and the herd prevalence was 24.8% (32/129). Pig census and the presence of neighboring goat flocks were risk factors associated with MTC seropositivity in farms. Two statistically significant spatial clusters (P<0.001) were identified and a total of 25 different MTC spoligotypes shared with other domestic and wild species were isolated in pigs bred extensively in the study area. The low individual seroprevalence found suggests that Iberian pigs could act as spillover hosts of MTC. However, the high herd prevalence, as well as the identification of significant spatial clusters, indicate the need to implement control programs in pig farms managed under extensive production systems in Spain. The aim of the Chapter 1.2 was to assess the usefulness of the proper disposal of hunting waste (big game) in high TB prevalence areas as MTC control measure in wild boar (Sus scrofa) and red deer (Cervus elaphus) (which are considered natural MTC reservoirs in Spain), from 2008/2009 to 2016/2017 hunting seasons. The results evidenced that the proper disposal of hunting waste contributed to achieve a 25% reduction in MTC seroprevalence in wild boar. This measure together with other wildlife disease control tools, should be implemented and maintained over time in integrated disease control strategies. The objective of the Chapter 2.1 was to determine the prevalence of pathogenic zoonotic agents (flaviviruses, avian influenza virus (AIV), Salmonella spp. and Toxoplasma gondii) in feral pigeons and sympatric zoo animals from Córdoba (southern Spain) between 2013 and 2014. Antibodies against flaviviruses were detected in 7.8% out of 142 pigeons, and 8.2% out of 49 zoo animals tested. Even though seropositivity to AIV and Salmonella spp. was not detected in pigeons, 17.9% (5/28) and 6.8% (3/44) of the zoo animals tested, showed positive results to AIV and Salmonella spp, respectively. Moreover, antibodies against Toxoplasma gondii were found in 9.2% out of 142 pigeons and 26.9% out of 108 zoo animals tested. These results suggest that pigeons and sympatric zoo animals may be included as sentinel species for monitoring zoonotic pathogens in urban areas. In Chapter 2.2, the first fatal case of Acinetobacter baumannii infection is reported in the critically endangered European mink (Mustela lutreola). This pathogen was isolated from lung and kidney culture, then identified through a battery of biochemical tests (API 20NE) and finally confirmed using desorption/ionization laser assisted by matrix (MALDITOF). The isolated strain showed resistance against 19 different antibiotics. Confirmation of this case is not only of conservation interest but also of public health concern given that A. baumannii is one of the most important pathogens implicated in nosocomial infections in humans. Chapter 3 analyzed the spatio-temporal distribution and risk factors involved in the West Nile virus (WNV) outbreaks in horses in Spain between 2010 and 2016. WNV has circulated endemically in Spain since 2010. However, its distribution is not homogeneous, since most of the WNV outbreaks in Spain (92.7%) were concentrated in the southwestern part of the country, where significant clusters were detected in non-consecutive years. Mean annual temperature (49.5%), presence of Culex pipiens (19.5%), mean annual precipitation (16.1%) and distance to Ramsar wetlands (14.9%) were the environmental variables most associated with WNV outbreaks occurrence in Spain. This study provided valuable information for the development of surveillance programs based on risk factors and aimed at improving prevention and control of WNV in Spain. In Chapter 4.1, the seroprevalence and risk factors associated with antigenically related flaviviruses, (WNV, Usutu virus (USUV) and tick-borne encephalitis virus (TBEV)), were assessed in dogs in Spain. The overall prevalence of antibodies against flaviviruses was 4.8% (39/815) and a significantly higher seropositivity was observed in hunting dogs compared to pet dogs. This is the first seroepidemiological survey of WNV and TBEV in dogs in Spain, as well as the first report of TBEV circulation in this country. These results suggest that serosurveillance in dogs could be a complementary tool for monitoring the activity of emerging flaviviruses in Spain. The aim of the Chapter 4.2 was to determine the seroprevalence and risk factors associated with T. gondii infection in dogs from Andalusia and Ceuta in Spain. Antibodies to T. gondii were detected in 30.6% (235/769) of the dogs analysed. The main risk factors for T. gondii infection were age (higher seroprevalence in older dogs), activity (hunting dogs) and size (higher seroprevalence in larger and medium dogs). The results indicate that T. gondii is widespread in dogs in mainland Spain and Ceuta, which might have important implications for Public health. This Doctoral Thesis contributes to throw new knowledge about the role of domestic and wild species in the epidemiology of different zoonotic diseases of medical and health concern. The results obtained will allow to improve the epidemiological surveillance of zoonotic diseases shared among animal species from urban, periurban and natural habitats

Epidemiological surveillance of Schmallenberg virus in small ruminants in southern Spain

Schmallenberg virus (SBV) is an emerging Culicoides‐borne Orthobunyavirus that affects ruminant species. Between 2011 and 2013, it was responsible for a large‐scale epidemic in Europe. In the present study, we aimed to determine the seroprevalence, spatial distribution and risk factors associated with SBV exposure in sheep and goats in the region where the first Schmallenberg disease outbreak in Spain was reported. Blood samples from 1,796 small ruminants from 120 farms were collected in Andalusia (southern Spain) between 2015 and 2017. Antibodies against SBV were detected in 536 of 1,796 animals (29.8%; 95%CI: 27.7–32.0) using a commercial blocking ELISA. The individual seroprevalence according to species was 31.1% (280/900; 95%CI: 28.1–34.1) in sheep and 28.6% (256/896; 95%CI: 25.6–31.5) in goats. The farm prevalence was 76.7% (95%CI: 69.1–84.2). Seropositivity to SBV was confirmed in both sheep and goats in all provinces by virus neutralization test. Two significant (p 14ºC) and altitude (<400 metres above sea level) were risk factors associated with SBV exposure in small ruminants. Our results highlight widespread but not homogeneous circulation of SBV in small ruminant populations in Spain.info:eu-repo/semantics/acceptedVersio

Spatiotemporal monitoring of selected pathogens in Iberian ibex (Capra pyrenaica)

An epidemiological surveillance programme was carried out to assess exposure and spatiotemporal patterns of selected pathogens (Brucella spp., Mycobacterium avium subsp. paratuberculosis (MAP), Mycoplasma agalactiae, Pestivirus and bluetongue virus (BTV)) in Iberian ibex (Capra pyrenaica) from Andalusia (southern Spain), the region with the largest population of this species. A total of 602 animals in five distribution areas were sampled during 2010–2012 (P1) and 2013–2015 (P2). The Rose Bengal test (RBT) and complement fixation test (CFT) were used in parallel to detect anti‐Brucella spp. antibodies. Commercial ELISAs were used to test for antibodies against the other selected pathogens. Sera positive for BTV and Pestivirus by ELISA were tested by serum neutralization test (SNT) to identify circulating serotypes/genotypes. The overall seroprevalences were as follows: 0.4% for Brucella spp. (2/549; CI 95%: 0.1–1.3) (14/555 positive by RBT; 2/564 by CFT), 0.5% for MAP (3/564; CI 95%: 0.1–1.5), 5.7% for M. agalactiae (30/529; CI 95%: 3.9–8.0), 11.1% for Pestivirus (58/525; CI 95%: 8.5–14.1) and 3.3% for BTV (18/538; CI 95%: 2.0–5.2). Significantly higher seropositivity to both M. agalactiae and BTV was observed in P1 compared with P2. Spatiotemporal clusters of high seroprevalence were also found for M. agalactiae in four of the five sampling areas in 2010, and for BTV in one of five areas in 2012. Specific antibodies against BTV‐4, BDV‐4 and BVDV‐1 were confirmed by SNT. Our results indicate that the Iberian ibex may be considered spillover hosts of Brucella spp. and MAP rather than true reservoirs. The prevalence of antibodies against M. agalactiae and BTV suggests spatiotemporal variation in the circulation of these pathogens, while Pestivirus has a moderately endemic circulation in Iberian ibex populations. Our study highlights the importance of long‐term surveillance for a better understanding of the spatiotemporal distribution of shared infectious diseases and providing valuable information to improve control measures at the wildlife–livestock interface.info:eu-repo/semantics/acceptedVersio

High exposure of West Nile virus in equid and wild bird populations in Spain following the epidemic outbreak in 2020

A cross-sectional study was conducted to assess the circulation and risk factors associated with West Nile virus (WNV) exposure in equine and wild bird populations following the largest epidemic outbreak ever reported in Spain. A total of 305 equids and 171 wild birds were sampled between November 2020 and June 2021. IgG antibodies against flaviviruses were detected by blocking enzyme-linked immunosorbent assay (bELISA) in 44.9% (109/243) and 87.1% (54/62) of unvaccinated and vaccinated equids, respectively. The individual seroprevalence in unvaccinated individuals (calculated on animals seropositive by both bELISA and virus microneutralization test [VNT]) was 38.3% (95%CI: 33.1–43.4). No IgM antibodies were detected in animals tested (0/243; 0.0%; 95%CI: 0.0–1.5) by capture-ELISA. The main risk factors associated with WNV exposure in equids were age (adult and geriatric), breed (crossbred) and the absence of a disinsection programme on the facilities. In wild birds, IgG antibodies against flaviviruses were found in 32.7% (56/171; 95%CI: 26.8–38.6) using bELISA, giving an individual WNV seroprevalence of 19.3% (95%CI: 14.3–24.3) after VNT. Seropositivity was found in 37.8% of the 37 species analysed. Species group (raptors), age (>1-year old) and size (large) were the main risk factors related to WNV seropositivity in wild birds. Our results indicate high exposure and widespread distribution of WNV in equid and wild bird populations in Spain after the epidemic outbreak in 2020. The present study highlights the need to continue and improve active surveillance programmes for the detection of WNV in Spain, particularly in those areas at greatest risk of virus circulation

Monitoring of bluetongue virus in zoo animals in Spain, 2007–2019

Bluetongue (BT) is an emerging and re-emerging communicable vector-borne disease of animal health concern. A serosurvey was performed to assess exposure to BT virus (BTV) in zoo animals in Spain and to determine the dynamics of seropositivity in longitudinally sampled individuals during the study period. Serum samples were collected from 241 zoo animals belonging to 71 different species in five urban zoos (A-E) in Spain between 2007 and 2019. Twenty-four of these animals were longitudinally surveyed at three of the sampled zoos (zoos B, C and E) during the study period. Anti-BTV antibodies were found in 46 (19.1%; 95% CI: 14.1-24.1) of the 241 captive animals analysed by commercial ELISA. A virus neutralization test confirmed specific antibodies against BTV-1 and BTV-4 in 25 (10.7%; 95% CI: 6.7-14.6) and five (3.0%; 95% CI: 0.3-4.0) animals, respectively. Two of the 24 longitudinally sampled individuals (one African elephant (Loxodanta africana) and one aoudad (Ammotragus lervia)) showed anti-BTV antibodies at all samplings, whereas seroconversions were detected in one mouflon (Ovis aries musimon) in 2016, and one Asian elephant (Elephas maximus) in 2019. To the best of the authors' knowledge, this is the first large-scale survey on BTV conducted in both artiodactyl and non-artiodactyl zoo species worldwide. The results confirm BTV exposure in urban zoo parks in Spain, which could be of animal health and conservation concern. Circulation of BTV was detected in yearling animals in years when there were no reports of BTV outbreaks in livestock. Surveillance in artiodactyl and non-artiodactyl zoo species could be a valuable tool for epidemiological monitoring of BTV.info:eu-repo/semantics/acceptedVersio

Epidemiological survey and risk factors associated with hepatitis E virus in small ruminants in southern Spain

Autochthonous cases of hepatitis E (HE) associated with zoonotic genotypes HEV-3 and HEV-4 have significantly increased in industrialized countries over the last decade. Suidae are generally recognized as the main reservoirs of these genotypes. Susceptibility to HE virus (HEV) infection and zoonotic potential have also been confirmed in other species, including sheep and goat. However, the information about their role in the epidemiology of HEV remains very scarce. The objective of this study was to assess the prevalence, spatial distribution and risk factors associated with HEV exposure in sheep and goats in southern Spain, the country with the highest census of small domestic ruminants in the European Union. Blood samples from 240 sheep and 240 goats were collected between 2015 and 2017. Sera were analysed in parallel using a commercial double-antigen ELISA and real-time PCR. A total of 38 (7.9%; 95%CI: 5.5–10.3) out of 480 sampled animals showed anti-HEV antibodies. By species, the seroprevalences found in sheep and goats were 2.1% (5/240; 95%CI: 0.3–3.9) and 13.8% (33/240; 95%CI: 9.4–18.1) respectively. Anti-HEV antibodies were found on 19 (59.4%; 95%CI: 42.4–76.4) of the 32 sampled farms. The GEE model showed that species (goat) and number of small ruminants in the farm (≤348 animals and ≥538 animals) were risk factors potentially associated with HEV exposure in small ruminants in the study area. HEV RNA was not detected in any of the 480 (0.0%; 95%CI: 0.0–0.8) tested animals. Our results confirm that sheep and goats are naturally, but not equally exposed to HEV and indicate the widespread spatial distribution of HEV among small ruminant populations in southern Spain. Further studies are required to elucidate the role of sheep and goat in the epidemiology of HEV and their potential implications for public health

Monitoring of Schmallenberg virus in Spanish wild artiodactyls, 2006-2015

Schmallenberg disease is an emerging disease that affects domestic and wild ruminants in Europe. An epidemiological survey was carried out to assess exposure to Schmallenberg virus (SBV) in wild artiodactyls in Spain between 2006 and 2015. A total of 1751 sera from wild artiodactyls, including 1066 red deer, 304 fallow deer, 192 mouflon, 109 wild boar, 49 roe deer and 31 Spanish ibex were tested for antibodies against SBV by ELISA and confirmed by virus neutralization test. SBV was not detected between the 2006/2007 and the 2010/2011 hunting seasons. Overall seroprevalence (including samples collected between the 2011/2012 and 2014/2015 hunting seasons) was 14.6% (160/1099; 95%CI: 12.7-16.6). Mean SBV seroprevalence was 13.3±2.6% in red deer, 23.9±4.2% in fallow deer, 16.4±6.1% in mouflon and 2.8±3.1% in wild boar. No antibodies against SBV were found in roe deer or Spanish ibex. The presence of SBV RNA was confirmed in three of 255 (1.2%) spleen samples from wild ruminants analysed by rRT-PCR. In a multivariate mixed-effects logistic regression model, the main risk factors associated with SBV seroprevalence were: species (fallow deer, red deer and mouflon), age (adults) and interactions between hunting areas of more than 1000 hectares and hunting season (2012/2013, 2013/2014 and 2014/2015). The hypothesis of endemic circulation of SBV in the last few years is supported by the detection of SBV RNA in animals sampled in 2011 and 2015, as well as antibodies detected at low level in juveniles in 2012, 2013 and 2014. The results indicate that SBV circulated in wild ruminant populations in Spain during the same period when the virus was first reported in northern Europe, and at least five months before the first case was officially reported in livestock in Spain

Evaluation of two enzyme-linked immunosorbent assays for diagnosis of bluetongue virus in wild ruminants

Bluetongue (BT) is a reportable re-emerging vector-borne disease of animal health concern. Enzyme-linked immunosorbent assays (ELISA) are frequently used in BT surveillance programs in domestic ruminants, but their diagnostic accuracy has not been evaluated for wild ruminants, which can play an important role as natural reservoirs of bluetongue virus (BTV). The aim of this study was to assess two commercial ELISAs for BT diagnosis in wild ruminants using control sera of known BTV infection status and field samples. When control sera were tested, the double recognition ELISA (DR-ELISA) showed 100 % sensitivity (Se) and specificity (Sp), while the competitive ELISA (C-ELISA) had 86.4 % Se and 97.1 % Sp. Using field samples, the selected latent-class analysis model showed 95.7 % Se and 85.9 % Sp for DR-ELISA, 58.2 % Se and 95.8 % Sp for C-ELISA and 84.2 % Se for the serum neutralization test (SNT). Our results indicate that the DR-ELISA may be a useful diagnostic method to assess BTV circulation in endemic areas, while the C-ELISA should be selected when free-areas are surveyed. The discrepancy between control and field samples point out that the inclusion of field samples is required to assess the accuracy of commercial ELISAs for the serological diagnosis of BTV in wild ruminants.info:eu-repo/semantics/acceptedVersio

Seroepidemiology of tuberculosis in sheep in southern Spain

Tuberculosis (TB) is a multi-host infectious disease caused by members of the Mycobacterium tuberculosis complex (MTC). In Mediterranean ecosystems, where multiple animal hosts of TB are present, identifying the role of the different species involved in the epidemiology of TB is a key point to be able to implement proper control measures. Sheep are susceptible to MTC infection but have traditionally been considered a spillover host. However, the occurrence of outbreaks involving sheep in recent years evidences the need to better understand the role of this small ruminant species in the epidemiology of the disease. Here, we aimed to determine the seroprevalence and risk factors associated with MTC seropositivity in sheep in Andalusia (southern Spain), a region with one of the highest prevalence of MTC infection in both cattle and wild ungulates. A total of 2266 sheep from 83 flocks were tested for antibodies against MTC using an in-house indirect ELISA. Anti-MTC antibodies were detected in 16 (0.7%) of the 2266 sheep (adjusted true prevalence 0.29%, 95% posterior probability interval 0.01–1.05). Seropositivity was found in 14.5% (12/83; 95%CI: 6.9–22.0) of the sheep farms analyzed. A semi-extensive management system was identified as a risk factor associated with MTC seropositivity in sheep farms (OR = 3.7; p < 0.038; 95%CI: 1.1–12.4) in the study area. To the best of the authors' knowledge, this is the first active TB surveillance study carried out to assess MTC exposure in sheep. Our results indicate MTC circulation in sheep farms in southern Spain. However, the low individual seroprevalence obtained suggests that sheep may play a limited role in the epidemiology of TB in this region. Serosurveillance programs could be a valuable tool to detect MTC circulation in sheep in risk scenarios or target farms, in order to optimize control measures on TB animal in multi-host Mediterranean ecosystems.This study was partially funded by the Spanish Ministry of Economy and Competitiveness (MINECO) research grant (AGL2013-49159-C2-2-R). This research was also supported by CIBER -Consorcio Centro de Investigación Biomédica en Red- (CB 2021), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación and Unión Europea – NextGenerationEUD. D. Jiménez-Martín holds a PhD contract granted by Own Research Plan of the University of Córdoba. Funding for open access charge: Universidad de Córdoba / CBUA.info:eu-repo/semantics/publishedVersio