Zoo animals as sentinels for Schmallenberg virus monitoring in Spain.

Schmallenberg virus (SBV) is a newly emerged vector-borne pathogen that affects many domestic and wild animal species. A serosurvey was carried out to assess SBV exposure in zoo animals in Spain and to determine the dynamics of seropositivity in longitudinally sampled individuals. Between 2002 and 2019, sera from 278 animals belonging to 73 different species were collected from five zoos (A-E). Thirty-one of these animals were longitudinally sampled at three of these zoo parks during the study period. Seropositivity was detected in 28 (10.1 %) of 278 animals analyzed by blocking ELISA. Specific anti-SBV antibodies were confirmed in 20 (7.2 %; 95 %CI: 4.2-10.3) animals of six different species using virus neutralization test (VNT). The multiple logistic regression model showed that "order" (Artiodactyla) and "zoo provenance" (zoo B; southern Spain) were risk factors potentially associated with SBV exposure. Two (8.7 %) of the 31 longitudinally-sampled individuals showed specific antibodies against SBV at all samplings whereas seroconversion was detected in one mouflon (Ovis aries musimon) and one Asian elephant (Elephas maximus) in 2016 and 2019, respectively. To the best of the author's knowledge, this is the first surveillance conducted on SBV in zoos in Spain. The results confirm SBV exposure in zoo animals in this country and indicate circulation of the virus before the first Schmallenberg disease outbreak was reported in Spain. Surveillance in zoological parks could be a complementary approach to monitoring SBV activity. Further studies are warranted to assess the impact of this virus on the health status of susceptible zoo animals.

Evidence of exposure to zoonotic flaviviruses in zoo mammals in Spain and their potential role as sentinel species.

A serosurvey was carried out to assess emerging flavivirus exposure in zoo mammals in Spain and to determine the dynamics of seropositivity in species that were longitudinally sampled during the study period. Sera from 570 zoo animals belonging to 120 mammal species were collected at ten zoos (A-J) in Spain between 2002 and 2019. Twenty-one of these animals, belonging to ten different species, were sampled longitudinally at four of the zoos during the study period. Antigenically-related flavivirus antibodies were detected in 19 (3.3 %; 95 %CI: 2.0-5.2) of the 570 animals analyzed using bELISA. Seropositivity was observed in ten (8.3 %) of the 120 species tested. Five (23.8 %) of the 21 animals sampled more than once presented seropositivity in all samplings whereas seroconversion was only observed in one white rhinoceros (Ceratotherium simum). Flavivirus antibodies were found at six of the ten sampled zoos and in consecutive years between 2008 and 2018. Virus neutralization tests confirmed West Nile virus (WNV), Usutu virus (USUV) and tick-borne encephalitis virus (TBEV) infection in ten (1.8 %; 95 %CI: 0.7-2.8), five (0.9 %; 95 %CI: 0.1-1.6) and one (0.2 %; 95 %CI: 0.0-0.5) animal, respectively. Antibodies against Meaban virus (0 %; 95 %CI: 0.0-0.7 %) were not found in the tested sera. The results demonstrate WNV, USUV and TBEV exposure in zoo mammals, which may be of public health and conservation concern. Seropositivity to WNV and USUV was detected in regions where these viruses have not been reported previously. Anti-WNV antibodies found in zoo animals sampled in 2009 point to WNV circulation at least one year before the first outbreaks were reported in horses and humans in Spain. Our results indicate that zoo mammals could be useful sentinel species for monitoring emerging flavivirus activity in urban areas.

Survey of Aujeszky's Disease Virus in Hunting Dogs from Spain.

Direct contact with swine infected by Aujeszky's disease virus (ADV) represents a potential risk of transmission to carnivore species, in which the infection is normally fatal. We assessed exposure to ADV in hunting dogs in an area where the virus is highly endemic in wild boar populations. Two out of 466 (0.43%; 95% CI 0.00-1.02%) hunting dogs analyzed were positive by gE-bELISA, gB-bELISA and the virus neutralization test. The seroprevalence levels detected, as well as the absence of reports of clinical cases in the hunting dog groups tested, indicate limited contact of this species with ADV in the study area. Further studies are warranted to assess the pathogenicity of Aujeszky's disease virus strains infecting wild boar.

Management of hunting waste as control measure for tuberculosis in wild ungulates in south-central Spain.

In recent decades, habitat change and the intensive management of wild ungulates for hunting have led to an increase in their populations in south-central Spain. This implies a higher generation of hunting waste, which can favour the transmission of infectious diseases, including tuberculosis (TB). The aim of this study was to assess the usefulness of the proper disposal of hunting waste as TB control measure in wild boar (Sus scrofa) and red deer (Cervus elaphus) during the 2008/2009 to 2016/2017 hunting seasons. Blood samples from 664 wild boar and 934 red deer were obtained in 14 game estates in two provinces in Andalusia (Area 1), where the disposal of hunting waste was implemented since the 2012/2013 hunting season. Besides, six game estates in the province of Ciudad Real, in Castilla-La Mancha (Area 2), an adjacent region where this management measure was not implemented during the studied period, were used as controls, sampling 277 wild boar and 427 red deer sera. The Mycobacterium tuberculosis complex (MTC), seroprevalence detected in wild boar from Area 1, was significantly higher before the disposal of big game hunting by-products (82.8%; 2008/2009-2012/2013) compared to the second period (61.8%; 2013/2014-2016/2017) (p < .001), after this control measure became established. By contrast, no significant differences between periods were found in wild boar (41.3% versus 44.8%; p = .33) and red deer (14.9% versus 11.6%; p = .19) from Area 2 as well as in red deer (10.8% versus 10.5%; p = .48) from Area 1. The proper disposal of hunting waste contributed to achieve a 25% reduction in MTC seroprevalence in wild boar. These results are of particular relevance regarding wild boar in the current context of re-emerging and emerging diseases such as TB and African Swine Fever in Europe. Further studies are needed to assess the effect of this measure on the health status of livestock and other wildlife species.

Assessing the variability in transmission of bovine tuberculosis within Spanish cattle herds.

In Spain, despite years of efforts to eradicate bovine tuberculosis (bTB), the disease is still endemic, with some areas of high prevalence. In this context, the surveillance and control plans may need to be re-evaluated, and understanding the dynamics of bTB spread within Spanish herds may help to develop new strategies for reducing the time for detection of infected herds and for the elimination of bTB from the herds already infected. Here, we developed a compartmental stochastic model to simulate bTB within-herd transmission, fed it with epidemiological data from 22 herds (obtained from a previous work) and carried out parameter inference using Approximate Bayesian Computing methods We also estimated the "Within-herd transmission potential Number" (Rh), i.e. the average number of secondary cases generated by a single animal infected introduced into a totally susceptible herd, considering different scenarios depending on the frequency of controls. The median global values obtained for the transmission parameters were: for the transmission coefficient (ß), 0.014 newly infected animals per infectious individual per day (i.e. 5.2 per year), for the rate at which infected individuals become infectious (α), 0.01 per day (equivalent to a latent period of 97 days), and for the rate at which infected individuals become reactive to the skin test (α1), 0.08 per day (equivalent to a period of 12 days for an infected animal to become reactive). However, the results also evidenced a great variability in the estimates of those parameters (in particular ß and α) among the 22 herds. Considering a 6-month interval between tests, the mean Rh was 0.23, increasing to 0.82 with an interval of 1 year, and to 2.01 and 3.47 with testing intervals of 2 and 4 years, respectively.

Exposure to West Nile virus and tick-borne encephalitis virus in dogs in Spain.

In the past decade, the spread of emerging zoonotic flaviviruses (genus Flavivirus, family Flaviviridae) has been reported in many regions worldwide, representing a threat to both human and animal health. A serosurvey was carried out to assess exposure and risk factors associated with antigenically related flaviviruses, particularly West Nile virus (WNV), Usutu virus (USUV) and tick-borne encephalitis virus (TBEV), in dogs in Spain. Flavivirus antibodies were detected in 39 of 815 dogs (4.8%; 95% CI: 3.3-6.3) by bELISA. Significantly higher seropositivity was observed in hunting dogs compared to pet dogs. Virus neutralization tests confirmed WNV-specific and TBEV-specific antibodies in 11 and 14 bELISA-positive dogs, respectively. This is the first serosurvey of WNV and TBEV in dogs in Spain and the first report of TBEV circulation in this country. The seropositivity obtained indicates widespread, but not homogeneous, distribution of WNV and TBEV in dogs in Spain. In 2013 and 2015, WNV-seropositive dogs were detected in those areas of Andalusia where the highest number of WNV outbreaks were reported in both horses and humans. Antibodies against TBEV have been found in dogs sampled in two different periods and regions in Spain. Serosurveillance in dogs could be a complementary way of monitoring the activity of emerging flaviviruses in Spain.

Toxoplasma gondii in sympatric domestic and wild ungulates in the Mediterranean ecosystem.

Toxoplasma gondii is a zoonotic protozoan of worldwide distribution. The present study provides information on risk factors affecting T. gondii infection in domestic and free-ranging wild ungulates sharing habitats in Mediterranean ecosystems in Spain. Serum samples from 482 extensively reared domestic ruminants and 2351 wild ungulates were tested for T. gondii antibodies using the modified agglutination test (MAT, cut-off 1:25). Toxoplasma gondii seroprevalence was 41.2% of 194 sheep, 18.6% of 199 cattle and 5.6% of 89 goats. The main risk factors associated with infection in livestock were the presence of cats, feeding on the ground and at stubble fields. In wild ungulates, T. gondii antibodies were detected in 10.5% of 1063 red deer, 15.6% of 294 fallow deer, 5.6% of 216 European mouflon, 5.6% of 90 Spanish ibex, 13.6% of 22 roe deer and 18.6% of 666 wild boars. The risk factors affecting T. gondii infection in wildlife were species, age and hunting season. Significantly higher seroprevalence was found in domestic ruminants, particularly in sheep, compared to the wild species tested. The present study indicates widespread exposure to T. gondii among domestic and wild ungulates in Southern Spain, with significant differences among species sharing the same ecosystem. The high seroprevalence observed in domestic ruminants, particularly in sheep, reinforces the need for farm management practices to control the risk factors associated with T. gondii infection in extensively reared livestock. Consumption of raw and undercooked food products from domestic and wildlife species may have important implications for public health.

Epidemiology and spatio-temporal analysis of West Nile virus in horses in Spain between 2010 and 2016.

During the last decade, West Nile virus (WNV) outbreaks have increased sharply in both horses and human in Europe. The aims of this study were to evaluate characteristics and spatio-temporal distribution of WNV outbreaks in horses in Spain between 2010 and 2016 in order to identify the environmental variables most associated with WNV occurrence and to generate high-resolution WNV suitability maps to inform risk-based surveillance strategies in this country. Between August 2010 and November 2016, a total of 403 WNV suspected cases were investigated, of which, 177 (43.9%) were laboratory confirmed. Mean values of morbidity, mortality and case fatality rates were 7.5%, 1.6% and 21.2%, respectively. The most common clinical symptoms were as follows: tiredness/apathy, recumbency, muscular tremor, ataxia, incoordination and hyperaesthesia. The outbreaks confirmed during the last 7 years, with detection of WNV RNA lineage 1 in 2010, 2012, 2013, 2015 and 2016, suggest an endemic circulation of the virus in Spain. The spatio-temporal distribution of WNV outbreaks in Spain was not homogeneous, as most of them (92.7%) were concentrated in western part of Andalusia (southern Spain) and significant clusters were detected in this region in two non-consecutive years. These findings were supported by the results of the space-time scan statistics permutation model. A presence-only MaxEnt ecological niche model was used to generate a suitability map for WNV occurrence in Andalusia. The most important predictors selected by the Ecological Niche Modeling were as follows: mean annual temperature (49.5% contribution), presence of Culex pipiens (19.5% contribution), mean annual precipitation (16.1% contribution) and distance to Ramsar wetlands (14.9% contribution). Our results constitute an important step for understanding WNV emergence and spread in Spain and will provide valuable information for the development of more cost-effective surveillance and control programmes and improve the protection of horse and human populations in WNV-endemic areas.

Spatial-temporal Trends and Factors Associated with the Bluetongue Virus Seropositivity in Large Game Hunting Areas from Southern Spain.

An epidemiological study was carried out to determine the spatial-temporal trends and risk factors potentially involved in the seropositivity to bluetongue virus (BTV) in hunting areas with presence of red deer (Cervus elaphus). A total of 60 of 98 (61.2%; CI95% : 51.6-70.9) hunting areas sampled presented at least one seropositive red deer. Antibodies against BTV were detected in juvenile animals during the hunting seasons 2007/2008 to 2013/2014 in 15 of 98 (15.3%) hunting areas, which indicates an uninterrupted circulation of BTV in this period. A multivariate logistic regression model showed that the red deer density at hunting area level (>22 individuals/km(2) ), the annual abundance of Culicoides imicola (>1.4 mosquitoes/sampling) and the goat density at municipality level (>24.1 individuals/km(2) ) were factors significantly associated with BTV seropositivity in hunting areas. Control measures against BTV in the studied area include vaccination programmes in wild and domestic ruminants, movement control in areas with high densities and abundance of red deer and C. imicola, respectively. Considering the potential risk of BTV re-emergence, red deer should be included in the BT surveillance programmes in regions where these species share habitats with livestock.

Abdomen agudo por rotura diferida de diafragma / Acute abdomen due delayed diaphragmatic rupture

La rotura traumática del diafragma es una lesión poco frecuente que puede pasar desapercibida a menos que exista un elevado índice de sospecha. En el intervalo entre la rotura y la herniación del contenido abdominal hacia la cavidad torácica, los síntomas y signos son inespecíficos y las pruebas diagnósticas como la radiografía de tórax pueden ser normales. Presentamos un caso de manifestación tardía de una rotura diafragmática izquierda secundaria a un traumatismo torácico en accidente de tráfico. El paciente pedeció la estrangulación del segmento herniado del estómago en el tórax. Se revisa el mecanismo de la lesión y las técnicas diagnósticas de imagen (AU)