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West Nile virus (WNV) is the most widely distributed mosquito-borne flavivirus in the world. This flavivirus can infect humans causing in some cases a fatal neurological disease and birds are the main reservoir hosts. WNV is endemic in Spain, and human cases have been reported since 2004. Although different studies analyse how climatic conditions can affect the dynamics of WNV infection, very few use long-term datasets. Between 2003 and 2020 a total of 2,724 serum samples from 1,707 common coots (Fulica atra) were analysed for the presence of WNV-specific antibodies. Mean (SD) annual seroprevalence was 24.67% (0.28) but showed high year-to-year variations ranging from 5.06% (0.17) to 68.89% (0.29). Significant positive correlations (p < 0.01) were observed between seroprevalence and maximum winter temperature and mean spring temperature. The unprecedented WNV outbreak in humans in the south of Spain in 2020 was preceded by a prolonged period of escalating WNV local circulation. Given current global and local climatic trends, WNV circulation is expected to increase in the next decades. This underscores the necessity of implementing One Health approaches to reduce the risk of future WNV outbreaks in humans. Our results suggest that higher winter and spring temperatures may be used as an early warning signal of more intense WNV circulation among wildlife in Spain, and consequently highlight the need of more intense vector control and surveillance in human inhabited areas.

A new regulation has led to the prohibition of recreational hunting on estates located within Spanish National Parks (NPs). Before the ban, eleven NPs in Spain had already reported negative ecological consequences associated with high densities of wild ungulates. The new situation that has occurred after the ban signifies that policies with which to control populations of wild ungulates in NPs, most of which do not have a sufficient natural capacity to regulate populations, depend exclusively on the parks' authorities. The banning of recreational hunting implies a series of social, ecological, economic and logistic challenges. The control of wild ungulate populations in NPs requires: i) the legal basis for culling; ii) social acceptance as regards removing animals and the extractive procedures employed in NPs; iii) the long-term monitoring of wild ungulates and the damages that they cause, and iv) sufficient financial and human resources. A more integrated management and policy plan is, therefore, required, which should be supported by two pillars: i) the sustainability of natural resources and the conservation of functional environments, and ii) providing society with explanations regarding the need to manage wild ungulates. In order to bridge the potential gap between these key pillars, it is important to involve stakeholders in the decision-making processes concerning wild ungulate management. The forthcoming changes in Spanish NPs provide a promising opportunity to make a substantial improvement to wild ungulate management in these protected areas. This management approach could, moreover, serve as an example and be transferred to other protected spaces.

Authors’ contributions JAB, JV contributed to the conception, design, data collection, laboratory work, data analysis, drafting and writing of the manuscript. MCL, PA, ADML contributed to design, data análisis and drafting of the manuscript. JAA, CG, FC, RCS participated in the data collection and drafting of the manuscript. All authors have read and approved the final manuscript., Controlling infectious diseases at the wildlife/livestock interface is often difficult because the ecological processes driving transmission between wildlife reservoirs and sympatric livestock populations are poorly understood. Thus, assessing how animals use their environment and how this affects interspecific interactions is an important factor in determining the local risk for disease transmission and maintenance. We used data from concurrently monitored GPS-collared domestic cattle and wild boar (Sus scrofa) to assess spatiotemporal interactions and associated implications for bovine tuberculosis (TB) transmission in a complex ecological and epidemiological system, Doñana National Park (DNP, South Spain). We found that fine-scale spatial overlap of cattle and wild boar was seasonally high in some habitats. In general, spatial interactions between the two species were highest in the marsh-shrub ecotone and at permanent water sources, whereas shrub-woodlands and seasonal grass-marshlands were areas with lower predicted relative interactions. Wild boar and cattle generally used different resources during winter and spring in DNP. Conversely, limited differences in resource selection during summer and autumn, when food and water availability were limiting, resulted in negligible spatial segregation and thus probably high encounter rates. The spatial gradient in potential overlap between the two species across DNP corresponded well with the spatial variation in the observed incidence of TB in cattle and prevalence of TB in wild boar. We suggest that the marsh-shrub ecotone and permanent water sources act as important points of TB transmission in our system, particularly during summer and autumn. Targeted management actions are suggested to reduce potential interactions between cattle and wild boar in order to prevent disease transmission and design effective control strategies., Junta de Comunidades Castilla-La Mancha (PEII10-0262-7673), Unión Europea (FP7 grant 613779 WildTBVac), Ministerio de Economía y Competitividad, Universidad de Castilla La Mancha, APHAEA (219235-FP7-ERA-NET-EMIDA), Depto. de Sanidad Animal, Fac. de Veterinaria, TRUE, pub

West Nile virus (WNV) is a re-emerging zoonotic pathogen with increasing incidence in Europe, producing a recent outbreak in 2020 in Spain with 77 human cases and eight fatalities. However, the factors explaining the observed changes in the incidence of WNV in Europe are not completely understood. Longitudinal monitoring of WNV in wild animals across Europe is a useful approach to understand the eco-epidemiology of WNV in the wild and the risk of spillover into humans. However, such studies are very scarce up to now. Here, we analysed the occurrence of WNV and Usutu virus (USUV) antibodies in 2102 samples collected between 2005 and 2020 from a population of feral horses in Doñana National Park. The prevalence of WNV antibodies varied between years, with a mean seroprevalence of 8.1% (range 0%–25%) and seasonally. Climate conditions including mean minimum annual temperatures and mean rainy days per year were positively correlated with WNV seroprevalence, while the annual rainfall was negatively. We also detected the highest incidence of seroconversions in 2020 coinciding with the human outbreak in southern Spain. Usutu virus-specific antibodies were detected in the horse population since 2011. The WNV outbreak in humans was preceded by a long period of increasing circulation of WNV among horses with a very high exposure in the year of the outbreak. These results highlight the utility of One Health approaches to better understand the transmission dynamics of zoonotics pathogens.

To record the abundance, density and distribution of small-mammals in Doñana Natural Area, a protocol of capture-mark and recapture monitoring of individuals is applied using Sherman’s live-traps (https://www.semice.org/es/el-proyecto/; Flowerdew et al. 2004, Torre el al., 2018). Seven localities of Doñana (MANECORRO, MATASGORDAS, MEDIANA, MORO, LORO AND SABINAR) of different mediterranean habitats (grassland, shrubland, pine forest, juniper woodlands and oak forest) have been sampled twice per year (between May-Dec) since 2011. Each locality has been divided in plots and subplots respectively: MANECORRO3 (A, B, C, D), MATASGORDAS3 (B, C, D), MATASGORDAS4 (A), MATASGORDAS5 (A), MEDIANA2 (B, C, D), MEDIANA3 (A, C, D), MORO1 (B, D), MORO2 (A, C), LORO1 (A, B, D), LORO2 (A, B, C), LORO4 (B, C, D), LORO6 (A, C, D), SABINAR1 (A), SABINAR2 (A) and SABINAR3 (A). In each subplot has been placed between 36 traps spaced 15 meters apart covering an area of 0.56 ha. The small mammals species captured have been: white-toothed shrew Crocidura russula, black rat Rattus rattus, garden dormouse Eliomys quercinus, algerian mouse Mus spretus and wood mouse Apodemus sylvaticus. Traps were installed during the morning and remained active for three consecutive nights. A daily check was made in the morning (a night check is unnecessary since Sherman traps include food, synthetic cotton inside and lined with film to withstand inclement weather). Once captured, individual was identified until species level and sexed when it was posible. Additionally, body mass of indviduals were taken using a pesola with 0.5 g accuracy. To control the recaptures individuals of R. rattus, A. sylvaticus, M. spretus, and E. quercinus were marked with a ear tag. On the other hand, individuals of E. quercinus also was marked with transponder. Individuals of C. russula were marked with hair cut or due to its high metabolism and risk of death was released without performing the procedures at moment. All animals we

Reports of West Nile virus (WNV) associated disease in humans were scarce in Spain until summer 2020, when 77 cases were reported, eight fatal. Most cases occurred next to the Guadalquivir River in the Sevillian villages of Puebla del Río and Coria del Río. Detection of WNV disease in humans was preceded by a large increase in the abundance of Culex perexiguus in the neighbourhood of the villages where most human cases occurred. The first WNV infected mosquitoes were captured approximately one month before the detection of the first human cases. Overall, 33 positive pools of Cx. perexiguus and one pool of Culex pipiens were found. Serology of wild birds confirmed WNV circulation inside the affected villages, that transmission to humans also occurred in urban settings and suggests that virus circulation was geographically more widespread than disease cases in humans or horses may indicate. A high prevalence of antibodies was detected in blackbirds (Turdus merula) suggesting that this species played an important role in the amplification of WNV in urban areas. Culex perexiguus was the main vector of WNV among birds in natural and agricultural areas, while its role in urban areas needs to be investigated in more detail. Culex pipiens may have played some role as bridge vector of WNV between birds and humans once the enzootic transmission cycle driven by Cx. perexiguus occurred inside the villages. Surveillance of virus in mosquitoes has the potential to detect WNV well in advance of the first human cases.

Competition, predation, and diseases are key factors shaping animal communities. In recent decades, lagomorphs in Europe have been impacted by virus-borne diseases that have caused substantial declines in their populations and, subsequently, in many of their predators. We examined activity and habitat-use patterns of sympatric European rabbits (Oryctolagus cuniculus L.) and Iberian hares (Lepus granatensis R.) in Doñana National Park, Spain, (DNP) during two periods of disease outbreak. In the first period (1984–1985), fecal pellet counts and roadside counts indicated that lagomorph species were segregated, with rabbits occurring in scrublands and hares in marshlands. Both species also occupied rush and fern belt ecotones. Roadside counts at sunrise, midday, sunset, and midnight revealed that rabbits and hares had the same activity patterns (crepuscular and nocturnal) in the zone of sympatry. During the second period (2005–2016), roadside counts showed that rabbits and hares were mainly nocturnal in scrublands and border marshlands. Hares occupied scrublands; a habitat previously occupied only by rabbits. These results are interpreted in light of the competition theory and predation pressure. The disease-caused decline of rabbits has likely favored hares that moved into scrublands, a vegetation type previously occupied exclusively by rabbits. The decline of rabbits in DNP has also caused the almost disappearance of this area of the Iberian lynx (Lynx pardinus), a rabbit specialist, thus enabling generalist predators to increase. Generalist predators have subsequently increased predation pressure on both rabbits and hares, causing them to switch to nocturnal activity.

Camera trapping is a widely used tool in wildlife research and conservation, and a plethora of makes and models of camera traps have emerged. However, insufficient attention has been paid to testing their performance, particularly under field conditions. In this study, we have comparatively tested five of the most frequently used makes of camera trap (Bushnell, KeepGuard, Ltl Acorn, Reconyx and Scoutguard) to identify the key factors behind their probability of detection (i.e. the probability that the camera successfully capturing a usable photograph of an animal passing through the field of view) and trigger speed (i.e. the time delay between the instant at which a motion is detected, and the time at which the picture is taken). We used 45 cameras (nine devices of each make) with infrared flash in a field experiment in which a continuous remote video was used in parallel (as a gold-standard) to discover the animals that entered the camera trap detection zone. The period (day/night), distance between animals and cameras, model, species, deployment height and activation sensitivity were significantly related to the probability of detection. This probability was lower during the night than during the day. There was a greater probability of detecting a given species when the cameras were set at its shoulder height. The interaction between species and the distance between the animals and the cameras significantly affected the trigger speed, meaning that the closer the animals that entered the detection zone, the higher the trigger speed, with substantial differences among species. This was probably related by movement speed. In conclusion, this study shows differences in the performance of camera trap models and settings, signifying that caution is required when making direct comparisons among results obtained in different experiments, or when designing new ones. These results provide empirical guidelines for best practices in camera trapping and highlight the relevance

El virus del Nilo Occidental (WNV) es un flavivirus zoonótico que circula entre las aves transmitido por mosquitos del género Culex, quienes también transmiten otros parásitos como el protozoo Plasmodium, causante de la malaria aviar. Las aves pueden morir o sufrir infecciones crónicas con altos costes fisiológicos que comprometen su estado de salud y determinan la dinámica de sus poblaciones. En el verano de 2020 y en el de 2021, se produjo el mayor brote de WNV en dos municipios sevillanos. A pesar de su relevancia en epidemiología, son infrecuentes los análisis sobre cómo las infecciones por múltiples patógenos afectan a los hospedadores. El objetivo de este estudio es 1) identificar el coste fisiológico de la infección por el WNV y 2) determinar si estos costes están mediados por la presencia de otros patógenos u el tipo de hábitat en el que ocurre (urbano vs rural). En un total de 104 gorriones comunes (Passer domesticus) procedentes de dos zonas urbanas (origen del brote) y dos rurales (cercanas al brote) se analizó los niveles de diferentes componentes del ciclo del glutatión: la glutatión peroxidasa (GPx), la glutatión reductasa (GR) y la superóxido dismutasa (SOD), el glutatión reducido (GSH), el glutatión oxidado (GSSG) y su ratio (GSH/GSSG) y el daño oxidativo provocado a lípidos (TBARS). Nuestros resultados ponen de manifiesto que la infección por WNV afecta la actividad enzimática aumentando la de GPx y disminuyendo la de GR. También afecta a los niveles moleculares de GSH y TBARS, además, en este último caso, con importantes diferencias entre ambientes urbanos (niveles más altos) y rurales (niveles más bajos), por lo que pondría de manifiesto el diferente impacto del virus en función del hábitat. No obstante, y contrario a nuestras hipótesis, no hemos observado interacción entre la infección por malaria y el WNV.

Long-term studies of community composition and relative abundance are key tools in wildlife management and biodiversity conservation. However, few studies of this kind are available for Mediterranean carnivores, especially in the Iberian Peninsula, a hotspot of mammal biodiversity in Europe. We used 15 years of carnivore monitoring data from the Doñana National Park, one of the most representative areas for carnivores in Iberia, to obtain population trends for the main Mediterranean carnivore species. They were positive for red fox, stable for badger and Egyptian mongoose, and negative for common genet and Iberian lynx. The importance of long-term datasets and the implications of the results for the studied species at global level are discussed, above all for species whose population trends are less well known. This is the case of the Egyptian mongoose, for which we present novel information on its long-term population trend in Europe, and of the Iberian lynx, an endangered species with a clear negative trend in this well-protected area.

Multiple infections or co-exposure to pathogens should be considered systematically in wildlife to better understand the ecology and evolution of host-pathogen relationships, so as to better determine the potential use of multiple pathogens as indicators to guide health management. We describe the pattern of co-exposure to several pathogens (i.e. simultaneous positive diagnosis to pathogens in an individual considering Mycobacterium tuberculosis complex lesions, and the presence of antibodies against Toxoplasma gondii, bluetongue virus, and hepatitis E virus) and assessed their main drivers in the wild ungulate community from Doñana National Park (red deer, fallow deer, and wild boar) for a 13-years longitudinal study. The lower-than-expected frequency of co-exposure registered in all species was consistent with non-mutually exclusive hypotheses (e.g. antagonism or disease-related mortality), which requires further investigation. The habitat generalist species (red deer and wild boar) were exposed to a greater diversity of pathogens (frequency of co-exposure around 50%) and/or risk factors than fallow deer (25.0% ± CI95% 4.9). Positive relationships between pathogens were evidenced, which may be explained by common risk factors favouring exposure. The specific combination of pathogens in individuals was mainly driven by different groups of factors (individual, environmental, stochastic, and populational), as well as its interaction, defining a complex eco-epidemiological landscape. To deepen into the main determinants and consequences of co-infections in a complex assemblage of wild hosts, and at the interface with humans and livestock, there also is needed to expand the range of pathogens and compare diverse assemblages of hosts under different environmental and management circumstances.

The goal of this report is modelling the occurrence for carnivores at the European scale and to compare the output of occurrence with observed hunting yield (HY) density models for red fox (Vulpes vulpes) and badger (Meles meles). Random Forest function was used for modelling occurrence of species. Occurrences available from the past 30 years (1990-2020), and HY data (period 2012-2021) from records submitted to ENETWILD were considered for modelling. Like previous models based on HY for ungulates, the response variable was the maximum number of carnivores hunted in that period divided by the area in km2 of the corresponding administrative unit (HY density). Models based on HY were statistically downscaled to make predictions to 10x10 km2. Occurrence data models indicated a good predictive performance for most species, showing that the model framework proposed for ungulates can also be applied for carnivores. Realistic distribution maps of carnivore species were achieved under this framework, except for those ones which are expanding their range, the golden jackal (Canis aureus), or those considered alien species, raccoon (Procyon lotor) and raccoon dog (Nyctereutes procyonoides); or those having a very limited distribution as the Iberian lynx (Lynx pardinus) or the steppe polecat (Mustela eversmanii): in those cases the obtained models were underestimating their suitability in Europe. Suitability has potential to be used as a proxy for abundance of red fox and badger. Validation of suitability on HY suggested the potential to be used as a proxy for abundance of red fox and badger but depending on each species. The calibration plots for HY models showed a good and linear predictive performance for fox and badger as well as an expected pattern of abundance of species, according to the data. However, differences in type of hunting and regulations in game carnivores between countries must be playing an important role in the patterns obtained. We conclude that (i) the fr

Author's summary The Dilution Effect Hypothesis (DEH) postulates that biodiversity can reduce disease epidemics because more diverse communities harbour a lower fraction of competent hosts, which thus reduces pathogen prevalence. Here, we tested DEH by using field information from 45 populations in SW Spain on the prevalence of four vector-borne pathogens and considered both the potential role of the vertebrate community and mosquito vectors. We determined the prevalence of Plasmodium, Haemoproteus, Leucocytozoon and antibodies for the zoonotic West Nile virus in wild house sparrows. Contrary to the predictions of DEH, our results do not support the general protective ability of biodiversity to reduce the prevalence of these four pathogens. The Dilution Effect Hypothesis (DEH) argues that greater biodiversity lowers the risk of disease and reduces the rates of pathogen transmission since more diverse communities harbour fewer competent hosts for any given pathogen, thereby reducing host exposure to the pathogen. DEH is expected to operate most intensely in vector-borne pathogens and when species-rich communities are not associated with increased host density. Overall, dilution will occur if greater species diversity leads to a lower contact rate between infected vectors and susceptible hosts, and between infected hosts and susceptible vectors. Field-based tests simultaneously analysing the prevalence of several multi-host pathogens in relation to host and vector diversity are required to validate DEH. We tested the relationship between the prevalence in house sparrows (Passer domesticus) of four vector-borne pathogens-three avian haemosporidians (including the avian malaria parasite Plasmodium and the malaria-like parasites Haemoproteus and Leucocytozoon) and West Nile virus (WNV)-and vertebrate diversity. Birds were sampled at 45 localities in SW Spain for which extensive data on vector (mosquitoes) and vertebrate communities exist. Vertebrate censuses were conduct

The uropygial gland of birds produces an oily secretion with different functions, mainly related to plumage protection. In addition, the volatile compounds of this secretion may act as chemical signals that provide information to conspecifics, but it is also possible that those compounds may further attract hematophagous insect vectors such as those responsible for avian malaria transmission. Individual characteristics such as sex and age are usually associated with variation in the composition of the uropygial secretion. Different studies have shown that mosquitoes are more attracted towards individual birds infected by avian malaria parasites. However, whether the individual infection status by these parasites may lead to differences in the composition of this secretion remains poorly known. We used Gas Chromatography-Mass Spectrometry (GC-MS) to characterise the chemical composition of the volatile lipophilic fraction of the uropygial gland secretions of wild European blackbirds and compare its composition in an urban and a forest locality according to their age, sex and infection status by blood parasites. We found differences in the composition of the secretion between age classes and also between sexes within adult birds. However, no differences were found in the chemical composition of the uropygial gland secretion of birds according to their infection status by blood-parasites and habitat type. These results suggest that haemosporidian infection does not alter the composition of the volatile fraction of uropygial gland secretions in infected birds.