Mapping high-risk areas for Mycobacterium tuberculosis complex bacteria transmission: Linking host space use and environmental contamination.

In many Mediterranean ecosystems, animal tuberculosis (TB), caused by Mycobacterium bovis, an ecovar of Mycobacterium tuberculosis complex (MTBC), is maintained by multi-host communities. It is hypothesised that interspecies transmission is mainly indirect via shared contaminated environments. Therefore, identifying spatial areas where MTBC bacteria occur and quantifying space use by susceptible hosts might help predict the spatial likelihood of transmission across the landscape. Here, we aimed to evaluate the transmission risk of MTBC in a multi-host system involving wildlife (ungulates and carnivores) and cattle (Bos taurus). We collected eighty-nine samples from natural substrates (water, soil, and mud) at 38 sampling sites in a TB endemic area within a Mediterranean agroforestry system in Portugal. These samples were analysed by real-time PCR to detect MTBC DNA. Additionally, host-specific space use intensity maps were obtained through camera-trapping covering the same sampling sites. Results evidenced that a significant proportion of samples were positive for MTBC DNA (49 %), suggesting that the contamination is widespread in the area. Moreover, they showed that the probability of MTBC occurrence in the environment was significantly influenced by topographic features (i.e., slope), although other non-significant predictor related with soil conditions (SMI: soil moisture index) incorporated the MTBC contamination model. The integration of host space use intensity maps with the spatial detection of MTBC showed that the red deer (Cervus elaphus) and wild boar (Sus scrofa) exhibited the highest percentages of high-risk areas for MTBC transmission. Furthermore, when considering the co-occurrence of multiple hosts, transmission risk analyses revealed that 26.5 % of the study area represented high-risk conditions for MTBC transmission, mainly in forest areas.

Spatiotemporal dynamics of Ixodes ricinus abundance in northern Spain.

Ixodes ricinus is the most medically relevant tick species in Europe because it transmits the pathogens that cause Lyme borreliosis and tick-borne encephalitis. Northern Spain represents the southernmost margin of its main European range and has the highest rate of Lyme borreliosis hospitalisations in the country. Currently, the environmental determinants of the spatiotemporal patterns of I. ricinus abundance remain unknown in this region and these may differ from drivers in highly favourable areas for the species in Europe. Therefore, our study aimed to understand the main factors modulating questing I. ricinus population dynamics to map abundance patterns in northern Spain. From 2012 to 2014, monthly/fortnightly samplings were conducted at 13 sites in two regions of northern Spain to estimate spatiotemporal variation in I. ricinus questing abundance. Local abundance of I. ricinus was modelled in relation to variation in local biotic and abiotic environmental conditions by constructing generalised linear mixed models with a zero-inflated negative binomial distribution for overdispersed data. The different developmental stages of I. ricinus were most active at different times of the year. Adults and nymphs showed a peak of abundance in spring, while questing larvae were more frequent in summer. The main determinants affecting the spatiotemporal abundance of the different stages were related to humidity and temperature. For adults and larvae, summer seemed to be the most influential period for their abundance, while for nymphs, winter conditions and those of the preceding months seemed to be determining factors. The highest abundances of nymphs and adults were predicted for the regions of northern Spain with the highest rate of Lyme borreliosis hospitalisations. Our models could be the basis on which to build more accurate predictive models to identify the spatiotemporal windows of greatest potential interaction between animals/humans and I. ricinus that may lead to the transmission of I. ricinus-borne pathogens.

Movement-driven modelling reveals new patterns in disease transmission networks.

Interspecific interactions are highly relevant in the potential transmission of shared pathogens in multi-host systems. In recent decades, several technologies have been developed to study pathogen transmission, such as proximity loggers, GPS tracking devices and/or camera traps. Despite the diversity of methods aimed at detecting contacts, the analysis of transmission risk is often reduced to contact rates and the probability of transmission given the contact. However, the latter process is continuous over time and unique for each contact, and is influenced by the characteristics of the contact and the pathogen's relationship with both the host and the environment. Our objective was to assess whether a more comprehensive approach, using a movement-based model which assigns a unique transmission risk to each contact by decomposing transmission into contact formation, contact duration and host characteristics, could reveal disease transmission dynamics that are not detected with more traditional approaches. The model was built from GPS-collar data from two management systems in Spain where animal tuberculosis (TB) circulates: a national park with extensively reared endemic cattle, and an area with extensive free-range pigs and cattle farms. In addition, we evaluated the effect of the GPS device fix rate on the performance of the model. Different transmission dynamics were identified between both management systems. Considering the specific conditions under which each contact occurs (i.e. whether the contact is direct or indirect, its duration, the hosts characteristics, the environmental conditions, etc.) resulted in the identification of different transmission dynamics compared to using only contact rates. We found that fix intervals greater than 30 min in the GPS tracking data resulted in missed interactions, and intervals greater than 2 h may be insufficient for epidemiological purposes. Our study shows that neglecting the conditions under which each contact occurs may result in a misidentification of the real role of each species in disease transmission. This study describes a clear and repeatable framework to study pathogen transmission from GPS data and provides further insights to understand how TB is maintained in multi-host systems in Mediterranean environments.

A methodological framework to characterize the wildlife-livestock interface: The case of wild boar in mainland Spain.

BACKGROUND: The representation of wildlife-livestock interface (WLI) at an accurate spatial resolution poses several challenges. Furthermore, there is a lack of published material providing detailed descriptions of geospatial techniques for the purpose of producing visual results that are interpretable and contrastable for epidemiological analysis. OBJECTIVES: Our aim is to develop a standardized, applicable, and scalable methodological framework for describing and characterizing the WLI across a large spatial extent. Subsequently, we aim to employ this framework to depict specific WLI based on different epidemiological scenarios determined by the abundance of wild boar (Sus scrofa) and domestic ungulates as an illustrative case, specifically focusing on mainland Spain. METHODS: To establish a methodological framework, we merged data from both wild and domestic sources into a hexagonal grid. We utilized data on wild boar hunting and the locations of pig, cattle, sheep, and goat farms in mainland Spain. New variables were derived from this combined dataset to illustrate the overlapping abundance between wild boar and domestic species. Finally, a cluster analysis of the generated variables was carried out, with the aim of distinguishing and characterizing various scenarios of the wild boar-domestic ungulate interface in mainland Spain. RESULTS: The hexagonal grid proved appropriate to represent and evaluate the WLI at fine spatial resolution over such broad extent. Despite the inability to ascribe a dominant livestock type and production system to a specific region, we were able to identify fifteen main areas of interest in terms of overlap. As for extensive livestock, normally at the highest risk of interaction with wild boar, the primary regions in Spain were those with dehesa agroecosystem and the Atlantic areas. Certain scenarios were particularly relevant in terms of risk for interaction and subsequent transmission of disease, namely, the case of extensive pig production in south western Spain (dehesa agroecosystem), which is especially concerned about the potential introduction of African Swine fever (ASF) in the Country. DISCUSSION AND CONCLUSIONS: We provide a basis for visualizing and understanding of different WLI scenarios, which is extensible to other regions and interfaces, and automatable where precise source of data from wildlife and livestock are available. This spatial statistics framework enables the utilization of high-resolution data, ensuring consistency on uniform grids. This aligns with the needs of high-resolution disease dissemination models based on wildlife behaviour. Such aspects are crucial for developing risk assessment and improving strategies for the prevention, control, and eradication of shared priority emerging diseases at national and international levels, such as ASF.

Effect of Myxoma Virus Species Jump on Iberian Hare Populations.

The myxoma virus species jump from European rabbits (Oryctolagus cuniculus) to Iberian hares (Lepus granatensis) has raised concerns. We assess the decline suffered by Iberian hare populations on the Iberian Peninsula and discuss the association between the effect of myxomatosis and the average abundance index, which we estimated by using hunting bags.

Epidemiologic Survey of Crimean-Congo Hemorrhagic Fever Virus in Suids, Spain.

We conducted a cross-sectional study in wild boar and extensively managed Iberian pig populations in a hotspot area of Crimean-Congo hemorrhagic fever virus (CCHFV) in Spain. We tested for antibodies against CCHFV by using 2 ELISAs in parallel. We assessed the presence of CCHFV RNA by means of reverse transcription quantitative PCR protocol, which detects all genotypes. A total of 113 (21.8%) of 518 suids sampled showed antibodies against CCHFV by ELISA. By species, 106 (39.7%) of 267 wild boars and 7 (2.8%) of 251 Iberian pigs analyzed were seropositive. Of the 231 Iberian pigs and 231 wild boars analyzed, none tested positive for CCHFV RNA. These findings indicate high CCHFV exposure in wild boar populations in endemic areas and confirm the susceptibility of extensively reared pigs to CCHFV, even though they may only play a limited role in the enzootic cycle.

The prohibition of recreational hunting of wild ungulates in Spanish National Parks: Challenges and opportunities.

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.

Assessing trends in population size of three unmarked species: A comparison of a multi-species N-mixture model and random encounter models.

Estimation of changes in abundances and densities is essential for the research, management, and conservation of animal populations. Recently, technological advances have facilitated the surveillance of animal populations through the adoption of passive sensors, such as camera traps (CT). Several methods, including the random encounter model (REM), have been developed for estimating densities of unmarked populations but require additional information. Hierarchical abundance models, such as the N-mixture model (NMM), can estimate abundances without performing additional fieldwork but do not explicitly estimate the area effectively sampled. This obscures the interpretation of its densities and requires its users to focus on relative measures of abundance instead. Hence, the main objective of our study is to evaluate if REM and NMM yield consistent results qualitatively. Therefore, we compare relative trends: (i) between species, (ii) between years and (iii) across years obtained from annual density/abundance estimates of three species (fox, wild boar and red deer) in central Spain monitored by a camera trapping network for five consecutive winter periods. We reveal that NMM and REM provided density estimates in the same order of magnitude for wild boar, but not for foxes and red deer. Assuming a Poisson detection process in the NMM was important to control for inflation of abundance estimates for frequently detected species. Both methods consistently ranked density/abundance across species (between species trend), but did not always agree on relative ranks of yearly estimates within a single population (between years trend), nor on its linear population trends across years (across years trend). Our results suggest that relative trends are generally consistent when the range of variability is large, but can become inconsistent when the range of variability is smaller.

Large scale spatio-temporal modelling of risk factors associated with tuberculosis exposure at the wildlife-livestock interface.

The management of animal tuberculosis (TB) is a priority for European Union animal health authorities. However, and despite all the efforts made to date, a significant part of Spain has as yet been unable to obtain the officially tuberculosis-free (OTF) status. Information regarding wildlife disease status is usually scarce, signifying that the role played by wildlife is usually ignored or poorly assessed in large-scale TB risk factor studies. The National Wildlife Health Surveillance Plan in Spain now provides information on infection rates in wildlife reservoirs at a national level, but there are limitations as regards the sample size, the spatio-temporal distribution of the samples, and the lack of homogeneity of the diagnostic techniques employed. The objective of the study described herein was, therefore, to employ a Bayesian approach with the intention of identifying the risk factors associated with four TB rates in cattle: prevalence, incidence, maintenance and persistence in Spain during the period 2014-2019. The modeling approach included highly informative spatio-temporal latent effects with which to control the limitations of the data. Variation partitioning procedures were carried out, and the pure effect of each factor was mapped in order to identify the most relevant factors associated with TB dynamics in cattle in each region. This made it possible to disclose that the movement of cattle, particularly from counties with herd incidence > 1%, was the main driver of the TB dynamics in cattle. The abundance of herds bred for bullfighting was retained in all four models, but had less weight than the movements. After accounting for farm-related factors, the TB prevalence in wild boar was retained in all the models and was significantly related to incidence, maintenance and persistence. With regard to the incidence, variation partitioning revealed that wildlife was the most explicative factor, thus suggesting that it plays a role in the introduction of the pathogen into uninfected herds, and consequently highlighting its importance in breakdowns. These results show, for the first time on a national scale, that wild ungulates play a relevant role in the spatio-temporal variability of TB in cattle, particularly as regards their disease status. Moreover, the spatial representation of the pure effect of each factor made it possible to identify which factors are driving the disease dynamics in each region, thus showing that it is a valuable tool with which to focus efforts towards achieving the OTF status.

Using species distribution modeling to generate relative abundance information in socio-politically unstable territories: Conservation of Felidae in the central-western region of Mexico.

The distribution range and population abundance of species provide fundamental information on the species-habitat relationship required for management and conservation. Abundance inherently provides more information about the ecology of species than do occurrence data. However, information on abundance is scarce for most species, mainly at large spatial scales. The objective of this work was, therefore, to provide information regarding the population status of six wild felids inhabiting territories in Mexico that are inaccessible or politically unstable. This was done using species distribution models derived from occurrence data. We used distribution data at a continental scale for the wild felids inhabiting Mexico: jaguar (Panthera onca), bobcat (Lynx rufus), ocelot (Leopardus pardalis), cougar (Puma concolor), margay (Leopardus wiedii), and jaguarundi (Herpailurus yagouaroundi) to predict environmental suitability (estimated by both Maxent and the distance to niche centroid, DNC). Suitability was then examined by relating to a capture rate-based index, in a well-monitored area in central western Mexico in order to assess their performance as proxies of relative abundance. Our results indicate that the environmental suitability patterns predicted by both algorithms were comparable. However, the strength of the relationship between the suitability and relative abundance of local populations differed across species and between algorithms, with the bobcat and DNC, respectively, having the best fit, although the relationship was not consistent in all the models. This paper presents the potential of implementing species distribution models in order to predict the relative abundance of wild felids in Mexico and offers guidance for the proper interpretation of the relationship between suitability and population abundance. The results obtained provide a robust information base on which to outline specific conservation actions and on which to examine the potential status of endangered species inhabiting remote or politically unstable territories in which on-field monitoring programs are not feasible.

The method matters. A comparative study of biologging and camera traps as data sources with which to describe wildlife habitat selection.

Habitat use is a virtually universal activity among animals and is highly relevant as regards designing wildlife management and conservation actions. This has led to the development of a great variety of methods to study it, of which resource selection functions combined with biologging-derived data (RSF) is the most widely used for this purpose. However this approach has some constraints, such as its invasiveness and high costs. Analytical approaches taking into consideration imperfect detection coupled with camera trap data (IDM) have, therefore, emerged as a non-invasive cost-effective alternative. However, despite the fact that both approaches (RSF and IDM) have been used in habitat selection studies, they should also be comparatively assessed. The objective of this work is consequently to assess them from two perspectives: explanatory and predictive. This has been done by analyzing data obtained from camera traps (60 sampling sites) and biologging (17 animals monitored: 7 red deer Cervus elaphus, 6 fallow deer Dama dama and 4 wild boar Sus scrofa) in the same periods using IDM and RSF, respectively, in Doñana National Park (southern Spain) in order to explain and predict habitat use patterns for three studied species. Our results showed discrepancies between the two approaches, as they identified different predictors as being the most relevant to determine species intensity of use, and they predicted spatial patterns of habitat use with a contrasted level of concordance, depending on species and scale. Given these results and the characteristics of each approach, we suggested that although partly comparable interpretations can be obtained with both approaches, they are not equivalent but rather complementary. The combination of data from biologging and camera traps would, therefore, appear to be suitable for the development of an analytical framework with which to describe and characterise the habitat use processes of wildlife.

Low impact of tuberculosis severity on wild boar body condition.

Body condition (BC), is a measure to assess the health status of domestic and wild animals. When food resources are abundant, a decrease in BC may indicate an increase in the energetic expenditure due to the effects of growth, reproduction, or disease. BC impoverishment is one of the most common clinical effects of diseases progressing chronically, such as animal tuberculosis (TB) caused by bacteria belonging to the Mycobacterium tuberculosis complex. The Eurasian wild boar (Sus scrofa) is the main wild TB reservoir in the Mediterranean basin. The specific aims of this work were to assess the relationship between sex, age and TB severity altogether on the BC of wild boar. For this purpose, we used the kidney fat index (KFI), to assess the impact of TB progression on the BC of 1372 hunter-harvested free-ranging wild boar in seven populations in southern Spain. Surprisingly, TB had only slight effects on wild boar BC and individuals exhibiting severe TB showed greater BC than TB-free individuals. The age (adults had greater BC than juveniles) and sex (females had greater BC than males) were the main BC determinants in wild boar. Sampling population and season explained more BC variability than individual factors, suggesting that other external factors might play an important role in the BC, and probably on the impact of the disease on this wild reservoir. The low impact of TB on wild boar BC suggests that individuals with severe TB and good BC represent potential long-term super-shedders of this pathogen.

Co-exposure to pathogens in wild ungulates from Doñana National Park, South Spain.

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.

Role of Dipolar Interactions on the Determination of the Effective Magnetic Anisotropy in Iron Oxide Nanoparticles.

Challenging magnetic hyperthermia (MH) applications of immobilized magnetic nanoparticles require detailed knowledge of the effective anisotropy constant (Keff ) to maximize heat release. Designing optimal MH experiments entails the precise determination of magnetic properties, which are, however, affected by the unavoidable concurrence of magnetic interactions in common experimental conditions. In this work, a mean-field energy barrier model (ΔE), accounting for anisotropy (EA ) and magnetic dipolar (ED ) energy, is proposed and used in combination with AC measurements to a specifically developed model system of spherical magnetic nanoparticles with well-controlled silica shells, acting as a spacer between the magnetic cores. This approach makes it possible to experimentally demonstrate the mean field dipolar interaction energy prediction with the interparticle distance, dij , ED ≈ 1/dij 3 and obtain the EA as the asymptotic limit for very large dij . In doing so, Keff uncoupled from interaction contributions is obtained for the model system (iron oxide cores with average sizes of 8.1, 10.2, and 15.3 nm) revealing to be 48, 23, and 11 kJ m-3 , respectively, close to bulk magnetite/maghemite values and independent from the specific spacing shell thicknesses selected for the study.

Wild Boar Effects on Fungal Abundance and Guilds from Sporocarp Sampling in a Boreal Forest Ecosystem.

Native wild boar (Sus scrofa) populations are expanding across Europe. This is cause for concern in some areas where overabundant populations impact natural ecosystems and adjacent agronomic systems. To better manage the potential for impacts, managers require more information about how the species may affect other organisms. For example, information regarding the effect of wild boar on soil fungi for management application is lacking. Soil fungi play a fundamental role in ecosystems, driving essential ecological functions; acting as mycorrhizal symbionts, sustaining plant nutrition and providing defense; as saprotrophs, regulating the organic matter decomposition; or as plant pathogens, regulating plant fitness and survival. During autumn (Sep-Nov) 2018, we investigated the effects of wild boar (presence/absence and rooting intensity) on the abundance (number of individuals) of fungal sporocarps and their functional guilds (symbiotic, saprotrophic and pathogenic). We selected eleven forested sites (400-500 × 150-200 m) in central Sweden; six with and five without the presence of wild boar. Within each forest, we selected one transect (200 m long), and five plots (2 × 2 m each) for sites without wild boar, and ten plots for sites with boars (five within and five outside wild boar disturbances), to determine the relationship between the intensity of rooting and the abundance of sporocarps for three fungal guilds. We found that the presence of wild boar and rooting intensity were associated with the abundance of sporocarps. Interestingly, this relationship varied depending on the fungal guild analyzed, where wild boar rooting had a positive correlation with saprophytic sporocarps and a negative correlation with symbiotic sporocarps. Pathogenic fungi, in turn, were more abundant in undisturbed plots (no rooting) but located in areas with the presence of wild boar. Our results indicate that wild boar activities can potentially regulate the abundance of fungal sporocarps, with different impacts on fungal guilds. Therefore, wild boar can affect many essential ecosystem functions driven by soil fungi in boreal forests, such as positive effects on energy rotation and in creating mineral availability to plants, which could lead to increased diversity of plants in boreal forests.

Environmental factors driving fine-scale ixodid tick abundance patterns.

Tick abundance is an essential demographic parameter to infer tick-borne pathogen transmission risks. Spatiotemporal patterns of tick abundance are heterogeneous, so its determinants at small spatial scales need to be understood to reduce their negative effects on hosts. Current knowledge of these determinants is scarce, especially in Mediterranean environments, limiting the possibilities for designing efficient tick control strategies. With the goal of unravelling tick abundance determinants and informing new tick management strategies, we estimated tick burdens on 1965 wild ungulates in Doñana National Park, Spain, annually between 2010 and 2020. Under the hypothesis of a predominant host influence on tick abundance, we modelled the burdens of Rhipicephalus annulatus, Hyalomma lusitanicum, and Ixodes ricinus with relevant predictors grouped into four factors: i) environment; ii) host population; iii) host individual; and iv) land-use. Generalized linear mixed models with a zero-inflated negative binomial distribution were built. Additionally, we analysed the differential contribution to abundance of each factor by deviance partitioning. We finally estimated the similarity in the environmental space of tick species by analysing their niche overlap with the environmental principal component analysis method. Our work hypothesis was confirmed for R. annulatus and H. lusitanicum, but we found that tick abundance at a fine spatial scale is jointly driven by multiple drivers, including all four factors considered in this study. This result points out that understanding the demography of ticks is a complex multifactorial issue, even at small spatial scales. We found no niche differences between the three tick species at the study spatial scale, thus showing similar host and environmental dependencies. Overall results identify that host aggregation areas displaying environmentally favourable traits for ticks are relevant tick and vector-borne pathogen transmission hotspots. Our findings will facilitate the design of new strategies to reduce the negative effects of tick parasitism.

Driver interactions lead changes in the distribution of imperiled terrestrial carnivores.

Biodiversity is dramatically declining worldwide due to the complex processes that include interactions among threats. Studies investigating how multiple drivers (inter)act to influence spatio-temporal shifts in species distribution ranges, which are among the most anticipated changes in biodiversity in the future, are still scarce in the scientific literature, particularly at meaningful conservation planning scales. We used species occurrence data for eight terrestrial Mediterranean carnivores at a regional scale of southern Spain to: 1) estimate environmental favourability to each species during two periods (1960s and present); 2) predict current favourability assuming the same environment-species relationships as the ones observed in the 1960s (expected favourability); and 3) analyse single and interactive effects of human persecution, presence of protected areas, and prey richness on favourability changes (present vs. expected) for each carnivore species. We found that, with a few exceptions, environmentally favourable areas for carnivore occurrence were more widely distributed in the past than today and, for most species, favourability tended to increase inside protected areas, with low frequency of poisoning events and high prey richness. Notwithstanding, for rare species such as Iberian grey wolf (Canis lupus signatus) and Iberian lynx (Lynx pardinus), interactions among drivers were the most important predictors of favourability changes, underpinned by a compensatory role of protected areas in the face of continued human persecution. We emphasize the role of protected areas in abating the effects of biodiversity threats and claim that scientific studies based on analyses of single-effect drivers that omit driver interactions may misinform conservation planning and require revision.

Survey of Culicoides-borne bluetongue and Schmallenberg viruses at the wildlife-livestock interface in Doñana National Park (Spain).

A cross-sectional study was carried out to assess the circulation of bluetongue virus (BTV) and Schmallenberg virus (SBV) within the wild and domestic ungulate host community in Doñana National Park (southwestern Spain). A total of 440 animals, including 138 cattle, 102 red deer (Cervus elaphus), 101 fallow deer (Dama dama) and 99 wild boar (Sus scrofa), were sampled in 2015 during the seasonal peak of Culicoides spp. (summer-autumn). Serum and spleen samples were analysed to detect exposure (using commercial blocking ELISAs) and infection (by RT-PCR), respectively, to BTV and SBV. Cattle were not tested by BTV-bELISA because all were previously vaccinated against BTV serotypes 1 and 4. High BTV seroprevalences were found in red deer (97.0%) and fallow deer (64.7%). Antibodies against SBV were detected in 37.0% of cattle, 16.8% of red deer, 23.5% of fallow deer and 2.0% of wild boar. Thirty-eight of the 203 deer (18.7%; 17 red deer and 21 fallow deer) were co-exposed to both viral agents. BTV-4 RNA was confirmed in four red deer and two fallow deer. SBV RNA was found in two fallow deer. Co-infections were not detected. Our results indicate high exposure, widespread distribution and active circulation of BTV and SBV in the ruminant community in the study area. We provide additional evidence for the potential role of wild cervids as reservoirs of these Culicoides-borne viruses in two different epidemiological scenarios: with vaccination (BTV) and without vaccination (SBV) of sympatric livestock. This study highlights the importance of wildlife surveillance, particularly of cervid species, for the proper execution of control programmes of Culicoides-borne diseases in extensively reared livestock.

Is serology a realistic approach for monitoring red deer tuberculosis in the field?

Tuberculosis (TB) is a zoonotic mycobacterial infection with great importance in human health, animal production, and wildlife conservation. Although an ambitious eradication programme in cattle has been implemented for decades, TB-free status has not yet been achieved in most of Spain, where animal TB persists in a multi-host system of domestic and wild hosts, including the red deer (Cervus elaphus). However, information on long time series and trends of TB prevalence in wildlife is scarce. The diagnosis of TB in wild red deer is often based on gross pathology and bacteriological culture confirmation, although recently serological assays have been developed to detect anti- Mycobacterium tuberculosis Complex (MTC) antibodies. Particularly, protein complex P22 has demonstrated to yield good specificity and sensitivity in the serological diagnosis of MTC for red deer, as well as cattle, goats, sheep, pigs, wild boar, and European badger. Thus, the objective of the present study was to compare the performance of the P22-ELISA with TB-compatible lesion detection, as well as to assess the potential application of each technique for determining spatiotemporal trends and risk factors of MTC infection in wild red deer from low and high TB prevalence areas of Spain over the last two decades. We tested 5095 sera from 13 wild populations by indirect ELISA using P22 as antigen. Mean seroprevalence (13.22%, CI95: 12.32-14.18) was compared with the prevalence of macroscopic TB-compatible lesions (6.94%, CI95: 6.18-7.79). The results evidenced a poor agreement between both techniques (K < 0.3), although generalized TB-lesions and anti-P22 antibodies showed a positive association (χ² = 9.054, P = 0.004). Consequently, TB-lesion based prevalence and seroprevalence cannot be considered as equivalent for TB surveillance in red deer. Regarding the spatiotemporal trend of TB in red deer in Spain, we observed a North-South gradient of TB occurrence [North: 1.23% (CI95: 0.77-1.97) of TB-lesions and 12.55% (CI95: 10.91-14.41) of P22-ELISA; Centre: 7.10% (CI95: 6.04-8.33) and 8.74% (CI95: 7.57-10.08); South: 21.04% (CI95:17.81-24.69) and 23.09% (CI95: 19.73-26.84), respectively]. Overall, there was a stability over time, with higher prevalence in adults belonging to densely populated sites. We conclude that the P22-ELISA alone is not sufficiently reliable for TB surveillance in red deer at large spatiotemporal scales. Instead, we recommend combining gross pathology and P22-ELISA.

Permeability of artificial barriers (fences) for wild boar (Sus scrofa) in Mediterranean mixed landscapes.

BACKGROUND: Fences are one of the most widespread manmade features in nature, constituting an artificial limitation to the movement of wildlife. To date, their effects on wildlife behavior have been understudied but this knowledge is required to design effective management procedures. Using 21 GPS-monitored wild boar, we evaluated the permeability of different types of fences and described temporal patterns and spatial hotspots for crossing events. A fence's permeability was inferred by the crossing success, i.e., the number of times that animals crossed a barrier vs the number of times they did not cross. The vulnerability of fences at watercourses was explored by assessing whether the frequency of crossings was higher around watercourse intersections than expected by chance. RESULTS: Well-maintained big game proof fences were the most effective in reducing successful wild boar crossings; they were, on average, 30% more efficient than livestock type fences. Crossing success was higher for males than females and during the food shortage period than in the food abundance period. The frequency of crossings around watercourses was higher than expected by chance, especially in moderately and well-maintained big game proof type fences. CONCLUSION: While no fence type was 100% wild boar proof, well-maintained big game proof fences substantially constrained the movement of boar. However, they are vulnerable around watercourses. Managing the conflicts in which this species is involved, such as shared infections and agricultural damage, would require fences that are even more effective than the ones analyzed here, ideally in conjunction with other preventive actions. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.