Antibody isotype epitope mapping of SARS-CoV-2 Spike RBD protein: Targets for COVID-19 symptomatology and disease control.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) still poses a challenge for biomedicine and public health. To advance the development of effective diagnostic, prognostic, and preventive interventions, our study focused on high-throughput antibody binding epitope mapping of the SARS-CoV-2 spike RBD protein by IgA, IgM and IgG antibodies in saliva and sera of different cohorts from healthy uninfected individuals to SARS-CoV-2-infected unvaccinated and vaccinated asymptomatic, recovered, nonsevere, and severe patients. Identified candidate diagnostic (455-LFRKSNLKPFERD-467), prognostic (395-VYADSFVIRGDEV-407-C-KLH, 332-ITNLCPFGEV-342-C-KLH, 352-AWNRKRI-358-C-KLH, 524-VCGPKKSTNLVKN-536-KLH), and protective (MKLLE-487-NCYFPLQSYGFQPTNGVG-504-GGGGS-446-GGNYNYLYRLFRKSNLKPFERD-467) epitopes were validated with sera from prevaccine and postvaccine cohorts. The results identified neutralizing epitopes and support that antibody recognition of linear B-cell epitopes in RBD protein is associated with antibody isotype and disease symptomatology. The findings in asymptomatic individuals suggest a role for anti-RBD antibodies in the protective response against SARS-CoV-2. The possibility of translating results into diagnostic interventions for the early diagnosis of asymptomatic individuals and prognosis of disease severity provides new tools for COVID-19 surveillance and evaluation of risks in hospitalized patients. These results, together with other approaches, may contribute to the development of new vaccines for the control of COVID-19 and other coronavirus-related diseases using a quantum vaccinomics approach through the combination of protective epitopes.

The contribution of citizen science in the surveillance of wildlife and related arthropods.

Environmental and anthropogenic factors may significantly affect the diffusion of wild animals, enhancing the interface of human­wildlife interactions and driving the spread of pathogens and vector-borne diseases between animals and humans. However, in the last decade, the involvement of citizens in scientific research (the so-called citizen science approach, henceforth abbreviated as CS) provided a network of large-scale and cost-effective surveillance programmes of wildlife populations and their related arthropod species. Therefore, this review aims to illustrate different methods and tools used in CS studies, by arguing the main advantages and considering the limitations of this approach. The CS approach has proven to be an effective method for establishing density and distribution of several wild animal species, in urban, peri-urban and rural environments, as well a source of information regarding vector­host associations between arthropods and wildlife. Extensive efforts are recommended to motivate citizens to be involved in scientific projects to improve both their and our knowledge of the ecology and diseases of wildlife. Following the One Health paradigm, collaborative and multidisciplinary models for the surveillance of wildlife and related arthropod species should be further developed by harnessing the potentiality of the CS approach.

Zoonotic Enterocytozoon bieneusi genotypes in free-ranging and farmed wild ungulates in Spain.

Microsporidia comprises a diverse group of obligate, intracellular, and spore-forming parasites that infect a wide range of animals. Among them, Enterocytozoon bieneusi is the most frequently reported species in humans and other mammals and birds. Data on the epidemiology of E. bieneusi in wildlife are limited. Hence, E. bieneusi was investigated in eight wild ungulate species present in Spain (genera Ammotragus, Capra, Capreolus, Cervus, Dama, Ovis, Rupicapra, and Sus) by molecular methods. Faecal samples were collected from free-ranging (n = 1058) and farmed (n = 324) wild ungulates from five Spanish bioregions. The parasite was detected only in red deer (10.4%, 68/653) and wild boar (0.8%, 3/359). Enterocytozoon bieneusi infections were more common in farmed (19.4%, 63/324) than in wild (1.5%, 5/329) red deer. A total of 11 genotypes were identified in red deer, eight known (BEB6, BEB17, EbCar2, HLJD-V, MWC_d1, S5, Type IV, and Wildboar3) and three novel (DeerSpEb1, DeerSpEb2, and DeerSpEb3) genotypes. Mixed genotype infections were detected in 15.9% of farmed red deer. Two genotypes were identified in wild boar, a known (Wildboar3) and a novel (WildboarSpEb1) genotypes. All genotypes identified belonged to E. bieneusi zoonotic Groups 1 and 2. This study provides the most comprehensive epidemiological study of E. bieneusi in Spanish ungulates to date, representing the first evidence of the parasite in wild red deer populations worldwide. Spanish wild boars and red deer are reservoir of zoonotic genotypes of E. bieneusi and might play an underestimated role in the transmission of this microsporidian species to humans and other animals.

The fungal-related intracellular parasite Enterocytozoon bieneusi is a worldwide public health and veterinary problem. Here we demonstrated that it was present in wild boar, and wild and farmed red deer in Spain, with genotypes potentially capable of infecting humans, posing a public health risk.

Non-Invasive Method to Detect Infection with Mycobacterium tuberculosis Complex in Wild Boar by Measurement of Volatile Organic Compounds Obtained from Feces with an Electronic Nose System.

More effective methods to detect bovine tuberculosis, caused by Mycobacterium bovis, in wildlife, is of paramount importance for preventing disease spread to other wild animals, livestock, and human beings. In this study, we analyzed the volatile organic compounds emitted by fecal samples collected from free-ranging wild boar captured in Doñana National Park, Spain, with an electronic nose system based on organically-functionalized gold nanoparticles. The animals were separated by the age group for performing the analysis. Adult (>24 months) and sub-adult (12-24 months) animals were anesthetized before sample collection, whereas the juvenile (<12 months) animals were manually restrained while collecting the sample. Good accuracy was obtained for the adult and sub-adult classification models: 100% during the training phase and 88.9% during the testing phase for the adult animals, and 100% during both the training and testing phase for the sub-adult animals, respectively. The results obtained could be important for the further development of a non-invasive and less expensive detection method of bovine tuberculosis in wildlife populations.

Inbreeding shapes tuberculosis progression in female adult badgers (Meles meles).

In Focus: Beton, C. H., Delahay, R. J., Smith, F. A. P., Robertson, A., McDonald, R. A., Young, A. J., Burke T. A., & Hodgson, D. (2018). Inbreeding intensifies sex- and age-dependent disease in a wild mammal. Journal of Animal Ecology, 87, 1497-1499. https://doi.org/10.1111/1365-2656.12878 Increasing crossbreeding of relatives promotes inbreeding which, in turn, can cause a reduction in fitness and the emergence of a phenomenon known as inbreeding depression. Benton et al., used the badger (Meles meles)-Mycobacterium bovis model system to study the relationship between inbreeding, disease, ageing and sex. A link between multilocus homozygosity (general effect) and TB progression (measured as antibody response to infection) and between homozygosity at specific loci (local effect) and TB progression were established in this study, highlighting the importance of host genetic background in determining host disease outcomes, and thus on shaping disease dynamics in wild populations. Moreover, the authors demonstrated for the first time in a natural population that this positive association might be modulated by sex and age. In infected individuals, inbreeding only affects disease progression in adult females.

Heat-inactivated Mycobacterium bovis protects zebrafish against mycobacteriosis.

Control of mycobacterial infection constitutes a priority for human and animal health worldwide. However, effective vaccines are needed for the control of human and animal tuberculosis (TB). Adult zebrafish have become a useful model for studying the pathophysiology of mycobacterial infection and for the development of novel interventions for TB control and prevention. Recently, parenteral and oral immunization with the heat-inactivated Mycobacterium bovis vaccine (M. bovis IV) protected wild boar against TB. The objectives of this study were to provide additional support for the role of M. bovis IV in TB control using the zebrafish model and to conduct the first trial with this vaccine for the control of fish mycobacteriosis. The results showed that M. bovis IV protected zebrafish against mycobacteriosis caused by low and high infection doses of Mycobacterium marinum and provided evidence suggesting that the protective mechanism elicited by M. bovis IV in zebrafish as in other species is based on the activation of the innate immune response through the C3 pathway, with a role for the regulatory protein Akr2 in this process. These results encourage the use of M. bovis IV for TB control in different species.

Tuberculosis-Associated Death among Adult Wild Boars, Spain, 2009-2014.

We investigated adult Eurasian wild boar (Sus scrofa) survival and death in 2 tuberculosis-endemic populations with different harvest pressure in Spain. Overall, tuberculosis accounted for 30% of total deaths. Increased survival in protected areas has direct implications for wild boar management and tuberculosis control.

Evidence of shared bovine viral diarrhea infections between red deer and extensively raised cattle in south-central Spain.

BACKGROUND: Bovine viral diarrhea virus (BVDV) is a pestivirus that affects cattle production worldwide and that can infect other ungulates such as cervids and even wild boar (Sus scrofa). It is believed that domestic livestock can become infected through contact with wild animals, though it is known that infection can spread among wild animals in the absence of contact with livestock. Little is known about the sharing of BVDV infection between wild and domestic animals in the same habitat, which is important for designing eradication campaigns and preventing outbreaks, especially on hunting estates with high animal densities. RESULTS: We assessed the sharing of BVDV infections among hunted red deer, wild boar and cattle in south-central Spain. Sampled red deer (Cervus elaphus; n = 267) and wild boar (n = 52) were located on 19 hunting estates, and cattle (n = 180) were located on 18 nearby farms. We used ELISA kits for the serological screening, Taqman RT-PCR assay for the virus determination, and subsequent phylogenetic analysis for 17 RT-PCR positive sample amplicons. Fifty-two red deer (19.5%) and 82 cattle (45.6%) samples tested positive by ELISA. A high apparent prevalence (22.47%) was obtained for red deer, while only five cattle farms tested positive by RT-PCR. Conversely, no wild boar tested positive by both ELISA or RT-PCR. Eleven red deer (4.1%) tested positive by both ELISA and RT-PCR; these animals may have been sampled during the last phase of viremia, or they may represent previously exposed individuals infected by a different BVDV strain. The amplicons shared 92.7-100% identity and fell within the BVDV subgroup 1b, although nine of these (from four red deer and five cattle pools) formed a separate branch. This suggests that there might be a common BVDV infecting both cattle and red deer. Higher red deer abundance was significantly associated with greater risk that extensively raised cattle would test positive for BVDV by ELISA. CONCLUSIONS: Our findings suggest that BVDV is circulating between cattle and red deer populations in proximity, but further work is required to determine whether they share the same strain(s). These results suggest the potential of BVDV to serve as a surveillance marker in these shared habitats. High seroprevalence of BVDV in red deer from our study area suggests that although BVDV infection is common, animals usually survive the infection. Further research is needed to verify and investigate the role of red deer as a BVDV reservoir.

Spatiotemporal interactions between wild boar and cattle: implications for cross-species disease transmission.

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.

Effect of topical application of melatonin to the gingiva on salivary osteoprotegerin, RANKL and melatonin levels in patients with diabetes and periodontal disease.

This cross-section study was designed to assess the effect of topical application of melatonin to the gingiva on salivary RANKL, osteoprotegrin (OPG) and melatonin levels as well as plasma melatonin in 30 patients with diabetes and periodontal disease and in a control group of 30 healthy subjects. Salivary RANKL and OPG were measured by enzyme-linked immunosorbent assay and salivary and plasma melatonin by radioimmunoassay using commercial kits. Periodontograms were performed using the Florida Probe(®). Diabetic patients were treated with topical application of melatonin (1% orabase cream formula) once daily for 20 days. Patients with diabetes showed significantly higher mean levels of salivary RANKL than healthy subjects as well as significantly lower values of salivary OPG and salivary and plasma melatonin. After treatment with melatonin, there was a statistically significant decrease of the gingival index, pocket depth and salivary levels of RANKL, and a significant rise in salivary values of OPG. Changes of salivary OPG levels before and after topical melatonin treatment correlated significantly with changes in the gingival index and pocket depth. Treatment with topical melatonin was associated with an improvement in the gingival index and pocket depth, a reduction in salivary concentrations of RANKL and increase in salivary concentrations of OPG, which indicates that melatonin has a favorable effect in slowing osteoclastogenesis, improving the quality of alveolar bone and preventing the progression of periodontal disease.

Beware with the backpack! New hosts and pathogens identified for Ixodes simplex ticks collected from bats in the Iberian Peninsula.

To improve the knowledge on the role of bats in the maintenance and transmission of tick-borne pathogens, a molecular approach was used to characterize Anaplasma spp., Rickettsia spp., Coxiella burnetii, Borrelia burgdorferi s.l., piroplasmids, Hepatozoon spp., flaviviruses and nairoviruses in ticks collected from Iberian bats. A total of 732 bats from 25 species were captured at 38 sampling sites distributed in seven provinces of Spain between 2018 and 2022. Seventy-nine Ixodes simplex ticks were collected from 31 bats (Eptesicus isabellinus, Hypsugo savii, Myotis capaccini, Myotis emarginatus, Myotis myotis, Miniopterus schreibersii, Pipistrellus pipistrellus and Rhinolophus ferrumequinum). Sixty of 79 I. simplex were positive for at least one pathogen tested and were collected from 23 bats captured in southeast Spain. We detected the presence of Rickettsia slovaca in 12 ticks collected from M. emarginatus, H. savii, M. schreibersii and E. isabellinus; Rickettsia aeschlimannii in 1 tick from M. schreibersii; Anaplasma ovis in 3 ticks from H. savii and M. schreibersii; C. burnetii in 2 ticks from H. savii; Occidentia massiliensis in 1 tick from H. savii; piroplasmids in 12 ticks from H. savii, M. schreibersii and E. isabellinus; and a novel nairovirus in 1 tick from M. schreibersii. Furthermore, blood samples obtained from 14 of the 31 tick-infested bats were negative in all PCR analyses. This study describes new host and pathogen associations for the bat-specialist I. simplex, highlights the risk of spread of these pathogens, and encourages further research to understand the role of Iberian bats in the epidemiology of tick-borne pathogens.

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.

Trap-effectiveness and response to tiletamine-zolazepam and medetomidine anaesthesia in Eurasian wild boar captured with cage and corral traps.

BACKGROUND: Capture, handling and chemical restraint are basic techniques often needed for research or management purposes. The aim of this study was testing a combination of tiletamine-zolazepam (TZ) (3 mg/kg) and medetomidine (M) (0.05 mg/kg) on Eurasian wild boar (Sus scrofa). A total of 77 free-ranging wild boar were captured by means of portable cages and corral traps and then anaesthetized with intramuscular darts using a blowpipe. The individual response to chemical immobilization was characterized using anaesthetic, clinical, and serum biochemical variables. After the procedure, 14 of these wild boar were monitored for 20 days using GPS-GSM collars. RESULTS: Pre-release mortality during capture and handling (6.5%) was associated with severe trauma in corral traps. Capture specificity for wild boar was 96.3% and trapping effort was 16.5 days per captured wild boar. Mean induction period was 4.5 ± 2.2 min, hypnosis period enabling effective handling was 61.6 ± 25.4 min, and recovery period was 12.8 ± 12.1 min. No heart or respiratory failure due to added stress occurred and post-release monitoring by GPS-devices revealed no mortality due to anaesthesia. According to the best statistical model obtained, the main factor driving anaesthetic efficacy and stress indicators is trap type. CONCLUSIONS: Both cage and corral traps are efficient methods to capture wild boar. Cage traps are safer, as demonstrated by mortality rates as well as anaesthetic, physiological, and serum biochemical responses. This anaesthetic protocol is useful for prolonged handling of wild boar and allows sampling and collecting data for ecological and epidemiological studies.

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.

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.

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.

Occurrence and limited zoonotic potential of Cryptosporidium spp., Giardia duodenalis, and Balantioides coli infections in free-ranging and farmed wild ungulates in Spain.

Little information is currently available on the occurrence and molecular diversity of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Balantioides coli in wild ungulates and the role of these host species as potential sources of environmental contamination and consequent human infections. The presence of these three pathogens was investigated in eight wild ungulate species present in Spain (genera Ammotragus, Capra, Capreolus, Cervus, Dama, Ovis, Rupicapra, and Sus) by molecular methods. Faecal samples were retrospectively collected from free-ranging (n = 1058) and farmed (n = 324) wild ungulates from the five Spanish bioregions. Overall infection rates were 3.0% (42/1382; 95% CI: 2.1-3.9%) for Cryptosporidium spp., 5.4% (74/1382; 95% CI: 4.2-6.5%) for G. duodenalis, and 0.7% (9/1382; 95% CI: 0.3-1.2%) for B. coli. Cryptosporidium infection was detected in roe deer (7.5%), wild boar (7.0%) and red deer (1.5%), and G. duodenalis in southern chamois (12.9%), mouflon (10.0%), Iberian wild goat (9.0%), roe deer (7.5%), wild boar (5.6%), fallow deer (5.2%) and red deer (3.8%). Balantioides coli was only detected in wild boar (2.5%, 9/359). Sequence analyses revealed the presence of six distinct Cryptosporidium species: C. ryanae in red deer, roe deer, and wild boar; C. parvum in red deer and wild boar; C. ubiquitum in roe deer; C. scrofarum in wild boar; C. canis in roe deer; and C. suis in red deer. Zoonotic assemblages A and B were detected in wild boar and red deer, respectively. Ungulate-adapted assemblage E was identified in mouflon, red deer, and southern chamois. Attempts to genotype samples positive for B. coli failed. Sporadic infections by canine- or swine-adapted species may be indicative of potential cross-species transmission, although spurious infections cannot be ruled out. Molecular evidence gathered is consistent with parasite mild infections and limited environmental contamination with (oo)cysts. Free-ranging wild ungulate species would not presumably play a significant role as source of human infections by these pathogens. Wild ruminants do not seem to be susceptible hosts for B. coli.

Wild boar: an increasing concern for Aujeszky's disease control in pigs?

BACKGROUND: The goal of this study was describing the temporal evolution of Aujeszky's disease virus (ADV) contact prevalence among Eurasian wild boar (Sus scrofa) populations under different management regimes and contact likelihoods with domestic pigs. Given the recent increase in wild boar abundance throughout Europe, we hypothesized that wild boar contact with ADV would remain stable in time even after significant reduction of ADV prevalence in domestic pigs. RESULTS: Sera from 1659 wild boar were collected from 2000 to 2010 within 6 areas of the Iberian Peninsula and tested for the presence of antibodies against ADV by ELISA. According to sampling date, wild boar were grouped into three time periods. ADV prevalence was compared through period both globally and by geographic area. Overall seroprevalence for the ten-year study period was 49.6 ± 2.4%. The highest seroprevalence was recorded in areas with intense wild boar management. The annual proportion of positive wild boar sampling sites remained stable through the study period, while the percentage of domestic pig AD positive counties decreased from 70% in 2003 to 1.7% in 2010. CONCLUSIONS: Results presented herein confirmed our hypothesis that ADV would remain almost stable in wild boar populations. This evidences the increasing risk wild boar pose in the final stages of ADV eradication in pigs and for wildlife conservation.