Evaluating hunting and capture methods for urban wild boar population management.

Wild ungulates are expanding in range and number worldwide leading to an urgent need to manage their populations to minimize conflicts and promote coexistence with humans. In the metropolitan area of Barcelona (MAB), wild boar is the main wildlife species causing a nuisance, from traffic accidents to health risks. Selective harvesting of specific sex and age classes and reducing anthropogenic food resources would be the most efficient approach to dealing with overpopulation. Nonetheless, there is a gap in knowledge regarding the age and sex selectivity of the capture methods currently applied in the MAB for wild boar population control. Thus, this study aimed to evaluate the performance and age and sex bias of different hunting and capture methods and the seasonal patterns in their performance (number of captured individuals per event). From February 2014 to August 2022, 1454 wild boars were captured in the MAB using drop net, teleanaesthesia, cage traps, night stalks, and drive hunting. We applied generalized linear models (GLM) to compare the performance of these methods for the total number of wild boars, the wild boars belonging to each age category (i.e., adult, yearling, and juvenile), and for each season. The studied capture methods showed age-class bias and sex bias in adults (>2 years). Drive hunting and drop net removed mainly adult females and yearlings (1-2 years), with drive hunting having the highest performance for adult males. Instead, cage traps and drop net were the best methods to capture juveniles (<1 year). Overall, global performance was higher in summer, decreasingly followed by autumn and spring, winter being the worst performing season. Wildlife managers and researchers should consider the different performance and sex and age bias of each hunting and capture method, as well as the associated public cost, to improve efficiency and achieve the best results in wild boar population management.

Between conflict and reciprocal habituation: Human-wild boar coexistence in urban areas.

Urbanization is an ongoing global environmental change. Wildlife may respond using anthropized environments and resources, which is known as synurbization, creating human-wildlife interactions. Wild boar (Sus scrofa) populations have become common in urban areas, including the metropolitan area of Barcelona. Humans respond to wild boars in urban environments either habituating, with lower conflict perception and higher wild boar acceptance, or sensitizing, with reduced tolerance towards wild boars. Since citizen response influences conflict management, this study analysed the drivers of human responses, which should allow adopting socially-accepted measures to manage synurbic wild boar populations. Interviews to 1956 Barcelona citizens were performed, grouping the response variables to score citizen and urban characteristics, as well as citizen lay-knowledge, emotions, experiences, and perception of wild boar. Five citizen clusters were identified: cluster 1 (3.3 %), highly habituated and active wild boar feeder; cluster 2 (11.3 %), habituated to wild boars with positive feelings; cluster 3 (19.8 %), not habituated nor sensitized, willing to maintain urban wild boar populations; cluster 4 (29.1 %), sensitized and concerned, defending to reduce wild boar; and cluster 5 (40.1 %), highly sensitized and proposing to reduce or even eliminate wild boar. Positive attitudes associated wild boar to aesthetic value, closeness to nature and sympathy, and were more frequent in young citizens with urban background and high education, animal lovers habituated to wild boar through contact without negative experiences. Conversely, negative attitudes were concerned about city fouling, safety or health, accepted lethal management measures, and were more frequent in older citizens with rural background, lower education, low contact with wild boar or sensitized through negative experiences. We document for the first time the sensitization and reciprocal habituation of humans to wild boar in urban areas. The identification of the drivers of citizen attitudes towards urban wildlife should be useful to design socially-accepted management measures.

Mapping the risk of exposure to Crimean-Congo haemorrhagic fever virus in the Iberian Peninsula using Eurasian wild boar (Sus scrofa) as a model.

Crimean-Congo haemorrhagic fever (CCHF) virus (CCHFV) is a tick-borne zoonotic pathogen that can cause a lethal haemorrhagic disease in humans. Although the virus appears to be endemically established in the Iberian Peninsula, CCHF is an emerging disease in Spain. Clinical signs of CCHFV infection are mainly manifested in humans, but the virus replicates in several animal species. Understanding the determinants of CCHFV exposure risk from animal models is essential to predicting high-risk exposure hotspots for public health action. With this objective in mind, we designed a cross-sectional study of Eurasian wild boar (Sus scrofa) in Spain and Portugal. The study analysed 5,291 sera collected between 2006 and 2022 from 90 wild boar populations with a specific double-antigen ELISA to estimate CCHFV serum prevalence and identify the main determinants of exposure probability. To do so, we statistically modelled exposure risk with host- and environment-related predictors and spatially projected it at a 10 × 10 km square resolution at the scale of the Iberian Peninsula to map foci of infection risk. Fifty-seven (63.3 %) of the 90 populations had at least one seropositive animal, with seroprevalence ranging from 0.0 to 88.2 %. Anti-CCHFV antibodies were found in 1,026 of 5,291 wild boar (19.4 %; 95 % confidence interval: 18.3-20.5 %), with highest exposure rates in southwestern Iberia. The most relevant predictors of virus exposure risk were wild boar abundance, local rainfall regime, shrub cover, winter air temperature and soil temperature variation. The spatial projection of the best-fit model identified high-risk foci as occurring in most of western and southwestern Iberia and identified recently confirmed risk foci in eastern Spain. The results of the study demonstrate that serological surveys of CCHFV vector hosts are a powerful, robust and highly informative tool for public health authorities to take action to prevent human cases of CCHF in enzootic and emergency settings.

The local skin cellular immune response determines the clinical outcome of sarcoptic mange in Iberian ibex (Capra pyrenaica).

Introduction: Sarcoptic mange, caused by Sarcoptes scabiei, is a disease with implications for wildlife conservation and management. Its severity depends on the host's local skin immune response, which is largely unknown in Iberian ibex (Capra pyrenaica), a mountain ungulate dramatically affected by mange. In this species, the clinical outcome of sarcoptic mange varies among individuals, and the local immune response could be key to controlling the infestation. This study aims to characterize the local cellular immune response and its relationship with the clinical outcome. Methods: Fourteen Iberian ibexes were experimentally infested with S. scabiei and six more served as controls. Clinical signs were monitored, and skin biopsies were collected from the withers at 26, 46, and 103 days post-infection (dpi). The presence and distribution of macrophages (including M1 and M2 phenotypes), T lymphocytes, B lymphocytes, plasma cells, and interleukine 10 were quantitatively evaluated using immunohistochemical techniques. Results: An inflammatory infiltrate that decreased significantly from 26 to 103 dpi was observed in all the infested ibexes. The predominant inflammatory cell population in the skin of the mangy ibexes was formed by macrophages (mainly the M2 phenotype) followed by T lymphocytes, with lower numbers of B lymphocytes and plasma cells. Three clinical courses were identified: total recovery, partial recovery, and terminal stage. The inflammatory infiltrates were less pronounced in the fully recovered ibexes than in those that progressed to the terminal stage throughout the study. Discussion: The results suggest an exacerbated but effective Th1-type cellular immune response controlling mange in Iberian ibex. Furthermore, the local immune response appears to determine the variability of the clinical responses to S. scabiei infestation in this species. This first report on the progression of local skin immune cells is relevant not only for individuals but also for population management and conservation.

Diseases of Iberian ibex (Capra pyrenaica).

Iberian ibex (Capra pyrenaica) is an ecologically and economically relevant medium-sized emblematic mountain ungulate. Diseases participate in the population dynamics of the species as a regulating agent, but can also threaten the conservation and viability of vulnerable population units. Moreover, Iberian ibex can also be a carrier or even a reservoir of pathogens shared with domestic animals and/or humans, being therefore a concern for livestock and public health. The objective of this review is to compile the currently available knowledge on (1) diseases of Iberian ibex, presented according to their relevance on the health and demography of free-ranging populations; (2) diseases subjected to heath surveillance plans; (3) other diseases reported in the species; and (4) diseases with particular relevance in captive Iberian ibex populations. The systematic review of all the information on diseases affecting the species unveils unpublished reports, scientific communications in meetings, and scientific articles, allowing the first comprehensive compilation of Iberian ibex diseases. This review identifies the gaps in knowledge regarding pathogenesis, immune response, diagnostic methods, treatment, and management of diseases in Iberian ibex, providing a base for future research. Moreover, this challenges wildlife and livestock disease and wildlife population managers to assess the priorities and policies currently implemented in Iberian ibex health surveillance and monitoring and disease management.

Tissue-specific assessment of oxidative status: Wild boar as a case study.

In recent decades, there has been a fast-growing interest in using biomarkers of oxidative stress (BOS) in conservation programs of many vertebrate species. Biomarkers of oxidative stress can be measured in different biological samples (e.g., body fluids and tissues). However, since comparisons of the same battery of BOS among tissues of the same individual are scarce in the literature, the chosen target tissues regularly rely on arbitrary decisions. Our research aimed to determine if the oxidative status of free-ranging wild boar (Sus scrofa) naturally infected with Mycobacterium spp (etiological agent of tuberculosis, TB), varies depending on the sample where it was quantified. We compared antioxidant p-nitrophenyl esterase activity (EA), glutathione peroxidase (GPX) concentrations, and total oxidative status (TOS) in serum, lung, spleen, kidney, and muscle of 63 wild boar hunter-harvested in central Spain. Biomarkers of oxidative stress in serum had higher concentrations than in other tissues. The poor agreement between serum and other tissues highlights the importance of running complete BOS assessments in the same fluid or tissue. Further, low concentrations of BOS in tissues of TB-affected individuals were observed, and significant differences between healthy and sick boar were only detected in the serum of individuals developing mild TB and in the muscle of individuals with mild or severe disease status. However, all organs from wild boars affected with mild TB were not in oxidative imbalance compared to healthy control animals, suggesting that wild boars may cope well with TB. Our data indicate that serum and other tissues can be used as BOS in field conservation programs to monitor wildlife population health. Still, context-specific validations are needed to determine the most appropriate samples to use.

Assessing mammal trapping standards in wild boar drop-net capture.

Applying contemporary trapping standards when managing wildlife should no longer be an option, but a duty. Increasing wild boar populations originate a growing number of conflicts and hunting is the only cost-effective management option in most cases. However, new scenarios where hunting is unfeasible emerge and trapping necessities cope with lacking regulatory frameworks and technical guidelines. In this research, we evaluated drop nets, a capture method not considered by the international trapping standards, to capture Eurasian wild boar (Sus scrofa), a wildlife species not included in the list of mammal species under the scope of the Agreement on International Humane Trapping Standards (AIHTS). Less than 20% of the captured wild boars presented moderate or severe injuries attributable to the capture method, hence fulfilling the acceptance thresholds of the outdated AIHTS. Based on the new standards thresholds of acceptance, the humaneness of drop-nets in our study ranged 66-78%, under the 85% required. The capture success and selectivity were 100%, as ensured by operator-driven triggering, which should be considered the main strengths of this method, together with the minimization of animal suffering owing the short duration of the stressful situation. Additionally, in spite of the socially adverse environment, with people contrary to wild boar removal, no disturbances against the capture system or operations occurred. This is the first assessment of a drop-net capture method according to internationally accepted mammal trapping standards, with unconclusive results. However, there is a need for adapted procedures and thresholds of acceptance aimed at not-mechanical traps in general, and specifically at drop-nets. Compared to other live-capture methods, drop-nets minimize the duration of the stressful situation -at the expense of a strong adrenergic acute response-, maximize the probabilities of capturing entire sounders of prosocial species, which may be also considered as more humane, and has the ability to coordinate higher values of capture success, absolute selectivity and adaptability to difficult environments.

Ivermectin Plasma Concentration in Iberian Ibex (Capra pyrenaica) Following Oral Administration: A Pilot Study.

Sarcoptic mange is considered the main driver of demographic declines occurred in the last decades in Iberian ibex (Capra pyrenaica) populations. Mass treatment campaigns by administration of in-feed acaricides are used as a measure to mitigate the impact of mange in the affected populations. However, there are no data on ivermectin (IVM) pharmacokinetics in this wild caprine, and the treatment through medicated feed is not endorsed by evidence on its effectiveness. The aim of this study is to determine the pharmacokinetic profile of IVM in plasma samples of ibexes after the experimental oral administration of IVM, using high performance liquid chromatography (HPLC) with automated solid phase extraction and fluorescence detection. A dose of 500 µg of IVM per body weight was orally administered in a feed bolus to nine healthy adult ibexes (seven males and two females). Blood samples were collected by jugular venipuncture into heparin-coated tubes at day 1, 2, 3, 4, 7, 10, 15, and 45 post-administration (dpa). The highest plasma concentration of IVM (Cmax = 3.4 ng/ml) was detected 24 h after the oral administration (T1), followed by a rapid decrease during the first week post-administration. Our results reveal that plasma IVM concentration drops drastically within 5 days of ingestion, questioning the effectiveness of a single in-feed dose of this drug to control sarcoptic mange. To the best of our knowledge, this is the first study on plasma availability of oral IVM in ibexes and in any wild ungulate species.

Genetic population structure defines wild boar as an urban exploiter species in Barcelona, Spain.

Urban wildlife ecology is gaining relevance as metropolitan areas grow throughout the world, reducing natural habitats and creating new ecological niches. However, knowledge is still scarce about the colonisation processes of such urban niches, the establishment of new communities, populations and/or species, and the related changes in behaviour and life histories of urban wildlife. Wild boar (Sus scrofa) has successfully colonised urban niches throughout Europe. The aim of this study is to unveil the processes driving the establishment and maintenance of an urban wild boar population by analysing its genetic structure. A set of 19 microsatellite loci was used to test whether urban wild boars in Barcelona, Spain, are an isolated population or if gene flow prevents genetic differentiation between rural and urban wild boars. This knowledge will contribute to the understanding of the effects of synurbisation and the associated management measures on the genetic change of large mammals in urban ecosystems. Despite the unidirectional gene flow from rural to urban areas, the urban wild boars in Barcelona form an island population genotypically differentiated from the surrounding rural ones. The comparison with previous genetic studies of urban wild boar populations suggests that forest patches act as suitable islands for wild boar genetic differentiation. Previous results and the genetic structure of the urban wild boar population in Barcelona classify wild boar as an urban exploiter species. These wild boar peri-urban island populations are responsible for conflict with humans and thus should be managed by reducing the attractiveness of urban areas. The management of peri-urban wild boar populations should aim at reducing migration into urban areas and preventing phenotypic changes (either genetic or plastic) causing habituation of wild boars to humans and urban environments.

Zoonotic Campylobacter spp. and Salmonella spp. carried by wild boars in a metropolitan area: occurrence, antimicrobial susceptibility and public health relevance.

Campylobacter spp. and Salmonella spp. are the most reported zoonotic agents in Europe. They can be transmitted from wildlife to humans, and wild boars (Sus scrofa) can harbour them. In the Metropolitan Area of Barcelona (MAB, NE Spain) wild boars are found in urbanized areas. To assess the potential public health risk of this increasing wild boar population, we collected stool samples from 130 wild boars from the MAB (June 2015 - February 2016), to determine the Campylobacter and Salmonella occurrence and the antimicrobial susceptibility of the isolates. We also investigated the genetic diversity and virulence potential of Campylobacter. Campylobacter prevalence in wild boars was 61%. Forty six percent of wild boars carried Campylobacter lanienae, 16% carried Campylobacter coli, and 1% carried Campylobacter hyointestinalis; 4% carried both C. lanienae and C. coli, and 1% carried both C. lanienae and C. hyointestinalis. This is the first report of C. hyointestinalis in wildlife in Spain. Using pulse-field gel electrophoresis and multilocus sequence typing, we observed a high genetic diversity of Campylobacter and identified new sequence types. Thirty-three percent of C. coli and 14% of C. lanienae isolates showed a high virulence potential. All of the Campylobacter isolates analysed were resistant to at least one antimicrobial agent. Multidrug resistance was only detected in C. coli (67%). Salmonella enterica subsp. enterica was detected in four wild boars (3%) and included a S. Enteritidis serovar (1/4 wild boars) and a multidrug-resistant (ASSuT) monophasic S. Typhimurium serovar (1/4 wild boars) which is associated with human infections and pig meat in Europe. The characteristics of some of the Campylobacter and Salmonella isolates recovered suggest an anthropogenic origin. Wild boars are a reservoir of Campylobacter and have the potential to spread antimicrobial resistant Campylobacter and Salmonella in urbanized areas in the MAB.

Ticks on wild boar in the metropolitan area of Barcelona (Spain) are infected with spotted fever group rickettsiae.

Tick-borne pathogens (TBPs) constitute an emerging public health concern favoured by multidimensional global changes. Amongst these, increase and spread of wild boar (Sus scrofa) populations are of special concern since this species can act as a reservoir of zoonotic pathogens and promote tick abundance. Thus, we aimed to make a first assessment of the risk by TBPs resulting from wild boar and ticks in the vicinity of a highly populated area. Between 2014 and 2016, we collected spleen samples and 2256 ticks from 261 wild boars (out of 438 inspected) in the metropolitan area of Barcelona (MAB; northeast Spain). We morphologically identified four tick species: Hyalomma lusitanicum (infestation prevalence: 33.6%), Dermacentor marginatus (26.9%), Rhipicephalus sanguineus sensu lato (18.9%) and R. bursa (0.2%). Ticks were pooled according to species and individual host. A total of 180 tick pools and 167 spleen samples were screened by real-time PCR and/or reverse line blot hybridization assay for Ehrlichia sp., Anaplasma sp., Babesia sp., Rickettsia sp., Borrelia burgdorferi sensu lato and Coxiella burnetii. Seventy-two out of the 180 tick pools were positive to Rickettsia spp. (minimum prevalence of 8.7%), including Rickettsia massiliae, R. slovaca and R. raoultii. We did not detect Rickettsia spp. in wild boar spleens nor other TBPs in ticks or wild boars. Since the ticks identified can bite humans, and the recorded spotted fever group (SFG) rickettsiae are zoonotic pathogens, there is a risk of SFG rickettsiae transmission for MAB inhabitants. Our results suggest a broader distribution of H. lusitanicum, competent vector for the Crimean-Congo haemorrhagic fever virus than previously known. Wild boar is not a Rickettsia spp. reservoir according to the spleen negative results. However, its abundance could favour tick life cycle and abundance, and its proximity to humans could promote the infection risk by Rickettsia spp.

Assessing the levels of intraspecific admixture and interspecific hybridization in Iberian wild goats (Capra pyrenaica).

Iberian wild goats (Capra pyrenaica, also known as Iberian ibex, Spanish ibex, and Spanish wild goat) underwent strong genetic bottlenecks during the 19th and 20th centuries due to overhunting and habitat destruction. From the 1970s to 1990s, augmentation translocations were frequently carried out to restock Iberian wild goat populations (very often with hunting purposes), but they were not systematically planned or recorded. On the other hand, recent data suggest the occurrence of hybridization events between Iberian wild goats and domestic goats (Capra hircus). Augmentation translocations and interspecific hybridization might have contributed to increase the diversity of Iberian wild goats. With the aim of investigating this issue, we have genotyped 118 Iberian wild goats from Tortosa-Beceite, Sierra Nevada, Muela de Cortes, Gredos, Batuecas, and Ordesa and Monte Perdido by using the Goat SNP50 BeadChip (Illumina). The analysis of genotypic data indicated that Iberian wild goat populations are strongly differentiated and display low diversity. Only three Iberian wild goats out from 118 show genomic signatures of mixed ancestry, a result consistent with a scenario in which past augmentation translocations have had a limited impact on the diversity of Iberian wild goats. Besides, we have detected eight Iberian wild goats from Tortosa-Beceite with signs of domestic goat introgression. Although rare, hybridization with domestic goats could become a potential threat to the genetic integrity of Iberian wild goats; hence, measures should be taken to avoid the presence of uncontrolled herds of domestic or feral goats in mountainous areas inhabited by this iconic wild ungulate.

First report of interspecific transmission of sarcoptic mange from Iberian ibex to wild boar.

BACKGROUND: Sarcoptic mange is a globally distributed parasitic disease caused by the burrowing mite Sarcoptes scabiei. This mite has a certain degree of host specificity, although interspecific transmission can occur among phylogenetically related species or through prey-predator mediated exposure. In 2018, a wild boar (Sus scrofa) with lesions compatible with sarcoptic mange was hunted in Ports de Tortosa i Beseit Natural Park (PTB, north-eastern Spain), where an active epizootic outbreak of sarcoptic mange is affecting Iberian ibexes (Capra pyrenaica) since 2014. METHODS: A complete necropsy, skin scrapings and skin digestions with hydroxide potassium were performed to confirm the diagnosis. Routine histopathological analysis, toluidine blue staining and immunohistochemistry were used to characterize the lesions and the inflammatory infiltrate. Finally, 10 specific S. scabiei microsatellites were molecularly genotyped through polymerase chain reactions in mites obtained from the affected wild boar. For phylogenetic comparison, mites obtained from sympatric Iberian ibexes and allopatric wild boars and Iberian ibexes from southern Spain were analysed. RESULTS: Sarcoptes scabiei was visually and molecularly identified in the infested wild boar from PTB, causing skin lesions with dermal inflammatory infiltrate rich in T and B cells, which indicate an adaptive immune response. Three S. scabiei genetic clusters were identified: one included mites from southern Iberian ibexes, another included mites from southern wild boars, and a third one distinctively grouped the wild boar from PTB with the sympatric ibexes. CONCLUSIONS: To the authors' knowledge, this is the first reported case of sarcoptic mange in wild boar in Spain and the first documented case of S. scabiei cross-transmission from a wild ruminant host to a wild boar. The wild boar presented an ordinary scabies type reaction, which is typical of the self-limiting infestations reported in other cases of interspecific transmission.

Detection of Beta-Lactam-Resistant Escherichia coli and Toxigenic Clostridioides difficile Strains in Wild Boars Foraging in an Anthropization Gradient.

Disease transmission among wild boars, domestic animals and humans is a public health concern, especially in areas with high wild boar densities. In this study, fecal samples of wild boars (n = 200) from different locations of the Metropolitan Area of Barcelona were analyzed by PCR to explore the frequency of ß-lactamases and extended cephalosporin and carbapenem resistance genes (ESBLs) in Escherichia coli strains and the presence of toxigenic Clostridioides difficile. The prevalence of genes conferring resistance to ß-lactam antimicrobials was 8.0% (16/200): blaCMY-2 (3.0%), blaTEM-1b (2.5%), blaCTX-M-14 (1.0%), blaSHV-28 (1.0%), blaCTX-M-15 (0.5%) and blaCMY-1 (0.5%). Clostridioides difficile TcdA+ was detected in two wild boars (1.0%), which is the first report of this pathogen in wild boars in Spain. Moreover, the wild boars foraging in urban and peri-urban locations were more exposed to AMRB sources than the wild boars dwelling in natural environments. In conclusion, the detection of E. coli carrying ESBL/AmpC genes and toxigenic C. difficile in wild boars foraging in urban areas reinforces the value of this game species as a sentinel of environmental AMRB sources. In addition, these wild boars can be a public and environmental health concern by disseminating AMRB and other zoonotic agents. Although this study provides the first hints of the potential anthropogenic sources of AMR, further efforts should be conducted to identify and control them.

Wild boar in the city: Phenotypic responses to urbanisation.

Urbanisation is a global human-induced environmental change and one of the most important threats to biodiversity. To survive in human-modified environments, wildlife must adjust to the challenging selection pressures of urban areas through behaviour, morphology, physiology and/or genetic changes. Here we explore the effect of urbanisation in a large, highly adaptable and generalist urban adapter species, the wild boar (Sus scrofa, Linnaeus 1758). From 2005 to 2018, we gathered wild boar data and samples from three areas in NE Spain: one urban (Barcelona municipality, n = 445), and two non-urban (Serra de Collserola Natural Park, n = 183, and Sant Llorenç del Munt i Serra de l'Obac Natural Park, n = 54). We investigated whether urbanisation influenced wild boar body size, body mass, body condition, and the concentration of serum metabolites, considering also the effect of age, sex and use of anthropogenic food resources. Wild boars from the urban area had larger body size, higher body mass, better body condition, and a higher triglyceride and lower creatinine serum concentrations than non-urban wild boars. In addition, urban wild boars consumed food from anthropogenic origin more frequently, which suggests that differences in their diet probably induced the biometric and the metabolic changes observed. These responses are probably adaptive and suggest that wild boars are thriving in the urban environment. Our results show that urbanisation can change the morphological and physiological traits of a large mammal urban adapter, which may have consequences in the ecology and response to urban selection pressures by the species. The phenotypic plasticity shown by wild boars provides both further and new evidence on the mechanisms that allow urban adapter species of greater size to respond to urbanisation, which is expected to continue growing globally over the coming decades.

Molecular Detection and Identification of Chlamydiaceae in the Eyes of Wild and Domestic Ruminant Hosts from Northern Spain.

Infections by Chlamydiae are associated with ocular disease in humans and animals. In this study, the presence and diversity of Chlamydia spp. was assessed in diseased and healthy eyes of domestic sheep and wild ruminants that share mountain habitats in northern Spain. The presence of Chlamydia spp. was tested by real-time PCR in 1786 conjunctival swabs collected from both eyes of 893 animals from mountain habitats in northern Spain, and chlamydial species were identified in the positive samples by ArrayTube microarray methods. Chlamydial DNA was detected in 0.6% (CI95% 0.2-1.3) of the Pyrenean chamois (Rupicapra pyrenaica) and 1.4% (CI95% <0.01-8.1) of the sheep (Ovis aries) sampled, with Chlamydia pecorum the only chlamydial species identified. No association of C. pecorum with ocular disease or co-infection with Mycoplasma conjunctivae was found. Further studies on the pathogenesis of infectious keratoconjunctivitis are needed to better understand the ecology of C. pecorum and its possible role as a ruminant pathogen at the wildlife-livestock interface.

Sarcoptic mange in wild ruminants in Spain: solving the epidemiological enigma using microsatellite markers.

BACKGROUND: In Spain, sarcoptic mange was first described in native wildlife in 1987 in Cazorla Natural Park, causing the death of nearly 95% of the local native population of Iberian ibex (Capra pyrenaica). Since then, additional outbreaks have been identified in several populations of ibex and other wild ungulate species throughout the country. Although the first epizootic outbreak in wildlife was attributed to the introduction of an infected herd of domestic goats, the origin and the cause of its persistence remain unclear. The main aims of this study are to understand (i) the number of Sarcoptes scabiei "strains" circulating in wild ruminant populations in Spain, and (ii) the molecular epidemiological relationships between S. scabiei and its hosts. METHODS: Ten Sarcoptes microsatellite markers were used to characterize the genetic structure of 266 mites obtained from skin scrapings of 121 mangy wild ruminants between 2011 and 2019 from 11 areas in Spain. RESULTS: Seventy-three different alleles and 37 private alleles were detected. The results of this study show the existence of three genetic strains of S. scabiei in the wild ruminant populations investigated. While two genetic clusters of S. scabiei were host- and geography-related, one cluster included multi-host mites deriving from geographically distant populations. CONCLUSIONS: The molecular epidemiological study of S. scabiei in wild ruminants in Spain indicates that the spreading and persistence of the parasite may be conditioned by host species community composition and the permissiveness of each host population/community to the circulation of individual "strains," among other factors. Wildlife-livestock interactions and the role of human-driven introduction or trade of wild and domestic animals should be better investigated to prevent further spread of sarcoptic mange in as yet unaffected natural areas of the Iberian Peninsula.

Endemic occurrence of Fasciola hepatica in an alpine ecosystem, Pyrenees, Northeastern Spain.

Fasciola hepatica is a liver parasite of ruminants whose distribution is determined by its intermediate host, the freshwater snail Galba truncatula. In Europe, F. hepatica is mostly associated with lowlands. Infection from sympatric domestic reservoirs is rarely reported in wild mountain ungulates. This study explores F. hepatica in a multi-host system in a European alpine area. Serum samples (n = 1,209) from Pyrenean chamois (Rupicapra p. pyrenaica), European mouflon (Ovis aries musimon), domestic sheep (Ovis aries) and domestic cattle (Bos taurus) were collected in the National Game Reserve of Freser-Setcases (NGRFS) in Catalonia, Northeastern Spain, from 2008 to 2019, and tested for antibodies against F. hepatica. During the same period, the livers of 214 chamois hunted in the NGRFS were inspected for F. hepatica and associated pathological changes. Finally, 907 freshwater snails were collected in summer 2016 between 1559 and 2,224 metres above sea level (asl) in the NGRFS, and F. hepatica DNA sought by PCR. Antibodies against F. hepatica were detected in all four species, with a higher prevalence in cattle and sheep than in chamois. Fasciola hepatica and hepatic lesions were concurrently observed in 13/214 of the chamois livers inspected (6.1%, CI95 2.9%-9.3%). Fasciola hepatica DNA was detected in one out of the 907 snails (0.1%, Cl95 0.1% - 0.3%; Ct value 33.3) and collected at 2054 m asl. Fasciola hepatica was consistently detected in a high mountain multi-host system, suggesting that its life cycle is completed and that it occurs endemically at the highest elevation reported in Europe. Transhumant livestock are the likely source in this alpine ecosystem, which according to rare occurrence of F. hepatica DNA in G. truncatula is still a suboptimal habitat for F. hepatica life cycle. Studying parasites at their highest distribution range can be useful to monitor climate change in seasonal mountain environments.

Assessing methods to live-capture wild boars (Sus scrofa) in urban and peri-urban environments.

BACKGROUND: Wild boar (Sus scrofa) populations are increasing worldwide and invading urban areas. Live-capture can improve the management of this challenge, maximising efficiency, allowing scientific studies and potentially improving animal welfare. This study assesses teleanaesthesia, drop-net, corral trap and cage trap to live-capture wild boar in urban and peri-urban areas, evaluating efficiency and animal stress through haematology and serum biochemistry. METHODS: From 2012 to 2018, 655 wild boars were captured in 279 operations (drop-net=17, teleanaesthesia=186, cage trap=66 and corral trap=10) in the urban and peri-urban areas of Barcelona (Spain). Haematological and serum biochemical variables were determined in 145 wild boars (42 drop-netted, 41 teleanaesthetised, 38 cage-trapped and 24 corral-trapped). RESULTS: Performance (wild boars captured per operation) was highest for drop-net, followed by corral and cage traps, and finally teleanaesthesia. The three physical capture methods were more stressful than teleanaesthesia, causing a more intense physiological reaction, muscular damage, renal function impairment and homeostasis adaption. Stress response was predominantly adrenergic for drop-net and cortisol-induced for cage and corral traps. CONCLUSION: Teleanaesthesia is the choice in reactive urban situations thanks to its adaptability; drop-net effectively targets wild boars in peri-urban environments; cage and corral traps are useful as long-term methods in specific areas.

Accumulation of diastereomers of anticoagulant rodenticides in wild boar from suburban areas: Implications for human consumers.

We studied the prevalence of anticoagulant rodenticides (ARs) in liver and muscle tissues of wild boar captured in the urban area of Barcelona, the suburban area of Collserola Natural Park and the rural area of Santa Quiteria, next to Cabañeros National Park, in Spain. The objective was to assess the influence of both urbanisation and wild boar (Sus scrofa) trophic opportunism on the accumulation of these compounds. We have also evaluated the risk for human consumers of this game meat. Wild boars from Barcelona city showed the highest prevalence of ARs detection (60.8%), followed by the adjoining suburban area of Collserola N.P. (40%) and the rural distant area of Santa Quiteria (7.7%). Liver bioaccumulated ARs (45.2%) more frequently than muscle (11.9%). A significant proportion (13.7%) of wild boar captured in Barcelona city exceeded 200 ng/g of total ARs in liver, a threshold for adverse effects on blood clotting. For difenacoum, there was a predominance of cis isomer, while for brodifacoum and bromadiolone cis and trans isomers appeared in a similar proportion. According to the scarce available information on ARs toxicity in humans, the risk of acute poisoning from game meat consumption seems to be low. However, repeated exposure through liver consumption should be considered in further risk assessments because of the high concentration detected in some samples (up to 0.68 mg/kg).