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 The systematic review of all the information on diseases affecting the species unveils unpublished reports, scientific communications in meetings, and scientific articles, allowing the first comprehensive compilation of Iberian ibex diseases.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.

Generalized tuberculosis due to Mycobacterium caprae in a red fox phylogenetically related to livestock breakdowns.

BACKGROUND: Tuberculosis (TB) due to Mycobacterium caprae is endemic in goat herds and free-ranging wild boars in Spain, causing infections in other livestock or wild animals to a lesser extent. TB infection in foxes is infrequently reported and they are usually considered spillover hosts of TB. CASE PRESENTATION: A blind, depressed and severely emaciated red fox (Vulpes vulpes) was admitted to a rehabilitation center. After clinical examination it was humanely sacrificed. At necropsy, generalized TB lesions were observed that were subsequently confirmed by histopathology along with a co-infection with canine distemper virus. M. caprae was isolated from mycobacterial culture and spoligotype SB0415 was identified. Whole genome sequencing (WGS) of the isolated M. caprae was carried out and single nucleotide polymorphisms (SNP) were compared with other sequences of M. caprae isolated from livestock and wildlife of the same area throughout the last decade. CONCLUSIONS: This is the first reported case of TB due to M. caprae in a fox in the Iberian Peninsula. WGS and SNP analysis, together with spatial-temporal investigations, associated this case with recent M. caprae outbreaks in cattle and goat herds of the area. The results indicated transmission of M. caprae between livestock and the fox, suggesting that this species may occasionally play a role in the epidemiology of animal TB.

Urban Wild Boars and Risk for Zoonotic Streptococcus suis, Spain.

Urban wild boars (Sus scrofa) from Barcelona, Spain, harbor great diversity of Streptococcus suis strains, including strains with the cps2 gene and with the same molecular profile as local human cases. The increasing trend of potential effective contacts for S. suis transmission is of public health concern.

Postepizootic Persistence of Asymptomatic Mycoplasma conjunctivae Infection in Iberian Ibex.

The susceptibility of the Iberian ibex (Capra pyrenaica) to Mycoplasma conjunctivae ocular infection and the changes in their interaction over time were studied in terms of clinical outcome, molecular detection, and IgG immune response in a captive population that underwent a severe infectious keratoconjunctivitis (IKC) outbreak. Mycoplasma conjunctivae was detected in the Iberian ibex, coinciding with the IKC outbreak. Its prevalence had a decreasing trend in 2013 that was consistent with the clinical resolution (August, 35.4%; September, 8.7%; November, 4.3%). Infections without clinical outcome were, however, still detected in the last handling in November. Sequencing and cluster analyses of the M. conjunctivae strains found 1 year later in the ibex population confirmed the persistence of the same strain lineage that caused the IKC outbreak but with a high prevalence (75.3%) of mostly asymptomatic infections and with lower DNA load of M. conjunctivae in the eyes (mean quantitative PCR [qPCR] cycle threshold [CT ], 36.1 versus 20.3 in severe IKC). Significant age-related differences of M. conjunctivae prevalence were observed only under IKC epizootic conditions. No substantial effect of systemic IgG on M. conjunctivae DNA in the eye was evidenced with a linear mixed-models selection, which indicated that systemic IgG does not necessarily drive the resolution of M. conjunctivae infection and does not explain the epidemiological changes observed. The results show how both epidemiological scenarios, i.e., severe IKC outbreak and mostly asymptomatic infections, can consecutively occur by entailing mycoplasma persistence.IMPORTANCEMycoplasma infections are reported in a wide range of epidemiological scenarios that involve severe disease to asymptomatic infections. This study allows a better understanding of the transition between two different Mycoplasma conjunctivae epidemiological scenarios described in wild host populations and highlights the ability of M. conjunctivae to adapt, persist, and establish diverse interactions with its hosts. The proportion of asymptomatic and clinical M. conjunctivae infections in a host population may not be regarded only in response to intrinsic host species traits (i.e., susceptibility) but also to a specific host-pathogen interaction, which in turn influences the infection dynamics. Both epidemic infectious keratoconjunctivitis and a high prevalence of asymptomatic M. conjunctivae infections may occur in the same host population, depending on the circulation of M. conjunctivae, its maintenance, and the progression of the host-pathogen interactions.

Identification of a gammaherpesvirus belonging to the malignant catarrhal fever group of viruses in Pyrenean chamois (Rupicapra p. pyrenaica).

High prevalence (46 %) of a gammaherpesvirus was confirmed by molecular detection in the lungs of hunted Pyrenean chamois. The partial glycoprotein B sequence up to the DNA polymerase gene showed 96.6 % nucleotide sequence identity to the Rupicapra rupicapra gammaherpesvirus 1 and 81.5 % to ovine herpesvirus 2. This novel sequence clusters within sequences derived from the malignant catarrhal fever group of viruses, and the corresponding virus is tentatively named Rupicapra pyrenaica gammaherpesvirus 1 (RpHV-1). No specific histological lesions were associated with RpHV-1, nor were any detrimental effects on host health. The epidemiological, phylogenetic and histopathological results suggest that Pyrenean chamois is the natural host of RpHV-1.

Coprological tests underestimate Macracanthorhynchus hirudinaceus burden in wild boar.

The present study evaluated the limitations of the coprological sedimentation test to assess Macracanthorhynchus hirudinaceus infestation in 59 wild boars (Sus scrofa) from central Spain. The coprological sedimentation test appeared to be a poor predictor of both prevalence of infestation and the real parasite burden due to the high number of false negative results (prevalence was reduced from 61 to 16 %). Because of the potential increased risk of this zoonosis, it is suggested that alternative techniques be used in wildlife surveillance programmes.

Lead Poisoning Due to Lead-Pellet Ingestion in Griffon Vultures ( Gyps fulvus ) From the Iberian Peninsula.

Avian scavengers that typically include game birds and mammals in their diets are at risk of lead poisoning from ingestion of carcasses with fragmented or residual lead ammunition that is used in hunting. Thus, lead may be one of the threats that the griffon vulture ( Gyps fulvus ) faces in the Iberian Peninsula and particularly in Portugal, where their conservation status is considered to be near-threatened. This is the first report that details 3 cases of lead poisoning, associated with the ingestion of lead shot, in adult female griffon vultures found in the Iberian Peninsula. The birds were found prostrate and immediately transferred to a wildlife rehabilitation center, where they died within 24 hours after supportive treatment. Necropsy and histopathologic examinations were done in 2 birds and metal analyses were done in all birds to determine the birds' causes of death. In one vulture, 9 uneroded lead pellets were recovered from the stomach, and moderate to severe hemosiderosis was seen histologically in the liver, lungs, and kidneys. Diagnosis of lead poisoning was confirmed by results of metal analyses, which revealed extremely high lead concentrations in blood (969-1384 µg/dL), liver (309-1077 µg/g dry weight), and kidneys (36-100 µg/g dry weight) for all 3 vultures. To prevent lead poisoning in vultures and preserve their populations in the Iberian Peninsula, more resources are needed for diagnosis and treatment of wildlife in rehabilitation centers, new regulations enabling the abandonment of fallen stock in the field must be approved, and lead ammunition must be prohibited in big-game hunting.

Factors associated with the prevalence and pathology of Calodium hepaticum and C. splenaecum in periurban micromammals.

Calodium hepaticum (syn. Capillaria hepatica) and Calodium splenaecum (syn. Capillaria splenaecum) are nematodes that infect the liver and spleen, respectively, of mammals. While the host range, distribution, pathology and zoonotic potential of C. hepaticum are well known, very little is known about C. splenaecum. The observed prevalence of these two parasites, the factors associated with prevalence, and the lesions resulting in the different host species were studied in 408 micromammals captured in two periurban areas of Barcelona (NE Spain) from 2011 to 2013. C. hepaticum was found in 4% of 322 wood mice (Apodemus sylvaticus) (with local prevalence up to 16%) and 1 of 2 Norwegian rats (Rattus norvegicus). C. splenaecum was found in 10 % of 38 greater white-toothed shrew (Crocidura russula) (local prevalence up to 30%). Neither parasite was detected in 29 Algerian mice (Mus spretus) and 17 black rats (Rattus rattus). Prevalence of C. hepaticum was significantly higher in wood mice captured in natural areas (6.4%) than those from residential areas (0%), and infected mice were in better body condition. No differences in prevalence were found among age and sex groups, years and seasons. Lesions of hepatic capillariasis in wood mice consisted mainly of mild to moderate multifocal granulomas around degenerating adult parasites and/or eggs, while lesions seen in a rat consisted of multifocal granulomatous hepatitis and bridging fibrosis extending from the necrotic areas caused by the parasites. Splenic lesions found in shrews due to C. splenaecum, representing the first histological description of this parasite, were single nodules that corresponded to finely encapsulated clusters of eggs with adult parasites.

An identification key for the five most common species of Metastrongylus.

Species of the Metastrongylus genus, the lung nematodes of pigs that require an intermediate host (earthworm) to complete their cycle, pose a potential risk to both livestock and humans. This parasite which can result in lung pathology and mixed infections with other pathogens (e.g. viruses) can be fatal to pigs. Although this genus is distributed worldwide, there are no classification keys for identifying this common parasite species. In this work, we take advantage of parasitological surveys of wild boar (Sus scrofa) in northern and central Spain and southern Poland to develop a morphological identification key for the five most common Metastrongylus species (Metastrongylus apri, Metastrongylus pudendotectus, Metastrongylus salmi, Metastrongylus confusus and Metastrongylus asymetricus). In addition, we provide the first record of M. confusus in Spain, probably unidentified until now due to the lack of appropriate identification keys. We hope that this user-friendly identification key will enable parasitologists and veterinary practitioners to avoid further misclassifications of Metastrongylus species.

Recombination between non-structural and structural genes as a mechanism of selection in lagoviruses: The evolutionary dead-end of an RHDV2 isolated from European hare.

The genus Lagovirus, belonging to the family Caliciviridae, emerged around the 1980s. It includes highly pathogenic species, rabbit hemorrhagic disease virus (RHDV/GI.1) and European brown hare syndrome virus (EBHSV/GII.1), which cause fatal hepatitis, and nonpathogenic viruses with enteric tropism, rabbit calicivirus (RCV/GI.3,4) and hare calicivirus (HaCV/GII.2). Lagoviruses have evolved along two independent genetic lineages: GI (RHDV and RCV) in rabbits and GII (EBHSV and HaCV) in hares. To be emphasized is that genomes of lagoviruses, like other caliciviruses, are highly conserved at RdRp-VP60 junctions, favoring intergenotypic recombination events at this point. The recombination between an RCV (genotype GI.3), donor of non-structural (NS) genes, and an unknown virus, donor of structural (S) genes, likely led to the emergence of a new lagovirus in the European rabbit, called RHDV type 2 (GI.2), identified in Europe in 2010. New RHDV2 intergenotypic recombinants isolated in rabbits in Europe and Australia originated from similar events between RHDV2 (GI.2) and RHDV (GI.1) or RCV (GI.3,4). RHDV2 (GI.2) rapidly spread worldwide, replacing RHDV and showing several lagomorph species as secondary hosts. The recombination events in RHDV2 viruses have led to a number of viruses with very different combinations of NS and S genes. Recombinant RHDV2 with NS genes from hare lineage (GII) was recently identified in the European hare. This study investigated the first RHDV2 (GI.2) identified in Italy in European hare (RHDV2_Bg12), demonstrating that it was a new virus that originated from the recombination between RHDV2, as an S-gene donor and a hare lagovirus, not yet identified but presumably nonpathogenic, as an NS gene donor. When rabbits were inoculated with RHDV2_Bg12, neither deaths nor seroconversions were recorded, demonstrating that RHDV2_Bg12 cannot infect the rabbit. Furthermore, despite intensive and continuous field surveillance, RHDV2_Bg12 has never again been identified in either hares or rabbits in Italy or elsewhere. This result showed that the host specificity of lagoviruses can depend not only on S genes, as expected until today, but potentially also on some species-specific NS gene sequences. Therefore, because RHDV2 (GI.2) infects several lagomorphs, which in turn probably harbor several specific nonpathogenic lagoviruses, the possibility of new speciation, especially in those other than rabbits, is real. RHDV2 Bg_12 demonstrated this, although the attempt apparently failed.

Adipocytes in synovial fluid cytology: An approach for diagnosing synovial lipomatosis.

A 2-year-old neutered male bullmastiff dog was presented with chronic left hind limb lameness. Physical examination revealed left stifle effusion and medial buttress without cranial tibial thrust. Radiographs showed joint effusion and new bone formation at the patella apex. Magnetic resonance imaging showed increased synovial fluid, widening of the joint space, abnormal infrapatellar fat body and thinning of the cranial cruciate ligament. Synoviocentesis and cytologic evaluation of synovial fluid revealed marked mononuclear inflammation with abundant fatty tissue, suggesting synovial lipomatosis in conjunction with the imaging findings. The disease was confirmed histologically after sampling the lesion during arthrotomy. Synovial lipomatosis, characterized by extensive synovial adipose tissue proliferation of the synovial membrane, is a rare "tumor-like" disorder that usually affects the stifle. Although the etiology remains unclear, joint trauma, inflammation, instability, and lipid abnormalities have been proposed as causes. Inflammatory factors may promote synoviocyte and adipocyte hyperplasia that perpetuate the process. Surgical removal may be suggested to eliminate triggers and prevent future recurrences. The report provides the first cytological description of adipocytes in synovial fluid associated with the diagnosis of synovial lipomatosis in dogs. This case report underscores the potential effectiveness of cytologic analysis of synovial fluid smears, in combination with magnetic resonance imaging (MRI), for diagnosing this condition and reducing complications associated with arthrotomy for sampling purposes. Additionally, the case highlights that synovial lipomatosis should be considered as a potential differential diagnosis for synovial masses in dogs. Further cases are needed to validate these observations in veterinary medicine.

First report of Sarcocystis halieti (Apicomplexa) in bearded vulture (Gypaetus barbatus).

At least three Sarcocystis species (S. falcatula, S. halieti and S. wobeseri-like) have been detected infecting raptorial birds. By histopathology and PCR-sequencing of the ITS1 marker, S. halieti was detected in a bearded vulture (Gypaetus barbatus) and a black kite (Milvus migrans) from the Catalonia region in North Spain. The 241 bp-long sequences obtained from the Sarcocystis organisms detected in both raptors showed 97.5-99.6% and 97.9-100% similarity with those of previously identified S. halieti; also, the phylogenetic trees generated placed the identified sequences together with other sequences of S. halieti available in GenBank. In sum, the description of the bearded vulture as a new intermediate host for S. halieti adds new insights on the complex epidemiology of the genus involving avian hosts.

A new HaCV-EBHSV recombinant lagovirus circulating in European brown hares (Lepus europaeus) from Catalonia, Spain.

In 2020/2021, several European brown hare syndrome virus (EBHSV) outbreaks were recorded in European hares (Lepus europaeus) from Catalonia, Spain. Recombination analysis combined with phylogenetic reconstruction and estimation of genetic distances of the complete coding sequences revealed that 5 strains were recombinants. The recombination breakpoint is located within the non-structural protein 2C-like RNA helicase (nucleotide position ~ 1889). For the genomic fragment upstream of the breakpoint, a non-pathogenic EBHSV-related strain (hare calicivirus, HaCV; GII.2) was the most closely related sequence; for the rest of the genome, the most similar strains were the European brown hare syndrome virus (EBHSV) strains recovered from the same 2020/2021 outbreaks, suggesting a recent origin. While the functional impact of the atypical recombination breakpoint remains undetermined, the novel recombinant strain was detected in different European brown hare populations from Catalonia, located 20-100 km apart, and seems to have caused a fatal disease both in juvenile and adult animals, confirming its viability and ability to spread and establish infection. This is the first report of a recombination event involving HaCV and EBHSV and, despite the recombination with a non-pathogenic strain, it appears to be associated with mortality in European brown hares, which warrants close monitoring.

Leishmania Infection in Wild Lagomorphs and Domestic Dogs in North-East Spain.

Leishmania infantum is a zoonotic protozoan parasite distributed worldwide that is transmitted by phlebotomine sandflies. Dogs are the main reservoir for human infections. However, in recent years, the capacity of lagomorphs to contribute to Leishmania transmission has been confirmed. The present study aimed to assess Leishmania spp. exposure and infection in lagomorphs and sympatric domestic dogs in NE Spain. Sera from European hares, European rabbits, and rural dogs were tested for antibodies against L. infantum using an in-house indirect ELISA. PCR analysis targeting Leishmania spp. was performed in spleens from L. europaeus. Antibodies against Leishmania spp. were detected in all the species analyzed. Total sample prevalence was significantly higher in O. cuniculus (27.9%) than in L. europaeus (2.0%). Results of the PCR were all negative. The present study expands knowledge about Leishmania infections in free-ranging lagomorphs in the Iberian Peninsula, suggesting a more important role of O. cuniculus in the study area. Given the strong correlation between lagomorph densities and human leishmaniasis outbreaks in Spain, the high rabbit and human densities in NE Spain, and the high Leishmania spp. seroprevalence in rabbits, it becomes imperative to establish surveillance programs for lagomorphs in this region.

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.

First detection of herpesvirus and hemosporidians in the endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus).

Pathogens affect wild bird populations worldwide, contributing to their decline. Considering the scarce health data regarding the endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus), we molecularly surveyed selected pathogens (Newcastle disease virus, Avian influenza virus, Chlamydia psittaci, avian pathogenic Escherichia coli, Campylobacter jejuni, and Salmonella spp.) in 30 Pyrenean Capercaillie feces collected in the field (Catalonia, northeastern Spain). Additionally, swab and tissue samples from eight wild Pyrenean Capercaillies of Catalonia and Andorra were molecularly tested for herpesvirus and hemosporidians (Plasmodium spp., Haemoproteus spp., and Leucocytozoon spp.). All fecal samples were negative for the pathogens tested. Nevertheless, we detected a novel herpesvirus in 50% (4/8) of the Pyrenean Capercaillies, and hemosporidian DNA in 62.5% (5/8) of the tissue samples (i.e., Haemoproteus sp. [4 of 8] and/or Leucocytozoon sp. [3 of 8]). To our knowledge, this is the first detection of herpesvirus and hemosporidians infections in Pyrenean Capercaillies. The putative novel herpesvirus belongs to the genus Iltovirus. The presence of hemosporidian parasites in this mountain bird species is of concern, and could be related to the marked increase in the average temperature in the Pyrenees as a consequence of climate change. Our findings are fundamental to improve the conservation plans for the endangered Pyrenean Capercaillie population.

Coinfection of Chytrid Fungi in Urodeles during an Outbreak of Chytridiomycosis in Spain.

Chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal), has had an unprecedented impact on amphibian biodiversity. Although Bd is globally widespread, Bsal is currently spreading, increasing the probability that these pathogens will co-occur in individual amphibians. Interactions among coinfecting parasites can have significant outcomes on disease dynamics and impact and, therefore, may have important consequences for amphibian conservation. We analyzed the patterns of Bd-Bsal coinfections in two species of free-ranging urodeles during an outbreak of chytridiomycosis in Spain. Our goals were to assess 1) the probability of co-occurrence of both chytrid species and 2) the correlation of pathogen loads in coinfected hosts. We detected coinfections in 81.58% of Triturus marmoratus (n=38) and in 18.75% of Ichthyosaura alpestris (n=16). Histopathologic lesions of chytridiomycosis were observed only in T. marmoratus. Our results demonstrate a positive relationship between Bd and Bsal loads in T. marmoratus, whereas the co-occurrence analysis showed a random association among pathogens in both urodele species. Overall, we show that Bd-Bsal coinfections intensify pathogen load in T. marmoratus and could, therefore, increase disease severity and have important consequences for the conservation of some amphibian species.

Absence of anti-Besnoitia spp. specific antibodies in European wild lagomorphs from the Iberian Peninsula.

Besnoitia besnoiti is an apicomplexan parasite whose life cycle is not completely understood. It is assumed that this parasite might have an indirect life cycle with a carnivore as a definitive host able to shed oocysts after the ingestion of mature cysts in tissues of an infected intermediate host. Cattle and wild cervids on the Iberian Peninsula can act as intermediate hosts of B. besnoiti, and exposure to the parasite has been demonstrated in equids. In this study, we aimed to assess the presence of members of the genera Besnoitia in wild lagomorphs from the Iberian Peninsula and the potential role of these host species in the life cycle of B. besnoiti, as all the animals were sampled from 19 regions of the Iberian Peninsula where cases of bovine besnoitiosis have been previously detected. Serum samples (Oryctolagus cuniculus: n = 552; Lepus europaeus: n = 122) were first analysed by ELISA and subsequently confirmed by Western blot (WB). Specific antibodies against B. besnoiti were not found in any sampled animal by WB. In addition, lung samples from a subset of wild rabbits (n = 16) were tested by PCR and Besnoitia spp. DNA was not detected. These results suggest that Besnoitia spp. are unlikely to circulate in wild lagomorphs in the Iberian Peninsula. Thus, lagomorphs are not expected to play a key role in the biological cycle of B. besnoiti. Further studies are necessary to assess whether different micromammal species, such as rodents, can serve as natural reservoirs of Besnoitia spp. in other European regions.

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.

Experimental infection with chamois border disease virus causes long-lasting viraemia and disease in Pyrenean chamois (Rupicapra pyrenaica).

Since 2001, severe outbreaks of disease associated with border disease virus (BDV) infection have been reported in Pyrenean chamois. The disease is characterized by variable degrees of cachexia, alopecia and neurological manifestations prior to death. The aim of this study was to investigate this disease under experimental conditions. To assess viral virulence, humoral immune response, dissemination and probable routes of transmission, seven chamois (five seronegative and two seropositive for BDV) were inoculated with a BDV isolated from a naturally infected chamois. A group of three chamois were maintained as uninfected controls. The five seronegative chamois became viraemic from day 2 post-inoculation (p.i.) until their death (three animals) or the end of the experiment (on day 34 p.i.) and developed neutralizing antibodies from day 18 p.i. until the end of the study. Continuous shedding of the virus was detected by RT-PCR in oral, nasal and rectal swabs in viraemic chamois from day 5 p.i. Despite none of the viraemic chamois showing obvious neurological signs, all of them had a non-suppurative meningoencephalitis as seen in naturally infected chamois. The two inoculated BDV-seropositive chamois did not become viraemic. This study confirms that BDV is the primary agent of the disease that has been affecting chamois populations in recent years in the Pyrenees and that previously acquired humoral immunity is protective.