UNILATERAL SINGLE VAGINAL ECTOPIC URETER WITH IPSILATERAL HYPOPLASTIC AND DEGENERATED KIDNEY ASSOCIATED WITH INFERTILITY IN IBERIAN IBEX (CAPRA PYRENAICA) DOES.

This article describes the urinogenital condition of three female Iberian ibexes (Capra pyrenaica-one infertile 3-yr-old adult and two prepubertal animals aged 1 (PP1) and 2 (PP2) yr, respectively, all raised in captivity. All showed constant urinal dribbling, leading to ulcerative dermatitis in the vulvar area. Housed in a stable with other females, the adult did not become pregnant after male contact in either of two consecutive mating seasons. Vaginoscopy and laparoscopic exploration performed on the prepubertal females revealed abnormalities of the vagina and urinary bladder. Ultrasound examination revealed atrophy of the left kidney in the adult female and PP1, and of the right kidney in PP2, with degeneration of the renal pelvis. A paraovarian cyst with hydrosalpinx was also detected in the left oviduct of the adult female. Postmortem analysis of the adult and PP2, which shared a mother, confirmed an extramural single ectopic ureter with vaginal insertion associated with atrophy of the ipsilateral kidney. Though PP1 was officially unrelated to the latter animals, all three might have had a common ancestor in their lineages.

Individual stress responses to Sarcoptes scabiei infestation in Iberian ibex, Capra pyrenaica.

In this study we have monitored the stress of Iberian ibex at individual level within the course of an experimental infection with Sarcoptes scabiei mites. For this purpose we have measured faecal 11-ketoetiocholanolone (11-k) using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). We used linear mixed models to explore the effects of host sex and age, clinic (mange status) and time (number of days post-infection) on the concentration of faecal 11-k. The most parsimonious model included clinic, time and host age, which explained 76.6% of the variance of the response variable. Moreover, the concentration of faecal 11-k varied greatly between individuals. Our results evidence the stressor nature of the disease and highlight the negative effects on hosts due to cortisol release and activity.

Hidden MHC genetic diversity in the Iberian ibex (Capra pyrenaica).

BACKGROUND: Defining hidden genetic diversity within species is of great significance when attempting to maintain the evolutionary potential of natural populations and conduct appropriate management. Our hypothesis is that isolated (and eventually small) wild animal populations hide unexpected genetic diversity due to their maintenance of ancient polymorphisms or introgressions. RESULTS: We tested this hypothesis using the Iberian ibex (Capra pyrenaica) as an example. Previous studies based on large sample sizes taken from its principal populations have revealed that the Iberian ibex has a remarkably small MHC DRB1 diversity (only six remnant alleles) as a result of recent population bottlenecks and a marked demographic decline that has led to the extinction of two recognized subspecies. Extending on the geographic range to include non-studied isolated Iberian ibex populations, we sequenced a new MHC DRB1 in what seemed three small isolated populations in Southern Spain (n = 132). The findings indicate a higher genetic diversity than previously reported in this important gene. The newly discovered allele, MHC DRB1*7, is identical to one reported in the domestic goat C. aegagrus hircus. Whether or not this is the result of ancient polymorphisms maintained by balancing selection or, alternatively, introgressions from domestic goats through hybridization needs to be clarified in future studies. However, hybridization between Iberian ibex and domestic goats has been reported in Spain and the fact that the newly discovered allele is only present in one of the small isolated populations and not in the others suggests introgression. The new discovered allele is not expected to increase fitness in C. pyrenaica since it generates the same protein as the existing MHC DRB1*6. Analysis of a microsatellite locus (OLADRB1) near the new MHC DRB1*7 gene reveals a linkage disequilibrium between these two loci. The allele OLADRB1, 187 bp in length, was unambiguously linked to the MHC DRB1*7 allele. This enabled us to perform a DRB-STR matching method for the recently discovered MHC allele. CONCLUSIONS: This finding is critical for the conservation of the Iberian ibex since it directly affects the identification of the units of this species that should be managed and conserved separately (Evolutionarily Significant Units).

Characterizing the growth of Sarcoptes scabiei infrapopulations.

During the course of parasitic disease infestations, parasite population sizes change at both individual host (infrapopulation) and host population (metapopulation) levels. However, most studies only report epidemiological values for specific locations and times. In this study we analysed the dynamics of several Sarcoptes scabiei infrapopulations from experimentally infested Iberian ibex, Capra pyrenaica. We obtained mite counts by digesting small skin biopsies, which we compared with indices obtained from histopathological analyses performed on adjacent skin biopsies. We obtained the finite growth rate and the daily growth rate for the mite infrapopulations: mean ± SE = 11.53 ± 10.17 and 0.10 ± 0.08 mites/day, respectively. Mite counts derived from skin sample digestion did not correlate with the histological mite indices obtained from adjacent skin biopsies. At a metapopulational level, both indices of mite abundance were modelled using GLMMs and the factors influencing their variation are analysed and discussed. Our results suggest that mites are not distributed uniformly over the whole area of the skin lesion. Therefore, direct diagnoses of mange and mite counts could be inaccurate if only small skin samples are used.

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.

Acute phase proteins increase with sarcoptic mange status and severity in Iberian ibex (Capra pyrenaica, Schinz 1838).

Sarcoptic mange is a contagious skin disease caused by Sarcoptes scabiei, affecting both domestic and wild mammals, including the Iberian ibex (Capra pyrenaica), a medium-sized mountain ungulate almost endemic to the Iberian Peninsula. Acute phase proteins (APPs) could be an indicator of sarcoptic mange disease and severity in Iberian ibex. Serum samples from 131 healthy and sarcoptic mange-affected Iberian ibexes were collected from 2005 to 2012 in Sierra Nevada Natural Space in southern Spain. Serum alpha-1-acid glycoprotein (AGP), serum amyloid A (SAA) and haptoglobin (Hp) concentrations were quantified, and statistically significant differences according to sarcoptic mange disease and severity were assessed. Both AGP and SAA were significantly higher in the sarcoptic mange-affected ibexes than in the healthy ones as well as in the severely affected ibexes as compared to those with less than 50 % of the body surface affected. For the first time, changes in APP are reported in relation to sarcoptic mange in Iberian ibex. It is also reported for the first time that the intensity of APP increase depends on the severity of sarcoptic mange, which could be related with the pathological secondary amyloidosis, leading to organ dysfunction in severely mange-affected animals. Species and population differences in the increase of APP in response to sarcoptic mange could indicate individual and population differences in the immune capability of each population to deal with mange, population prevalence and mortality being the last indicators of such sensitivity.

Wildlife-Livestock Host Community Maintains Simultaneous Epidemiologic Cycles of Mycoplasma conjunctivae in a Mountain Ecosystem.

Infectious keratoconjunctivitis (IKC) is an eye disease caused by Mycoplasma conjunctivae that affects domestic and wild caprines, including Iberian ibex (Capra pyrenaica), a medium-sized mountain ungulate. However, its role in IKC dynamics in multi-host communities has been poorly studied. This study assessed M. conjunctivae in Iberian ibex and seasonally sympatric domestic small ruminants in the Natural Space of Sierra Nevada (NSSN), a mountain habitat in southern Spain. From 2015 to 2017, eye swabs were collected from 147 ibexes (46 subadults, 101 adults) and 169 adult domestic small ruminants (101 sheep, 68 goats). Mycoplasma conjunctivae was investigated through real-time qPCR and statistically assessed according to species, sex, age category, year, period, and area. The lppS gene of M. conjunctivae was sequenced and phylogenetically analysed. Mycoplasma conjunctivae was endemic and asymptomatic in the host community of the NSSN. Three genetic clusters were shared by ibex and livestock, and one was identified only in sheep, although each host species could maintain the infection independently. Naïve subadults maintained endemic infection in Iberian ibex, with an epizootic outbreak in 2017 when the infection spread to adults. Wild ungulates are epidemiologically key in maintaining and spreading IKC and other shared diseases among spatially segregated livestock flocks.

Home range in genus Capra: from polygons to Brownian bridges of scabietic and healthy Iberian ibexes (Capra pyrenaica).

Home range and home range overlap can be used to describe use of space and movement of wildlife. During the last years, advancements in technology have greatly improved our understanding of animal movement, especially among large herbivores. Wild ungulate abundance and distribution have increased in temperate areas. Moreover, their diseases-including sarcoptic mange in the Iberian Ibex (Capra pyrenaica)-have become a cause of concern for livestock, public health, and wildlife conservation. In this study, we first reviewed existing literature on the home range of species in the genus Capra. We then analyzed data from 52 GPS-GSM-collared Iberian ibexes, of which 33 were healthy and 19 were affected by sarcoptic mange from 3 different populations in the southeastern Iberian Peninsula to analyze: (1) differences in size and characteristics of home ranges obtained by the 3 most commonly used methodologies-minimum convex polygon, kernel density estimation, and Brownian bridges movement models (BBMMs); and (2) the impact of endemic sarcoptic mange on Iberian Ibex home range. The literature review revealed that available information on spatial behavior of Capra spp. was based only on 3 species, including the Iberian Ibex, estimated through a diversity of methods which made it difficult to compare results. We found positive correlations among the different home range estimation methods in the Iberian Ibex, with BBMMs proving to be the most accurate. This study is the first to use BBMMs for estimating home range in this species, and it revealed a marked seasonal behavior in spatial use, although sarcoptic mange smoothed such seasonal pattern. The seasonal overlaps obtained suggest that core areas of the Iberian Ibex change within wider home range areas, which are ecological parameters relevant to identifying key areas for species management and conservation.

Challenges of copro-parasitological surveys in wild Iberian ibex (Capra pyrenaica) populations addressed through a combination of molecular and statistical tools.

Copro-parasitological surveys in wildlife face challenges due to the secretive nature of many species and the unknown performance of the diagnostic tests employed. To overcome these issues, we used a combination of hierarchical models (site-occupancy and N-mixture models) applied to copro-parasitological data obtained from fecal samples assigned to the host species by molecular methods in the Iberian ibex in north-western Iberian Peninsula. The aims were to compare the performance of four diagnostic tests (Mini-FLOTAC, McMaster, Willis flotation, and natural sedimentation) and to use this methodological approach (molecular analysis with hierarchical models) to better estimate positivity proportion and shedding intensity in a wild ibex population. Pooled fecal samples were collected, and those confirmed by molecular analyses to be the host species in question were included in the study. Hierarchical models confirmed different performances of each diagnostic test, with Mini-FLOTAC showing higher sensitivity for eimeriid coccidia, Willis flotation (for proportion positive) and McMaster (for shedding intensity) in gastrointestinal Strongylida, and equal performance of MiniFlotac/Willis flotation (for proportion positive) and MiniFlotac/McMaster (for shedding intensity) in Moniezia spp. This study employed a combination of molecular and statistical methods that improved the estimates of prevalence and shedding intensity and allowed us to compare the performance of four diagnostic tests while assessing the effect of covariates. Such improvements are critical to enhancing inference in non-invasive wildlife copro-parasitological studies.

Co-infection patterns in the ectoparasitic community affecting the Iberian ibex Capra pyrenaica.

BACKGROUND: Sarcoptic mange is one of the main parasitic diseases affecting the Iberian ibex Capra pyrenaica. Scabietic animals suffer a decline in body condition and reproductive fitness and in severe cases may die. Although several previous studies of the pathology of this disease and the physiological changes it produces in ibex have been carried out in recent years, our knowledge of the relationship between Sarcoptes scabiei and other ectoparasites of this host is still limited. METHODS: We analysed 430 Iberian ibex skin samples. Ectoparasites were removed, counted and identified. Mite (S. scabiei) numbers were obtained after digesting the skin samples in a 5% KOH solution. We modelled mite numbers in terms of host sex and age, site, year, season and the presence of other ectoparasites such as ticks and lice using generalized linear mixed models (GLMMs) and ectoparasite co-occurrence patterns using two different models: the probabilistic model species co-occurrence and the generalized linear latent variable model (GLLVM). RESULTS: The ectoparasite community was mainly composed of S. scabiei, six ticks (Haemaphysalis sulcata, Haemaphysalis punctata, Rhipicephalus bursa, Rhipicephalus turanicus, Dermacentor marginatus and Ixodes ricinus) and two lice (Bovicola crassipes and Linognathus stenopsis). Adult male ibex harboured more mites than females. Mite numbers varied greatly spatially and seasonally and increased with the presence of other parasites. Some positive co-occurrence relationships between pairs of different ectoparasites were observed, particularly between ticks. The presence of S. scabiei negatively affected lice and H. sulcata numbers. CONCLUSIONS: Sarcoptic mange has spread above all in ibex populations in and around the Mediterranean Basin, where it is now found in almost a third of its host's range. Mite numbers varied seasonally and spatially and were higher in male hosts. The presence of S. scabiei had a negative effect on lice numbers but favoured the presence of ticks.

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.

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.

Remote mapping of foodscapes using sUAS and a low cost BG-NIR sensor.

The assessment of landscape condition for large herbivores, also known as foodscapes, is fast gaining interest in conservation and landscape management programs worldwide. Although traditional approaches are now being replaced by satellite imagery, several technical issues still need to be addressed before full standardization of remote sensing methods for these purposes. We present a low-cost method, based on the use of a modified blue/green/near-infrared (BG-NIR) camera housed on a small-Unmanned Aircraft System (sUAS), to create foodscapes for a generalist Mediterranean ungulate: the Iberian Ibex (Capra pyrenaica) in Northeast Spain. Faecal cuticle micro-histological analyses were used to assess the dietary preferences of ibexes and then individuals of the most common plant species (n = 19) were georeferenced to use as test samples. Because of the seasonal pattern in vegetation activity, based on the NDVI (Smooth term Month = 21.5, p-value < .01, R2 = 43%, from a GAM), images were recorded in winter and spring to represent contrasting vegetation phenology using two flight heights above ground level (30 and 60 m). Additionally, the range of image pixel sizes was 3.5-30 cm with the smallest pixel size representing the highest resolution. Boosted Trees were used to classify plant taxa based on spectral reflectance and create a foodscape of the study area. The number of target species, the sampling season, the height of flight and the image resolution were analysed to determine the accuracy of mapping the foodscape. The highest classification error (70.66%) was present when classifying all plant species using a 30 cm pixel size from acquisitions at 30 m height. The lowest error (18.7%), however, was present when predicting plants preferred by ibexes, at 3.5 cm pixel size acquired at 60 m height. This methodology can help to successfully monitor food availability and seasonality and to identify individual species.

Spatiotemporal monitoring of selected pathogens in Iberian ibex (Capra pyrenaica).

An epidemiological surveillance programme was carried out to assess exposure and spatiotemporal patterns of selected pathogens (Brucella spp., Mycobacterium avium subsp. paratuberculosis (MAP), Mycoplasma agalactiae, Pestivirus and bluetongue virus (BTV)) in Iberian ibex (Capra pyrenaica) from Andalusia (southern Spain), the region with the largest population of this species. A total of 602 animals in five distribution areas were sampled during 2010-2012 (P1) and 2013-2015 (P2). The Rose Bengal test (RBT) and complement fixation test (CFT) were used in parallel to detect anti-Brucella spp. antibodies. Commercial ELISAs were used to test for antibodies against the other selected pathogens. Sera positive for BTV and Pestivirus by ELISA were tested by serum neutralization test (SNT) to identify circulating serotypes/genotypes. The overall seroprevalences were as follows: 0.4% for Brucella spp. (2/549; CI 95%: 0.1-1.3) (14/555 positive by RBT; 2/564 by CFT), 0.5% for MAP (3/564; CI 95%: 0.1-1.5), 5.7% for M. agalactiae (30/529; CI 95%: 3.9-8.0), 11.1% for Pestivirus (58/525; CI 95%: 8.5-14.1) and 3.3% for BTV (18/538; CI 95%: 2.0-5.2). Significantly higher seropositivity to both M. agalactiae and BTV was observed in P1 compared with P2. Spatiotemporal clusters of high seroprevalence were also found for M. agalactiae in four of the five sampling areas in 2010, and for BTV in one of five areas in 2012. Specific antibodies against BTV-4, BDV-4 and BVDV-1 were confirmed by SNT. Our results indicate that the Iberian ibex may be considered spillover hosts of Brucella spp. and MAP rather than true reservoirs. The prevalence of antibodies against M. agalactiae and BTV suggests spatiotemporal variation in the circulation of these pathogens, while Pestivirus has a moderately endemic circulation in Iberian ibex populations. Our study highlights the importance of long-term surveillance for a better understanding of the spatiotemporal distribution of shared infectious diseases and providing valuable information to improve control measures at the wildlife-livestock interface.

Method validation, reference values, and characterization of acute-phase protein responses to experimentally induced inflammation and bluetongue virus infection in the Iberian ibex.

BACKGROUND: Acute phase protein (APP) concentrations can change due to inflammation and be used to monitor disease in the Iberian ibex (Capra pyrenaica). OBJECTIVES: This study aimed to validate Haptoglobin (Hp) and serum amyloid A (SAA) analytes, establish reference values, and characterize Hp and SAA responses in the Iberian ibex after experimentally induced inflammation and experimental bluetongue virus (BTV) infection. METHODS: Sera from 40 free-ranging box-trapped ibexes were used to establish Hp and SAA reference values. Six healthy ibexes were subcutaneously injected with 5 mL of turpentine, then, blood samples were taken, and clinical evaluations were performed on days 0, 1, 2, 3, 4, 7, and 14 postinjection. Another seven ibexes were challenged with BTV. Serum Hp and SAA concentrations were quantified using commercial assays following the manufacturer's instructions. RESULTS: Intra-assay precision and linearity were acceptable for both Hp and SAA. Intra-assay variation for high and low concentration of Hp and SAA were 9.74% and 17.31% and 16.49% and 12.89%, respectively. Inter-assay variation was higher for the low APP concentrations. Reference values for the healthy Iberian ibexes were (median, minimum, and maximum values) 0.2 (0.12-0.64) g/L for Hp and 4.74 (0.05-29.54) mg/L for SAA. Both Hp and SAA acted as a moderate and a major APP, respectively, and each could distinguish animals with turpentine-induced inflammation from those without. Hp and SAA did not change in asymptomatic BTV-infected animals. CONCLUSION: This study validated Hp and SAA analytes and provided basal reference values for these analytes in the Iberian ibex. Both APPs were able to discriminate between healthy and diseased Iberian ibexes animals during turpentine-induced inflammatory processes.

How sensitive and specific is the visual diagnosis of sarcoptic mange in free-ranging Iberian ibexes?

BACKGROUND: Sarcoptic mange is a broadly distributed parasitic disease caused by Sarcoptes scabiei that affects wild mammals from all over the world, including the Iberian ibex (Capra pyrenaica). Selective culling of the scabietic individuals is the main management measure for disease control in Iberian ibex populations. Although visual identification of mange-compatible lesions is the reference method to decide the target individual, both false negative and positive cases are common in the wild. The aim of this work is to determine the sensitivity (SE), and the specificity (SP) of selective culling after evaluating 403 ibexes hunted in the Sierra Nevada Nature Space for sarcoptic mange control between 2002 and 2015. METHODS: A combination of skin scrapings and potassium hidroxide (KOH) skin digestion was used for sarcoptic mange diagnosis. Generalized linear models (GLM) were used to assess the effects of sex, age (juveniles and adults) and period of the year (wet and dry periods) on the SE and SP of the visual diagnosis method. RESULTS: The SE obtained for the visual determination of scabietic ibexes was 87.14%, whereas the SP was 60.71%. According to our model selection, SE of the visual diagnosis was explained by the additive effects of age and the period of the year. In fact, SE was lower in juveniles (64.76%) than in adults (94.26%) and during the dry period (73.44%) as compared to the wet period (92.09%). On the other hand, SP was best explained by the GLM including the additive effects of sex and the period of the year. The visual diagnosis of sarcoptic mange resulted less specific in females (22.73%) than in males (74.19%) and during the wet (55.22%) than in the dry period (82.35%). CONCLUSIONS: Maximizing SE and SP is essential to achieving a high rate of removal of affected individuals from the environment without eliminating potentially resistant individuals. Selective culling must be conservative during the wet period and with females due to the lower SP. Conversely, visual diagnosis of scabietic juveniles and during the dry period has to be improved, due to the lower SE.

On the possible role of ticks in the eco-epidemiology of Coxiella burnetii in a Mediterranean ecosystem.

Ruminant livestock is the main reservoir of Coxiella burnetii (Cb), but little is known about the role of wildlife and ticks in its epidemiology. The Iberian ibex (Capra pyrenaica, Schinz 1838) population of "Ports de Tortosa i Beseit" (NE Spain) suffers intense tick infestations and low reproduction rates. This study aims to (1) assess the relationship between infection in ibexes (detection of serum antibodies and/or of Cb DNA in tissues) and Cb DNA presence in ticks hosted by the same ibexes; and (2) identify Cb associated risk factors. Between 2011 and 2015, serum (n = 130), spleen (n = 72), lymph node (n = 89) and tick (n = 669) samples from 134 hunter-harvested ibexes were collected. Antibody detection was performed by ELISA and Cb DNA presence was assessed by PCR. Potential risk factors were assessed with regression tree models. Although 30% of the ibexes (39/130; 95%CI, [10%-29.8%]) had antibodies, Cb DNA was detected in only 9.8% of the ibexes (11/112; 95%CI [7.6%-27.25%]). The prevalence of Cb-carrier ticks averaged 10% and exceeded 20% for the genus Haemaphysalis. However, lacking correlation between infection in ibexes and their ticks does not support tick-to-ibex transmission or vice versa. Tree modelling points to host, population and environmental factors as drivers of Cb infection in ticks and suggests connections with the domestic cycle. The percentage of Cb-carrier ticks detected is noteworthy. Along with heavy tick infestations, it suggests vector potential for these tick species, especially for the genera Rhipicephalus and Haemaphysalis. Since vector competence has not been assessed in these tick species, a classic vector role cannot be proposed nor discarded, but promoter factors of vector capacity occur. In addition, the risk of tick-borne infection through tick excreta should not be neglected. While the airborne route is the preeminent route for Cb infection, ticks' contribution to Cb epidemiology deserves further attention.

HPLC-QTOF method for quantifying 11-ketoetiocholanolone, a cortisol metabolite, in ruminants' feces: Optimization and validation.

Studies of animal ecology can benefit from a quantified understanding of eco-physiological processes and, in particular, of the physiological responses in free-ranging animals to potential stressors. The determination of fecal cortisol metabolites as a noninvasive method for monitoring stress has proved to be a powerful tool. High-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) has emerged as the most accurate method for avoiding problems related to the nonspecificity of immunoassays. In this study, we optimize and validate a reliable method using HPLC-MS/MS for quantifying 11-ketoetiocholanolone (11-k), a representative fecal cortisol metabolite in ruminants. An appropriate extraction and purification procedure was developed taking into account the complex nature of feces. The final extract obtained was then analyzed with HPLC-MS/MS using a quadrupole-time-of-fly (QTOF) tandem mass spectrometer with an electrospray ionization interface operating in positive mode, which allowed an unequivocal determination of the metabolite due to its accurate mass capabilities. After rigorous optimization of both sample extraction and the HPLC-QTOF parameters, making use of feces from free-ranging Iberian ibex, ideal conditions were established. Matrix-matched standards were used to calibrate the method. The limit of detection and quantification was 13- and 40- ng/g, respectively. The validation of the method was performed with recoveries in the range of 85-110%, a figure much higher than the 60% obtained with the previous extraction methods used in our laboratory, and with relative standard deviations (RSDs) no higher than 15% for the complete analytical procedure, including extraction and analysis. The time required for the fecal 11-k analysis was greatly reduced in comparison with the previous work carried out in our laboratory. This is the first time that QTOF mass detection coupled with HPLC has been validated for 11-k quantification in feces from free-ranging ruminants such as Iberian ibex. Given the high selectivity and sensitivity attained, our method could become a useful tool for noninvasive stress quantification in ruminants.

Endemic caseous lymphadenitis in a wild Caprinae population.

Between 2010 and 2013, an outbreak of caseous lymphadenitis (CLA) occurred in a captive stock of Iberian ibexes (Capra pyrenaica, Schinz 1838) maintained for conservation purposes in the National Game Reserve 'Ports de Tortosa i Beseit' (PTB), northeastern Spain. The aim of this study was to assess the CLA status in the free-ranging donor population of ibexes. An ELISA test to detect antibodies to CLA was performed in 360 hunter-harvested ibexes between 2007 and 2013. A spatial analysis and recursive partitioning approaches with regression tree models were used to explore CLA-associated risk factors. Nineteen per cent of ibexes had antibodies to CLA. Significant differences in seroprevalence were observed among game management areas but no clear trends of CLA occurrence were observed over the study period. Ibexes from areas of higher population density and living close to aggregation points displayed a higher probability of testing positive to CLA. These results suggest that CLA is endemic in the Iberian ibex population inhabiting PTB and density-dependent risk factors. To the best of our knowledge, this is the first record of CLA maintenance in a free-ranging wild Caprinae population.