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Influenza A viruses ( IAVs) are negative-sense, single-stranded and segmented RNA viruses of the Orthomyxoviridae family that may cause acute respiratory disease in a wide range of birds and mammals. Susceptibility of several species within the family Mustelidae to IAVs has been reported as a result of natural or experimental infections. The objectives of this study were to assess whether free-ranging American mink populations from Northern Spain were infected with IAV and try to define the role of this species in the epidemiology of IAV. Sera from 689 American mink from Northern Spain captured between 2011 and 2014 were tested for the presence of antibodies against IAVs using a commercial competition cELISA. Positive sera were further analysed with haemagglutination inhibition ( HI) assay. Fifteen of the 689 (2.2%, 1.3-3.6 CI95%) of the American minks analysed were ELISA positive. No significant differences were observed between years of capture, provinces, river basins, sexes or ages of the animals. All seropositive sera resulted negative to the panel strains used in the HI assay, showing that the most relevant strains circulating in swine, the most relevant avian subtypes (H5 and H7) and the H10N4 subtype isolated in minks have not been circulating in this free-ranging exotic carnivore from Spain. In the light of these results, the free-range American mink from Northern Spain do not seem to have an important role in the epidemiology of IAVs. [ABSTRACT FROM AUTHOR]

Abstract The bladderworm Pearsonema (syn Capillaria) plica affects domestic dogs and wild carnivores worldwide. A high prevalence in red foxes (Vulpes vulpes) has been reported in many European countries. P. plica inhabits the lower urinary tract and is considered to be of low pathogenic significance in dogs mostly causing asymptomatic infections. However, a higher level of pathogenicity has been reported in foxes. A severe cystitis associated with numerous bladderworms was found in a captive arctic fox (Vulpes lagopus) originating from the endangered Fennoscandian arctic fox population. To our knowledge this is the first description of P. plica infection in an arctic fox.

Identifying the role that host species play in pathogen transmission and maintenance is crucial for disease control, but it is a difficult task, in particular for vector-borne and multi-host pathogens, and especially when wildlife species are involved. This is the case for a Crimean-Congo hemorrhagic fever virus (CCHFV) hotspot in north-eastern Spain, where Iberian ibex ( Capra pyrenaica ) and wild boar ( Sus scrofa ) are involved, but their roles in disease transmission are unclear. In this context, we studied the dynamics of CCHFV transmission in these two species during the collapse of an Iberian ibex population due to a sarcoptic mange outbreak. We carried out a repeated cross-sectional study measuring the trends of CCHFV seroprevalence in Iberian ibex and wild boar and their abundances. In addition, we identified the tick species present in this area on the vegetation and on wild boars, and evaluated relevant meteorological factors. Results show that while the trends in CCHFV seroprevalence in Iberian Ibex and density of wild boars remained constant ( p  = 1.0 and p  = 0.8, respectively), both the trends in Iberian ibex census and CCHFV seroprevalence in wild boars decreased significantly ( p  = 0.003 and p  = 0.0001, respectively), and were correlated (Spearman's rank, 0.02 <  p -adjusted<0.05). The correlation between the patterns of reduction of Iberian ibex abundance and the decrease of seroprevalence in wild boars suggests some sort of shared transmission cycle between the two species. Data from tick species in the area suggest a possible role of Rhipicephalus bursa in CCHFV transmission. The dynamics of CCHFV were unlikely caused by changes in meteorological variables such as temperature or water vapor pressure deficit. Further studies will be needed to confirm these hypotheses., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Alpine grasslands are essential for carbon sequestration and food supply for domestic and wild herbivores inhabiting mountainous areas worldwide. These biomes, however, are alternatively threatened by the abandonment of agricultural and livestock practices leading to a fast-growing shrubification process while other mountain grasslands are suffering from the impacts of overgrazing. The functioning of alpine meadow ecosystems is primarily driven by climatic conditions, land-use legacies and grazing. However, although it is critically important, the role of large herbivores on the aboveground biomass and protein content of palatable plants is poorly understood for most alpine meadows. In this work, we explore the effects of grazing on grassland vegetation at two different spatial and temporal scales in the Eastern Pyrenees, Spain. Remote sensing was used to assess the effect of high and moderate grazing (HG and MG respectively) on grass biomass using the leaf area index (LAI) at the meso-scale (patches between 2.3 and 38.7 ha). We also explored the impact of null (NG), overgrazing (MO, mimicked overgrazing) and high (HG) grazing intensities at local scale setting eighteen 1 m 2 exclusion boxes in six meadows (three boxes each) commonly used by domestic and wild ungulates. Historical satellite data showed that LAI values are greater in high than in low grazed areas (HG, mean = 0.66, LG, mean = 0.55). Along the same lines, high and moderate grazing pressures improved biomass production at the local-scale (HG, mean = 590.3 g/m 2 , MO, mean = 389.3 g/m 2 and NG, mean = 110.8 g/m 2 ). Crude protein content reached higher values under MO pressure than under HG pressure. Our results confirm that grazing intensity exerts significant changes on the above-ground biomass production and the protein content of plants consumed by domestic (cattle and horses) and wild ungulates (Southern Chamois, Rupicapra pyrenaica). We can conclude that ungulates sustain biomass and nutritive values of grass exerting a negligible effect on biomass and protein content of woody vegetation. Our results will inform management guidelines to support profitable grazing activities and promote conservation of the open landscapes in the alpine ecosystems under the current global change scenario., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier B.V.)

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., (© 2020 Wiley-VCH GmbH.)

Antimicrobial resistance (AMR) has spread worldwide due to the inappropriate use of antimicrobial drugs in human and veterinary medicine, becoming a public health problem. However, little is known about its occurrence and maintenance in wild animals, and very few studies have been carried out in ecosystems subjected to low human pressure. In our study, nasal and lung swabs were collected from hunted Pyrenean chamois ( Rupicapra pyrenaica ), and nasal swabs from sympatric domestic sheep were also collected. The swabs were cultured in agar plates to obtain bacterial isolates from the Pasteurellaceae family. The presence of AMR was assessed in a total of 28 Pasteurellaceae isolates from 45 Pyrenean chamois, and 9 isolates from sympatric domestic sheep found in the National Hunting Reserve of Freser-Setcases (Northeastern Pyrenees, Spain). The isolates belonged to one of the following three species: Pasteurella multocida, Mannheimia haemolytica and Bibersteinia trehalosi. Some P. multocida and M. haemolytica isolates tested positive for AMR. The statistical analysis revealed no differences between the AMR levels from chamois and domestic sheep isolates. However, one P. multocida of chamois origin presented resistance to cephalosporins and fluoroquinolones, which are antibiotics of critical importance for human health. Further studies are required to elucidate potential routes of dissemination of AMR genes in natural environments and assess any significant persistence in wildlife to design risk mitigation actions.

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.

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., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Since 2001, Pyrenean chamois (Rupicapra pyrenaica pyrenaica) populations have been affected by border disease virus (BDV) causing mortalities of more than 80% in some areas. Field studies carried out in France, Andorra, and Spain have shown different epidemiological scenarios in chamois populations. This study was designed to confirm the presence of BDV strains of a high and low virulence in free-ranging chamois populations from Pyrenees and to understand the implications of these findings to the diverse epidemiological scenarios. An experimental infection of Pyrenean chamois with a high-virulence (Cadí-6) and low-virulence (Freser-5) BDV strains was performed. Pregnant and non-pregnant animals with and without antibodies against BDV were included in each group. Cadí-6 BDV strain was confirmed to be of high virulence for seronegative adults and their foetuses. The antibody negative chamois infected with Freser-5 BDV strain did not show symptoms, presented less viral distribution and RNA load in tissues than Cadí-6 group, and cleared the virus from the serum. However, foetuses died before the end of the experiment and RNA virus was detected in sera and tissues although with lower RNA load than the Cadí-6 group. Chamois from both groups presented lesions in brain but the ones infected with the low-virulence Freser-5 BDV strain were mild and most likely transient. In both groups, seropositive pregnant females and all but one of their foetuses did not present viraemia or viral RNA in tissues. The existence of a low-virulence strain has been confirmed experimentally and related to chamois population infection dynamics in the area where it was isolated. Such strain may persist in the chamois population through PI animals and may induce cross-protection in chamois against high-virulence strains. This study demonstrates that viral strain diversity is a significant factor in the heterogeneity of epidemiological scenarios in Pyrenean chamois populations., (© 2019 Blackwell Verlag GmbH.)

In alpine habitats, the seasonally marked climatic conditions generate seasonal and spatial differences in forage availability for herbivores. Vegetation availability and quality during the growing season are known to drive life history traits of mountain ungulates. However, little effort has been made to understand the association between plant phenology and changes in the foraging strategies of these mountain dwellers. Furthermore, this link can be affected by the seasonal presence of livestock in the same meadows. The objective of this work was to study the seasonal changes in diet composition of Pyrenean chamois (Rupicapra p. pyrenaica) and its relationship to primary production trends in a Mediterranean alpine environment. Moreover, diet composition in two populations with contrasting livestock pressure was compared in order to study the effect of sheep flocks on the feeding behaviour of chamois. From 2009 to 2012, monthly diet composition was estimated by cuticle microhistological analysis of chamois faeces collected in the eastern Pyrenees. The primary production cycle was assessed by remote sensing, using the Normalized Difference Vegetation Index. Additionally, the diet of sheep sharing seasonally the subalpine and alpine meadows with chamois was analysed. Diet selection of chamois and sheep and their overlap was also assessed. Our results show an intra-annual variation in the diet composition of Pyrenean chamois and demonstrate a strong relationship between plant consumption dynamics and phenology in alpine areas. In addition, Calluna vulgaris, Cytisus spp. and Festuca spp., as well as forbs in the summer, are found to be key forage species for Pyrenean chamois. Furthermore, this study couldn't detect differences between both chamois populations despite the presence of sheep flocks in only one area. However, the detection of a shift in the diet of chamois in both areas after the arrival of high densities of multi-specific livestock suggest a general livestock effect. In conclusion, Pyrenean chamois are well adapted to the variations in the seasonal availability of plants in alpine habitats but could be disturbed by the seasonal presence of livestock. Due to the key plants in their diet, we suggest that population management programmes should focus on the preservation of mixed grasslands composed of patches of shrubs and herbs. The effects of climate change and shrub expansion should be studied as they may potentially affect chamois population dynamics through changes in habitat composition and temporal shifts in forage availability., Competing Interests: The authors have declared that no competing interests exist.

Water-borne transmission may play an important role in the epidemiology of Toxoplasma gondii. Mammals closely related to freshwater ecosystems, such as the American mink (Neovison vison), are potentially valuable sentinels for T. gondii. To assess the importance of freshwater ecosystems in T. gondii epidemiology, sera of 678 American minks collected during the 2010 to 2015 Spanish national eradication campaigns were tested for the presence of T. gondii antibodies using the modified agglutination test (MAT, cut-off 1:25). A high prevalence of samples, 78.8% (CI 95% : 75.5-81.8), were seropositive. In addition, a specific real-time PCR was performed in 120 brain samples and the parasite DNA was detected in 9.2% (CI 95% : 5.2-15.7). Significant differences in seroprevalence were detected among bioregions, with the highest levels detected in coastal areas, and by age. The higher seroprevalence observed in older animals (80.0% adults versus 68.7% juveniles) confirms the importance of the horizontal transmission. These results indicate a widespread presence of T. gondii oocysts in freshwater ecosystems from Spain and further support the importance of water-borne transmission in the epidemiology of T. gondii.

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.

Border Disease Virus (BDV) causes health and economic impact on livestock and is also of importance in wildlife conservation as it causes high mortality outbreaks in Pyrenean chamois (Rupicapra pyrenaica pyrenaica). Pastoral practices are known as a main interspecies pathogen transmission. Hence, the presence of pestivirus in transhumant sheep flocks and sympatric chamois was assessed in areas with different epidemiological scenarios of chamois BDV infections. Moreover, the present study had also the goal to identify if inter-specific infections occurred and when they happened. Five sheep flocks grazing in two alpine areas in the Pyrenees with two different BDV epidemiological scenarios in chamois populations were studied during two transhumant seasons. Sheep were sampled before and after transhumance. Pyrenean chamois sera and spleen samples from both areas where also studied during the same period. Antibodies against BDV were assessed by means of ELISA and VNT. A qRT-PCR was used in order to detect the virus. Seroprevalence in sheep ranged between 0 and 91.1% at the flock level. Chamois were found to have high seroprevalences (52.9-77.7%) in both areas, and four new BDV isolates were sequenced. One sheep farm presented persistent BDV circulation and three showed low BDV circulation. The after-transhumance period was identified as the moment when viral transmission occured in the first farm, associated to BDV strains of domestic origin, according to VNT results. However, the BDV isolate was genetical closely related to previous BDV strains from chamois origin. In another farm, antibodies in two of the three positive sera were associated to infection with a chamois-like BDV strain. Altogether indicates that occasional viral transmission from chamois to sheep may occur., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Wild boar populations are expanding throughout the world and intruding into periurban and urban areas. In the last years, wild boar has colonized several European cities, including our study area, the city of Barcelona. It is required to identify the main factors driving wild boar into urban areas prior to establish management measures. We built Boosted Regression Trees (BRT) using 3148 wild boar presences registered in the urban area of Barcelona from 2010 to 2014 to identify the variables correlated with these presences. The variables analysed included proxies for distance to source population, urban food resources, climate and urban habitat structure. Wild boars enter the urban area from close natural habitat using corridors such as streams, preferably in fragmented urban environment, looking for food such as urban green areas or dry pet food from cat colonies. Wild boar presence is higher in spring possibly due to the births of piglets and the dispersion of yearlings during that season, and also when natural resources in the Mediterranean habitat fail to satisfy the nutritional requirements of the wild boar population during the summer season. Management measures derived from this study are currently being applied in the city of Barcelona, including vegetation clearings in the wild boar entrance areas and an awareness campaign aimed at reducing the anthropogenic food availability for wild boars. The methodology used can be applied to other cities with wild boar or even other wildlife species issues. The comparison between the factors attracting wild boars into different urban areas would be helpful to understand the global phenomenon., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Functional roles of domestic and wild host populations in infectious keratoconjunctivitis (IKC) epidemiology have been extensively discussed claiming a domestic reservoir for the more susceptible wild hosts, however, based on limited data. With the aim to better assess IKC epidemiology in complex host-pathogen alpine systems, the long-term infectious dynamics and molecular epidemiology of Mycoplasma conjunctivae was investigated in all host populations from six study areas in the Pyrenees and one in the Cantabrian Mountains (Northern Spain). Detection of M. conjunctivae was performed by qPCR on 3600 eye swabs collected during seven years from hunted wild ungulates and sympatric domestic sheep (n = 1800 animals), and cluster analyses of the strains were performed including previous reported local strains. Mycoplasma conjunctivae was consistently detected in three Pyrenean chamois (Rupicapra p. pyrenaica) populations, as well as in sheep flocks (17.0% of sheep) and occasionally in mouflon (Ovis aries musimon) from the Pyrenees (22.2% in one year/area); statistically associated with ocular clinical signs only in chamois. Chamois populations showed different infection dynamics with low but steady prevalence (4.9%) and significant yearly fluctuations (0.0%- 40.0%). Persistence of specific M. conjunctivae strain clusters in wild host populations is demonstrated for six and nine years. Cross-species transmission between chamois and sheep and chamois and mouflon were also sporadically evidenced. Overall, independent M. conjunctivae sylvatic and domestic cycles occurred at the wildlife-livestock interface in the alpine ecosystems from the Pyrenees with sheep and chamois as the key host species for each cycle, and mouflon as a spill-over host. Host population characteristics and M. conjunctivae strains resulted in different epidemiological scenarios in chamois, ranging from the fading out of the mycoplasma to the epidemic and endemic long-term persistence. These findings highlight the capacity of M. conjunctivae to establish diverse interactions and persist in host populations, also with different transmission conditions.

Infectious keratoconjunctivitis (IKC) is a contagious eye disease primarily caused by Mycoplasma conjunctivae in domestic and wild Caprinae. Chlamydophila species have also been detected in ruminants with IKC. The objectives of this study are to investigate the ocular infection of M. conjunctivae and Chlamydiaceae and assess its interaction in relation to IKC in sheep and goats from remote communities around the Central Karakoram National Park in Pakistan, performing a combination of cross-sectional and case-control study design. Mostly asymptomatic and endemic infections of M. conjunctivae and Chlamydiaceae were found in sheep (19.3 per cent and 4.5 per cent, respectively) and goats (9.5 per cent and 1.9 per cent, respectively) from all communities, assessed by qPCR. Prevalence significantly differed between species only for M. conjunctivae (P=0.0184), which was also more prevalent in younger sheep (P<0.01). Chlamydophila pecorum was identified by sequencing and was related with IKC only when coinfection with M. conjunctivae occurred, which suggest a synergic interaction. Cluster analysis of M. conjunctivae strains revealed higher diversity of strains than expected, evidenced interspecific transmission and suggested a higher local livestock trade than previously assumed. These results highlight the widespread occurrence of M conjunctivae in sheep worldwide and its implications for wildlife should be assessed from a conservation perspective., Competing Interests: Competing interests: XFA was supported by the FI DGR programme from the Government of Catalonia., (© British Veterinary Association (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

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 [ C T ], 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. IMPORTANCE Mycoplasma 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., (Copyright © 2017 American Society for Microbiology.)

Optimal management of free-ranging herbivores requires the accurate assessment of an animal's nutritional status. For this purpose 'near-infrared reflectance spectroscopy' (NIRS) is very useful, especially when nutritional assessment is done through faecal indicators such as faecal nitrogen (FN). In order to perform an NIRS calibration, the default protocol recommends starting by generating an initial equation based on at least 50-75 samples from the given species. Although this protocol optimises prediction accuracy, it limits the use of NIRS with rare or endangered species where sample sizes are often small. To overcome this limitation we tested a single NIRS equation (i.e., multispecies calibration) to predict FN in herbivores. Firstly, we used five herbivore species with highly contrasting digestive physiologies to build monospecies and multispecies calibrations, namely horse, sheep, Pyrenean chamois, red deer and European rabbit. Secondly, the equation accuracy was evaluated by two procedures using: (1) an external validation with samples from the same species, which were not used in the calibration process; and (2) samples from different ungulate species, specifically Alpine ibex, domestic goat, European mouflon, roe deer and cattle. The multispecies equation was highly accurate in terms of the coefficient of determination for calibration R2 = 0.98, standard error of validation SECV = 0.10, standard error of external validation SEP = 0.12, ratio of performance to deviation RPD = 5.3, and range error of prediction RER = 28.4. The accuracy of the multispecies equation to predict other herbivore species was also satisfactory (R2 > 0.86, SEP < 0.27, RPD > 2.6, and RER > 8.1). Lastly, the agreement between multi- and monospecies calibrations was also confirmed by the Bland-Altman method. In conclusion, our single multispecies equation can be used as a reliable, cost-effective, easy and powerful analytical method to assess FN in a wide range of herbivore species.

Background: Even though male-biased parasitism is common in mammals, little effort has been made to evaluate whether higher parasitic burden in males results in an extra biological cost, and thus a decrease in fitness. Body condition impairment and the augmentation of oxidative stress can be used as indicators of the cost of parasite infections. Here, we examined relationships between gastrointestinal and respiratory helminths, body condition and oxidative stress markers (glutathione peroxidase, paraoxonase-1) in 28 Pyrenean chamois (Rupicapra p. pyrenaica) sampled in autumn., Results: Only male chamois showed a reduction in body condition and higher oxidative stress due to parasite infection, likely because of the extremely high parasite burdens observed in males., Conclusions: This study made evident a disparity in the physiological cost of multiple parasitism between sexes in a wild mammal, mainly due to parasitic richness. Because of the similar life expectancy in male and female chamois, we suggest that males may have developed natural mechanisms to compensate for higher parasite loads during the rut.

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.

Roads are widely recognized to represent a barrier to individual movements and, conversely, verges can act as potential corridors for the dispersal of many small mammals. Both barrier and corridor effects should generate a clear spatial pattern in genetic structure. Nevertheless, the effect of roads on the genetic structure of small mammal populations still remains unclear. In this study, we examine the barrier effect that different road types (4-lane highway, 2-lane roads and single-lane unpaved roads) may have on the population genetic structure of three species differing in relevant life history traits: southern water vole Arvicola sapidus, the Mediterranean pine vole Microtus duodecimcostatus and the Algerian mouse Mus spretus. We also examine the corridor effect of highway verges on the Mediterranean pine vole and the Algerian mouse. We analysed the population structure through pairwise estimates of FST among subpopulations bisected by roads, identified genetic clusters through Bayesian assignment approaches, and used simple and partial Mantel tests to evaluate the relative barrier or corridor effect of roads. No strong evidences were found for an effect of roads on population structure of these three species. The barrier effect of roads seems to be site-specific and no corridor effect of verges was found for the pine vole and Algerian mouse populations. The lack of consistent results among species and for each road type lead us to believe that the ability of individual dispersers to use those crossing structures or the habitat quality in the highway verges may have a relatively higher influence on gene flow among populations than the presence of crossing structures per se. Further research should include microhabitat analysis and the estimates of species abundance to understand the mechanisms that underlie the genetic structure observed at some sites., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Neospora caninum infection is a leading cause of abortion in cattle worldwide. The pathogenesis of bovine neosporosis, particularly during the second term of gestation when most abortions occur in naturally infected dams, is poorly understood. In the present study foetal death was observed in 3 of 6 experimentally infected dams at 110 days of gestation after 6 weeks of experimental period. All experimental heifers were febrile between 3 and 5 days post infection (dpi). Inoculated dams seroconverted by 3-4 weeks post-infection with higher mean antibody titres in aborting dams compared to non-aborting heifers, although not significantly (p > 0.05). Neospora caninum DNA was detected in all infected foetuses and placentas, and three infected foetuses also had N. caninum antibodies. The parasite burden was higher in the brain of dead/aborted foetuses than in live foetuses. Interestingly, high IFN-γ production was detected in foetal fluids of a dead foetus found upon euthanasia of its dam, while no IFN-γ was observed in amniotic, allantoic and/or foetal fluids in the three infected foetuses that were alive upon maternal euthanasia. The present study confirms that the infection of dams on gestation day 110 with 10(7) tachyzoites of the Nc-Spain7 isolate causes abortion. The fact that some infected dams aborted and some did not is relevant to the understanding of N. caninum pathogenesis of abortion in naturally infected cows., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: Coxiella burnetii is a zoonotic bacterium that infects a wide range of animal species and causes the disease Q fever. Both wild and domestic ruminants may be relevant in the epidemiology of C. burnetii infection. In order to investigate the significance of the ruminant host community in the alpine and subalpine ecosystems of the Eastern Pyrenees, Northeastern Spain, in the epidemiology of Q fever, a serological survey was performed on samples from 599 wild and 353 sympatric domestic ruminants., Results: Specific antibodies against C. burnetii were detected with a commercial enzyme-linked immunosorbent assay (ELISA). Domestic sheep showed the highest prevalence (12.7 %, CI 95 % 8.6-16.9), followed by European mouflon (Ovis orientalis musimon) with a 6.8 % prevalence (CI 95 % 1.6-12.1), red deer (Cervus elaphus) with 2.4 % (CI 95 % 0-5.6), and cattle with a prevalence of 1.1 % (CI 95 % 0-3.2). No positive domestic goats, fallow deer (Dama dama), roe deer (Capreolus capreolus) and Southern chamois (Rupicapra pyrenaica) were detected. Sheep flock prevalence was 75 % (nine of the 12 sheep flocks sampled were positive, within-flock prevalence ranging from 11.1 to 25.0 %), whereas cattle herd prevalence was 11.1 % (one out of the nine cattle herds sampled was positive, within-herd prevalence of 10.0 %., Conclusions: Both domestic and wild ruminants from the alpine and subalpine ecosystems of the Eastern Pyrenees were exposed to C. burnetii. The higher seroprevalence in sheep and its relative abundance suggest that this species may have a major contribution to the ecology of C. burnetii. Conversely, wild ruminants do not seem to represent a relevant host community for C. burnetii maintenance in the Eastern Pyrenees.

Understanding the spread of Toxoplasma gondii (T. gondii) in wild birds, particularly in those with opportunistic feeding behavior, is of interest for elucidating the epidemiological involvement of these birds in the maintenance and dissemination of the parasite. Overall, from 2009 to 2011, we collected sera from 525 seagull chicks (Yellow-legged gull (Larus michahellis) and Audouin's gull (L. audouinii)) from 6 breeding colonies in Spain and tested them using the modified agglutination test (MAT) for the presence of antibodies against T. gondii. Chick age was estimated from bill length. Main food source of seagull chicks was evaluated using stable isotope analyses from growing scapular feathers. Overall T. gondii seroprevalence was 21.0% (IC95% 17.5-24.4). A generalized linear mixed-effects model indicated that year (2009) and food source (freshwater) were risk factors associated to the individual risk of infection by T. gondii, while age (days) was close to significance. Freshwater food origin was related to the highest seroprevalence levels, followed by marine origin, supporting freshwater and sewages as important routes of dispersion of T. gondii. Year differences could indicate fluctuating rates of exposure of seagull chicks to T. gondii. Age ranged from 4 to 30 days and seropositivity tended to increase with age (P = 0.07), supporting that seropositivity is related to T. gondii infection rather than to maternal transfer of antibodies, which in gulls is known to sharply decrease with chick age. This study is the first to report T. gondii antibodies in Yellow-legged and Audouin's gulls, thereby extending the range of intermediate hosts for this parasite and underscoring the complexity of its epidemiology.

Though it is accepted that emerging infectious diseases are a threat to planet biodiversity, little information exists about their role as drivers of species extinction. Populations are also affected by natural catastrophes and other pathogens, making it difficult to estimate the particular impact of emerging infectious diseases. Border disease virus genogroup 4 (BDV-4) caused a previously unreported decrease in populations of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in Spain. Using a population viability analysis, we compared probabilities of extinction of a virtual chamois population affected by winter conditions, density dependence, keratoconjunctivitis, sarcoptic mange, and BD outbreaks. BD-affected populations showed double risk of becoming extinct in 50 years, confirming the exceptional ability of this virus to drive chamois populations.

The most widely used technique to assess helminth infection in both domestic and wild mammals is the faecal egg count (FEC). Most efforts to test the reliability of FEC as a proxy for parasite load are in small ruminant studies and limited work has evaluated the use of FEC in pigs. The aim of this study was to explore whether FEC is a reliable indicator of helminth load, and to evaluate the effects of sample storage on FEC accuracy in 59 wild boars. Though FEC was useful for assessing most helminth infections (e.g., Metastrongylus spp., Ascaris suum, Trichuris suis), stomach nematodes were often missed. The accuracy of FEC decreased over time, and thus it is recommended that samples be processed within 5 days of collection., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Pyrenean chamois (Rupicapra pyrenaica pyrenaica) is a nearly monomorphic mountain ungulate with an unbiased sex-specific overwinter adult survival. Few differences in gastrointestinal parasitism have been reported by coprology as yet. This study aims to assess diversity, prevalence, intensity of infection and aggregation of gastrointestinal nematodes in male and female adult chamois. We expect no differences in the parasite infection rates between sexes., Findings: Gastrointestinal tracts of 28 harvested Pyrenean chamois in the Catalan Pyrenees (autumn 2012 and 2013) were necropsied and sexual differences in the diversity and structure of parasite community, prevalence, intensity of infection, and richness were investigated. We found 25 helminth species belonging to 13 different genera., Conclusions: Contrary to our expectations, male chamois showed different parasite communities, higher prevalence, intensity of infection and richness than females. Such sexual differences were clear irrespective of age of individuals. Hence, male chamois must cope with a more diverse and abundant parasite community than females, without apparent biological cost. Further research will be required to confirm this hypothesis.

Background: Infectious keratoconjunctivitis (IKC) is a clinical condition affecting eyes of domestic and wild Caprinae worldwide, and Mycoplasma conjunctivae is considered the primary causative agent of IKC in sheep, goats and wild Caprinae. Domestic ruminants from high mountain habitats share grazing areas with wild mountain ungulates, such as chamois (Rupicapra spp.), Alpine ibex (Capra ibex) and European mouflon (Ovis aries musimon), and domestic sheep seem to act as M. conjunctivae reservoir. In this study, the presence of M. conjunctivae in domestic sheep and goats from the two main mountain ranges of Northern Spain, the Pyrenees and the Cantabrian Mountains, has been investigated., Results: Eye swabs were obtained from 439 domestic small ruminants selected from flocks that seasonally graze in alpine meadows during three consecutive years (2011-2012-2013). Seventy-nine out of the 378 domestic sheep (20.9%) tested positive to a M. conjunctivae specific real time-PCR (rt-PCR) in at least one eye, whereas all the 61 sampled domestic goats were negative. Statistically significant higher prevalence and higher proportion of infected flocks (P < 0.001) was observed in the Pyrenees (25.7%; 12 flocks out of 13), where M. conjunctivae is widespread and probably endemic in domestic sheep, than in the Cantabrian Mountains (7.8%; one flock out of six). Twenty-five sheep (three from the Pyrenees and 22 from the Cantabrian Mountains) which showed clinical signs consistent with infectious keratoconjunctivitis (IKC) were negative by rt-PCR. In contrast, 62 out of the 71 (87.3%) M. conjunctivae-positive sheep from the Pyrenees and the eight positive sheep from the Cantabrian Mountains were asymptomatic., Conclusions: This study provides rt-PCR-based evidences of M. conjunctivae maintenance in domestic sheep, as well as a relationship between prevalence in domestic sheep and previously reported M. conjunctivae and IKC in wild ruminants. Domestic goats do not seem to play an important role in the epidemiology of M. conjunctivae in alpine habitats from Northern Spain.

The bladderworm Pearsonema (syn Capillaria) plica affects domestic dogs and wild carnivores worldwide. A high prevalence in red foxes (Vulpes vulpes) has been reported in many European countries. P. plica inhabits the lower urinary tract and is considered to be of low pathogenic significance in dogs mostly causing asymptomatic infections. However, a higher level of pathogenicity has been reported in foxes. A severe cystitis associated with numerous bladderworms was found in a captive arctic fox (Vulpes lagopus) originating from the endangered Fennoscandian arctic fox population. To our knowledge this is the first description of P. plica infection in an arctic fox.