Projection of current and future distribution of adaptive genetic units in an alpine ungulate.

Climate projections predict major changes in alpine environments by the end of the 21st century. To avoid climate-induced maladaptation and extinction, many animal populations will either need to move to more suitable habitats or adapt in situ to novel conditions. Since populations of a species exhibit genetic variation related to local adaptation, it is important to incorporate this variation into predictive models to help assess the ability of the species to survive climate change. Here, we evaluate how the adaptive genetic variation of a mountain ungulate-the Northern chamois (Rupicapra rupicapra)-could be impacted by future global warming. Based on genotype-environment association analyses of 429 chamois using a ddRAD sequencing approach, we identified genetic variation associated with climatic gradients across the European Alps. We then delineated adaptive genetic units and projected the optimal distribution of these adaptive groups in the future. Our results suggest the presence of local adaptation to climate in Northern chamois with similar genetic adaptive responses in geographically distant but climatically similar populations. Furthermore, our results predict that future climatic changes will modify the Northern chamois adaptive landscape considerably, with various degrees of maladaptation risk.

Age-specific survival of territorial and non-territorial male chamois.

How alternative reproductive tactics (ARTs) are maintained in wildlife populations is one of the major questions in evolutionary biology. As a dominant status, territoriality is typically linked to increased mating opportunities, and one explanation why this behaviour coexists with other tactics is that dominance implies survival costs. Such a trade-off may occur in the Northern chamois Rupicapra rupicapra, as reproductive advantages of territorial males over non-territorial males could be counterbalanced by a reduction in survival mediated through energy expenditure, stress and parasitic infections, ultimately favouring ART coexistence. Here, we analysed age-dependent survival probabilities of territorial (n = 15) and non-territorial (n = 16) adult chamois using information collected over 12 years between 2010 and 2021 in the Gran Paradiso National Park (Western Italian Alps). Survival rates were estimated with a CMR approach using Burnham's joint modelling of live encounter and dead recovery data. The model selection procedure, based on AICc value minimisation, supported a linear decrease of survival with age but the results did not match our predictions, as territorial chamois did not have lower survival rates than non-territorial chamois. In contrast, territorial males appeared to enjoy reproductive success at lower survival costs. This, in turn, supports the role of other factors, such as snow-dependent environmental stochasticity, in the maintenance of ARTs in chamois populations. The limited sample size, however, calls for caution in interpretation, and long-term studies of lifetime reproductive success and survival are necessary to clarify the mechanisms underlying the expression and coexistence of different reproductive behaviours in this species.

Immunolocalisation of aquaporins 3, 7, 9 and 10 in the epididymis of three wild ruminant species (Iberian ibex, mouflon and chamois) and sperm cryoresistance.

CONTEXT: In the epididymis, epithelial cells manage changes in the luminal environment for proper sperm maturation. Moreover, aquaglyceroporins, a subgroup of aquaporins (AQP), modulate the transport of water, glycerol and other small molecules in epithelial cells. AIMS: We aim to characterise the lining epithelium, quantify its cell composition and immunolocalise the aquaglyceroporins AQP3, AQP7, AQP9 and AQP10 alongside the epididymal ductus of three wild ruminant species, and to determine if species-specific differences could be associated with cauda sperm cryoresistance variations. METHODS: Epididymides from Iberian ibex (n =5), mouflon (n =5) and chamois (n =6) were obtained. Cauda spermatozoa were collected and sperm parameters were analysed before and after freezing. Histology and immunohistochemistry of AQP3, 7, 9, 10 and T-CD3 were performed in the caput, corpus and cauda epididymal regions. KEY RESULTS: This work first describes the lining epithelium in Iberian ibex, mouflon and chamois epididymis along the three anatomical regions, consisting of principal, basal, apical, clear and halo cells. However, the percentage of each cell type differed in ibex compared to mouflon and chamois. The positive T-CD3 immunolabeling of all the halo cells confirmed their T-lymphocyte nature. Aquaglyceroporin expression patterns were similar among species, except for differences in AQP7 and AQP10 immunolocalisation in ibex. Species-specific differences in epididymal sperm cryoresistance were confirmed. CONCLUSIONS: The epididymal epithelium of the three wild ruminants differ in their relative number of cell types and AQP immunolocalisation, which ultimately appears to affect cauda epidydimal spermatozoa cryoresistance. IMPLICATIONS: Our study provides information on the relevance of the quantitative composition and AQP pattern expression in epididymal lining epithelium on sperm cryoresistance.

Comparative analysis of two genomes of Chlamydia pecorum isolates from an Alpine chamois and a water buffalo.

BACKGROUND: To date, whole genome sequencing has been performed mainly for isolates of Chlamydia trachomatis, C. pneumoniae, C. psittaci and C. abortus, but only a few isolates of C. pecorum have been entirely sequenced and this makes it difficult to understand its diversity and population structure. In this study the genome of two C. pecorum strains isolated from the lung of an Alpine chamois affected with pneumonia (isolate PV7855) and the brain of a water buffalo affected with meningoencephalomyelitis (isolate PV6959), were completely sequenced with MiSeq system (Illumina) and analyzed in their most polymorphic regions. RESULTS: The genome length and GC content of the two isolates were found to be consistent with other C. pecorum isolates and the gene content of polymorphic membrane proteins and plasticity zone was found to be very similar. Some differences were observed in the phospholipase genes for both isolates and in the number of genes in the plasticity zone, such as the presence of some hypothetical proteins in PV6959, not present in any other genomes analyzed in this study. Interestingly, PV6959 possesses an extra pmp and has an incomplete tryptophan biosynthesis operon. Plasmids were detected in both isolates. CONCLUSIONS: Genome sequencing of the two C. pecorum strains did not reveal differences in length and GC content despite the origin from different animal species with different clinical disease. In the plasticity zone, the differences in the genes pattern might be related to the onset of specific symptoms or infection of specific hosts. The absence of a tryptophan biosynthesis pathway in PV6959 may suggest a strict relationship between C. pecorum and its host.

Interacting lethal and nonlethal human activities shape complex risk tolerance behaviors in a mountain herbivore.

Animals perceive human activities as risky and generally respond with fear-induced proactive behaviors to buffer the circadian patterns of lethal and nonlethal disturbances, such as diel migrations (DMs) between risky places during safe nighttime and safer places during risky daytime. However, such responses potentially incur costs through movement or reduced foraging time, so individuals should adjust their tolerance when human activities are harmless, through habituation. Yet this is a challenging cognitive task when lethal and nonlethal risks co-occur, forming complex landscapes of fear. The consequences of this human-induced complexity have, however, rarely been assessed. We studied the individual DM dynamics of chamois (Rupicapra rupicapra rupicapra), 89 GPS-tracked individual-years, from/to trails in the French Alps in areas with co-occurring lethal (hunting) and nonlethal (hiking and skiing) disturbances, with different intensities across seasons. We developed a conceptual framework relying on the risk-disturbance hypothesis and habituation to predict tolerance adjustments of chamois under various disturbance contexts and across contrasted seasonal periods. Based on spatial and statistical analyses combining periodograms and multinomial logistic models, we found that DM in relation to distance to a trail was a consistent response by chamois (~85% of individuals) to avoid human disturbance during daytime, especially during the hiking and hunting periods. Such behavior revealed a low tolerance of most chamois to human activities, although there was considerable interindividual heterogeneity in DM. Interestingly, there was an increased tolerance among the most disturbed diel migrants, potentially through habituation, with chamois performing shorter DMs in areas highly disturbed by hikers. Crucially, chamois that were most human-habituated during the hiking period remained more tolerant in the subsequent harvesting period, which could increase their risk of being harvested. In contrast, individuals less tolerant to hiking performed longer DMs when hunting risk increased, and compared to hiking, hunting exacerbated the threshold distance to trails triggering DMs. No carryover effect of hunting beyond the hunting period was observed. In conclusion, complex human-induced landscapes of fear with co-occurring disturbances by nature-based tourism and hunting may shape unexpected patterns of tolerance to human activities, whereby animal tolerance could become potentially deleterious for individual survival.

Temporal stability of fecal cortisol metabolites in mountain-dwelling ungulates.

Fecal cortisol metabolites (FCMs) are widely used to track stress responses in wildlife and captive species. Rules of thumb suggest that samples should be collected as soon as possible after defecation, to avoid decay of FCMs. To date, however, only a few studies investigated the stability of defecated FCMs over time, and most of them were conducted in controlled laboratory conditions. Here, we investigated the stability of FCMs over seven consecutive days, in two mountain-dwelling ungulates, under natural environmental conditions using a semi-experimental approach. Fecal samples from Northern chamois Rupicapra rupicapra (n = 24) and red deer Cervus elaphus (n = 22) were collected in summer of 2020 within the Stelvio National Park, Italy, and placed in an open area above 2000 m a.s.l. For the next 7 days, we collected a portion of each sample, and all sub-samples were analyzed with an 11-oxoetiocholanolone enzyme immunoassay. Exposure, temperature, and precipitation were fitted as covariates in non-linear generalized mixed models to assess FCM variation over time, and competing models were selected using AICc. For chamois, the best model included only time as a predictor, while for red deer, it included time, precipitation, and exposure. For both species, FCM values decreased rapidly from the first days after deposition until the fourth day. For red deer, in northern-exposed samples, FCM values decreased slower than in south-exposed ones; furthermore, FCM values increased with increasing precipitation. Our results offer a solid methodological basis to wildlife researchers and practitioners interested in the investigation of the ecological factors affecting stress variation in wildlife and support the recommendation to collect samples as fresh as possible, to avoid misleading inference. Further studies are necessary to evaluate the stability of FCMs when other enzyme immunoassays are used.

Forests buffer the climate-induced decline of body mass in a mountain herbivore.

Climate change is known to affect key life-history traits, such as body mass, reproduction, and survival in many species. Animal populations inhabiting mountain habitats are adapted to extreme seasonal environmental conditions but are also expected to be especially vulnerable to climate change. Studies on mountain ungulates typically focus on populations or sections of populations living above the tree line, whereas populations inhabiting forested habitats are largely understudied. Here, we investigate whether forested areas can mitigate the impact of climatic change on life-history traits by evaluating the interactive effects of temperature and habitat characteristics on body mass variation in the Alpine chamois Rupicapra rupicapra rupicapra. We examined data of 20,573 yearling chamois collected from 1993 to 2019 in 28 mountain ranges in the Austrian Eastern Alps, characterized by different proportion of forest cover. Our results show that the temporal decline of chamois body mass is less pronounced in areas with greater proportion of forest cover. For chamois living in forest habitats only, no significant temporal change in body mass was detected. Variation in body mass was affected by the interaction between density and snow cover, as well as by the interaction between spring temperatures and forest cover, supporting the role of forests as thermal buffer against the effects of increasing temperatures on life-history traits in a mountain ungulate. In turn, this study suggests a buffering effect of forests against climate change impacts. Assessments of the consequences of climate change on the life-history traits and population dynamics of mountain-dwelling species should thus consider the plasticity of the species with respect to the use and availability of different habitat types.

Validation of Roebuck 1518 synthetic chamois as a skin simulant when backed by 10% gelatin.

INTRODUCTION: Synthetic skin simulants are used both in wound ballistics and forensic investigations and should display similar mechanical properties to human tissue and therefore need to be validated. It is recognised that skin simulants may have a significantly different performance when different backing combinations are used; therefore, it is essential to specify and control the backing material. Roebuck 1518 synthetic chamois (RBK) backed by 20% ballistic gelatin has been validated as a suitable skin simulant; this study looks at validating the RBK simulant when backed by 10% ballistic gelatin. METHODS: Two layers of RBK synthetic chamois backed by calibrated 10% ballistic gelatin were placed onto the long face of the block and secured. Steel spheres with various sectional densities were fired using a custom-made gas gun to determine the V50 of the simulants and compared with the predicted V50. RESULTS: The results demonstrate that for a sectional density between 2.1 and 6.6 g/cm2, the skin simulants backed by 10% gelatin are within the 35% error bounds predicted by James' patent equation. All samples had a close fit to the regression line (R2 = 0.9738), and a Spearman rho test indicates that there is a "strong" negative correlation between sectional density and the V50 (Rs =- 0.957, p = 0.00). CONCLUSIONS: This validation study confirms that RBK synthetic simulant backed by 10% gelatin is a suitable skin simulant when testing non-deforming projectiles with sectional densities ranging from 2.1 to 6.6 g/cm2. A predictive trend line also indicates that the skin simulant is suitable for non-deforming projectiles with sectional densities ranging from 0.6 to 20 g/cm2 although this needs to be confirmed.

Assessing the role of livestock and sympatric wild ruminants in spreading antimicrobial resistant Campylobacter and Salmonella in alpine ecosystems.

BACKGROUND: Livestock play an important role as reservoir of enteric pathogens and antimicrobial resistance (AMR), a health and economic concern worldwide. However, little is known regarding the transmission and maintenance of these pathogens at the wildlife-livestock interface. In this study, we assessed the occurrence, genetic diversity and AMR of Campylobacter spp. and Salmonella spp. shed by sympatric free-ranging livestock and a wild herbivore in an alpine ecosystem. RESULTS: Campylobacter spp. was isolated from 23.3 % of cattle and 7.7 % of sheep but was not isolated from horses nor Pyrenean chamois (Rupicapra pyrenaica). Campylobacter jejuni was the most frequent species. A high genetic diversity and certain host specificity of C. jejuni isolates was observed. The main AMR detected in Campylobacter isolates was to nalidixic acid (88.2 %), ciprofloxacin (82.4 %) and tetracycline (82.4 %); only 11.7 % of the isolates were pan-susceptible and 17.6 % were multi-resistant. Salmonella ser. Newport was isolated only from one Pyrenean chamois and was pan-susceptible. CONCLUSIONS: Results show that free-ranging cattle and sheep are spreaders of Campylobacter as well as their AMR strains in the alpine environment. Therefore, contaminated alpine pastures or streams may constitute a source for the dissemination of AMR enteropathogens. However, apparently, alpine wild ungulates such as Pyrenean chamois play a negligible role in the epidemiology of zoonotic enteropathogens and AMR, and are not potential bioindicators of the burden of alpine environments.

Microsatellite based assignment reveals history of extirpated mountain ungulate.

During the early 1900s, Northern chamois (Rupicapra rupicapra) populations in the northern Dinaric Mountains were extirpated. During the 1960s and 1970s there were several reintroductions of individuals from two Northern chamois subspecies (Alpine chamois, R. r. rupicapra and Balkan chamois, R. r. balcanica) from neighbouring areas in the attempt to re-establish the population. Accurate taxonomic classification, at subspecies level, of the autochthonous extirpated population was not known. To clarify which subspecies was present before reintroduction, we genotyped four male chamois skulls originating from Velebit Mountain, collected around 25 years before the population local extinction. DNA was successfully extracted from middle layer and outer sheath of horns. Assignment based on microsatellite loci, using both Bayesian clustering in STRUCTURE (with q values between 0.55 and 0.73) and DAPC (with individual membership probabilities of 0.99 and 1.00) indicated higher assessed likelihood for the Alpine subspecies.

Multi-event capture-recapture analysis in Alpine chamois reveals contrasting responses to interspecific competition, within and between populations.

Understanding components of interspecific competition has long been a major goal in ecological studies. Classical models of competition typically consider equal responses of all individuals to the density of competitors, however responses may differ both among individuals from the same population, and between populations. Based on individual long-term monitoring of two chamois populations in sympatry with red deer, we built a multi-event capture-recapture model to assess how vital rates of the smaller chamois are affected by competition from the larger red deer. In both populations, mortality and breeding probabilities of female chamois depend on age and in most cases, breeding status the preceding year. Successful breeders always performed better the next year, indicating that some females are of high quality. In one population where there was high spatial overlap between the two species, the survival of old female chamois that were successful breeders the preceding year (high-quality) was negatively related to an index of red deer population size suggesting that they tend to skip reproduction instead of jeopardizing their own survival when the number of competitors increases. The breeding probability of young breeders (ages 2 and 3) was similarly affected by red deer population size. In contrast, in the second site with low spatial overlap between the two species, the vital rates of female chamois were not related to red deer population size. We provide evidence for population-specific responses to interspecific competition and more generally, for context-, age- and state-dependent effects of interspecific competition. Our results also suggest that the classical assumption of equal responses of all individuals to interspecific competition should be relaxed, and emphasize the need to move towards more mechanistic approaches to better understand how natural populations respond to changes in their environment.

Deer keds on wild ungulates in northern Italy, with a taxonomic key for the identification of Lipoptena spp. of Europe.

Deer keds (Lipoptena spp.) are blood-sucking ectoparasites of domestic and wild animals, and also accidentally of humans. In Europe, five Lipoptena spp. have been recorded, although the lack of specific taxonomic keys has often led to mistaken identification or to missing data. The present study aimed to develop an identification key of the European species and also to identify Lipoptena spp. found on wild ungulates in northern Italy. In total, 390 hippoboscids were collected from Rupicapra rupicapra, Capreolus capreolus, Cervus elaphus and Ovis aries musimon in an Alpine area of Italy. After morphological identification, 140 specimens were subjected to phylogenetic analysis based on mitochondrial (CO1) and nuclear (CAD) gene sequences. Despite the expected presence of slight morphological variations, all specimens examined were identified both microscopically and molecularly as Lipoptena cervi (100% identity for both CO1 and CAD genes). The massive increase in wild ungulate populations can favour the possibility of detecting other species of Lipoptena. The identification keys proposed in the present study may help with monitoring the presence of Lipoptena species, particularly in European countries where this ectoparasite is neglected and for which various data (from diffusion to control methods) are still missing.

Different effects of alpine woody plant expansion on domestic and wild ungulates.

Changes in land-use and climate affect the distribution and diversity of plant and animal species at different spatiotemporal scales. The extent to which species-specific phenotypic plasticity and biotic interactions mediate organismal adaptation to changing environments, however, remains poorly understood. Woody plant expansion is threatening the extent of alpine grasslands worldwide, and evaluating and predicting its effects on herbivores is of crucial importance. Here, we explore the impact of shrubification on the feeding efficiency of Pyrenean chamois (Rupicapra p. pyrenaica), as well as on the three most abundant coexisting domestic ungulate species: cattle, sheep and horses. We use observational diet composition from May to October and model different scenarios of vegetation availability where shrubland and woodland proliferate at the expense of grassland. We then predicted if the four ungulate species could efficiently utilize their food landscapes with their current dietary specificities measuring their niche breath in each scenario. We observed that the wild counterpart, due to a higher trophic plasticity, is less disturbed by shrubification compared to livestock, which rely primarily on herbaceous plants and will be affected 3.6 times more. Our results suggest that mixed feeders, such as chamois, could benefit from fallow landscapes, and that mountain farmers are at a growing economic risk worldwide due to changing land-use practices and climate conditions.

Screening of wild ruminants from the Kaunertal and other alpine regions of Tyrol (Austria) for vector-borne pathogens.

Knowledge about vector-borne pathogens important for human and veterinary medicine in wild ruminants in Tyrol (Austria) is scarce. Blood samples from Alpine ibex (Capra ibex; n = 44), Alpine chamois (Rupicapra rupicapra; n = 21), roe deer (Capreolus capreolus; n = 18) and red deer (Cervus elaphus; n = 6) were collected over a period of 4 years (2015-2018) in four regions in North Tyrol, with a primary focus on the Kaunertal. Blood spots on filter paper were tested for the presence of DNA of vector-borne pathogens (Anaplasmataceae, Piroplasmida, Rickettsia and filarioid helminths). Anaplasma phagocytophilum and Babesia capreoli were detected in two of 89 (2.3%) blood samples. Rickettsia spp., Theileria spp. and filarioid helminths were not documented. One Alpine chamois was positive for A. phagocytophilum and B. capreoli. Moreover, an ibex from the Kaunertal region was positive for A. phagocytophilum. While the ibex was a kid less than 1 year old, the chamois was an adult individual. Further research is recommended to evaluate effects of climate change on infection rates of North Tyrolean wild ruminants by these pathogens and the distribution of their vectors.

A new hybrid approach for MHC genotyping: high-throughput NGS and long read MinION nanopore sequencing, with application to the non-model vertebrate Alpine chamois (Rupicapra rupicapra).

The major histocompatibility complex (MHC) acts as an interface between the immune system and infectious diseases. Accurate characterization and genotyping of the extremely variable MHC loci are challenging especially without a reference sequence. We designed a combination of long-range PCR, Illumina short-reads, and Oxford Nanopore MinION long-reads approaches to capture the genetic variation of the MHC II DRB locus in an Italian population of the Alpine chamois (Rupicapra rupicapra). We utilized long-range PCR to generate a 9 Kb fragment of the DRB locus. Amplicons from six different individuals were fragmented, tagged, and simultaneously sequenced with Illumina MiSeq. One of these amplicons was sequenced with the MinION device, which produced long reads covering the entire amplified fragment. A pipeline that combines short and long reads resolved several short tandem repeats and homopolymers and produced a de novo reference, which was then used to map and genotype the short reads from all individuals. The assembled DRB locus showed a high level of polymorphism and the presence of a recombination breakpoint. Our results suggest that an amplicon-based NGS approach coupled with single-molecule MinION nanopore sequencing can efficiently achieve both the assembly and the genotyping of complex genomic regions in multiple individuals in the absence of a reference sequence.

Low sequence diversity of the prion protein gene (PRNP) in wild deer and goat species from Spain.

The first European cases of chronic wasting disease (CWD) in free-ranging reindeer and wild elk were confirmed in Norway in 2016 highlighting the urgent need to understand transmissible spongiform encephalopathies (TSEs) in the context of European deer species and the many individual populations throughout the European continent. The genetics of the prion protein gene (PRNP) are crucial in determining the relative susceptibility to TSEs. To establish PRNP gene sequence diversity for free-ranging ruminants in the Northeast of Spain, the open reading frame was sequenced in over 350 samples from five species: Iberian red deer (Cervus elaphus hispanicus), roe deer (Capreolus capreolus), fallow deer (Dama dama), Iberian wild goat (Capra pyrenaica hispanica) and Pyrenean chamois (Rupicapra p. pyrenaica). Three single nucleotide polymorphisms (SNPs) were found in red deer: a silent mutation at codon 136, and amino acid changes T98A and Q226E. Pyrenean chamois revealed a silent SNP at codon 38 and an allele with a single octapeptide-repeat deletion. No polymorphisms were found in roe deer, fallow deer and Iberian wild goat. This apparently low variability of the PRNP coding region sequences of four major species in Spain resembles previous findings for wild mammals, but implies that larger surveys will be necessary to find novel, low frequency PRNP gene alleles that may be utilized in CWD risk control.

Impact of climate on the population dynamics of an alpine ungulate: a long-term study of the Tatra chamois Rupicapra rupicapra tatrica.

Global warming is considered as a phenomenon having a negative effect on animals living in cold climate. However, herbivorous species inhabiting cold zones may potentially benefit from increase of temperature as this influence duration of vegetation period and increase food resources. In this study, we analyze the impact of climate factors on the long-term dynamics of an isolated and unhunted population of the Tatra chamois Rupicapra rupicapra tatrica. The population growth rate, based on autumnal chamois counting carried out from 1957 to 2016, were correlated with a set of climatic variables. We tested the hypothesis that high temperatures in summer could have a positive impact on the population, since they influence vegetation growth, which ensure food resources. On the other hand, heavy falls of snow and long-lasting and deep snow cover could adversely affect the population by reducing population survival during the winter. The results of this study indicate that climatic variables best explaining the autoregressed population growth rate (from the autumn of year t-1 to the autumn of year t) were the mean summer temperature of year t-1: the population increase was greater following a warmer summer in year t-1 and, in lower extent, the total precipitation during winter: the population decrease was greater following a winter with heavy snowfall. Duration and thickness of snow cover have no negative effect on population growth rate. The results indicate that the population dynamic of the Tatra chamois is determined in the long term by weather conditions, mainly by temperature, when kids are birthing and growing. The results of this work highlight that climatic changes may be responsible for the population dynamic of high-mountain species. Climate warming may lead to increase in duration of vegetation period in cold climatic zones, what may in turn have positive effect on herbivorous species, which relay upon food resources limited by low temperatures within vegetation period.

Tear production, intraocular pressure and conjunctival bacterial flora in selected captive wild ruminants.

OBJECTIVE: Evaluation of tear production (Schirmer's tear test, STT) and measurement of intraocular pressure (IOP) were performed in a population of captive wild ungulates in a Slovenian ZOO during routine annual health check. ANIMALS STUDIED: In total, 10 fallow deer (Dama dama), 25 mouflons (Ovis aries musimon), 20 alpine ibexes (Capra ibex), and three alpine chamois (Rupicapra rupicapra) were included in the study. METHODS: Tear production was performed by Schirmer's tear test, IOP was measured with an applanation tonometer, and ophthalmological examination was conducted with slit-lamp biomicroscopy and indirect ophthalmoscopy. Conjunctival swabs were taken and submitted for aerobic bacteriology and for detection of Chlamydia spp. and Mycoplasma spp. tested by PCR. RESULTS: Average tear production (in mm/min) was 17.8 ± 3.16 for fallow deer, 17.9 ± 3.87 for mouflons, and 11.7 ± 3.87 for ibexes. Mean intraocular pressure (IOP, in mm Hg) was 14.1 ± 2.48 for fallow deer, 14.9 ± 2.20 for mouflons, and 13.1 ± 2.43 for ibexes. For chamois, average tear production and IOP were 14.5 ± 3.0 and 10.2 ± 2.5, respectively; this is the first record of STT I and IOP in chamois. Bacteriological swabs were positive for bacteria in 100% of the fallow deer, 56% of mouflons, 35% of ibexes, and 100% of chamois. Gram-positive bacteria were predominant. Moraxella spp., Chlamydia spp., and Mycoplasma spp. were not detected. CONCLUSION: The reported values were obtained in animals under manual restraint only to be applicative in similar conditions.

From gestation to weaning: Combining robust design and multi-event models unveils cost of lactation in a large herbivore.

The cost of current reproduction on survival or future reproduction is one of the most studied trade-offs governing resource distribution between fitness components. Results have often been clouded, however, by the existence of individual heterogeneity, with high-quality individuals able to allocate energy to several functions simultaneously, at no apparent cost. Surprisingly, it has also rarely been assessed within a breeding season by breaking down the various reproductive efforts of females from gestation to weaning, even though resource availability and energy requirements vary greatly. We filled this gap by using an intensively monitored population of Pyrenean chamois and by expanding a new methodological approach integrating robust design in a multi-event framework. We distinguished females that gave birth or not, and among reproducing females whether they lost their kid or successfully raised it until weaning. We estimated spring and summer juvenile survival, investigated whether gestation, lactation or weaning incurred costs on the next reproductive occasion, and assessed how individual heterogeneity influenced the detection of such costs. Contrary to expectations if trade-offs occur, we found a positive relationship between gestation and adult survival suggesting that non-breeding females are in poor condition. Costs of reproduction were expressed through negative relationships between lactation and both subsequent breeding probability and spring juvenile survival. Such costs could be detected only once individual heterogeneity (assessed as two groups contrasting good vs. poor breeders) and time variations in juvenile survival were accounted for. Early lactation decreased the probability of future reproduction, providing quantitative evidence of the fitness cost of this period recognized as the most energetically demanding in female mammals and critical for neonatal survival. The new approach employed made it possible to estimate two components of kid survival that are often considered practically unavailable in free-ranging populations, and also revealed that reproductive costs appeared only when contrasting the different stages of reproductive effort. From an evolutionary perspective, our findings stressed the importance of the temporal resolution at which reproductive cost is studied, and also provided insights on the reproductive period during which internal and external factors would be expected to have the greatest fitness impact.

Evidence of circulation of the novel border disease virus genotype 8 in chamois.

Evidence of association between the novel putative border disease virus genotype 8 (BDV-8) and fatal disease in an Alpine chamois (Rupicapra rupicapra) is reported. Diagnostically, we also demonstrated, as already previously reported, the failure of BDV-specific primers (PDB1 and PDB2) to detect BDV-8.