A putatively novel papillomavirus associated with cutaneous plaques and squamous cell carcinoma in captive North American snow leopards (Panthera uncia).

Cutaneous plaques and squamous cell carcinoma (SCC) are common in captive North American snow leopards (SLs) (Panthera uncia). Our objective was to determine whether these lesions are potentially associated with papillomavirus(es). Polymerase chain reaction (PCR) was performed on 3 cutaneous plaques using degenerate primers for papillomaviruses. A putatively novel papillomavirus was identified that shared 76% sequence identity to Felis catus papillomavirus 2. Specific PCR for this virus was performed on 5 cutaneous SCC samples and 7 normal skin samples, which were all positive. In situ hybridization for this putatively novel virus was performed, which revealed strong hybridization signals within hyperplastic cells in cutaneous plaques (n = 3) and within neoplastic cells in cutaneous SCC samples (n = 5). No hybridization signals were identified within normal skin. Ultimately, identification of a causal viral agent in the development of plaques and SCC in SLs will help guide therapeutic intervention and lay the foundation for development of prophylactic vaccines.

Insights into the spatial ecology of severely injured free-living felids: Iberian lynx, bobcat, and snow leopard.

Severe musculoskeletal diseases, such as those associated with congenital or traumatic events, that result in missing limbs may compromise the fitness and survival of free-living felids. Here we report the space use of four amputee individuals from three felid species captured from 2017 to 2022 in Missouri (USA), Toledo and Badajoz (Spain), and Suitai Khairkhan Mountain (Mongolia). We describe home ranges and daily travel distances post-release of free-living felids that had either suffered a traumatic amputation or following a surgical amputation. We compared these data with those reported in the literature for felids without amputations. Forelimb or hindlimb amputation did not affect the hunting, mating, or territory patrolling behavior of any of the individuals. However, we recorded significant differences in the daily movement before and after the traumatic event of the Iberian lynx forelimb amputee. We attribute this difference to the physical impairment, although we consider other variables that may have played a role. Nevertheless, all animals appeared to cope well with their limb loss, showing home ranges and daily distances within those recorded for their sex and species. Unless amputee felids represent a threat to domestic livestock or humans, our data suggest these individuals may remain free-living as they contribute to local population persistence and appear to maintain good general health and welfare.

Spatiotemporal behavior of predators and prey in an arid environment of Central Asia.

The mechanisms of interactions between apex and smaller carnivores may range from competition to facilitation. Conversely, interactions between predators and prey are mainly driven by the prey reducing the likelihood of encounters with predators. In this study, we investigated (1) the spatiotemporal interactions between an apex (the snow leopard) and a meso-predator (the red fox), and (2) the temporal interactions between the snow leopard and its potential prey (Siberian ibex, argali, Asian wild ass, Tolai hare) through camera trapping in the Mongolian Great Gobi-A. The probability of occurrence for the red fox was higher in the presence of the snow leopard than in its absence. Moreover, the red fox activity pattern matched that of the snow leopard, with both species mostly active at sunset. This positive spatiotemporal interaction suggests that the presence of the snow leopard may be beneficial for the red fox in terms of scavenging opportunities. However, other explanations may also be possible. Amongst prey, the Siberian ibex and the argali were mainly active during the day, whereas the Asian wild ass and the Tolai hare were more nocturnal. These findings suggest that potential prey (especially the Siberian ibex and the argali) may shape their behavior to decrease the opportunity for encounters with the snow leopard. Our results have revealed complex interactions between apex and smaller predators and between apex predator and its potential prey.

Retrospective review of eyelid coloboma in snow leopards (Panthera uncia) housed under managed care in North America: 49 cases (2000-2020).

OBJECTIVE: To determine incidence, trends in presentation, associated ocular lesions and other diseases, treatment modalities, and case outcomes of eyelid coloboma cases in snow leopards. ANIMALS: 49 snow leopards with eyelid coloboma living under managed care in North America. METHODS: Medical records were retrospectively searched to identify snow leopards in which eyelid coloboma was diagnosed between January 1, 2000, and December 31, 2020. Data recorded from each animal included signalment, ophthalmic examination information, clinical signs, concurrent health conditions, medical and/or surgical interventions, time to resolution of signs, recurrence of clinical signs, and direct relatives with a history of eyelid coloboma. RESULTS: Ocular clinical signs were present at diagnosis in most cases but not seen in all cases. Corrective procedures were undertaken in 39 cases. Clinical signs were resolved by the total combination of interventions in 84.6% of individuals; however, signs resolved in only 33.3% of cases after a single surgical reconstruction or cryoablation procedure per eye. CLINICAL RELEVANCE: Eyelid coloboma is widespread in the North American snow leopard population. A high percentage (73.5%) have an affected sibling, parent, or grandparent, suggesting a heritable component. Surgical correction resolves or improves clinical signs in most cases, but there is a high rate of postprocedural complications with all procedure types employed. Most complications are minor and manageable, but these can also impact case outcomes. Animals require long-term monitoring, as clinical signs may recur (in some cases, years after initial signs are reported to be resolved), and some animals may require long-term care to manage signs.

Free-ranging livestock affected the spatiotemporal behavior of the endangered snow leopard (Panthera uncia).

Long recognized as a threat to wildlife, particularly for large carnivores, livestock grazing in protected areas can potentially undermine conservation objectives. The interspecific interactions among livestock, snow leopards (Panthera uncia), and their wild prey in fragile Asian highland ecosystems have been a subject of debate. We strategically deployed 164 camera traps in the Wolong National Nature Reserve to systematically investigate the activities of snow leopards, their primary wild ungulate prey species, and free-ranging livestock. We found that snow leopard habitat use was influenced by both wild prey and livestock. Blue sheep served as the main wild prey that spatially attracted snow leopards and coexisted with yaks while free-ranging yaks significantly restricted snow leopard habitat use both temporally and spatially. This study challenges the conventional understanding that livestock indirectly impacts large carnivores by competing with and displacing wild prey. Our findings highlight that free-ranging yaks within the alpine canyon terrain could directly limit snow leopard habitat use, suggesting a potential risk of grazing in reducing apex predator distribution and jeopardizing their populations. Consequently, managing their coexistence in shared habitats requires a more nuanced approach. Furthermore, our research underscores the importance of further research efforts aimed at enhancing our comprehension of the complex interplay within animal communities and ecosystems. This knowledge will contribute to the development of informed, evidence-based conservation strategies and policies.

Genome report: chromosome-level draft assemblies of the snow leopard, African leopard, and tiger (Panthera uncia, Panthera pardus pardus, and Panthera tigris).

The big cats (genus Panthera) represent some of the most popular and charismatic species on the planet. Although some reference genomes are available for this clade, few are at the chromosome level, inhibiting high-resolution genomic studies. We assembled genomes from 3 members of the genus, the tiger (Panthera tigris), the snow leopard (Panthera uncia), and the African leopard (Panthera pardus pardus), at chromosome or near-chromosome level. We used a combination of short- and long-read technologies, as well as proximity ligation data from Hi-C technology, to achieve high continuity and contiguity for each individual. We hope that these genomes will aid in further evolutionary and conservation research of this iconic group of mammals.

Guidelines for Telemetry Studies on Snow Leopards.

Animal-borne tracking devices have generated a wealth of new knowledge, allowing us to better understand, manage and conserve species. Fitting such tracking devices requires that animals are captured and often chemically immobilized. Such procedures cause stress and involve the risk of injuries and loss of life even in healthy individuals. For telemetry studies to be justifiable, it is vital that capture operations are planned and executed in an efficient and ethical way. Project objectives must be clearly articulated to address well-defined knowledge gaps, and studies designed to maximize the probability of achieving those goals. We provide guidelines for how to plan, design, and implement telemetry studies with a special emphasis on snow leopards that are typically captured using foot snares. We also describe the necessary steps to ensure that captures are conducted safely, and with minimal stress to animals.

Detection of SARS-CoV-2 clade B.1.2 in three snow leopards.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is one of seven coronaviruses known to infect humans. Different from other concerned coronavirus and influenza viruses, SARS-CoV-2 has a higher basic reproduction number and thus transmits more efficiently among hosts. Testing animals for SARS-CoV-2 may help decipher virus reservoirs, transmission and pathogenesis. Here, we report the first detection of SARS-CoV-2 in three snow leopards (Panthera uncia) in a zoo in Kentucky in 2020, the first year of the pandemic. Sequence analysis revealed that snow leopard SARS-CoV-2 strains were non-variant B.1.2 lineage and closely correlated with human strains. One snow leopard shed SARS-CoV-2 in faeces up to 4 weeks. Based on clinical signs and viral shedding periods and levels in the three snow leopards, animal-to-animal transmission events could not be excluded. Further testing of SARS-CoV-2 in animals is needed.

On the snow leopard Trails: Occupancy pattern and implications for management in the Pamir.

The snow leopard (Panthera uncia) inhabits one of the most challenging environments on Earth, referred to as the 'third pole'. Only a fraction of its vast range has been explored thus far, owing to myriad of barriers inflicted by the remote terrain and socio-ecological realities of the landscapes. Understanding distribution patterns of species is essential to devise practical management measures. This study aimed to understand the distribution pattern and factors influencing occupancy of snow leopard in the Pamir Mountain range through sign-based occupancy modelling. Our study confirmed that the Pamir range is a snow leopard stronghold, with occupancy estimated at 0.57 ± 0.02. The topographic features positively influenced the detection probability (p = 0.37 ± 0.005) of snow leopards. Occupancy was influenced by mean annual precipitation (ß = -6.12 ± 1.8), density of roads (ß = -1.61 ± 0.6) and water sources (ß = 0.74 ± 0.4). Our findings underpin that sign-based distribution surveys provide vigorous scientific knowledge about elusive species and merit replication being used for other species. We propose to redefine the protected area boundaries based on ecological knowledge and encourage transboundary cooperation to safeguard snow leopards at a landscape scale.

Spatial correlation structures for detections of individuals in spatial capture-recapture models.

Spatial capture-recapture (SCR) models are commonly used to estimate animal density from surveys on which detectors passively detect animals without physical capture, for example, using camera traps, hair snares, or microphones. An individual is more likely to be recorded by detectors close to its activity center, the centroid of its movement throughout the survey. Existing models to account for this spatial heterogeneity in detection probabilities rely on an assumption of independence between detection records at different detectors conditional on the animals' activity centers, which are treated as latent variables. In this paper, we show that this conditional independence assumption may be violated due to the way animals move around the survey region and encounter detectors, such that additional spatial correlation is almost inevitable. We highlight the links between the well-studied issue of unmodeled temporal heterogeneity in nonspatial capture-recapture and this variety of unmodeled spatial heterogeneity in SCR, showing that the latter causes predictable bias in the same way as the former. We address this by introducing a latent detection field into the model, and illustrate the resulting approach with a simulation study and an application to a camera-trap survey of snow leopards Panthera uncia. Our method is a unifying model for several existing SCR approaches, with special cases including standard SCR, models that account for nonspatial individual heterogeneity, and models with overdispersed detection counts.

Increasing risks for emerging infectious diseases within a rapidly changing High Asia.

The cold and arid mountains and plateaus of High Asia, inhabited by a relatively sparse human population, a high density of livestock, and wildlife such as the iconic snow leopard Panthera uncia, are usually considered low risk for disease outbreaks. However, based on current knowledge about drivers of disease emergence, we show that High Asia is rapidly developing conditions that favor increased emergence of infectious diseases and zoonoses. This is because of the existing prevalence of potentially serious pathogens in the system; intensifying environmental degradation; rapid changes in local ecological, socio-ecological, and socio-economic factors; and global risk intensifiers such as climate change and globalization. To better understand and manage the risks posed by diseases to humans, livestock, and wildlife, there is an urgent need for establishing a disease surveillance system and improving human and animal health care. Public health must be integrated with conservation programs, more ecologically sustainable development efforts and long-term disease surveillance.

In the shadows of snow leopards and the Himalayas: density and habitat selection of blue sheep in Manang, Nepal.

There is a growing agreement that conservation needs to be proactive and pay increased attention to common species and to the threats they face. The blue sheep (Pseudois nayaur) plays a key ecological role in sensitive high-altitude ecosystems of Central Asia and is among the main prey species for the globally vulnerable snow leopard (Panthera uncia). As the blue sheep has been increasingly exposed to human pressures, it is vital to estimate its population dynamics, protect the key populations, identify important habitats, and secure a balance between conservation and local livelihoods. We conducted a study in Manang, Annapurna Conservation Area (Nepal), to survey blue sheep on 60 transects in spring (127.9 km) and 61 transects in autumn (134.7 km) of 2019, estimate their minimum densities from total counts, compare these densities with previous estimates, and assess blue sheep habitat selection by the application of generalized additive models (GAMs). Total counts yielded minimum density estimates of 6.0-7.7 and 6.9-7.8 individuals/km2 in spring and autumn, respectively, which are relatively high compared to other areas. Elevation and, to a lesser extent, land cover indicated by the normalized difference vegetation index (NDVI) strongly affected habitat selection by blue sheep, whereas the effects of anthropogenic variables were insignificant. Animals were found mainly in habitats associated with grasslands and shrublands at elevations between 4,200 and 4,700 m. We show that the blue sheep population size in Manang has been largely maintained over the past three decades, indicating the success of the integrated conservation and development efforts in this area. Considering a strong dependence of snow leopards on blue sheep, these findings give hope for the long-term conservation of this big cat in Manang. We suggest that long-term population monitoring and a better understanding of blue sheep-livestock interactions are crucial to maintain healthy populations of blue sheep and, as a consequence, of snow leopards.

Multiple factors influence local perceptions of snow leopards and Himalayan wolves in the central Himalayas, Nepal.

An understanding of local perceptions of carnivores is important for conservation and management planning. In the central Himalayas, Nepal, we interviewed 428 individuals from 85 settlements using a semi-structured questionnaire to quantitatively assess local perceptions and tolerance of snow leopards and wolves. We used generalized linear mixed effect models to assess influential factors, and found that tolerance of snow leopards was much higher than of wolves. Interestingly, having experienced livestock losses had a minor impact on perceptions of the carnivores. Occupation of the respondents had a strong effect on perceptions of snow leopards but not of wolves. Literacy and age had weak impacts on snow leopard perceptions, but the interaction among these terms showed a marked effect, that is, being illiterate had a more marked negative impact among older respondents. Among the various factors affecting perceptions of wolves, numbers of livestock owned and gender were the most important predictors. People with larger livestock herds were more negative towards wolves. In terms of gender, males were more positive to wolves than females, but no such pattern was observed for snow leopards. People's negative perceptions towards wolves were also related to the remoteness of the villages. Factors affecting people's perceptions could not be generalized for the two species, and thus need to be addressed separately. We suggest future conservation projects and programs should prioritize remote settlements.

Meta-replication, sampling bias, and multi-scale model selection: A case study on snow leopard (Panthera uncia) in western China.

Replicated multiple scale species distribution models (SDMs) have become increasingly important to identify the correct variables determining species distribution and their influences on ecological responses. This study explores multi-scale habitat relationships of the snow leopard (Panthera uncia) in two study areas on the Qinghai-Tibetan Plateau of western China. Our primary objectives were to evaluate the degree to which snow leopard habitat relationships, expressed by predictors, scales of response, and magnitude of effects, were consistent across study areas or locally landcape-specific. We coupled univariate scale optimization and the maximum entropy algorithm to produce multivariate SDMs, inferring the relative suitability for the species by ensembling top performing models. We optimized the SDMs based on average omission rate across the top models and ensembles' overlap with a simulated reference model. Comparison of SDMs in the two study areas highlighted landscape-specific responses to limiting factors. These were dependent on the effects of the hydrological network, anthropogenic features, topographic complexity, and the heterogeneity of the landcover patch mosaic. Overall, even accounting for specific local differences, we found general landscape attributes associated with snow leopard ecological requirements, consisting of a positive association with uplands and ridges, aggregated low-contrast landscapes, and large extents of grassy and herbaceous vegetation. As a means to evaluate the performance of two bias correction methods, we explored their effects on three datasets showing a range of bias intensities. The performance of corrections depends on the bias intensity; however, density kernels offered a reliable correction strategy under all circumstances. This study reveals the multi-scale response of snow leopards to environmental attributes and confirms the role of meta-replicated study designs for the identification of spatially varying limiting factors. Furthermore, this study makes important contributions to the ongoing discussion about the best approaches for sampling bias correction.

Assurance of the existence of a trans-boundary population of the snow leopard (Panthera uncia) at Tsagaanshuvuut - Tsagan-Shibetu SPA at the Mongolia-Russia border.

The existence of a trans-boundary population of the snow leopard (Panthera uncia) that inhabits the massifs of Tsagaanshuvuut (Mongolia) - Tsagan-Shibetu (Russia) was determined through non-invasive genetic analysis of scat samples and by studying the structure of territory use by a collared female individual. The genetic analysis included species identification of samples through sequencing of a fragment of the cytochrome b gene and individual identification using a panel of 8 microsatellites. The home range of a female snow leopard marked with a satellite Global Positioning System (GPS) collar was represented by the minimum convex polygon method (MCP) 100, the MCP 95 method and the fixed kernel 95 method. The results revealed insignificant genetic differentiation between snow leopards that inhabit both massifs (minimal fixation index [FST ]), and the data testify to the unity of the cross-border group. Moreover, 5 common individuals were identified from Mongolian and Russian territories. This finding clearly shows that their home range includes territories of both countries. In addition, regular movement of a collared snow leopard in Mongolia and Russia confirmed the existence of a cross-border snow leopard group. These data support that trans-boundary conservation is important for snow leopards in both countries. We conclude that it is crucial for Russia to study the northern range of snow leopards in Asia.

A pilot study-genetic diversity and population structure of snow leopards of Gilgit-Baltistan, Pakistan, using molecular techniques.

BACKGROUND: The Hindu Kush and Karakoram mountain ranges in Pakistan's northern areas are a natural habitat of the snow leopard (Panthera uncia syn. Uncia uncia) but the ecological studies on this animal are scarce since it is human shy by nature and lives in difficult mountainous tracts. The pilot study is conducted to exploit the genetic diversity and population structure of the snow leopard in this selected natural habitat of the member of the wildcat family in Pakistan. METHOD: About 50 putative scat samples of snow leopard from five localities of Gilgit-Baltistan (Pakistan) along with a control sample of zoo maintained male snow leopard were collected for comparison. Significant quality and quantity of genomic DNA was extracted from scat samples using combined Zhang-phenol-chloroform method and successful amplification of cytochrome c oxidase I gene (190 bp) using mini-barcode primers, seven simple sequence repeats (SSR) markers and Y-linked AMELY gene (200 bp) was done. RESULTS: Cytochrome c oxidase I gene sequencing suggested that 33/50 (66%) scat samples were of snow leopard. AMELY primer suggested that out of 33 amplified samples, 21 (63.63%) scats were from male and 12 (36.36%) from female leopards. Through successful amplification of DNA of 25 out of 33 (75.75%) scat samples using SSR markers, a total of 68 alleles on seven SSR loci were identified, showing low heterozygosity, while high gene flow between population. DISCUSSION: The low gene flow rate among the population results in low genetic diversity causing decreased diversification. This affects the adaptability to climatic changes, thus ultimately resulting in decreased population size of the species.

LIP-TO-LID TRANSPOSITION AND HUSBANDRY MANAGEMENT FOR SEVERE BILATERAL EYELID COLOBOMA IN THREE SNOW LEOPARDS (PANTHERA UNCIA).

Three sibling snow leopard (Panthera uncia) cubs were evaluated shortly after birth and found to have bilateral eyelid colobomas involving >50% of the upper central and lateral lid margins. The cubs also had iris to cornea persistent pupillary membranes and developed varying degrees of keratitis caused by trichiasis and exposure. No fundic abnormalities were noted. Given the severity of the defects an extensive surgical correction was required. Prior to surgical correction, the cubs were separated from the dam daily and a human-animal socialization plan was enacted to prepare the cubs for the intensive postoperative care that would be required. Bilateral lip commissure to eyelid transposition surgeries were performed on one cub at a time at 2 wk intervals starting at 3 mo of age. Postoperative care was labor and time intensive and each cub was maintained at the zoo's veterinary hospital for 11-21 days with daily supervised visits with the cub's siblings. All cubs were successfully reintroduced together and with the dam at the end of each hospitalization. The surgical procedure yielded fully functional eyelids and an excellent cosmetic appearance in all three cubs. The planned consistent human interaction with the cubs before and during the postoperative period made the procedure and its intensive aftercare possible and was crucial to the successful outcome of the surgeries.

OSTEOCHONDRAL AUTOGRAFT TRANSFER FOR TREATMENT OF STIFLE OSTEOCHONDRITIS DISSECANS IN TWO RELATED SNOW LEOPARDS ( PANTHERA UNCIA).

Two related female snow leopards ( Panthera uncia) were diagnosed with osteochondritis dissecans of the lateral femoral condyle via radiographs and computed tomography. Lesions were either bilateral (Patient 1) or unilateral (Patient 2). Surgical treatment was performed using an osteochondral autograft transfer system following cartilage flap removal. After a 6-wk recovery period, both animals had a markedly improved gait with mild pelvic limb stiffness. Patient 1 developed intermittent lameness 9 mo postsurgery and Patient 2 had mild lameness 3 yr postsurgery. Because of recurrent lameness, both animals were treated with platelet-rich plasma intra-articular injections and oral gabapentin (Patient 1) or oral meloxicam (Patient 2). Full incorporation of autografts was confirmed with computed tomography (both patients) and arthroscopy (Patient 1 only). Osteochondritis dissecans has been previously reported in snow leopards and these cases represent the first reported use of osteochondral autograft repair and platelet-rich plasma to treat the condition.

Range-Wide Snow Leopard Phylogeography Supports Three Subspecies.

The snow leopard, Panthera uncia, is an elusive high-altitude specialist that inhabits vast, inaccessible habitat across Asia. We conducted the first range-wide genetic assessment of snow leopards based on noninvasive scat surveys. Thirty-three microsatellites were genotyped and a total of 683 bp of mitochondrial DNA sequenced in 70 individuals. Snow leopards exhibited low genetic diversity at microsatellites (AN = 5.8, HO = 0.433, HE = 0.568), virtually no mtDNA variation, and underwent a bottleneck in the Holocene (∼8000 years ago) coinciding with increased temperatures, precipitation, and upward treeline shift in the Tibetan Plateau. Multiple analyses supported 3 primary genetic clusters: (1) Northern (the Altai region), (2) Central (core Himalaya and Tibetan Plateau), and (3) Western (Tian Shan, Pamir, trans-Himalaya regions). Accordingly, we recognize 3 subspecies, Panthera uncia irbis (Northern group), Panthera uncia uncia (Western group), and Panthera uncia uncioides (Central group) based upon genetic distinctness, low levels of admixture, unambiguous population assignment, and geographic separation. The patterns of variation were consistent with desert-basin "barrier effects" of the Gobi isolating the northern subspecies (Mongolia), and the trans-Himalaya dividing the central (Qinghai, Tibet, Bhutan, and Nepal) and western subspecies (India, Pakistan, Tajikistan, and Kyrgyzstan). Hierarchical Bayesian clustering analysis revealed additional subdivision into a minimum of 6 proposed management units: western Mongolia, southern Mongolia, Tian Shan, Pamir-Himalaya, Tibet-Himalaya, and Qinghai, with spatial autocorrelation suggesting potential connectivity by dispersing individuals up to ∼400 km. We provide a foundation for global conservation of snow leopard subspecies, and set the stage for in-depth landscape genetics and genomic studies.