Mitochondrial cytochrome b indicates the presence of two paraphyletic diverged lineages of the blue sheep Pseudois nayaur across the Indian Himalaya: conservation implications.

BACKGROUND: Populations exhibit signatures of local adaptive traits due to spatial and environmental heterogeneity resulting in microevolution. The blue sheep is widely distributed across the high Asian mountains and are the snow leopard's principal prey species. These mountains differ in their evolutionary history due to differential glaciation and deglaciation periods, orography, and rainfall patterns, and such factors causes diversification in species. METHODS AND RESULTS: Therefore, we assess the phylogeographic status of blue sheep using the mitochondrial cytochrome b gene (220 bp) across the Indian Himalayan region (IHR) and its relationship with other populations. Of the observed five haplotypes, two and three were from the western Himalayas (WH) and eastern Himalayas (EH) respectively. One of the haplotypes from WH was shared with the population of Pamir plateau, suggesting historical maternal connectivity between these areas. The phylogenetic analyses split the blue sheep into two paraphyletic clades, and western and eastern populations of IHR were within the Pamir and Tibetan plateau clades, respectively. We observed a relatively higher mean sequence divergence in the EH population than in the WH. CONCLUSION: We propose five 'Evolutionary Significant Units' across the blue sheep distribution range based on observed variation in the species' ecological requirements, orography, climatic conditions, and maternal lineages, viz.; Western Himalaya-Pamir plateau (WHPP); Eastern Himalaya-Tibetan plateau (EHTP); Qilian mountains; Helan mountains and Hengduan mountains population. Despite the small sample size, population divergence was observed across the IHR, therefore, we suggest a transboundary, collaborative study on comparative morphology, anatomy, ecology, behaviour, and population genetics using harmonized different genetic markers for identifying the overall taxonomic status of the blue sheep across its range for planning effective conservation strategies.

Comparison of fecal microbiota composition of blue sheep fed Lolium perenne versus Sorghum sudanense.

Lolium perenne L. and Sorghum sudanense (Piper) Stapf. are 2 common forages fed to blue sheep (Pseudois nayaur Hodgson, 1833) in captivity. However, the effect of these 2 forages on the gastrointestinal microbiota is largely unknown. We analyzed the diversity of the microbiota in the feces of captive blue sheep fed with L. perenne (group F1) and S. sudanense (group F2) by 16S rRNA sequencing. A total of 20 major phyla and 29 genera fecal bacterial communities were detected in the 2 groups. The F1 and F2 groups shared common microbiota at the phylum level, which mainly consisted of Firmicutes and Bacteroidetes. Ruminococcaceae_UCG-005, Eubacterium_coprostanoligenes_group, Ruminococcaceae_UCG-013, and Ruminococcaceae_UCG-010 were the top 4 dominant taxa at the genus level. The percentage of Ruminococcaceae_UCG-010 was significantly higher in the F2 group (∼2.75-fold) than in F1 group. The diversity and abundance of the microbial community in F2 was higher than that in F1. Although both of L. perenne and S. sudanense affect the metabolism of the gastrointestinal microbiota of blue sheep, the S. sudanense improves more aspects of metabolism and biogenesis. In summary, our results demonstrated that L. perenne and S. sudanense affect blue sheep gastrointestinal microbiota in different ways. But S. sudanense efficiently improved the gastrointestinal microbiota of blue sheep.

First isolation of Trichuris from wild blue sheep (Pseudois nayaur) in the Helan Mountains, China.

Four adult female worms of Trichuris were isolated from an individual of the wild blue sheep (Pseudois nayaur) inhabiting the Helan Mountains, China, during an epidemiological survey of this wild ruminant. Although there were some differences among the worms in posterior end (rectum) morphology and egg shape, little information regarding species status could be inferred from their morphology. Phylogenetic trees were constructed based on sequences of the ITS1 segment of ribosomal RNA (rRNA), and the sequences of the four Trichuris specimens from wild blue sheep were divided into two distinct lineages (Clade A and Clade B). The two specimens in Clade A were named Genotype I, and had the closest relationship with Trichuris skrjabini; the two specimens in Clade B were named Genotype II and had the closest genetic relationship with a previously described Trichuris sp. In the two Trichuris genotypes identified in the present study, the 18S fragments (261 to 262 bp) of the newly obtained sequences were found to be highly conserved, with merely one insert mutation of a single nucleotide present. The genetic distance based on ITS1 between members of Clade A, composed of two T. skrjabini individuals and two Genotype I individuals, ranged from 0 to 0.0034. These distances are within the intraspecies variation of Trichuris (0-0.0272), suggesting that the Genotype I individuals infesting the wild blue sheep were T. skrjabini. In Clade B, the newly obtained sequences clustered with Trichuris sp. specimens isolated from ruminants (sheep and black goat) with strong support, and the genetic distance ranged from 0.0068 to 0.017, which is also within the intraspecies variation of Trichuris (0-0.0272). However, the genetic distances between the Clade A and Clade B were 0.0442 to 0.0578, which are higher than the intraspecies distances in Trichuris but lower than the interspecies distances (0.102-0.5078). These results implied that Clade A and Clade B most likely represent two subpopulations of T. skrjabini; however, the possibility that Clade A is T. skrjabini and Clade B is a Candidatus Trichuris could not be excluded.

A detection of benzimidazole resistance-associated SNPs in the isotype-1 ß-tubulin gene in Haemonchus contortus from wild blue sheep (Pseudois nayaur) sympatric with sheep in Helan Mountains, China.

BACKGROUND: Benzimidazole (BZ) resistance is an increasingly serious problem due to the excessive use of this anthelmintic for controlling Haemonchus contortus, which is one of the major gastrointestinal nematodes infecting small ruminants worldwide. Three known single nucleotide polymorphisms (SNPs), F167Y (TAC), E198A (GCA) and F200Y (TAC), in the isotype-1 ß-tubulin gene of H. contortus are associated with BZ resistance. Comprehending the spread and origins of BZ resistance-associated SNPs has important implications for the control of this nematode. RESULTS: Twenty-seven adult H. contortus were harvested from wild blue sheep (Pseudois nayaur), small wild ruminants sympatric with domestic ruminants, inhabiting the Helan Mountains, China, to monitor the status of BZ resistance. In addition, 20 adult H. contortus from domestic sheep sympatric with this wild ruminant and 36 isotype-1 ß-tubulin haplotype sequences of H. contortus (two of these haplotypes, E198A3 and E198A4, possessed resistance-associated SNP E198A (GCA) from domestic ruminants in eight other geographical regions of China were used to further define the origins of BZ resistance-associated SNPs within the worms collected from blue sheep. The BZ resistance-associated SNP E198A was detected, whereas SNPs F167Y (TAC) and F200Y (TAC) were not found within the worms collected from blue sheep, and the frequency of homozygous resistant E198A (GCA) was 7.40%. The evolutionary tree and network showed consistent topologies for which there was no obvious boundary among the worms from the wild and domestic hosts, and two haplotypes (E198A1 and E198A2) possessing E198A from the wild blue sheep had two different independent origins. E198A1 had the same origin with E198A3 but E198A2 had a different origin with them. Population genetic analyses revealed a low level of Fst values (ranging from 0 to 0.19749) between all H. contortus worm groups in China. CONCLUSIONS: Results of the current study of the three BZ resistance-associated SNPs of H. contortus from wild blue sheep suggested that only E198A (GCA) was present within the worms collected from the wild ruminants and had multiple independent origins.

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.

Growth Stages and Inter-Species Gut Microbiota Composition and Function in Captive Red Deer (Cervus elaphus alxaicus) and Blue Sheep (Pseudois nayaur).

Blue sheep and red deer, second-class key protected animals in China, are sympatric species with a high degree of overlap of food resources in the Helan Mountains, China. Previous studies with blue sheep and red deer in nature have shown that their physiology is closely related to their gut microbiota. However, growth stages and changes occurring in these species in captivity are still unknown. Thus, 16S rRNA gene sequencing was used to explore diversity, composition and function of the gut microbiota in these two animal species. The diversity and structure of the gut microbiota in captive blue sheep and red deer changed at different growth stages, but the dominant microbiota phyla in the gut microbiota remained stable, which was composed of the phyla Firmicutes, Bacteroidetes and Verrucomicrobia. Moreover, gut microbiota diversity in juvenile blue sheep and red deer was low, with the potential for further colonization. Functional predictions showed differences such as red deer transcription being enriched in adults, and blue sheep adults having a higher cell wall/membrane/envelope biogenesis than juveniles. Microbial changes between blue sheep and red deer at different growth stages and between species mainly depend on the abundance of the microbiota, rather than the increase and absence of the bacterial taxa.

No silver bullet? Snow leopard prey selection in Mt. Kangchenjunga, Nepal.

In this study, we investigated the impact of domestic and wild prey availability on snow leopard prey preference in the Kangchenjunga Conservation Area of eastern Nepal-a region where small domestic livestock are absent and small wild ungulate prey are present. We took a comprehensive approach that combined fecal genetic sampling, macro- and microscopic analyses of snow leopard diets, and direct observation of blue sheep and livestock in the KCA. Out of the collected 88 putative snow leopard scat samples from 140 transects (290 km) in 27 (4 × 4 km2) sampling grid cells, 73 (83%) were confirmed to be from snow leopard. The genetic analysis accounted for 19 individual snow leopards (10 males and 9 females), with a mean population size estimate of 24 (95% CI: 19-29) and an average density of 3.9 snow leopards/100 km2 within 609 km2. The total available prey biomass of blue sheep and yak was estimated at 355,236 kg (505 kg yak/km2 and 78 kg blue sheep/km2). From the available prey biomass, we estimated snow leopards consumed 7% annually, which comprised wild prey (49%), domestic livestock (45%), and 6% unidentified items. The estimated 47,736 kg blue sheep biomass gives a snow leopard-to-blue sheep ratio of 1:59 on a weight basis. The high preference of snow leopard to domestic livestock appears to be influenced by a much smaller available biomass of wild prey than in other regions of Nepal (e.g., 78 kg/km2 in the KCA compared with a range of 200-300 kg/km2 in other regions of Nepal). Along with livestock insurance scheme improvement, there needs to be a focus on improved livestock guarding, predator-proof corrals as well as engaging and educating local people to be citizen scientists on the importance of snow leopard conservation, involving them in long-term monitoring programs and promotion of ecotourism.

The Discontinuous Elevational Distribution of an Ungulate at the Regional Scale: Implications for Speciation and Conservation.

The elevational range where montane species live is a key factor of spatial niche partitioning, because the limits of such ranges are influenced by interspecies interaction, abiotic stress, and dispersal barriers. At the regional scale, unimodal distributions of single species along the elevation gradient have often been reported, while discontinuous patterns, such as bimodal distributions, and potential ecological implications have been rarely discussed. Here, we used extensive camera trap records to reveal the elevation distribution of Himalaya blue sheep (Pseudois nayaur) and its co-existence with other ground animal communities along a slope of Baima Snow Mountain, southwest China. The results show that Himalaya blue sheep exhibited a distinctive bimodal distribution along the elevation gradient contrasting the unimodal distributions found for the other ungulates in Baima snow mountain. A first distributional peak was represented by a population habituating in scree habitat around 4100 m, and a second peak was found in the dry-hot valley around 2600 m. The two distinct populations co-existed with disparate animal communities and these assemblages were similar both in the dry and rainy seasons. The extremely low abundance of blue sheep observed in the densely forested belt at mid-elevation indicates that vegetation rather than temperature is responsible for such segregation. The low-elevation population relied highly on Opuntia ficus-indica, an invasive cactus species that colonized the region six hundred years ago, as food resource. Being the only animal that developed a strategy to feed on this spiky plant, we suggest invasive species may have formed new foraging niche to support blue sheep population in lower elevation hot-dry river valleys, resulting in the geographic separation from the original population and a potential morphological differentiation, as recorded. These findings emphasize the important conservation values of role of ecological functions to identify different taxa, and conservation values of apparent similar species of different ecological functions.

Hoof Pressure Distribution Pattern of Blue Sheep During Walking on Different Slopes: A Subject-Specific Analysis.

The purpose of this study was to quantitatively assess the vertical force distribution (VFD) of subject-specific healthy blue sheep while walking on different slopes using a pressure-sensing walkway. The blue sheep was trained to walk over the pressure-sensing walkway by choosing a comfortable walking speed, and the slope angle increased from 0° to 25°. The sheep's hooves were divided into four quadrants, namely, the cranio-lateral, cranio-medial, caudo-lateral, and caudo-medial quadrants, to investigate the VFD of the peak vertical force (PVF), vertical impulse (VI) and occurrence time of the PVF during the stance phase (TPVF). This study demonstrates that the main stressed quadrant of the front hoof changes from the caudo-medial quadrant to the cranio-medial quadrant with increasing slope. The main stressed quadrant of the rear hoof is the cranio-medial quadrant and does not change with the increasing slope. For all the slopes, the vertical force shifted from the lateral quadrant to the medial quadrant and from the caudal quadrant to the cranial quadrant. All the results obtained in the study suggest the feasibility of detecting gait changes in blue sheep, which has potential for the diagnosis of lower limb musculoskeletal diseases in quadrupeds.

The Passive Contact Stability of Blue Sheep Hoof Based on Structure, Mechanical Properties, and Surface Morphology.

As the only component that contacts the ground and rock, the hooves of blue sheep may play a crucial role in their excellent climbing abilities. In this study, we used a combination of techniques, including scanning electron microscopy, infrared spectroscopy and nanoindentation, to characterize the surface morphology, structure, material composition, and mechanical properties of blue sheep hoof and investigate the potential contributions of these properties to the establishment of passive contact stability. Straight and curled microscopic lamellar morphology were found on the hoof surfaces. The cross section of the hoof revealed four layers, and each layer had a unique structure. Finite element analysis was employed to verify that the surface morphology and microstructure effectively contributed to the slip resistance and impact cushioning, respectively. Analyses of the energy and infrared spectra showed that the organic and inorganic substances in different regions of the hoof had similar components but different contents of those components. The hoof was mainly composed of keratin. From the outside to the inside, gradients in both the modulus and hardness were observed. These factors help the hoof alleviate high impact strengths and increase contact stability. These findings further our understanding of the unique mechanism of blue sheep hoof and may help in the development of novel biomimetic materials and mechanical components with enhanced friction and contact stability properties.

Taxonomic status of Chinese blue sheep (Pseudois nayaur): new evidence of a distinct subspecies.

The blue sheep is an endemic species to the Qinghai-Tibet Plateau and surrounding regions. It has been regarded as having 2 subspecies: Pseudois nayaur nayaur and P. n. szechuanensis. However, such a classification remains controversial. Herein, we analyze 10 microsatellite loci and part of the mitochondrial control region for clarification in such taxonomic debates. We use samples from 168 individuals from 6 geographic populations covering almost all the distribution areas of the species in China to carry out comparisons. Phylogenetic trees derived from both the microsatellite and mitochondrial markers combined with the discriminant analysis of principal components (DAPC) and the STRUCTURE analysis reveal that the individuals in the Helan Mountains are well grouped with a distinct evolutionary lineage and are significantly different from the other populations of P. n. szechuanensis according to Fst values, implying that this isolated population should be categorized as a valid subspecies; namely, Pseudois nayaur alashanicus. The isolation-by-distance (IBD) analysis shows a significant positive relationship between genetic and geographical distances among the populations.

Seasonal Variation and Sexual Dimorphism of the Microbiota in Wild Blue Sheep (Pseudois nayaur).

Microbiota of the wild blue sheep (Pseudois nayaur) presents a seasonal variation due to different dietary selection and feeding strategies from different ecological niches chosen by different sex in summer. To address those issues, we analyzed the variation of gut microbiota based on the material from the feces, with 16S rRNA and meta-genome aimed to explore seasonal and gender differences. The results indicate that seasonal dietary changes and gender differentiation, as expected, cause the variation in sheep's gut microbiota structure. The variation of the former is more significant than the latter. Dominant Firmicutes exists a significantly higher abundance in summer than that in winter. Subordinate Bacteroides expresses no seasonal difference between the two seasons. Compared with the winter group, the summer group is featured by abundant enzymes digesting cellulose and generating short-chain fatty acids (SCFAs), such as beta-glucosidase (EC: 3.2.1.21) for cellulose digestion, and butyrate kinase (EC:2.7.2.7) in butyrate metabolism, implying that the changes of the composition in intestinal flora allow the sheep to adapt to the seasonalized dietary selection through alternated microbial functions to reach the goal of facilitating the efficiency of energy harvesting. The results also show that the blue sheep expresses a prominent sexual dimorphism in the components of gut microbiota, indicating that the two sexes have different adaptations to the dietary selection, and demands for physical and psychological purposes. Thus, this study provides an example of demonstrating the principles and regulations of natural selection and environmental adaptation.

Comparative Analyses of Fecal Microbiota in European Mouflon (Ovis orientalis musimon) and Blue Sheep (Pseudois nayaur) Living at Low or High Altitudes.

The gut microbiota is a complex and essential system organ that plays an integrative role in balancing key vital functions in the host. Knowledge of the impact of altitude on the gut microbiota of European mouflon (Ovis orientalis musimon) and blue sheep (Pseudois nayaur) is currently limited. In this study, we compared the characteristics of gut microbiota in 5 mouflon at low altitude (K group), 4 mouflon at high altitude (L group), 4 blue sheep at low altitude (M group), and 4 blue sheep at high altitude (N group). The V3-V4 region of the 16S rRNA gene was analyzed using high-throughput sequencing. Analyses based on the operational taxonomic units showed significant changes in the gut microbial communities between groups at different altitudes. At the phylum level, groups at the high altitudes had a higher relative abundance of Firmicutes and a lower relative abundance of Bacteroidetes than those at the low altitudes. A higher Firmicutes:Bacteroidetes ratio is beneficial to animals in terms of the gut microbiota-mediated energy harvest. The relative abundance of Proteobacteria was significantly higher in the gut microbiota of mouflon sheep at high altitudes. At the genus level, the Bacteroides:Prevotella ratio was significantly higher in the low-altitude group (than the high-altitude group) of mouflon sheep and the ratio was significantly higher in the high-altitude group (than the low-altitude group) in blue sheep. In addition, the Ruminococcaceae_UCG-005 related to cellulose and starch digestion was the predominant genus in blue sheep and the relative abundance of the genus was significant higher in the high-altitude group than the low-altitude group of blue sheep (P < 0.01). In conclusion, our results suggested that the gut microbiota of high-altitude groups of sheep had stronger abilities related to energy metabolism and the decomposition of substances, e.g., fiber and cellulose, and that such abilities are associated with high-altitude adaptation.

Molecular phylogeny and phylogeography of genus Pseudois (Bovidae, Cetartiodactyla): New insights into the contrasting phylogeographic structure.

Blue sheep, Pseudois nayaur, is endemic to the Tibetan Plateau and the surrounding mountains, which are the highest-elevation areas in the world. Classical morphological taxonomy suggests that there are two subspecies in genus Pseudois (Bovidae, Artiodactyla), namely Pseudois nayaur nayaur and Pseudois nayaur szechuanensis. However, the validity and geographic characteristics of these subspecies have never been carefully discussed and analyzed. This may be partially because previous studies have mainly focused on the vague taxonomic status of Pseudois schaeferi (dwarf blue sheep). Thus, there is an urgent need to investigate the evolutionary relationship and taxonomy system of this genus. This study enriches a previous dataset by providing a large number of new samples, based on a total of 225 samples covering almost the entire distribution of blue sheep. Molecular data from cytochrome b and the mitochondrial control region sequences were used to reconstruct the phylogeny of this species. The phylogenetic inferences show that vicariance plays an important role in diversification within this genus. In terms of molecular dating results and biogeographic analyses, the striking biogeographic pattern coincides significantly with major geophysical events. Although the results raise doubt about the present recognized distribution range of blue sheep, they have corroborated the validity of the identified subspecies in genus Pseudois. Meanwhile, these results demonstrate that the two geographically distinct populations, the Helan Mountains and Pamir Plateau populations, have been significantly differentiated from the identified subspecies, a finding that challenges the conventional taxonomy of blue sheep.

Genetic diversity of Haemonchus contortus isolated from sympatric wild blue sheep (Pseudois nayaur) and sheep in Helan Mountains, China.

BACKGROUND: Haemonchus contortus is known among parasitic nematodes as one of the major veterinary pathogens of small ruminants and results in great economic losses worldwide. Human activities, such as the sympatric grazing of wild with domestic animals, may place susceptible wildlife hosts at risk of increased prevalence and infection intensity with this common small ruminant parasite. Studies on phylogenetic factors of H. contortus should assist in defining the amount of the impact of anthropogenic factors on the extent of sharing of agents such as this nematode between domestic animals and wildlife. METHODS: H. contortus specimens (n = 57) were isolated from wild blue sheep (Pseudois nayaur) inhabiting Helan Mountains (HM), China and additional H. contortus specimens (n = 20) were isolated from domestic sheep that were grazed near the natural habitat of the blue sheep. Complete ITS2 (second internal transcribed spacer) sequences and partial sequences of the nad4 (nicotinamide dehydrogenase subunit 4 gene) gene were amplified to determine the sequence variations and population genetic diversities between these two populations. Also, 142 nad4 haplotype sequences of H. contortus from seven other geographical regions of China were retrieved from database to further examine the H. contortus population structure. RESULTS: Sequence analysis revealed 10 genotypes (ITS2) and 73 haplotypes (nad4) among the 77 specimens, with nucleotide diversities of 0.007 and 0.021, respectively, similar to previous studies in other countries, such as Pakistan, Malaysia and Yemen. Phylogenetic analyses (BI, MP, NJ) of nad4 sequences showed that there were no noticeable boundaries among H. contortus populations from different geographical origin and population genetic analyses revealed that most of the variation (94.21%) occurred within H. contortus populations. All phylogenetic analyses indicated that there was little genetic differentiation but a high degree of gene flow among the H. contortus populations among wild blue sheep and domestic ruminants in China. CONCLUSIONS: The current work is the first genetic characterization of H. contortus isolated from wild blue sheep in the Helan Mountains region. The results revealed a low genetic differentiation and high degree of gene flow between the H. contortus populations from sympatric wild blue sheep and domestic sheep, indicating regular cross-infection between the sympatrically reared ruminants.

The complete mitochondrial genome sequence of the dwarf blue sheep, Pseudois schaeferi haltenorth in China.

The dwarf blue sheep (Pseudois schaeferi haltenorth) belongs the subfamily Caprinae, which is distributed in Sichuan, Tibet, Yunnan, and Qinghai in China. In this study, the complete mitochondrial genome of Pseudois schaeferi haltenorth was sequenced. The mitogenome was 16 741 bp in length, consisting of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and a non-coding control region (D-loop region). As in other mammals, most mitochondrial genes are encoded on the heavy strand, except for ND6 and eight tRNA genes which are encoded on the light strand. The overall base composition of the Pseudois schaeferi haltenorth is 33.54% A, 26.37% T, 26.91% C, and 13.18% G, A + T (59.91%) was higher than G + C (40.09%). The phylogenetic relationships was analyzed using the complete mitogenome sequence, results show that P. schaeferi haltenorth should be a different species differ from the Genus pseudois hodgson. These information provide useful data for further study on the protection of genetic resources and the taxonomy of Caprinae.

Blue sheep in the Annapurna Conservation Area, Nepal: habitat use, population biomass and their contribution to the carrying capacity of snow leopards.

The Himalaya region of Nepal provides a habitat for the endangered snow leopard (Panthera uncia) and its principal prey species, the blue sheep (Pseudois nayaur). The aim of this study was to describe the habitat, the distribution and the population structure of blue sheep, and to estimate their contribution to the carrying capacity of snow leopard in the upper Mustang region of Nepal. Blue sheep were recorded at altitudes from 3209-5498 m on slopes with gradients of 16-60° and aspects of 40°NE to 140°SE. A total of 939 blue sheep were counted in the upper Mustang region, and 98 were counted in the Yak Kharka region of Manang district; however, upper Mustang had the lowest population density of blue sheep recorded within their distribution range in Nepal (0.86 blue sheep/km(2)). The results of the study show that a higher density of blue sheep is associated with greater plant species diversity. The most important species present in the blue sheep habitat were Kobresia pygmaea, Artemesia spp., Lonicera spp., Lancea tibetica, Poa spp., Astragalus spp. and Ephedra gerardiana. It is estimated that the existing blue sheep population biomass of approximately 38 925 kg in the upper Mustang region could support approximately 19 snow leopards (1.6 snow leopards/100 km(2)).