Rode, J., Pelletier, A., Fumey, J., Rode, S., Cabanat, A. L., Ouvrard, A., Chaix, B., White, B., Harnden, M., Xuan, N. T., Vereshagin, A., Casane, D. (2020). Diachronic monitoring of snow leopards at Sarychat-Ertash State Reserve (Kyrgyzstan) through scat genotyping: a pilot study. bioRxiv, , 1–21.
Abstract: Snow leopards (Panthera uncia) are a keystone species of Central Asia’s high mountain ecosystem. The species is listed as vulnerable and is elusive, preventing accurate population assessments that could inform conservation actions. Non-invasive genetic monitoring conducted by citizen scientists offers avenues to provide key data on this species that would otherwise be inaccessible. From 2011 to 2015, OSI-Panthera citizen science expeditions tracked signs of presence of snow leopards along transects in the main valleys and crests of the Sarychat-Ertash State Reserve (Kyrgyzstan). Scat samples were genotyped at seven autosomal microsatellite loci and at a X/Y locus for sex identification, which allowed estimating a minimum of 11 individuals present in the reserve from 2011 to 2015. The genetic recapture of 7 of these individuals enabled diachronic monitoring, providing indications of individuals’ movements throughout the reserve. We found putative family relationships between several individuals. Our results demonstrate the potential of this citizen science program to get a precise description of a snow leopard population through time.
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Gosselin, S. J., Loudy, D. L., Tarr, M. J., Balistreri, W. F., Setchell, K. D., Johnston, J. O., et al. (1988). Veno-occlusive disease of the liver in captive cheetah. Vet Pathol, 25(1), 48–57.
Abstract: Liver tissues from 126 captive cheetah were evaluated by light microscopy and histochemistry; eight animals were evaluated by electron microscopy. The main hepatic lesion, a vascular lesion resembling veno- occlusive disease (VOD) of the liver and characterized by subendothelial fibrosis and proliferation of smooth muscle-like cells in the central veins, was seen in 60% of the sexually mature cheetah. Although this hepatic vascular lesion was seen in cheetah as young as 1 year of age, the most severe lesions, usually associated with liver failure, were found in cheetah between the ages of 6 and 11. There was no sex predisposition, and in approximately 40% of the VOD cases, liver disease was not suspected clinically or at necropsy. VOD was found in other felidae, especially in the snow leopard. High levels of vitamin A in livers, as well as in diets of the cheetah, could be a contributing factor in the development of VOD in some groups of cheetah.
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De Groot, H., Van Swieten, P., & Aalberse, R. C. (1990). Evidence for a Fel d I-like molecule in the “big cats” (Felidae species). J Allergy Clin Immunol, 86(1), 107–116.
Abstract: In this study, we investigated the cross-reactivity pattern of IgE and IgG4 antibodies to the major feline allergen, Fel d I. We studied the IgE and IgG4 response of 11 cat-allergic patients against Fel d I-like structures in eight members of the Felidae family: ocelot, puma, serval, siberian tiger, lion, jaguar, snow leopard, and caracal. Hair from these “big cats” was collected, extracted, and used in a RAST system and histamine-release test. By means of a RAST-inhibition assay with affinity-purified Fel d I from cat dander, it was established that, in the Felidae species, a Fel d I equivalent is present that reacts with IgE and IgG4 antibodies. We found that all patients had cross-reacting IgE antibodies to seven of the Felidae tested; no IgE antibodies reactive with the caracal were found. Eight of 10 patients with IgG4 antibodies directed to cat dander also had IgG4 antibodies directed to several Felidae species, including the caracal. However, the correlation between the IgE and the IgG4 antibody specificity was low, indicating that, in the case of Fel d I IgE and IgG4, antibodies do not necessarily have the same specificity.
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Brown, J. L., Wasser, S. K., Wildt, D. E., & Graham, L. H. (1994). Comparative Aspects of Steroid Hormone Metabolism and Ovarian Activity in Felids, Measured Noninvasively in Feces. Biol Reprod, 51(4), 776–786.
Abstract: Noninvasive fecal assays were used to study steroid metabolism and ovarian activity in several felid species. Using the domestic cat (Felis catus) as model, the excretory products of injected [14C]estradiol (E2) and [14C]progesterone (P4) were determined. Within 2 days, 97.0 +/- 0.6% and 96.7 +/- 0.5% of recovered E2 and P4 radioactivity, respectively, was found in feces. E2 was excreted as unconjugated estradiol and estrone (40%) and as a non-enzyme- hydrolyzable conjugate (60%). P4 was excreted primarily as non-enzyme- hydrolyzable, conjugated metabolites (78%) and as unconjugated pregnenolone epimers. A simple method for extracting fecal steroid metabolites optimized extraction efficiencies of the E2 and P4 excretion products (90.1 +/- 0.8% and 87.2 +/- 1.4%, respectively). Analysis of HPLC fractions of extracted fecal samples from the radiolabel-injected domestic cats revealed that E2 immunoreactivity coincided primarily with the unconjugated metabolized [14C]E2 peak, whereas progestogen immunoreactivity coincided with a single conjugated epimer and multiple unconjugated pregnenolone epimers. After HPLC separation, similar immunoreactive E2 and P4 metabolite profiles were observed in the leopard cat (F. bengalensis), cheetah (Acinonyx jubatus), clouded leopard (Neofelis nebulosa), and snow leopard (Panthera uncia). Longitudinal analyses demonstrated that changes in fecal E2 and P4 metabolite concentrations reflected natural or artificially induced ovarian activity. For example, severalfold increases in E2 excretion were associated with overt estrus or exogenous gonadotropin treatment, and elevated fecal P4 metabolite concentrations occurred during pregnant and nonpregnant (pseudopregnant) luteal phases. Although overall concentrations were similar, the duration of elevated fecal P4 metabolites during pseudopregnancy was approximately half that observed during pregnancy. In summary, steroid metabolism mechanisms appear to be conserved among these physically diverse, taxonomically related species. Results indicate that this hormone-monitoring approach will be extremely useful for elucidating the hormonal regulatory mechanism associated with the reproductive cycle, pregnancy, and parturition of intractable and endangered felid species.
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Korablev, M. P., Poyarkov, A. D., Karnaukhov, A. S., Zvychaynaya, E. Y., Kuksin, A. N., Malykh, S. V., Istomov, S. V., Spitsyn, S. V., Aleksandrov, D. Y., Hernandez-Blanco, J. A., Munkhtsog, B., Munkhtogtokh, O., Putintsev, N. I., Vereshchagin, A. S., Becmurody, A., Afzunov, S., Rozhnov, V. V. (2021). Large-scale and fine-grain population structure and genetic diversity of snow leopards (Panthera uncia Schreber, 1776) from the northern and western parts of the range with an emphasis on the Russian population. Conservation Genetics, .
Abstract: The snow leopard (Panthera uncia Schreber, 1776) population in Russia and Mongolia is situated at the northern edge of the range, where instability of ecological conditions and of prey availability may serve as prerequisites for demographic instability and, consequently, for reducing the genetic diversity. Moreover, this northern area of the species distribution is connected with the western and central parts by only a few small fragments of potential habitats in the Tian-Shan spurs in China and Kazakhstan. Given this structure of the range, the restriction of gene flow between the northern and other regions of snow leopard distribution can be expected. Under these conditions, data on population genetics would be extremely important for assessment of genetic diversity, population structure and gene flow both at regional and large-scale level. To investigate large-scale and fine-grain population structure and levels of genetic diversity we analyzed 108 snow leopards identified from noninvasively collected scat samples from Russia and Mongolia (the northern part of the range) as well as from Kyrgyzstan and Tajikistan (the western part of the range) using panel of eight polymorphic microsatellites. We found low to moderate levels of genetic diversity in the studied populations. Among local habitats, the highest heterozygosity and allelic richness were recorded in Kyrgyzstan (He = 0.66 ± 0.03, Ho = 0.70 ± 0.04, Ar = 3.17) whereas the lowest diversity was found in a periphery subpopulation in Buryatia Republic of Russia (He = 0.41 ± 0.12, Ho = 0.29 ± 0.05, Ar = 2.33). In general, snow leopards from the western range exhibit greater genetic diversity (He = 0.68 ± 0.04, Ho = 0.66 ± 0.03, Ar = 4.95) compared to those from the northern range (He = 0.60 ± 0.06, Ho = 0.49 ± 0.02, Ar = 4.45). In addition, we have identified signs of fragmentation in the northern habitat, which have led to significant genetic divergence between subpopulations in Russia. Multiple analyses of genetic structure support considerable genetic differentiation between the northern and western range parts, which may testify to subspecies subdivision of snow leopards from these regions. The observed patterns of genetic structure are evidence for delineation of several management units within the studied populations, requiring individual approaches for conservation initiatives, particularly related to translocation events. The causes for the revealed patterns of genetic structure and levels of genetic diversity are discussed.
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Chetri, M., Odden, M., Sharma, K., Flagstad, O., Wegge, P. (2019). Estimating snow leopard density using fecal DNA in a large landscape in north-central Nepal. Global Ecology and Conservation, (17), 1–8.
Abstract: Although abundance estimates have a strong bearing on the conservation status of a
species, less than 2% of the global snow leopard distribution range has been sampled
systematically, mostly in small survey areas. In order to estimate snow leopard density
across a large landscape, we collected 347 putative snow leopard scats from 246 transects
(490 km) in twenty-six 5 5km sized sampling grid cells within 4393 km2 in Annapurna-
Manaslu, Nepal. From 182 confirmed snow leopard scats, 81 were identified as belonging
to 34 individuals; the remaining were discarded for their low (<0.625) quality index. Using
maximum likelihood based spatial capture recapture analysis, we developed candidate
model sets to test effects of various covariates on density and detection of scats on transects.
The best models described the variation in density as a quadratic function of
elevation and detection as a linear function of topography. The average density estimate of
snow leopards for the area of interest within Nepal was 0.95 (SE 0.19) animals per 100 km2
(0.66e1.41 95% CL) with predicted densities varying between 0.1 and 1.9 in different parts,
thus highlighting the heterogeneity in densities as a function of habitat types. Our density
estimate was low compared to previous estimates from smaller study areas. Probably,
estimates from some of these areas were inflated due to locally high abundances in overlap
zones (hotspots) of neighboring individuals, whose territories probably range far beyond
study area borders. Our results highlight the need for a large-scale approach in snow
leopard monitoring, and we recommend that methodological problems related to spatial
scale are taken into account in future snow leopard research.
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Janecka, J. E., Jackson, R., Munkhtsog, B., Murphy, W. J. (2014). Characterization of 9 microsatellites and primers in snow leopards and a species-specific PCR assay for identifying noninvasive samples. Conservation Genetic Resource, 6(2), 369:373.
Abstract: Molecular markers that can effectively identify noninvasively collected samples and provide genetic
information are critical for understanding the distribution, status, and ecology of snow leopards (Panthera uncia). However, the low DNA quantity and quality in many
noninvasive samples such as scats makes PCR amplification and genotyping challenging. We therefore designed primers for 9 microsatellites loci previously isolated in the
domestic cat (Felis catus) specifically for snow leopard studies using noninvasive samples. The loci showed moderate levels of variation in two Mongolian snow leopard
populations. Combined with seven other loci that we previously described, they have sufficient variation (He = 0.504, An = 3.6) for individual identification and
population structure analysis. We designed a species species specific PCR assay using cytochrome b for identification of unknown snow leopard samples. These molecular markers
facilitate in depth studies to assess distribution, abundance, population structure, and landscape connectivity of this endangered species.
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Janecka, J. E., Jackson, R., Munkhtsog, B., Murphy, W. J. (2014). Characterization of 9 microsatellites and primers in snow leopards and a species-specific PCR assay for identifying noninvasive samples. Conservation Genetic Resource, 6(2), 369:373.
Abstract: Molecular markers that can effectively identify noninvasively collected samples and provide genetic
information are critical for understanding the distribution, status, and ecology of snow leopards (Panthera uncia). However, the low DNA quantity and quality in many
noninvasive samples such as scats makes PCR amplification and genotyping challenging. We therefore designed primers for 9 microsatellites loci previously isolated in the
domestic cat (Felis catus) specifically for snow leopard studies using noninvasive samples. The loci showed moderate levels of variation in two Mongolian snow leopard
populations. Combined with seven other loci that we previously described, they have sufficient variation (He = 0.504, An = 3.6) for individual identification and
population structure analysis. We designed a species species specific PCR assay using cytochrome b for identification of unknown snow leopard samples. These molecular markers
facilitate in depth studies to assess distribution, abundance, population structure, and landscape connectivity of this endangered species.
endangered species
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Rodgers, T. W., Janecka, J. E. (2013). Applications and techniques for non-invasive faecal genetics research in felid conservation.
Abstract: Non-invasive genetic techniques utilising DNA extracted from faeces hold great promise for felid conservation research. These methods can be used to establish species
distributions, model habitat requirements, analyse diet, estimate abundance and population density, and form the basis for population, landscape and conservation genetic analyses. Due to the elusive nature of most felid species, non-invasive genetic methods have the potential to provide
valuable data that cannot be obtained with traditional observational or capture techniques. Thus, these methods are particularly valuable for research and conservation of endangered
felid species. Here, we review recent studies that use non-invasive faecal genetic techniques to survey or study wild felids; provide an overview of field, laboratory and analysis techniques; and offer suggestions on how future non-invasive genetic studies can be expanded or improved to more effectively support conservation.
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Lama, R. P., Ghale, T. R., Suwal, M. K., Ranabhat, R., Regmi, G. R. (2018). First photographic evidence of Snow Leopard Panthera uncia (Mammalia: Carnivora: Felidae) outside current protected areas network in Nepal Himalaya. Journal of Threatened Taxa, , 12086–12090.
Abstract: The Snow Leopard Panthera uncia is a rare top predator of high-altitude ecosystems and insufficiently surveyed outside of protected areas in Nepal. We conducted a rapid camera-trapping survey to assess the presence of Snow Leopard in the Limi valley of Humla District. Three individuals were recorded in two camera locations offering the first photographic evidence of this elusive cat outside the protected area network of Nepal. In addition to Snow Leopard, the Blue Sheep Pseudois nayaur, Beech Marten Martes foina, Pika Ochotona spp. and different species of birds were also detected by camera-traps. More extensive surveys and monitoring are needed for reliably estimating the population size of Snow Leopard in the area. The most urgent needs are community-based conservation activities aimed at mitigating immediate threats of poaching, retaliatory killing, and rapid prey depletion to ensure the survival of this top predator in the Himalaya.
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