Abstract: The endangered snow leopard Panthera uncia occurs in rugged, high-altitude regions of Central Asia. However, information on the status of this felid is limited in many areas. We conducted a pilot study to optimize molecular markers for the analysis of snow leopard scat samples and to examine the feasibility of using noninvasive genetic methods for monitoring this felid. We designed snow leopard-specific primers for seven microsatellite loci that amplified shorter segments and avoided flanking sequences shared with repetitive elements. By redesigning primers we maximized genotyping success and minimized genotyping errors. In addition, we tested a Y chromosome-marker for sex identification and designed a panel of mitochondrial DNA primers for examining genetic diversity of snow leopards using scat samples. We collected scats believed to be from snow leopards in three separate geographic regions including north-western India, central China and southern Mongolia. We observed snow leopard scats in all three sites despite only brief 2-day surveys in each area. There was a high rate of species misidentification in the field with up to 54% of snow leopard scats misidentified as red fox. The high rate of field misidentification suggests sign surveys incorporating scat likely overestimate snow leopard abundance. The highest ratio of snow leopard scats was observed in Ladakh (India) and South Gobi (Mongolia), where four and five snow leopards were detected, respectively. Our findings describe a species-specific molecular panel for analysis of snow leopard scats, and highlight the efficacy of noninvasive genetic surveys for monitoring snow leopards. These methods enable large-scale noninvasive studies that will provide information critical for conservation of snow leopards.

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.

Abstract: The endangered snow leopard (Panthera uncia) is widely but sparsely distributed throughout the mountainous regions of central Asia. Detailed information on the status and abundance of the snow leopard is limited because of the logistical challenges faced when working in the rugged terrain it occupies, along with its secretive nature. Camera-trapping and noninvasive genetic techniques have been used successfully to survey this felid. We compared noninvasive genetic and camera-trapping snow leopard surveys in the Gobi Desert of Mongolia. We collected 180 putative snow leopard scats from 3 sites during an 8-day period along 37.74 km of transects. We then conducted a 65-day photographic survey at 1 of these sites, approximately 2 months after scat collection. In the site where both techniques were used noninvasive genetics detected 5 individuals in only 2 days of fieldwork compared to 7 individuals observed in the 65-day camera-trapping session. Estimates of population size from noninvasive genetics ranged between 16 and 19 snow leopards in the 314.3-km2 area surveyed, yielding densities of 4.9–5.9 individuals/100 km2. In comparison, the population estimate from the 65-day photographic survey was 4 individuals (adults only) within the 264-km2 area, for a density estimate of 1.5 snow leopards/100 km2. Higher density estimates from the noninvasive genetic survey were due partly to an inability to determine age and exclude subadults, reduced spatial distribution of sampling points as a consequence of collecting scats along linear transects, and deposition of scats by multiple snow leopards on common sites. Resulting differences could inflate abundance estimated from noninvasive genetic surveys and prevent direct comparison of densities derived from the 2 approaches unless appropriate adjustments are made to the study design.

Abstract: The recent development of potent and specifica lphar-adrenoceptoar gonistsa nd antagonists has enhanced their use in nondomestic animal immobilization and reversal. Medetomidine, a new potent alphar-agonist, in combination with the dissociative anesthetic ketamine, has been used to immobilize a variety of nondomestic mammals. Medetomidine alone induces sedation in a dose-dependent way, and complete immobilization has been achieved with high doses in semidomesticated reindeer (Rangifer tarandus) and blue foxes (Alopex lagopus). Howbver, we feel that ketamine should be added to the immobilization mixture to ensure complete immobilization and operator safety. In ketamine combinations, medetomidine doses are usually 60-100 pg/kg. The required ketamine doses are remarkably low:0.8-1.6 mglkg in most ruminants,2.5-3.0 mgUgin felids,u rsids,a nd canids,a nd 5.G-8.0m glkgi n primates,w olverines(Gulog ulo),ando therm uitelids. Clinically, the resulting immobilization is characterized by a smooth onset, good to excellent myorelaxation, and areflexia at higher doses. Determinations of hematologic, serum biochemicil, arterial blood gas,a nd acid-bases tatusp arametersi ndicate that the immobilization is physiologically sound. We have had no fatalities attributable to the immobilization mixture ( I ,240 immobilizations). The alphar-adrenoceptora ntagonist,a tipamezole,i s highly efective in reversingt he immobilization induced by medetomidine, medetomidine-ketamine combinations, or xylazine. In ruminants, the medetomidine-ketamine-induced immobilization can be rapidly and persistently reversed by administering 100-l 50 1rg/kg of alipamezole i.v. and the rest s.c., adjusting the total atipamezole dose to an atipamezole: medetomidine ratio of approximately 4-5 (w/w). Becauseth e required ketamine doses are relatively high in carnivores, we prefer to use a lower atipamezole dose (totil atipamezoie: medetomidine ratio approximately 2-3 w/w) and to administer it i.m. or s.c. Using thii regimen, reversals are calm and animals show minimal “residual ketamine effect.” Because atipamezole is a competitive antagonist, its dose should be reduced if it is administered late in the immobilization period when a large part of medetomidine has been endogenously metabolized. Xylazine-induced immobilization is rapidly reversed by I mg of atipamezole for every 8-12 mg of xylazine used.