Abstract: 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 dif!cult 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 !ve localities of Gilgit-Baltistan (Pakistan) along with a control sample of zoo maintained male snow leopard were collected for comparison. Signi!cant quality and quantity of genomic DNA was extracted from scat samples using combined Zhang�phenol�chloroform method and successful ampli!cation 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 ampli!ed samples, 21 (63.63%) scats were from male and 12 (36.36%) from female leopards. Through successful ampli!cation of DNA of 25 out of 33 (75.75%) scat samples using SSR markers, a total of 68 alleles on seven SSR loci were identi!ed, showing low heterozygosity, while high gene "ow between population.
Discussion: The low gene flow rate among the population results in low genetic diversity causing decreased diversi!cation. This affects the adaptability to climatic changes, thus ultimately resulting in decreased population size of the species.

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: Snow leopards Panthera uncia are under threat across their range and require urgent conservation actions based on sound science. However, their remote habitat and cryptic nature make them inherently difficult to study and past attempts have provided insufficient information upon which to base effective conservation. Further, there has been no statistically-reliable and cost-effective method available to monitor snow leopard populations, focus conservation effort on key populations, or assess conservation impacts. To address these multiple information needs, Panthera, Snow Leopard Trust, and Snow Leopard Conservation Fund, launched an ambitious long-term study in Mongolia’s South Gobi province in 2008. To date, 10 snow leo-pards have been fitted with GPS-satellite collars to provide information on basic snow leopard ecology. Using 2,443 locations we calculated MCP home ranges of 150 – 938 km2, with substantial overlap between individuals. Exploratory movements outside typical snow leopard habitat have been observed. Trials of camera trapping, fecal genetics, and occupancy modeling, have been completed. Each method ex-hibits promise, and limitations, as potential monitoring tools for this elusive species.

Abstract: Most conventional spermatology research involves common mammalian species including livestock, laboratory animals and humans. Yet, there are more than 4500 mammalian species inhabiting the planet for which little is known about basic reproductive biology, including sperm characteristics and function. This information is important, not just as adjunct knowledge, but because the majority of these species are threatened with extinction, largely due to human-induced pressures. The field of conservation is changing rapidly, and global cooperation is emerging among a variety of wildlife enthusiasts, ranging from management authorities of nature reserves to curators of rare zoological collections. Conservation progress depends on systematic, multidisciplinary research first to answer basic questions, with new data then applied to endangered species management plans. The reproductive physiologist is a crucial component of this scheme. Reproduction is the essence of species survival, and enormous effort needs to be directed at these 'untraditional' research species, subspecies and populations. Spermatology research combined with simultaneous efforts in endocrinology, embryology and cryopreservation (among others) can lead to the successful application of assisted reproduction. Examples from this laboratory include an array of wild felid species and a rare cervid and mustelid. Obstacles to success are formidable, including unique species-specificities, diminished genetic diversity and a general lack of resources. Nonetheless, the field offers tremendous opportunities for generating unique knowledge of comparative interest and with conservation utility.

Abstract: An extraction and assay procedure to measure fecal estradiol-17-beta and progestin concentrations in several cat species was developed and validated for use for noninvasive monitoring of ovarian function. Fecal samples were collected over a range of 3-20 months from female tigers (three), lions (three), snow leopards (three), cheetahs (two), caracals (two), and domestic cats (five). Samples were extracted with 90% methanol, lipids removed with petroleum ether, and the estradiol and progestins in the methanol measured by radioimmunoassay (RIA). High Performance Liquid Chromatography (HPLC) fractionation and subsequent RIA of the fractions indicated that the estradiol-17-beta antiserum cross-reacted primarily with estradiol-17-beta in the feces of lions and tigers and was assumed to be specific for estradiol-17-beta in the feces of other species as well. However, there were several immunoreactive compounds, presumably progesterone metabolites, excreted in the feces which varied both quantitatively and qualitatively among species. The behavior of tigers, lions, cheetahs, and caracals was visually monitored during the collection period and frequency of sexual behaviors was positively correlated with increases in fecal estradiol in all species observed. The mean fecal estradiol-17-beta peaks were as follows: tigers, 128.0 +- 13.1; lions, 186.0 +- 14.8; snow leopards, 136.7 +- 15.9; cheetahs, 140.9 +- 9.0; caracals, 24.5 +- 4.0; and domestic cats 158.9 +- 19.3 ng/gm. Fecal progestin concentrations rose significantly (P lt 0,001) only after breeding or during pregnancy and were as follows: tigers, 5.6 +- 0.6; lions, 1.9 +- 0.1; cheetahs, 8.4 +- 1.1; and caracals, 2.4 +- 0.4 mu-g/gm. Fecal progestins were elevated for one-half to two-thirds of the gestation length during presumed pseudopregnancy but remained elevated throughout successful pregnancies. These results suggest that ovarian function can be monitored noninvasively in the family Felidae by the measurement of fecal estradiol-17-beta and progestin concentrations.