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.

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: Twenty-five years ago, the snow leopard Uncia uncia, an endangered large cat, was eliminated from what is now Sagarmatha National Park (SNP). Heavy hunting pressure depleted that area of most medium-large mammals, before it became a park. After three decades of protection, the cessation of hunting and the recovery of wild ungulate populations, snow leopards have recently returned (four individuals). We have documented the effects of the return of the snow leopard on the population of its main wild prey, the Himalayan tahr Hemitragus jemlahicus, a 'near-threatened' caprin. Signs of snow leopard presence were recorded and scats were collected along a fixed trail (130 km) to assess the presence and food habits of the snow leopard in the Park, from 2004 to 2006. Himalayan tahr, the staple of the diet, had a relative occurrence of 48% in summer and 37% in autumn, compared with the next most frequent prey, musk deer Moschus chrysogaster (summer: 20%; autumn: 15%) and cattle (summer: 15%; autumn: 27%). In early summer, the birth rate of tahr (young-to-female ratio: 0.8-0.9) was high. The decrease of this ratio to 0.1-0.2 in autumn implied that summer predation concentrated on young tahr, eventually altering the population by removing the kid cohort. Small populations of wild Caprinae, for example the Himalayan tahr population in SNP, are sensitive to stochastic predation events and may be led to almost local extinction. If predation on livestock keeps growing, together with the decrease of Himalayan tahr, retaliatory killing of snow leopards by local people may be expected, and the snow leopard could again be at risk of local extinction. Restoration of biodiversity through the return of a large predator has to be monitored carefully, especially in areas affected by humans, where the lack of important environmental components, for example key prey species, may make the return of a predator a challenging event.

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.