Abstract: This project is part of an ongoing snow leopard study established in 2003 with support from the ISLT. The study involves a multifaceted approach designed to provide important baseline data on the status of the snow leopard population in Langtang National Park (LNP), Nepal and to generate long-term support and commitment to the conservation of snow leopards in the park. The specific aims include: 1) conducting a population survey of the snow leopards in LNP, focusing on distribution and abundance; 2) assessing the status of prey species populations in the park; and 3) providing educational outreach programs on snow leopard conservation for local school children (K-8) living in the park. During the 2004 study period, snow leopard signs were observed (including pugmarks and scats) although somewhat fewer than in 2003. Similarly, the average herd size of the snow leopards' primary prey species in LNP (the Himalayan thar) was a bit lower than in 2003. There is speculation that the thar populations and the snow leopards may be moving to more remotes areas of the park perhaps in response to increasing pressure from domestic livestock grazing. This possibility is being addressed during the 2005 study period.

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.

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.

Abstract: Livestock production is the major land-use in Ladakh region of the Indian Trans-Himalaya, and is a crucial sector that drives the region's economy (Anon, 2002). Animal products like meat and milk provide protein to the diet of people, while products like wool and pashmina (soft fibre of goats) find their way to the international market. Such high utility of livestock and the recent socio-economic changes in the region have caused an increase in livestock population (Rawat and Adhikari, 2002; Anon. 2002), which, if continue apace, may increase grazing pressure and deteriorate pasture conditions. Thus, there is an urgent need to assess the impact of such escalation in livestock population on the regions wildlife. Although, competitive interaction between wildlife and livestock has been studied elsewhere in the Trans-Himalaya (Bhatnagar et al., 2000; Mishra, 2001; Bagchi et al., 2002), knowledge on this aspect in the Ladakh region is very rudimentary. The rangelands of Ladakh are characterised by low primary productivity (Chundawat & Rawat, 1994), and the wild herbivores are likely to compete with the burgeoning livestock on these impoverished rangelands (Mishra et al., 2002). Thus, given that the area supports a diverse wild ungulate assemblage of eight species (Fox et al., 1991b), and an increasing livestock population (Rawat and Adhikari, 2002), the nature of interaction between wildlife and livestock needs to be assessed. During this project, we primarily evaluated the influence of domestic sheep and goat grazing on the habitat use of Tibetan argali Ovis ammon hodgsoni in a prospective wildlife reserve in Ladakh.

Abstract: The potential for rescuing immature oocytes from the ovaries of females of rare felid species which die or undergo medical ovariohysterectomy was evaluated. Ovaries were recovered from 13 species representing 35 individuals in good-to-poor health. Although the majority of females were 10 yr of age or older and in fair-to-poor health, a total of 846 oocytes were recovered of which 608 (71.9%) were classified as fair-to- excellent quality. One hundred of these oocytes were used for initial maturation classification and as parthogenetic controls. Overall, of the 508 fair-to-excellent quality oocytes placed in culture, 164 (32.3%) matured to metaphase II in vitro. For species in which 3 or more individuals yielded oocytes, mean oocyte maturation rates were as follows: 36.2%, tiger; 27.9% leopard; and 8.3%, cheetah. In vitro insemination of oocytes resulted in fertilization (2 polar bodies, 2 pronuclei, or cleavage) rates of 9.1% to 28.6% (leopard) using homologous fresh spermatozoa and 4.0% (lion) to 40.0% (puma) using homologous frozen-thawed spermatozoa. Inseminations using heterologous (domestic cat) spermatozoa also resulted in fertilized oocytes in the tiger, leopard, snow leopard, puma, serval, and Geoffroy's cat (range in fertilization rate, 5.0% for leopard to 46.2% for puma). Cleaved embryos resulted from the insemination of leopard oocytes with homologous sperm (n = 1 embryo) and puma oocytes with domestic cat sperm (n = 3 embryos). These results demonstrate that immature ovarian oocytes from rare felid species can be stimulated to mature in vitro despite an excision-to-culture interval as long as 36 h.(ABSTRACT TRUNCATED AT 250 WORDS)