Population survey and genetic diversity of snow leopards Panthera uncia as revealed by fecal DNA

Snow leopards Panthera uncia occur in rugged,high altitude regions of Central Asia,where they are endangered as a result of human induced factors including low prey densities and poaching.Information on the status of this felid is limited in many regions.We examined the feasibility of using noninvasive genetic methods to monitor snow leopard populations and to study their genetic diversity.We observed snow leopard scats in three separate geographic regions during brief surveys and collected 109 scats from southwestern India(Ladakh),western China(Qinghai),and southern Mongolia(South Gobi).We used a panel of mitochondrial DNA(mtDNA)primers and seven domestic cat(Felis catus)microsatellite loci to survey the snow leopard population and to examine their genetic diversity.Of the 32 scat samples found in Ladakh,50 found in Qinghai,and 27 found in Mongolia,80% produced positive species identification.Positive snow leopard identification was made in 17 of 32 scats collected in Ladakh,where 6 red fox and 2 wolf/dog scats were also detected.In Qinghai,36 scats were identified by mtDNA analysis,including 3 snow leopard,10 lynx,21 red fox,and 2 wolf/dog.Only snow leopard(11)and red fox(13)scats were observed in South Gobi.We genotyped 31 confirmed snow leopard scats with the 7 microsatellites selected.There were a total of 9 unique genotypes detected:4 in Ladakh and 5 in South Gobi.Using the Y-linked AMELY marker,we identified 2 males and 2 females in Ladakh,and 3 males and 2 females in South Gobi.All 7 loci were polymorphic in both Ladakh and South Gobi with numbers of alleles ranging from 2 to 4.No loci were found to be out of HW equilibrium after sequential Bonferroni correction of significance levels.The numbers of alleles and heterozygosity were lower in South Gobi,although our sample sizes were too small to test for significance.The F_ST estimated between the two areas was 0.171.Our findings highlight the efficacy of noninvasive genetic surveys for monitoring snow leopards and the feasibility of large-scale studies that will provide critical information for conservation of snow leopards throughout Central Asia.