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Buckley-Beason, V.A.; Johnson, W.E.; Nash, W.G.; Stanyon, R.; Menninger, J.C.; Driscoll, C.A.; Howard, J.G.; Bush, M.; Page, J.E.; Roelke, M.; Stone, G.; Martelli, P.P.; Wen, C.; Ling, L.; Duraisingam, R.K.; Lam, P.V.; O'Brien, S.J.
Molecular evidence for species-level distinctions in clouded leopards
2006  Current Biology (16): 2371-2376

Among the 37 living species of Felidae, the clouded leopard (_Neofelis nebulosa_) is generally classified as a monotypic genus basal to the Panthera lineage of great cats [1-5]. This secretive, mid-sized (16-23 kg) carnivore, now severely endangered, is traditionally subdivided into four southeast Asian subspecies (Figure 1A) [4-8].We used molecular genetic methods to re-evaluate subspecies partitions and to quantify patterns of population genetic variation among 109 clouded leopards of known geographic origin (Figure 1A, Tables S1 and S2 in the Supplemental Data available online). We found strong phylogeographicmonophyly and large genetic distances between N. n. nebulosa (mainland) and N. n. diardi (Borneo; n = 3 individuals) with mtDNA (771 bp), nuclear DNA (3100 bp), and 51 microsatellite loci. Thirty-six fixed mitochondrial and nuclear nucleotide differences and 20 microsatellite loci with nonoverlapping allele-size ranges distinguished N. n. nebulosa from _N. n. diardi_. Along with fixed subspecies-specific chromosomal differences, this degree of differentiation is equivalent to, or greater than, comparable measures among fiverecognized Panthera species (lion, tiger, leopard, jaguar, and snow leopard). These distinctions increase the urgency of clouded leopard conservation efforts, and if affirmed by morphological analysis and wider sampling of_ N. n. diardi_ in Borneo and Sumatra, would support reclassification of _N. n. diardi_ as a new species (_Neofelis diardi_).

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