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Buckley-Beason, V.A.; Johnson, W.E.; Nash, W.G.; Stanyon, R.; Menninger, J.C.; Driscoll, C.A.; Howard, J.; Bush, M.; Page, J.E.; Roelke, M.E.; Stone, G.; Martelli, P.P.; Wen, C.; Ling, L.; Duraisingam, R.K.; Lam, P.V.; O'Brien, S.J.
Molecular Evidence for Species-Level Distinction in Modern Clouded Leopards (_Neofelis nebulosa_)
2007  Conference Proceeding

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. This secretive, midsized (16-23 kg) carnivore, now severely endangered, is traditionally subdivided into four southeast Asian subspecies. 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. We found strong phylogeographic monophyly and large genetic distances between _N. n. nebulosa _(mainland) and _N. n. diardi_ (Borneo; n=3) with mtDNA, nuclear DNA, 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 five recognized 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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