SUBSPECIES IDENTIFICATION, GENETIC VARIATION, AND CONSERVATION OF THE CLOUDED LEOPARD (Neofelis nebulosa)

Abstract

The clouded leopard, Neofelis nebulosa, is one of the largest felids in Southeast Asia and is currently threatened by human persecution, habitat loss and fragmentation. We used modern molecular genetic techniques to examine the validity of traditionally used taxonomic nomenclature and investigated the molecular genetic variation for the clouded leopard in order to gain insight on its evolutionary history. We examined 51 microsatellites, and 3,900 base pairs of mitochondria' and nuclear genes to assess and compare genetic variation in 108 clouded leopards, 118 individuals of the five Panthera genus species, and 3 domestic cats. Clouded leopards showed similar levels of microsatellite genetic variation to the individual species of Panthera, but low levels of mtDNA and nuclear variation relative to species within Panthera genus. Evidence of divergences was found between the subspecies N. n. diardi and the mainland subspecies in all the nuclear and mitochondrial analyses. These differences were supported by a karyotype analysis where comparisons showed a distinct difference in the F3 chromosome between N. n. nebulosa and N. n. diardi. Based upon the molecular data and karyotype analysis, we recommend that the Bomean clouded leopards be classified as a distinct species. Conservation strategies for clouded leopards and the Bornean species should be reevaluated to incorporate the new taxonomic units.