Isolation of

Natural infection with Leishmania spp. in phlebotomine sandflies was searched for during a longitudinal study carried out from July 1997 to July 1998 in the village Catarnica, Municipality Independencia, Táchira State. This hamlet is an old endemic focus of cutaneous leishmaniasis in the Venezuelan Andean region, which lies close to the Colombian border at 1,300 m a.s.l., in an agricultural area mainly used for cultivating coffee. Phlebotomine sandflies were collected using Shannon traps placed in the peridomestic habitat from 19:00 to 21:00 hs. Males were stored in alcohol 70% while females were kept in Nunc vials with 10% DMSO and cryopreserved in liquid nitrogen for subsequent dissection and identification. The most abundant anthropophilic species was Lutzomyia spinicrassa with 3,032 males and 4,290 females (85.4%). Among 1,633 (38%) females of Lu. spinicrassa dissected, 26 (1.6%) were infected with promastigotes, while no natural infection was found in 209 females of other species. The flagellates were identified as Leishmania braziliensis braziliensis using PCR with species specific primers derived from nuclear DNA and hybridization using species specific probe labelled with digoxigenin. This parasite had been previously isolated from patients with cutaneous leishmaniasis from the same area. These results show Lu. spinicrassa as a new proven vector of Leishmania braziliensis in the Andean region of Venezuela