The natural infection of sandflies by Leishmania in wild-caught specimens was studied, using the polymerase chain reaction (PCR)-hybridization technique. The PCR was carried out using 2 oligonucleotides (primers 3J1 and 3J2) derived from a repetitive nuclear DNA sequence. The primers support the enzymatic amplification of a fragment of approximately 500 bp, present in the nuclear DNA of Leishmania braziliensis. The expected band was observed in 5 of 65 sandflies containing flagellates. After hybridization with a species-specific probe, we confirmed natural infection by L. braziliensis. The technique allowed the identification of Lutzomyia gomezi and Lu. panamensis as vectors of L. braziliensis in an endemic area of cutaneous leishmaniasis in Urama, Puerto Cabello district in Venezuela. As far as we are aware, this work constitutes the first report of natural infection of Lu. panamensis with L. braziliensis in the study area. We also demonstrate that PCR-hybridization is a suitable approach to establish the Leishmania-sandfly relationship and will be useful in epidemiological studies of leishmaniasis in endemic areas.

This report describes the isolation of a Leishmania chagasi strain from a bat (Carollia perspicillata), and its identification using biological methods and molecular characterization. The parasites were isolated in an artificial culture medium from a blood sample extracted from a bat heart. The isolate was then inoculated into the footpads of Balb/c mice, which subsequently developed a typical nodular leishmanial lesion; the parasites were confirmed as Leishmania by smear and histopathology. Molecular characterization of the parasites was performed by polymerase chain reaction with species-specific primers, kDNA restriction pattern following Hae III endonuclease digestion and dot blot hybridization using a kDNA probe. This report demonstrates that bats can be hosts for L. chagasi species and suggests the need for studies to determine whether they may be involved in foci of visceral leishmaniasis.

(courage, self-affirmation, autonomy) and negative (defeat, despair, and an inability to adapt).

This study reports cutaneous leishmaniasis (CL) caused by Leishmania chagasi/Le. infantum in Venezuela, with some atypical characteristics. Out of 63 cases of CL in the suburbs of Altagracia de Orituco, Guarico State, Venezuela, 30 presented clinical, parasitological, immunological and epidemiological features different from those of the classical CL known in the country. The initial lesion was small and nodular, which, if not treated, might progress to a superficial ulcer. No secondary infection was observed. The identification of the isolates was carried out by molecular techniques. Twelve species of phlebotomine sandflies were caught, the most abundant being Lutzomyia evansi and Lu. longipalpis s.l., known vectors of Le. chagasi/Le. infantum. The existence of Le. chagasi/Le. infantum and its vectors in an endemic area of CL has implications and we suggest that epidemiological studies should be carried out to obtain a clearer picture of the extent of this CL form in Venezuela.

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