Updates on Geographical Dispersion of Leishmania Parasites Causing Cutaneous Affections in Algeria

Abstract

Leishmaniases are neglected tropical diseases of public health concern in Algeria. To update the geographical distribution of Leishmania spp. causing cutaneous affection, we examined a set of Giemsa-stained smears prepared from skin lesions of the patients suspected to have cutaneous leishmaniasis (CL) in various geographical areas in Algeria. The identification of Leishmania parasites was performed using microscopy, conventional PCR, and PCR-RFLP (PCR-Restriction Fragment Length Polymorphism) targeting ITS1-rDNA. Among 32 smears provided from 27 suspected patients with cutaneous lesions, no trace of parasites was observed in the smear of three patients using microscopy and molecular approaches. Furthermore, four patients presented at least two lesions. PCR-RFLP confirmed the presence of Leishmania in 29 smears prepared from 24 patients. Two biopsies, negative after microscopic examination, were found positive by PCR. Of these 29 PCR positive smears (24 patients), 20 were identified using RFLP-PCR as L. major, two as L. tropica, and two as L. infantum. We found L. major infected patients from Ain skhouna, Biskra, El M'hir, Ghardaïa, M'Sila, and Saida, in agreement with previously reported cases. Furthermore, we highlighted for the first time, the identification of L. major in the patients from Bourkika, Bou Kremissa, Bou Saada Clef, Hajout, Maghnia, Médéa, Menaceur, Messad, Mostaghanem, Nador, Oran, and Sidi Okba. A phylogenetic reconstruction performed with sequences collected from the PCR products confirmed these identifications. Our data provide additional information on the geographical extension of CL caused by L. tropica and L. infantum in Algeria.

Keywords: Leishmania infantum; Leishmania major; Leishmania tropica; PCR–RFLP; cutaneous leishmaniasis.

Figure 1

Schematic representation of leishmaniasis endemic regions for L. major, L. tropica, and L. infantum, and the geographical origin of cutaneous samples processed in the present study (red points).

Figure 2

PCR–RFLP of cutaneous biopsies collected in Algeria. (A) Schematic representation of BsuR1 (HaeIII) cut sites in amplified fragments of ITS1-rDNA in Leishmania major, L. tropica, and L. infantum (CLC DNA Workbench 5.2 software); (B) Ethidium bromide-stained agarose gel of HaeIII digested PCR products of Leishmania species extracted from Giemsa stained smears. M: molecular marker (50 bp); Lanes 1–3: undigested reference strains of L. major, L. tropica, and L. infantum; Lanes 4–6: digested L. major, L. tropica, and L. infantum isolated from the patients; Lane 7: negative control. For L. tropica isolates, the 20 bp fragment could not be observed in an agarose gel electrophoresis. The 57 and 60 bp fragments could not be discriminated; only bands at 200 and 60 bp were indicative and distinguished after agarose gel electrophoresis.

Figure 3

Neighbor-joining (NJ) phylogenetic tree constructed based on ITS1-rDNA sequence of Leishmania samples analyzed in the present study (samples entitled AVC) and those collected in GenBank.