The Plurinational State of Bolivia is one of the Latin American countries with the highest prevalence of leishmaniasis, highlighting the lowlands of the Department of La Paz where about 50% of the total cases were reported. The control of the disease can be seriously compromised by the intrinsic variability of the circulating species that may limit the efficacy of treatment while favoring the emergence of resistance. Fifty-five isolates of Leishmania from cutaneous and mucocutaneous lesions from patients living in different provinces of the Department of La Paz were tested. Molecular characterization of isolates was carried out by 3 classical markers: the rRNA internal transcribed spacer 1 (ITS-1), the heat shock protein 70 (HSP70) and the mitochondrial cytochrome b (Cyt-b). These markers were amplified by PCR and their products digested by the restriction endonuclease enzymes AseI and HaeIII followed by subsequent sequencing of Cyt-b gene and ITS-1 region for subsequent phylogenetic analysis. The combined use of these 3 markers allowed us to assign 36 isolates (65.5%) to the complex Leishmania (Viannia) braziliensis, 4 isolates (7, 27%) to L. (Viannia) lainsoni. and the remaining 15 isolates (23.7%) to a local variant of L. (Leishmania) mexicana. Concerning in vitro drug susceptibility the amastigotes from all isolates where highly sensitive to Fungizone® (mean IC50 between 0.23 and 0.5 μg/mL) whereas against Glucantime® the sensitivity was moderate (mean IC50 ranging from 50.84 μg/mL for L. (V.) braziliensis to 18.23 μg/mL for L. (L.) mexicana. L. (V.) lainsoni was not sensitive to Glucantime®. The susceptibility to miltefosine was highly variable among species isolates, being L. (L.) mexicana the most sensitive, followed by L. (V.) braziliensis and L. (V.) lainsoni (mean IC50 of 8.24 μg/mL, 17.85 μg/mL and 23.28 μg/mL, respectively).
Bibliographical noteFunding Information:
We are grateful to the National Institute Laboratory of Health (INLASA) of La Paz (Bolivia) for the donation of 55 Leishmania Bolivian isolates and to Drs. M. Portús Vinyeta (Universidad de Barcelona), Dr. A. Torraño and Dr. M. Domínguez (Instituto Carlos III, Madrid), Dr. Cesar Ramírez (Universidad Javeriana, Colombia) and Dr. Lilian Yépez-Mulia (Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Ciudad de México, Mexico) for providing reference strains. This study was supported by the Spanish Agency of International Cooperation for Development (AECID) through projects No.s A/024457/09, A/030160/10 and AP/0397670/11. P.E.B.R. was a recipient of a Pre-Doctoral fellowship from AECID.
- Bolivian isolates
- Cytochrome b
- Drug susceptibility
- Molecular characterization