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APPLICATION OF MICROSATELLITE GENOTYPING TO THE STUDY OF A RESTRICTED LEISHMANIA INFANTUM FOCUS: DIFFERENT GENOTYPE COMPOSITIONS IN ISOLATES FROM DOGS AND SAND FLIES

Leishmania infantum polymorphism was studied by DNA microsatellite analysis of 110 L. infantum stocks (94 from dogs, 15 from sand flies, and 1 from a human visceral case) from a rural leishmaniasis-endemic area (Priorat) in northeastern Spain. Three microsatellites of the eight present in three fragments (internal transcribed spacer, Lm4, and Lm2) of L. infantum nuclear DNA are polymorphic inside the focus, resulting in 17 genotypes. Isolates from dogs and sand flies had different allelic compositions and shared only four genotypes. Microsatellite analysis is useful for L. infantum genotyping and epidemiologic tracking. Its application with strains from dogs and vectors in an area endemic for leishmaniasis shows the heterogeneous distribution of L. infantum in hosts living in sympatric conditions.

Received June 21, 2006. Accepted for publication December 14, 2006.

Acknowledgments: Many of the Leishmania stocks studied were isolated during summer campaigns in collaboration with researchers from the Laboratoire d’Écologie Médicale (Montpellier, France). We acknowledge their help, particularly the assistance of Drs. E. Guilvard, G. Moreno and F. Pratlong in the isolation of stocks from sand flies and isoenzyme identification. We thank Drs. J. G. Lorén, M. C. Fusté, and M. Farfán (Laboratory of Microbiology, Facultat de Farmàcia, Universitat de Barcelona, Barcelona) for genetic analysis; and Dr. G. Cuny (Laboratoire de Recherches et de Coordination sur les Trypanosomes, Montpellier, France) and Dr. P. Darlu (Laboratoire de Génétique Épidémiologique et Structure des Populations Humaines, Villejuif, France) for valuable comments and suggestions. We also thank the inhabitants of the Priorat region for their collaboration in fieldwork, S. Tebar for excellent technical assistance, R. Rycroft for correcting the English version of the manuscript, and the personnel at the Serveis Cientificotècnics, Universitat de Barcelona for DNA sequencing, assistance, and technical advice.

Financial support: This study was supported by the Spanish Government (PB86-0546, SAL-90-0960-CO2, PB94-0865, FIS 99/0036-01, FIS 01/0831, and SAF02-10172) and the European Union (TS2M-0058-F and FAIR 6-CT98-4104).

^* Address correspondence to Roser Fisa, Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avenida Joan XXIII s/n, 08028 Barcelona, Spain. E-mail: rfisa{at}ub.edu

Authors’ addresses: Liliana Montoya, Montserrat Gállego, Montserrat Portús, and Roser Fisa, Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avenida Joan XXIII s/n, 08028 Barcelona, Spain, Telephone: 34-93-4024502, Fax: 34-93-4024504. Béatrice Gavignet and Renaud Piarroux, Santé Environnement Rural Franche-Comté, Université de Franche-Comté, Besançon, France, Telephone: 33-38-166-8068. Jean-Antoine Rioux, Institut de Botanique, Université Montpellier I, Montpellier, France.