Genetic diversity and structure in Leishmania infantum populations from southeastern Europe revealed by microsatellite analysis

Background: The dynamic re-emergence of visceral leishmaniasis (VL) in south Europe and the northward shift to
Leishmania-free European countries are well-documented. However, the epidemiology of VL due to Leishmania
infantum in southeastern (SE) Europe and the Balkans is inadequately examined. Herein, we aim to re-evaluate and
compare the population structure of L. infantum in SE and southwestern (SW) Europe.
Methods: Leishmania strains collected from humans and canines in Turkey, Cyprus, Bulgaria, Greece, Albania and
Croatia, were characterized by the K26-PCR assay and multilocus enzyme electrophoresis (MLEE). Genetic diversity
was assessed by multilocus microsatellite typing (MLMT) and MLM Types were analyzed by model- and
distance- based algorithms to infer the population structure of 128 L. infantum strains.
Results: L. infantum MON-1 was found predominant in SE Europe, whilst 16.8% of strains were MON-98. Distinct
genetic populations revealed clear differentiation between SE and SW European strains. Interestingly, Cypriot canine
isolates were genetically isolated and formed a monophyletic group, suggesting the constitution of a clonal MON-1
population circulating among dogs. In contrast, two highly heterogeneous populations enclosed all MON-1 and
MON-98 strains from the other SE European countries. Structure sub-clustering, phylogenetic and Splitstree analysis
also revealed two distinct Croatian subpopulations. A mosaic of evolutionary effects resulted in consecutive
sub-structuring, which indicated substantial differentiation and gene flow among strains of both zymodemes.
Conclusions: This is the first population genetic study of L. infantum in SE Europe and the Balkans. Our findings
demonstrate the differentiation between SE and SW European strains; revealing the partition of Croatian strains
between these populations and the genetic isolation of Cypriot strains. This mirrors the geographic position of
Croatia located in central Europe and the natural isolation of the island of Cyprus. We have analysed the largest
number of MON-98 strains so far. Our results indicate extensive gene flow, recombination and no differentiation
between MON-1 and MON-98 zymodemes. No correlation either to host specificity or place and year of strain
isolation was identified. Our findings may be associated with intensive host migration and common
eco-epidemiological characteristics in these countries and give valuable insight into the dynamics of VL.
* Correspondence:
1Laboratory of Molecular Parasitology, Hellenic Pasteur Institute, 127 Vasilissis
Sofias Avenus, 11521, Athens, Greece
Full list of author information is available at the end of the article
© 2013 Gouzelou et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative
Commons Attribution License (, which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
Gouzelou et al. Parasites & Vectors 2013, 6:342


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