Abstract
Leishmania infantum causes Visceral and cutaneous leishmaniasis in northern Morocco. It predominantly affects children under 5 years with incidence of 150 cases/year. Genetic variability and population structure have been investigated for 33 strains isolated from infected dogs and humans in Morocco. A multilocus microsatellite typing (MLMT) approach was used in which a MLMtype based on size variation in 14 independent microsatellite markers was compiled for each strain. MLMT profiles of 10 Tunisian, 10 Algerian and 21 European strains which belonged to zymodeme MON-1 and non-MON-1 according to multilocus enzyme electrophoresis (MLEE) were included for comparison. A Bayesian model-based approach and phylogenetic analysis inferred two L.infantum sub-populations; Sub-population A consists of 13 Moroccan strains grouped with all European strains of MON-1 type; and sub-population B consists of 15 Moroccan strains grouped with the Tunisian and Algerian MON-1 strains. Theses sub-populations were significantly different from each other and from the Tunisian, Algerian and European non MON-1 strains which constructed one separate population. The presence of these two sub-populations co-existing in Moroccan endemics suggests multiple introduction of L. infantum from/to Morocco; (1) Introduction from/to the neighboring North African countries, (2) Introduction from/to the Europe. These scenarios are supported by the presence of sub-population B and sub-population A respectively. Gene flow was noticed between sub-populations A and B. Five strains showed mixed A/B genotypes indicating possible recombination between the two populations. MLMT has proven to be a powerful tool for eco-epidemiological and population genetic investigations of Leishmania.
Highlights
Leishmaniasis constitutes a group of diseases caused by obligatory, intracellular, protozoan parasites of the genus Leishmania, that cause a spectrum of diseases, ranging from self-limiting, self-curing cutaneous leishmaniasis (CL) to visceral leishmaniasis (VL) with fatal spontaneous evolution [1]
A panel of 14 microsatellite markers was applied to investigate the genetic polymorphism and population structure of Moroccan L. infantum strains and to compare them with strains from North African and European countries. Both phylogenetic analysis based on genetic distances as well as Structure analysis using a Bayesian clustering approach grouped the Moroccan strains together with MON-1 strains or strains closely related to MON-1 from Tunisia, Algeria and Europe in Population 1
Isoenzyme analysis had been performed for only two of the Moroccan strains studied which were typed as MON-1
Summary
Leishmaniasis constitutes a group of diseases caused by obligatory, intracellular, protozoan parasites of the genus Leishmania, that cause a spectrum of diseases, ranging from self-limiting, self-curing cutaneous leishmaniasis (CL) to visceral leishmaniasis (VL) with fatal spontaneous evolution [1]. More than 90% of global VL cases occur in just six countries: India, Bangladesh, Sudan, South Sudan, Brazil and Ethiopia [2].Most strains of L. infantum isolated from Mediterranean foci belong to the predominant zymodeme MON-1 [3], despite their very wide geographical distribution. Different methods have been used for the identification and classification of Leishmania parasites as reviewed [5] These methods, including multilocus enzyme electrophoresis (MLEE) which is still considered as the gold standard for species and strain typing of Leishmania, are limited in the intrinsic level of polymorphism they can detect and are, only in exceptional cases, able to differentiate strains in the zymodeme MON-1 [5]. We used the above mentioned 14 microsatellites markers to investigate the genetic polymorphism and population structure of Moroccan L. infantum strains isolated from human and canine cases (CanL). This comparison was done to understand VL transmissions scenarios in North Africa and South Europe
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