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      Comparison of Leishmania killicki (syn. L. tropica) and Leishmania tropica Population Structure in Maghreb by Microsatellite Typing

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          Abstract

          Leishmania ( L.) killicki (syn. L. tropica), which causes cutaneous leishmaniasis in Maghreb, was recently described in this region and identified as a subpopulation of L. tropica. The present genetic analysis was conducted to explore the spatio-temporal distribution of L. killicki (syn. L. tropica) and its transmission dynamics. To better understand the evolution of this parasite, its population structure was then compared with that of L. tropica populations from Morocco. In total 198 samples including 85 L. killicki (syn. L. tropica) (from Tunisia, Algeria and Libya) and 113 L. tropica specimens (all from Morocco) were tested. Theses samples were composed of 168 Leishmania strains isolated from human skin lesions, 27 DNA samples from human skin lesion biopsies, two DNA samples from Ctenodactylus gundi bone marrow and one DNA sample from a Phlebotomus sergenti female. The sample was analyzed by using MultiLocus Enzyme Electrophoresis (MLEE) and MultiLocus Microsatellite Typing (MLMT) approaches. Analysis of the MLMT data support the hypothesis that L. killicki (syn. L. tropica) belongs to the L. tropica complex, despite its strong genetic differentiation, and that it emerged from this taxon by a founder effect. Moreover, it revealed a strong structuring in L. killicki (syn. L. tropica) between Tunisia and Algeria and within the different Tunisian regions, suggesting low dispersion of L. killicki (syn. L. tropica) in space and time. Comparison of the L. tropica (exclusively from Morocco) and L. killicki (syn. L. tropica) population structures revealed distinct genetic organizations, reflecting different epidemiological cycles.

          Author Summary

          Leishmania killicki (syn. L. tropica) was discovered in 1986. Few studies have been conducted on this parasite exclusively described in Maghreb. Consequently, many elements on its epidemiology, transmission, population structure and dynamics remain unknown.

          To better understand the evolution of this parasite, its population structure has been compared with that of L. tropica populations from Morocco using Multilocus Enzyme Electrophoresis (MLEE) and MultiLocus Microsatellite Typing (MLMT) typing. MLMT data support the hypothesis that L. killicki (syn. L. tropica) belongs to the L. tropica complex despite the strong genetic differentiation between them. Despite the probable recent divergence between L. killicki (syn. L. tropica) and L. tropica, they seem to evolve differently. Indeed, L. killicki (syn. L. tropica) appears slightly polymorphic and highly structured in space and time, while L. tropica was genetically heterogeneous, slightly structured geographically and temporally. The different population structures revealed distinct genetic organizations, reflecting different epidemiological cycles. Several parameters could explain these opposite epidemiological and genetic patterns such as ecosystems, vectors and reservoirs.

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          Most cited references20

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          Estimation of fixation indices and gene diversities.

          Considering the multinomial sampling of genotypes, unbiased estimators of various gene diversity measures in subdivided populations are presented. Using these quantities, formulae for estimating Wright's fixation indices (FIS, FIT, and FST) from a finite sample are developed.
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            Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification.

            The authors propose a new classification for the genus Leishmania Ross, 1903 based both on the use of intrinsic and extrinsic characters and on Linnean and Adansonian methods. The type of vertebrate host makes it possible to recognize the genus group: Leishmania designates Kinetoplastida parasites of mammals. Neighbouring forms which parasite reptiles are now grouped in the genus Sauroleishmania Ranque, 1973. Characteristics of the intravectorial cycle (supra- and peri-pyloric) are used to define the subgenus group (Leishmania, Viannia Lainson and Shaw, 1987). The classification uses biochemical, particularly enzymatic, characters. Elementary taxonomic units are made up of all the strains having the same isoenzyme profile, i.e. the zymodeme. The grouping of the zymodemes is usually performed through automatic techniques which lead to bush-like trees (dendrograms) showing either simple affinities between units (phenograms) or their phyletic relationships (cladograms). The branches recognized as being stable are individualized as "zymodeme complexes". They bear the name of either the previously defined species taxa or that of a specially created one. Two examples of taxonomic constructions, phenetic and cladistic, are presented. Finally, a general classification of the genus is proposed.
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              Microsatellite analysis reveals genetic structure of Leishmania tropica.

              The current rapid spread of leishmaniases caused by Leishmania tropica and the complexity of its clinical spectrum call for this parasite's epidemiological and evolutionary investigation. Evaluation of its population structure by isoenzyme electrophoresis and previous molecular biological analysis has proved difficult. In this study, we used 21 microsatellite loci to type 117 strains from different African and Asian locations. Eighty-one different genotypes were found. A genetic bottleneck supported by a gradient in the number of alleles and consistent with the geographical structure of the Middle East suggests an African origin of this species. A Bayesian approach identified 10 genetic clusters that correlated predominantly with geographical origin. The strains in the 'Asia' cluster form a very heterogeneous sub-population, with a varied but inter-related genotype that is geographically very widely dispersed and consistent with anthroponotic transmission of the parasite. The other nine clusters were more homogenous. The propagation of L. tropica appears to be predominantly clonal. In Africa and the Middle East, anthroponotic and zoonotic systems of distribution may contribute to the development of overlapping, genetically distinct populations of L. tropica.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS Negl Trop Dis
                PLoS Negl Trop Dis
                plos
                plosntds
                PLoS Neglected Tropical Diseases
                Public Library of Science (San Francisco, CA USA )
                1935-2727
                1935-2735
                8 December 2015
                December 2015
                : 9
                : 12
                : e0004204
                Affiliations
                [1 ]Laboratoire de Parasitologie-Mycologie Médicale et Moléculaire (code LR12ES08), Département de Biologie Clinique B, Faculté de Pharmacie, Université de Monastir, Tunisie
                [2 ]Centre National de Référence des Leishmanioses, Département de Parasitologie-Mycologie, CHRU de Montpellier, Université de Montpellier, Montpellier, France
                [3 ]UMR MIVEGEC (CNRS 5290-IRD 224-Université de Montpellier), Montpellier, France
                [4 ]College of Applied Medical Sciences, Clinical Laboratory Sciences Department, University of Hail, Hail, Saudi Arabia
                [5 ]Laboratoire d'éco-épidémiologie Parasitaire et Génétique des Populations. Institut Pasteur d'Algérie, Dély Ibrahim, Algeria
                US Food and Drug Administration, UNITED STATES
                Author notes

                The authors have declared that no competing interests exist.

                Conceived and designed the experiments: DC NH ZH JPD. Performed the experiments: DC LT PL HM. Analyzed the data: DC ALB FP. Contributed reagents/materials/analysis tools: ALB HB FP. Wrote the paper: DC ALB. Revised the paper: DC ALB NH JPD FP. Samples provider: ZH.

                Article
                PNTD-D-15-01097
                10.1371/journal.pntd.0004204
                4672892
                26645812
                52f77409-f3d5-4f56-9a27-2a7699408c8c
                © 2015 Chaara et al

                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 author and source are credited

                History
                : 23 June 2015
                : 9 October 2015
                Page count
                Figures: 3, Tables: 3, Pages: 13
                Funding
                This study was funded by the Institut Français de Coopération de Tunisie fellowship for PhD students. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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