Descriptive statistics revealing genetic characteristics and variation of the ten microsatellite loci detected in the population of 55 <i>L. major</i> strains isolated from Tunisian rodents and worldwide.

<p>N, number of genotypes; NA, number of allele per locus; Ho, observed heterozygosity; He, expected heterozygosity; Fis, inbreeding coefficient.</p><p>Descriptive statistics revealing genetic characteristics and variation of the ten microsatellite loci detected in the population of 55 <i>L. major</i> strains isolated from Tunisian rodents and worldwide.</p

Neighbour-joining tree inferred from the Dps distances calculated for 55 <i>L. major</i> strains isolated from different rodents (40 Tunisian and 15 from other geographic origins) according to the 10 microsatellites analyzed.

<p>Strains isolated among <i>P. obesus</i> (P), <i>M. shawi</i> (M), <i>Tatera sp.</i> (T) and <i>R. opimus</i> (R) were classified into 10 genotypes Lmj01, Lmj02, Lmj14, Lmj15, Lmj17, Lmj21, Lmj37, Lmj39, Lmj65 (as described in<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107043#pone.0107043-AlJawabreh1" target="_blank">[21]</a>) and RdTN from Africa (AF), Middle East (ME) and Central Asia (CA). RdTN indicates the genotype obtained from Tunisian reservoirs. Results are shown as radial tree where the percentages (under 80%) with which a branch is supported in 1000 bootstrap replications are indicated.</p

Theoretical sizes and repetition numbers among the ten-microsatellite loci in Tunisian studied isolates.

<p>*: New allele not previously described.</p><p>Theoretical sizes and repetition numbers among the ten-microsatellite loci in Tunisian studied isolates.</p

Geographical distribution of strains isolated from the Central Tunisia study area.

<p>Panel A shows the square delimited study area in the Governorate of Sidi Bouzid (in gray). Panel B represents the land satellite image of the study area showing the distribution of animal reservoir hosts from which the <i>L. major</i> strains were isolated. Isolates from <i>P. obesus</i> origin were noted P and colored in green, from <i>M. shawi</i> origin were noted Mer and colored in yellow, and from <i>M. nivalis</i> was noted M and colored in red. Spatial data related to the reservoir hosts of these strains were collected using the Global Positioning System (GPS). Satellite imagery: ArcGIS software.</p

Map of Tunisia and schematic illustration of gene flow between the study areas.

<p>(A1) Location of Tunisia, North Africa. (B1) Location of the Gafsa, Sidi Bouzid and Kairouan Governorates within Tunisia. (C1) Zoom of the area under study to show Chrarda and Mnara (Nasrallah delegation) (Mnara and Chrarda are shown in the map with red asterisks) in the Kairouan Governorate.</p

The unrooted neighbor-joining tree inferred from pair-wise Cavalli-Sforza and Edwards’ chord distances based on the 10 microsatellite data of 60 <i>Leishmania</i> major isolates (SBZ1991-1992 and SBZ2008-2012) shows that the historical isolates (SBZ1991-1992, n = 35; orange labels) and the recent isolates (SBZ2008-2012, n = 25; green labels) can be subdivided in five clusters.

<p>The unrooted neighbor-joining tree inferred from pair-wise Cavalli-Sforza and Edwards’ chord distances based on the 10 microsatellite data of 60 <i>Leishmania</i> major isolates (SBZ1991-1992 and SBZ2008-2012) shows that the historical isolates (SBZ1991-1992, n = 35; orange labels) and the recent isolates (SBZ2008-2012, n = 25; green labels) can be subdivided in five clusters.</p

Geographic sites, year of isolation and number of <i>L</i>. <i>major</i> isolates included in the study.

<p>Geographic sites, year of isolation and number of <i>L</i>. <i>major</i> isolates included in the study.</p

Estimated population structure of <i>L</i>. <i>major</i> in Tunisia as inferred by the STRUCTURE software on the basis of the data on 10 microsatellite markers obtained for 137 recent isolates from the Gafsa (GF2008-2012; n = 66), Kairouan (KR2008-2012; n = 46) and Sidi Bouzid (SBZ2008-2012; n = 25) governorates.

<p>(A) Plot of the mean posterior probability (LnP(D)) values per clusters (K), based on 10 replicates per K, generated by the STRUCTURE software, and (B) delta-K analysis of LnP(K). (C) STRUCTURE plots grouped by Q-matrix (estimated membership coefficient for each sample) showing the distribution of genetic variation (C.1) at K = 2 and (C.2) at K = 4. Each strain is represented by a vertical line, which is partitioned into the colored segments that represent the parasite estimated membership fractions in K. The same color indicates that the isolates belong to the same group. Different colors for the same isolate indicate the percentage of the genotype shared with each group. Gene flow between populations is indicated with arrows.</p

Genetic diversity indices, estimated from microsatellite data (10 loci), for the 172 <i>L</i>. <i>major</i> isolates analyzed in this study.

<p><i>L</i>. <i>major</i> isolates were included in the different subpopulations: historical (SBZ1991-1992) and recent populations (SBZ2008-2012) from the Sidi Bouzid governorate; recent populations from Gafsa (GF2008-2012), Kairouan (KR2008-2012) and Sidi Bouzid (SBZ2008-2012); the (KR2008-2012) population was further divided in samples from Mnara (MN2008-2012, historical epidemic focus) and from Chrarda (CH2008-2012, emerging epidemic focus).</p><p><i>A</i> = allelic richness per population based on the standardized minimal sample size; <i>H</i>s = gene diversity; <i>Ho</i> = observed heterozygosity; <i>He</i> = expected heterozygosity; <i>F</i>_IS = inbreeding coefficient.</p><p>Genetic diversity indices, estimated from microsatellite data (10 loci), for the 172 <i>L</i>. <i>major</i> isolates analyzed in this study.</p