KDNA signatures of the 447-bp minicircle fragment of <i>L. infantum</i> (<i> = L. chagasi</i>) strains isolated from human patients.

<p>Five µ l of the LSSP-PCR reaction products, were loaded in each lane of a 6% polyacrylamide gel and silver stained. Lanes 2 to 17: genotype I (lanes 2–4: HP7, HB5 and HB9); genotype II (lanes 5–6: HP10 and HP1); genotype III (lanes 7–8: HP6 and HB6); genotype VI (lane 9: HP5); genotype VII (lane 10: HB8); genotype VIII (lane 11: HP3); genotype IX (lane 12: HP8); genotype IV (lanes 13–14: HB4 and HB9); genotype V (lanes 15–16: HP4 and PP75); lane 17 the negative control of the LSSP-PCR reaction. Migration of the markers of the 1 Kb DNA ladder (Life Technologies, Inc., Gaithersburg, MD) is shown in lane 1, with the following molecular sizes (from the bottom up): 75, 134, 154, 201, 220, 298, 344, 396, 506, 517, 1018 and 1636 bp. Genotypes are indicated by roman numbers.</p

<i>Leishmania infantum</i> strains used in this study.

#<p>Parasite strains isolated from Belo Horizonte, Minas Gerais State, Brazil;</p>*<p>Strains isolated from the Lisbon Metropolitan Region (LMR), Portugal;</p>§<p>KDNA genotypes were determined by LSSP – PCR technique by using the MC1 primer as the driver.</p

KDNA signatures of the 447-bp minicircle fragment of <i>L. infantum</i> (<i> = L. chagasi</i>) strains isolated from the canine reservoir.

<p>Five µ l of the LSSP-PCR reaction products, were loaded in each lane of a 6% polyacrylamide gel and silver stained. Lanes 2 to 11: genotype I (lanes 2–3: CB2 and CP3); genotype II (lane 4: CP5); genotype III (lanes 5–6: CB7 and CP2); genotype IV (lane 7: CP6); genotype V (lane 8: CB3); genotype VI (lane 9: IPT1); genotype VII (lane 10: CB10); genotype VIII (lane 11: CB4). Migration of the markers of the 1 Kb DNA ladder (Life Technologies, Inc., Gaithersburg, MD) is shown in lane 1, with the following molecular sizes (from the bottom up): 75, 134, 154, 201, 220, 298, 344, 396, 506, 517, 1018 and 1636 bp. Genotypes are indicated by roman numbers.</p

Dendogram obtained by analysis of LSSP-PCR profiles of 40 <i>Leishmania infantum</i> (<i> = L. chagasi</i>) strains isolated from VL human cases or canine reservoirs.

<p>Genetic distances obtained by LSSP-PCR analysis of the 447 Pbp kDNA minicircle. The main constitutive bands of the LSSP-PCR profiles were used to build the phenetic tree by using the UPGMA method. Strains belonged to the same kDNA genotype are indicated on the right.</p

Additional file 1: Table S1. of Studies in the mouse model identify strain variability as a major determinant of disease outcome in Leishmania infantum infection

Primers used to amplify mouse genes. (PDF 8 kb

Characterization of the 4 populations found by STRUCTURE (<i>K</i> = 4) analysis of 139 strains of Mediterranean <i>L. infantum</i>

a<p>The two strains with mixed MON-1/non-MON-1 alleles (INF-32, PT2(I)) have been excluded. INF-35 (MON-108) is part of the MON-1 populations. In case of ES9(III) and ES10(III) only the non-MON-1 genotype has been considered, as these strains have been identified as MON-24 by MLEE.</p>b<p>3 strains from Madrid (ES1(I), ES3(I), ES6(I)) originally identified as MON-1 clearly group with the non-MON-1 population and have been considered as part of the non-MON-1 population 4. The first number in the bracket indicates the number of strains from a given focus belonging to the respective population, the second one the overall number of strains from a given focus. N, number of strains; P, proportion of polymorphic loci; MNA, mean number of alleles; <i>H</i>_o, observed heterozygosity; <i>H</i>_e, expected heterozygosity; <i>F</i>_IS, inbreeding coefficient; NA_u, number of unique alleles; I_A-Index of association.</p

Allele-frequencies of each of the 14 microsatellites studied for 113 strains of <i>L. infantum</i> MON-1 and 26 strains of non-MON-1

a<p>Prominent alleles (>10%) are marked as bold numbers.</p>b<p>Two strains of mixed MON-1/non-MON-1 alleles have been excluded (INF-32 and PT2(I)). For ES9(III) and ES10(III) only the non-MON-1 genotype has been considered, as these strains have been identified as MON-24 by MLEE.</p

MLST-profiles of strains of mixed ancestry (heterozygous and mosaic genotypes) and alleles unique to Alto Douro

a<p><i>Phlebotomus</i> isolates;</p>b<p>Alleles unique for strains from Alto Douro; bold: alleles found exclusively in non-MON-1 strains, italics: alleles found exclusively in MON-1 strains Several alleles are found in both groups-MON-1 and non-MON-1, sometimes with similar frequencies, in other cases the frequency is different in MON-1 and non-MON-1, up to the case that some alleles are dominating in one of those groups-see <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000261#pntd-0000261-t005" target="_blank">Table 5</a>. nd-not defined.</p

Designation and characteristics of <i>Leishmania infantum</i> strains used in this study

a<p>VL-Visceral leishmaniasis, CL-Cutaneous leishmaniasis, VCL-Viscero-cutaneous leishmaniasis, CanL-Canine leishmaniasis;</p>b<p>Metropolitan Region of Lisbon;</p>R<p>WHO-reference strain; n.d. not defined.</p

Factorial correspondence analysis (FCA) of <i>L. infantum</i> strains from the Mediterranean region.

<p>(A) All MON-1 and non-MON-1 strains included (141 strains). (B) Analysis of the three MON-1 populations (113 strains). TR–Turkey, TN–Tunisia, IL–Israel; Population 1 (Greece)–yellow squares, population 2 (Majorca+Ibiza)–blue squares, population 3 (mainland Spain+Portugal)–white squares, population 4 (non-MON-1)–grey squares.</p