Development of GFP/RFP transfected <i>Leishmania</i> strains in sand flies.

<p>Infection rates and intensities of co-infections in three combinations of <i>L. donovani</i> strains in <i>L. longipalpis</i> (LO) and <i>P. perniciosus</i> (PE). A, LEM3804 plus LV9: in late stage infections (days 7 and 10) LEM3804 mostly over-grew LV9, in some females LV9 disappeared. B, Gebre-1 plus LV9: Gebre-1 grew better than LV9, in a high proportion of females LV9 was lost during late stage infections. C, Gebre-1 plus LEM4265: in late stage infections Gebre-1 mostly over-grew LEM4265, in a high proportion of females LEM4265 disappeared.</p

Numbers of single and co-infected sand flies examined by fluorescence microscopy.

<p>PBM, post bloodmeal;</p><p>Dis, number of dissected females;</p><p>Inf, number of females infected with at least one <i>Leishmania</i> strain;</p><p>B, number of females infected with both <i>Leishmania</i> strains;</p><p>H, number of females with hybrid promastigotes seen and photographed under fluorescence microscope;</p><p>-, not done.</p

Map of the spatial distribution of TcI isolates used in the study.

<p>Colours represent the different biomes where samples were obtained: Green: Amazon; Orange: Atlantic Forest; Red: Cerrado. Blue: Pantanal. Grey: Caatinga.</p

Unrooted neighbour-joining tree based on <i>D</i>_AS values between multilocus genotypes generated for 92 sylvatic Brazilian TcI strains.

<p><i>D</i>_AS values were calculated as the mean across 1000 random diploid re-samplings of the data set. Branch colors represent the biomes where samples were collected: Green, Amazon; orange, Atlantic Forest; red, Cerrado; Bblue, Pantanal; gray, Caatinga. Symbols correspond to host/vector species: Triangle, <i>Didelphis</i> spp.; square, <i>Chiroptera</i> spp.; circle, Triatomine; diamond, Primate; inverted triangle, rodent; pentagon, dog; hexagon, Raccoon.</p

Pairwise F_ST values for microsatellite data grouped according to the collection site.

<p>Pairwise F_ST values for microsatellite data grouped according to the collection site.</p

Population genetic parameters for <i>a priori</i> sylvatic populations of TcI in Brazil.

<p>Population genetic parameters for <i>a priori</i> sylvatic populations of TcI in Brazil.</p

Properties of six <i>T</i>. <i>cruzi</i> MLST loci.

<p>Properties of six <i>T</i>. <i>cruzi</i> MLST loci.</p

Maximum-Likelihood tree constructed from maxicircle sequences for 62 TcI Brazilian isolates.

<p>Colors represent the biomes where samples were obtained: Green, Amazon; orange, Atlantic Forest; red, Cerrado; blue, Pantanal; gray, reference sequences from Caatinga. Symbols correspond to strain host/vector: Triangle, <i>Didelphis</i> spp.; square, <i>Chiroptera</i> spp.; circle, triatomine; diamond, primate; inverted triangle, rodent.</p

Details of gene targets.

<p>*Gene fragments included in the 7 loci MLST scheme;</p>†<p>Gene fragments included in the reduced 4 loci MLST scheme;</p>a<p>Gene ID: GenBank access number for the complete gene in the CL-Brener strain;</p>b<p>Amplicon size refers to the sequence size of the gene fragment including the primers regions;</p>c<p>5′ starting position: indicates the position where the analyzed sequence starts, counting from the first base of the amplicon;</p>d<p>Fragment Length refers to the sequence length used for the analyses (the analyzed fragments do not include the primer regions).</p><p>Details of gene targets.</p

Neighbor Joining tree based on the concatenation of 7 selected MLST fragments: <i>Rb19</i>, <i>TcMPX</i>, <i>HMCOAR</i>, <i>RHO1</i>, <i>GPI</i>, <i>SODB</i> and <i>LAP</i>.

<p>Different DTUs are represented by vertical bars. Branch values represent bootstrap values (1000 replications). Heterozygous sites were considered as average states (see <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003117#s2" target="_blank">methods</a>). Scale bar at the bottom left represents uncorrected p-distances.</p