Distance in km between the five study sites.

<p>Distance in km between the five study sites.</p

Mutations and corresponding amino acid substitutions in <i>pvceltos</i> gene.

<p>Mutations and corresponding amino acid substitutions in <i>pvceltos</i> gene.</p

Genetic differentiation between samples from Brazil, measured by pairwise <i>F</i>_ST values.

<p>Genetic differentiation between samples from Brazil, measured by pairwise <i>F</i>_ST values.</p

Modeling structure and <i>In silico</i> analysis of PvCelTOS.

<p>(A) Synonymous and non-synonymous mutations were illustrated by blue title and red title, respectively, on 3D structure of PvCelTOS. The red and blue clouds represent the negative and positive surface, respectively. (B) Synonymous and non-synonymous mutations found in our population and other described mutations are illustrated by blue bars and red bars, respectively, on PvCelTOS structure. The blue lines represent predicted linear B-cell epitopes and the red lines represent predicted T_CD8+ epitopes. On both (B and T predicted epitopes) the letter and number of each epitope indicate the C-terminal and N-terminal amino acid. The BepiPred values represent the predicted score of linear B-cell epitope in wild type haplotype (H1) and mutate strain (red number). The IEDB MHC-I indicates the mean binding prediction score of T_CD8+ epitopes and respective HLA binding frequency among 27 evaluated HLA. No differences of prediction T-cell epitopes are observed between wild types or mutate PvCelTOS.</p

Geographical map showing the five study sites and the respective Annual Parasitic Incidence (API) (SIVEP-Malaria).

<p>Geographical map showing the five study sites and the respective Annual Parasitic Incidence (API) (SIVEP-Malaria).</p

Alignment of protein sequences and frequency of mutations on field isolates.

<p>(A) CelTOS protein derivatives from <i>P</i>. <i>vivax</i> genome. The yellow mers represent the reference strain and the red mers the described mutate amino acid. (B) Frequencies of mutations in isolates from different Brazilian Amazon regions, where the colors blue, red, yellow and green represent genome identical to strains Sal-1; Brazil I and mutant isolates Ser10-Leu118 and Ser10-Val118, respectively.</p

PCR amplification of the <i>pvceltos</i> gene.

<p>Fig 2 shows in <b>Lane 1</b>: 100 bp Molecular Marker; <b>Lane 2</b>: Negative control (water); <b>Lane 3</b>: <i>In vitro</i> culture of <i>P</i>. <i>falciparum</i> (amplification with PvCelTOS primers); <b>Lane 4</b>: <i>In vitro</i> culture of <i>P</i>. <i>falciparum</i> (amplification with p126 primers); <b>Lane 5</b>: PCR positive control (<i>P</i>. <i>vivax</i>-infected sample); <b>Lanes 6, 7 and 8</b>: samples; <b>Lane 9</b>: 100 bp Molecular Marker.</p

Distribution of PvCelTOS haplotypes among five studied localities of Brazilian Amazon.

<p>Distribution of PvCelTOS haplotypes among five studied localities of Brazilian Amazon.</p

Comparison of genetic diversity among isolates from Brazil.

<p>Comparison of genetic diversity among isolates from Brazil.</p

Analysis of genetic diversity of PvCelTOS in <i>Plasmodium vivax</i> isolates.

<p>(A) Fig A represents four mutations in specific bp positions (24, 28, 109 and 352). (B) The graphic represents the frequency of mutations in isolates from each studied locality. The black bar indicates the synonymous mutation C24A; the gray bar, the non-synonymous mutations G28A; the striped bar, the synonymous mutation C109A and the white dotted bar represents the non-synonymous G352C. (*) Indicates that the differences between the frequency of SNP C109A was higher than that of other mutations by exact test and (+) indicates that the frequency of SNP G352C was higher than the frequency of SNPs C24A and G28A by Fisher’s. (*): p<0.05; (**): p<0.01; (***): p<0.0001.</p