Monthly human biting rates of and M and S molecular forms, from July to December 2004, in Dielmo, Senegal

<p><b>Copyright information:</b></p><p>Taken from "Dynamics of transmission of by and the molecular forms M and S of in Dielmo, Senegal"</p><p>http://www.malariajournal.com/content/7/1/136</p><p>Malaria Journal 2008;7():136-136.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2515330.</p><p></p

The nocturnal biting cycle of and M and S molecular forms, from July to December 2004, in Dielmo, Senegal

<p><b>Copyright information:</b></p><p>Taken from "Dynamics of transmission of by and the molecular forms M and S of in Dielmo, Senegal"</p><p>http://www.malariajournal.com/content/7/1/136</p><p>Malaria Journal 2008;7():136-136.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2515330.</p><p></p

Frequency distribution of <i>Pfcrt</i> intron 4 microsatellite types by codons 72–76 and 220 haplotype.

<p>247 isolates were typed (19, 23, 22, 33, 28, 22, 29, 17, 39 and 15 isolates in 1990, 1991, 1992, 1993, 1994, 1995, 1996, 1997, 1998 and 1999, respectively) for the intron 4 microsatellite by gene sequencing (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#s2" target="_blank">Materials and Methods</a>). There were 31 CVIETS haplotypes and 216 wild type haplotypes. The haplotype codes are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#pone.0000139.s002" target="_blank">Table S2</a>.</p

Temporal distribution of CQ and SP drug pressure and drug resistance in Dielmo in 1990–9.

<div><p>The drug pressure is expressed as No of treatments/person/year (first graph) and as overall No of treatment courses administered per year (second graph). Panels A and B refer to CQ and SP, respectively. The prevalence of the <i>Pfcrt</i> mutant alleles was calculated from molecular beacon studies (N = 324) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#pone-0000139-g002" target="_blank">Figure 2</a>), while the prevalence of the <i>Pfdhfr-ts</i> triple mutant was calculated from the full gene sequences available (N = 202) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#pone-0000139-g003" target="_blank">Figure 3</a>). <i>In vitro</i> susceptibility assays were carried out in 1990–4 during the rainy season (N = 26) and from 1995 onwards for the last 2–3 months of the year, namely from 7/11/1995–26/12/1995 (N = 46) ; 6/01/1996–3/12/1996 (N = 59); 27/10/97–15/121997 (N = 26) ; 10/01/1998–15/11/1998 (N = 54) and 29/09/1999–08/11/1999 (N = 25). The proportion of interpretable CQ and pyrimethamine susceptibility tests was 68–81% and 72–81%, respectively, depending on the year. The prevalence of resistance is expressed as the percent of interpretable assays presenting a CI₅₀ for CQ >100 nM or a CI₅₀ for pyrimethamine>2000 nM. The occurrence of a second clinical malaria episode within 7, 14, 21 and 28 days of treatment was calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#s2" target="_blank">Materials and Methods</a>. The bars correspond to the 95% confidence interval. The years before implementation of CQ and SP (1990–4) are grouped together.</p> <p> A. CQ pressure, <i>Pfcrt</i> 76T resistance mutation, CQ <i>in vitro</i> resistance and prevalence of clinical attacks following a CQ treatment</p> <p> B. SP pressure, <i>Pfdhfr-ts</i> triple mutant, pyrimethamine <i>in vitro</i> resistance and prevalence of clinical attacks following a SP treatment</p> <p>Colour codes: 1990–4: grey; 1995: purple; 1996: yellow; 1997: light green; 1998: light blue; 1999: orange.</p></div

Frequency distribution of <i>Pfdhfr-ts</i> −4.4 kb/−01 kb+0.5 kbmicrosatellite haplotypes by coding the sequence mutant type.

<p>The microsatellites were determined as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#s2" target="_blank">Materials and Methods</a> for 81 isolates. The haplotype codes are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#pone.0000139.s003" target="_blank">Table S3</a>.</p

Temporal variation of the relative <i>Pfdhfr-ts</i> gene polymorphism in Dielmo during 1990–99.

<p>The 1.8 kb PCR fragment corresponding to the full length <i>Pfdhfr-ts</i> coding sequence was sequenced on both strands for a total of 204 isolates. The yearly distribution of the various genotypes is shown, using the colour code shown to the right of the figure. The alleles presenting synonymous mutations were omitted from the colour coding. The C59Y non-synonymous substitution was a TGT to TAT mutation. The synonymous mutations (CTA to TTA or CTC for codon 40, GGA to GGC for codon 241) are not depicted. No mutation was detected at codon 16 and 164, and no <i>bolivia</i> repeat type was observed. Overall there were 155 isolates with wild type coding sequence and 49 isolates with non synonymous mutations (76% and 24%, respectively). There were 15 (7.3%) single mutant<i>s</i> [51I (0.5%), 59Y (1%), 108N (5.8%)], one (0.5%) 51I 108N double mutant and 33 (16.2%) 51I 59R 108N triple mutants.</p

Temporal variation of the multiplicity of infection in Dielmo (A), frequency of Pfcrt codon 76 and <i>Pfdhfr-ts</i> codon 108 genotypes (B) and frequency of infections with only mutant type detected (C).

<div><p>The number of isolates typed at each locus is indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000139#pone-0000139-t001" target="_blank">Table 1</a>. (A). Multiplicity of infection is depicted separately for each locus. For <i>Pfmsp1</i> block2, the figures derive from nested PCR analysis using family-specific primers and allele identification based on allelic family assignment and size polymorphism. For <i>Pfcrt</i> and <i>Pfdfhr-ts</i>, it is based on K76T and S108N genotypes determined by molecular beacons, respectively. Symbols used: (Red triangles): <i>Pfcrt</i> codon 76 genotype; (green squares): <i>Pfdhr-ts</i> codon 108 genotype, (blue open circles) <i>Pfmsp1</i> block2.</p> <p>B) Allelic frequency of resistance genotypes, calculated as percentage of mutant genotype within the total number of alleles detected for each locus. Symbols used as in A.</p> <p>C) Percentage of isolates containing only the mutant type. Symbols used as in A.</p></div