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Table S2. Meteorological information for the region of Segou, 2004 to 2017. (DOCX 19 kb)

Table S1. Number of collected sand flies per household per month before and after IRS in the study area. Before IRS/LLINs, the number of collected flies per household ranged from 205 to 649 with a median of 293.00 and variance of 19778.94. After IRS/LLINs, the number of collected flies per household ranged from 25 to 159 with a median of 59.00 and variance of 3123.29. (DOCX 16 kb)

In this study, a sensitive and simple flow-injection chemiluminescence (CL) method was developed for the quantitative analysis of haemoglobin. The method is based on the ability of haemoglobin to enhance the CL signal generated by a H(2)O(2) -K(4) Fe(CN)(6) -fluorescein alkaline system enhanced by CdTe quantum dots. Under the optimized conditions, haemoglobin can be detected in concentration range 7.35 × 10(-9) -2.5 × 10(-6) mol/L, with a detection limit (3σ) of 1.8 × 10(-9) mol/L and a relative standard deviation (RSD; for 5 × 10(-7) mol/L haemoglobin) of 2.06% (n = 11). The present CL method was successfully applied for the determination of haemoglobin in three kinds of blood samples taken from an infant, an adult man, an adult woman and two reference samples. Compared with previous reports, the CL method described in this work is simple and rapid, with high sensitivity.

Additional file 1: Figure S1. Estimated gametocyte nucleic acid quantities in each reaction step based on highest dilution of gametocyte standard curve (103 gametocytes/μL). For dilution series used to assign gametocyte levels to field samples, the RNA stabilising buffer was L6 Buffer (450 μL) for the Mali study and RNAprotect (250 μL) for the Burkina Faso study.

Genome wide total heterozygosity and relative heterozygosity for founder colony Fd09. Plots depict heterozygosity measures for colony Fd09 across all chromosomes. Total pooled heterozygosity (Hp) was calculated in a sliding 10 kb window along the chromosome within the Fd09 colony. Dots indicate minimum and maximum values for a 1 Mb window, the black line indicates the average heterozygosity and the gray shading represents the standard deviation of total pooled heterozygosity across a 1 Mb window. Relative diversity (HpR) per 1 Mb window, calculated as the proportion of heterozygosity in a given pool relative to total heterozygosity within the source Fd09 colony. Color of point indicates per window elevated heterozygosity (blue), or reduced heterozygosity (red), plotted as log base 10 of the relative diversity. Phenotype pool identity is indicated in the lower left of each panel (high, low, zero oocysts). A relative heterozygosity value of 1 indicates the same heterozygosity levels in tested pool as compared to all other pools, values greater than 1 indicate greater heterozygosity in the tested pool and values less than 1 indicate lower heterozygosity in the tested pool. Given the log scale values of 0.5 and 2.0 are equidistant from 1. Candidate loci 9.1 and 9.2 are indicated by dark blue and light blue shading, respectively, on Chromosome 2. At locus 9.1, relative heterozygosity is decreased in the high pool and simultaneously reduced in the low and zero pools, while at locus 9.2, relative heterozygosity in increased in the high pool and decreased in the other pools. Each pool was comprised of DNA from 20 individual mosquitoes. (PDF 4973 kb)

In this study, a sensitive and simple flow-injection chemiluminescence (CL) method was developed for the quantitative analysis of haemoglobin. The method is based on the ability of haemoglobin to enhance the CL signal generated by a H(2)O(2) -K(4) Fe(CN)(6) -fluorescein alkaline system enhanced by CdTe quantum dots. Under the optimized conditions, haemoglobin can be detected in concentration range 7.35 × 10(-9) -2.5 × 10(-6) mol/L, with a detection limit (3σ) of 1.8 × 10(-9) mol/L and a relative standard deviation (RSD; for 5 × 10(-7) mol/L haemoglobin) of 2.06% (n = 11). The present CL method was successfully applied for the determination of haemoglobin in three kinds of blood samples taken from an infant, an adult man, an adult woman and two reference samples. Compared with previous reports, the CL method described in this work is simple and rapid, with high sensitivity.

Genome wide total heterozygosity and relative heterozygosity for founder colony Fd03. Plots depict heterozygosity measures for colony Fd03 across all chromosomes. Total pooled heterozygosity (Hp) was calculated in a sliding 10 kb window along the chromosome within the Fd03 colony. Dots indicate minimum and maximum values for a 1 Mb window, the black line indicates the average heterozygosity and the gray shading represents the standard deviation of total pooled heterozygosity across a 1 Mb window. Relative diversity (HpR) per 1 Mb window, calculated as the proportion of heterozygosity in a given pool relative to total heterozygosity within the source Fd03 colony. Color of point indicates per window elevated heterozygosity (blue), or reduced heterozygosity (red), plotted as log base 10 of the relative diversity. Phenotype pool identity is indicated in the lower left of each panel (high, low, zero oocysts). A relative heterozygosity value of 1 indicates the same heterozygosity levels in tested pool as compared to all other pools, values greater than 1 indicate greater heterozygosity in the tested pool and values less than 1 indicate lower heterozygosity in the tested pool. Given the log scale values of 0.5 and 2.0 are equidistant from 1. Candidate locus 3.1 is indicated by the red vertical shaded bar at coordinates 17.4-19.1 Mb. In this interval, relative heterozygosity is increased in the high pool and simultaneously reduced in the low and zero pools. Pools were comprised of DNA from 20 (zero oocyst pool), 17 (low oocyst pool), or 14 (high oocyst pool) individuals. (PDF 4909 kb)

Genome wide total heterozygosity and relative heterozygosity for founder colony Fd03. Plots depict heterozygosity measures for colony Fd03 across all chromosomes. Total pooled heterozygosity (Hp) was calculated in a sliding 10 kb window along the chromosome within the Fd03 colony. Dots indicate minimum and maximum values for a 1 Mb window, the black line indicates the average heterozygosity and the gray shading represents the standard deviation of total pooled heterozygosity across a 1 Mb window. Relative diversity (HpR) per 1 Mb window, calculated as the proportion of heterozygosity in a given pool relative to total heterozygosity within the source Fd03 colony. Color of point indicates per window elevated heterozygosity (blue), or reduced heterozygosity (red), plotted as log base 10 of the relative diversity. Phenotype pool identity is indicated in the lower left of each panel (high, low, zero oocysts). A relative heterozygosity value of 1 indicates the same heterozygosity levels in tested pool as compared to all other pools, values greater than 1 indicate greater heterozygosity in the tested pool and values less than 1 indicate lower heterozygosity in the tested pool. Given the log scale values of 0.5 and 2.0 are equidistant from 1. Candidate locus 3.1 is indicated by the red vertical shaded bar at coordinates 17.4-19.1 Mb. In this interval, relative heterozygosity is increased in the high pool and simultaneously reduced in the low and zero pools. Pools were comprised of DNA from 20 (zero oocyst pool), 17 (low oocyst pool), or 14 (high oocyst pool) individuals. (PDF 4909 kb)

Manhattan plot for colony Fd03. Results are given for the two replicates individually and the values combined by Fisher’s method. All genotyped loci were tested for association with two phenotypes by logistic regression using PLINK. Infection prevalence (blue) was defined as having more than one oocyst in the dissected midgut, infection intensity (red, measured only for mosquitoes carrying ≥1 oocyst) as having >5 oocysts. Dashed line represents a 0.01 p-value (i.e., 1/p = 100). Genes in the regions are shown below; Toll11 and Toll10 are the two rightmost genes on the positive strand. The combined plot represents values from reps 1 and 2 combined by the method of Fisher [47]. (PDF 127 kb)

Manhattan plot for colony Fd03. Results are given for the two replicates individually and the values combined by Fisher’s method. All genotyped loci were tested for association with two phenotypes by logistic regression using PLINK. Infection prevalence (blue) was defined as having more than one oocyst in the dissected midgut, infection intensity (red, measured only for mosquitoes carrying ≥1 oocyst) as having >5 oocysts. Dashed line represents a 0.01 p-value (i.e., 1/p = 100). Genes in the regions are shown below; Toll11 and Toll10 are the two rightmost genes on the positive strand. The combined plot represents values from reps 1 and 2 combined by the method of Fisher [47]. (PDF 127 kb)

Concordance of allele frequencies as called from pooled sequence and Sequenom individual genotyping. Concordance was assessed with data from three Sequenom plexes spread across all 3 candidate loci, 3.1, 9.1, and 9.2. SNPs were chosen independently of the phenotype group; that is, they were chosen on the basis of differing allele frequencies in either of the founder pools, and not for high allele differences between phenotype pools. Allele frequencies for Sequenom data were calculated from genotype calls and allele frequencies from pool sequence were determined by relative read depth. Dot color indicates the founder colony and phenotype group as indicated in the legend. (PDF 113Â kb)

Additional file 1: Fig. S1. Map of the positions of the 14 study villages and their relation to the Niger River. Generated by Google Maps Professional.

Additional file 2: Fig. S2. A schematic presentation of bait station with a magnification of the permeable, black plastic membrane penetrable enough for mosquitoes to feed. B) Photo of the bait station hung on the outer wall of a house.

Concordance of allele frequencies as called from pooled sequence and Sequenom individual genotyping. Concordance was assessed with data from three Sequenom plexes spread across all 3 candidate loci, 3.1, 9.1, and 9.2. SNPs were chosen independently of the phenotype group; that is, they were chosen on the basis of differing allele frequencies in either of the founder pools, and not for high allele differences between phenotype pools. Allele frequencies for Sequenom data were calculated from genotype calls and allele frequencies from pool sequence were determined by relative read depth. Dot color indicates the founder colony and phenotype group as indicated in the legend. (PDF 113Â kb)

Additional file 1: Fig. S1. Map of the positions of the 14 study villages and their relation to the Niger River. Generated by Google Maps Professional.

Verification of gene silencing for Toll10 and Toll 11. Four days after dsRNA injection (indicated above gel), cDNA synthesis was performed using M-MLV reverse transcriptase with random hexamers (Invitrogen). In each case, 1ug of total RNA was used in triplicate assays. Triplicates were pooled and used a template for PCR analysis of targeted (TOLL 10, TOLL 11) and control (ribosomal protein Rps7) genes (indicated on left margin of gel). (PDF 192Â kb)

Additional file 2: Fig. S2. A schematic presentation of bait station with a magnification of the permeable, black plastic membrane penetrable enough for mosquitoes to feed. B) Photo of the bait station hung on the outer wall of a house.

Verification of gene silencing for Toll10 and Toll 11. Four days after dsRNA injection (indicated above gel), cDNA synthesis was performed using M-MLV reverse transcriptase with random hexamers (Invitrogen). In each case, 1ug of total RNA was used in triplicate assays. Triplicates were pooled and used a template for PCR analysis of targeted (TOLL 10, TOLL 11) and control (ribosomal protein Rps7) genes (indicated on left margin of gel). (PDF 192Â kb)