Gametocytemia and COI for <i>P</i>. <i>falciparum</i> field isolates.

Gametocytemia and COI for P. falciparum field isolates.</p

Additional information on <i>P</i>. <i>falciparum</i> isolates.

MSP1-genotyping results for P. falciparum isolates (p1-p6) used in infections with An. gambiae (An. gam), An. coluzzii (An. col), and/or VK5-collected An. coluzzii (VK5-An. col) (See also Methods). (XLSX)</p

Full-factor GLMM Output for VK5 <i>An</i>. <i>coluzzii</i>.

Effect of egg development (eggs vs no eggs), oocyst number, and their interactions on oocyst size. In this model, egg development and oocyst number were considered as fixed effects whereas parasite isolate, mosquito generation, and mosquito ID were set as random effects. Significant effects are in bold. (XLSX)</p

Full-factor GLMM Output for <i>An</i>. <i>coluzzii</i>.

Effect of treatment (dsGFP vs dsEcR), oocyst number, egg number, and their interactions on oocyst size. In this model, treatment, oocyst number and egg number were considered as fixed effects whereas parasite isolate and mosquito ID were set as random effects. Significant effects are in bold. (XLSX)</p

Egg development for individual infections.

(A-B) The effects of EcR-silencing on egg development across individual infections for (A) An. coluzzii (unpaired t-test and Mann-Whitney) and (B) An. gambiae (unpaired t-test and Mann-Whitney) compared to controls (Cntrl). N = sample size. p# = parasite isolate. (TIF)</p

Full-factor GLMM Output for <i>An</i>. <i>gambiae</i>.

Effect of treatment (dsGFP vs dsEcR), oocyst number, egg number, and their interactions on oocyst size. In this model, treatment, oocyst number and egg number were considered as fixed effects whereas parasite isolate and mosquito ID were set as random effects. Significant effects are in bold. (XLSX)</p

<i>An</i>. <i>coluzzii</i> and <i>An</i>. <i>gambiae</i> females were infected with <i>P</i>. <i>falciparum</i> field isolates following <i>EcR</i>-silencing.

(A-B)An. coluzzii and An. gambiae females were injected with either dsGFP (Cntrl) or dsEcR and then infected with P. falciparum field isolates originating from six different gametocyte carriers (designated patient, p#). (A) Two batches of An. coluzzii females were infected with parasites from p1, p2, and p3. (B) Three batches of An. gambiae females were infected with parasites from p1, p3, p4, p5, and p6. (C) In An. coluzzii, dsEcR-injections did not reduce cumulative egg development to the level of statistical significance (Mann-Whitney). (D) In An. gambiae, EcR-silencing did result in a significant reduction in cumulative egg numbers compared to controls (unpaired t-test). (E-F) In both species, (E) An. coluzzii and (F) An. gambiae, dsEcR-injections had no effect on cumulative oocyst prevalence (Fisher’s Exact) or intensity (unpaired t-test and Mann-Whitney). P next to pie charts = prevalence. N = sample size. p# = parasite isolate.</p

Oocyst prevalence and intensity for individual infections.

(A-B) The effects of EcR-silencing on the prevalence (Fisher’s Exact) and intensity (unpaired t-test and Mann-Whitney) of oocysts across individual infections for (A) An. coluzzii and (B) An. gambiae, compared to dsGFP-injected (Cntrl) females. P = oocyst prevalence. N = sample size. p# = parasite isolate. (TIF)</p

Sporozoite prevalence and intensity for individual infections.

(A) The salivary glands of dsEcR and dsGFP (Cntrl) An. gambiae females were assessed at 12 days post-infected blood meal from infections with parasite isolates p1, p3, p5, and p6. The sporozoite prevalence (Fisher’s Exact) and intensity (Mann-Whitney) for each infection are shown. P = sporozoite prevalence. N = sample size. p# = parasite isolate. (TIF)</p

Individual <i>P</i>. <i>falciparum</i> isolates vary in their growth response to <i>EcR</i>-silencing.

(A) For An. coluzzii females, both p1 and p2 parasites produced larger oocysts in EcR-silenced females, but p3 parasites failed to respond to dsEcR conditions and did not grow larger than controls (GLMM, LRT). (B) For An. gambiae, all P. falciparum isolates responded to EcR-silencing by growing larger except for p6 parasites, which remained the same size as controls (Cntrl), even though they were fed to the same batch of mosquitoes as p5 parasites (GLMM, LRT). (A-B) Oocyst size does not vary significantly between parasite isolates in dsGFP controls for An. gambiae or An. coluzzii (An. coluzzii dsGFP: GLMM, LRT X22 = 1.52, p = 0.466; An. gambiae dsGFP: GLMM, LRT X24 = 3.47, p = 0.483), yet it does for dsEcR-treated females in both species (An. coluzzii dsEcR: GLMM, LRT X21 = 6.3, p = 0.012; An. gambiae dsEcR: GLMM, LRT X21 = 4.67, p = 0.031). (C) Oocyst size varies significantly between An. coluzzii and An. gambiae control females infected with the same parasite isolates (p1 and p3) (GLMM, LRT), whereby oocysts are larger in An. coluzzii than in An. gambiae. (D) An. coluzzii were collected as larvae from VK5, Burkina Faso and infected with P. falciparum isolates p1-p6. Pre-gravid females that failed to develop eggs were less likely to become infected (pie charts, P = oocyst prevalence, Fisher’s Exact), although among infected individuals, there was no difference in oocyst intensity (Mann-Whitney). (E) Pre-gravid females had significantly larger oocysts at 7 days pIBM compared to gravid females (GLMM, LRT). For all applicable panels, average oocyst size per midgut is shown for simplicity–analyses are based on nested data incorporating all oocyst measurements. N = sample size, or number of mosquitoes. Noocysts = number of individual oocyst measurements. p# = parasite isolate.</p