Fig. 5

C.sp_P-produced adulticidal factor is non-volatile. Filtered (i.e. cell-free) C.sp_P biofilm supernatant and LB were spun in open containers in a vacuum centrifuge for 30 min at room temperature to allow for evaporation of volatile compounds. Original volume of vacuum centrifuged (VC) samples was then reconstituted using sterile water. Cell-free C.sp_P biofilm supernatant (VC and not) or LB as a control (VC and not) were mixed 1:1 with 3% sucrose (1.5% final sucrose concentration) and fed to An. gambiae females for 24 h. Experimental sugar meals were then removed and survival was monitored daily. The experiment was repeated four independent times, with 26–30 individuals per treatment per replicate. Survival analysis was performed using a Cox proportional hazards model testing the effect of bacterial treatment (C.sp_P vs LB), vacuum centrifugation, and an interaction between the two factors. No interaction was detected, indicating that the effect of C.sp_P ingestion was consistent regardless of whether samples were vacuum centrifuged. Main effects of both bacterial treatment (z = 9.15, P < 0.0001) and vacuum centrifugation (z = 4.08, P < 0.0001) were significant. In pairwise comparisons between their respective LB controls, C.sp_P (z = 5.95, P < 0.0001) and C.sp_P VC (z = 7.11, P < 0.0001) treatments had highly significant effects on survival