Fig. 4: Development success and sex ratio of larvae with decreased support in bacterial metabolites.

In A–D, auxotrophic E. coli-colonized larvae were treated with 1× or 0.01× concentrations of D-Ala and m-DAP to slow bacterial development. A Number of colony forming units (CFU)/mL in wells containing individual larvae treated with 1× (black dots) or 0.01× (white dots) D-Ala and m-DAP. Each dot represents a well, 6 wells/time point/replicate were tested in at least 3 replicates. Bars represent the mean ± SEM. Time-points indicate the time after bacteria were added to germ-free larvae. B Proportion of mosquitoes reaching adulthood (dark grey), blocked in development (white) or dead (light grey) when auxotrophic E. coli-colonized larvae were supplemented with standard (1×) or diluted (0.01×) D-Ala and m-DAP concentrations. Bar charts represent the mean ± SEM of five independent replicates (individual points). C Proportion of males amongst the adults and D number of days until reaching adulthood when auxotrophic E. coli-colonized larvae were supplemented with standard (1×, black) or diluted (0.01×, white) D-Ala and m-DAP. Bars represent the mean ± SEM. In (E, F), larvae were colonised with wt E. coli, the growth deficient mutant ∆mnmA or the biotin-deficient ∆iscUA mutant. E Proportion of mosquitoes reaching adulthood (dark grey), blocked in development (white) or dead (light grey) when larvae were colonised with different E. coli mutants. Bar charts represent the mean ± SEM of five independent replicates (individual points). F Proportion of males amongst the adults when larvae were colonised with different E. coli mutants. Bars represent the mean ± SEM. Numbers below graphs indicate the number of wells/mosquitoes analysed per replicate and condition.