Relative contributions of egg-associated and substrate-associated microorganisms to Black Soldier Fly larval performance and microbiota

Larvae of the black soldier fly (BSF) can be used to convert organic waste into insect biomass for animal feed. In this process, they interact with microorganisms originating from the substrate, the insect, and the environment. The substrate is the main determinant of the larval gut microbiota composition, but inoculation of the substrate with egg-associated bacteria can improve larval performance. We aimed to quantify the relative importance of substrate-associated and egg-associated microorganisms on BSF larval performance, bacterial abundance, and microbiota composition, when larvae were fed with chicken feed or chicken manure. For this we inactivated substrate-associated microorganisms by autoclaving, or disinfected BSF eggs. Larval survival rate, weight, and proportion of prepupae were determined on day 15. We collected substrate and larval samples on days 0 and 15 and performed 16S rRNA gene targeted qPCR and amplicon sequencing. In both chicken feed and chicken manure, egg disinfection did not cause any difference in larval performance or overall microbiota composition. In contrast, in chicken manure, substrate-associated microorganisms increased larval biomass and autoclaving caused major shifts in microbiota. Thus, substrate-associated microorganisms not only impact the larval microbiota but also larval performance, whereas egg-associated microorganisms have a minor role in the densities present