Black Soldier Fly Gut Microbiota Resists Invasion by Bacillus subtilis 168 and Pseudomonas putida KT2440

Due to its high bioconversion efficiency and nutritional value, the black soldier fly (Hermetia illucens L. 1758) is a promising insect species for sustainable animal feed production. However, concerns remain regarding microbial safety when larvae are reared on substrates contaminated by pathogenic or spoilage bacteria. This study investigated the effects of substrate inoculation with Bacillus subtilis 168 or Pseudomonas putida KT2440 on larval performance and gut microbiota composition. Larvae reared on contaminated diets showed no significant differences in survival or development time compared to controls. However, a short-term reduction in growth was observed in the Bacillus-exposed group. qPCR analyses confirmed the temporary presence of Bacillus taxa in larval guts, while Pseudomonas taxa were effectively excluded. Amplicon sequencing of the 16S rRNA gene revealed that the contamination did not affect gut bacterial microbiota richness and composition. Instead, the bacterial communities evolved naturally with Lactobacillales-related bacteria dominating early stages and Morganellaceae taxa becoming more abundant in prepupae. Our findings demonstrate the stability and resilience of H. illucens gut bacterial microbiota, reinforcing the safety and suitability of H. illucens as a feed ingredient, even when reared under challenging microbial conditions.