Escherichia coli (EC), Staphylococcus aureus (SA), Bacillus subtilis (BS), Rhodopseudomonas palustris (RP), Saccharomyces cerevisiae (SC) and Lactobacillus plantarum (LP) were selected as feed additives for black soldier fly (Hermetia illucens) by tracking the growth performance, proximate composition, digestive ability and antibacterial peptides (AMPs) content in the first trial. Microorganism efficiency screening results showed that RP could improve growth performance, digestive ability and AMP content of H. illucens. Therefore, RP was selected to prepare the diets and was incorporated into diets for H. illucens at levels of 0 (R0), 1.22 × 106 (R1), 1.22 × 107 (R2), 1.22 × 108 (R3), 1.22 × 109 (R4) and 1.22 × 1010 (R5) CFU/g. After 5 d of feeding, larvae fed the R2-R5 diets had higher weight gain and specific growth rates. Different concentrations of RP had no significant effect on larval body composition. R4–R5 could improve the digestibility and expression of AMPs in larvae. Moreover, RP could significantly increase the abundance of Lactobacillus and Rhodopseudomonas and decrease the abundance of Proteus and Corynebacterium. Therefore, RP is superior to the other strains as a feed additive for H. illucens larvae, and we recommend the addition of 1.22 × 109–1.22 × 1010 CFU/g RP to promote the growth and AMP content of H. illucens.