In the present study, we developed an automatic mass-injection system (AMIS) to produce an extract of infected H. illucens larvae (iHIL-E) and then evaluated antimicrobial peptide (AMP) expressions and assessed the antimicrobial activity of iHIL-E against various pathogens and Lactobacillus species. AMP gene expressions were assessed by real-time quantitative polymerase chain reaction (PCR) and the antimicrobial activities of iHIL-E were estimated using a radial diffusion assay and by determining minimal inhibitory concentrations. Results showed that the antimicrobial activity of HIL extract was effectively enhanced by L. casei infection and that the gene expressions of cecropin 3 and defensin 3 (antimicrobial peptides) were up-regulated. iHIL-E also prevented the growths of Enterococcus faecalis, Streptococcus mutans, and Candida vaginitis (MICs 200, 500, and 1000 µg/100 µL, respectively) and demonstrated high protease resistance. Moreover, the growths of methicillin-resistant Staphylococcus aureus, antibiotic-resistant Pseudomonas aeruginosa and AMP-resistant bacteria, Serratia marcescens, and Pseudomons tolaasii were significantly suppressed by iHIL-E. In addition, although iHIL completely cleared Salmonella species at concentrations of >200 µg/100 µL, Lactobacillus species were unaffected by iHIL at concentrations of <1000 µg/100 µL. The present investigation shows that the devised automatic mass injection system is effective for the mass production of the extract of infected HIL and that this extract is a novel, natural, protease-resistant, antibiotic candidate with broad-spectrum antibiotic activity.
Keywords: Hermetia illucens larvae; antimicrobial peptide; automatic mass injection system; preservatives.