Transformation of food waste to source of antimicrobial proteins by black soldier fly larvae for defense against marine Vibrio parahaemolyticus

Abstract

Insect biorefinery by black soldier fly larvae (BSFL), Hermetia illucens, has emerged as an innovative technique for the valorization of food waste. However, despite BSFL being an attractive natural source of antimicrobial proteins (AMPs), there is a scarcity of research on the antimicrobial activity and transcriptome expression of AMPs derived from BSFL following waste treatment. In the present study, food waste treatment was performed by BSFL with a substrate C/N ratio ranging from 21:1 to 10:1, marine Vibrio parahaemolyticus (VP) was selected as the model aquaculture pathogen, the antimicrobial activities of AMPs in vitro and zebrafish in vivo were examined, and the molecular mechanism of the C/N-dependent AMP difference was expounded. Findings were made that the AMP extract of C/N16:1 resulted in relatively higher antimicrobial activity in vitro than that of other C/Ns. Further, the AMPs of C/N16:1 exhibited a promising in vivo defense effect for elevating the 96-h survival rate of zebrafish from 0% to 39% after VP infection, comparable to the animal antibiotic sulfamethoxidine. The results of transcriptome analysis reveal that lysozymes were the highest expressed components in the AMP gene family. The C/N16:1 BSFL significantly up-regulated 12 out of 51 lysozyme genes compared with C/N21:1, which likely contributed to the improvement of AMP antimicrobial activity. Further, C/N16:1 significantly up-regulated the expression of lysozyme, glycosyl hydrolase and muscle protein genes compared with C/N21:1, which likely enhanced the defense ability of the immune system, the utilization of the starch-like substrate, and the mobility of the larvae, thereby facilitating the larval transformation and AMP production. Overall, such results indicate that waste C/N ratio interacted with the activity and expression of BSFL AMPs through transcriptome regulation, and the BSFL AMPs derived from food waste could be used for the defense against marine pathogens to support the sustainable development of aquaculture.

Keywords: Antimicrobial proteins; Black soldier Fly larvae; C/N ratio; Food waste; Transcriptome; Vibrio Parahaemolyticus.