Metagenomic sequencing of H. illucens larvae at Day 14(Se20 treatment group)
At each sampling interval (days 7 and 14), three biological replicates per group (CK and Se20) were collected for metagenomic analysis, with each replicate consisting of ten pooled larvae. Before dissection, larvae were starved for 12 h to evacuate gut contents, followed by surface sterilization via triple rinsing with 70% ethanol and sterile water. The gut tissues were then excised under aseptic conditions for immediate DNA extraction and metagenomic sequencing. Genomic DNA of the intestinal microbiota was extracted using TIANamp Soil DNA Kit (DP336, TIANGEN Biotech, Beijing, China). Subsequently, the concentration and quality of the resultant DNA were quantified spectrophotometrically (NanoDrop ND-1000, Thermo Fisher Scientific, Waltham, MA, USA). Shotgun metagenomic sequencing was conducted on an DNBSEQ-T7 platform with PE150 strategy (Wekemo Tech Group Co., Ltd., Shenzhen, China).
SUB16325686
MSe14-3