The microbial taxonomy profiling was performed using Kraken2 against the microbial database

• Purpose: Identify potential pathogens from black soldier fly derived compost, which is an important value added product. • Time Period: June-Aug 2023 • Geographical Coverage: The actual samples were collected in Texas, USA. But the rearing methods and outputs occur globally. • Methodology: Samples of BSF derived organic material was obtained from a USA based company. Genomic DNA was extracted according to the manufacturer’s protocol using the Qiagen DNeasy Plant Pro Kit. Resulting genomic DNA was used for NGS library preparation and sequencing. Libraries were prepared using the Roche Kapa HyperPrep DNA library kit with Kapa Unique Dual-Indexed adapters following manufacturer’s recommendations. Completed libraries were QC’d and quantified using a combination of Qubit dsDNA HS and Agilent 4200 TapeStation HS DNA1000 assays. The libraries were normalized to a consistent concentration and equal volumes of the normalized libraries pooled, and the pool was quantified using the Invitrogen Collibri Quantification qPCR kit. This pool was loaded onto one lane of a NovaSeq SP flow cell using the Xp loading kit & workflow. Sequencing was performed in a 2x150bp paired end format using a NovaSeq 6000 v1.5 300 cycle reagent kit. Base calling was done by Illumina Real Time Analysis (RTA) v1.18.54 and output of RTA was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v2.20.0. Trimmomatic v0.39 was used to remove adaptor sequences and the reads were mapped to the human genome using Bowtie2 v2.5.1 for contaminates. The microbial taxonomy profiling was performed using Kraken2 against the microbial database. Pavian (https://fbreitwieser.shinyapps.io/pavian/) was used to visualize the metagenomics classification results.