*2.2. Microbiota Analysis*

Fecal samples were shipped frozen to the George Washington University Genomics Core for processing. Each sample underwent DNA and RNA extraction in parallel from 250 mg of fecal material using a ZymoBIOMICS DNA/RNA Miniprep Kit (Zymo Research Corporation, Irvine, CA, USA). The resulting DNA was quality controlled using a Thermo Fisher Qubit 3.0 High Sensitivity DNA kit (Life Technologies, Carlsbad, CA, USA) and standardized to 0.2 ng/μL for library construction. Sequencing libraries were prepared, along with a ZymoBIOMICS Microbial Community DNA standard, using a Nextera XT DNA Library Prep kit (Illumina, San Diego, CA, USA) following Illumina's recommended guidelines. Libraries were quality controlled using a Thermo Fisher Qubit 3.0 High Sensitivity DNA kit and an Agilent 2100 Bioanalyzer High Sensitivity DNA kit (Agilent, Santa Clara, CA, USA) and were subsequently sequenced as paired-end, 2 × 150 bp, using a NextSeq 500 Mid-Output kit (Illumina, San Diego, CA, USA), with a 1% phi X sequencing control spike-in.
