DNA was extracted from samples using the MN NucleoSpin 96 Soil kit, followed by quantification and purity assessment with a Nano Drop 2000 spectrophotometer. PCR amplification was performed on the V3-V4 region of the bacterial 16S rRNA gene using specific primers. The amplified products were purified and processed with Trimmatic and UCHIME software for quality filtering and chimera removal. OTU clustering was conducted at a 97% similarity threshold, and species annotation was performed using the RDA Classifier with a confidence threshold of 0.8. Functional predictions of microbial communities were made using PICRUSt software, analyzing each OTU for KO, Pathway, and EC information.