Amplification and sequencing of bacterial 16S rRNA was performed according to our previous reports [25]. The V3-V4 region of the 16S rRNA gene of bacteria was amplified by polymerase chain reaction (PCR) using the following primers: forward, 5′-TCGTCGGCAGCGTCAGATGTGTATAAGCGACAGCCTACGGGNGGCWGCAG-3′, and reverse, 5′-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC-3′, which was followed by the addition of the sequencing adapters. The amplicons were sequenced using the Illumina MiSeq platform (Illumina Inc., San Diego, CA, USA). A total of 10,000 reads per sample were randomly selected for further analysis. Samples with insufficient read numbers were re-sequenced, and those with repeated insufficient read numbers were excluded.
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