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The 16S rRNA sequencing process included polymerase chain reaction (PCR) amplification: Primers were designed for the V4 region of the 16S rRNA gene, with the primer sequences being 515F and 806R

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Fluorescence signals were visualized using a fluorescence microscope (Nikon Ts2R) and quantified by flow cytometry (NovoCyte 3000, Agilent, USA)