Considerable data by many experimental studies have provided that the S. persica plant and its chewing sticks (sewak) display beneficial effects for oral hygiene. To find out more about the shadowy benefits of sewak, we hypothesized that sewak can be a natural therapeutic approach that controls the microbiota composition. Surveys and mouthwash samples with and without the use of sewak were obtained from six healthy randomly selected volunteers. Samples were processed and DNA isolated using QlAamp kit. 16S metagenomics sequencing library approach (Illumina) for sequencing the variable V3 and V4 domains of the 16S rRNA gene was used since purposes such as genus and species were needed in this study. Each samples were sequenced to a depth of 380,000 reads in the paired-end 2x300 read format. Taxonomic identification and statistical analysis revealed that after using the sewak treatment, some species decreased and other increased (P \u3c 0.05). However, when the p-value was adjusted using FDR, results indicated that no one of the bacteria species can be considered as statistically significant (FDR\u3e0.05). The comprehensive examination of the relationship of sewak and the oral microbiota demonstrates that there is no difference in the entire number of bacteria before and after treatment, based on analysis. It is possible that sewak use may reduce oral bacteria in a global, proportional manner, such that the microbiome approach could not detect such changes, since the method is designed to measure composition changes, not absolute changes