## 3 Results
### Microbiome Analysis  
#### 3.4.1 Taxonomy Assignment  
Proteobacteria (\(\mu\) = 86.909%) was found majorly before treatment, whereas the amount of Firmicutes (\(\mu\) = 36.012%) was increased after treatment in G1 samples. Bacteroidetes, Actinobacteria, Fusobacteria, Spirochaetes, TM7, Synergistetes, SR1, Tenericutes, GN02, Verrucomicrobia, and cyanobacteria phylum were also found in G1 samples (Figure 3a). Similarly, Proteobacteria (\(\mu\) = 81.711%) was found majorly before treatment, whereas the amount of Firmicutes (\(\mu\) = 41.440%) was increased after treatment in G2 samples. Bacteroidet, Actinobacteria, Fusobacteria, TM7, SR1, Spirochaetes, GN02, Synergistetes, Tenericutes, and Verrucomicrobia phylum were also found in G2 samples (Figure 3b). \begin{table}
\begin{tabular}{c c c c c} \hline \hline \multirow{2}{*}{**No.**} & \multirow{2}{*}{**Variables**} & \multicolumn{3}{c}{**Differences**} \\ \cline{3-5}  & & **Mouthwash** & **Mouthwash** & **_p-Value_** \\  & & **Formulation I (G1)** & **Formulation II (G2)** & \\ \hline
1. & Plaque index & \(-0.375\) & \(-0.376\) & 0.884 \\
2. & Gingival index & \(-0.071\) & \(-0.063\) & 0.923 \\
3. & Probing depth & \(-0.048\) & \(-0.093\) & 0.346 \\
4. & Bleeding on probing & \(-0.071\) & \(-0.063\) & 0.923 \\ \hline \hline \end{tabular}
\end{table} 
Table 6: The Mann–Whitney test compares the plaque index, probing depth, gingival index, and bleeding on probe between G1 and G2. \begin{table}
\begin{tabular}{c c c c c} \hline \hline \multirow{2}{*}{**Samples**} & \multirow{2}{*}{**Initial (Units/mL)**} & \multicolumn{2}{c}{**3 Months**} \\  & & **(Units/mL)** & **_p-Value_** \\ \hline Mouthwash formulation I stored at 4 \({}^{\circ}\)C & 359.13 \(\pm\) 6.67 & 353.62 \(\pm\) 6.27 & 0.5372 \\ Mouthwash formulation I stored at RT & 384.76 \(\pm\) 7.88 & 377.66 \(\pm\) 0.78 & 0.2254 \\ Mouthwash formulation I stored at 4 \({}^{\circ}\)C & 364.10 \(\pm\) 3.55 & 378.30 \(\pm\) 3.55 & 0.136 \\ Mouthwash formulation II stored 4 \({}^{\circ}\)C & 373.05 \(\pm\) 2.84 & 358.85 \(\pm\) 2.84 & 0.1580 \\ Mouthwash formulation II stored at RT & 361.83 \(\pm\) 0.00 & 353.45 \(\pm\) 3.83 & 0.2254 \\ Mouthwash formulation II stored at 45 \({}^{\circ}\)C & 366.59 \(\pm\) 3.34 & 365.88 \(\pm\) 4.05 & 0.2254 \\ \hline \hline \end{tabular}
\end{table} 
Table 4: The stability of mouthwash formulations I and II are based on enzyme activity. RT: room temperature. \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline \multirow{2}{*}{**No.**} & \multirow{2}{*}{**Variables**} & \multicolumn{3}{c}{**Mouthwash Formulation I (G1; \(n\) = 8)**} & \multicolumn{3}{c}{**Mouthwash Formulation II (G2; \(n\) = 10)**} \\ \cline{3-5}  & & **Pre-Treatment** & **Post-Treatment** & **(Mean \(\pm\) SE)** & **_p-Value_** \\  & & **(Mean \(\pm\) SE)** & **(Mean \(\pm\) SE)** & **(Mean \(\pm\) SE)** & **(Mean \(\pm\) SE)** & **_p-Value_** \\ \hline
1. & Plaque index & 2.33 \(\pm\) 0.13 & 1.95 \(\pm\) 0.14 & 0.0001 \({}^{\star}\) & 2.11 \(\pm\) 0.12 & 1.73 \(\pm\) 0.11 & 0.0001 \({}^{\star}\) \\
2. & Gingival index & 0.07 \(\pm\) 0.07 & 0.00 \(\pm\) 0.00 & 0.3173 & 0.06 \(\pm\) 0.06 & 0.00 \(\pm\) 0.00 & 0.3173 \\
3. & Probing depth & 2.18 \(\pm\) 0.12 & 2.13 \(\pm\) 0.11 & 0.3625 & 2.34 \(\pm\) 0.12 & 2.25 \(\pm\) 0.08 & 0.0143 \({}^{\star}\) \\
4. & Bleeding on probing & 0.07 \(\pm\) 0.07 & 0 & 0.3173 & 0.06 \(\pm\) 0.06 & 0.00 \(\pm\) 0.00 & 0.3173 \\ \hline \hline \end{tabular}
\end{table} 
Table 5: The comparison of the plaque index, probing depth, gingival index, and bleeding on probe values of pre-and post-treatment samples of G1 and G2. \({}^{\star}\) indicates the statistical significance. The OTUs can be divided into several categories according to their occurrences among different samples in groups before and after the mouthwash treatment. The relative frequency of the phyla can be used to identify the efficacy of the DMW.