## 3 Results and Discussion
### Antimicrobial Activity
#### 3.5.2 Determination of the Minimum Inhibitory Concentration (MIC) of _P. rhoeas_ Extracts  
The MIC results of the _P. rhoeas_ samples were determined against most of the microbes studied. The MIC results of three samples and antibiotics are presented in Table 2. The MIC results of our samples against the studied bacteria range from 0.78 mg/mL for sample P3E against _E. coli_ ATCC and _E. coli_ CIP and 50 mg/mL for sample P2E against the same strains, while the MIC values of P1E and P2E samples against yeast _C. albicans_ are 50 and 25 mg/mL, respectively. Our values of MIC are also consistent with the literature when compared to the MIC values from previous research. A study in Turkey, the ethanolic extract of _P. rhoeas_ showed antibacterial effect against _S. aureus_ at 0.150 mg/mL but no action against _C. albicans_, _E. coli_, and _K. pneumoniae_. In addition, the MIC of diethyl ether, acetone, and chloroform extracts against _S. aureus_ was 39.06 g/mL [27]. In Morocco, the MICs of flower _P. rhoeas_ extract against _S. aureus_, _K. pneumoniae_, and _E. coli_ were 60, 30, and 60 mg/mL, respectively [39]. In another Moroccan research, the antimicrobial effect of the four organs (flower, leaf, stem, and root) of _P. rhoeas_ was determined. The MIC ranged from 0.78 mg/mL to 50 mg/mL [33]. In this regard, study [53] suggested that the presence of hydroxyl groups in phenolic substances may be related to their increased antimicrobial activity [54, 55]. [PERSON] et al., on the other hand, discovered a link between the content of alkaloids (particularly roemerine) in _P. rhoas_ extracts and their antibacterial potential [20]. In this context of the effect of the geographic region on plant metabolites and biological activities, recent studies confirm our results. A Chinese study on _P. ostii_ stamen extracts collected from eight different locations in China was conducted to demonstrate \begin{table}
\begin{tabular}{c c c c c c} \hline \multirow{2}{*}{**Sample**} & \multicolumn{3}{c}{**Gram – Bacteria**} & **Gram + Bacteria** & **Yeast** \\ \cline{2-5}  & _Escherichia coli_ & _Escherichia coli_ & _Klebsiella_ & _Staphylococcus aureus_ & _Candida albicans_ \\ \hline P1E & 12.66 \(\pm\) 0.57 b & 15.00 \(\pm\) 1.00 b & 13.33 \(\pm\) 1.52 ab & 13.66 \(\pm\) 0.57 b & R \\ \hline P2E & 13.00 \(\pm\) 0.00 b & 12.33 \(\pm\) 0.57 c & 10.66 \(\pm\) 1.15 b & 14.00 \(\pm\) 1.00 b & 10.00 \(\pm\) 2.00 b \\ \hline P3E & 13.00 \(\pm\) 0.00 b & 11.66 \(\pm\) 0.57 c & R & 12.33 \(\pm\) 0.57 b & R \\ \hline Tetracycline & 19.00 \(\pm\) 1.00 a & 18.10 \(\pm\) 0.70 a & 14.90 \(\pm\) 0.60 a & 16.70 \(\pm\) 0.90 a & — \\ \hline Streptomycin & R & R & R & 09.61 \(\pm\) 0.200 c & — \\ \hline Fluconazole & — & — & — & 21.20 \(\pm\) 04.20 a & \\ \hline \end{tabular}  
* R—resistant; “——— antibiotic is not compatible with this strain; a, b, and c—DIZ values of extracts with a significant difference against the same strain (\(p<\) 0.05).  
\end{table} 
Table 1: Diameter of inhibition zones of P1E, P2E, P3E, tetracycline, streptomycin, and fluconazole in millimeters. \begin{table}
\begin{tabular}{c c c c c c} \hline \multirow{2}{*}{**Sample**} & \multicolumn{3}{c}{**Gram – Bacteria**} & **Gram + Bacteria** & **Yeast** \\ \cline{2-5}  & _Escherichia coli_ & _Escherichia coli_ & _Klebsiella_ & _Staphylococcus aureus_ & _Candida albicans_ \\ \hline P1E & 25 & 25 & 25 & 25 & 50 \\ \hline P2E & 50 & 50 & 25 & 25 & 25 \\ \hline P3E & 0.78 & 0.78 & ND & 50 & ND \\ \hline Streptomycin & 0.25 & 0.50 & 0.003 & 0.062 & — \\ \hline Tetracycline & 0.25 & 0.25 & 0.062 & 0.003 & — \\ \hline Fluconazole & — & — & — & — & 0.40 \\ \hline \end{tabular}  
* ND—none detected; R—resistant; “——— antibiotic does not match this strain.  
\end{table} 
Table 2: Minimal inhibitory concentration of _P. rhoas_ extracts and antibiotics in mg/mL. the distinction of bioactive molecules and biological capacities between _P. otsii_ stamen extracts from different geographical locations [28].