## 3 Results  
The NMCC-11 strain was classified as a probiotic as a result of in vitro morphological, biochemical, and probiotic studies (Figure 1). Strain NMCC-11 showed a 100% homology with _Pediococcus acidilactici_, according to NCBI results (Table 1). The NMCC-11 isolate was confirmed as a Gram-positive coccus using sample microscopy (Figure 1), and after, negative catalase and oxidase tests were established (Table 1). For additional microbiological assessment, the viability of strain under various conditions was tested. The pure isolate was grown on MRS agar medium at 37 \({}^{\circ}\)C for 24 h, which resulted in circular, white, and smooth-textured colonies (Figure 1). The identification of presumed probiotic bacterial strains by molecular means is a critical step in gaining access to and assessing the safety of microorganisms of animal origin. The isolate was provisionally identified as _Pediococcus acidilactici_ after molecular identification employing the 165 rRNA gene (Table 1). The phylogenetic tree in Figure 2 shows the evolutionary history of _Pediococcus acidilactici_. The phylogenetic tree was constructed using MEGA-X from the 16S rRNA gene sequence using the neighbor-joining algorithm and shows the closely related species. The types of strains used for comparison are given along with their accession numbers (Figure 2). The NMCC-11 strain was grown at different temperatures on MRS agar and NaCl concentration in M-17 broth for 24-48 h. The amyloidytic, proteolytic, and cellulolytic activity were tested along with bacteriocin production (Table 2). The NMCC-11 strain demonstrated optimum growth at temperatures 37 \({}^{\circ}\)C and 0.5% of NaCl; however, the viability of this probiotic bacterial strain was estimated at different temperatures and concentrations of NaCl (Table 2). Positive results were also achieved for amyloidytic, proteolytic, and cellulolytic activity and bacteriocin production. The NMCC-11 strain also demonstrated tolerance to a wide pH range of 4 to 6 (Figure 3). \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Colony Morphological Characterization** & **Identification Based on 16S rRNA Sequence** \\ \hline
**Gram staining** & **Positive cocci** & **Strain ID** & NMCC-11 \\ \hline
**Shape** & Circular & & \\ \hline
**Surface** & Smooth & & \\ \hline
**Size** & \textgreater{}0.098 mm & & \\ \hline
**Color** & White & & \\ \hline
**Margin** & Entire & & \\ \hline
**Elevation** & Raised & & \\ \hline
**Biochemical characterization** & & & \\ \hline
**Catalase** & Negative & **Closely related published** & _Pediococcus acidilactici_ \\ \hline
**Oxidase** & Negative & & \\ \hline
**Glucose fermentation** & Positive & **Sequence similarity** & 99.21\% \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Biochemical and morphological characterization of _Pediococcus acidilactici_ and its identification based on the 16S rRNA gene sequence, along with the published accession numbers in the DNA database. Figure 2: Schematic representation of the phylogenetic tree showing closely related species interfered from the 16S rRNA gene. No significant differences, in terms of bacterial viability, among different pH levels were observed at Time 0 (Figure 3A). Conversely, after 2 h (Time 1), a significant increase in bacterial viability (from 8.45 \(\pm\) 0.04 log CFU/mL to 9.23 \(\pm\) 0.31 and 9.35 \(\pm\) 0.1 log CFU/mL, **_p_ \(<\) 0.01) was observed at pH 5 and 6 compared to pH 4, respectively. A similar trend was observed after 4 h (Time 2); a significant increase in bacterial viability (from 8.70 \(\pm\) 0.05 log CFU/mL to 9.65 \(\pm\) 0.05 and 9.72 \(\pm\) 0.07 log CFU/mL, ***_p_ \(<\) 0.0001) was observed at pH 5 and 6 compared to pH 4, respectively. By producing essential organic acids, hydrogen peroxide, and certain antimicrobial substances and bacteriocins, NMCC-11 also showed a good antimicrobial effect against _Pseudomonas aeruginosa_ (ATCC 9027), _Staphylococcus aureus_ (ATCC 6538), _Listeria monocytogenes_ (ATCC 13932), and _Bacillus cereus_ (ATCC 11778) (Table 3).