# Evaluation of Non-Encapsulated and Microencapsulated Lactic Acid Bacteria
## 3 Results
### Molecular Identification of Lactic Acid Bacteria  
The LAB strains based on phenotypical characteristics were exposed to molecular confirmation. From the total of 15 isolates strains that were biochemically identified (API 50 CHL and Absi online), only 12 isolates were confirmed by 165 rRNA analysis when \begin{table}
\begin{tabular}{c c} \hline \hline
**Strains/Code IBNA** & **Area of Intestinal Content and Age of broiler Chickens** \\ \hline \multicolumn{2}{c}{_L. acidophilus_ biqutype 3, IBNA} \\
\begin{tabular}{c} Ileum, 26 d-old \\ Cecum, 26 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\ \hline _L. acidophilus_ biotype 1, IBNA 64 & Ileum, 45 d-old \\ \hline _L. brevis_ biotype 2, IBNA & \\
24 &
\begin{tabular}{c} Cecum, 26 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
50 &
\begin{tabular}{c} Ileum, 45 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\ \hline \multicolumn{2}{c}{_L. fermentum_ biotype 1, IBNA} \\
25 &
\begin{tabular}{c} Ileum, 26 d-old \\ Ileum, 26 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
56 &
\begin{tabular}{c} Ileum, 45 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
57 &
\begin{tabular}{c} Ileum, 45 d-old \\ \end{tabular} \\ \hline \multicolumn{2}{c}{_L. salivarius_, IBNA} \\
29 &
\begin{tabular}{c} Ileum, 45 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
33 &
\begin{tabular}{c} Ileum, 45 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
29 &
\begin{tabular}{c} Ileum, 45 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
33 &
\begin{tabular}{c} Ileum, 45 d-old \\ Ileum, 45 d-old \\ \end{tabular} \\
41 &
\begin{tabular}{c} Ileum, 45 d-old \\ \end{tabular} \\ \hline \hline \end{tabular}
\end{table} 
Table 1: The origin of _Lactobacillus_ strains existence in broiler chickens GIT. the primers Lac1F/ Lac1R were used as following: _L. acidophilus_ (IBNA 26, IBNA 64, 2/15 isolates; 13.33%), _L. fermentum_ (IBNA 37, 1/15 isolates; 6.66%), _L. salivarius_ (IBNA 29, IBNA 33, IBNA 41, 3/15 isolates; 20%), species belonging to _Lactobacillus_ genus (IBNA 24, IBNA 25, IBNA 56, IBNA 57, IBNA 48, IBNA 61, 6/15 isolates; 40%) (Table 2). Thus, the BLAST analysis used to achieve the molecular diagnostic of LAB strains included in this study revealed a 100% identification with similar 165 rRNA sequences from GenBank in the case of previously mentioned strains. However, for other strains, identification was possible only to the _Lactobacillus_ genus level, or did not confirm the characterization based on phenotypical features as was the case of two samples (IBNA 27 and IBNA 51) of _L. acidophilus_ that were classified as _L. johnsonii_ on a molecular basis. To eliminate any ambiguity, and for an accurate classification, the samples that were classified as _Lactobacillus_ spp. or as _L. johnsonii_ (instead of _L. acidophilus_ accordingly to API test) were sequenced again by using the pair of primers 27F/ 1492R that amplify the entire 165 rRNA gene. The new results, also classified _L. acidophilus_ biotype 3 IBNA, samples 27 and 51 as _L. johnsonii_. Instead, for the rest of the isolates the new BLAST analysis based on the complete 165 rRNA sequence confirmed the classification based on the API test (Table 2). Considering the 15 isolates identified from _Lactobacillus_ genus, the following rank for frequency of species identification was registered: _L. salivarius, L. acidophilus_, and _L. fermentum_. After molecular identification, compared with the biochemical results, the six _Lactobacillus_ isolates (IBNA 26, IBNA 64, IBNA 37, IBNA 29, IBNA 33, and IBNA 41) were identified and evaluated for desirable probiotic-related properties and their growth behavior fermentation process. The strains which were noted to have a significant resistance to low pH, bile salts, antibiotics susceptibility were exposed for further assay.