# Use of Autochthonous Lactobacillusli to Increase the Safety of Zgougou
## 2 Results
### Cultivable Microbiota of Juice and Seeds from Fermented Zgougou  
Autochthonous _L. paraplantarum_ A1 and _L. plantarum_ A2, used in combination as starters, acidified the mixture of Aleppo pine's seeds and water from an initial pH of ca. 5.85 to a final pH (after 24 h of fermentation) of ca. 4.38. Acidification was observed also for the spontaneously, non-inoculated fermented zgougou, although at a lesser (\(p<0.05\)) extent. Indeed, the final pH reached the value of ca. 4.65.  
After 1 h of fermentation, presumptive coccus-shaped and rod-shaped LAB dominated the cultivable bacterial biota of non-inoculated juice (J LAB-) and seeds (S LAB-) (Figure 1, panels A and B). At the end of fermentation of the non-inoculated juice, presumptive mesophilic cocci were found at highest cell density, followed by coliforms and the other remaining groups of presumptive LAB. During fermentation, micrococci and staphylococci increased their number of ca. 5 log cycles, but remained sub-dominant compared to LAB and coliforms. High cell densities (8-9 log CFU/g) of presumptive mesophilic cocci and coliforms were found in the non-inoculated seeds after 24 h of fermentation. Other LAB groups, micrococci and staphylococci were found at sub-dominant levels. During fermentation of non-inoculated zygough, enterococci increased their number of 3-4 log cycles, reaching a final cell density in the order of 5 log CFU/mL (juice) or g (seeds).  
\begin{table}
\begin{tabular}{c c} \hline
**Microorganism** & **pH** \\ \hline _Lactobacillus parapplantarum_ A1 & 4.65 \(\pm\) 0.02\({}^{\mathrm{f}}\) \\ _Lactobacillus plantarum_ A2 & 4.65 \(\pm\) 0.01\({}^{\mathrm{f}}\) \\ _L. plantarum_ A3 & 4.75 \(\pm\) 0.03\({}^{\mathrm{e}}\) \\ _Enterococcus faecalis_ A4 & 4.84 \(\pm\) 0.02\({}^{\mathrm{e}}\) \\ _Ent. faecalis_ A5 & 4.79 \(\pm\) 0.02\({}^{\mathrm{de}}\) \\ _Ent. faecalis_ A6 & 4.89 \(\pm\) 0.01\({}^{\mathrm{b}}\) \\ _Ent. faecalis_ A7 & 4.96 \(\pm\) 0.01\({}^{\mathrm{a}}\) \\ _Ent. faecalis_ A8 & 4.83 \(\pm\) 0.02\({}^{\mathrm{c}}\) \\ _Ent. faecalis_ A9 & 4.85 \(\pm\) 0.03\({}^{\mathrm{c}}\) \\ _Ent. faecalis_ A10 & 4.76 \(\pm\) 0.02\({}^{\mathrm{e}}\) \\ _Ent. faecalis_ A11 & 4.92 \(\pm\) 0.03\({}^{\mathrm{ab}}\) \\ _Ent. faecalis_ A12 & 4.80 \(\pm\) 0.01\({}^{\mathrm{d}}\) \\ _Ent. faecalis_ A13 & 4.85 \(\pm\) 0.02\({}^{\mathrm{c}}\) \\ _Ent. faecalis_ A14 & 4.93 \(\pm\) 0.02\({}^{\mathrm{a}}\) \\ \hline \end{tabular}
\end{table}
Table 1: pH values of zgougou inoculated with autochthonous lactic acid bacteria after 24 h of fermentation at 25 \({}^{\circ}\)C.  
Figure 1: Cell densities (log CFU/mL or log CFU/g) of different bacterial groups found after 1 (white bars) and 24 (black bars) h of zgougou fermentation. (**A**) juice obtained after spontaneous fermentation (J LAB-); (**B**) seeds obtained after spontaneous fermentation (S LAB-); (**C**) juice obtained after fermentation with autochthonous lactobacilli (J LAB+); (**D**) seeds obtained after fermentation with autochthonous lactobacilli (S LAB+).  
After 1 h of fermentation of zgougou obtained through chemical sterilization of seeds and with addition of autochthonous lactobacilli (J LAB+ and S LAB+), presumptive mesophilic lactobacilli were found at the highest cell density (8.5 log CFU/mL or g), followed by thermophilic lactobacilli (Figure 1, panels C and D). The other bacterial groups, excepted total mesophilic microorganisms, were found at values lower than 4 log CFU/mL or g. After 24 h of fermentation, lactobacilli stayed dominant, being found at numbers higher than 9 log CFU/mL or g. Presumptive coccus-shaped LAB and coliforms increased their number, especially in the seeds. On the contrary, cell density of enterococci decreased below the limit of detection after 24 h of fermentation.