# Use of Autochthonous Lactobacillusli to Increase the Safety of Zgougou
## 2 Results
### Antioxidant Activity of Zgougou  
Zgougou juice had higher ferric reducing antioxidant power (FRAP) than the seeds, regardless of the inoculation of autochthonous lactobacilli (Figure 2A). When scavenging activity against 2,2-DiPhenyl-1-Picrylhydrazyl (DPPH) was used for estimating antioxidant activity, the activity increased depending on the incubation time applied in the assay, reaching the highest value after 120 min (Figure 2B). Overall, antioxidant activity of the zgougou juice and seeds was ever lower than ascorbic acid and BHT, used as positive controls. Significant differences among the samples were found only after 20 min of incubation. In detail, juice from fermentation with lactobacilli (J LAB+) and seeds obtained with spontaneous fermentation (S LAB-) showed higher DDPH scavenging activity than the two other samples. Taken together, these results would indicate that J LAB+ had the highest antioxidant activity (FRAP and DPPH tests).  
\begin{table}
\begin{tabular}{c c c c c} \hline
**Target Bacterial Species** & **J LAB–** & **J LAB+** & **S LAB–** & **S LAB+** \\ \hline _Staphylococcus aureus_ & – & + & – & + \\ _Staphylococcus epidermidis_ & – & + & – & + \\ _Microoccus luteus_ & + & + & – & + \\ _Escherichia coli_ & + & + & + & + \\ _Listeria monocytogenes_ & – & ++ & + & + \\ _Enterococcus faecalis_ & – & ++ & + & + \\ _Salmonella Typhimuriumium_ & – & + & + & + \\ _Bacillus cereus_ & + & + & – & + \\ _Vibrio parahaemolyticus_ & – & ++ & + & + \\ _Aeromonas hydrophilia_ & – & + & – & – \\ \hline \end{tabular}
\end{table}
Table 5: Antibacterial activity8 of juice (J) and seeds (S) obtained after 24 h of fermentation (spontaneous, LAB–, or inoculated, LAB+, with autochthonous lactobacilli) of Aleppo pine’s seeds.  
Antioxidant activity of puddings prepared using juice of Aleppo pine's seeds increased depending on the concentration (100, 200, 300, 400 and 500 g/L) of ethanolic extract used. The puddings showed higher FRAP value than the control pudding, produced without addition of zgougou juice (Figure 3A). At a concentration of 500 g/L, this did not depend on the use of autochthonous lactobacilli. However, at a lower concentration (100 g/L), the pudding prepared using J LAB+ had higher (\(p<0.05\)) FRAP value than the pudding with J LAB- and the control pudding. Results of DPPH radical scavenging activity confirmed those obtained using the FRAP assay, except for the concentration 100 g/L (Figure 3B). At this concentration, both puddings prepared with fermented juice had higher antioxidant activity than the control pudding.  
Figure 3: Antioxidant activity of ethanolic extracts of pudding prepared using fermented (P LAB-, spontaneously, or, P LAB+, inoculated with autochthonous lactobacilli) zgougou juice, as assessed through determination of FRAP assay after 30 min (**A**) and DPPH radical scavenging activity after 120 min (**B**). Pudding P, produced without addition of fermented zgougou juice, was used as control. Ascorbic acid was used as positive control. BHT was used as additional positive control in the DPPH assay. The extract of each pudding was tested at 100 g/L (grey bars) or 500 g/L (white). Bars labelled with at least one common letter were not significantly different (\(p=0.05\)).  
Figure 2: Antioxidant activity of juice (J) and seeds (S) obtained after spontaneous fermentation (LAB-) or fermentation with autochthonous lactobacilli (LAB+), as assessed through determination of ferric reducing antioxidant power (FRAP) assay after 30 min (**A**) and 2,2-DiPhenyl-1-Picrylhydrazyl (DPPH) radical scavenging activity after 0 (black bars), 20 (grey) and 120 (light grey) min (**B**). Ascorbic acid was used as positive control in both the assays. Butyl hydroxy toluene (BHT) was used as additional positive control in the DPPH assay. Within readings performed at the same incubation time, bars labelled with at least one common letter were not significantly different (\(p=0.05\)).