Microorganisms inhabiting fermented foods represent the main link between the consumption of this food and human health. Although some fermented food is a reservoir of potentially probiotic microorganisms, several foods are still unexplored. This study aimed at characterizing the probiotic potential of lactic acid bacteria isolated from zgougou, a fermented matrix consisting of a watery mixture of Aleppo pine′s seeds. In vitro methods were used to characterize the safety, survival ability in typical conditions of the gastrointestinal tract, and adherence capacity to surfaces, antimicrobial, and antioxidant activities. Strains belonged to the Lactobacillus plantarum group and Enterococcus faecalis showed no DNase, hemolytic, and gelatinase activities. In addition, their susceptibility to most of the tested antibiotics, satisfied some of the safety prerequisites for their potential use as probiotics. All the strains tolerated low pH, gastrointestinal enzymes, and bile salts. They displayed a good antibacterial activity and antibiofilm formation against 10 reference bacterial pathogens, especially when used as a cell-free supernatant. Furthermore, the lactic acid bacteria (LAB) strains inhibited the growth of Aspergillus flavus and Aspergillus carbonarius. Finally, they had good antioxidant activity, although depending on the strain. Overall, the results of this work highlight that zgougou represents an important reservoir of potentially probiotic LAB. Obviously, future studies should be addressed to confirm the health benefits of the LAB strains.

Seeds of Pinus halepensis are used for preparing zgougou, a spontaneously fermented matrix giving juice and seeds debris, consumed in many Arabian countries, including Tunisia. In the same way as all the food processes based on spontaneous fermentation, zgougou hides health risks due to eventual pathogenic microorganisms and derived toxins. This study aimed at investigating the effect of the use of autochthonous Lactobacillus paraplantarum A1 and Lactobacillus plantarum A2, as fermentation starters, on the microbiological characteristics, profiles of volatile organic compounds (VOC), antibacterial and antioxidant activities of juice and seeds debris from zgougou. The starter lactobacilli inhibited undesired bacteria (e.g., Enterobacter and Aeromonas) and coccus-shaped lactic acid bacteria, as shown by culture-dependent and-independent methods. The inhibitory effect was more evident in juice than in seeds debris. Some VOC (ethanol, acetoin, phenol,2-methoxy and caryophyllene) were present at higher concentrations in juice and seeds obtained upon spontaneous fermentation, compared to the samples deriving from fermentation with lactobacilli. The latter samples were characterized by higher concentrations of acetic acid, decane, 1-nonanol, bornyl acetate and bornyl formate. In addition, they showed a wider spectrum of antibacterial activity than spontaneously fermented juice and seeds. The use of autochthonous lactobacilli did not relevantly affect the antioxidant activity of zgougou. When juice from lactobacilli-driven fermentation was used to prepare a traditional Tunisian pudding ("Assidat-Zgougou"), it improved color and odor with respect to the pudding containing juice from spontaneous fermentation. This study showed that the use, at laboratory scale, of autochthonous lactobacilli is a feasible biotechnological tool to outgrow undesired bacteria, thus improving the safety of zgougou juice. Future studies should be undertaken to confirm the observed benefits at industrial scale.