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.

Despite herbal medicine being popular across the Mediterranean basin, there is no evidence in favor of COVID-19 infection. This study investigates the utilization and effects of medicinal plants in Italy, Lebanon, and Tunisia during COVID-19 and its effects on post-COVID-19 pandemics. We used a tailored, web-based “Google Form” questionnaire with the random sampling method. We gathered 812 complete responses (Italy: 116, Lebanon: 557, and Tunisia: 139), revealing diverse demographics and symptom experiences. Fatigue prevailed across all groups (89.0–94.2%), while psychological impacts ranged from 20.1% to 30.9%, with higher rates in Lebanon. Post-COVID-19 symptoms affected 22.4% (Italy), 48.8% (Lebanon), and 31.7% (Tunisia). General use of herbs was consistent (41.4–50.4%), with 23.3% (Italy), 50.2% (Lebanon), and 65.5% (Tunisia) employing herbs for COVID-19 therapy. Notably, in Lebanon, Za’atar, a thyme-like plant, correlated with reduced symptoms, suggesting potential protective effects that are likely due to its polyphenol richness. This study underscores the persistent reliance on traditional medicinal plants remedies in the Mediterranean area, with regional variations. Further exploration of herbal compounds for COVID-19-like symptoms is warranted.

The intestinal barrier is a complex and dynamic structure that plays a critical role in maintaining gut health. Though, pathological conditions such as, dysbiosis or inflammation that could damage the intestinal barrier to varying degrees, aggravating the primary disease. Recently, fermented plant-origin foods and probiotics were used as an alternative for health remedy. For this reason, in this current study, we made the inquiries about the effect of Tunisian zgougou juice obtained upon both spontaneous (J) and driven fermentation (J(A1+A2)) with autochthonous probiotic bacteria Lactiplantibacillus plantarum (LP) species A1 and A2, the whole bacteria (A1) and (A2) and their CFS. The cell integrity and permeability was assessed based on the TEER measurement on monolayer cell culture of Caco-2 cell, then the gene expression levels of the tight junction proteins (TJP) and the gene expression level of the inflammatory cytokine TNFα were evaluated using the RT-PCR. Our experimental findings exhibited that the driven fermented had a potential protective effects on intestinal barrier function in vitro shown by the great increase the electrical barrier detection (TEER) between Caco2 cell reaching ca. 690 to 963.33 Ω/cm2, although the regulatory mechanism has not been reported yet. Otherwise LP (A1 and A2) and their CFS showed moderate results. Furthermore, J(A1+A2) promoted the mRNA relative gene expression of the different tested tight TJPs and moderated the intestinal inflammation by influencing the TNFα expression. This study revealed that probiotics and especially probiotic-based plant-origin fermented functional foods (Tunisian zgougou juice) could improve and reinforce the intestinal barrier which provide new ideas for treating intestinal injury-related diseases, and have to be a revival in worldwide popularity, mostly as a consequence of claims made about their health benefits. So that, to completely reveal the health advantages of these probiotic LAB strains for humans and this new food matrix, additional in vivo and clinical investigations are necessary, as highlighted by the limitations of in vitro models.

The intestinal barrier is a critical regulator of gut health, often compromised by dysbiosis and inflammation. Given the rising interest in fermented plant‐based foods and probiotics as adjuvants in maintaining gut health, this study explored the biological activity of zgougou juice, comparing spontaneous fermentation (J) with driven fermentation using two lactobacilli with probiotic traits (namely Lactiplantibacillus paraplantarum A1 and Lactiplantibacillus plantarum A2), as well as the effects of cell biomass and Cell‐Free Supernatants (CFS) obtained from those lactobacilli. Using a Caco‐2 cell monolayer model, we assessed barrier integrity via Transepithelial Electrical Resistance (TEER) and gene expression of Tight Junction Proteins (TJP) and the cytokine Tumor Necrosis Factor α (TNFα). Treatment of Caco‐2 monolayer with zgougou juice fermented with lactobacilli significantly reinforced the intestinal barrier, with TEER values reaching 690–963.33 Ω·cm ² . Furthermore, it upregulated TJP expression, outperforming bacterial biomass and their CFS. These findings would suggest that zgougou juice improves intestinal barrier, though further in vivo and clinical trials are necessary to claim any benefits for human health.