Influence of Catecholamines (Epinephrine/Norepinephrine) on Biofilm Formation and Adhesion in Pathogenic and Probiotic Strains of Enterococcus faecalis.

Enterococcus faecalis has controversial status due to its emerging role in nosocomial infections, while some strains with beneficial effects are used as probiotics and starter cultures in dairy industry. These bacteria can be found as resident or transient germs in the gut or on skin, where they are continually exposed to various eukaryotic molecules. In this context, the aim of our work was to evaluate the effect of the catecholamine stress hormones, epinephrine (Epi), and norepinephrine (NE) on some Enterococcus strains. Four E. faecalis strains were included in this study: E. faecalis MMH594 and E. faecalis V583, pathogenic strains of clinical origin, E. faecalis Symbioflor 1 clone DSM 16431, a pharmaceutical probiotic, and E. faecalis OB15, a probiotic strain previously isolated from Tunisian rigouta (Baccouri et al., 2019). Epi was found to modulate the formation of biofilm (biovolume and thickness) in E. faecalis, whether pathogens or probiotics. NE had less effect on biofilm formation of these bacteria. We also investigated the effect of Epi and NE on adhesion of E. faecalis to eukaryotic cells as it is the first step of colonization of the host. Epi was found to significantly enhance the adhesion of MMH594 and OB15 to Caco-2/TC7 intestinal cells and HaCaT keratinocyte cells, whereas NE significantly increased the adhesion of V583 and Symbioflor 1 DSM 16431 to Caco-2/TC7 cells, the adhesion of MMH594, Symbioflor 1 DSM 16431, and OB15 to HaCaT cells. Analysis of a putative adrenergic sensor of Epi/NE in E. faecalis, compared to QseC, the Escherichia coli adrenergic receptor, allowed the identification of VicK as the nearest protein to QseC with 29% identity and 46% similarity values. Structure modeling and molecular docking of VicK corroborated the hypothesis of possible interactions of this putative adrenergic sensor with Epi and NE, with binding energies of -4.08 and -4.49 kcal/mol, respectively. In conclusion, this study showed for the first time that stress hormones could increase biofilm formation and adhesion to eukaryotic cells in E. faecalis. Future experiments will aim to confirm by in vivo studies the role of VicK as adrenergic sensor in E. faecalis probiotic and pathogen strains. This may help to develop new strategies of antagonism/competition in the gut or skin ecological niches, and to prevent the colonization by opportunistic pathogens.

Draft Genome Sequence of Enterococcus faecalis Strain OB15, a Probiotic Strain Recently Isolated from Tunisian Rigouta Cheese.

Enterococcus faecalis OB15 is a probiotic strain that was isolated from rigouta, a popular traditional Tunisian fermented cheese. We report here the draft genome sequence of this strain, consisting of 2,912,159 bp, with an average G+C content of 37.49%.

Probiotic Potential and Safety Evaluation of Enterococcus faecalis OB14 and OB15, Isolated From Traditional Tunisian Testouri Cheese and Rigouta, Using Physiological and Genomic Analysis.

Lactic acid bacteria (LAB) strains OB14 and OB15 were isolated from traditional Tunisian fermented dairy products, Testouri cheese and Rigouta, respectively. They were identified as Enterococcus faecalis by the MALDI TOF-MS (matrix assisted laser desorption-ionization time of flight mass spectrometry) biotyper system and molecular assays (species-specific PCR). These new isolates were evaluated for probiotic properties, compared to E. faecalis Symbioflor 1 clone DSM 16431, as reference. The bacteria were found to be tolerant to the harsh conditions of the gastrointestinal tract (acidity and bile salt). They were low to moderate biofilm producers, can adhere to Caco-2/TC7 intestinal cells and strengthen the intestinal barrier through the increase of the transepithelial electrical resistance (TER). Susceptibility to ampicillin, vancomycin, gentamicin and erythromycin has been tested using the broth microdilutions method. The results demonstrated that E. faecalis OB14 and OB15 were sensitive to the clinically important ampicillin (MIC = 1 µg/mL) and vancomycin (MIC = 2 µg/mL) antibiotics. However, Whole Genome Sequencing (WGS) showed the presence of tetracycline resistance and cytolysin genes in E. faecalis OB14, and this led to high mortality of Galleria Mellonella larvae in the virulence test. Hierarchical cluster analysis by MALDI TOF-MS biotyper showed that E. faecalis OB15 was closely related to the E. faecalis Symbioflor 1 probiotic strain than to OB14, and this has been confirmed by WGS using the average nucleotide identity (ANI) and Genome-to-Genome Hybridization similarity methods. According to these results, E. faecalis OB15 seems to be reliable for future development as probiotic, in food or feed industry.

Comparative study on volatile compounds from Tunisian and Sicilian monovarietal virgin olive oils.

The effects of ripening degree of olives on volatile profile of monovarietal virgin olive oils (VOO) from Tunisian and Sicilian cultivars were investigated. Fruits obtained from Tunisia (Chétoui and Chemlali) and Italy (Nocellara del Belice, Biancolilla and Cerasuola) were picked at three different stages of ripeness and then immediately processed. Moreover, the changes in volatile composition were evaluated in Chétoui variety as a function of the irrigation regime versus the rain-fed control. Using headspace-solid-phase microextraction (HS-SPME) technique coupled to GC-MS and GC-FID, the volatile compounds of the monovarietal virgin olive oils were identified and quantitatively analyzed. The proportions of different classes of volatiles of oils showed significant differences throughout the maturity process. The results suggest that adding to the genetic factor; agronomic conditions affect the volatile formation and therefore the organoleptic properties of VOO.

Chemical composition and oxidative stability of Tunisian monovarietal virgin olive oils with regard to fruit ripening.

The chemical composition of virgin olive oil may be influenced by genotype and different agronomic (i.e. fruit ripeness degree, water supply) and technological factors. This article reports the evaluation of the influence of the olive ripening stage on the quality indices, the major and the minor components and the oxidative stability of the two main monovarietal Tunisian cultivars (cvv. Chétoui and Chemlali) virgin olive oils. Moreover, the olives cv. Chétoui were tested in a rain-fed control and an irrigation regime. The oils sampled at five different ripeness stages were submitted to liquid chromatographic determination (HPLC-DAD/MSD) of their quali-quantitative phenolic and tocopherolic profiles. Moreover, the triacylglycerol and fatty acid compositions, and minor components such as squalene, pigments and their relation with the oil oxidative stability were evaluated. The tested oils showed very good correlation between the oxidative stability and the concentrations of total phenols, practically secoiridoids and α-tocopherol.