Many studies reported the positive synergy between polyphenols and probiotics. Probiotics are known for their positive and beneficial effects on human intestinal health, and their interactions with polyphenolic compounds and metabolites may increase health benefits improving intestinal barrier function and modulating the gut microbiota. Here a symbiotic strategy of quercetin (QUE) and Lactobacillus acidophilus LA5 antimicrobial and antiadhesion properties against E. coli was evaluated on intestinal Caco-2 differentiated cells. Our study demonstrates that QUE improves LA5 auto-aggregation and LA5 and E. coli coaggregation. Additionally, LA5 decreased the adhesion of E. coli to Caco-2 cells mainly via exclusion antagonism than competition assay. Interestingly, QUE did not affect LA5 effects on E. coli adhesion, but synergically improved the barrier function of Caco-2 cells improving TEER values, claudin-1 expression and reducing proinflammatory COX-2 expression. Data confirmed the LA5 and QUE synergism supporting their combined use in the prevention or treatment of pathogen diseases.

Endophthalmitis most likely originates from both planktonic bacteria suspended in the tear film and bacteria adherent to the conjunctiva and the eyelid. This study aimed to expand the research on the effectiveness of a colloidal silver solution (Silverix®) against ocular microorganisms. The activity of Silverix® was evaluated against methicillin-resistant Staphylococcus aureus, S. epidermidis, ofloxacin-resistant Pseudomonas aeruginosa, and Candida albicans strains, previously characterized for their antibiotic resistance and biofilm-forming capabilities. The microbial killing was estimated at various times in the presence and absence of colloidal silver solution against planktonic and biofilm-embedded cells. The results documented the efficacy of Silverix® on planktonic cells of S. aureus and S. epidermidis (2.49–2.87 Log CFU/mL reduction) and P. aeruginosa strains (3–4.35 Log CFU/mL reduction). On the contrary, C. albicans showed mild susceptibility. Regarding early biofilm, the ocular isolates were harder to kill (2–2.6 Log CFU/mL reduction) than the reference strains, whereas a similar decrease (3.1 Log CFU/mL reduction) was estimated for P. aeruginosa strains. The light microscope images of biofilms treated with colloidal solution confirmed the ability of Silverix® to destroy the biofilm.

The synthesis of contaminant-free silver@linear carbon chains (Ag@LCCs) nanohybrid systems, at different Ag/LCCs ratios, by pulsed laser ablation was studied. The ablation products were first characterized by several diagnostic techniques: conventional UV-Vis optical absorption and micro-Raman spectroscopies, as well as scanning electron microscopy, operating in transmission mode. The experimental evidence was confirmed by the theoretical simulations' data. Furthermore, to gain a deeper insight into the factors influencing metal@LCCs biological responses in relation to their physical properties, in this work, we investigated the bioproperties of the Ag@LCCs nanosystems towards a wound-healing activity. We found that Ag@LCC nanohybrids maintain good antibacterial properties and possess a better capability, in comparison with Ag NPs, of interacting with mammalian cells, allowing us to hypothesize that mainly the Ag@LCCs 3:1 might be suitable for topical application in wound healing, independent of (or in addition to) the antibacterial effect.

Saponaria sicula Raf. grows in Sicily, Sardinia, and Algeria on limestone cliffs and volcanic sands 1300–2500 m above sea level. The aim of the present study was to investigate how the pedo-climatic conditions influence the micromorphological, phytochemical, and biological properties of Sicilian S. sicula leaves collected in the Madonie Mountains (SsM) and on Etna Mt (SsE). Micromorphological investigations revealed that leaves from SsM had a higher amount of calcium oxalate druses in the mesophyll and a more intense blue–green staining with Toluidine blue O, indicating a higher content of polyphenols. These data were confirmed by phytochemical analyses carried out on hydroalcoholic extracts, which showed a higher content of total phenols (8.56 ± 0.57 g GAE/100 g DE) and flavonoids (6.09 ± 0.17 g RE/100 g DE) in SsM. Sixty-four compounds were identified by LC-DAD-ESI-MS analysis with propelargonidin dimer as the most abundant compound (10.49% and 10.19% in SsM and SsE, respectively). The higher polyphenol content of SsM leaves matches also with their biological activity, identifying SsM extract as the strongest plant complex (IC50 2.75–477.30 µg/mL). In conclusion, the present study experimentally demonstrates that not only climatic differences but also soil characteristics affect the micromorphological, phytochemical, and biological features of this plant species.

Pistachio (Pistacia vera L.) nuts contain nutrients and phytochemicals which have been linked to several positive outcomes. The aim of this research was to examine the antimicrobial effect of natural raw and roasted unsalted polyphenols-rich pistachio extracts (NRRE and RURE) and hexane oil fractions. American Type Culture Collection (ATCC), food and clinical isolates of Gram-positive bacteria (Listeria monocytogenes and Staphylococcus aureus), Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli and Enterococcus faecium) and yeasts (Candida albicans) were used. In addition, the influence of the extraction method was evaluated. Generally, NRRE extracts were richer in polyphenolic compounds compared with RURE extracts. NRRE extracted with n-hexane was the most effective on Listeria monocytogenes food isolates strains (MIC values between 0.25 and 2.0 mg mL−1). All extracts, except for RURE extracted with n-hexane, were active against Listeria monocytogenes ATCC 13932. Both hexane oil fractions were active against Listeria monocytogenes ATCC 13932 and Enterococcus faecium DSZM 17050. The oil obtained from natural pistachio was active against three food isolates of Listeria monocytogenes. In conclusion, the present study indicates an inhibitory effect of pistachio polyphenols against Listeria monocytogenes, one of the most serious pathogens causing foodborne disease.

Punica granatum is a rich source of bioactive compounds which exhibit various biological effects. In this study, pomegranate peel and leaf ethanolic crude extracts (PPE and PLE, respectively) were phytochemically characterized and screened for antioxidant, antimicrobial and antiviral activity. LC-PDA-ESI-MS analysis led to the identification of different compounds, including ellagitannins, flavonoids and phenolic acids. The low IC50 values, obtained by DPPH and FRAP assays, showed a noticeable antioxidant effect of PPE and PLE comparable to the reference standards. Both crude extracts and their main compounds (gallic acid, ellagic acid and punicalagin) were not toxic on Vero cells and exhibited a remarkable inhibitory effect on herpes simplex type 1 (HSV-1) viral plaques formation. Specifically, PPE inhibited HSV-1 adsorption to the cell surface more than PLE. Indeed, the viral DNA accumulation, the transcription of viral genes and the expression of viral proteins were significantly affected by PPE treatment. Amongst the compounds, punicalagin, which is abundant in PPE crude extract, inhibited HSV-1 replication, reducing viral DNA and transcripts accumulation, as well as proteins of all three phases of the viral replication cascade. In contrast, no antibacterial activity was detected. In conclusion, our findings indicate that Punica granatum peel and leaf extracts, especially punicalagin, could be a promising therapeutic candidate against HSV-1.

Studies carried out using three different in vitro assays and a biological setting (Escherichia coil) demonstrated the antioxidant activity of Scutellaria lateriflora microshoot extract. Moreover, the extract exhibited no toxicity in a brine shrimp lethality bioassay. These results indicated that microshoots are a rich, safe source of antioxidants, which encouraged us to enhance their production in vitro. In agar and agitated cultures, two biotechnological strategies were applied: feeding the cultures with the biogenetic precursors of the phenolics—phenylalanine and tyrosine, and eliciting them with methyl jasmonate. Specific Scutellaria flavonoids and verbascoside were analysed by HPLC. Feeding with precursors (1 g/L) in agar cultures decreased the production of the metabolites. In agitated cultures, different concentrations of precursors (1.0–2.5 g/L) and the elicitor (10; 50; 100 µM) were tested. Additionally, parallel feeding with the precursor and elicitor in a concentration of 50 µM were applied. The best strategy for total flavonoid and verbascoside production was phenylalanine feeding (1.5 g/L), max. 3765 and 475 mg/100 g DW, respectively, after 7 days. This is the first report documenting the high antioxidant production in S. lateriflora microshoots after feeding with phenylalanine. Moreover, for the first time, bioreactor cultures were successfully maintained, obtaining attractive results (max. total flavonoid content 2348 and verbascoside 485 mg/100 g DW).

Owing to the richness of bioactive compounds, Olea europea leaf extracts exhibit a range of health effects. The present research evaluated the antibacterial and antiviral effect of leaf extracts obtained from Olea europea L. var. sativa (OESA) and Olea europea var. sylvestris (OESY) from Tunisia. LC-DAD-ESI-MS analysis allowed the identification of different compounds that contributed to the observed biological properties. Both OESA and OESY were active against Gram-positive bacteria (MIC values between 7.81 and 15.61 μg/mL and between 15.61 and 31.25 μg/mL against Staphylococcus aureus ATCC 6538 for OESY and OESA, respectively). The antiviral activity against the herpes simplex type 1 (HSV-1) was assessed on Vero cells. The results of cell viability indicated that Olea europea leaf extracts were not toxic to cultured Vero cells. The half maximal cytotoxic concentration (CC50) values for OESA and OESY were 0.2 mg/mL and 0.82 mg/mL, respectively. Furthermore, both a plaque reduction assay and viral entry assay were used to demonstrate the antiviral activity. In conclusion, Olea europea leaf extracts demonstrated a bacteriostatic effect, as well as remarkable antiviral activity, which could provide an alternative treatment against resistant strains.

Brassica incana Ten. is an edible plant belonging to the Brassicaceae family. In this work, the phenolic composition and the antioxidant and cytotoxic properties of the hydroalcoholic extracts obtained from the leaves and the flowering tops of B. incana grown wild in Sicily (Italy) were studied for the first time. A total of 17 and 20 polyphenolic compounds were identified in the leaf and in the flowering top extracts, respectively, by HPLC-PDA-ESI-MS analysis. Brassica incana extracts showed in vitro antioxidant properties; the leaf extract displayed greater radical scavenging activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test than the flowering top extract (IC50 = 1.306 ± 0.049 mg/mL and 2.077 ± 0.011 mg/mL), which in turn had a stronger ferrous ion chelating ability than the other (IC50 = 0.232 ± 0.002 mg/mL and 1.147 ± 0.016 mg/mL). The cytotoxicity of the extracts against human colorectal adenocarcinoma (CaCo-2) and breast cancer (MCF-7) cell lines was evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the lactic dehydrogenase (LDH) release determination. The extracts showed cytotoxic efficacy against Caco-2 cells, with the flowering top extract being the most effective (about 90% activity at the highest concentration tested). In the brine shrimp lethality bioassay, the extracts exhibited no toxicity, indicating their potential safety.

Aim: To develop electrospun mats loaded with Thymus capitatus (L.) essential oil ( ThymEO) and to study their morpho-mechanical and antimicrobial properties. Materials & methods: Poly(lactic acid) (PLA) mats containing ThymEO were prepared by electrospinning. The effect of ThymEO on the morpho-mechanical properties of fibers was assayed by scanning electron microscopy and dynamometer measurements. The antimicrobial activity of ThymEO delivered either in liquid or vapor phase was assessed through killing curves and invert Petri dishes method. The cytotoxicity was also investigated. Results: The mechanical properties were enhanced by integrating ThymEO into PLA. Both liquid and vapors of ThymEO released from mats caused reductions of microbial viable cells. Negligible cytotoxicity was demonstrated. Conclusion: PLA/ ThymEO delivery systems could be suitable for treating microbial infections.

Background Candida sp. represent the most common cause of fungal infections worldwide. In the present work, we have evaluated the activity of an essential oil extracted from pistachio hulls against a number of standard and clinical strains of Candida sp. Methods C. albicans ATCC 64550, C. parapsilosis ATCC 22019, 4 clinical strains of C. albicans , 3 clinical strains of C. parapsilosis and 3 clinical strains of C. glabrata were used. All clinical isolates were identified by species-specific PCR-based methods. Susceptibility studies were performed using pistachio hull essential oil alone or in combination with antifungal compounds. The interactions between pistachio hull essential oil and selected antifungal compounds were also evaluated using the checkerboard method and the mechanisms of interaction investigated by droplet size distribution. Results Pistachio hull essential oil was fungicidal at the concentrations between 2.50 and 5.0 mg/ml. D-limonene and 3-Carene were the components with major activity. An antagonistic effect was observed with all combinations tested. Conclusion The antifungal activity of pistachio hull essential oil could be used to help control resistance in Candida species. More studies need to be performed to elucidate the mechanisms responsible for the activity of pistachio hull essential oil.

Bacteria colonization on the lid margin could play a crucial role in developing of biofilm causes of chronic infections such as blepharo-conjunctivitis or lacrimal system disease. The aim of this study was to evaluate the efficacy of an ophthalmic fixed antibiotics/steroid combination (AC) containing tetracycline (TET), chloramphenicol (CAF) and colistin (CS), in the ratio 2:1:3, on bacterial planktonic growth and preformed biofilm. The antimicrobial effect of AC against Staphylococcus aureus 815 and Pseudomonas aeruginosa 1 ocular strains was evaluated by time-killing curves analysis at 30’, 2, 4, 6, 8 and 24 hr. The activity of AC (exposure of 2 and 6 hr) on preformed 24 hr-biofilm was evaluated by cell viability and biomass measurements, expressed as logarithm of colony-forming units/ml (Log 10 CFU/ml) and optical density values, respectively. Time-killing curves showed that AC was able to reduce the load of planktonic growth of both strains versus control. A significant reduction of 5 Log 10 CFU/ml after 8h for S. aureus and after 4h for P. aeruginosa was reported. The AC was active against preformed biofilm, with a viability significant reduction (>2 Log 10 CFU/ml versus control) of both strains at the exposure times. A biomass reduction of 40% and 60% for S. aureus and P. aeruginosa respectively, was detected at the exposure time of 2 hr and maintained at 6 hr. The three antibiotics CAF, TET and CS present in AC showed a significant antimicrobial and antibiofilm effect. Antibiotics combination is a promising strategy for increasing treatment efficacy and for controlling resistance evolution. The results of this study, taken together, suggest the applications of this fixed combination in the treatment of chronic ocular infections where planktonic cells and biofilm are involved. antibiotics combination, biofilm, ocular infection, lacrimal system

Ficus vasta Forssk. (Moraceae) is traditionally used for the treatment of various ailments; nonetheless, this species has been poorly studied to date. This work aimed to characterize the phenolic profile and to evaluate the antioxidant and antimicrobial properties of a hydroalcoholic extract obtained from F. vasta leaves collected in Egypt.

Carvacrol (CAR) is one of the most promising essential oil components with antimicrobial activity. New technologies aimed to incorporate this active molecule into carrier matrix to improve the stability and prolong the biological activity. The goal of this study was to investigate the feasibility of incorporating CAR into electrospun membranes of poly(lactic acid) (PLA) for potential applications as active antimicrobial system. To this end, PLA membranes containing homogeneously dispersed CAR were successfully prepared and a series of systematic tests including morpho-mechanical properties, in vitro release rate, and antimicrobial/antibiofilm activities against Staphylococcus aureus and Candida albicans were carried out. The results revealed that CAR has a good compatibility with PLA and acts as a plasticizer, improving flexibility and extensibility of the matrix. The gradual release of CAR from PLA membranes warranted a significant antimicrobial activity up to 144 h and reduced the biofilm production by 92–96 and 88–95% of S. aureus and C. albicans in single and mixed cultures. A strong decrease of cell count, biomass, metabolic activity, and vitality of established 24- and 48-h biofilms were also demonstrated. In conclusion, this work highlights the potential of electrospun nanofibrous membranes as efficient stabilizers-carriers of CAR and opens up interesting perspectives on the use of this system as new tool for skin and wound bacterial–fungal infections.

PurposeTo evaluate the antifungal activity of a fixed antibiotic combination (AC) containing tetracycline (TET), chloramphenicol (CAF), and colistimethate sodium (CS).MethodsIn vitro: Candida ATCC and clinical strains were used. The minimum inhibitory concentrations (MICs) of AC and of each antibiotic were determined. Fluconazole (FLC) was tested for comparison. Time-killing curves of selected strains were performed. Ex vivo keratitis: corneas were injected intrastromally with the selected strains. After the injection, corneas were divided into groups of treatments: AC, FLC, or saline. Then, the tissues were analyzed for colony-forming units per gram (CFU/g). Propidium iodide (PI) and MitoTracker (MTR) staining were used to investigate the mode of action.ResultsValues of MIC required to inhibit the growth of 90% of organisms for the antibiotics alone were higher than FLC. However, their activity was enhanced when used in combination against Candida yeasts. Time-killing curves showed that at 24 hours, AC reduced the load of both strains of approximately 1 Log10 CFU/g compared with the initial inoculum (P < 0.0001). This effect was also significant versus FLC. In ex vivo, AC was effective in decreasing the loads of both strains by 4 Log10 CFU/g with respect to the control. Moreover, it showed higher activity than FLC against Candida albicans ATCC 10231 (1 Log10 CFU/g, P < 0.01 versus control). PI staining demonstrated that CS changed the membrane's permeability, whereas MTR staining demonstrated that TET or CAF altered mitochondrial function. The cells treated with AC and stained showed both effects.ConclusionsIn this study, AC showed antifungal efficacy versus Candida spp.; this activity can be due to the synergistic effects of antibiotics in it.

The role of nitric oxide (NO) as an antimicrobial and anticancer agent continues to stimulate the search of compounds generating NO in a controlled fashion. Photochemical generators of NO are particularly appealing due to the accurate spatiotemporal control that light-triggering offers. This contribution reports a novel molecular construct in which multiple units of 3-(trifluoromethyl)-4-nitrobenzenamine NO photodonor are clustered and spatially organized by covalent linkage to a calix[4]arene scaffold bearing two quaternary ammonium groups at the lower rim. This multivalent calix[4]arene-NO donor conjugate is soluble in hydro-alcoholic solvent where it forms nanoaggregates able to release NO under the exclusive control of visible light inputs. The light-stimulated antibacterial activity of the nanoconstruct is demonstrated by the effective bacterial load reduction of Gram-positive Staphylococcus aureus ATCC 6538 and Gram-negative Escherichia coli ATCC 10536.

Essential oils from the leaves and fruits of Litsea cubeba Pers. collected in the Assam and Arunachal Pradesh states in north-east India, were analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). On the whole, 117 components have been characterized. The two leaf oils (LC1, LC2) show sabinene as the main component; the other significant compounds for LC1 oil are -pinene, terpinen-4-ol, -terpineol and myrcene, whereas for LC2 1,8-cineole and -pinene are the other most important compounds. The three fruit oils (LC3, LC4 and LC5) were characterized by different profiles, indeed LC3 and LC4 showed a similar composition with citronellol and citronellal the main components, accounting for 70% and 10% of total oils, respectively. LC5, instead, presents geranial (c. 44%) and neral (c. 40%) as the main components, whereas citronellal reaches only c. 3%. Essential oils were evaluated for their antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. All microbial strains appeared sensitive to the cytotoxic activity of the essential oils under investigation. Leaf and fruit oils showed different levels of inhibition depending on their particular chemical composition; however, the LC5 sample was broadly the most effective.

Staphylococcal growth and biofilm formation in culture medium where pH was lowered with weak organic (acetic and lactic) or strong inorganic (hydrochloric) acids were studied. The effects were evaluated by biomass measurements, cell-surface hydrophobicity, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). The results demonstrated that the inhibition was related to type of acidulant and pH value. At pH 5.0, the antibacterial effect was more pronounced in the presence of acetic acid (58-60% growth reduction) compared with that in the presence of lactic (7-16% growth reduction) and hydrochloric acids (23-24% reduction). The biofilm biomass of Staphylococcus aureus and Staphylococcus epidermidis was reduced by 92, 85, 63, and 93, 87, 81% after exposition to acetic, lactic, and hydrochloric acids, respectively. Increasing the pH from 5.0 to 6.0 resulted in a noticeable reduction in the effectiveness of acids. A minor cells hydrophobic character was also documented. The SEM and CLSM revealed a poorly structured and thinner biofilm compared with the dense and multilayered control. Acidic environment could have important implications for food-processing system to prevent bacterial colonization and control biofilm formation. The findings of this study lead to consider the rational use of the type of acid to achieve acidic environments.

Biofilms are a serious problem, cause of severe inconvenience in the biomedical, food and industrial environment. Staphylococcus aureus and S. epidermidis are important pathogenic bacteria able to form thick and resistant biofilms on various surfaces. Therefore, strategies aimed at preventing or at least interfering with the initial adhesion and subsequent biofilm formation are a considerable achievement. The aim of this study was to evaluate the effect of alkaline pH on bacterial adhesion and further biofilm formation of S. aureus and S. epidermidis strains by biofilm biomass, cell-surface hydrophobicity, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) analysis. The results demonstrated that the amount of biofilm biomass formed and the surface hydrophobicity were significantly less than what were observed at higher levels of pH. SEM and CLSM images revealed a poorly structured and very thin biofilm (2.53 times thinner than that of the controls). The inhibiting effect of the alkaline pH on the bacterial attachment impaired the normal development of biofilm that arrested at the microcolony stage. Alkaline formulations could be promising towards the control of bacterial colonization and therefore the reduction of the biofilm-related hazard. In the clinical setting, alkaline solutions or cleaners could be promising to prevent the bacterial colonization, by treating surfaces such as catheters or indwelling medical devices, reducing the risk of biofilm related infections.