Antimicrobial efficacy of Punica granatum Lythraceae peel extract against pathogens belonging to the ESKAPE group.

The improper use and abuse of antibiotics have led to an increase in multidrug-resistant (MDR) bacteria resulting in a failure of standard antibiotic therapies. To date, this phenomenon represents a leading public health threat of the 21st century which requires alternative strategies to fight infections such as the identification of new molecules active against MDR strains. In the last 20 years, natural extracts with biological activities attracted scientific interest. Following the One Health Approach, natural by-products represent a sustainable and promising alternative solution. Consistently, the aim of the present study was to evaluate the antimicrobial activity of hydro-alcoholic pomegranate peel extract (PPE) against MDR microorganisms belonging to Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. "ESKAPE" group pathogens. Through semiquantitative and quantitative methods, the PPE showed effective antimicrobial activity against Gram-positive and Gram-negative MDR bacteria. The kinetics of bactericidal action of PPE highlighted that microbial death was achieved in a time- and dose-dependent manner. High concentrations of PPE exhibited antioxidant activity, providing a protective effect on cellular systems and red blood cell membranes. Finally, we report, for the first time, a significant intracellular antibacterial property of PPE as highlighted by its bactericidal action against the staphylococcal reference strain and its bacteriostatic effect against clinical resistant strain in the HeLa cell line. In conclusion, due to its characterized content of polyphenolic compounds and antioxidant activity strength, the PPE could be considered as a therapeutic agent alone or in conjunction with standard antibiotics against challenging infections caused by ESKAPE pathogens.

Eco-Friendly Sanitization of Indoor Environments: Effectiveness of Thyme Essential Oil in Controlling Bioaerosol Levels and Disinfecting Surfaces.

Bioaerosols and pathogens in indoor workplaces and residential environments are the primary culprits of several infections. Techniques for sanitizing air and surfaces typically involve the use of UV rays or chemical sanitizers, which may release chemical residues harmful to human health. Essential oils, natural substances derived from plants, which exhibit broad antimicrobial properties, could be a viable alternative for air and surface sanitation. The objective of this study has been to investigate the efficacy of thyme essential oil (TEO) in environmental sanitation processes. In Vitro assays through agar well diffusion, disk volatilization and tube dilution methods revealed significant antimicrobial activity of TEO 100% against foodborne and environmental isolates, with both bacteriostatic/fungistatic and bactericidal/fungicidal effects. Therefore, aqueous solutions of TEO 2.5% and 5% were formulated for air sanitation through nebulization and surface disinfection via direct contact. Bioaerosol samples and surface swabs were analyzed before and after sanitation, demonstrating the efficacy of aqueous solutions of TEO in reducing mesophilic and psychrophilic bacteria and environmental fungi levels in both air and on surfaces. The obtained results prove the antimicrobial potential of aqueous solutions of TEO in improving indoor air quality and surface cleanliness, suggesting thyme essential oil as an effective and safe natural sanitizer with minimal environmental impact compared to dangerous chemical disinfectants.

Vine-Winery Byproducts as Precious Resource of Natural Antimicrobials: In Vitro Antibacterial and Antibiofilm Activity of Grape Pomace Extracts against Foodborne Pathogens.

Grape pomace is the main by-product of vine-winery chains. It requires adequate treatment and disposal but is also an economically underused source of bioactive plant secondary metabolites. This study aimed to investigate the antibacterial effects of polyphenolic extracts from Aglianico (Vitis vinifera L.) grape pomace. In particular, hydroethanolic extracts obtained via an ultrasonic-assisted extraction technique were selected for antimicrobial tests. The extracts were screened for their antibacterial effects against foodborne pathogens that were both Gram-positive, in the case of Staphylococcus aureus and Bacillus cereus, and Gram-negative, in the case of Escherichia coli and Salmonella enterica subsp. enterica serovar Typhimurium, showing variable bacteriostatic and bactericidal effects. In addition, our results demonstrated that the tested grape pomace extracts can reduce the inhibitory concentration of standard antibiotics. Interestingly, selected extracts inhibited biofilm development by S. aureus and B. cereus. Overall, these new insights into the antibacterial properties of grape pomace extracts may represent a relevant step in the design of novel therapeutic tools to tackle foodborne diseases, and in the management of resistant biofilm-related infections.

Antibacterial and Antibiofilm Efficacy of Thyme (Thymus vulgaris L.) Essential Oil against Foodborne Illness Pathogens, Salmonella enterica subsp. enterica Serovar Typhimurium and Bacillus cereus.

Nowadays, the wide spread of foodborne illness and the growing concerns about the use of synthetic food additives have shifted the focus of researchers towards essential oils (EOs) as possible antimicrobials and preservatives of natural origin. Thanks to their antimicrobial properties against pathogenic and food spoilage microorganisms, EOs have shown good potential for use as alternative food additives, also to counteract biofilm-forming bacterial strains, the spread of which is considered to be among the main causes of the increase in foodborne illness outbreaks. In this context, the aim of this study has been to define the antibacterial and antibiofilm profile of thyme (Thymus vulgaris L.) essential oil (TEO) against widespread foodborne pathogens, Salmonella enterica subsp. enterica serovar Typhimurium and Bacillus cereus. TEO chemical composition was analyzed through gas chromatography-mass spectrometry (GC-MS). Preliminary in vitro antibacterial tests allowed to qualitatively verify TEO efficacy against the tested foodborne pathogens. The subsequent determination of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values allowed to quantitatively define the bacteriostatic and bactericidal effects of TEO. To evaluate the ability of essential oils to inhibit biofilm formation, a microplate assay was performed for the bacterial biofilm biomass measurement. Results suggest that TEO, rich in bioactive compounds, is able to inhibit the growth of tested foodborne bacteria. In addition, the highlighted in vitro anti-biofilm properties of TEO suggest the use of this natural agent as a promising food preservative to counteract biofilm-related infections in the food industry.

Towards Green Strategies of Food Security: Antibacterial Synergy of Essential Oils from Thymus vulgaris and Syzygium aromaticum to Inhibit Escherichia coli and Staphylococcus aureus Pathogenic Food Isolates.

Foodborne diseases continue to represent an important public health issue. The control of food spoilage and pathogenic microorganisms is achieved mainly by synthetic chemicals, unfortunately associated to several undesirable aspects. The growing requirement for new and safe alternative strategies has resulted in the research of agents from natural sources with antimicrobial properties, such as essential oils (EOs). This study's purpose was to define the antibacterial profile of thyme (Thymus vulgaris) and cloves (Syzygium aromaticum) essential oils against both Gram-positive and Gram-negative important foodborne pathogenic bacteria. Gas chromatography mass spectrometry analysis was performed for EOs' chemical composition. Qualitative in vitro antimicrobial assays (i.e., agar well diffusion method and disk-volatilization method) allowed for verification of the efficacy of EOs, used individually and in binary combination and both in liquid and vapor phase, against Staphylococcus aureus and Escherichia coli food isolates. Minimal inhibitory concentrations and minimal bactericidal concentration values have been used to quantitatively measure the antibacterial activity of EOs, while the fractional inhibitory concentration index has been considered as a predictor of in vitro antibacterial synergistic effects. The microbiological tests suggest that thyme and cloves EOs, rich in bioactive compounds, are able to inhibit the growth of tested foodborne bacteria, especially in vapor phase, also with synergistic effects. Results provide evidence to consider the tested essential oils as promising sources for development of new, broad-spectrum, green food preservatives.

Chestnut Shell Tannins: Effects on Intestinal Inflammation and Dysbiosis in Zebrafish.

The aim of the present study was to test the possible ameliorative efficacy of phytochemicals such as tannins on intestinal inflammation and dysbiosis. The effect of a chestnut shell (Castanea sativa) extract (CSE) rich in polyphenols, mainly represented by tannins, on k-carrageenan-induced intestinal inflammation in adult zebrafish (Danio rerio) was tested in a feeding trial. Intestinal inflammation was induced by 0.1% k-carrageenan added to the diet for 10 days. CSE was administered for 10 days after k-carrageenan induced inflammation. The intestinal morphology and histopathology, cytokine expression, and microbiota were analyzed. The k-carrageenan treatment led to gut lumen expansion, reduction of intestinal folds, and increase of the goblet cells number, accompanied by the upregulation of pro-inflammatory factors (TNFα, COX2) and alteration in the number and ratio of taxonomic groups of bacteria. CSE counteracted the inflammatory status enhancing the growth of health helpful bacteria (Enterobacteriaceae and Pseudomonas), decreasing the pro-inflammatory factors, and activating the anti-inflammatory cytokine IL-10. In conclusion, CSE acted as a prebiotic on zebrafish gut microbiota, sustaining the use of tannins as food additives to ameliorate the intestinal inflammation. Our results may be relevant for both aquaculture and medical clinic fields.

In silico analysis of the antimicrobial activity of phytochemicals: towards a technological breakthrough.

BACKGROUND: The complications associated with infections from pathogens increasingly resistant to traditional drugs lead to a constant increase in the mortality rate among those affected. In such cases the fundamental purpose of the microbiology laboratory is to determine the sensitivity profile of pathogens to antimicrobial agents. This is an intense and complex work often not facilitated by the test's characteristics. Despite the evolution of the Antimicrobial Susceptibility Testing (AST) technologies, the technological breakthrough that could guide and facilitate the search for new antimicrobial agents is still missing. METHODS: In this work, we propose the experimental use of in silico instruments, particularly feedforward Multi-Layer Perceptron (MLP) Artificial Neural Network, and Genetic Programming (GP), to verify, but also to predict, the effectiveness of natural and experimental mixtures of polyphenols against several microbial strains. RESULTS: We value the results in predicting the antimicrobial sensitivity profile from the mixture data. Trained MLP shows very high correlations coefficients (0,93 and 0,97) and mean absolute errors (110,70 and 56,60) in determining the Minimum Inhibitory Concentration and Minimum Microbicidal Concentration, respectively, while GP not only evidences very high correlation coefficients (0,89 and 0,96) and low mean absolute errors (6,99 and 5,60) in the same tasks, but also gives an explicit representation of the acquired knowledge about the polyphenol mixtures. CONCLUSIONS: In silico tools can help to predict phytobiotics antimicrobial efficacy, providing an useful strategy to innovate and speed up the extant classic microbiological techniques.

Phytocompounds vs. Dental Plaque Bacteria: In vitro Effects of Myrtle and Pomegranate Polyphenolic Extracts Against Single-Species and Multispecies Oral Biofilms.

In the last decades, resistant microbial infection rate has dramatically increased, especially infections due to biofilm-producing strains that require increasingly complex treatments and are responsible for the increased mortality percentages compared with other infectious diseases. Considering that biofilms represent a key factor for a wide range of chronic infections with high drug tolerance, the treatment of biofilm-causing bacterial infections represents a great challenge for the future. Among new alternative strategies to conventional antimicrobial agents, the scientific interest has shifted to the study of biologically active compounds from plant-related extracts with known antimicrobial properties, in order to also evaluate their antibiofilm activity. In this regard, the aim of this study has been to assess the antibiofilm activity of polyphenolic extracts from myrtle leaf and pomegranate peel against oral pathogens of dental plaque, an excellent polymicrobial biofilm model. In particular, the in vitro antibiofilm properties of myrtle and pomegranate extracts, also in binary combination, were highlighted. In addition to inhibiting the biofilm formation, the tested polyphenolic extracts have been proven to destroy both preformed single-species and multispecies biofilms formed by Streptococcus mutans, Streptococcus oralis, Streptococcus mitis, and Rothia dentocariosa oral isolates, suggesting that the new natural sources are rich in promising compounds able to counteract biofilm-related infections.

In vitro Synergy of Polyphenolic Extracts From Honey, Myrtle and Pomegranate Against Oral Pathogens, S. mutans and R. dentocariosa.

The increasing incidence rate of oral diseases, the wide spread of antimicrobial resistance, and the adverse effects of conventional antibiotics mean alternative prevention and treatment options are needed to counteract oral pathogens. In this regard, our study aims to evaluate the antibacterial activity of polyphenolic extracts prepared from acacia honey, myrtle leaves, and pomegranate peel against cariogenic bacteria, such as Streptococcus mutans and Rothia dentocariosa. The chemical-physical parameters of acacia honey and the RP-HPLC polyphenolic profile of pomegranate peel extract have been previously described in our studies, while the characterization of myrtle extract, performed by HPLC analysis, is reported here. All the extracts were used singly and in binary combinations to highlight any synergistic effects. Moreover, the extracts were tested in association with amoxicillin to evaluate their ability to reduce the effective dose of this drug in vitro. The values of minimal inhibitory concentrations and minimal bactericidal concentrations have been used to quantitatively measure the antibacterial activity of the single extracts, while the fractional inhibitory concentration index has been considered as predictor of in vitro anticariogenic synergistic effects. Finally, a time-kill curve method allowed for the evaluation of the bactericidal efficacy of the combined extracts. The microbiological tests suggest that acacia honey, myrtle, and pomegranate extracts are able to inhibit the cariogenic bacteria, also with synergistic effects. This study provides useful and encouraging results for the use of natural extract combinations alone or in association with antibiotics (adjuvant therapy) as a valid alternative for the prevention and treatment of oral infectious diseases.

In Vitro Antibacterial Activity of Pomegranate Juice and Peel Extracts on Cariogenic Bacteria.

AIM: To evaluate the antimicrobial activity of hydroalcoholic extracts of pomegranate (Punica granatum L.) peel and juice, against the microorganisms considered the main etiologic agents of dental caries. METHODS: The values of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined against Streptococcus mutans Clarke ATCC® 25175™ strain and Rothia dentocariosa clinical isolate. RESULTS: Peel extracts inhibit effectively the growth and survival of S. mutans ATCC 25175 strain and R. dentocariosa clinical isolate with MIC and MBC values of 10 µg/µl and 15 µg/µl, respectively. Furthermore, the pomegranate juice extract showed high inhibitory activity against S. mutans ATCC 25175 strain with a MIC value of 25 µg/µl and a MBC value of 40 µg/µl, whereas, against R. dentocariosa, it has displayed a moderate inhibitory activity, with MIC and MBC values of 20 µg/µl and 140 µg/µl, respectively. CONCLUSIONS: In vitro microbiological tests demonstrate that the hydroalcoholic extracts of pomegranate juice and peel are able to contrast the main cariogenic bacteria involved in tooth decay. Although being preliminary data, our results suggest that pomegranate polyphenolic compounds could represent a good adjuvant for the prevention and treatment of dental caries.