Natural extracts containing high polyphenolic concentrations may act as good antimicrobials for their antibacterial and antibiofilm activity. The present research characterizes two hydro-organic extracts with high polyphenolic content, obtained from the shrub Cytisus scoparius as antipathogenic candidates. As a result of their own composition, both extracts, LE050 and PG050, have shown pronounced bioactivities with potential uses, especially in agricultural, livestock production, food manufacturing, and pharmaceutical industries. Polyphenolic compounds were extracted by using adjusted hydro-organic solvent mixtures. These extracts’ in vitro antimicrobial activity was evaluated on Gram-positive and Gram-negative pathogenic bacteria, giving special attention to those involved in food contamination. Due to this, the biofilm dispersion was assessed on Listeria monocytogenes, Staphylococcus aureus and Pseudomonas aeruginosa. The extracts showed antimicrobial activity against the pathogenic species tested, presenting IC50 values between 0.625–20% v/v. Different behaviors have been detected between both extracts, probably linked to their distinct polyphenol composition, being LE050 extract the one with most promising bioactive applications. Finally, the results from the biofilm dispersion assays reveal that the extracts exhibit a good antibiofilm activity against the pathogenic bacteria tested.

Infections caused by multidrug-resistant bacteria are becoming increasingly frequent and sometimes difficult to treat due to the limited number of antibiotics active against them. In addition, they can spread between countries and/or continents, which is a problem of great relevance worldwide. It is, therefore, urgent to find alternatives to treat infections caused by multidrug-resistant bacteria. This study aimed at exploring a possible therapeutic alternative in the fight against antibiotic resistance. Based on the known antibacterial capacity of polyphenols, we tested the antimicrobial activity of a polyphenolic extract of Albariño white grape marc on clinical strains since research on such bacteria has been very scarce until now. First, the extract was obtained using a medium-scale ambient temperature (MSAT) system, which is an efficient and sustainable extractive method. The determinations of the polyphenolic content of the extract and its antioxidant capacity showed good results. Using chromatographic and mass spectrometric tools, 13 remarkable polyphenols were detected in the extract. The antibacterial activity of our grape marc extract against nineteen clinical strain isolates, some of which are multidrug-resistant, was evaluated by means of the calculation of half of the maximum inhibitory concentration (IC50) and the value of the minimum bactericidal concentrations (MBCs). In conclusion, the extract showed effectiveness against all clinical strains tested, regardless of their level of antibiotic resistance, and shows promise in the fight against antibiotic resistance.

Background/Objectives: The increasing prevalence of antimicrobial-resistant bacteria highlights the need for improved methodologies to evaluate antimicrobial activity beyond conventional minimum inhibitory concentration testing. While resazurin-based assays are widely used for minimum inhibitory concentration determination due to their simplicity and sensitivity, minimum bactericidal concentration assessment still relies on labor-intensive colony-forming unit counting. The objective of this study was to develop and validate a resazurin-based microwell assay capable of determining both the minimum inhibitory concentration and the minimum bactericidal concentration without routine plate counting, thereby simplifying bactericidal evaluation. Methods: A two-step resazurin-based fluorescence assay was designed and performed in microplates. After determining the minimum inhibitory concentration using resazurin as a metabolic indicator, well-showing inhibited bacterial growths were subjected to a regrowth phase by transferring aliquots into fresh antimicrobial-free medium containing resazurin. This additional step allowed discrimination between reversible metabolic inhibition and irreversible bacterial death. The method was evaluated using ciprofloxacin and chloramphenicol against four bacterial species: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Minimum bactericidal concentration values obtained using this assay were compared with those obtained through conventional colony counting on agar plates. Results: Minimum bactericidal concentration values obtained using the two-step fluorescence assay were fully concordant with the conventional colony-forming unit counting method for all tested antibiotics and bacterial species. Conclusions: The proposed two-step resazurin-based microwell assay represents a rapid, reliable, and less labor-intensive alternative for the determination of both the minimum inhibitory concentration and the minimum bactericidal concentration, with potential applications in clinical and industrial microbiology laboratories.

The production of β-lactamases is the main mechanism underlying carbapenem resistance. This study combined in silico and in vitro approaches to identify potential polyphenols as carbapenemase inhibitors. Molecular docking, molecular dynamics, and ADMET prediction were performed to assess the binding affinity, stability, and safety of quercetin, kaempferol, caffeic acid, and 3,4-dihydroxybenzoic acid against KPC-2, NDM-1, and OXA-48 carbapenemases. In vitro antibacterial assays and checkerboard analyses were conducted against Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa to assess antibacterial and synergistic effects. Then, the inhibition of the β-lactam hydrolytic activity was confirmed. In silico results showed that quercetin, kaempferol, and caffeic acid exhibited strong binding affinity and consistent stability towards the targets. Therefore, quercetin and kaempferol showed the strongest affinities (−8.0 kcal/mol) and stable interactions with key catalytic residues. ADMET profiles indicated good pharmacokinetic behavior and low acute toxicity. In vitro assays revealed that the polyphenols exhibited MIC values ranging from 12.5 to 25 mg/L and MBC values of 25–50 mg/L. Combined with cefotaxime, they enhanced bacterial susceptibility and inhibited β-lactam hydrolysis, with quercetin achieving complete inhibition at 200 mg/L. These findings highlight the potential of the four polyphenols as natural β-lactamase inhibitors. Further enzyme kinetics and in vivo studies are needed to confirm their therapeutic relevance.

This study evaluates the potential role of two extracts derived from green and roasted coffee in managing Type 2 Diabetes (T2D). The phytochemical analysis revealed that the roasted coffee extract (RCE) contains higher levels of flavonoids and tannins, while the green coffee extract (GCE) seems to be richer in phenolic acids. No differences in the antioxidant activity of both extracts were observed. The study demonstrated that GCE exhibits stronger prebiotic potential by significantly enhancing the growth of beneficial probiotic bacteria when compared to the untreated sample. Both extracts inhibited α-amylase and α-glucosidase enzymes, with RCE demonstrating a better performance than other commercial treatments for T2D. Glucose uptake assays on yeast cells demonstrated that both extracts enhance glucose transport, particularly at low glucose concentrations, reducing supernatant glucose levels by 60–80%. Notably, GCE maintained its effectiveness even at the highest glucose concentrations tested. These findings suggest that coffee extracts, particularly GCE, may be useful as nutraceuticals for potentially regulating glucose metabolism and gut microbiota in T2D management.