Background/Objectives: This study investigates the phytochemical profile, essential oil composition, and bioactivities—including antioxidant, antimicrobial, antibio-film, and anti-quorum sensing (QS) activities—of four Heracleum L. species (H. crenatifolium Boiss, H. paphlagonicum Czeczott, H. sphondylium subsp. montanum Schleich. ex Gaudin, and H. pastinacifolium subsp. incanum (Boiss. & A.Huet) P.H.Davis). Methods: Total phenolic and flavonoid contents were quantified using the Folin–Ciocalteu and aluminum chloride colorimetric methods, respectively. Essential oils were extracted by hydrodistillation and analyzed via Gas Chromatography–Flame Ionization Detector (GC–FID) and Gas Chromatography–Mass Spectrometry (GC–MS), while Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) evaluated chemical variability among the species. Antioxidant activities were assessed using DPPH and ABTS free radical scavenging assays. Antimicrobial activity was assessed using the broth microdilution method to determine Minimum Inhibitory Concentration (MIC) values, while antibiofilm activity was evaluated using an in vitro microplate-based biofilm model against Pseudomonas aeruginosa PAO1. Anti-QS activity was analyzed using a disc diffusion assay with Chromobacterium violaceum ATCC 12472 as the reporter strain. Results: It was observed that the amounts of total phenolic compounds and total flavonoids were higher in root extracts than in aerial parts extracts for the four species in this study (H. sphondylium subsp. montanum excluding phenolic content). In the analysis of essential oil, it was determined that the major component in the roots was mostly myristicin, and in the fruits it was mostly octyl acetate. Phenolic and flavonoid contents were positively correlated with antioxidant activity. Methanol and n-hexane extracts of H. pastinacifolium (aerial parts) and n-hexane extracts of H. paphlagonicum (root) exhibited notable antimicrobial activity, primarily against Gram-positive bacteria, but none of the extracts showed activity against Klebsiella pneumoniae ATCC 13383 or P. aeruginosa ATCC 27853. Among methanol extracts, H. pastinacifolium (aerial parts) exhibited the highest antibiofilm activity (73.2%), while H. paphlagonicum (aerial parts) showed the highest activity among n-hexane extracts (75.5%). All n-hexane extracts exhibited anti-QS activity, whereas the methanol extracts showed no activity. Conclusions: These findings underscore the chemical diversity and bioactive potential of Heracleum species, contributing to the chemotaxonomic understanding of the genus and supporting their potential applications in medicine and industry. To our knowledge, this is the first study that reveals the antibiofilm and anti-QS properties of these Heracleum species.

Background/Objectives: In this study, nanoemulsions containing Thymus vulgaris (TV) and Rosmarinus officinalis (RO) essential oils (EOs) were prepared and characterized, and their antibacterial, antifungal, and antibiofilm activities were evaluated against some oral pathogens. Methods: Three nanoemulsions containing 5% TV-EO (F1), 5% RO-EO (F2), and a combination of 2.5% TV-EO and 2.5% RO-EO (F3) were prepared by emulsification followed by high-speed homogenization. Results: Gas chromatography–mass spectrometry coupled with flame ionization detection (GC-MS-FID) analysis revealed linalool (79.51%) as the major constituent of TV-EO and eucalyptol (58.13%), together with camphor (10.80%), as the predominant compounds of RO-EO. All nanoemulsions exhibited nanosized droplets, low polydispersity index (PDI) values (<0.3), and moderate to high colloidal stability. Transmission electron microscopy (TEM) analysis revealed predominantly spherical droplets in all formulations (F1–F3). All formulations exhibited antimicrobial activity against Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, and Candida albicans. Among the tested formulations, F2 showed the strongest antibacterial activity, whereas F3 exhibited the highest antibiofilm activity against mature S. mutans biofilms, achieving approximately 80% inhibition, comparable to chlorhexidine (CHX). Although F3 possessed the smallest droplet size, its antimicrobial activity was lower than that of F2, suggesting that biological activity was influenced not only by droplet size but also by EO composition. Conclusions: These findings demonstrate the potential of TV-EO and RO-EO nanoemulsions as promising natural alternatives for mouthwash formulations and highlight the importance of EO composition in determining antimicrobial and antibiofilm efficacy.