Antimicrobial, Anti-Biofilm, Anti-Quorum Sensing and Cytotoxic Activities of Thymbra spicata L. subsp. spicata Essential Oils.

Background/Objectives: Essential oils of Thymbra spicata subsp. spicata are known for their rich phytochemical content and bioactive properties. This study aimed to evaluate the antimicrobial, anti-biofilm and anti-quorum sensing, as well as the cytotoxic activities of T. spicata subsp. spicata essential oils (TS-EO1 and TS-EO2) obtained from two different localities in Türkiye, along with a detailed chemical composition analysis. Methods: TS-EO1 and TS-EO2 were obtained by the hydrodistillation method and analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) to determine their phytochemical profiles. Antimicrobial activities were assessed against Gram-positive and Gram-negative bacteria, and fungal strains were assessed using the broth microdilution method. Anti-biofilm and anti-quorum sensing activities were evaluated using Pseudomonas aeruginosa PAO1 and Chromobacterium violaceum ATCC 12472, respectively. Cytotoxic properties were tested on four cell lines (A549, MCF-7, U87MG, and L929) using the MTT assay. Results: Both essential oil samples were rich in carvacrol (54.3% and 54.1%), followed by p-cymene and γ-terpinene. The essential oils exhibited significant antimicrobial activity, particularly against Staphylococcus aureus (6.25 mg/mL) and Candida parapsilosis (0.20 mg/mL). Sub-MIC concentrations significantly inhibited biofilm formation and quorum sensing. Both samples showed moderate cytotoxic properties against human cancer cell lines, particularly A549 (IC50: 116.3 and 134.4 μg/mL, respectively). Conclusions: This study showed that T. spicata subsp. spicata essential oils have significant antimicrobial, anti-biofilm, and anti-quorum sensing properties against various bacteria and fungi, along with moderate cytotoxic effects, indicating their medicinal and pharmaceutical potentials. This is the first study which revealed anti-biofilm and anti-quorum sensing properties of T. spicata essential oils to our knowledge.