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Introduction:The present work aims to study the chemical composition of Pteridium aquilinum (L.) Kuhn essential oil and its antibacterial activity against three important phytopathogenic Gram-negative bacteria: Erwinia amylovora, Pectobacterium carotovorum subsp. carotovorum, and Pseudomonas savastanoi pv. savastanoi. Methods: The chemical composition of P. aquilinum L. essential oil produced by hydrodistillation was determined by gas chromatography-mass spectrometry. The antibacterial activity was tested using disk diffusion method and by determination of minimum inhibitory concentration values. The major components were linalool (10.29%), carvacrol (8.15%), benzaldehyde (5.95%), 2-undecanone (5.32%), and cuminaldehyde (4.57%). Results: The essential oil tested revealed a powerful antibacterial effect against all tested strains, with inhibition zone diameters ranging from 32.0 ± 0.58 to 33.7 ± 0.88 mm. Discussion: P. aquilinum EO contained 32.86% of oxygenated monoterpenes, which are known for their very powerful antimicrobial activities. The minimum inhibitory concentration values showed that P. aquilinum essential oil has very strong activity against E. amylovora (0.625 μl/ml), followed by P. carotovorum subsp. carotovorum (2.50 μl/ml) and P. savastanoi pv. savastanoi (5.00 μl/ml). The results obtained could contribute to the development of new potential agents for the control of bacterial diseases.