This study aimed to investigate the antibacterial activity of essential oils from Cupressus sempervirens (CSEO) and Eucalyptus globulus (EGEO) against clinical isolates of colistin-resistant E. coli and methicillin-resistant M. sciuri recovered from diseased food-producing animals in Tunisia. The chemical compositions of both essential oils were analyzed using Gas chromatography-mass spectrometry (GC-MS). Antibacterial activity was determined through disk diffusion and microdilution assays, while the fractional inhibitory concentration index (FICI) was used to measure the synergistic effect of CSEO and colistin. The major constituents of CSEO were α-pinene (59.03%), δ-3 Carene (21.48%), D-limonene (5.62%) and α-thujene (3.08%), while EGEO contained eucalyptol (52.1%), α-pinene (26.51%), γ-terpinene (4.81%) and D-limonene (3.54%). The inhibition zone diameters (IZDs) against E. coli and M. sciuri ranged from 6 to 29 mm for CSEO and 9.3 to 28.6 mm for EGEO. Minimum inhibitory concentrations (MICs) showed a range of 4 to 512 µg/mL for CSEO against M. sciuri and 2 to 426.66 µg/mL for EGEO. For E. coli, MICs ranged from 170.66 to 512 µg/mL for CSEO and 53.33 to 512 µg/mL for EGEO. The checkerboard test demonstrated a synergistic effect between CSEO and colistin against two colistin-resistant E. coli isolates, with FICI 3- to 32-fold lower than the MICs of the individual compounds. This study highlights the potency of essential oils from CSEO and EGEO against antibiotic-resistant M. sciuri and E. coli strains from animals in Tunisia and provides evidence of the synergistic effect between CSEO and colistin against clinically relevant colistin-resistant E. coli isolates.
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