Biofilms are the structure microbial that attach to different surfaces and enclave microorganisms in a matrix composed, protecting them from harsh conditions, including immune effectors and antibiotics. Therefore, this study aimed to assess the anti-biofilm activity of lavender, cloves, eucalyptus, rosemary, thyme, and oregano commercial essential oils, known for their biocidal properties against Gram-positive and Gram-negative ESKAPE isolates, exhibiting multiple drug resistance. The most efficient proved to be the oregano, thyme, and clove essential oils, which exhibited the lowest minimal biofilm eradication concentrations values against all bacterial species. Although lavender, eucalyptus, and rosemary had a weaker effect than the first three oils, they have also inhibited biofilm development at concentrations low enough to be considered effective. This experimental approach may open new perspectives for developing efficient strategies to combat the emergent threat of antibiotic resistance and to control the formation of microbial biofilms, being also a valuable source of bioactive compounds and new anti-biofilm drugs.