This study aimed to evaluate the antibiofilm activity and toxicity of the glycolic extract of Persea americana "P.americana" over multidrug-resistant strains of Acinetobacter baumannii "A. baumannii" as alternative therapy to be investigated. A bacterial inoculum of each bacterial strain (4a, 5a, 9a, 12a, ATCC 19606) of A.baumannii was prepared and adjusted by the spectrophotometer. The microdilution broth method was performed to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). P.americana glycolic extracts were obtained of the tree stalk and leaves. The biofilm viability was tested by MTT assay after 5min exposure. The toxicity of the extracts was tested by invertebrate model Galleria mellonella. The data were analyzed by ANOVA, Tukey test and log-rank method (α=0.05). The extract showed an inhibitory and bactericidal action over all the tested strains with the lowest MIC value observed for the reference strain (3.12mg/mL). The extract did not demonstrate toxicity in any of the tested concentrations (12.5, 25 and 50mg/mL) in Galleria mellonella larvae, with a survival percentage above 80% after 168h. The glycolic extract of P.americana has microbicidal and antibiofilm activity on multidrug-resistant clinical strains of A.baumannii and showed low toxicity for the invertebrate model G.mellonella.