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Background: Arbekacin is a broad-spectrum aminoglycoside with activity against some Gram-positive and Gram-negative bacteria. Methods: Arbekacin minimum inhibitory concentration (MIC) values were determined for 296 drug-resistant Gram-negative bacilli, and compared to previously determined plazomicin, amikacin, gentamicin, and tobramycin MIC values. Results: The MIC values required to inhibit 50% and 90% of isolates (MIC50 and MIC90, respectively) were 16 and >128 μg/ml, respectively. Conclusions: Arbekacin showed similar MIC50 values to amikacin and gentamicin, a lower MIC50 value than tobramycin, and a higher MIC50 value than plazomicin.