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OBJECTIVES The Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) is a global surveillance programme monitoring the in vitro activity of a panel of antimicrobial agents against clinically important bacterial isolates. Data for Gram-positive and Gram-negative isolates collected in Eastern Europe between 2011 and 2016 are presented here. METHODS Minimum inhibitory concentrations (MICs) were determined by the broth microdilution method using CLSI guidelines. Antimicrobial susceptibility was assessed using EUCAST breakpoints. RESULTS Nine Eastern European countries submitted 4289 isolates. Among Acinetobacter baumannii, resistance to levofloxacin, amikacin and meropenem was 77.5%, 63.4% and 62.2%, respectively. Multidrug resistance among A. baumannii was higher in 2015 than in previous years (44.1% in 2011 and 71.0% in 2015), decreasing to 51.7% in 2016. The multidrug resistance percentage for Pseudomonas aeruginosa was 26.9% and was relatively stable over time. The percentage of extended-spectrum β-lactamase (ESBL)-positive isolates among Escherichia coli and Klebsiella pneumoniae was 20.1% and 55.7%, respectively. Resistance to amikacin, meropenem and tigecycline was low among E. coli and K. pneumoniae and the ESBL-producing subset (≤5.9%). Among Staphylococcus aureus isolates, 36.7% were methicillin-resistant (MRSA); percentages varied year-on-year. No S. aureus isolates, including MRSA, were resistant to linezolid, vancomycin or tigecycline. Among Enterococcus faecium isolates, resistance was 22.6% to vancomycin and 2.3% to linezolid; no isolates were resistant to tigecycline. CONCLUSION This study shows low resistance to meropenem and tigecycline among Enterobacteriaceae isolates and continued activity of linezolid, vancomycin and tigecycline against Gram-positive organisms. However, antimicrobial resistance continues to be problematic in Eastern Europe and requires continued surveillance.