The study aims to assess the frequency of ESKAPE pathogens and their antimicrobial resistance profile during the COVID-19 pandemic in a multidisciplinary hospital. Based on microbiological reports, the present study retrospectively evaluated the frequency and antimicrobial resistance (AMR) profile of ESKAPE microorganisms during 2020-2022 in a multidisciplinary hospital in Romania. There were reported 1117 bacterial isolates during the study and ESKAPE pathogens account for 33.5% of the strains. The specific AMR of ESKAPE pathogens cumulates 30.9%, while antimicrobial multidrug resistance characterizes 21% of all ESKAPE strains. The AMR profile of K. Pneumoniae, followed by S. aureus, and P. aeruginosa are alarming on ESKAPE surveillance. The most frequent bacterial isolates and antibiotic resistance reports were identified in the urology department. Based on the local AMR data we suggest the first-line antibiotic recommendations when ESKAPE pathogens are suspected. The directions of the near future antibiotic stewardship program focus on revising the clinical protocols of diagnosis and treatment in each department, increasing the bacteriological samples collection, improving the microbiological diagnostic techniques for AMR identification, and intensifying the epidemiological actions of surveillance, control, and education.