OBJECTIVES: The period of the COVID-19 pandemic had a significant impact on the healthcare system, including its effect on compliance with the established procedures of a rational antibiotic policy, especially in the context of nosocomial pneumonia, where it was very difficult to distinguish a possible bacterial superinfection from a severe inflammatory reaction caused by the SARS-CoV-2 virus. The aim of the present study was to analyze the antimicrobial resistance of bacterial pathogens isolated from the lower respiratory tract and their clonality in intensive care patients in 2022 and to compare it with the previous COVID-19 period. MATERIAL AND METHODS: Bacterial strains isolated from the lower respiratory tract (LRT) of patients hospitalized at the Department of Anaesthesiology, Resuscitation and Intensive Care, Olomouc University Hospital (DARIC) over a three-year period (January 1, 2020 - December 31, 2022) were included in the study. The susceptibility to antibiotics was determined by the standard microdilution method according to the EUCAST criteria, and selected isolates were compared using pulsed-field gel electrophoresis (PFGE). RESULTS: The resistance of the most common bacterial pathogens isolated from the LRT of patients hospitalized at DARIC did not change significantly during the COVID-19 (2020-2021) and post-COVID-19 (2022) periods, with the exception of Serratia marcescens and Enterococcus faecium species. These two showed an increase in the number of strains during the COVID-19 pandemic, as well as a significant increase in the proportion of resistant strains. In the case of Serratia marcescens, there was a subsequent decrease in the number of isolates and their resistance in 2022. For Enterococcus faecium, the total number of isolates also decreased significantly, but the frequency of vancomycin-resistant isolates (VRE) continued to increase. During the COVID-19 pandemic, increased VRE detection can be linked to proven clonal spread, but significant clonality was no longer confirmed in 2022. Comparison of similarity by PFGE in other bacterial species also did not reveal significant horizontal transmission between patients in the post-COVID-19 period, as most isolates (85%) showed a unique restriction profile. CONCLUSIONS: The results indicate that the frequency and antimicrobial resistance of the majority of the most common bacterial pathogens from the LRT of patients hospitalized at DARIC in the post-pandemic period remain comparable to the time before and during the COVID-19 pandemic outbreak. An exception is Enterococcus faecium, which showed an increase in vancomycin resistance in both the COVID-19 and the post-COVID-19 periods.