| Summary: | The spread of bacterial resistance to antibiotics affects various areas of life. The aim of this study was to assess the occurrence of <i>Pseudomonas aeruginosa</i>, and other bacteria mainly from orders <i>Enterobacterales </i>and <i>Staphylococcus</i> in the pharmaceutical production sites, and to characterize isolated strains in the aspects of antibiotic resistance, biofilm formation, and presence of genes encoding virulence factors. Genes encoding selected virulence factors were detected using PCR techniques. Antimicrobial susceptibility testing was applied in accordance with the EUCAST recommendations. A total of 46 <i>P. aeruginosa </i>strains were isolated and 85% strains showed a strong biofilm-forming ability. The qualitative identification of genes taking part in <i>Quorum Sensing</i> system demonstrated that over 89% of strains contained<i> lasR</i> and <i>rhlI</i> genes. An antimicrobial susceptibility testing revealed nine strains resistant to at least one antibiotic, and two isolates were the metallo-β-lactamase producers. Moreover, the majority of <i>P. aeruginosa</i> strains contained genes encoding various virulence factors. Presence of even low level of pathogenic microorganisms or higher level of opportunistic pathogens and their toxic metabolites might result in the production inefficiency. Therefore, the prevention of microbial contamination, effectiveness of sanitary and hygienic applied protocols, and constant microbiological monitoring of the environment are of great importance.
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