Central venous catheter-related bloodstream infection (CRBSI) is a huge public health concern with considerable impact on mortality and health costs. A three years observational study enrolling three tertiary hospitals located at Lisbon (Portugal) was designed to identify the major etiological agents of CRBSI, their biofilms on CVC and antimicrobial resistance (AMR) profiles. Etiological agents of CRBSI were isolated from the central venous catheter (CVC) and blood (HC) of 58 patients, being the most prevalent monomicrobial (Staphylococcus) and one case of dualmicrobial infection (E. cloacae complex/ C. parapsilosis) subjected to whole genome sequencing to confirm CRBSI and further characterize their resistome.