This study provides a detailed description of the distribution of non-fermentative gram-negative bacteria (NFGNB) collected in water sources (tap water and water used for haemodialysis and bronchoscope flushing) from different wards of a tertiary care hospital. The aim is to identify risk practices for patients or to alert clinicians to the possible contamination of environment and medical devices. The resistance profile of NFGNB environmental isolates has shown that more than half (55.56%) of the strains isolated were resistant to one or more antibiotics tested in different antimicrobial categories. In particular, 38.89% of these strains were multidrug resistant (MDR) and 16.67% were extensively drug resistant (XDR). The most prevalent bacterial species recovered in water samples were Pseudomonas aeruginosa, Pseudomonas fluorescens, Ralstonia pickettii and Stenotrophomonas maltophilia. Analysis of antibiotic resistance rates has shown remarkable differences between Pseudomonadaceae (P. aeruginosa and P. fluorescens) and emerging pathogens, such as S. maltophilia and R. pickettii. Multidrug resistance can be relatively common among nosocomial isolates of P. aeruginosa, which represent the large majority of clinical isolates; moreover, our findings highlight that the emergent antibiotic resistant opportunistic pathogens, such as R. pickettii and S. maltophilia, isolated from hospital environments could be potentially more dangerous than other more known waterborne pathogens, if not subjected to surveillance to direct the decontamination procedures.