Biofilm formation on indwelling devices promotes antimicrobial resistance and complicates management of ICU infections; comprehensive profiling of biofilm-producing pathogens in ICU settings is needed to inform infection control and stewardship. Cross-sectional study (June–September 2023) of clinical isolates from ICU patients with sepsis, pneumonia, urinary tract infection, or wound infection at Sultan Agung Islamic Hospital, Semarang. Sixty-two isolates from sputum (endotracheal aspirates), pus/wound swabs, urine, and blood were identified by standard culture and biochemical methods. Biofilm production was assessed by the 96‑well microtiter plate crystal violet assay. All isolates produced biofilm: 50/62 (80.6%) strong, 9/62 (14.5%) moderate, and 3/62 (4.8%) weak. Gram‑negative organisms predominated: Klebsiella pneumoniae 18/62 (29.0%), Pseudomonas aeruginosa 17/62 (27.4%), and Escherichia coli 10/62 (16.1%). All P. aeruginosa isolates (17/17; 100%) exhibited strong biofilm formation across clinical conditions. K. pneumoniae was the most frequent cause of pneumonia (15/62; 24.2%), with 13/15 (86.7%) strong biofilm producers. ICU isolates showed a high prevalence of strong biofilm production, primarily among Gram‑negative pathogens. These findings support the implementation of targeted surveillance, strict device care protocols, and antimicrobial stewardship strategies to address biofilm-associated resistance; larger multicenter longitudinal studies are recommended.
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