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This study aimed to characterize MDR <jats:italic>A. baumannii<\/jats:italic> isolates for their biofilm\u2010forming capabilities and the presence of common biofilm\u2010related genes at a tertiary care university hospital in Nepal. In addition, it assessed the efficacy of various compounds, particularly essential oils, in inhibiting biofilm formation. Identification and antibiotic sensitivity testing of <jats:italic>A. baumannii<\/jats:italic> isolates from clinical specimens were conducted according to the guidelines of the American Society for Microbiology. Isolates were screened for motility profiles, biofilm production in a microtiter plate assay, and the presence of biofilm\u2010related gene(s) by conventional polymerase chain reaction. The ability of cinnamaldehyde, ethylenediaminetetraacetic acid (EDTA), Tween 80, amino acids (glycine and glutamic acid), and natural plant extracts to inhibit biofilm formation was also tested using the microtiter plate system. Out of the total 200 <jats:italic>A. baumannii<\/jats:italic> isolates, 195 were MDR, with 192 able to produce biofilms. Among them, 83.1% were strong biofilm producers. In this study, 42.0% and 66.2% of the isolates exhibited twitching motility and surface\u2010associated motility, respectively. Thirty MDR <jats:italic>A. baumannii<\/jats:italic> isolates from medical devices contained biofilm\u2010related genes <jats:italic>csuE, ompA, bap,<\/jats:italic> and <jats:italic>b<\/jats:italic><jats:italic>l<\/jats:italic><jats:italic>a<\/jats:italic><jats:sub>PER\u22121<\/jats:sub>, in 90.0%, 53.3%, 46.6%, and 26.6% of strains, respectively. Cinnamaldehyde (0.875\u2009mg\/mL) was the most effective compound, inhibiting biofilm formation by 77.3%, followed by ethanolic extract of onion (77.2%), 0.5% Tween 80 (76.8%), and essential oil of ginger (70.8%). The majority of <jats:italic>A. baumannii<\/jats:italic> clinical isolates were strong biofilm producers and often possessed the biofilm\u2010related genes <jats:italic>csuE<\/jats:italic> and <jats:italic>ompA<\/jats:italic>. Essential oils at 200\u2009mg\/L, along with Tween 80, were the most effective (\u2265 67%) at inhibiting the formation of biofilms. These findings help to understand biofilm production and provide valuable insights into MDR <jats:italic>A. baumannii<\/jats:italic> isolates in this clinical setting.<\/jats:p>","DOI":"10.1155\/ijm\/5749982","type":"journal-article","created":{"date-parts":[[2024,12,28]],"date-time":"2024-12-28T08:50:01Z","timestamp":1735375801000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Characterization and Biofilm Inhibition of Multidrug\u2010Resistant <i>Acinetobacter baumannii<\/i> 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