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Objective. This study aims to investigate the bacterial biofilm-inhibitory effect of mushroom extracts. Methods. Mushrooms were collected from Arabuko-Sokoke and Kakamega forests and identified using morphological and molecular approaches. Auricularia auricula-judae, Microporus xanthopus, Termitomyces umkowaani, Trametes elegans, and Trametes versicolor were extracted by chloroform, 70% ethanol, and hot water. Extracts were tested against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus (ATCC25923). Data were analyzed using SPSS ver. 20.0. Results. Chloroform, 70% ethanol, and hot water extracts of A. auricula-judae (50 μg/mL) showed statistically significant antibiofilm activities against P. aeruginosa, E. coli, and S. aureus (p ≤ 0.05). M. xanthopus extracts (250 μg/mL) revealed significantly significant antibiofilm activities against each test bacterium (p ≤ 0.05). All extracts of T. umkowaani (250 μg/mL) exhibited statistically significant antibiofilm activities against S. aureus only (p ≤ 0.05). Chloroform extract of T. elegans (250 μg/mL) showed the best antibiofilm activity (69.75 ± 0.01%) against S. aureus. All T. versicolor extracts (250 μg/mL) indicated the best antibiofilm activities against S. aureus. Conclusions. Being the first study of its kind to be conducted in Kenya, it added a novel concept to the body of knowledge already known about medical biotechnology research. It offers a fresh understanding of the various varieties of mushrooms found in Kenya, their potential biological function in the production of drugs, particularly those that combat drug resistance, and perhaps even a peek at their bioactive elements. Wild mushrooms, a hidden gem, might help to reopen the pipeline of new antibiotics that have been on the decline. However, further research is required to determine the potential mechanism(s) of action of the extracts that are in charge of the apparent antibiofilm activity. [ABSTRACT FROM AUTHOR]