The antimicrobial activity of various extracts from Lactuca saligna L. (willow-leaf lettuce) was investigated in this work. Leaves and roots were collected from the Saatly district in Azerbaijan, and both aqueous and ethanolic extracts were prepared. The antimicrobial effects were evaluated using the disk-diffusion method against a selection of bacterial and fungal strains, including Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus anthracoides, and Candida albicans. Results showed that the aqueous root extract exhibited the most potent antibacterial activity, particularly against E. coli and K. pneumoniae, while the ethanolic root and leaf extracts demonstrated moderate activity against B. anthracoides and C. albicans. The study provides evidence supporting the traditional medicinal use of L. saligna for treating various infections. Furthermore, the findings suggest that L. saligna could be a promising source of natural antimicrobial agents, highlighting the need for further research to isolate and identify the active compounds responsible for its antimicrobial properties. This work contributes to the exploration of plantbased alternatives in the search for novel antimicrobial substances, particularly against drug-resistant pathogens.