Bacterial biofilms can harm humans in hospital environments, the food industry, and drinking water systems. The study aimed to examine the antibiofilm activity of Jatropha maheshwarii as an endemic plant. Biofilms were formed by Staphylococcus aureus and Pseudomonas aeruginosa on the surface of medical biomaterials. Moreover, time-dependent eradication of biofilms performed in polystyrene 96-well culture microplates was examined and expressed as minimal biofilm eradication concentration. The tested staphylococcus aureus formed the highest antibiofilm activity against biofilm at 77% in aqueous extract and the ethanol extract at 73% in different concentrations (µg/ml). The application of antibiofilm activity spans various fields, including medicine, the food industry, and environmental management.