Rhodomyrtus tomentosaleaf has been traditionally used to treat many infections. This plant species has been documented to possess a wide spectrum of pharmacological effects. This study aimed to determine the effects ofRhodomyrtus tomentosaleaf extract and its potent purified compound, rhodomyrtone, onStreptococcus pneumoniaevirulence factors including biofilms, capsule formation, and invasiveness which play important roles in infections. Ethanol leaf extract and rhodomyrtone demonstrated excellent antibacterial activity againstS. pneumoniaewith minimal inhibitory concentration (MIC) ranging from 16-32 mu g/ml and 0.125-1 mu g/ml, respectively. The ability of the extract and rhodomyrtone to prevent biofilm formation and eradicate mature biofilms was assessed. The extract and rhodomyrtone at 1/8 x MIC significantly inhibited biofilm formation in all clinical isolates (P < 0.05). The viability of 8-day biofilm-grown cells significantly decreased following the treatment with the extract and rhodomyrtone at 16 x MIC. 40-90% reduction in the bacterial adhesion and invasion to A549 human alveolar epithelial cells was observed after challenging with the extract and rhodomyrtone, compared with the control within 60 min. Increase in 90-99% phagocytosis of the bacterial cells by RAW264.7 macrophage cell line was detected following the treatment with the extract and rhodomyrtone at 1/2 x MIC, compared with the control. The results suggested potential medicinal benefits of the extract and rhodomyrtone for the treatment of pneumococcal infections.