Daun gatal (Laportea aestuans) is an indigenous plant of Papua hich has been widely used for pain relief as traditional medication in the local community. The leaves were just picked then treated to cure for painful body. This treatment are giving itchy sense as an indication that medication is working on the body as local people assumed. The aim of the study was to determine the activity of antibacteria of daun gatal against Escherichia coli, Staphylococcus aureus and Salmonella typhi. Samples of daun gatal were collected from Biak Numfor Papua. The methods are started with filtering of daun gatal using 100 mash and extracted with ethanol. Extract was made in 250, 500, 750, 1000 ppm, with chloramphenicol as a control positive, while the test of antibacterial activity was done by disc diffusion method. The results showed that daun gatal has antibacterial activity against E. coli, S. aureus and S. typhi. Daun gatal extract (L. aestuans) is effective in inhibiting the growth of bacterias with zone inhibition of 1000 ppm extracts which were 8.55 mm (E. coli), 9.02 mm (S. typhi), dan 9.37 mm (S. aureus). Key words: Antibacteria, E. coli, L. aestuans, S. aureus and S. typhi.