Fourteen seaweeds collected from the intertidal zone of Southwest coast of India were tested against ten human pathogen bacteria and one human pathogen fungus using the well diffusion test in the casitone agar medium. The species used in the present study include five Chlorophyta (Bryopsis plumosa, Ulva fasciata, Acrosiphonia orientalis, Chaetomorpha antennina, Grateloupia filicina), five Rhodophyta (Hypnea pannosa, Gracilaria corticata, Centroceras clavulatum, Portieria hornemannii, Cheilosporum spectabile) and four Phaeophyta (Padina tetrastromatica, Sargassum wightii, Stocheospermum marginatum, Chnoospora bicanaliculata). Of these, seven species were determined to be highly bioactive and screened on the multiresistant pathogens. We found that drying process has eliminated the active principles in the seaweeds. In the present study, methanol:toluene (3∶1) was found to be the best solvent for extracting the antimicrobial principles from fresh algae. However, the ethanolic extract showed no antibacterial activity.Acrosiphonia orientalis showed activity against 70% of the tested organisms.Stocheospermum marginatum was the only seaweed that showed activity againstKlebsiella pneumoniae. The extract fromGracilaria corticata was highly active againstProteus mirabilis, a Gram negative pathogenic bacterium. The present findings revealed that the tested seaweeds were highly active against Gram negative bacteria than Gram positive bacteria. The antimicrobial principle from seaweed was found to be a lipophilic compound. The compound was stable over a wide range of temperature (30–60 °C). The active principles of highly active seaweedsAcrosiphonia orientalis andStocheospermum marginatum were bactericidal.