Ascidians are marine biofouling animals with a proven ability to synthesis bioactive substances. This study investigated the bioactive potential of ascidian Lissoclinum fragile, found in the coastal waters of Tuticorin Southeast coast of India. Freshly collected ascidian soaked in methanol for five days and filtered through filter paper. The solvent was concentrated by rotary evaporator which reduced the pressure to give a dark brown gummy mass. The collected gummy mass was separately extracted

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... successively with acetone, n-butanol, chloroform, ethyl acetate and dichloromethane. These extracts were used for antimicrobial, heamolytic and cytotoxic assays. In antibacterial assay bacterial pathogen, S. typhi exhibit high zone of inhibition (14.1 ± 0.01 mm) against dichloromethane extract. In antifungal assay, fungal pathogen Penicillium species showed high zone of inhibition (12.3 ± 0.07 mm) against nbutanol extract. In haemolytic assay, n-butanol extract showed high heamolytic activity in chicken erythrocytes (64 HU), goat erythrocytes (16 HU) and cow erythrocytes (16 HU). In cytotoxic activity, nbutanol extract exhibited high LC 50 value (97 µg/ml) against brine shrimps. These results indicate that the ascidian L. fragile has remarkable antimicrobial, haemolytic, and cytotoxic activities.