Exploration of new source of antibiotics to combat multidrug-resistant bacteria is urgently needed. Indonesia as archipelago country has a wide variety of marine organisms with potential as source of new antibacterial compounds against MDRO. Aims of the study were to isolate sponge-associated fungi from sponge Cinachyrella sp. collected from Pandang Island, North Sumatera, Indonesia, to screen potential fungi against clinical pathogenic MDR bacteria, to identify the potential fungi; and to determine the best cultivation time for antibacterial production. Nine spongeassociated fungi were successfully isolated. Result of agar plug method showed fungus PDSP 5.7 was the most potential candidate which inhibited ESBL Escherichia coli, Salmonella enterica ser. Typhi, MRSA, and Staphylococcus haemolyticus strain MDR. This fungus had 100% similarity to Trichoderma reesei. In malt extract broth, T. reesei PDSP 5.7 had stationary phase from day 12 to day 18. In addition, the widest antibacterial was performed by extract from day 15. Furthermore, fungal extract showed best antibacterial activity against S. enterica ser. Typhi strain MDR with inhibition value of 14.72 ± 0.07 mm 2 .