The distribution and antibacterial activities of bacteria isolated from fifty-five marine sponge samples collected from the Aegean Sea and the Sea of Marmara were investigated in the period between 2011 and 2013. The antibacterial activities of the methanolic extracts of marine sponge-associated bacteria tested against six pathogenic bacteria: Staphylococcus aureus SA1 and SA2, Brucella melitensis B37, Vibrio vulnificus GK23, Escherichia coli O157:H7 (ATCC 35150), and Salmonella enterica subsp. enterica serovar typhi (ATCC 167), using disk diffusion tests and minimum inhibitory concentration technique. The bacteria isolated from sponges and ambient seawater samples were identified using VITEK 2 Compact 30 automated microbial identification system. All bacterial extracts were exhibited antibacterial activity with various MIC values ranging from 7.8 mg/ mL to 1000 mg/mL against all pathogenic bacteria tested. The antibacterial efficacy rates found to be higher in the Aegean Sea than the Sea of Marmara samples. Fifty-five sponge samples belonging to fifteen species and host twenty-two bacterial species belonging to seven classes in two different marine areas at varying rates were detected. The most common sponge-associated bacterium was recorded as Sphingomonas paucimobilis and Bacillus cereus in the Aegean Sea and the Sea of Marmara, respectively. The composition and counts of the sponge-associated bacteria were found significantly higher than the free-living bacteria in the ambient sea water sampling points of both two marine areas. The presence of high antibacterial potential of sponge-related bacteria obtained in this study provided data for further studies on marine-derived antimicrobial agents, including the effects of environmental differences.