Biosensors for in situ detection of pathogenic bacteria in liquid are developed using magnetostrictive particles (MSP) as the sensor platform. The sensing elements used are phage E2 against Salmonella typhimurium, monoclonal antibody against Listeria monocytogenes, polyclonal antibody against Escherichia coli, and polyclonal antibody against Staphylococcus aureus, respectively. These biosensors were characterized in cultures with different populations ranging from 5 × 101 to 5 × 108 cfu/mL. It is found that the MSP-based biosensors work well in water and have a rapid response with a response time in minutes, which makes the MSP-based sensors suitable for in situ and real-time detection of pathogenic bacteria in liquid. The experimental results show that all MSP-phage and MSP-antibody biosensors in size of 1.0 mm × 0.3 mm × 15 µm exhibit a detection limit better than 100 cfu/mL. Based on the Hill plot, it is concluded that each bacterial cell is bound onto the sensor surface through about four-to-five sites. When the cultures with low population (<106 cfu/mL) are tested, both MSP-phage and MSP-antibody sensors exhibit the similar response. However, the phage-MSP sensors exhibit a higher capability in the capture of target bacterial cell. Biotechnol. Bioeng. 2014;111: 2229–2238. © 2014 Wiley Periodicals, Inc.