Advanced membranes that enable ultrafast water flux while demonstrating anti-biofouling characteristics can facilitate sustainable water/wastewater treatment processes. MXenes, two-dimensional (2D) metal carbides and nitrides, have attracted attention for applications in water/wastewater treatment. In this work, we reported the antibacterial properties of micrometer-thick titanium carbide (Ti3C2Tx) MXene membranes prepared by filtration on a polyvinylidene fluoride (PVDF) support. The bactericidal properties of Ti3C2Tx modified membranes were tested against Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis) by bacterial growth on the membrane surface and its exposure to bacterial suspensions. The antibacterial rate of fresh Ti3C2Tx MXene membranes reaches more than 73% against B. subtilis and 67% against E. coli as compared with that of control PVDF, while aged Ti3C2Tx membrane showed over 99% growth inhibition of both bacteria under same conditions. Flow cytometry showed about 70% population of dead and compromised cells after 24 h of exposure of both bacterial strains. The damage of the cell surfaces was also revealed by scanning electron microscopy (SEM) and atomic force microscopy (AFM) analysis, respectively. The demonstrated antibacterial activity of MXene coated membranes against common waterborne bacteria, promotes their potential application as anti-biofouling membrane in water and wastewater treatment processes.