The antibacterial behaviour of suspensions of zinc oxide nanoparticles (ZnO nanofluids) against E. Coli has been investigated. ZnO nanoparticles from two sources are used to formulate nanofluids. The effects of particle size, concentration and the use of dispersants on the antibacterial behaviour are examined. The results show that the ZnO nanofluids have bacteriostatic activity against E. coli. The antibacterial activity increases with increasing nanoparticle concentration and increases with decreasing particle size. Particle concentration is observed to be more important than particle size under the conditions of this work. The results also show that the use of two types of dispersants (Polyethylene Glycol (PEG) and Polyvinylpyrolidone (PVP)) does not affect much the antibacterial activity of ZnO nanofluids but enhances the stability of the suspensions. SEM analyses of the bacteria before and after treatment with ZnO nanofluids show that the presence of ZnO nanoparticles damages the membrane wall of the bacteria. Electrochemical measurements using a model DOPC monolayer suggest some direct interaction between ZnO nanoparticles and the bacteria membrane at high ZnO concentrations.