Two polyvinylidene microfiltration (MF) membranes with a nominal pore size of 0.22 μm were challenged with mixed microbial cultures present in Milli-QTM water and in secondary effluent, and with a Gram-negative model bacterium, SW8, to investigate bacterial passage. Total bacterial counts measured microscopically using the DNA fluorochrome DAPI revealed that the small bacteria in Milli-QTM water passed MF membranes totally. SW8 and bacteria from secondary effluent were mostly retained with log reduction values of 4 and 3.5, respectively. Transmembrane pressure did not influence the levels of bacterial passage significantly. Pore size effects were investigated with tracked-etched membranes with nominal pore sizes of 0.2, 0.1 and 0.05 μm.