The aim of this study was to investigate the suitability of a novel electrospun polyurethane nanofibre material for water-treatment purposes. Bacterial removal efficiency was tested in the laboratory by filtering artificial water spiked with Escherichia coli through a 0.25 µm nanofibre membrane. The results were compared with those obtained using a commercial microfiltration material (MV020T) with a similar pore size (0.20 µm). Alongside the laboratory experiments, we also determined filtration efficiency with semi-pilot scale experiments using actual wastewater from the secondary sedimentation tank of a wastewater treatment plant. The laboratory experiments indicated very high log10 removal efficiency, ranging from 5.8 to 6.8 CFU (colony-forming units)/ml. These results were better than those of the commercial membrane (3.8–4.6 CFU/ml). The semi-pilot scale experiment confirmed the membrane's suitability for microbial filtration, with both E. coli and total culturable microorganisms (cultured at both 22 and 36 °C) showing a significant decline compared to the non-filtered control (wastewater from the secondary outlet).