The Mery-sur-Oise plant (France) has been using nanofiltration (NF) membranes (NF200) to produce safe drinking water since 1999. However, significant fouling has been occasionally observed according to seasonal conditions, even with various pre-treatments including conventional surface water treatment followed by ozonation, acid addition to pH 6.9, anti-scalant addition, and microfiltration (6mum). Pilot-scale filtration experiments were performed to determine the effects of natural organic matter (NOM) character and ozonation on NF membrane fouling under constant operating conditions. Two parallel pilot units were operated with sand-filtered water (SFW) and sand-filtered-ozonated water (SFOW) for 3-month periods corresponding to spring and fall seasons. To identify NF foulants, Fourier transform infrared spectroscopy, fluorescence excitation emission matrix, scanning electron microscope, energy-dispersive spectrophotometry, and HPSEC-UVA-DOC-fluorescence chromatography have been used. Even though the dissolved organic carbon (DOC) and ultraviolet (UVA) levels of spring samples were lower than those of winter season, these feed waters showed higher fouling presumably due to a higher hydrophilic fraction of NOM and the presence of microorganisms. In addition, for both seasons, ozonation increased the degree of fouling mainly by a change in NOM characteristics and by the promotion of bacterial cell growth conditions. The hydrophilic NOM is not expected to be easily rejected by the relatively hydrophilic and negatively charged NF200 membrane due to its non-charged (or oppositely charged) properties, indicating a high fouling potential by NOM associated with spring samples. The adhesion of bacteria and accumulation of microalgae on the membrane may be due to the role of extracellular biopolymers released by algae upon ozonation, promoting adhesion between microorganisms and the membrane surface. Protein- and polysaccharide-like substances were found as major foulants. The reason for the minor fouling by humic substances on membranes fed with SFOW during the spring season might be a loss of membrane surface charge due to screening by significant subsequent fouling on the base of the fouling layer of extracellular materials.