The European corn borer (ECB), Ostrinia nubilalis, is the principal pest of maize in Central and South Europe. It is known to promote Fusarium verticillioides infection in maize grain, a recognized producer of fumonisins. Field experiments were performed in 2006 and 2007 in two sites in NW Italy to determine the effects of the timing of insecticide application on ECB damage, fungal ear rot and fumonisin contamination under natural conditions. Different insecticide application timings were compared, from maize flowering to approximately 15 days after the flight peak of adult ECB. At harvest, the ears were rated for incidence and severity of ECB damage, fungal ear rot symptoms and fumonisin (FB(1) + FB(2)) contamination. In all years/sites, treatments applied at the beginning of consistent ECB flight activity were most effective in controlling insect damage on ears. Fungal ear rot and fumonisin contamination were significantly affected by ECB control. The efficacy of the best timing of insecticide application in controlling fumonisin contamination was, on average, 93% compared to the untreated control. Contamination levels of these mycotoxins increased with either an earlier or later treatment. Furthermore, an earlier insecticide application showed lower fumonisin contamination than a treatment applied after the adult flight peak. Production of maize kernels and maize-based foods that do not exceed the maximum international and EU permitted levels for fumonisins could be enhanced by appropriate insecticide treatment against second generation ECB. The optimum time for insecticide application is between the beginning of consistent adult flight activity and the flight peak.