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Nitrous oxide (N2O) emissions from wastewater treatment contribute significantly to greenhouse gas emissions. They have been shown to exhibit a strong seasonal and daily profile in previously conducted monitoring campaigns. However, only two year-long online monitoring campaigns have been published to date. Based on three monitoring campaigns on three full-scale wastewater treatment plants (WWTPs) with different activated sludge configurations, each of which lasted at least one year, we propose a refined monitoring strategy for long-term emission monitoring with multiple flux chambers on open tanks. Our monitoring campaigns confirm that the N2O emissions exhibited a strong seasonal profile and were substantial on all three plants (1–2.4% of the total nitrogen load). These results confirm that N2O is the most important greenhouse gas emission from wastewater treatment. The temporal variation was more distinct than the spatial variation within aeration tanks. Nevertheless, multiple monitoring spots along a single lane are crucial to assess representative emission factors in flow-through systems. Sequencing batch reactor systems were shown to exhibit comparable emissions within one reactor but significant variation between parallel reactors. The results indicate that considerable emission differences between lanes are to be expected in cases of inhomogeneous loading and discontinuous feeding. For example, N2O emission could be shown to depend on the amount of treated reject water: lanes without emitted