Display omitted •Achieving PN in domestic wastewater controlled by aeration time.•AOB and NOB activities eliminated entirely during simultaneous elimination period.•Prolonged aeration time led to AOB quickly recovery, on the contrary to NOB.•Stable PN maintained for 205 days with high NAR (96%) and ARE (94.3%).•Without additional chemical agents and infrastructure costs for this strategy. This study developed a novel strategy for rapidly achieving partial nitrification (PN) without additional chemical agents, and infrastructure costs, only by controlling aeration time to selectively enrich ammonium oxidizing bacteria (AOB) after simultaneously eliminating AOB and nitrite oxidizing bacteria (NOB). Shorter aeration time and sludge retention time (10 days) were implemented to simultaneously eliminate AOB and NOB, the bioactivities drastically decreased to 13 and 0%, respectively. Subsequently, a gradually prolonged aeration time selectively enriched AOB and resulted in PN. The amoA abundances increased to 1.9 × 1010 copies gVSS−1, whereas Nitrospira and Nitrobacter abundances remained stable (3.2 × 109 and 3.1 × 109 copies gVSS−1). A nitrite accumulation rate above 96% was achieved and maintained for 205 days over the entire temperature range (28.5–17.9 °C). The effluent contained 1.9 mg N L−1 of ammonium, 25.3 mg N L−1 of nitrite, and less than 1.0 mg N L−1 of nitrate, facilitating mainstream wastewater anammox.