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Increasing information supported that achieving high-rate mainstream deammonification through two-stage partial nitritation (PN)-anammox process should be a better option than through single-stage process. However, direct experimental evidence was limited so far. Herein, a two-stage PN-anammox process was successfully operated for nitrogen removal from low-strength wastewater in winter. Influent shift from synthetic wastewater to actual anaerobically pretreated sewage had little impact on the process performance. Promising nitrogen removal rates (NRRs) of 0.28-0.07 kg N m−3 d−1 with an average effluent concentration of 5.2 mg TN L−1 were achieved for the anaerobically pretreated sewage treatment at 15-7 °C. Moreover, nearly all the degradable COD in the pretreated sewage was steadily removed in the first-stage PN reactor, despite the varied influent COD concentrations of 22-78 mg L−1 and the operating temperature decrease, suggesting the positive role of the first-stage PN in protecting anammox bacteria. The low temperature seemingly was the only deterministic factor inhibiting the anammox activity, and hence made the anammox reaction to be the rate-limiting step for nitrogen removal in the two-stage PN-anammox process. Unexpectedly, nearly all the anammox bacteria remained active at low temperatures with the process actual anammox activity reached about 76-85% of their maximum potential, implying that higher NRRs would be easily realized through bioaugmentation or enrichment of anammox bacteria. Overall, the present investigation provides direct and valuable information for implementing the two-stage PN-anammox process to treat mainstream municipal wastewater. A control strategy was also proposed to optimize the operation of the two-stage mainstream deammonification process. [ABSTRACT FROM AUTHOR]