Ultrasound technology has attracted increasing attention in the field of sewage sludge treatment. This study investigated the nitrite accumulation ratio (NAR) and microbial characteristics of the partial nitrification (PN) process in a sequencing batch reactor employing ultrasonic treatment (ultrasound density = 0.25 W/mL, irradiation time = 10 min). PN was achieved over 73 days with a NAR above 85% under ambient temperatures. A low dissolved oxygen (DO) environment was generated in the reactor by enhancing the oxygen utilization rate of ammonia-oxidizing bacteria (AOB). Additionally, the application of long-term ultrasonic treatment led to the enhancement of the dominance of the Nitrosomonas genus of AOB, while populations of the Nitrospira genus of nitrite-oxidizing bacteria (NOB) were eradicated. At the same time, the activities of the aerobic denitrifying bacteria Thauera, Terrimonas, Defluviimonas, and Thermomonas were enhanced and their relative abundance was increased. Overall, the results suggest that ultrasonic treatment can enhance AOB activity and generate a low DO environment that facilitates effective PN.