Abstract • Pseudomonas mendocina was first reported for aerobic nitrate removal. • It removed 90% of NO3−-N in 24 h under aerobic conditions. • This strain converted NO3−-N to bio-nitrogen (37.9%) and gaseous nitrogen (49.7%). • Inoculation of this strain increased sludge denitrification rate by 4.3 times. The problem of nitrate accumulation in aerobic tank and total nitrogen excessive discharge in effluent was very common in traditional livestock and poultry farming wastewater treatment systems owing to the lengthy process flow and low process control level. A strain LYX of aerobic bacterium was isolated from the activated sludge of a wastewater treatment system in a pig farm, which could remove nitrate effectively in aerobic tank and was identified Pseudomonas mendocina by 16S rRNA sequencing. Under the condition of nitrate as the sole nitrogen source, this strain removed over 90% of NO3−-N with an initial concentration of 110 mg/L under aerobic conditions within 48 hours. Among them, 37.9% of NO3−-N was assimilated into Bio-N, about 51.9% was reduced to gaseous nitrogen and less than 0.5% of nitrogen was replaced by NO2−-N and NH4+-N, 9.7% NO3−-N remained in the effluent at the end. At the same time, four key genes (napA, nirK, norB and nosZ) related to nitrate nitrogen removal were expressed during the denitrification process of P. mendocina LYX, in which the transcription level of the indicator genes of this aerobic denitrifying bacterium (napA) was the highest. In addition, it was found with the 15N tracer technique that inoculation of this strain on sludge increased the amount of nitrogen loss from 9.26 nmol N/(g·h) to 23.835 nmol N/(g·h). Therefore, P. medocina LYX is a potential bioagent for advanced nitrogen removal by assimilating and reducing nitrate simultaneously in aerobic tanks.