A novel Gram-negative, obligate aerobic, mobile, rod-shaped and non-spore-forming bacterial strain, WCHP16T, was isolated from the wastewater treatment plant at West China Hospital in Chengdu, PR China. It was characterized using a polyphasic approach. Analysis of its 16S rRNA gene sequence showed that strain WCHP16T belonged to the genus Pseudomonas with the highest similarity to Pseudomonas qingdaonensis JJ3T (99.34 %), Pseudomonas shirazica VM14T (99.0 %), Pseudomonas plecoglossicida NBRC 103162T (99.0 %) and Pseudomonas asiatica RYU5T (99.0 %). Phylogenomic analysis based on 107 core gene sequences demonstrated that WCHP16T was a member of the Pseudomonas putida group but was distant from all closely related species. Whole-genome comparisons, using average nucleotide identity based on blast (ANIb) and in silico DNA–DNA hybridization (isDDH), confirmed low genome relatedness to all the known Pseudomonas species (below the recommended thresholds of 95 % [ANIb] and 70 % [isDDH] for species delineation). Phenotypic characterization tests showed that the utilization of phenylacetic acid and capric acid, but not d-arabitol, and inability to produce fluorescent (King B medium) in combination could distinguish this strain from other related species of the genus Pseudomonas . Therefore, based on genetic and phenotypic evidence, it is clear that strain WCHP16T represents a novel species, for which the name Pseudomonas defluvii sp. nov. is proposed. The type strain is WCHP16T (GDMCC1.1215T=CCTCC AB 2017103T=KCTC 52991T).