A Gram-negative, rod-shaped, non-spore-forming, yellow-pigmented bacterium (strain B2T) isolated from wastewater of a sequence batch reactor showing enhanced phosphorus removal was investigated to determine its taxonomic status. Complete 16S rRNA gene sequence analysis indicated that the organism should be placed in the genus Chryseobacterium. The strain contained a polyamine pattern with sym-homospermidine as the major compound, menaquinone MK-6 as the predominant menaquinone and ai-C15:0, i-C15:0 and C16:1 as the major fatty acids. Phosphatidylethanolamine and several unidentified lipids were detected in the polar lipid profile. Phylogenetically, strain B2T was most closely related to Chryseobacterium indoltheticum and Chryseobacterium gleum (96.2 and 95-9% 16S rRNA gene sequence similarity, respectively). The phylogenetic distance from any validly described species within the genus Chryseobacterium, as indicated from 16S rRNA gene sequence similarities, and its phenotypic properties demonstrate that strain B2T represents a novel species, for which the name Chryseobacterium defluvii sp. nov. is proposed the type strain is B2T (=DSM 14219T =CIP 107207T).