Vannella simplex (Gymnamoebia, Vannellidae) is one of the most common amoebae species, recorded from a variety of regions. It was originally described as a freshwater species, but has also been reported from shallow-water regions of the Baltic Sea. In the present work, we investigated the morphology and biology of three V. simplex isolates, originating from geographically distant regions. Among them is one brackish water strain, isolated from artificial cyanobacterial mats, which were originally sampled in Nivå Bay (Baltic Sea, The Sound). The strain is cyst-forming and can thrive at salinity ranges from 0–50 ppt. Phylogenetic relationships were investigated by sequencing partial SSU rDNA of the cultured V. simplex isolates. Additional sequences were obtained from four environmental DNA extractions of sediment samples collected from different localities in Switzerland. Analysis of all obtained sequences revealed a monophyletic group. Based on the analysis and comparison of morphological, ecological and molecular data sets we compiled a distribution map of V. simplex and propose an emendation of this species.