Study of Stenamoeba species from several soil habitats revealed two new species, named Stenamoeba berchidia and Stenamoeba sardiniensis. Both of them demonstrate morphology, characteristic to this genus and group in SSU rDNA trees with other members of the genus Stenamoeba, forming a highly supported clade within the family Thecamoebidae. The ultrastructural investigation of Stenamoeba sardiniensis revealed the presence of cytoplasmic MTOC, located close to one of several dictyosomes found inside the cell. This is the first report of cytoplasmic MTOC among Thecamoebidae; the presence of MTOCs is now shown in two of the three orders comprisin+g the subclass Longamoebia. We also report the isolation of Stenamoeba limacina from German soils. This strain shows a similar morphology and an almost complete SSU rDNA sequence identity with the type strain of S. limacina originating from gills of fishes, collected in Czech Republic.