Pseudomonas strains may be used as alternatives to fungicides as some of them produce secondary metabolites, which can inhibit growth of plant pathogenic fungi. Increased knowledge of non-target effects of the antagonistic bacteria on other soil organisms as well as of the survival and predation resistance of the antagonistic bacteria is necessary for risk assessment and increased performance of antagonistic bacteria as biological control agents. In the present study, we aimed to investigate the difference between Pseudomonas spp. with respect to their predation resistance to and effects on the three different and common soil flagellates Bodo caudatus, Cercomonas longicauda, and Neocercomonas jutlandica. Two antagonistic Pseudomonas: Pseudomonas fluorescens CHA0 and P. fluorescens DR54 and two positive control strains: P. fluorescens DSM 50090T and Pseudomonas chlororaphis ATCC 43928 were studied. CHA0, which is reported to produce a range of different antibiotic substances, was inhibitory to all the tested flagellates whereas DR54, which produces viscosinamide, only inhibited the growth rate of B. caudatus. Removal of spent medium from a CHA0 culture reduced the growth inhibition of C. longicauda, although the growth rate was still lowered compared with DSM 50090T. In contrast, removal of spent medium from a DR54 culture had minor effect on C. longicauda. The flagellate B. caudatus was far more sensitive towards the antagonistic Pseudomonas strains of the tested flagellates.