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The competition-colonization trade-off has long been considered an important mechanism explaining species coexistence in spatially structured environments, yet data supporting it remain ambiguous. Most competition-colonization research examines plants and the dispersal-linked traits of their seeds. However, colonization is more than just dispersal because rapid population growth is also an important component of colonization. We tested for the presence of competition-colonization trade-offs with a commonly used artificial assemblage consisting of protozoan and rotifer species, where colonization was the ability of a species to establish populations in patches. By ranking species according to their colonization abilities and their pairwise competitive interactions, we show that these species exhibit competition-colonization trade-offs. These results reveal that the competition-colonization trade-off exists within nonplant assemblages and that even in a laboratory setting, species are constrained to be either good competitors or colonizers but not both.