Over a 22 month period Barbus graellsii, Chondrostoma toxostoma, Cyprinus carpio, Esox lucius, Gobio gobio, and Leuciscus cephalus displayed non-random microhabitat use in the Rio Mattarraña, Spain and generally were overrepresented in deep microhabitats with low or undetectable flow velocities. Substrate composition did not strongly affect microhabitat use outside of its covariation with depth and velocity. Most seasonal differences in microhabitat use were attributable to seasonal changes in microhabitat availability, although all species selectively occupied deeper microhabitats during Spring 1984, 1985, and Early Summer 1984. Smaller specimens of B. graellsii, Ch. toxostoma, G. gobio, and L. cephalus all occurred closer to the substrate than larger specimens. Smaller specimens also tended to occupy shallower areas with greater amounts of erosional substrates (except for Ch. toxostoma). Assemblage members occupied statistically distinct microhabitats and could be classified as: 1) upper water column (L. cephalus), 2) mid-water column (Ch. toxostoma, C. carpio), 3) lower water column (B. graellsii), or 4) benthic (G. gobio, E. lucius). We hypothesize that the observed pattern of vertical segregation was produced by a combination of predator avoidance and differential evolutionary adaptation rather than by interspecific competition for space.