Low-head barriers are used to block adult sea lamprey (Petromyzon marinus) from upstream spawning habitat. However, these barriers may impact stream fish communities through restriction of fish movement and habitat alteration. During the summer of 1996, the fish community and habitat conditions in twenty-four stream pairs were sampled across the Great Lakes basin. Seven of these stream pairs were re-sampled in 1997. Each pair consisted of a barrier stream with a low-head barrier and a reference stream without a low-head barrier. On average, barrier streams were significantly deeper (df = 179, P = 0.0018) and wider (df = 179, P = 0.0236) than reference streams, but temperature and substrate were similar (df = 183, P = 0.9027; df = 179, P = 0.999). Barrier streams contained approximately four more fish species on average than reference streams. However, streams with low-head barriers showed a greater upstream decline in species richness compared to reference streams with a net loss of 2.4 species. Barrier streams also showed a peak in richness directly downstream of the barriers, indicating that these barriers block fish movement upstream. Using S??renson's similarity index (based on presence/absence), a comparison of fish community assemblages above and below low-head barriers was not significantly different than upstream and downstream sites on reference streams (n = 96, P > 0.05), implying they have relatively little effect on overall fish assemblage composition. Differences in the frequency of occurrence and abundance between barrier and reference streams was apparent for some species, suggesting their sensitivity to barriers.