Five study sites, and a sampling reach within each site, were established on the Rio Grande in and near Big Bend National Park in 1999 to provide the National Park Service with data and information on the status of stream habitat, fish communities, and benthic macroinvertebrates. Differences in stream-habitat conditions and riparian vegetation reflect differences in surface geology among the five sampling reaches. In the most upstream reach, Colorado Canyon, where igneous rock predominates, streambed material is larger; and riparian vegetation is less diverse and not as dense as in the four other, mostly limestone reaches. Eighteen species of fish and a total of 474 individuals were collected among the five reaches; 348 of the 474 were minnows. The most fish species (15) were collected at the Santa Elena reach and the fewest species (9) at the Colorado Canyon and Johnson Ranch reaches. The fish community at Colorado Canyon was least like the fish communities at the four other reaches. Fish trophic structure reflected fish-community structure among the five reaches. Invertivores made up at least 60 percent of the trophic structure at all reaches except Colorado Canyon. Piscivores dominated the trophic structure at Colorado Canyon. At the four other reaches, piscivores were the smallest trophic group. Eighty percent of the benthic macroinvertebrate taxa collected were aquatic insects. Two species of blackfly were the most frequently collected invertebrate taxon. Net-spinning caddisflies were common at all reaches except Santa Elena. The aquatic-insect community at the Boquillas reach was least similar to the aquatic-insect community at the other reaches.