This report documents and summarizes physical, chemical, and biological data collected during 1999-2004 in a study titled Effects of Urbanization on Stream Ecosystems, undertaken as part of the U.S. Geological Survey's National Water-Quality Assessment Program. Data-collection methods and data processing are described in this report for streamflow; stream temperature; instream chemistry; instream aquatic habitat; and algal, macroinvertebrate, and fish communities. Data summaries prepared for analytical use are presented in downloadable data tables.