Water quality in the Newfound Creek watershed has been shown to be affected by bacteria, sediment, and nutrients. In this study, Escherichia coli (E. coli) bacteria were sampled at five sites in Newfound Creek and five tributary sites during low flow on May 28, 2003, and high flow on November 19, 2003. In addition, a subset of five sites was sampled for fecal coliform bacteria, E. coli bacteria in streambed sediments (low flow only), and coliphage virus for serotyping. Coliphage virus serotyping has been used to identify human and animal sources of bacterial contamination. A streamflow gage was installed and operated to support ongoing water-quality studies in the watershed. Fecal coliform densities ranged from 92 to 27,000 colony-forming units per 100 milliliters of water for E. coli and 140 to an estimated 29,000 colony-forming units per 100 milliliters of water for fecal coliform during the two sampling visits. Ninety percent of the E. coli and fecal coliform samples exceeded corresponding U.S. Environmental Protection Agency or North Carolina water-quality criteria for recreational and ambient waters. During low flow, the middle part of the Newfound Creek watershed and the Dix Creek tributary had the highest densities of E. coli bacteria. During the high-flow sampling, all tributaries contained high densities of E. coli bacteria, although Dix Creek and Round Hill Branch were the largest contributors of these bacteria to Newfound Creek. Coliphage virus serotyping results were inconclusive because most samples did not contain the male-specific RNA coliphage needed for serotyping. Positive results indicated, however, that during low flow, non-human sources of bacteria were present in Sluder Branch, and during high flow, human sources of bacteria were present in Round Hill Branch. Sampling of bacteria in streambed sediments during low flow indicated that sediments do not appear to be a substantial source of bacteria relative to the water column, with the exception of an area near the confluence of Sluder Branch and Newfound Creek.