Fish exhibiting external lesions, incidences of intersex, and death have recently been observed in the Shenandoah and James River Basins. These basins are characterized by widespread agriculture (intensive in some areas), several major industrial discharges, numerous sewage treatment plant discharges, and urban, transportation, and residential growth that has increased rapidly in recent years. Nine locations in the Shenandoah River Basin, Virginia, and two in the James River Basin, Virginia, were selected for study in an attempt to identify chemicals that may have contributed to the declining fish health. Two passive sampling devices, semipermeable membrane devices (SPMDs) and polar organic chemical integrative samplers (POCIS), were deployed during the spring and early summer of 2007 to measure select organic contaminants to which fish may have been exposed. This study determined that concentrations of persistent hydrophobic contaminants, such as polycyclic aromatic hydrocarbons (<17,000 picograms per liter), legacy pesticides (<510 picograms per liter), and polychlorinated biphenyls (<1,600 picograms per liter) were generally low and indicative of a largely agricultural area. Chlorpyrifos, endosulfan, and lindane were the most commonly detected chlorinated pesticides. Atrazine, which was detected at concentrations much greater than other pesticides associated with agricultural use, ranged from <0.18 to 430 nanograms per liter during the deployment period. Few chemicals characteristic of wastewater treatment plant effluent or septic tank discharges were detected. The fragrance components, galaxolide, indole, and tonalide, were the predominant waste indicator chemicals detected. Caffeine, the caffeine metabolite 1,7-dimethylxanthine, the nicotine metabolite cotinine, and the prescription pharmaceuticals carbamazepine, venlafaxine, and trimethoprim were detected at several sites. Natural and synthetic hormones were detected at a few sites with 17α-ethynylestradiol concentrations esimated up to 8.1 nanograms per liter. Screening of the POCIS extracts for estrogenic chemicals by using the yeast estrogen screen revealed estrogenicity similar to levels reported for rural areas with minor effect from wastewater effluents.