We quantified the influence of agricultural activities on environmental and biological conditions of floodplain wetlands in the upper Missouri River basin. Seasonally-flooded wetlands were characterized as low impact (non-disturbed) or high impact (disturbed) based on local land use. Biological data collected from these wetlands were used to develop a wetland condition index (WCI). Fourteen additional wetlands were sampled to evaluate the general condition of seasonally-flooded floodplain wetlands. Structural and functional attributes of macrophyte, algae, and macroinvertebrate communities were tested as candidate metrics for assessing biotic responses. The WCI we developed used six biological metrics to discriminate between disturbed and non-disturbed wetlands: 1) biomass of Culicidae larvae, 2) abundance of Chironomidae larvae, 3) macroinvertebrate diversity, 4) total number of plant species, 5) the proportion of exotic plant species, and 6) total number of sensitive diatom species. Disturbed wetlands had less taxa richness and species diversity and more exotic and nuisance (e.g., mosquitoes) species. Environmental differences between low and high impact wetlands included measures of total potassium, total phosphorus, total nitrogen, alkalinity, conductance, and sediment phosphorus concentration. Canonical analyses showed that WCI scores were weakly correlated (P = 0.057) with environmental variables in randomly selected wetlands. In addition, mean WCI score for random wetlands was higher than that for high impact wetlands, implying that floodplain wetlands were less impacted by the types of agricultural activities affecting high impact sites. Inter-year sampling of some wetlands revealed that WCI metrics were correlated in 2000 and 2001, implying that biological metrics provided useful indicators of disturbance in floodplain wetlands.