Like most streams located in the Ozark Plateaus, the Buffalo River in Arkansas generally has excellent water quality. Water-quality conditions in Big Creek, however, a major tributary of the middle Buffalo River, have been less favorable than that of other Buffalo River tributaries. Concerns regarding the influence of water quality in Big Creek on the Buffalo River magnified in 2013 when a large confined animal feeding operation (CAFO) began operating in the watershed. In response to these concerns, the U.S. Geological Survey compared monthly nutrient concentrations and seasonal periphyton assemblage metrics of a site on Big Creek downstream of the CAFO, two Buffalo River control sites upstream of the confluence with Big Creek, and three Buffalo River test sites downstream of the confluence with Big Creek. In addition to identifying potential nutrient patterns and periphyton responses along a low-level nutrient exposure gradient, the study determined how nutrient contributions from Big Creek (and the CAFO) are affecting ecological conditions and consequent ecosystem services in the Buffalo River. Nutrient and periphyton data exhibited more temporal than spatial variability. Nutrient concentrations were generally highest of all sites at the Big Creek site. Concentrations at the five sites on the Buffalo River were typically low (near laboratory reporting limits), and concentrations at the three test sites rarely exceeded those of the two control sites. An index developed with three ecologically relevant periphyton metrics (oligotrophic taxa and Homoeothrix percent relative abundance and mesotrophic diatoms percent taxa richness) suggested that nutrient uptake at sites downstream of the Big Creek-Buffalo River confluence resulted in subtle shifts in downstream periphyton assemblages. The periphyton index of biological integrity at control sites was slightly and generally more favorable compared to test sites. Even so, when periphyton data were considered in conjunction with both hydrology and water-quality data, the negative consequences of antecedent high flows and associated scouring exceeded the potential positive effects that low-level nutrients had on algal productivity. These findings emphasize the importance of comparing biological and chemical data across extended temporal scales, particularly when working with low-level nutrient gradients.