The U.S. Geological Survey is studying uranium and associated trace element bioaccumulation in aquatic invertebrates across a gradient of dissolved uranium concentrations in spring outflow pools and creeks in the Grand Canyon and adjacent watershed. This data release makes available data from sampling campaigns in April 2016, April 2017, and April 2019. Data collected include: (1) major ion, trace element and dissolved organic carbon in surface waters of spring outflow pools and streams; (2) concentrations of uranium and other trace elements in bed sediment, periphyton, and larva of aquatic insect taxa; (3) uranium and iron extraction data from a subset of bed sediment samples, and (4) X-ray diffraction data for bed sediment samples collected in 2019. This dataset includes samples from the corrected original data release (invertebrate data for 2016 were incorrectly reported as mass concentrations (mg/kg) in the original csv file), plus samples collected in 2019. NOTE: While previous versions are available from the author, all the records in previous versions can be found in version 4.0. Direct questions regarding data to the authors.