The Shenandoah Watershed Study (established in 1979) and the Virginia Trout Stream Sensitivity Study (established in 1987) serve to increase understanding of hydrological and biogeochemical changes in western Virginia mountain streams that occur in response to acidic deposition and other ecosystem stressors. The SWAS-VTSSS program has evolved over its 40+ year history to consist of a temporally robust and spatially stratified monitoring framework. Currently stream water is sampled bi-hourly during high-flow events at 3 sites and weekly at 4 sites within Shenandoah National Park (SHEN), and quarterly at 72 sites and on an approximately decadal frequency at ~ 450 sites within the wider western Virginia Appalachian region. Stream water is evaluated for pH, acid neutralizing capacity (ANC), base cations (calcium, magnesium, sodium and potassium ion), acid anions (sulfate, nitrate and chloride), silica and ammonium, with a subset of samples evaluated for monomeric aluminum and dissolved organic carbon. Hourly stream discharge (4 sites) and in-situ measurements of conductivity, water and air temperature (3 sites) are also measured within SHEN. Here we provide an overview and timeline of the SWAS-VTSSS stream water monitoring program, summarize the field and laboratory methods, describe the water chemistry and hydrologic data sets, and document major watershed disturbances that have occurred during the program history. Website links and instructions are provided to access the stream chemistry and time-series monitoring data in open-access federal databases. The purpose of this publication is to promote awareness of these unique, long-term data sets for wider use in catchment studies. The water chemistry and hydrologic data can be used to investigate a wide range of biogeochemical research questions and provide key inputs for models of these headwater stream ecosystems. SWAS-VTSSS is an ongoing program and quality assured data sets are uploaded to the databases annually.