The U.S. Geological Survey, in collaboration with the Houlton Band of Maliseet Indians (HBMI), conducted a pilot microbial source tracking (MST) study to investigate sources of fecal indicator bacteria in the Meduxnekeag River watershed near Houlton, Maine. HBMI staff collected surface water samples from three sites in June 2026. Samples were analyzed using quantitative polymerase chain reaction (qPCR) assays targeting human- and canine-associated MST markers on July 14, 2026. This data release contains the environmental sample results and associated quality assurance and quality control (QA/QC) data.