Mercury (Hg) bioaccumulation in salamanders has received little attention despite widespread Hg contamination of aquatic ecosystems and worldwide amphibian declines. Here we report concentrations of methyl Hg (MeHg) and total Hg in larval northern two-lined salamanders (Eurycea bislineata bislineata) collected from streams in Acadia National Park (ANP), Maine, and Bear Brook Watershed, Maine (BBWM; a paired, gauged watershed treated with bimonthly applications (25 kg/ha/yr) of ammonium sulfate [(NH₄)₂SO₄]) since 1989), and Shenandoah National Park (SNP), Virginia. MeHg comprised 73–97% of total Hg in the larval salamander composite samples from ANP. At BBWM we detected significantly higher total Hg levels in larvae from the (NH₄)₂SO₄ treatment watershed. At ANP total Hg concentrations in salamander larvae were significantly higher from streams in unburned watersheds in contrast with larval samples collected from streams located in watersheds burned by the 1947 Bar Harbor fire. Additionally, total Hg levels were significantly higher in salamander larvae collected at ANP in contrast with SNP. Our results suggest that watershed-scale attributes including fire history, whole-catchment (NH₄)₂SO₄ additions, wetland extent, and forest cover type influence mercury bioaccumulation in salamanders inhabiting lotic environments. We also discuss the use of this species as an indicator of Hg bioaccumulation in stream ecosystems.