Hydrilla [Hydrilla verticillata (L. f.) Royle], an invasive aquatic weed, has had a rich introduction history into the United States, with multiple subspecies being introduced since the 1960s. The most recent occurred before 2016, when northern hydrilla (Hydrilla verticillata ssp. lithuanica) was discovered in the Connecticut River. By 2021, following a 3-yr survey from Agawam, MA, to the Long Island Sound by the Connecticut Agricultural Experiment Station Office of Aquatic Invasive Species, H. verticillata ssp. lithuanica was found in more than 113 km of the river, occupying 344 ha. Since this survey, there has been concern that H. verticillata ssp. lithuanica would spread to nearby waterbodies and have a significant negative impact. Here, we report the first documented spread and establishment of H. verticillata ssp. lithuanica from the Connecticut River to five waterbodies in Connecticut and one in Massachusetts. Of the eight sites where H. verticillata observations were made, 75% (n = 6) were confirmed to be H. verticillata ssp. lithuanica and 25% (n = 2) to be Hydrilla verticillata ssp. peregrina (wandering hydrilla). Except for one site, all six locations infested with H. verticillata ssp. lithuanica provide watercraft access through public or private boat ramps. The authors also postulate on the mechanisms facilitating the spread and establishment of this subspecies.