In 2009, we repeated a freshwater turtle survey first conducted in 1976 in the Lower Rio Grande Valley (LRGV) of Texas to determine whether the abundance of freshwater turtles in the LRGV has changed over the past three decades. We captured significantly fewer red-eared sliders (Trachemys scripta elegans) and Texas spiny softshells (Apalone spinifera emoryi) in 2 recently urbanized counties (Cameron and Hidalgo), and more red-eared sliders and Texas spiny softshells in a nonurbanized county (Willacy). Land-use changes, increased urbanization, and commercial turtle harvest are likely responsible for the decline of freshwater turtles in the LRGV.