Ixodes scapularis (Acari: Ixodidae) is the principal vector of Borrelia burgdorferi (the etiologic agent of Lyme disease) in the eastern and midwestern United States. Recent efforts have documented the first established population of I. scapularis in South Dakota, representing a western expansion of the known species distribution. Our goal was to describe the current distribution of I. scapularis in eastern South Dakota and to survey for the presence of B. burgdorferi in questing I. scapularis. We surveyed for the presence of adult and nymphal I. scapularis in seven counties within South Dakota, including 13 locales from 2016 to 2017. We then tested all I. scapularis, including those collected in 2015 from a previous study, for the presence of B. burgdorferi sensu stricto, via quantitative and conventional polymerase chain reaction. Here, we document the presence of I. scapularis in four new counties in South Dakota, and report the first instance of B. burgdorferi in a questing tick in South Dakota. Coupled with data from the Centers for Disease Control and Prevention and Companion Animal Parasite Council our data show that the risk of contracting Lyme disease in South Dakota is low, but existent and should be an important consideration with regard to public health, pets, and wildlife.