In total, 289 termite samples were collected from 45 Counties in Indiana during 2002-2004. Approximately 89% of the collection sites were associated with artificial structures, and almost half of the samples were from inside homes. The rest of the samples were from forested areas. Termite samples were identified based on their morphological characteristics, molecular characteristics, or both. Five species from the genus Reticulitermes were identified, and the relative abundance (percentage of the total collections) of these five species was Reticulitermes flavipes (Kollar) (90.0), Reticulitermes virginicus (Banks) (7.6), Reticulitermes arenincola (Goellner) (1.0), Reticulitermes tibialis (Banks) (1.0), and Reticulitermes hageni Banks (0.3). Based on the distribution map, R.flavipes was the dominant and the most widely distributed species in Indiana (44 counties); followed by R. virginicus (13 counties). The three other Reticulitermes species, R. arenincola, R. tibialis, and R. hageni, were encountered in only five counties. R. arenincola is considered a rare species and its distribution has been limited to sand dunes near Lake Michigan. However, in this study, two of the three R. arenincola samples were collected outside of its type location. R. tibialis was found in three counties, whereas R. hageni was only found in Evansville, IN. To complement the morphological identifications, a 389-bp region of the mitochondrial DNA (mtDNA) 16S rRNA gene was amplified and sequenced from all five Reticulitermes species. Based on species-specific polymorphisms exhibited in mtDNA sequences, a polymerase chain reaction-restriction fragment length polymorphism-based diagnostic tool was developed to identify samples lacking of diagnostic morphological characters.