Recent genetic evaluations of R. lucifugus from France and Italy with R. lucifugus from Turkey suggests that these geographic groups likely constitute separate species. A more extensive review of eastern Mediterranean termites is needed to clarify their respective classifications. DNA sequencing analysis of the mitochondrial 16S rRNA region was used to examine genetic variation in the termite genus Reticulitermes Holmgren from eastern Europe and the Middle East. Sequencing of a 438 bp region from 39 individuals and from 3 sequences obtained from GenBank revealed 47 polymorphic sites within the genus. Tajima-Nei distances from species ranged from 0.0 to 4.9%, and parsimony and maximum likelihood analysis revealed clades within the genus which follow biogeographic patterns of dispersal that may support recent glacial refugia theories. European Reticulitermes were delimited from Turkish R. lucifugus with samples from Greece, Israel, and Iran. Reticulitermes lucifugus from Turkey share a topology with R. clypeatus from Israel and R. sp. from Iran, which were distinct relative to other Reticulitermes from Eastern Europe. There was some genetic support for combining R. balkanensis to this group. However, this study suggests that Reticulitermes from this region likely constitute two genetically isolated, but sympatric populations which have not been resolved with morphological examinations, but are clearly different from the R. lucifugus complex from Europe.