The larvae of the genus Chironomus are a common object for hydrobiological studies, as well as a model object for cytogenetics. Morphologically, the species are very similar. One of these species or species complex is Chironomus “annularius”, which has a Holarctic distribution. It has chromosomal banding sequences characteristic of Nearctic and Palearctic populations. Using an integrated method that included morphology, cytogenetics, and molecular genetics, we analyzed populations from Russia, Mongolia, and Armenia. We found through cytogenetics and larval morphology that the populations have high similarity. Molecular genetic studies have shown significant differences between the populations. The genetic distances between the populations, in some cases, exceed the interspecific threshold of 3%, and are 6.5%. In the South Caucasian population (Lake Sevan), a chromosomal banding sequence, h’annD3, that was previously observed only in North America, was found for the first time. The larvae from Lake Sevan have large genetic distances from others, and are morphologically similar to the species Chironomus markosjani Shilova 1983, described from this lake without comparison with Ch. annularius nor an exact description of the karyotype. The sequences of the COI genes from Montenegro (Lake Skadar) and West Siberia (Novosibirsk) found in GenBank may belong to a new undescribed species, or a species not represented in the database. Thus, the analyzed data on Chironomus “annularius” support the presence of the complex of homosequential species under this name.