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Background  Members of  Anopheles maculipennis  complex are effective malaria vectors in Europe and the Caspian Sea region in northern Iran, where malaria has been re-introduced since 1994. The current study has been designed in order to provide further evidence on the status of species composition and to identify more accurately the members of the maculipennis complex in northern Iran.    Methods  The second internal transcribed spacer of ribosomal DNA (rDNA-ITS2) was sequenced in 28 out of 235 specimens that were collected in the five provinces of East Azerbayjan, Ardebil, Guilan, Mazandaran and Khorassan in Iran.    Results  The length of the ITS2 ranged from 283 to 302 bp with a GC content of 49.33 – 54.76%. No intra-specific variations were observed. Construction of phylogenetic tree based on the ITS2 sequence revealed that the six Iranian members of the  maculipennis  complex could be easily clustered into three groups: the  An. atroparvus – Anopheles labranchiae  group; the paraphyletic group of  An. maculipennis, An. messeae, An. persiensis;  and  An. sacharovi  as the third group.    Conclusion  Detection of three species of the  An. maculipennis  complex including  An. atroparvus, An. messae  and  An. labranchiae , as shown as new records in northern Iran, is somehow alarming. A better understanding of the epidemiology of malaria on both sides of the Caspian Sea may be provided by applying the molecular techniques to the correct identification of species complexes, to the detection of Plasmodium composition in Anopheles vectors and to the status of insecticide resistance by looking to related genes.

Molecular identification of Palearctic members of Anopheles maculipennis in northern Iran