Five malaria mosquitoes were identified in the Amudarya river valley. An. pulcherrimus, An. martinius, An. superpictus, and An. hyrcanus inhabit in the plain of the valley. An. superpictus, An. pulcherrimus, An. hyrcanus, and An. claviger occur in the head of the Amudarya river. The nucleotide composition of the second inner transcribed spacer (ITS2) of ribosomal DNA and the locus of cytochromoxidase I (COI) of mitochondrial DNA was determined in the closely related species An. martinius and An. artemievi. It is shown that the loci in question may be used to diagnose twin species of the maculipennis complex.

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