Molecular characteristics of Anopheles maculipennis Meigen in Zanjan, north west of Iran, inferred from ITS2 sequence analysis.

The present study has been designed in order to verify the species composition within Anopheles maculipennis complex in North West of Iran. We determined ribosomal DNA sequences of the second internal transcribed spacer (ITS2) region from samples of Anopheles maculipennis Complex in Zanjan province. A total of 1536 specimens within the Complex were tested by Multiplex PCR, only An. maculipennis was found in this area. One clone out of four different individual mosquitoes of each field was generated with ITS2 PCR and half of them (192 samples) selected randomly for RFLPs. PCR-RFLP assay identified 2 haplotypes; haplotype I (99%) and haplotype II (1%). Twenty five sequences were generated comprising the 5.8S gene, the ITS2 and the 28S ribosomal gene. The alignment was 422 in length and percentage of GC content was 50.3% (26.07% A, 23.59% T, 26.78% C, 23.7% G). The ITS2 was 290 bp in length and two haplotypes were revealed varying by a single base (T<-->C) at site 378. An. maculipennis is the dominant species anopheline of the province. ITS2 analysis revealed evidence of a slightly interaspecific variation among populations. However, further investigations on the genetic polymorphism among An. maculipennis populations and in particular within those belonging to the continental haplotype are required to support any hypothesis on differences in behavior across the distribution range for this potential malaria vector.