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BackgroundAnopheles stephensi is considered an important malaria vector in Iran, Asia, and recently in the Horn of Africa. Recently, Ansteobp1 intron I sequence has been introduced a new molecular marker for identification of its biological forms including, mysorensis, intermediate and type, using insectary colony specimens.MethodsIn the current study, new marker ability in molecular identification of biological forms has been evaluated with An. stephensi specimens collected from Iran and Afghanistan malarious provinces. Following DNA extraction and PCR amplification, sequence analysis and constructed phylogenetic tree revealed that type and intermediate forms are distributed in Iran.ResultsThe specimens collected from Afghanistan identified as intermediate and mysorensis forms. Therefore, intermediate form is sympatric species in both countries. Based on the results of Ansteobp1 intron I sequences, An. stephensi could be suggested as new Anopheles complex species including An. stephensi sibling A (type form), An. stephensi sibling B (intermediate form) and An. stephensi sibling C (mysorensis form). This is the first report on the presence of An. stephensi biological forms in Afghanistan.ConclusionsIran is going to eliminate malaria transmission from the country, precise species identification, especially in complex species will be helpful in the prevention of malaria resurgence in the country, mainly because of common fauna of Anopheles species and through border malaria and population movement within Afghanistan, Pakistan, and Iran.

In recent years, the increase of head louse infestation in Iran (7.4%) and especially in West-Azerbaijan Province (248%) has raised the hypothesis of insecticide resistance development. There are different mechanisms of resistance to various groups of insecticides, and knockdown resistance (kdr) is a prominent mechanism of resistance to pyrethroids, an insecticide group which is used conventionally for pediculosis control. For detection of kdr-type well-known amino acid substitutions (M815I-T917I-L920F) and additional sodium channel mutations potentially associated with kdr resistance in head and body lice, louse populations were collected from West-Azerbaijan and Zanjan Provinces of Iran. Six novel mutations were found to be located in the IIS1-2 extracellular loop (H813P) and IIS5 (I927F, L928A, R929V, L930M, and L932M) of the α-subunit. Genotyping results showed that all specimens (100%) have at least one of these or the well-known mutations. Therefore, the presence of kdr-related and novel mutations in the sodium channel is likely to be the reason for the frequent use of pyrethroid insecticides due to treatment failure against lice. Further studies are now required to evaluate the prevalence of the kdr-like mutant allele for monitoring of insecticide resistance and the management of head and body lice in other provinces of the country.

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