Molecular evidence for the involvement of Anopheles nivipes (Diptera: Culicidae) in the transmission of Plasmodium falciparum in north-eastern India.

The Anopheles philippinensis and An. nivipes mosquitoes that form part of the An. annularis species group are morphologically very similar and difficult to differentiate as adults. In consequence, researchers generally refer to all field-collected individuals of these species simply as the An. philippinensis-nivipes species complex. Although this species complex is understood to play a role in the transmission of parasites causing human malaria in north-eastern India, the identity of the exact species involved, and the relative importance of the local An. philippinensis and An. nivipes, are far from clear. To settle this issue, house-frequenting female adults of the An. philippinensis-nivipes species complex were collected from 23 localities in the six north-eastern states, identified to species (using an allele-specific PCR to explore the insects' ribosomal-DNA internal-transcribed-spacer-2 sequences) and checked for Plasmodium DNA (using a nested PCR based on the 18S subunit of the parasite's ribosomal DNA). Of the 337 females of the An. philippinensis-nivipes species complex that were investigated, 275 were identified as An. nivipes and 62 as An. philippinensis. Malarial infection was detected in the heads/thoraces of just two specimens, with P. falciparum DNA detected in one An. nivipes from Nagaland state and one An. nivipes from Assam state. These results provide unambiguous evidence of the presence of both An. philippinensis and An. nivipes in the north-eastern region of India and the involvement of An. nivipes in transmitting P. falciparum in this area.