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Background   Anopheles cruzii  is the primary human  Plasmodium  vector in southern and southeastern Brazil. The distribution of this mosquito follows the coast of the Brazilian Atlantic Forest. Previous studies indicated that  An. cruzii  is a complex of cryptic species.    Results  A multilocus approach using six loci, three circadian clock genes and three encoding ribosomal proteins, was implemented to investigate in more detail the genetic differentiation between the  An. cruzii  populations from Santa Catarina (southern Brazil) and Bahia States (northeastern Brazil) that represent two sibling species. The analysis revealed very high  F   ST   values and fixed differences between the two  An. cruzii  sibling species in all loci, irrespective of their function. An Isolation with Migration model was fit to the data using the IM program. The results reveal no migration in either direction and allowed a rough estimate of the divergence time between the two sibling species.    Conclusions  Population genetics analysis of  An. cruzii  samples from two Brazilian localities using a multilocus approach confirmed that they represent two different sibling species in this complex. The results suggest that the two species have not exchanged migrants since their separation and that they possibly diverged between 1.1 and 3.6 million years ago, a period of intense climatic changes.

Estimation of divergence time between two sibling species of the Anopheles (Kerteszia) cruziicomplex using a multilocus approach