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Background   Anopheles  ( Kerteszia )  cruzii  is a primary vector of  Plasmodium  parasites in Brazil’s Atlantic Forest. Adult females of  An. cruzii  and  An. homunculus , which is a secondary malaria vector, are morphologically similar and difficult to distinguish when using external morphological characteristics only. These two species may occur syntopically with  An .  bellator , which is also a potential vector of  Plasmodium  species and is morphologically similar to  An. cruzii  and  An. homunculus . Identification of these species based on female specimens is often jeopardised by polymorphisms, overlapping morphological characteristics and damage caused to specimens during collection. Wing geometric morphometrics has been used to distinguish several insect species; however, this economical and powerful tool has not been applied to  Kerteszia  species. Our objective was to assess wing geometry to distinguish  An. cruzii ,  An. homunculus  and  An. bellator .    Methods  Specimens were collected in an area in the Serra do Mar hotspot biodiversity corridor of the Atlantic Forest biome (Cananeia municipality, State of Sao Paulo, Brazil). The right wings of females of  An. cruzii  (n= 40),  An. homunculus  (n= 50) and  An. bellator  (n= 27) were photographed. For each individual, 18 wing landmarks were subjected to standard geometric morphometrics. Discriminant analysis of Procrustean coordinates was performed to quantify wing shape variation.    Results  Individuals clustered into three distinct groups according to species with a slight overlap between representatives of  An. cruzii  and  An. homunculus . The Mahalanobis distance between  An. cruzii  and  An. homunculus  was consistently lower (3.50) than that between  An. cruzii  and  An. bellator  (4.58) or  An. homunculus  and  An. bellator  (4.32). Pairwise cross-validated reclassification showed that geometric morphometrics is an effective analytical method to distinguish between  An .  bellator ,  An. cruzii  and  An. homunculus  with a reliability rate varying between 78-88%. Shape analysis revealed that the wings of  An. homunculus  are narrower than those of  An. cruzii  and that  An. bellator  is different from both of the congeneric species.    Conclusion  It is possible to distinguish among the vectors  An. cruzii ,  An. homunculus  and  An. bellator  based on female wing characteristics.

Morphometrical diagnosis of the malaria vectors Anopheles cruzii, An. homunculus and An. bellator