Taxonomia integrativa, filogeografia e análise de infectividade por Plasmodium ssp. no Complexo Oswaldoi-Konderi (DIPTERA: CULICIDAE: ANOPHELINAE)

Abstract

Anopheles oswaldoi s.l. (Peryassú) was long considered a single species, which would be implicated as a secondary vector of malaria in some localities of the Amazon region. Subsequent study described An. konderi Galvão & Damasceno, counting only on differences in the apex of the aedeagus of male genitália. Recent analyzes using molecular approaches revealed that these two species are in fact a complex of cryptic species, with wide geographical distribution and extensive sympatric ranges between them. Currently believed that five species (An. oswaldoi s.s., An. konderi s.s., An. oswaldoi A, An. oswaldoi B and An. sp. nr. konderi) exist in this named complex of Oswaldoi-Konderi. However, these species had never been analyzed morphologically, and despite being implicated as secondary vectors, the molecular identity of these species found with Plasmodium had not yet been determined. In the present study, 29 localities of the Brazilian Amazon were sampled, 71 spawns were obtained and studied morphologically, to explore the diversity of species in the complex and reveal reliable morphological characters for its determination. For this, mitochondrial inheritance genetic markers were used, such as the Cytochrome c Oxidase Subunit I (COI) gene and a nuclear inheritance marker, Second Transcribed Internal Spacer (ITS2). The numbers of operating taxonomic units (OTU's) were inferred, the phylogenetic relationships recovered using the Bayesian Inference and the time of diversification of the Oswaldoi-Konderi complex in the Amazon basin was determined. Our results indicated a high number of operating taxonomic units (14 - 18 OTU's). The morphology supported five species based on male genitalia characters. The Neotype and Paratypes of An. konderi s.s. which were reexamined, indicated a mixture of two morphological forms, confirmed with the molecular data, including another nuclear marker, the CAD gene, which is linked to the X chromosome but presents low heterozygosity. Anopheles tadei n. sp. was described based on morphological differences between male genitals and two characters in the larva near An. konderi. The analysis of a larger region of the COI gene (1.211bp) in An. oswaldoi A indicated structuring for three populations, Para, Amazonas, and Roraima, respectively. Further studies will be needed to elucidate the taxonomic identity of An. oswaldoi A and An. oswaldoi B, as An. aquacaelestis Curry has not been examined and is synonymous with An. oswaldoi s.s.