DNA barcoding reveals both known and novel taxa in the Albitarsis Group (Anopheles: Nyssorhynchus) of Neotropical malaria vectors

Abstract

BackgroundMosquitoes belonging to the Albitarsis Group (Anopheles: Nyssorhynchus) are of importance as malaria vectors across the Neotropics. The Group currently comprises six known species, and recent studies have indicated further hidden biodiversity within the Group. DNA barcoding has been proposed as a highly useful tool for species recognition, although its discriminatory utility has not been verified in closely related taxa across a wide geographic distribution.MethodsDNA barcodes (658 bp of the mtDNA Cytochrome c Oxidase - COI) were generated for 565 An. albitarsis s.l. collected in Argentina, Brazil, Colombia, Paraguay, Trinidad and Venezuela over the past twenty years, including specimens from type series and type localities. Here we test the utility of currently advocated barcoding methodologies, including the Kimura-two-parameter distance model (K2P) and Neighbor-joining analysis (NJ), for determining species delineation within mosquitoes of the Neotropical Albitarsis Group of malaria vectors (Anopheles: Nyssorhynchus), and compare results with Bayesian analysis.ResultsSpecies delineation through barcoding analysis and Bayesian phylogenetic analysis, fully concur. Analysis of 565 sequences (302 unique haplotypes) resolved nine NJ tree clusters, with less than 2% intra-node variation. Mean intra-specific variation (K2P) was 0.009 (range 0.002 - 0.014), whereas mean inter-specific divergence were several-fold higher at 0.041 (0.020 - 0.056), supporting the reported "barcoding gap". These results show full support for separate species status of the six known species in the Albitarsis Group (An. albitarsis s.s., An. albitarsis F, An. deaneorum, An. janconnae, An. marajoara and An. oryzalimnetes), and also support species level status for two previously detected lineages - An. albitarsis G &An. albitarsis I (designated herein). In addition, we highlight the presence of a unique mitochondrial lineage close to An. deaneorum and An. marajoara (An. albitarsis H) from Rondônia and Mato Grosso in southwestern Brazil. Further integrated studies are required to confirm the status of this lineage.ConclusionsDNA barcoding provides a reliable means of identifying both known and undiscovered biodiversity within the closely related taxa of the Albitarsis Group. We advocate its usage in future studies to elucidate the vector competence and respective distributions of all eight species in the Albitarsis Group and the novel mitochondrial lineage (An. albitarsis H) recovered in this study.

Ecological Suitability and Spatial Distribution of Five Anopheles Species in Amazonian Brazil

“…oryzalimnetes abundance and water chemistry may indicate a local adaptation to more saline waters around the Amazon and on the eastern perimeter of Brazil, which corresponds to its proposed distribution. 37 The ability to tolerate salinity varies among species, and both salinity and conductivity have been correlated with the presence or development quality (size, rate of emergence) of Anopheles larvae in Africa 1 and Latin America.…”

“…janconnae, all specimens initially identified as members of the Albitarsis Complex from 2009 and a subset from 2011 (randomly selected to ensure less than a 2.5% error rate for species delimitation) were sequenced by the Applied Genomics Technology Core (Wadsworth Center) for the barcode region and compared with samples from Ruiz and collaborators. 37 Total genomic DNA was extracted using the DNAeasy tissue kit (Qiagen, Valencia, CA) and preserved at the Wadsworth Center at −80 C. Habitat characterization. To achieve greater diversity of larval habitats, sampling was done once for each breeding site.…”

“…goeldii) has been described as a species readily able to colonize and dominate in altered environments 33 as a result of its ability to survive in a variety of conditions, such as sunlit or shaded sites, fresh, clear, still, or flowing water 34 and occasionally in turbid and polluted sites. 35 The Albitarsis complex consists of at least eight species and a new lineage 36,37 ; only some of these have been incriminated as malaria vectors. The medical importance of Anopheles oryzalimnetes (Wilkerson and Motoki), previously Anopheles albitarsis B, is largely unknown, although this species has been attracted to human bait.…”

Evaluation of criteria for species delimitation of bagworm moths (Lepidoptera: Psychidae)

“…The K2P is the most commonly used and widely accepted substitution model for DNA barcoding (Park et al, 2011;Ruiz-Lopez et al, 2012;Jin et al, 2013). Nevertheless, its applicability is under debate.…”

Phylogenetic signal at the Cytb-SertRNA-IG1-ND1 mitochondrial region in Anopheles (Kerteszia) neivai Howard, Dyar & Knab, 1913

“…The use of mitochondrial DNA (mtDNA) has played a vital role in understanding the phylogenetic relationships in Anopheles, including diversification patterns, divergence time estimation (13,14) and the recognition of cryptic species complexes (15)(16)(17). There are several advantages to using mtDNA, including variable mutation rates in eukaryotes, lack of introns, and high abundance of copies, which makes mtDNA relatively easy to obtain even from degraded samples (18).…”