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

Freddy Ruiz-Lopez1*, Richard C Wilkerson1, Jan E Conn23, Sascha N McKeon3, David M Levin1, Martha L Quiñones4, Marinete M Póvoa5 and Yvonne-Marie Linton16

  • * Corresponding author: Freddy Ruiz-Lopez ruizj@si.edu

  • † Equal contributors

Author Affiliations

1 Entomology Branch, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, USA

2 Griffin Laboratory, Wadsworth Center, New York State Department of Health, Albany, New York, USA

3 Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, New York, USA

4 Facultad de Medicina, Universidad Nacional de Colombia, Bogotá D.C., Colombia

5 Instituto Evandro Chagas, Ananindeua, Pará, Brazil

6 Department of Entomology, Natural History Museum, London, UK

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Parasites & Vectors 2012, 5:44  doi:10.1186/1756-3305-5-44

Published: 21 February 2012

Abstract

Background

Mosquitoes 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.

Methods

DNA 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.

Results

Species 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.

Conclusions

DNA 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.

Keywords:
Albitarsis Group; barcoding; COI; new species; An. albitarsis G; An. albitarsis H; An. albitarsis I