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Phylogeography of the neotropical Anopheles triannulatus complex (Diptera: Culicidae) supports deep structure and complex patterns

Marta Moreno113*, Sara Bickersmith1, Wesley Harlow1, Jessica Hildebrandt1, Sascha N McKeon12, Teresa Fernandes Silva-do-Nascimento3, Jose R Loaiza4, Freddy Ruiz5, Ricardo Lourenço-de-Oliveira3, Maria AM Sallum6, Eduardo S Bergo7, Gary N Fritz8, Richard C Wilkerson5, Yvonne M Linton5, Maria J Dantur Juri9, Yadira Rangel10, Marinete M Póvoa11, Lina A Gutiérrez-Builes12, Margarita M Correa12 and Jan E Conn12

Author Affiliations

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

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

3 Departamento de Entomología, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro, Brazil

4 Instituto de Investigaciones Científicas y Servicios de Alta Tecnología, Clayton, Panamá, República de Panamá

5 Division of Entomology, Walter Reed Army Institute of Research, Silver Spring, MD, USA

6 Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, São Paulo, Brazil

7 Superintendência de Controle de Endemias, SUCEN, São Paulo, Brazil

8 Department of Biological Sciences, Eastern Illinois University, Charleston, IL, USA

9 Instituto Superior de Entomología "Dr. Abraham Willink", Facultad de Ciencias, Naturales e Instituto Miguel Lillo, Universidad Nacional de Tucumán, Tucumán, Argentina

10 Laboratorio de Biologia de Vectores, Instituto de Zoología y Ecología Tropical, Universidad Central de Venezuela, Caracas, Venezuela

11 Instituto Evandro Chagas, Secção de psitologia, Belém, Brazil

12 Grupo de Microbiología Molecular, Escuela de Microbiología, Universidad de Antioquia, Medellín, Colombia

13 Present address: Division Infectious Diseases University of California San Diego, George Palade Labs, School of Medicine, 92093, 9500 Gilman Drive, MC 0741, La Jolla, CA, USA

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Parasites & Vectors 2013, 6:47  doi:10.1186/1756-3305-6-47

Published: 22 February 2013



The molecular phylogenetic relationships and population structure of the species of the Anopheles triannulatus complex: Anopheles triannulatus s.s., Anopheles halophylus and the putative species Anopheles triannulatus C were investigated.


The mitochondrial COI gene, the nuclear white gene and rDNA ITS2 of samples that include the known geographic distribution of these taxa were analyzed. Phylogenetic analyses were performed using Bayesian inference, Maximum parsimony and Maximum likelihood approaches.


Each data set analyzed septely yielded a different topology but none provided evidence for the seption of An. halophylus and An. triannulatus C, consistent with the hypothesis that the two are undergoing incipient speciation. The phylogenetic analyses of the white gene found three main clades, whereas the statistical parsimony network detected only a single metapopulation of Anopheles triannulatus s.l. Seven COI lineages were detected by phylogenetic and network analysis. In contrast, the network, but not the phylogenetic analyses, strongly supported three ITS2 groups. Combined data analyses provided the best resolution of the trees, with two major clades, Amazonian (clade I) and trans-Andean + Amazon Delta (clade II). Clade I consists of multiple subclades: An. halophylus + An. triannulatus C; trans-Andean Venezuela; central Amazonia + central Bolivia; Atlantic coastal lowland; and Amazon delta. Clade II includes three subclades: Panama; cis-Andean Colombia; and cis-Venezuela. The Amazon delta specimens are in both clades, likely indicating local sympatry. Spatial and molecular variance analyses detected nine groups, corroborating some of subclades obtained in the combined data analysis.


Combination of the three molecular markers provided the best resolution for differentiation within An. triannulatus s.s. and An. halophylus and C. The latest two species seem to be very closely related and the analyses performed were not conclusive regarding species differentiation. Further studies including new molecular markers would be desirable to solve this species status question. Besides, results of the study indicate a trans-Andean origin for An. triannulatus s.l. The potential implications for malaria epidemiology remain to be investigated.

Anopheles triannulatus s.s.; Anopheles halophylus; Anopheles triannulatus C; phylogeography; COI gene; White gene; ITS2