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. 2014 Jan 15;9(1):e85496.
doi: 10.1371/journal.pone.0085496. eCollection 2014.

DNA barcoding for the identification of sand fly species (Diptera, Psychodidae, Phlebotominae) in Colombia

María Angélica Contreras Gutiérrez  1 , Rafael J Vivero  1 , Iván D Vélez  1 , Charles H Porter  2 , Sandra Uribe  3
Affiliations

DNA barcoding for the identification of sand fly species (Diptera, Psychodidae, Phlebotominae) in Colombia

María Angélica Contreras Gutiérrez et al. PLoS One. .

Abstract

Sand flies include a group of insects that are of medical importance and that vary in geographic distribution, ecology, and pathogen transmission. Approximately 163 species of sand flies have been reported in Colombia. Surveillance of the presence of sand fly species and the actualization of species distribution are important for predicting risks for and monitoring the expansion of diseases which sand flies can transmit. Currently, the identification of phlebotomine sand flies is based on morphological characters. However, morphological identification requires considerable skills and taxonomic expertise. In addition, significant morphological similarity between some species, especially among females, may cause difficulties during the identification process. DNA-based approaches have become increasingly useful and promising tools for estimating sand fly diversity and for ensuring the rapid and accurate identification of species. A partial sequence of the mitochondrial cytochrome oxidase gene subunit I (COI) is currently being used to differentiate species in different animal taxa, including insects, and it is referred as a barcoding sequence. The present study explored the utility of the DNA barcode approach for the identification of phlebotomine sand flies in Colombia. We sequenced 700 bp of the COI gene from 36 species collected from different geographic localities. The COI barcode sequence divergence within a single species was <2% in most cases, whereas this divergence ranged from 9% to 26.6% among different species. These results indicated that the barcoding gene correctly discriminated among the previously morphologically identified species with an efficacy of nearly 100%. Analyses of the generated sequences indicated that the observed species groupings were consistent with the morphological identifications. In conclusion, the barcoding gene was useful for species discrimination in sand flies from Colombia.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Map of the study area showing the sampling sites of sand flies collected in the present study.
Site 1: Jardín; Site 2: Jerico; Site 3: San Francisco; Site 4; San Roque; Site 5: Santa Fé de Antioquia; Site 6: Chinchina; Site 7: Salamina; Site 8: Samaná; Site 9: Victoria, Site 10: Acandí; Site 11: Bahía Solano, Playa Potes; Site 12: San Andrés de Sotavento; Site 13: Ricaute; Site 14: Quimbaya; Site 15: Gramalote; Site 16: Colosó, Serranía de Coraza Forestry Reserve; Site 17: Chaparral; Site 18: Alcalá; and Site 19: Caño Ariba.
Figure 2
Figure 2. Genetic divergences (K2P distances) between COI sequences for varied taxonomic levels of Colombian sand fly species.
The frequency of pairwise divergences among specimens both within species and among different species.
Figure 3
Figure 3. Neighbor-joining analysis of mitochondrial COI sequences of Colombian sand fly species.
See this image and copyright information in PMC

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