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DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka

BACKGROUND: Vectors of mosquito-borne diseases in Sri Lanka, except for malaria, belong to the subfamily Culicinae, which includes nearly 84% of the mosquito fauna of the country. Hence, accurate and precise species identification of culicine mosquitoes is a crucial factor in implementing effective...

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Autores principales: Weeraratne, Thilini Chathurika, Surendran, Sinnathamby Noble, Parakrama Karunaratne, S. H. P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5918568/
https://www.ncbi.nlm.nih.gov/pubmed/29695263
http://dx.doi.org/10.1186/s13071-018-2810-z
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author Weeraratne, Thilini Chathurika
Surendran, Sinnathamby Noble
Parakrama Karunaratne, S. H. P.
author_facet Weeraratne, Thilini Chathurika
Surendran, Sinnathamby Noble
Parakrama Karunaratne, S. H. P.
author_sort Weeraratne, Thilini Chathurika
collection PubMed
description BACKGROUND: Vectors of mosquito-borne diseases in Sri Lanka, except for malaria, belong to the subfamily Culicinae, which includes nearly 84% of the mosquito fauna of the country. Hence, accurate and precise species identification of culicine mosquitoes is a crucial factor in implementing effective vector control strategies. During the present study, a combined effort using morphology and DNA barcoding was made to characterize mosquitoes of the subfamily Culicinae for the first time from nine districts of Sri Lanka. Cytochrome c oxidase subunit 1 (cox1) gene from the mitochondrial genome and the internal transcribed spacer 2 (ITS2) region from the nuclear ribosomal DNA were used for molecular characterization. RESULTS: According to morphological identification, the field collected adult mosquitoes belonged to 5 genera and 14 species, i.e. Aedes aegypti, Ae. albopictus, Ae. pallidostriatus, Aedes sp. 1, Armigeres sp. 1, Culex bitaeniorhynchus, Cx. fuscocephala, Cx. gelidus, Cx. pseudovishnui, Cx. quinquefasciatus, Cx. tritaeniorhynchus, Cx. whitmorei, Mansonia uniformis and Mimomyia chamberlaini. Molecular analyses of 62 cox1 and 36 ITS2 sequences were exclusively comparable with the morphological identifications of all the species except for Ae. pallidostriatus and Aedes sp. 1. Although the species identification of Armigeres sp. 1 specimens using morphological features was not possible during this study, DNA barcodes of the specimens matched 100% with the publicly available Ar. subalbatus sequences, giving their species status. Analysis of all the cox1 sequences (14 clades supported by strong bootstrap value in the Neighbor-Joining tree and interspecific distances of > 3%) showed the presence of 14 different species. This is the first available DNA sequence in the GenBank records for morphologically identified Ae. pallidostriatus. Aedes sp. 1 could not be identified morphologically or by publicly available sequences. Aedes aegypti, Ae. albopictus and all Culex species reported during the current study are vectors of human diseases. All these vector species showed comparatively high diversity. CONCLUSIONS: The current study reflects the significance of integrated systematic approach and use of cox1 and ITS genetic markers in mosquito taxonomy. Results of DNA barcoding were comparable with morphological identifications and, more importantly, DNA barcoding could accurately identify the species in the instances where the traditional morphological identification failed due to indistinguishable characters of damaged specimens and the presence of subspecies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13071-018-2810-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-59185682018-04-30 DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka Weeraratne, Thilini Chathurika Surendran, Sinnathamby Noble Parakrama Karunaratne, S. H. P. Parasit Vectors Research BACKGROUND: Vectors of mosquito-borne diseases in Sri Lanka, except for malaria, belong to the subfamily Culicinae, which includes nearly 84% of the mosquito fauna of the country. Hence, accurate and precise species identification of culicine mosquitoes is a crucial factor in implementing effective vector control strategies. During the present study, a combined effort using morphology and DNA barcoding was made to characterize mosquitoes of the subfamily Culicinae for the first time from nine districts of Sri Lanka. Cytochrome c oxidase subunit 1 (cox1) gene from the mitochondrial genome and the internal transcribed spacer 2 (ITS2) region from the nuclear ribosomal DNA were used for molecular characterization. RESULTS: According to morphological identification, the field collected adult mosquitoes belonged to 5 genera and 14 species, i.e. Aedes aegypti, Ae. albopictus, Ae. pallidostriatus, Aedes sp. 1, Armigeres sp. 1, Culex bitaeniorhynchus, Cx. fuscocephala, Cx. gelidus, Cx. pseudovishnui, Cx. quinquefasciatus, Cx. tritaeniorhynchus, Cx. whitmorei, Mansonia uniformis and Mimomyia chamberlaini. Molecular analyses of 62 cox1 and 36 ITS2 sequences were exclusively comparable with the morphological identifications of all the species except for Ae. pallidostriatus and Aedes sp. 1. Although the species identification of Armigeres sp. 1 specimens using morphological features was not possible during this study, DNA barcodes of the specimens matched 100% with the publicly available Ar. subalbatus sequences, giving their species status. Analysis of all the cox1 sequences (14 clades supported by strong bootstrap value in the Neighbor-Joining tree and interspecific distances of > 3%) showed the presence of 14 different species. This is the first available DNA sequence in the GenBank records for morphologically identified Ae. pallidostriatus. Aedes sp. 1 could not be identified morphologically or by publicly available sequences. Aedes aegypti, Ae. albopictus and all Culex species reported during the current study are vectors of human diseases. All these vector species showed comparatively high diversity. CONCLUSIONS: The current study reflects the significance of integrated systematic approach and use of cox1 and ITS genetic markers in mosquito taxonomy. Results of DNA barcoding were comparable with morphological identifications and, more importantly, DNA barcoding could accurately identify the species in the instances where the traditional morphological identification failed due to indistinguishable characters of damaged specimens and the presence of subspecies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13071-018-2810-z) contains supplementary material, which is available to authorized users. BioMed Central 2018-04-25 /pmc/articles/PMC5918568/ /pubmed/29695263 http://dx.doi.org/10.1186/s13071-018-2810-z Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Weeraratne, Thilini Chathurika
Surendran, Sinnathamby Noble
Parakrama Karunaratne, S. H. P.
DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka
title DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka
title_full DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka
title_fullStr DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka
title_full_unstemmed DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka
title_short DNA barcoding of morphologically characterized mosquitoes belonging to the subfamily Culicinae from Sri Lanka
title_sort dna barcoding of morphologically characterized mosquitoes belonging to the subfamily culicinae from sri lanka
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5918568/
https://www.ncbi.nlm.nih.gov/pubmed/29695263
http://dx.doi.org/10.1186/s13071-018-2810-z
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