Molecular characterization and identification of members of the Anopheles subpictus complex in Sri Lanka

Surendran, Sinnathamby N.; Sarma, Devojit Kumar; Jude, Pavilupillai J.; Kemppainen, Petri; Kanthakumaran, Nadarajah; Gajapathy, K.; Peiris, Lalanthika B S; Ramasamy, Ranjan; Walton, Catherine

doi:10.1186/1475-2875-12-304

Cited by 48 publications

(62 citation statements)

References 34 publications

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“…sundaicus than An. subpictus species A, as observed in previous studies [18]. This indicates the considerable genetic distance between these two species and supports the above statement.…”

Section: Discussionsupporting

confidence: 91%

Molecular characterization of Anopheline (Diptera: Culicidae) mosquitoes from eight geographical locations of Sri Lanka

Weeraratne

Surendran

Reimer

et al. 2017

Malar J

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BackgroundGenus Anopheles is a major mosquito group of interest in Sri Lanka as it includes vectors of malaria and its members exist as species complexes. Taxonomy of the group is mainly based on morphological features, which are not conclusive and can be easily erased while handling the specimens. A combined effort, using morphology and DNA barcoding (using the markers cytochrome c oxidase subunit I (COI) gene and internal transcribed spacer 2 (ITS2) region, was made during the present study to recognize anophelines collected from eight districts of Sri Lanka for the first time.Methods Cytochrome c oxidase subunit I and ITS2 regions of morphologically identified anopheline mosquitoes from Sri Lanka were sequenced. These sequences together with GenBank sequences were used in phylogenetic tree construction and molecular characterization of mosquitoes.ResultsAccording to morphological identification, the field-collected adult mosquitoes belonged to 15 species, i.e., Anopheles aconitus, Anopheles annularis, Anopheles barbirostris, Anopheles culicifacies, Anopheles jamesii, Anopheles karwari, Anopheles maculatus, Anopheles nigerrimus, Anopheles pallidus, Anopheles peditaeniatus, Anopheles pseudojamesi, Anopheles subpictus, Anopheles tessellatus, Anopheles vagus, and Anopheles varuna. However, analysis of 123 COI sequences (445 bp) (16 clades supported by strong bootstrap value in the neighbour joining tree and inter-specific distances of >3%) showed that there are 16 distinct species. Identity of the morphologically identified species, except An. subpictus, was comparable with the DNA barcoding results. COI sequence analysis showed that morphologically identified An. subpictus is composed of two genetic entities: An. subpictus species A and species B (inter-specific K2P distance 0.128). All the four haplotypes of An. culicifacies discovered during the present study belonged to a single species. ITS2 sequences (542 bp) were obtained for all the species except for An. barbirostris, An. subpictus species B, An. tessellatus, and An. varuna. Each of these sequences was represented by a single species-specific haplotype.ConclusionsThe present study reflects the importance and feasibility of COI and ITS2 genetic markers in identifying anophelines and their sibling species, and the significance of integrated systematic approach in mosquito taxonomy. Wide distribution of malaria vectors in the country perhaps indicates the potential for re-emergence of malaria in the country.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-017-1876-y) contains supplementary material, which is available to authorized users.

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“…sundaicus than An. subpictus species A, as observed in previous studies [18]. This indicates the considerable genetic distance between these two species and supports the above statement.…”

Section: Discussionsupporting

confidence: 91%

Molecular characterization of Anopheline (Diptera: Culicidae) mosquitoes from eight geographical locations of Sri Lanka

Weeraratne

Surendran

Reimer

et al. 2017

Malar J

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“…All the mosquitoes sequenced for ITS2 rDNA (n ¼ 24) had identical sequence and were 100% similar to "Species A" of Sri Lanka (Surendran et al 2013). In the absence of chromosomal inversion genotype, we consider them as putative species A. Sequencing of 28S-d3 rDNA (n ¼ 10) also did not reveal inter-individual difference and had 100% homology with sequence data of Sri Lankan species A (Singh et al, unpublished), which is suggestive of presence of only one species.…”

Section: Resultsmentioning

confidence: 99%

“…has been recognized as a complex of four sibling species, A through D, identified on the basis of arrangement of two inversions present on polytene chromosome X (Suguna 1982, Suguna et al 1994 and also on the morphological basis (Reuben andSuguna 1983, Suguna et al 1994). However, doubt has been raised on the validity of such morphological characteristics (Subbarao et al 1988;Surendran et al 2013). The sequencing of ITS2-rDNA and 28S rDNA of randomly selected samples of An.…”

Section: Discussionmentioning

confidence: 99%

L1014F-kdr Mutation in Indian Anopheles subpictus (Diptera: Culicidae) Arising From Two Alternative Transversions in the Voltage-Gated Sodium Channel and a Single PIRA-PCR for Their Detection

Singh

Dykes

Sharma

et al. 2015

Journal of Medical Entomology

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Leucine-to-phenylalanine substitution at residue L1014 in the voltage-gated sodium channel, target site of action for dichlorodiphenyltrichloroethane (DDT) and pyrethroids, is the most common knockdown resistance (kdr) mutation reported in several insects conferring resistance against DDT and pyrethroids. Here, we report presence of two coexisting alternative transversions, A>T and A>C, on the third codon position of L1014 residue in malaria vector Anopheles subpictus Grassi (species A) from Jamshedpur (India), both leading to the same amino acid substitution of Leu-to-Phe with allelic frequencies of 19 and 67%, respectively. A single primer-introduced restriction analysis-polymerase chain reaction (PIRA-PCR) was devised for the identification of L1014F-kdr mutation in An. subpictus resulting from either type of point mutation. Genotyping of samples with PIRA-PCR revealed high frequency (82%) of L1014F-kdr mutation in the study area.

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“…DNA was extracted from larvae or legs of the adult mosquito using the modified phenol-chloroform method in Surendran et al (2013). For mosquitoes from the Americas, DNA was extracted from whole adults using the method of Livak (1984).…”

Section: Methodsmentioning

confidence: 99%

Historical environmental change in Africa drives divergence and admixture of Aedes aegypti mosquitoes: a precursor to successful worldwide colonization?

et al. 2016

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Increasing globalization has promoted the spread of exotic species, including disease vectors. Understanding the evolutionary processes involved in such colonizations is both of intrinsic biological interest and important to predict and mitigate future disease risks. The Aedes aegypti mosquito is a major vector of dengue, chikungunya and Zika, the worldwide spread of which has been facilitated by Ae. aegypti's adaption to human-modified environments. Understanding the evolutionary processes involved in this invasion requires characterization of the genetic make-up of the source population (s). The application of approximate Bayesian computation (ABC) to sequence data from four nuclear and one mitochondrial marker revealed that African populations of Ae. aegypti best fit a demographic model of lineage diversification, historical admixture and recent population structuring. As ancestral Ae. aegypti were dependent on forests, this population history is consistent with the effects of forest fragmentation and expansion driven by Pleistocene climatic change. Alternatively, or additionally, historical human movement across the continent may have facilitated their recent spread and mixing. ABC analysis and haplotype networks support earlier inferences of a single out-of-Africa colonization event, while a cline of decreasing genetic diversity indicates that Ae. aegypti moved first from Africa to the Americas and then to Asia. ABC analysis was unable to verify this colonization route, possibly because the genetic signal of admixture obscures the true colonization pathway. By increasing genetic diversity and forming novel allelic combinations, divergence and historical admixture within Africa could have provided the adaptive potential needed for the successful worldwide spread of Ae. aegypti.

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Molecular characterization and identification of members of the Anopheles subpictus complex in Sri Lanka

Cited by 48 publications

References 34 publications

Molecular characterization of Anopheline (Diptera: Culicidae) mosquitoes from eight geographical locations of Sri Lanka

Molecular characterization of Anopheline (Diptera: Culicidae) mosquitoes from eight geographical locations of Sri Lanka

L1014F-kdr Mutation in Indian Anopheles subpictus (Diptera: Culicidae) Arising From Two Alternative Transversions in the Voltage-Gated Sodium Channel and a Single PIRA-PCR for Their Detection

Historical environmental change in Africa drives divergence and admixture of Aedes aegypti mosquitoes: a precursor to successful worldwide colonization?

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