The new Internal Transcribed Spacer 2 diagnostic tool clarifies the taxonomic position and geographic distribution of the North American malaria vector Anopheles punctipennis

The Eurasian malaria vector Anopheles messeae is a widely spread and genetically diverse species. Five widespread polymorphic chromosomal inversions were found in natural populations of this mosquito. A cryptic species, Anopheles daciae, was differentiated from An. messeae by the presence of several nucleotide substitutions in the Internal Transcribed Spacer 2 (ITS2) region of ribosomal DNA. However, because of the absence of a high-quality reference cytogenetic map, the inversion polymorphisms in An. daciae and An. messeae remain poorly understood. Moreover, a recently determined heterogeneity in ITS2 in An. daciae questioned the accuracy of the previously used Restriction Fragment Length Polymorphism (RFLP) assay for species diagnostics. In this study, a standard-universal cytogenetic map was constructed based on orcein stained images of chromosomes from salivary glands for population studies of the chromosomal inversions that can be used for both An. messeae and An. daciae. In addition, a new ITS2-RFLP approach for species diagnostics was developed. Both methods were applied to characterize inversion polymorphism in populations of An. messeae and An. daciae from a single location in Western Siberia in Russia. The analysis demonstrates that cryptic species are remarkably different in their frequencies of chromosomal inversion variants. Our study supports previous observations that An. messeae has higher inversion polymorphism in all autosomes than the cryptic species An. daciae.

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“…quadrimaculatus and An. crucians D, which can be present in the same location [ 65 ]. Similarly, the cryptic species An.…”

Section: Discussionmentioning

confidence: 99%

New Cytogenetic Photomap and Molecular Diagnostics for the Cryptic Species of the Malaria Mosquitoes Anopheles messeae and Anopheles daciae from Eurasia

Artemov

Fedorova

Karagodin

et al. 2021

Insects

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“…belenrae 26 , 27 . The ITS2-based PCR method has been used for distinguishing species in the Maculipennis Group 28 , e.g., the An. crucians complex 29 and others 30 .…”

Section: Introductionmentioning

confidence: 99%

Analysis of geometric morphometrics and molecular phylogeny for Anopheles species in the Republic of Korea

Jeon,

Kim,

Klein

et al. 2023

Sci Rep

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Human malaria, transmitted by Anopheles mosquitoes, is the most predominant mosquito-borne disease that is responsible for hundreds of thousands of deaths worldwide each year. In the Republic of Korea (ROK), there are currently several hundred malaria cases annually, mostly near the demilitarized zone (DMZ). Eight species of Anopheles mosquitoes are currently known to be present in the ROK. Similar to other major malaria vectors in Africa and India, it is very challenging to morphologically differentiate Anopheles mosquitoes in the ROK due to their extremely similar morphology. In this study, wing geometric morphometrics (WGM) were used to differentiate the eight Anopheles species collected at six locations near the DMZ, Seoul and Pyeongtaek from April–October 2021. Phylogenetic analysis was also performed using cytochrome c oxidase subunit 1 (COI), internal transcribed spacer 2 (ITS2), and tyrosine hydroxylase (TH) genes for comparison with WGM analysis and to infer evolutionary relationships. The results of cross-validation (overall accuracy = 74.8%) demonstrated that species identification using WGM alone was not possible with a high accuracy for all eight species. While phylogenetic analyses based on the COI region could not clearly distinguish some species, the analysis based on ITS2 and TH was more useful for resolving the phylogenetic correlation of the eight species. Our results may improve Anopheles species identification strategies for effective identification and control of malaria vectors in the ROK.

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Molecular characterization of anopheline species diversity in the Andaman and Nicobar archipelago, with a particular emphasis on Anopheles barbirostris

Pachalil,

Gupta,

Maile

et al. 2024

Parasitol Res

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The new Internal Transcribed Spacer 2 diagnostic tool clarifies the taxonomic position and geographic distribution of the North American malaria vector Anopheles punctipennis

Cited by 4 publications

References 33 publications

New Cytogenetic Photomap and Molecular Diagnostics for the Cryptic Species of the Malaria Mosquitoes Anopheles messeae and Anopheles daciae from Eurasia

New Cytogenetic Photomap and Molecular Diagnostics for the Cryptic Species of the Malaria Mosquitoes Anopheles messeae and Anopheles daciae from Eurasia

Analysis of geometric morphometrics and molecular phylogeny for Anopheles species in the Republic of Korea

Molecular characterization of anopheline species diversity in the Andaman and Nicobar archipelago, with a particular emphasis on Anopheles barbirostris

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