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EVOLUTION OF KDR HAPLOTYPES IN WORLDWIDE POPULATIONS OF AEDES AEGYPTI: INDEPENDENT ORIGINS OF THE F1534C KDR MUTATION
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Yale University. New Haven, CT, USA.
Yale University. New Haven, CT, USA / Center for Vector Biology & Zoonotic Diseases. The Connectivut Agricultural Experiment Station. New Haven, CT, USA.
Yale University. New Haven, CT, USA.
Yale University. New Haven, CT, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Fisiologia e Controle de Artrópodes Vetores. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.
Yale University. New Haven, CT, USA / Center for Vector Biology & Zoonotic Diseases. The Connectivut Agricultural Experiment Station. New Haven, CT, USA.
Yale University. New Haven, CT, USA.
Yale University. New Haven, CT, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Fisiologia e Controle de Artrópodes Vetores. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.
Abstract
Aedes aegypti is the primary vector of dengue, chikungunya, Zika, and urban yellow fever.
Insecticides are often the most effective tools to rapidly decrease the density of vector populations,
especially during arbovirus disease outbreaks. However, the intense use of insecticides,
particularly pyrethroids, has selected for resistant mosquito populations worldwide.
Mutations in the voltage gated sodium channel (NaV) are among the principal mechanisms
of resistance to pyrethroids and DDT, also known as “knockdown resistance,” kdr. Here we
report studies on the origin and dispersion of kdr haplotypes in samples of Ae. aegypti from
its worldwide distribution. We amplified the IIS6 and IIIS6 NaV segments from pools of Ae.
aegypti populations from 15 countries, in South and North America, Africa, Asia, Pacific,
and Australia. The amplicons were barcoded and sequenced using NGS Ion Torrent. Output
data were filtered and analyzed using the bioinformatic pipeline Seekdeep to determine frequencies
of the IIS6 and IIIS6 haplotypes per population. Phylogenetic relationships among
the haplotypes were used to infer whether the kdr mutations have a single or multiple origin.
We found 26 and 18 haplotypes, respectively for the IIS6 and IIIS6 segments, among which
were the known kdr mutations 989P, 1011M, 1016I and 1016G (IIS6), 1520I, and 1534C
(IIIS6). The highest diversity of haplotypes was found in African samples. Kdr mutations
1011M and 1016I were found only in American and African populations, 989P + 1016G and
1520I + 1534C in Asia, while 1534C was present in samples from all continents, except Australia.
Based primarily on the intron sequence, IIS6 haplotypes were subdivided into two
well-defined clades (A and B). Subsequent phasing of the IIS6 + IIIS6 haplotypes indicates
two distinct origins for the 1534C kdr mutation. These results provide evidence of kdr mutations
arising de novo at specific locations within the Ae. aegypti geographic distribution. In
addition, our results suggest that the 1534C kdr mutation had at least two independent origins.
We can thus conclude that insecticide selection pressure with DDT and more recently
with pyrethroids is selecting for independent convergent mutations in NaV.
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