Detection of pyrethroid resistance mutations and intron variants in the voltage‐gated sodium channel of <i>Aedes (Stegomyia) aegypti</i> and <i>Aedes (Stegomyia) albopictus</i> mosquitoes from Lao People's Democratic Republic

Marcombe, Sébastien; Shimell, Katherine; Savage, Rachel; Howlett, Edward; Luangamath, Phonesavanh; Nilaxay, Somphat; Vungkyly, Vacky; Baby, Anne; King, Matthew R.; Clarke, Josie; Jeffries, Chloe; Jojo, Josna; Lacey, Emily; Bhatty, Farris; Mabika, Dadirayi; Cruz, Andrea Dela; Fisher, Cerys; Mbadu, Milca; Despiniadis, Iasonas; Brey, Paul T.; Thammavong, Phoutmany; Jones, Andrew K.

doi:10.1111/mve.12580

Cited by 8 publications

(20 citation statements)

References 52 publications

(116 reference statements)

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“…It is a sort of mystery that the L982W substitution has never been identified in the recent studies of A. aegypti from at least 16 regions of the neighboring countries: Thailand, Laos, and China, although this substitution was focused ( Fig. 4 ) ( 15 , 39 , 40 ). L982W was also not detected from 442 A. aegypti collected from three regions of Nepal in 2017 and 2018 ( 41 ).…”

Section: Discussionmentioning

confidence: 99%

“…S3 for the actual frequencies of all five alleles of Vgsc in the populations from Phnom Penh. No L982W substitution was detected from the cities marked with blue plots in China, Laos, Thailand, and Malaysia although the L982 was targeted (15,39,40). Note that L982W has never been detected in the Indochina Peninsula except Vietnam and Cambodia.…”

Section: Molecular Modeling Ofa Aegypti Vgscmentioning

confidence: 99%

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Discovery of super–insecticide-resistant dengue mosquitoes in Asia: Threats of concomitant knockdown resistance mutations

et al. 2022

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Aedes aegypti (Linnaeus, 1762) is the main mosquito vector for dengue and other arboviral infectious diseases. Control of this important vector highly relies on the use of insecticides, especially pyrethroids. The high frequency (>78%) of the L982W substitution was detected at the target site of the pyrethroid insecticide, the voltage-gated sodium channel (Vgsc) of A. aegypti collected from Vietnam and Cambodia. Alleles having concomitant mutations L982W + F1534C and V1016G + F1534C were also confirmed in both countries, and their frequency was high (>90%) in Phnom Penh, Cambodia. Strains having these alleles exhibited substantially higher levels of pyrethroid resistance than any other field population ever reported. The L982W substitution has never been detected in any country of the Indochina Peninsula except Vietnam and Cambodia, but it may be spreading to other areas of Asia, which can cause an unprecedentedly serious threat to the control of dengue fever as well as other Aedes -borne infectious diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Molecular Modeling Ofa Aegypti Vgscmentioning

confidence: 99%

Discovery of super–insecticide-resistant dengue mosquitoes in Asia: Threats of concomitant knockdown resistance mutations

et al. 2022

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“…The tenth intron 20 sequence consists of 237 bp and is identical to the previously characterized intron B variant [46] found in Ae. aegypti from Laos [37]. None of the intron variants observed here were identical to any of the four variants seen in Ae.…”

Section: Detection Of Mutations and Intron Variants In The Voltage-ga...mentioning

confidence: 38%

“…Mutation at the 410, 989, 1016, 1520, and 1534 amino acid positions in domains I, II, and III of the VGSC were detected in Asia in Ae. albopictus mosquitoes from China, Laos, Taiwan, Thailand, and Vietnam [30,[36][37][38].…”

Section: Introductionmentioning

confidence: 99%

Metabolic Resistance and Not Voltage-Gated Sodium Channel Gene Mutation Is Associated with Pyrethroid Resistance of Aedes albopictus (Skuse, 1894) from Cambodia

Marcombe,

Doeurk,

Thammavong

et al. 2024

Insects

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(1) Background: In Cambodia, Aedes albopictus is an important vector of the dengue virus. Vector control using insecticides is a major strategy implemented in managing mosquito-borne diseases. Resistance, however, threatens to undermine the use of insecticides. In this study, we present the levels of insecticide resistance of Ae. albopictus in Cambodia and the mechanisms involved. (2) Methods: Two Ae. albopictus populations were collected from the capital, Phnom Penh city, and from rural Pailin province. Adults were tested with diagnostic doses of malathion (0.8%), deltamethrin (0.03%), permethrin (0.25%), and DDT (4%) using WHO tube assays. Synergist assays using piperonyl butoxide (PBO) were implemented before the pyrethroid assays to detect the potential involvement of metabolic resistance mechanisms. Adult female mosquitoes collected from Phnom Penh and Pailin were tested for voltage-gated sodium channel (VGSC) kdr (knockdown resistance) mutations commonly found in Aedes sp.-resistant populations throughout Asia (S989P, V1016G, and F1534C), as well as for other mutations (V410L, L982W, A1007G, I1011M, T1520I, and D1763Y). (3) Results: The two populations showed resistance against all the insecticides tested (<90% mortality). The use of PBO (an inhibitor of P450s) strongly restored the efficacy of deltamethrin and permethrin against the two resistant populations. Sequences of regions of the vgsc gene showed a lack of kdr mutations known to be associated with pyrethroid resistance. However, four novel non-synonymous mutations (L412P/S, C983S, Q1554STOP, and R1718L) and twenty-nine synonymous mutations were detected. It remains to be determined whether these mutations contribute to pyrethroid resistance. (4) Conclusions: Pyrethroid resistance is occurring in two Ae. albopictus populations originating from urban and rural areas of Cambodia. The resistance is likely due to metabolic resistance specifically involving P450s monooxygenases. The levels of resistance against different insecticide classes are a cause for concern in Cambodia. Alternative tools and insecticides for controlling dengue vectors should be used to minimize disease prevalence in the country.

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“…In addition, five different introns were found in Domain III, of which two introns consisted of 68 and 70 bp, respectively, and the other three introns each consisted of 83 bp [ 52 ]. In the present study, introns A, B, and E were consistent with those in this previous study [ 52 ], although two other introns, referred to as C and D, were also found. The types of introns appear to play an important role in the regulation of gene mutation, but how this effect occurs remains unclear.…”

Section: Discussionmentioning

confidence: 99%

High frequency of Voltage-gated sodium channel (VGSC) gene mutations in Aedes albopictus (Diptera: Culicidae) suggest rapid insecticide resistance evolution in Shanghai, China

Yuan¹,

Shan²,

Zhang³

et al. 2023

PLoS Negl Trop Dis

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Background Dengue fever is an infectious disease that is imported into Shanghai, China and requires prevention and control measures. Controlling the vector Aedes albopictus through insecticide use is a key approach to dengue control. However, the rapid evolution of insecticide resistance in Ae. albopictus has raised concerns about the failure of dengue control efforts. Knockdown resistance (kdr) caused by point mutations in the voltage-gated sodium channel (VGSC) gene is a primary mechanism of pyrethroid resistance. In this study, we investigated the kdr mutations of Ae. albopictus in Shanghai and evaluated the trend in its evolution. Methodology/principal findings We collected 17 populations of Ae. albopictus from 15 districts in Shanghai in 2020, extracted genomic DNA from individual mosquitoes, and amplified Domain II, III, and IV in VGSC using PCR. Following sequencing, we obtained 658 VGSC sequences. We detected the nonsynonymous mutations V1016G, I1532T, and F1534S/C/I, among which V1016G and F1534C/I were reported in Shanghai for the first time and F1534I was a novel mutant allele in Ae. albopictus. The overall mutation frequency was 84.65%, with individual mutation frequencies ranging from 46.81% to 100%, excluding the Fengxian District population, which had a frequency of 0%. The V1016G and I1532T mutation types accounted for 7.14% and 3.42%, respectively. The mutant allele at codon 1534 accounted for 63.98% of all mutations, including TCC/S (62.77%), TGC/C (1.06%), and ATC/I (0.15%). We identified and classified five intron types in Domain III by length, including A (83 bp, 12.07%), B (68 bp, 87.30%), C (80 bp, 0.16%), D (72 bp, 0.16%), and E (70 bp, 0.31%). Individuals with intron B had a significant mutation tendency at codon 1534 relative to intron A (chi-square test, p < 0.0001). We found no correlation between mutation frequency and the amount of pyrethroid used (Pearson correlation, p = 0.4755). Conclusions/significance In recent years, kdr mutations in the Ae. albopictus population in Shanghai have rapidly evolved, as evidenced by an increase in mutation types and significantly increased mutation frequency. The F1534I/ATC mutant allele was found to be a novel mutation, F1534C/TGC was reported for the first time in Shanghai, and intron B in Domain III was significantly associated with mutation frequency at codon 1534. Continuous monitoring of resistance changes and strict regulation of insecticide use are required.

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Detection of pyrethroid resistance mutations and intron variants in the voltage‐gated sodium channel of Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus mosquitoes from Lao People's Democratic Republic

Cited by 8 publications

References 52 publications

Discovery of super–insecticide-resistant dengue mosquitoes in Asia: Threats of concomitant knockdown resistance mutations

Discovery of super–insecticide-resistant dengue mosquitoes in Asia: Threats of concomitant knockdown resistance mutations

Metabolic Resistance and Not Voltage-Gated Sodium Channel Gene Mutation Is Associated with Pyrethroid Resistance of Aedes albopictus (Skuse, 1894) from Cambodia

High frequency of Voltage-gated sodium channel (VGSC) gene mutations in Aedes albopictus (Diptera: Culicidae) suggest rapid insecticide resistance evolution in Shanghai, China

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