Pyrethroid resistance inAedes albopictusin Pudong New Area, Shanghai China

Abstract

Objective: To evaluate the phenotypic resistance profile of Aedes (Ae.) albopictus against multiple insecticides and to delineate the frequency and pyrethroid-resistance association of knockdown resistance (kdr) mutations V1016G, I1532T and F1534S in Ae. albopictus in Pudong New Area, Shanghai, China.

Methods: From June to July 2024, a total of 1 249 Ae. albopictus eggs and 4 732 adults were collected from five towns in the Pudong New Area. These collected specimens were then reared to obtain the F1 generation including 10 235 larvae at the late 3rd to early 4th instar stages, as well as 5 800 non-blood-fed adults aged 3 to 5 days. Larval resistance was assessed via the immersion method, and adult resistance via the contact tube method. For the larval, 0≤resistance ratio (RR)<3 indicates sensitivity; 3≤RR<10 denotes low resistance; 10≤RR<40 signifies moderate resistance; RR≥40 represents high resistance. While for adults, a mortality rate of ≥98% stands for sensitivity, 80%≤mortality rate<98% indicates suspected resistance, and mortality rate<80% denotes a resistant population. A pooled sequencing strategy (8 sample pools) was used to detect genotype kdr mutations including V1016G, I1532T, F1534S in the voltage-gated sodium channel (VGSC) gene.

Results: Ae. albopictus larvae displayed low resistance to fenthion (RR=5.85), temephos (RR=4.63) and β-cypermethrin (RR=3.20), but remained fully susceptible to propoxur (RR=0.33). Adults showed incipient resistance to β-cypermethrin (mortality: 92.7%), lambda-cyhalothrin (mortality: 96.8%) and permethrin (mortality: 86.6%), while remained sensitive to propoxur (mortality: 100%). For Ae. albopictus population in the Pudong New Area, pyrethroid tolerance was primarily driven by F1534S mutation, which was amplified when the mutation was paired with V1016G or I1532T.

Conclusions: These findings highlight the need for integrated resistance monitoring and targeted insecticide rotation—such as prioritizing propoxur for Ae. albopictus larval and adult control—to mitigate resistance spread and preserve vector management efficacy.

Keywords

Pyrethroid / Resistance / Knockdown resistance gene / Resistance phenotype

References

[1] Borgherini G, Poubeau P, Jossaume A, Gouix A, Cotte L, Michault A, et al. Persistent arthralgia associated with chikungunya virus: A study of 88 adult patients on Réunion Island. Clin Infect Dis 2008; 47(4): 469-475.

[2] de Souza WM, Ribeiro GS, de Lima STS, de Jesus R, Moreira FRR, Whittaker C, et al. Chikungunya: A decade of burden in the Americas. Lancet Reg Health Am 2024; 30: 100673.

[3] Suhrbier A. Rheumatic manifestations of chikungunya: Emerging concepts and interventions. Nat Rev Rheumatol 2019; 15: 597-611.

[4] Pan J, Fang C, Yan J, Yan H, Zhan B, Sun Y, et al. Chikungunya fever outbreak, Zhejiang province, China, 2017. Emerg Infect Dis 2019; 25(8): 1589-1591.

[5] Yin X, Hu T, Zhang H, Liu Y, Zhou Z, Liu L, et al. Emergent chikungunya fever and vertical transmission in Yunnan Province, China, 2019. Arch Virol 2021; 166(5): 1455-1462.

[6] Ramesh K, Divya M, Sakshi C, Debasis N. Sujatha sunil impact of CHIKV replication on the global proteome of Aedes albopictus cells. Proteomes 2022; 10(4): 38.

[7] Sahu MC, Samantaray RK, Pal A, Pati S. Recent advances on pathogenesis, diagnosis, prevention, immunological aspects, and vectors of dengue: A review. Asian Pac J Trop Biomed 2023; 13(8): 325-338.

[8] Zhou X, Li J, Ni R, Qiu X, Zhang Y, Tong Y. Insecticide resistance in the field populations of the Asian tiger mosquito Aedes albopictus in Beijing: Resistance status and associated detoxification genes. Front Physiol 2024; 15: 1498313.

[9] Liu N. Insecticide resistance in mosquitoes: Impact, mechanisms, and research directions. Annu Rev Entomol 2015; 60: 537-559.

[10] Elia-Amira NMR, Chen CD, Lau KW, Low VL, Lee HL, Azidah AA, et al. Adulticide resistance status of Aedes albopictus (Diptera: Culicidae) in Sabah, Malaysia: A statewide assessment. J Med Entomol 2022; 59(5): 1719-1726.

[11] Black WC, Snell TK, Saavedra-Rodriguez K, Kading RC, Campbell CL. From global to local-new insights into features of pyrethroid detoxification in vector mosquitoes. Insects 2021; 12(4): 276. doi: 10.3390/insects12040276.

[12] Thanispong K, Sathantriphop S, Chareonviriyaphap T. Establishment of diagnostic doses of five pyrethroids for monitoring physiological resistance in Aedes albopictus in Thailand. J Am Mosq Control Assoc 2015; 31(4): 346-350.

[13] Kasai S, Caputo B, Tsunoda T, Cuong TC, Maekawa Y, Lam-Phua SG, et al. First detection of a Vssc allele V1016G conferring a high level of insecticide resistance in Aedes albopictus collected from Europe (Italy) and Asia (Vietnam), 2016: A new emerging threat to controlling arboviral diseases. Euro Surveill 2019; 24(5): 1700847.

[14] Chinese Center for Disease Control and Prevention. Guidelines for vector mosquito control technology. Beijing: People’s Medical Publishing House; 2022.

[15] Zhou X, Yang C, Liu N, Li M, Tong Y, Zeng X, et al. Knockdown resistance (kdr) mutations within seventeen field populations of Aedes albopictus from Beijing, China: First report of a novel V1016G mutation and evolutionary origins of kdr haplotypes. Parasit Vectors 2019; 12(1): 180.

[16] Black WC 4th, Snell TK, Saavedra-Rodriguez K, Kading RC, Campbell CL. From global to local—new insights into features of pyrethroid detoxification in vector mosquitoes. Insects 2021; 12(4): 276.

[17] Smith LB, Kasai S, Scott JG. Pyrethroid resistance in Aedes aegypti and Aedes albopictus: Important mosquito vectors of human diseases. Pestic Biochem Physiol 2016; 133: 1-12. doi: 10.1016/j.pestbp.2016.03.005.

[18] Xu J, Bonizzoni M, Zhong D, Zhou G, Cai S, Li Y, et al. Multi-country survey revealed prevalent and novel F1534S mutation in voltage-gated sodium channel (VGSC) gene in Aedes albopictus. PLoS Negl Trop Dis 2016; 10(5): e0004696.

[19] Pichler V, Malandruccolo C, Serini P, Bellini R, Severini F, Toma L, et al. Phenotypic and genotypic pyrethroid resistance of Aedes albopictus, with focus on the 2017 chikungunya outbreak in Italy. Pest Manag Sci 2019; 75(10): 2642-2651.

[20] Kasai S, Caputo B, Tsunoda T, Cuong TC, Maekawa Y, Lam-Phua SG, et al. First detection of a Vssc allele V1016G conferring a high level of insecticide resistance in Aedes albopictus collected from Europe (Italy) and Asia (Vietnam), 2016: A new emerging threat to controlling arboviral diseases. Euro Surveill 2019; 24(5): 1700847.

[21] Gao JP, Chen HM, Shi H, Peng H, Ma YJ. Correlation between adult pyrethroid resistance and knockdown resistance (kdr) mutations in Aedes albopictus (Diptera: Culicidae) field populations in China. Infect Dis Poverty 2018; 7(1): 86.

[22] Zhou X, Yang C, Liu N, Li M, Tong Y, Zeng X, et al. Knockdown resistance (kdr) mutations within seventeen field populations of Aedes albopictus from Beijing China: First report of a novel V1016G mutation and evolutionary origins of kdr haplotypes. Parasit Vectors 2019; 12(1): 180.

[23] Chen H, Li K, Wang X, Yang X, Lin Y, Cai F, et al. First identification of kdr allele F1534S in VGSC gene and its association with resistance to pyrethroid insecticides in Aedes albopictus populations from Haikou City, Hainan Island, China. Infect Dis Poverty 2016; 5: 31.

[24] Plernsub S, Saingamsook J, Yanola J, Lumjuan N, Tippawangkosol P, Sukontason K, et al. Additive effect of knockdown resistance mutations, S989P, V1016G and F1534C, in a heterozygous genotype conferring pyrethroid resistance in Aedes aegypti in Thailand. Parasit Vectors 2016; 9(1): 417.

[25] Hirata K, Komagata O, Itokawa K, Yamamoto A, Tomita T, Kasai S. A single crossing-over event in voltage-sensitive Na⁺ channel genes may cause critical failure of chikungunya mosquito control by insecticides. PLoS Negl Trop Dis 2014; 8(8): e3085.

[26] Zhang S, Xu H, Xia S, Zhang L, Zhang L, Liu H. Analysis on the resistance of Aedes albopictus to insecticides in Putuo District, Shanghai. Shanghai J Prev Med 2022; 34(2): 113-118. (in Chinese).

[27] Darsie RF, Ward RA. Identification and geographical distribution of the mosquitoes of North America, North of Mexico. Gainesville: University Press of Florida; 2005, p. 45-52.

[28] Menegon M, Severini F, Toma L, Martignoni M, Di Luca M. Rapid molecular method for early detection of the invasive mosquito Aedes aegypti (Linnaeus, 1762) at points of entry. Acta Trop 2025; 264: 107605.

[29] Wei Y, Zheng X, He S, Xin X, Zhang J, Hu K. Insecticide susceptibility status and knockdown resistance (kdr) mutation in Aedes albopictus in China. Parasit Vectors 2021; 14(1): 609.

[30] Standardization Administration of the People's Republic of China, General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China. The bioassay methods for mosquitoes' resistance to insecticides: GB/T 29566-2013. Beijing: Standards Press of China; 2013.

[31] Rath A, Mohanty I, Hazra RK. Insecticide susceptibility status of invasive Aedes albopictus across chikungunya endemic districts of Odisha, India. Pest Manag Sci 2018; 74(6): 1431-1440.

[32] Wei Y, Zheng X, He S, Xin X, Zhang J, Hu K. Insecticide susceptibility status and knockdown resistance (kdr) mutation in Aedes albopictus in China. Parasit Vectors 2021; 14(1): 609.

[33] Mashlawi AM, Al-Nazawi AM, Noureldin EM, Alqahtani H, Mahyoub JA, Saingamsook J. Molecular analysis of knockdown resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes aegypti populations from Saudi Arabia. Parasit Vectors 2022; 15(1): 375.

[34] Wu S, Zhang Y, Li Y, Chen X, Liu Q, Xu J. Study on sensitivity and knockdown resistance genes of Aedes albopictus to pyrethroid insecticides in Xiamen, Fujian province, China, 2020. Chin J Vector Biol Control 2022; 33(2): 177-182. (in Chinese)

[35] Zhu C, Zhang Y, Li Y, Chen X, Liu Q, Xu J. Distribution of knockdown resistance genotypes in Aedes albopictus in Jinghong, Yunnan province, China, 2018-2019. Chin J Vector Biol Control 2020; 31(1): 7-11. (in Chinese)

[36] Li Y, Zhang Y, Chen X, Liu Q, Xu J, Wang H. Knockdown resistance gene mutations of Aedes albopictus from Fuzhou and Putian, Fujian, 2020. China Trop Med 2021; 21(10): 952-955, 969. (in Chinese)

[37] Liu H, Liu L, Cheng P, Yang L, Chen J, Lu Y. Bionomics and insecticide resistance of Aedes albopictus in Shandong, a high latitude and high-risk chikungunya transmission area in China. Parasit Vectors 2020; 13(1): 11.

[38] Zhou XJ, Zhang Y, Li Y, Chen X, Liu Q, Xu J. Detection and analysis of kdr resistance mutations in two populations of Aedes albopictus in Beijing. Chin J Hyg 2021; 27(4): 304-307. (in Chinese)

[39] Wu Y, Liu Q, Qi Y, Wu Y, Ni Q, Chen W. Knockdown resistance (kdr) mutations I1532T and F1534S were identified in Aedes albopictus field populations in Zhejiang Province, Central China. Front Cell Infect Microbiol 2021; 11: 702081.

[40] Li Y, Zhou G, Zhong D, Wang X, Hemming-Schroeder E, David RE. Widespread multiple insecticide resistance in the major chikungunya vector Aedes albopictus in Hainan Province, China. Pest Manag Sc 2021; 77(4): 1945-1953.