UNRAVELING THE PHENOTYPIC AND GENOTYPIC RESISTANCES ASSOCIATED WITH PYRETHROID AND ORGANOPHOSPHATE IN Aedes aegypti (DIPTERA: CULICIDAE)

Wan Fatma Zuharah, Ma Teng, Asfa Nurizzah Zin Azman, Fatin Nabila Abdullah

Abstract


The widespread emergence of insecticide resistance in Aedes aegypti threatens the effectiveness of dengue vector control programmes in Malaysia. This study investigated the phenotypic susceptibility and the underlying genotypic resistance mechanisms associated with pyrethroid and organophosphate insecticides in Ae. aegypti populations collected from four localities in Penang, Malaysia, namely Apartment Asoka (AA), Taman Bukit Jambul (TBJ), Taman Machang Bubok (TMB), and Flat Sri Pauh (FSP). Adult susceptibility bioassays were conducted following World Health Organization (WHO) protocols using deltamethrin (0.03%), permethrin (0.4%), pirimiphos-methyl (60 mg/m²), and malathion (5%). Target-site resistance was assessed by sequencing the voltage-gated sodium channel (VGSC) domains II and III. Bioassay results revealed severe and widespread pyrethroid resistance across all populations. Deltamethrin mortality ranged from 5% in Taman Bukit Jambul (TBJ) to 65% in Flat Sri Pauh (FSP), while permethrin mortality ranged from 3% (TBJ) to 85% (FSP), indicating operational failure of both insecticides. Pirimiphos-methyl showed moderate resistance, with mortality ranging from 30% to 65%, whereas malathion remained largely effective, achieving 100% mortality in most populations, except TBJ, which showed possible resistance (95%). Genotypic analysis identified widespread knockdown resistance (kdr) mutations, including S989P, V1016G, T1520I, and F1534C. The most resistant population (TBJ) exhibited complex mutation profiles, including triple-mutation genotypes (V1016G/T1520I/F1534C) at frequencies up to 20% under permethrin exposure. Notably, the T1520I mutation was detected predominantly in deltamethrin-exposed populations, with frequencies ranging from 14.3% to 25%, and rarely occurred as a single substitution. These findings indicate advanced resistance development in Ae. aegypti populations in Penang, underscoring the urgent need to revise pyrethroid-dependent control strategies and implement evidence-based insecticide resistance management.


Full Text:

PDF

References


Bhatt, S., Gething, P.W., Brady, O.J., Messina, J.P., Farlow, A.W., Moyes, C.L., et al. 2013. The global distribution and burden of dengue. Nature 496(7446): 504–507.

Brito, L.P., Carrara, L., Freitas, R.M.D., Lima, J.B.P. & Martins, A.J. 2018. Levels of resistance to pyrethroid among distinct kdr alleles in Aedes aegypti laboratory lines and frequency of kdr alleles in 27 natural populations from Rio de Janeiro, Brazil. BioMed research international 2018(1): 2410819.

Chen, M., Du, Y., Wu, S., Nomura, Y., Zhu, G., Zhorov, B.S. & Dong, K. 2019. Molecular evidence of sequential evolution of DDT- and pyrethroid-resistant sodium channels in Aedes aegypti. PLoS Neglected Tropical Diseases 13(6): e0007432.

Coleman, M. & Hemingway, J. 2007. Insecticide resistance monitoring and evaluation in disease-transmitting mosquitoes. Journal of Pesticide Science 32(2): 69–76.

Collins, E., Quintana, J., Morales, R., Martínez, K., Barrera, R. & Mackay, A.J. 2025. Profiling insecticide resistance phenotypes and genotypes in Aedes aegypti populations across four regions in Puerto Rico. Scientific Reports 15: 3709.

Du, Y., Nomura, Y., Zhorov, B.S. & Dong, K. 2016. Sodium channel mutations and pyrethroid resistance in Aedes aegypti. Insects 7(4): 60.

Fernando, H.S.D., Saavedra-Rodriguez, K., Perera, R. & Black, W.C. IV. 2020. Resistance to commonly used insecticides and underlying mechanisms of resistance in Aedes aegypti (L.) from Sri Lanka. Parasites & Vectors 13: 393.

Hien, A.S., Soma, D.D., Hema, O., Bayili, B., Namountougou, M., Gnankiné, O., Baldet, T., Diabaté, A. & Dabiré, K.R. 2017. Evidence that agricultural use of pesticides selects pyrethroid resistance within Anopheles gambiae sl populations from cotton growing areas in Burkina Faso, West Africa. PLoS One 12(3): e0173098.

Kawada, H., Higa, Y., Komagata, O., Kasai, S., Tomita, T., Nguyen, T.Y., et al. 2009. Widespread distribution of a newly found point mutation in the voltage-gated sodium channel in pyrethroid-resistant Aedes aegypti populations in Vietnam. PLoS Neglected Tropical Diseases 3(10): e527.

Kawada, H., Oo, S.Z.M., Thaung, S., Kawashima, E., Maung, Y.N.M., Thu, H.M., et al. 2016a. Co-occurrence of point mutations in the voltage-gated sodium channel of pyrethroid-resistant Aedes aegypti populations in Myanmar. PLoS Neglected Tropical Diseases 10(7): e0004765.

Kawada, H., Higa, Y., Futami, K., Muranami, Y., Kawashima, E., Osei, J.H.N., et al. 2016b. Discovery of point mutations in the voltage-gated sodium channel gene of Aedes aegypti populations in Myanmar. PLoS Neglected Tropical Diseases 10(7): e0004780.

Kushwah, R.B.S., Cherry L. D., Neera K., Tridibes A., & Singh O.P. 2015. Pyrethroid-resistance and presence of two knockdown resistance (kdr) mutations, F1534C and a novel mutation T1520I, in Indian Aedes aegypti. PLoS Neglected Tropical Diseases 9 (1):e3332.

Linss, J.G.B., Brito, L.P., Garcia, G.A., Araki, A.S., Bruno, R.V., Lima, J.B.P., et al. 2014. Distribution and dissemination of the Val1016Ile and Phe1534Cys knockdown resistance mutations in Brazilian Aedes aegypti populations. Parasites & Vectors 7: 25.

Liu, H., Xu, Q., Zhang, L. & Liu, N. 2005. Chlorpyrifos resistance in mosquito Culex quinquefasciatus. Journal of Medical Entomology 42(5): 815–820.

Moyes, C.L., Vontas, J., Martins, A.J., Ng, L.C., Koou, S.Y., Dusfour, I., et al. 2017. Contemporary status of insecticide resistance in the major Aedes vectors of arboviruses infecting humans. PLoS Neglected Tropical Diseases 11(7): e0005625.

Namountougou, M., Soma, D.D., Balboné, M., Kaboré, D.A., Kientega, M., Hien, A.S., et al. 2020. Monitoring insecticide susceptibility in Aedes aegypti populations from Burkina Faso: Implication of metabolic resistance. Tropical Medicine and Infectious Disease 5(2): 84.

Nurul-Nastasea, S., Yu, K-X., Raza, A., Mohamed Nor, Z., Tengku-Idris, T.I.N., Dianita, R., et al. 2023. Insecticide resistance status of Aedes aegypti and Aedes albopictus in Malaysia (2010–2022): A review. Asian Pacific Journal of Tropical Medicine 16(10): 434–445.

Raghavendra, K., Verma, V., Srivastava, H., Gunasekaran, K., Sreehari, U., & Dash, A.P. 2010. Persistence of DDT, malathion and deltamethrin resistance in Anopheles culicifacies after their withdrawal from indoor residual spraying in India. Indian Journal of Medical Research 132(3): 260–264.

Rubio-Palis, Y., Dzuris, N., Sandi, C., Vizcaino-Cabarrus, R.L., Corredor-Medina, C., González, J.A. & Lenhart, A.E. 2023. Insecticide resistance levels and associated mechanisms in three Aedes aegypti populations from Venezuela. Mem Inst Oswaldo Cruz 118: e220210.

Scott, J.G. 1990. Investigating mechanisms of insecticide resistance: Methods, strategies, and pitfalls. In Roush, R.T. & Tabashnik, B.E. (eds.). Pesticide Resistance in Arthropods, pp. 39–57). New York: Springer,

Silva, J.J., Kouam, C.N. & Scott, J.G. 2021. Levels of cross-resistance to pyrethroids conferred by the Vssc knockdown resistance allele 410L+1016I+1534C in Aedes aegypti. PLoS Neglected Tropical Diseases 15(7): e0009549.

Stenhouse, S.A., Plernsub, S., Yanola, J., Lumjuan, N., Dantrakool, A., Choochote, W. & Somboon, P. 2013. Detection of the V1016G mutation in the voltage-gated sodium channel gene of Aedes aegypti and its effect on deltamethrin resistance in Thailand. Parasites & Vectors 6: 253.

Tognarelli, J., Moya, P.R., González, C.R. & Collao-Ferrada, X. 2025. Global distribution and impact of knockdown resistance mutations in Aedes aegypti on pyrethroid resistance. Parasites & Vectors 18(1): 382.

Uemura, N., Itokawa, K., Komagata, O. & Kasai, S. 2024. Recent advances in the study of knockdown resistance (kdr) mutations in Aedes mosquitoes. Current Opinion in Insect Science 63: 101178.

Ullah, M.A. 2024. Breaking the fever: Global strategies for sustainable dengue recovery and prevention. Borneo Medical Journal 10(1): 1–3.

Wang, Y., Wang, X., Brown, D.J., An, M., Xue, R-D. & Liu, N. 2023. Insecticide resistance: Status and potential mechanisms in Aedes aegypti. Pesticide Biochemistry and Physiology 195: 105577.

World Health Organization (WHO). 2022. Manual For Monitoring Insecticide Resistance in Mosquito Vectors and Selecting Appropriate Interventions. Geneva: World Health Organization.

Yanola, J., Somboon, P., Walton, C., Nachaiwieng, W., Somwang, P. & Prapanthadara, L.A. 2011. High‐throughput assays for detection of the F1534C mutation in the voltage‐gated sodium channel gene in permethrin‐resistant Aedes aegypti and the distribution of this mutation throughout Thailand. Tropical Medicine & International Health 16(4): 501-509.

Zoh, M.G., Bonneville, J.M., Laporte, F., Tutagata, J., Sadia, C.G., Fodjo, B.K., Mouhamadou, C.S., McBeath, J., Schmitt, F., Horstmann, S. & Reynaud, S. 2023. Deltamethrin and transfluthrin select for distinct transcriptomic responses in the malaria vector Anopheles gambiae. Malaria Journal 22(1): 256.

Zongo, S., Toe, H.K., Guelbeogo, M.W., Sanou, A., Traore, A. & Dabiré, R.K. 2025. Spatial distribution of insecticide resistance in Aedes aegypti across Burkina Faso highlights strengthened resistance management strategies. Acta Tropica 271: 107847.

Zuharah, W.F. & Sufian, M. 2021. The discovery of a novel knockdown resistance (kdr) mutation A1007G on Aedes aegypti (Diptera: Culicidae) from Malaysia. Scientific Reports 11(1): 5180.


Refbacks

  • There are currently no refbacks.