Pyrethroid Resistance and Gene Expression Profile of a New Resistant Colony from Uganda Reveals Multiple Resistance Mechanisms and Overexpression of Glutathione-S-Transferases Linked to Survival of PBO-pyrethroid Combination

Overview

Journal Wellcome Open Res

Specialty Biomedical Engineering

Date 2024 May 30

PMID 38813466

Authors

Ambrose Oruni

Amy Lynd

Harun Njoroge

Ismail Onyige

Arjen E Vant Hof

Enock Matovu

Martin J Donnelly

Affiliations

Soon will be listed here.

Abstract

Background: The effectiveness of long-lasting insecticidal nets (LLINs) are being threatened by growing resistance to pyrethroids. To restore their efficacy, a synergist, piperonyl butoxide (PBO) which inhibits cytochrome P450s has been incorporated into pyrethroid treated nets. A trial of PBO-LLINs was conducted in Uganda from 2017 and we attempted to characterize mechanisms of resistance that could impact intervention efficacy.

Methods: We established an s.s colony in 2018 using female mosquitoes collected from Busia district in eastern Uganda. We first assessed the phenotypic resistance profile of this colony using WHO tube and net assays using a deltamethrin dose-response approach. The Busia colony was screened for known resistance markers and RT-qPCR targeting 15 genes previously associated with insecticide resistance was performed.

Results: The Busia colony had very high resistance to deltamethrin, permethrin and DDT. In addition, the colony had moderate resistance to alpha-cypermethrin and lambda-cyhalothrin but were fully susceptible to bendiocarb and fenitrothion. Exposure to PBO in combination with permethrin and deltamethrin resulted in higher mortality rates in both net and tube assays, with a higher mortality observed in net assays than tube assays. The marker, was at very high frequency (91.7-98.9%) whilst the metabolic markers and were at very low (1.3% - 11.5%) and moderate (39.5% - 44.7%) frequencies respectively. Our analysis showed that gene expression pattern in mosquitoes exposed to deltamethrin, permethrin or DDT only were similar in comparison to the susceptible strain and there was significant overexpression of cytochrome P450s, glutathione-s-transferases (GSTs) and carboxyl esterases (COEs). However, mosquitoes exposed to both PBO and pyrethroid strikingly and significantly only overexpressed closely related GSTs compared to unexposed mosquitoes while major cytochrome P450s were underexpressed.

Conclusions: The high levels of pyrethroid resistance observed in Busia appears associated with a wide range of metabolic gene families.

Citing Articles

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Oruni A, Arinaitwe E, Adiga J, Otto G, Kyagamba P, Okoth J bioRxiv. 2025; .

PMID: 40027642 PMC: 11870430. DOI: 10.1101/2025.02.13.638152.

Temporal evolution of insecticide resistance and bionomics in Anopheles funestus, a key malaria vector in Uganda.

Oruni A, Tchouakui M, Tagne C, Hearn J, Kayondo J, Wondji C Sci Rep. 2024; 14(1):32027.

PMID: 39738472 PMC: 11685729. DOI: 10.1038/s41598-024-83689-6.

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