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Deep sequencing of pyrethroid-resistant bed bugs reveals multiple mechanisms of resistance within a single population

ABSTRACT: A frightening resurgence of bed bug infestations has occurred over the last 10 years in the US. Current chemical methods have been inadequate for controlling bed bugs due to widespread insecticide resistance. Little is known about the mechanisms of resistance present in US bed bug populations, making it extremely difficult to develop intelligent strategies to control this pest. We have identified bed bugs collected in Richmond, VA which exhibit both kdr-type (L925I) and metabolic resistance to pyrethroid insecticides. LD50 bioassays determined resistance ratios of ~6000-fold to the insecticide deltamethrin, with contact bioassays confirming cross-resistance to several other labeled formulations. To identify metabolic genes potentially involved in the detoxification of pyrethroids, we performed deep-sequencing of the adult bed bug transcriptome, obtaining more than 2.5 million reads on the 454 titanium platform. Following assembly, analysis of newly identified gene transcripts in both Harlan (susceptible) and Richmond (resistant) bed bugs revealed several candidate cytochrome P450 and carboxyesterase genes which were significantly over-expressed in the resistant strain, consistent with the idea of increased metabolic resistance. These data will accelerate efforts to understand the biochemical basis for insecticide resistance in bed bugs, and provide molecular markers to assist in the surveillance of metabolic resistance.

ORGANISM(S): Cimex lectularius

PROVIDER: GSE31025 | GEO | 2011/10/19

SECONDARY ACCESSION(S): PRJNA144889

REPOSITORIES: GEO

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Deep sequencing of pyrethroid-resistant bed bugs reveals multiple mechanisms of resistance within a single population

Project description:A frightening resurgence of bed bug infestations has occurred over the last 10 years in the US. Current chemical methods have been inadequate for controlling bed bugs due to widespread insecticide resistance. Little is known about the mechanisms of resistance present in US bed bug populations, making it extremely difficult to develop intelligent strategies to control this pest. We have identified bed bugs collected in Richmond, VA which exhibit both kdr-type (L925I) and metabolic resistance to pyrethroid insecticides. LD50 bioassays determined resistance ratios of ~6000-fold to the insecticide deltamethrin, with contact bioassays confirming cross-resistance to several other labeled formulations. To identify metabolic genes potentially involved in the detoxification of pyrethroids, we performed deep-sequencing of the adult bed bug transcriptome, obtaining more than 2.5 million reads on the 454 titanium platform. Following assembly, analysis of newly identified gene transcripts in both Harlan (susceptible) and Richmond (resistant) bed bugs revealed several candidate cytochrome P450 and carboxyesterase genes which were significantly over-expressed in the resistant strain, consistent with the idea of increased metabolic resistance. These data will accelerate efforts to understand the biochemical basis for insecticide resistance in bed bugs, and provide molecular markers to assist in the surveillance of metabolic resistance. Deep sequencing was performed from total RNA isolated from adult male bed bugs using the Titanium 454 platform

2011-10-19 | E-GEOD-31025 | biostudies-arrayexpress

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Transcriptome analysis of insecticide resistant housefly strain: insights about SNPs and regulatory elements in differentially expressed cytochrome P450 genes

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Transcriptome analysis of insecticide resistant housefly strain: insights about SNPs and regulatory elements in differentially expressed cytochrome P450 genes

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Nkya et al 2014 - Insecticide resistance mechanisms associated with different environments in the malaria vector Anopheles gambiae. A case study in Tanzania

Project description:Nine Anopheles gambiae populations were sampled in three areas of Tanzania showing contrasting agriculture activity, urbanization and usage of insecticides for vector control. Insecticide resistance levels were measured in larvae and adults through bioassays with deltamethrin, DDT and bendiocarb. A microarray approach was used for identifying transcription level variations associated to different environments and insecticide resistance. the Ifakara strain originating from central Tanzania and susceptible to all insecticides was used as a reference strain.

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Anopheles funestus amidst a South/North shift in the role of key resistance genes across Malawi

Project description:Resistance to pyrethroids, the only insecticide approved for bednets, threatens control of the major malaria vector, Anopheles funestus, in Malawi. To improve the management of such resistance countrywide, it is crucial to understand the dynamics and mechanisms driving resistance in the field. In this study the levels of insecticide resistance were determined across the highly endemic densely populated lake and southern agricultural area. Insecticide resistance to pyrethoids was assessed using standardized WHO bioassay methods and resistant mosquitoes were hybridized to susceptible mosquitoes. This microarray analysis revealed the key role of cytochrome P450 genes such as CYP6P9a, CYP6P9b and CYP6M7. However, a significant shift in the over-expression of these CYP450s was detected across a south/north transect, with CYP6M7 more highly over-transcribed in the two northern collection sites and the tandemly duplicated genes, CYP6P9a and CYP6P9b, more greatly over-transcribed in the south.

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Molecular bases of P450-mediated resistance to the neonicotinoid insecticide imidacloprid in the mosquito Ae. aegypti

Project description:Resistance to chemical insecticides including pyrethroids, the main insecticide class used against mosquitoes, led to a regain of interest for neonicotinoids. In this context, the present study aims at characterizing the molecular basis of neonicotinoid resistance in the mosquito Aedes aegypti. Resistance mechanisms were studied by combining transcriptomic and genomic data obtained from a laboratory strain selected at the larval stage for 30 generations with imidacloprid (Imida-R line). After thirty generations of selection, larvae of Imida-R line showed a 8-fold increased resistance to imidacloprid and a significant cross-tolerance to the pyrethroids permethrin and deltamethrin. Cross-resistance to pyrethroids was only observed in adults when larvae were previously exposed to imidacloprid suggesting a low but inducible expression of resistance alleles at the adult stage. Resistance of the Imida-R line was associated with a slower larval development time in females. Multiple detoxification enzymes were over-transcribed in larvae in association with resistance including the P450s CYP6BB2, CYP9M9 and CYP6M11 previously associated with pyrethroid resistance. Some of them together with their redox partner NADPH P450 reductase were also affected by non-synonymous mutations associated with resistance. Combining genomic and transcriptomic data allowed identifying promoter variations associated with the up-regulation of CYP6BB2 in the resistant line. Overall, these data confirm the key role of P450s in neonicotinoid resistance in Ae. aegypti and their potential to confer cross-resistance to pyrethroids, raising concerns about the use of neonicotinoids for resistance management in this mosquito species.

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Impact of environmental pollutants on insecticide-driven gene selection in mosquitoes

Project description:This microarray study aimed at evaluating the impact of mosquito chemical environment on the selection of insecticide resistance mechanisms. Here the mosquito Aedes aegypti was used as a model to perform a laboratory experiment combining mosquito larvae exposure to a sub-lethal dose of xenobiotic and their selection with the insecticide permethrin. After ten generations, bioassays and a transcriptome profiling with the 15K microarray Aedes detox chip plus microarray were performed comparatively on all strains.

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Proteomics studies confirm CYP450 core involvement in multiple resistance strains of Aedes aegypti from French Guiana

Project description:Insecticide resistance is a worldwide threat for vector control around the world, and Aedes aegypti , the main vector of several arboviruses, is a particular concern. To better understand the mechanisms of resistance, four isofemale strains originally from French Guiana were isolated and analysed using combined approaches. The activity of detoxification enzymes involved in insecticide resistance was assayed, and mutations located at positions 1016 and 1534 of the sodium voltage-gated channel gene, which have been associated with pyrethroid resistance in Aedes aegypti populations in Latin America, were monitored. Resistance to other insecticide families (organophosphates and carbamates) was evaluated. A large-scale proteomic analysis was performed to identify proteins involved in insecticide resistance. Our results revealed a metabolic resistance and resistance associated with a mutation of the sodium voltage-gated channel gene at position 1016. Metabolic resistance was mediated through an increase of esterase activity in most strains but also through the shifts in the abundance of several cytochrome P450 (CYP450s). Overall, resistance to deltamethrin was linked in the isofemale strains to resistance to other class of insecticides, suggesting that cross- and multiple resistance occur through selection of mechanisms of metabolic resistance. These results give some insights into resistance to deltamethrin and into multiple resistance phenomena in populations of Ae. aegypti

2020-12-09 | PXD020408 | Pride

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