CYP450 core involvement in multiple resistance strains of Aedes aegypti from French Guiana highlighted by proteomics, molecular and biochemical studies

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.

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Insecticide resistance status of Aedes aegypti in Bangladesh

Parasites & Vectors ◽

10.1186/s13071-020-04503-6 ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Hasan Mohammad Al-Amin ◽

Fatema Tuj Johora ◽

Seth R. Irish ◽

Muhammad Riadul Haque Hossainey ◽

Lucrecia Vizcaino ◽

...

Keyword(s):

Aedes Aegypti ◽

Insecticide Resistance ◽

Pyrethroid Resistance ◽

Control Strategies ◽

Resistance Mechanisms ◽

Capital City ◽

Detoxification Enzymes ◽

Knockdown Resistance ◽

Metabolic Resistance ◽

Pyrethroid Insecticides

Abstract Background Arboviral diseases, including dengue and chikungunya, are major public health concerns in Bangladesh where there have been unprecedented levels of transmission reported in recent years. The primary approach to control these diseases is to control the vector Aedes aegypti using pyrethroid insecticides. Although chemical control has long been practiced, no comprehensive analysis of Ae. aegypti susceptibility to insecticides has been conducted to date. The aim of this study was to determine the insecticide resistance status of Ae. aegypti in Bangladesh and investigate the role of detoxification enzymes and altered target site sensitivity as resistance mechanisms. Methods Eggs of Aedes mosquitoes were collected using ovitraps from five districts across Bangladesh and in eight neighborhoods of the capital city Dhaka, from August to November 2017. CDC bottle bioassays were conducted for permethrin, deltamethrin, malathion, and bendiocarb using 3- to 5-day-old F0–F2 non-blood-fed female mosquitoes. Biochemical assays were conducted to detect metabolic resistance mechanisms, and real-time PCR was performed to determine the frequencies of the knockdown resistance (kdr) mutations Gly1016, Cys1534, and Leu410. Results High levels of resistance to permethrin were detected in all Ae. aegypti populations, with mortality ranging from 0 to 14.8% at the diagnostic dose. Substantial resistance continued to be detected against higher (2×) doses of permethrin (5.1–44.4% mortality). Susceptibility to deltamethrin and malathion varied between populations while complete susceptibility to bendiocarb was observed in all populations. Significantly higher levels of esterase and oxidase activity were detected in most of the test populations as compared to the susceptible reference Rockefeller strain. A significant association was detected between permethrin resistance and the presence of Gly1016 and Cys1534 homozygotes. The frequency of kdr (knockdown resistance) alleles varied across the Dhaka Aedes populations. Leu410 was not detected in any of the tested populations. Conclusions The detection of widespread pyrethroid resistance and multiple resistance mechanisms highlights the urgency for implementing alternate Ae. aegypti control strategies. In addition, implementing routine monitoring of insecticide resistance in Ae. aegypti in Bangladesh will lead to a greater understanding of susceptibility trends over space and time, thereby enabling the development of improved control strategies.

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Insecticide resistance status of Aedes aegypti in Bangladesh

10.1101/2020.07.31.231076 ◽

2020 ◽

Author(s):

Hasan Mohammad Al-Amin ◽

Fatema Tuj Johora ◽

Seth R. Irish ◽

Muhammad Riadul Haque Hossainey ◽

Lucrecia Vizcaino ◽

...

Keyword(s):

Aedes Aegypti ◽

Insecticide Resistance ◽

Pyrethroid Resistance ◽

Control Strategies ◽

Resistance Mechanisms ◽

Capital City ◽

Detoxification Enzymes ◽

Health Concern ◽

Metabolic Resistance ◽

Pyrethroid Insecticides

AbstractBackgroundArboviral diseases including dengue and chikungunya are major public health concern in Bangladesh, with unprecedented levels of transmission reported in recent years. The primary approach to control these diseases is control of Aedes aegypti using pyrethroid insecticides. Although chemical control is long-practiced, no comprehensive analysis of Ae. aegypti susceptibility to insecticides has previously been conducted. This study aimed to determine the insecticide resistance status of Ae. aegypti in Bangladesh and investigate the role of detoxification enzymes and altered target site sensitivity as resistance mechanisms.MethodsAedes eggs were collected using ovitraps from five districts across the country and in eight neighborhoods of the capital city Dhaka from August to November 2017. CDC bottle bioassays were conducted for permethrin, deltamethrin, malathion, and bendiocarb using 3-5-day old F0-F2 non-blood fed female mosquitoes. Biochemical assays were conducted to detect metabolic resistance mechanisms and real-time PCR was performed to determine the frequencies of the knockdown resistance (kdr) mutations Gly1016, Cys1534, and Leu410.ResultsHigh levels of resistance to permethrin were detected in all Ae. aegypti populations, with mortality ranging from 0 – 14.8% at the diagnostic dose. Substantial resistance continued to be detected against higher (2X) doses of permethrin (5.1 – 44.4% mortality). Susceptibility to deltamethrin and malathion varied between populations while complete susceptibility to bendiocarb was observed in all populations. Significantly higher levels of esterase and oxidase activity were detected in most of the test populations as compared to the susceptible reference Rockefeller strain. A significant association was detected between permethrin resistance and the presence of Gly1016 and Cys1534 homozygotes. The frequency of kdr alleles varied across the Dhaka populations, and Leu410 was not detected in any of the tested populations.ConclusionsThe detection of widespread pyrethroid resistance and multiple mechanisms highlights the urgency for implementing alternate Ae. aegypti control strategies. In addition, implementing routine monitoring of insecticide resistance in Ae. aegypti in Bangladesh will lead to a greater understanding of susceptibility trends over space and time, thereby enabling the development of improved control strategies.

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Transcriptomic analysis of insecticide resistance in the lymphatic filariasis vectorCulex quinquefasciatus

10.1101/589028 ◽

2019 ◽

Author(s):

Walter Fabricio Silva Martins ◽

Craig Stephen Wilding ◽

Alison Taylor Isaacs ◽

Emily Joy Rippon ◽

Karine Megy ◽

...

Keyword(s):

Insecticide Resistance ◽

Lymphatic Filariasis ◽

Fold Increase ◽

Target Site ◽

Detoxification Enzymes ◽

Site Mutation ◽

Public Health Importance ◽

Metabolic Resistance ◽

Vector Borne Diseases ◽

Bendiocarb Resistance

ABSTRACTCulex quinquefasciatusplays an important role in transmission of vector-borne diseases of public health importance, including lymphatic filariasis (LF), as well as many arboviral diseases. Currently, efforts to tackleC. quinquefasciatusvectored diseases are based on either mass drug administration (MDA) for LF, or insecticide-based interventions. Widespread and intensive insecticide usage has resulted in increased resistance in mosquito vectors, includingC. quinquefasciatus. Herein, the transcriptome profile of Ugandan bendiocarb-resistantC. quinquefasciatuswas explored to identify candidate genes associated with insecticide resistance. Resistance to bendiocarb in exposed mosquitoes was marked, with 2.04% mortality following 1h exposure and 58.02% after 4h. Genotyping of the G119SAce-1target site mutation detected a highly significant association (p<0.0001; OR=25) between resistance andAce1-119S. However, synergist assays using the P450 inhibitor PBO or the esterase inhibitor TPP resulted in markedly increased mortality (to ≈80%), suggesting a role of metabolic resistance in the resistance phenotype. Using a novel, custom 60K whole-transcriptome microarray 16 genes significantly overexpressed in resistant mosquitoes were detected, with the P450Cyp6z18showing the highest differential gene expression (>8-fold increase vs unexposed controls). These results provide evidence that bendiocarb-resistance in UgandanC. quinquefasciatusis mediated by both target-site mechanisms and over-expression of detoxification enzymes.

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Knockdown resistance (kdr) of the voltage-gated sodium channel gene of Aedes aegypti population in Denpasar, Bali, Indonesia

Parasites & Vectors ◽

10.1186/s13071-017-2215-4 ◽

2017 ◽

Vol 10 (1) ◽

Cited By ~ 26

Author(s):

Penny Humaidah Hamid ◽

Joko Prastowo ◽

Anis Widyasari ◽

Anja Taubert ◽

Carlos Hermosilla

Keyword(s):

Aedes Aegypti ◽

Sodium Channel ◽

Knockdown Resistance ◽

Voltage Gated Sodium Channel ◽

Voltage Gated ◽

Channel Gene ◽

Sodium Channel Gene ◽

Aegypti Population

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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 ◽

10.1111/j.1365-3156.2011.02725.x ◽

2011 ◽

Vol 16 (4) ◽

pp. 501-509 ◽

Cited By ~ 111

Author(s):

Jintana Yanola ◽

Pradya Somboon ◽

Catherine Walton ◽

Woottichai Nachaiwieng ◽

Puckavadee Somwang ◽

...

Keyword(s):

Aedes Aegypti ◽

Sodium Channel ◽

High Throughput ◽

Voltage Gated Sodium Channel ◽

Voltage Gated ◽

Channel Gene ◽

Sodium Channel Gene

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Detection of the V1016G mutation in the voltage-gated sodium channel gene of Aedes aegypti (Diptera: Culicidae) by allele-specific PCR assay, and its distribution and effect on deltamethrin resistance in Thailand

Parasites & Vectors ◽

10.1186/1756-3305-6-253 ◽

2013 ◽

Vol 6 (1) ◽

pp. 253 ◽

Cited By ~ 65

Author(s):

Steven A Stenhouse ◽

Suriya Plernsub ◽

Jintana Yanola ◽

Nongkran Lumjuan ◽

Anchalee Dantrakool ◽

...

Keyword(s):

Aedes Aegypti ◽

Sodium Channel ◽

Pcr Assay ◽

Voltage Gated ◽

Channel Gene ◽

Specific Pcr ◽

Deltamethrin Resistance ◽

Allele Specific ◽

Specific Pcr Assay ◽

Allele Specific Pcr

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A novel F1552/C1552 point mutation in the Aedes aegypti voltage-gated sodium channel gene associated with permethrin resistance

Pesticide Biochemistry and Physiology ◽

10.1016/j.pestbp.2009.10.005 ◽

2010 ◽

Vol 96 (3) ◽

pp. 127-131 ◽

Cited By ~ 55

Author(s):

Jintana Yanola ◽

Pradya Somboon ◽

Catherine Walton ◽

Woottichai Nachaiwieng ◽

La-aied Prapanthadara

Keyword(s):

Aedes Aegypti ◽

Sodium Channel ◽

Point Mutation ◽

Voltage Gated Sodium Channel ◽

Voltage Gated ◽

Channel Gene ◽

Sodium Channel Gene

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Insecticide Resistance Status of Aedes aegypti (Diptera: Culicidae) in California by Biochemical Assays

Journal of Medical Entomology ◽

10.1093/jme/tjaa031 ◽

2020 ◽

Vol 57 (4) ◽

pp. 1176-1183

Author(s):

Fan Yang ◽

Samuel Schildhauer ◽

Sarah A Billeter ◽

Melissa Hardstone Yoshimizu ◽

Robert Payne ◽

...

Keyword(s):

Aedes Aegypti ◽

Insecticide Resistance ◽

Southern California ◽

Mosquito Control ◽

Central Valley ◽

Activity Levels ◽

Metabolic Resistance ◽

Organophosphate Resistance ◽

Biochemical Assays ◽

Effective Use

Abstract Insecticide resistance in Aedes aegypti mosquitoes poses a major threat to public health worldwide. There are two primary biological mechanisms that can lead to insecticide resistance, target site and metabolic resistance, both of which confer resistance to specific classes of insecticides. Due to the limited number of chemical compounds available for mosquito control, it is important to determine current enzymatic profiles among mosquito populations. This study assessed resistance profiles for three metabolic pathways, α-esterases, β-esterases, and mixed-function oxidases (MFOs), as well as insensitivity of the acetylcholinesterase (iAChE) enzyme in the presence of propoxur, among Ae. aegypti from the Central Valley and southern California. All field-collected Ae. aegypti demonstrated elevated MFOs and iAChE activity, indicating potential development of pyrethroid and organophosphate resistance, respectively. Although regional variations were found among α-esterase and β-esterase activity, levels were generally elevated, further suggesting additional mechanisms for developing organophosphate resistance. Furthermore, mosquito samples from southern California exhibited a higher expression level to all three metabolic enzymes and iAChE activity in comparison to mosquitoes from the central region. These results could help guide future mosquito control efforts, directing the effective use of insecticides while limiting the spread of resistance.

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Monitoring Insecticide Susceptibility in Aedes Aegypti Populations from the Two Biggest Cities, Ouagadougou and Bobo-Dioulasso, in Burkina Faso: Implication of Metabolic Resistance

Tropical Medicine and Infectious Disease ◽

10.3390/tropicalmed5020084 ◽

2020 ◽

Vol 5 (2) ◽

pp. 84

Author(s):

Moussa Namountougou ◽

Dieudonné Diloma Soma ◽

Mahamoudou Balboné ◽

Didier Alexandre Kaboré ◽

Mahamadi Kientega ◽

...

Keyword(s):

Cytochrome P450 ◽

Aedes Aegypti ◽

West Africa ◽

Insecticide Resistance ◽

Burkina Faso ◽

Urban Areas ◽

Mortality Rates ◽

Metabolic Resistance ◽

Bobo Dioulasso ◽

Glutathione S Transferases

In West Africa, Aedes aegypti remains the major vector of dengue virus. Since 2013, dengue fever has been reemerging in Burkina Faso with annual outbreaks, thus becoming a major public health problem. Its control relies on vector control, which is unfortunately facing the problem of insecticide resistance. At the time of this study, although data on phenotypic resistance were available, information related to the metabolic resistance in Aedes populations from Burkina Faso remained very scarce. Here, we assessed the phenotypic and the metabolic resistance of Ae. aegypti populations sampled from the two main urban areas (Ouagadougou and Bobo-Dioulasso) of Burkina Faso. Insecticide susceptibility bioassays to chlorpyriphos-methyl 0.4%, bendiocarb 0.1% and deltamethrin 0.05% were performed on natural populations of Ae. aegypti using the WHO protocol. The activity of enzymes involved in the rapid detoxification of insecticides, especially non-specific esterases, oxidases (cytochrome P450) and glutathione-S-transferases, was measured on individual mosquitos. The mortality rates for deltamethrin 0.05% were low and ranged from 20.72% to 89.62% in the Bobo-Dioulasso and Ouagadougou sites, respectively. When bendiocarb 0.1% was tested, the mortality rates ranged from 7.73% to 71.23%. Interestingly, in the two urban areas, mosquitoes were found to be fully susceptible to chlorpyriphos-methyl 0.4%. Elevated activity of non-specific esterases and glutathione-S-transferases was reported, suggesting multiple resistance mechanisms involved in Ae. aegypti populations from Bobo-Dioulasso and Ouagadougou (including cytochrome P450). This update to the insecticide resistance status within Ae. aegypti populations in the two biggest cities is important to better plan dengue vectors control in the country and provides valuable information for improving vector control strategies in Burkina Faso, West Africa.

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